With the growth of the global economy, changes in climate and ecological environments, and increased mobility of humans and animals, the transmission risk of vector-borne tropical diseases continues to rise. To address this challenge, strengthening surveillance of vector-borne tropical diseases is urgent. This consensus brought together 29 renowned experts in related professional fields from 26 institutions in China, who, through analyzing the epidemic trend and hazard situation of vector-borne tropical diseases and summarizing the working experiences of experts, have firstly reached following consensus: the burden of vector-borne tropical diseases is heavy with great threats to human health; China has achieved remarkable results in prevention and control of vector-borne tropical diseases, but still needs to strengthen the surveillance and response actively. Secondly, a unanimous consensus has been reached on the aspects of surveillance definition, objectives, contents, and methods of vector-borne tropical diseases. Thirdly, detail requirements have been agreed including: strengthening the concept of early surveillance and forecast, standarding the function, evaluation steps, and construction requirements of surveillance system for vector-borne tropical diseases. Fourthly, key tasks were put forward that need to be investigated and strengthened in the future. This expert consensus provides a standardized reference for the construction of the surveillance pathway and surveillance system for vector-borne tropical diseases in China.

Objective To observe the molluscicidal effect of a new molluscicide, 10% chlorosalicylicamide (LDS), against the intermediate hosts of Schistosoma mansoni - Biomphalaria glabrata and Biomphalaria straminea, thus to provide the experimental foundation for the field application of this molluscicide. Methods The 10% LDS was formulated into the series of standard solutions with effective concentrations of 0.400 0, 0.200 0, 0.100 0, 0.050 0, 0.025 0 mg/L and 0.012 5 mg/L, respectively. B. glabrata and B. straminea were separately immersed in these solutions in the laboratory. The deaths of the above snails were observed after immersing into the solutions for 24, 48, and 72 h, and the mortalities of each group were computed, as well as the median lethal concentration LC₅₀ (s) and relative toxicity indexes were calculated. Meanwhile, 50% wettable power of niclosamide ethanolamine salt (WPN) was set as the drug group, and dechlorinated water as the blank control group. Results The effective concentration of LDS at or above 0.100 0 mg/L, or WPN at or above 0.200 0 mg/L resulted in a 100% mortality rate for B. glabrata and B. straminea after immersing 24, 48, and 72 h. The LC₅₀ (s) at 24, 48, and 72 hours for Biomphalaria glabrata immersed in the LDS series concentration solution was 0.047 95, 0.046 52, and 0.037 10 mg/L, respectively; while the LC₅₀ (s) of B. glabrata in WPN serial solutions was 0.063 48, 0.057 05, 0.057 05 mg/L for 24, 48 and 72 h, respectively. For Biomphalaria straminea, the LC₅₀ (s) at 24, 48, and 72 hours in the LDS solution was 0.012 35, 0.013 99, and 0.008 40 mg/L, respectively; and for the WPN solution, it was 0.058 95, 0.025 71, and 0.0237 5 mg/L. Using WPN as the standard drug which had higher value of LC₅₀, and the relative toxicity index of WPN was set to 1.00, the relative toxicity indexes of LDS against B. glabrata were 1.32, 1.23 and 1.54 for 24, 48 h and 72 h, respectively, while for B. straminea, it was 4.77, 1.84, and 2.83. After 24, 48, and 72 hours of immersion in LDS, the number of surviving B. glabrata was significantly higher than that of B. straminea, with a statistical significance (χ²=8.044, 5.263, 4.658, P<0.05, respectively). Conclusions Compared to the traditional molluscicide WPN, 10% LDS shows a superior molluscicidal effect on B. glabrata and B. straminea, especially demonstrating heightened sensitivity and efficacy on B. straminea, suggesting its potential as a substitute agent for snail control.

Objective To assess the willingness of students with latent tuberculosis infection (LTBI) in Jiangsu Province to undergo preventive treatment and identify factors influencing their decision, aiming to provide insights for tuberculosis prevention and control strategies in school. Methods The physical examination information of tuberculosis latent infection cases was collected from screenings of new school enrollment and contacts of tuberculosis patients in 6 cities of Jiangsu Province from December 2022 to December 2023. Data on past medical history and understanding of preventive treatment were gathered through an online questionnaire survey on the website of Juanxing, and the influencing factors related to the willingness to take preventive medication were analyzed by logistic regression analysis model. Results In December 2022 to December 2023, a total of 13 school tuberculosis outbreaks occurred in 6 cities, and 1 661 contacts were screened, among which 162 cases met the criteria for prophylactic medication, 96 cases were included in the study by filling in the questionnaire. A total of 22 600 new students from 56 schools participated in the TB screening upon enrollment, of which 358 tested positive for the tuberculin skin test alone, meeting the criteria for preventive medication, and 251 of them completed the willingness survey. Finally, 347 students who met the criteria for preventive treatment were included in the study, with 164 expressing to accept preventive treatment representing a treatment acceptance rate of 47.3%. The results of multivariate analysis showed that university (OR=17.950, 95%CI: 3.078-104.686, P=0.001) and contact with the source of school tuberculosis epidemic (OR=19.542, 95%CI: 6.289-60.726, P<0.001) were associated with increased willingness to receive preventive treatment, while unclear whether to pay for the drugs themselves (OR=0.349, 95%CI:0.133-0.916, P=0.032) was associated with decreased willingness to receive preventive treatment. Compared with Huai'an City, the willingness to receive preventive treatment was significantly lower among students from Nantong City (OR=0.005, 95%CI:0.000-0.063, P<0.001), Nanjing City (OR=0.022, 95%CI: 0.003-0.703, P<0.001) and Lianyungang City (OR=0.074, 95%CI:0.008-0.703, P=0.023). Conclusions The acceptance rate of preventive treatment among LTBI students in Jiangsu Province is not high and is affected by multiple factors. Health education and medication mobilization for preventive medication are essential.

Objective To understand the genotyping of human immunodeficiency virus (HIV) co-infected hepatitis C virus (HCV) patients in Yunnan Province, and to analyze the differences in viral load, biochemical indicators, and blood routine indicators among different genotypes, in order to provide a laboratory basis for the diagnosis and clinical treatment of HIV/HCV co-infected patients. Methods From November 2022 to June 2023, the serum samples and basic information of patients diagnosed with HIV/HCV co-infection were collected in the antiviral outpatient clinic of Yunnan Provincial Hospital of Infectious Diseases. The HCV viral load was detected by one-step qRT-PCR amplification, the positive samples were sequenced, and genotyping was determined based on NS5 gene sequence. The differences in biochemical and blood routine indexes between HIV patients co-infected with different HCV genotypes and low/high viral loads were analyzed. Results A total of 126 HIV/HCV co-infected patients were collected, including 20 HCV genotype 1 (15.9%), 91 HCV genotype 3 (72.2%), and 15 HCV genotype 6 (11.9%). The maximum and minimum viral load of the three HCV genotypes were as follows: HCV type 1 (1.0×10⁸, 4.8×10⁴ IU/mL), HCV type 3 (2.2×10⁸, 2.9×10² IU/mL), and HCV type 6 (8.1×10⁷, 6.8×10⁴ IU/mL). The results showed that there was no significant difference between HIV co-infection with different genotypes of HCV and three HIV treatment schemes, including nucleoside reverse transcriptase inhibitors+integrase strand transfer inhibitors (NRTIs+INSTIs), nucleoside reverse transcriptase inhibitors+non-nucleoside reverse transcriptase inhibitors (NRTIs+NNRTIs) and nucleoside reverse transcriptase inhibitors+protease inhibitor (NRTIs+PLs), and the viral load of patients (P>0.05). The analysis of biochemical indexes such as total bilirubin (TBIL), direct bilirubin (DBIL), alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine (CREA), and blood routine indexes such as white blood cell (WBC), red blood cell (RBC), hemoglobin (HGB), platelet (PLT), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) among different HCV genotypes and low/high viral loads showed that there was no significant difference in biochemical indexes and blood routine indexes between low/high viral loads of HIV co-infected HCV patients (P>0.05); however, the biochemical indicators TBIL, IBIL and MCHC were significantly different statistically between patients with genotype 3 HCV infection and those with genotype 1 HCV infection (P<0.05), while other biochemical and blood routine indexes were not statistically different among different HCV genotypes (P>0.05). Conclusions There are six subtypes of HCV co-infection in HIV patients in Kunming, Yunnan Province, including three genes of genotype 1, 3, and 6. Among them, genotype 3 HCV is the main prevalent genetic virus among HIV co-infected populations. The TBIL, IBIL and MCHC values of HIV patients co-infected with HCV type 3 are different from those infected with HCV type 1.

Objective To isolate monoclonal B cells specific for the SARS-CoV-2 receptor binding domain (RBD) from two COVID-19 convalescents, and to screen for broad-spectrum neutralizing antibodies against the SARS-CoV-2 RBD. Methods Using biotinylated RBD as a molecular probe, flow cytometry was employed to perform single-cell sorting of B cells from peripheral blood mononuclear cells (PBMCs) of convalescents. The obtained B cells were lysed and subjected to reverse transcription, followed by nested PCR amplification of the heavy and light chains of antibodies was conducted using random primers. The amplified products were cloned into corresponding expression vectors, and the respective matched heavy-light chain plasmids were co-transfected into 293F cells for expression. Monoclonal antibodies were then purified using Protein A column chromatography. Neutralization experiments were conducted with the wild-type (WT) pseudovirus, and antibodies with IC₅₀<0.1 μg/mL were selected for further testing of neutralizing breadth and potency against the wild-type (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and currently prevalent pseudovirus strains (XBB, BA.5, BF.7). Results A total of 21 RBD-specific monoclonal B cells were obtained from two recovered patients, resulting in the isolation of 13 pairs of antibody light/heavy chains. Nine antibodies were successfully expressed, with P1-A1, P1-B6, and P1-B9 exhibiting IC50 values below 0.1 μg/mL against the pseudovirus of the wild-type strain (WT). Specifically, P1-B6 effectively neutralized the wild-type strain (WT), Beta variant (B.1.351), and Delta variant (B.1.617.2), with IC₅₀ values reaching 0.01 μg/mL. P1-B9 demonstrated effective neutralization against the wild-type strain (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and Gamma variant (P.1) pseudoviruses, with IC₅₀ values of 0.42 μg/mL, 0.63 μg/mL, 0.28 μg/mL, and 2.50 μg/mL, respectively. Additionally, P1-B6 exhibited good neutralization against BA.5 and BF.7 pseudoviruses, with IC₅₀ values of 0.06 μg/mL and 0.09 μg/mL, respectively. Conclusions Infection with the SARS-CoV-2 WT strain can induce the generation of neutralizing antibodies with broad-spectrum activity. Generating these broadly neutralizing antibodies does not require an excessively high somatic hypermutation. The obtained antibodies can be used as candidates for SARS-CoV-2 diagnosis and prevention.

Objective To explore the effect of Mp1p on host macrophages through transcriptomics combined with metabolomics. Methods Firstly, a THP-1 macrophage strain (THP-1-Mp1p⁺) stably expressing Mp1p was constructed using lentivirus. Secondly, using high-throughput RNA sequencing (RNA Seq) technology, the expression level of intracellular mRNA was detected in transcriptomics analysis to determine differentially expressed genes; In metabolomics analysis, metabolite identification was performed through database comparison, and pathway analysis was performed on differential metabolites to reveal potential mechanisms of action. Finally, the results of metabolomics and transcriptomics were combined for analysis, and differential metabolites and genes were analyzed to further elucidate the mechanism of action of Mp1p on macrophages. Results Transcriptome analysis showed that, compared with the negative control group, the THP-1-Mp1p⁺ group had a total of 1 180 differentially expressed genes (DEGs), with 345 upregulated genes and 835 downregulated genes. GO enrichment analysis of DEGs showed that there were 135 differentially expressed genes, including 105 in biological processes (BP), 28 in cellular components (CC), and 2 in molecular functions (MF). The KEGG analysis results showed that the effect of Mp1p on THP-1 macrophages was highly correlated with the TNF pathway. The metabolomic analysis found that both the blank control group and the THP-1-Mp1p⁺ macrophage group achieved good separation between QC samples in both positive and negative ion modes. The threshold for significant differential metabolites was set at: VIP≥1 and T-test P<0.05, resulting in the identification of 488 differential metabolites, with 230 in the positive ion mode and 258 in the negative ion mode. Pathway enrichment analysis of the identified metabolites pointed to significant enrichment in metabolic pathways. The combined analysis confirmed that the tumor necrosis factor signaling pathway, interleukin-17 signaling pathway, and NF-kappaB signaling pathway were important metabolic pathways involved. Conclusions The virulence factor Mp1p may affect host macrophages by modulating the tumor necrosis factor signaling pathway, interleukin-17 signaling pathway, and NF-kappaB signaling pathway. The findings contribute to a better understanding of the mechanisms of action of Mp1p and may offer potential directions for the selection of relevant diagnostic and therapeutic targets in the future.

Objective To evaluate the sensitivity and specificity of the GenoType MTBDRplus V2.0 (Mycobacterium tuberculosis and resistance gene detection assay kit using PCR-linear probe hybridization with enzyme chromogenic method, referred to as MTBDRplus 2.0) kit for detection of rifampin and isoniazid resistance, providing the basis for improving the detection of drug-resistant Mycobacterium tuberculosis. Methods From January to December 2022, positive strains of Mycobacterium tuberculosis isolated and cultured from designated tuberculosis treatment hospitals in 32 counties (cities, districts) of Yunnan Province were collected. Resistance in 880 strains of Mycobacterium tuberculosis was detected by MTBDRplus 2.0, and the minimum inhibitory concentration (MIC) method was used for the drug sensitivity test. Results Using the MIC method as the gold standard, the sensitivity, specificity, positive predictive value, and negative predictive value of MTBDRplus 2.0 in detecting isoniazid resistance were 67.69%, 98.40%, 77.19%, and 97.45%, respectively. The consistency of the linear probe method and MIC method in detecting isoniazid resistance was moderate, with a Kappa value of 0.701 (P<0.001); the sensitivity, specificity, positive predictive value, and negative predictive values of MTBDRplus 2.0 in detecting rifampicin resistance were 87.80%, 99.40%, 87.80%, and 99.40%, respectively. The consistency of rifampicin resistance detection between MTBDRplus 2.0 and MIC method was relatively good, with a Kappa value of 0.872 (P<0.001). Among the 13 strains showing resistance to isoniazid with MTBDRplus 2.0, but sensitivity according to the MIC method, 11 strains (84.62%) had mutations in the inhA gene (C15T). Out of the 5 strains showing resistance to rifampicin with MTBDRplus 2.0, but sensitivity according to the MIC method, 4 strains (80.00%) had other mutations in the rpoB gene. Conclusions MTBDRplus 2.0 shows high sensitivity and specificity in detecting rifampicin resistance, but slightly low sensitivity in detecting isoniazid resistance. The low sensitivity in detecting isoniazid resistance may be due to insufficient target coverage of the detection kit. inhA gene mutations are poorly correlated with isoniazid resistance, and other mutations in the rpoB gene are poorly correlated with rifampicin resistance.

Objective To analyze the clinical characteristics and factors influencing adverse treatment outcomes in elderly patients with multidrug-resistant pulmonary tuberculosis (MDR-TB) to guide the clinical diagnosis and treatment of elderly MDR-TB patients. Methods Clinical data of elderly patients with multidrug-resistant pulmonary tuberculosis initially treated at Beijing Chest Hospital from 2008 to 2023 were retrospectively collected. Complications/comorbidities, adverse drug reactions, drug resistance during initial treatment, and retreatment were statistically described. Factors influencing adverse treatment outcomes were analyzed using the chi-square test and logistic regression analysis. Results A total of 238 elderly patients with MDR-TB were collected, of which 152 (63.9%) had adverse drug reactions, 184 (77.3%) were retreated MDR-TB, 27 (11.3%) were extensively drug-resistant tuberculosis (XDR-TB), 41 were cured, 6 completed treatment, 39 failed treatment, 6 died, 107 lost to follow-up, 31 could not be evaluated, 8 did not finish treatment, and the treatment success rate was 20.4% (47/230). The adverse outcome of treatment accounted for 79.6% (183/230). Univariate analysis showed that differences in age groups, the occurrence of drug adverse reactions, and patient sources had a statistically significant impact on treatment outcomes (P<0.05). Logistic regression analysis was performed using good and adverse treatment outcomes as dependent variables for the three factors, which showed that being aged 70 and above, the occurrence of drug adverse reactions during treatment, and being a non-local patient were factors influencing adverse treatment outcomes [OR (95%CI): 2.507 (1.027-6.121), 3.253 (1.635-6.473), 2.563 (1.285-5.111), respectively]. Conclusions Elderly patients with MDR-TB exhibit a high prevalence of complications/comorbidities, a high incidence of drug adverse reactions, and unfavorable treatment outcomes. Out-of-town medical treatment, advanced age, and experiencing drug adverse reactions are risk factors for adverse treatment outcomes.

Objective To explore the application of time series models based on meteorological factors in the population density of Aedes albopictus in Hainan Province, and to provide a scientific basis for the prevention and control of dengue fever in Hainan Province. Methods The density of Aedes albopictus in different habitats in 18 cities and counties of Hainan Province from 2017 to 2022 was monitored monthly using the double-mosquito net trapping index and the Breteau index. Mann-Kendall trend test was used to analyze the temporal trends of the two density indices; Spearman's correlation analysis was employed to assess the correlation strength between each meteorological factor and the two indexes, eliminating unrelated variables, and further selecting the final variables through the full-subset regression method. Three time-series models were constructed for the two density indices, with root mean square error (RMSE), mean absolute error (MAE), and other accuracy metrics used to determine the optimal model; predictions for the density indices for 2023-2024 were made. All statistical analyses were performed in R (4.3.1). Results The net trapping and Breteau indices showed an overall decrease over the years (Z-values of Mann-Kendall trend test were -6.15 and -4.03, respectively, and P<0.05). The meteorological factor most strongly associated with the trap index was the monthly average minimum temperature; monthly mean minimum temperature and monthly mean relative humidity were strongly correlated with the Breteau index. Based on various evaluation indicators, the multivariate time series model demonstrated the highest accuracy. The study predicts one to two peaks in both the trap index and Breteau Index for the years 2023 and 2024, with peak periods occurring between June to September and May to September, respectively. The predicted value for 2023 aligns with the measured value, demonstrating outstanding predictive accuracy. Conclusions This study has introduced meteorological factors into the seasonal time series model, allowing for more accurate predictions of the density of Aedes albopictus in Hainan Province from 2023 to 2024, providing a model framework for the prevention and control of dengue fever in Hainan Province.

Objective To establish a high-throughput method that can simultaneously, quickly, and accurately detect main malaria-transmitting Anopheles species and their resistance genes in China, providing a high-throughput monitoring tool for monitoring the main malaria vectors in China after malaria elimination. Methods In different sampling locations, including Tengchong City, Yunnan Province; Wenchang City, Hainan Province; and Donggang City, Liaoning Province, adult specimens of mosquitoes, including Anopheles sinensis, Anopheles minimus, Anopheles dirus, and Anopheles anthropophagus, were collected. Polymerase chain reaction (PCR) technology and Sanger sequencing were employed to detect the ITS2, kdr (L1014), rdl (A296), and ace-1 (G119) genes in individual mosquitoes. For the analysis of mixed samples, an optimized multiplex PCR reaction system, custom-designed dual barcode primers, and next-generation sequencing (NGS) technology were utilized to detect the aforementioned genes. The consistency was assessed using Kappa consistency tests and Chi-square tests for multiple rates. Sensitivity, specificity, and the Youden index were calculated using a four-grid table calculation method. The costs associated with each step of the normal operational process for each method were statistically summarized, and the optimal quantity of mixed samples for detection was determined by a comprehensive approach. Results Conventional PCR amplification of gDNA from 300 mosquitoes resulted in 144 individuals of Anopheles sinensis, 53 individuals of Anopheles dirus, 62 individuals of Anopheles anthropophagus, and 41 individuals of Anopheles minimus, as identified by Sanger sequencing. The mutation frequencies of resistance genes kdr (L1014), rdl (A296), and ace-1 (G119) were found in 73, 27, and 41 specimens, respectively. Using a newly established multiplex PCR reaction system based on custom dual barcode and NGS sequencing technology, samples corresponding to Sanger sequencing were detected under different sample sizes. The two methods showed high consistency in the results (all Kappa>0.900). Multiple comparison tests showed significant differences in the consistencies of the two methods across different sample sizes N (40, 80, 160), N (120, 200, 240, 280), and N (300) (χ²=26.547, P<0.001). The new method demonstrated high sensitivity and specificity across various sample sizes, with the Youden index ranging from highest to lowest as follows: 1 (40, 80, 160)>0.994 (120)>0.990 (280)>0.988 (200)>0.987 (240)>0.985 (300). With an increase in sample size from 40 to 300, the cost per sequencing site for the new method decreased from 20.0 yuan to 8.3 yuan, while the cost per sequencing site for the conventional method decreased from 16.7 yuan to 15.4 yuan. The optimal mixed sample size for the new method was determined to be 280. Conclusion The newly developed multiplex PCR and barcode NGS detection method enables simultaneous screening of four major malaria vector mosquito species and the presence of mutations in the ace-1, kdr, and rdl resistance genes, exhibiting excellent stability, high sensitivity, and specificity. It allows for the efficient analysis of large sample sizes in a single run, offering a cost-effective alternative compared to other types of detection methods.

Objective To investigate the resistance level of Aedes albopictus to commonly used insecticides in Jinshan District, Shanghai, to provide a reference for standardizing the use of insecticides. Methods The larval dipping test was used to detect the resistance of Ae. Albopictus larvae to five kinds of insecticides, the SPSS 18.0 software was utilized to calculate the toxic regression equation and the median lethal concentration (LC₅₀) of insecticides on the larvae. The resistance level was determined by evaluating the 24-hour mortality of adult Ae. Albopictus exposed to diagnostic doses of commonly used insecticides with the adult mosquito contact tube method. Results In 2018 and 2019, Ae. Albopictus larvae in the Jinshan District of Shanghai displayed moderate and high resistance to beta-cypermethrin, with resistance ratios of 25.03 and 65.96 folds respectively; high resistance to deltamethrin, with resistance ratios of 57.25 and 211.75 folds respectively; high resistance to permethrin, with resistance ratios of 46.17 and 243.36 folds. In 2018, 2019, 2021, and 2023, they showed moderate to high resistance to temephos with resistance ratios of 19.55, 23.94, 53.48, and 22.12 folds respectively. In 2021 and 2023, moderate resistance to fenitrothion was observed, with resistance ratios of 30.04 and 12.54 folds respectively. Adult Ae. Albopictus adults exhibited resistance to 0.03% deltamethrin, 0.07% lambda-cyhalothrin, 0.4% permethrin, and 0.08% beta-cypermethrin, with mortality rates ranging from 17.20% to 49.67% in 2021 and 2023. Potential resistance was observed to 0.7% lambda-cyhalothrin and 0.2% fenitrothion, with mortalities of 97.48% and 83.74% respectively. Sensitivity was noted to 0.05% propoxur with a mortality rate of 100.00%. Conclusions Ae. Albopictus in the Jinshan District, Shanghai, has developed varying resistance levels to different types of insecticides, including pyrethroids and organophosphates. It is recommended to strengthen the dynamic monitoring of the resistance of Ae. Albopictus and implement comprehensive prevention and control measures with a focus on environmental management, scientifically and rationally selecting hygienic insecticides to delay and mitigate the emergence of resistance.

Objective To investigate whether the expression regulation of regulatory gene Sigma factors (sigA-sigM) is related to isoniazid resistance phenotype in isoniazid-resistant Mycobacterium tuberculosis (MTB) caused by katG mutations, and to provide reference for the study of the molecular mechanism of isoniazid resistance. Methods A total of 90 strains were collected from the patients undergoing first-line treatment at the Tianjin Tuberculosis Control Center during drug resistance testing from 2020 to 2022, of which 30 strains were sensitive strains without katG mutation, and 65 strains were isoniazid-resistant strains caused by katG mutation, including 11 strains resistant only to isoniazid, 24 strains resistant to both isoniazid and streptomycin, and 30 strains multidrug-resistant to isoniazid, streptomycin, and rifampicin. After the strains were collected on the isoniazid drug-containing medium, the RNA of the strains was extracted, and the relative expression levels of Sigma factors were detected by reverse transcription-polymerase chain reaction(RT-PCR). The expression differences of Sigma factors in different isoniazid drug-resistant phenotypes were analyzed by the Mann-Whitney U test. Results The gene expression levels of sigA, sigC, sigF, sigG, sigH, sigI, sigJ, sigK, sigL in isoniazid mono-resistant group were significantly higher than those in pan-isoniazid-sensitive group (Z=4.368, 5.701, 6.865, 4.021, 5.126, 2.670, 5.983, 4.701, 5.490, P all<0.001). The gene expression levels of sigA, sigC, sigF, sigG, sigH, sigI, sigJ, sigK, sigL in poly-resistance group were significantly higher than those in pan-sensitivity group (Z=-5.017, -4.670, -4.667, -5.456, -4.083, -5.393, -4.712, -6.971, -8.206, -5.211, P all<0.001). In multidrug-resistant (MDR) group, the gene expression levels of sigC, sigD, sigE, sigF, sigG, sigH, sigI, sigJ, sigK, sigL were significantly higher than those in pan-isoniazid-sensitive group (Z=-5.537, -4.003, -5.216, -7.328, -7.730, -5.658, -4.440, -6.036, -4.862, -4.312, P all<0.001). The expression levels of sigB, sigF, sigG showed statistically significant differences in gene expression between the isoniazid mono-resistant, isoniazid poly-resistant, and isoniazid multidrug-resistant groups (Z=10.139, 7.735, 14.532, P all<0.001). The expression rates of sigF, sigG, sigI, sigJ, and sigL in the isoniazid-resistant group were significantly higher than those in isoniazid sensitive group (χ²=17.410. 45.673. 57.661. 42.896. 26.363, P all<0.001). Conclusions sigF, sigG, sigI, sigJ, and sigL are associated with isoniazid resistance due to katG mutations.

Objective To investigate the current status and attrition among HIV-infected persons receiving antiretroviral therapy (ART), and to analyze factors affecting attrition in Hainan. Methods In this study, HIV-infected patients who started ART treatment in Hainan Province from 2005 to 2022 were selected from the antiviral treatment submodule of China Disease Prevention and Control Information System.According to the inclusion and exclusion criteria,a total of 4 286 HIV-infected persons were receiving. A Cox proportional hazards regression model was used to analyze factors affecting attrition. Results Among the 4 286 study subjects, 3 718 were males (86.7%), with a sex ratio of 6.55∶1. Unmarried individuals accounted for 58.4%, and the average age was (39.68±13.17) years. Transmission through homosexual contact accounted for 49.8%, and 84.3% were in WHO clinical stage I. Treatment regimens containing Efavirenz (EFV) accounted for 71.7%. During a follow-up of 19 677.44 person-years, the overall attrition rate was 0.80 per 100 person-years, with the first-year post-ART initiation attrition rate being 21.10 per 100 person-years. The results of Cox regression analysis showed that the time of treatment initiation in 2016-2022 (AHR=2.40, 95%CI: 1.40-4.10), and the last HIV viral load (VL) 20-<1 000 copies/mL (AHR=3.69, 95%CI: 2.08-6.54), the last HIV-1 VL≥1 000 copies/mL (AHR=15.98, 95%CI: 9.46-27.01), and no last HIV-1 VL test (AHR=92.90, 95%CI: 57.68-149.62), the time interval from diagnosis to treatment for 1-12 months (AHR=1.62, 95%CI: 1.12-2.36), and an interval longer than 12 months (AHR=1.68, 95%CI: 1.07-2.62) were the main factors that increased the risk of attrition. Treatment regimens containing Lopinavir/ritonavir (Lpv/r) (AHR=0.34, 95%CI:0.18-0.66) and treatment regimens containing integrase strand transfer inhibitors (INSTIs) (AHR=0.24, 95%CI: 0.09-0.58) were the factors that reduced the risk of attrition after antiretroviral therapy. Conclusions The attrition of ART in HIV/AIDS patients in Hainan Province is related to a longer interval from diagnosis to treatment, treatment plan, and abnormal HIV viral load test results. Case-based measures should be taken to address factors influencing antiretroviral treatment attrition, while improving the timeliness of antiviral treatment and treatment management service quality to further improve the efficacy of antiviral treatment.

Objective To perform the pathogenic and genomics analyses on isolates of Streptococcus suis (Ss) from three human infections in Shenzhen, aiming to provide a basis for the prevention and control of Ss outbreaks. Methods The suspected bacterial strains from three blood plate cultures of three critically ill patients in three hospitals were subjected to biochemical identification, matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), and real-time fluorescent PCR identification, resulting in the identification of three strains positive for Streptococcus suis serotype 2(SS2). Pure positive cultures were taken for an antimicrobial susceptibility test and extracted nucleic acids for whole-genome sequencing and analysis. The whole-genome sequencing and analysis included species identification, antibiotic resistance genes alignment, multilocus sequence typing (MLST), virulence genes alignment, and coregene-based phylogenetic tree analysis. Results The blood agar isolates from three patients were all identified as Ss, the VITEK 2 identified them as SS2, and MALDI-TOF-MS identified them as Ss. Real-time PCR results for the universal gene gdh and serotype 2 cps2 gene of Ss were both positive. The antimicrobial susceptibility test results showed that all three strains were resistant to erythromycin and clindamycin, with variable sensitivity to tetracycline. Whole-genome sequencing results showed that all three strains were identified as Ss, including one ST7 strain and two ST1 strains. The virulence gene prediction results based on the VFDB database showed that all three strains were positive for mrp, sly, and cps, indicating high virulence gene characteristics. The analysis of the phylogenetic tree based on coregene showed that the three strains were in different evolutionary branches, with two ST1 strains having a closer evolutionary distance. Conclusions The pathogens responsible for these three critically ill patients were SS2, and all three strains were resistant to erythromycin and clindamycin. Genetically, they all carried virulence genes that are found in highly virulent strains, while showed differences in MLST typing and phylogenetic tree analysis, indicating the presence of different genotypes of high pathogenicity SS2 in Shenzhen area and had caused sporadic cases, which requires high attention.

Objective To conduct preliminary research on the prevalence and genotyping of urogenital Chlamydia trachomatis (CT) infection in sexually transmitted disease (STD) clinics in Hainan Province, to understand the epidemiological characteristics and genotype features of the population infected with urogenital CT, and to provide evidence for the formulation of scientific prevention strategies and measures. Methods From 2018 to 2022, a total of 5 551 male urethral swabs and female cervical swabs were collected for detection of CT infection by real-time fluorescent quantitative polymerase chain reaction (RT-PCR). The OMPL gene was amplified from the DNA of some CT positive individuals by nested PCR, and the positive results were sequenced. Sequencing results were uploaded to BLAST website to find sequence similarity and construct a phylogenetic tree to determine the genotype. Results Out of the 5 551 tested patients, 846 were positive for Chlamydia trachomatis infection, with a positivity rate of 15.2%, the positive detection rate of CT-DNA was 18.6% in male and 13.4% in female, the positive detection rate of male was higher than that of female. There were statistically significant differences in the CT-DNA positivite detection rate among different age groups (P<0.05), and the highest positivite rate CT-DNA was 58.0% in 20-<30 years old, while it was the lowest, at 1.0%, in those over 50; there were also significant differences in CT-DNA positivity detection rate between seasons (P<0.05), with the highest being 36.4% in the summer and the lowest at 9.6% in the winter. Genotyping of the CT-OMPL gene VS1-VS2 nucleotide sequence in some samples from the Hainan region identified six genotypes, including types D, E, G, F, J, and K, with type F being the main prevalent dominant type. Conclusions CT infection in Hainan is associated with gender, age, and season, and the genotypes are diversified. It is necessary to further strengthen the screening of CT infection in the reproductive tract of men and women of childbearing age in the future STD prevention and treatment work to improve the fertility rate.

Objective To investigate the differential expression of proteins in the serum of diabetic patients quantitative proteomics technologies, and to identify specific protein biomarkers. Methods Serum samples from three diabetic patients and three healthy controls in Shenzhen in 2023 were collected. Immunoprecipitation and nano liquid chromatography-quadrupole-electrostatic orbitrap-linear ion trap mass spectrometry (easy-nanoLC-Q-orbitrap-fusion) were employed to perform fraction and no-fraction-based quantitative analysis of the serum proteome in diabetic patients and healthy controls. The mass spectrometry data were analyzed by MaxQuant1.6.1.0, and the differential proteins identified by the fraction-based method were subjected to gene ontology (GO) functional clustering and kyoto encyclopedia of genes and genomes (KEGG) pathway analysis. Results A total of 251 reliable proteins were identified by the no-fraction-based method, among which 17 differentially expressed proteins were selected. In contrast, 494 reliable proteins were identified by the fraction-based method, and 74 differentially expressed proteins were selected. Among them, seven proteins showed consistent changes in both methods. Five proteins were downregulated, including pregnancy zone protein (PZP), carbonic anhydrase 1 (CAH1), mannose-binding protein C (MBL2), fibrinogen alpha chain (FIBA), and haptoglobin (HPT). Two proteins were upregulated, including apolipoprotein(a) (LPA) and complement factor H-related protein 4 (CFH4). GO analysis revealed that the differentially expressed proteins were mainly involved in molecular functions such as glycosaminoglycan binding and heparin binding. KEGG pathway analysis showed that the differentially expressed proteins were mainly associated with complement and coagulation cascades, cell adhesion, PI3K-Akt signaling pathway, and other pathways. Conclusions Fraction-based no-fractional quantitative proteomics analysis provides an effective approach for analyzing differential protein expression in the serum of diabetic patients, and offers a valuable technique for screening specific protein biomarkers of serum samples in large-scale cohorts of disease.

Objective To investigate the relationship between exosomal microRNA (miR)-335-5p and the severity of cirrhosis in patients with chronic hepatitis B (CHB), in order to provide more clinical information for early intervention and treatment. Methods From February to August 2019, 6 healthy controls, 8 cases of compensated cirrhosis (CLC), and 8 cases of decompensated cirrhosis (DLC) were recruited from the physical examination outpatient department and the hepatology department of Peking University Third Hospital Qinhuangdao Hospital as a discovery cohort, and serum samples were collected for exosome extraction and miRNA microarray analysis. A validation cohort of 229 CHB patients with cirrhosis, including 94 CLC and 135 DLC patients, was selected from those diagnosed and treated in the hepatology department of the hospital from December 2019 to May 2022. Exosomes were extracted and identified, and the expression level of serum exosomal miR-335-5p was detected by real-time fluorescence quantitative PCR. The model for end-stage liver disease (MELD) score and the combined formula of serum sodium with MELD (MELD-Na) score were used to evaluate the severity of cirrhosis. Results In the discovery cohort, the expression of miR-335-5p in the DLC group was significantly down-regulated compared to the control group and CLC group (P<0.05). In the validation cohort, the level of exosomal miR-335-5p in the DLC group was significantly lower than that in the CLC group (P<0.001). Spearman correlation analysis showed a negative correlation between the expression of exosomal miR-335-5p and both MELD and MELD-Na scores in patients with cirrhosis (P<0.05). After adjusting for other confounding factors in the multiple linear regression model, the expression of exosomal miR-335-5p was still negatively correlated with MELD score (β=-0.103, 95%CI:-3.692 to -1.149, P<0.001) and MELD-NA score (β=-0.109, 95%CI:-4.007 to -1.270, P<0.001). ROC curve analysis indicated that serum exosomal miR-335-5p could differentiate DLC with an area under the ROC curve of 0.905 (95%CI: 0.867 to 0.944), corresponding to a cutoff value of 0.158, with a specificity of 87.2%, and a sensitivity of 83.7%. Conclusions The low expression of exosomal miR-335-5p is associated with the aggravation of cirrhosis in CHB patients, and may serve as a useful biomarker for the early diagnosis of DLC.

Objective To analyze the current situation and influencing factors of health-care seeking delay among pulmonary tuberculosis patients in Qingpu District of Shanghai from 2011 to 2022, and to provide a scientific basis for tuberculosis prevention and control. Methods The data of pulmonary tuberculosis patients in Qingpu District of Shanghai from 2011 to 2022 was collected through the China Tuberculosis Information Management System to describe the distribution and change trend of the delay in health-care seeking. Univariate analysiswas performed using the chi-square (χ²) test, and the time trend of rates was tested with the trend chi-square (trend χ²) test. Multivariate logistic regression model analyzed the influencing factors of the delay in health-care seeking. Results From 2011 to 2022, there were 3 488 cases of pulmonary tuberculosis in Qingpu District, with 1 438 patients experiencing health-care seeking delay. The median (quartile) number of days of delay was M (P₂₅, P₇₅) = 10 (2, 24) days, and the rate of health-care seeking delay was 41.23%. The annual rate of health-care seeking delay fluctuated between 33.88% and 50.45% from 2011 to 2022, with statistically significant differences between different years (χ²=38.355, P<0.001), and an upward trend in the health-care seeking delay rate was observed from 2020 to 2022 (χ_trend²=13.290, P<0.001). Multivariate logistic regression analysis showed that compared to male, those under 25 years old, with local household registration, and detected through health check-ups, females (OR=1.21, 95%CI:1.04-1.41), those aged 45 to <65 (OR=1.36, 95%CI:1.06-1.75), intra-city migrants (OR=1.35, 95%CI:1.09-1.68), inter-provincial/overseas migrants (OR=1.50, 95%CI:1.23-1.83), and patients who directly sought medical care (OR=3.52, 95%CI:2.27-5.47), transfer treatment (OR=2.07,95%CI：1.31-3.25), referral (OR=2.16, 95%CI:1.36-3.44), follow-up (OR=3.07, 95%CI:1.74-5.44) patients with pulmonary tuberculosis were more likely to delay health-care, and the differences were statistically significant (P<0.05). Compared to sputum-positive patients, those with sputum-negative tests (OR=0.76, 95%CI: 0.59-0.97) were less likely to experience delayed health-care, and the difference was statistically significant (P<0.05). Conclusions Health-care seeking delay of pulmonary tuberculosis patients is relatively common in Qingpu District of Shanghai. Corresponding intervention measures should be adopted for risk factors and key populations to further improve the health-care seeking delay.

Objective To establish a dual droplet digital PCR (ddPCR) assay for herpes simplex virus type I (HSV-1) and varicella-zoster virus (VZV). Methods The specific primers and probes were derived based on the conserved regions of HSV-1 and VZV genome. The primer-probe combinations were screened, and the annealing temperatures and primer-probe concentration ratios of the dual-droplet digital PCR reaction were optimized to establish a dual-droplet digital PCR reaction system for HSV-1 and VZV, which was tested for other viruses and validated for clinical samples. The sensitivity, specificity, and reproducibility of the established dual microtiter digital PCR method were analyzed. Results The optimal concentrations of primers and probes for the dual ddPCR detection method of HSV-I and VZV were determined to be 800 nmol/L and 250 nmol/L, respectively, with an optimal annealing temperature of 56 ℃. The correlation coefficient (R²) of the standard curve of the dual ddPCR assay was 0.99, showing a clear linear relationship. The method showed high sensitivity, with the lowest detection limit of herpes simplex virus type I being 2.97 copies/μL, and for VZV being 2.73 copies/μL. The repeatability was high with a small coefficient of variation and stable detection results; the specificity was excellent, and no cross-reaction was found with herpes simplex virus type Ⅱ, Epstein-Barr virus, Adenovirus, Coxsackievirus (CA6/CA10/CA16), Cytomegalovirus, Human Cytomegalovirus, Human enterovirus 71, Japanese Encephalitis virus, West Nile virus, Measles virus, Mumps virus, and human nucleic acids. Conclusions The dual droplet digital PCR assay for herpes simplex virus type I and varicella-zoster virus established in this experiment has strong sensitivity, specificity, and high repeatability, and can provide a solution for rapid quantitative detection of the two viruses in different scenarios.

A confirmed case of blood flow infection caused by Vibrio vulnificus at Fenyang Hospital in Shanxi Province is reported to provide a reference for the diagnosis and treatment of Vibrio vulnificus infection. The patient is a male sanitation worker who accidentally scratched his right forearm while working and was admitted to the hospital due to swelling and pain in his right forearm accompanied by fever. The patient had recently resided in the local area, and had no history of working outside, or recent exposure to seawater or raw seafood. On the morning of the second day of admission, all four blood culture bottles tested positive. The positive blood culture bottle broth was directly smeared and transferred to a blood plate or a hemophilic chocolate plate. Under the microscope, it was Gram-negative bacilli, and the bacteria body was curved and rod-shaped. In the afternoon (about 7 hours of cultivation), the bacteria membrane was identified as Vibrio vulnificus by matrix-assisted laser desorption ionization-time of flight mass spectrometer (MALDI-TOF MS). Sepsis caused by Vibrio vulnificus has a high mortality rate, and it can also cause necrotizing fasciitis. With the cooperation of multiple disciplines, the patient improved and was discharged. Through this case, Vibrio vulnificus infection should be included in the differential diagnosis for patients with fever, redness, swelling, and pain in the limbs (with a history of trauma). The patient's prognosis can be improved with the timely finding of the pathogenic bacteria through blood culture and other techniques, early surgical intervention and establishment of an MDT (multidisciplinary joint) team.

Vibrio vulnificus infection has a high disability rate and fatality rate. This paper summarized the diagnosis and treatment of a case of Vibrio vulnificus sepsis to help clinicians in the early detection and optimization of treatment options for such infections, thereby providing a clinical reference for related research. In September 2022, a case of sepsis caused by Vibrio vulnificus infection was admitted to the Emergency Department of Hainan Hospital of Traditional Chinese Medicine. Clinical treatment was conducted, and the report was analyzed. Meanwhile, Vibrio vulnificus literature was reviewed to analyze and summarize the treatment methods of Vibrio vulnificus sepsis. The 61-year-old patient was treated in the Emergency Department from September 23 to October 8, 2022, during which bacteriological examination of pus and wound infection samples identified the infection as being caused by Vibrio vulnificus. Therefore, targeted treatment with meropenem injection plus levofloxacin injection was administered for the infection, and early CRRT (continuous renal replacement therapy) was implemented to control the inflammatory cytokine storm of sepsis, and surgical debridement treatment was performed. After the patient's condition stabilized, the patient was transferred to the vascular surgery department, where two debridement + left lower limb skin grafting procedures were performed on October 27 and November 19. Then a third debridement + left lower limb third skin grafting + toe amputation was performed on December 10, the patient was discharged with improved condition post-surgery. At present, the patient can live independently and can move the affected limb autonomously. In clinical diagnosis and treatment of sepsis caused by Vibrio vulnificus infection, attention should be paid to the inflammatory cascade reaction. CRRT can eliminate inflammatory factors in the early stage, sensitive antibiotics for anti-inflammatory, surgical debridement interventional treatment, and multi-disciplinary linkage can be initiated to treat patients with Vibrio vulnificus sepsis.

To analyze a case of severe avian influenza A (H5N6) virus infection resulting in severe pneumonia and acute respiratory distress syndrome (ARDS) was admitted to Guilin Municipal Hospital of Traditional Chinese Medicine on July 6, 2023. The clinical data and treatment of this patient were analyzed retrospectively. The initial clinical manifestations of the patient were fever, cough, and expectoration, and the antigen test for influenza A virus was positive. Chest CT showed: double lung texture increased and thickened, and multiple patchy high-density shadows with air-containing bronchial shadows were found in the left lung, especially in the left upper lobe; a few patchy increased-density shadows were also seen in the lower lobe of the right lung, along with left-sided pleural effusion. Metagenomic next-generation metagenomic sequencing (mNGS) of bronchoalveolar lavage fluid was performed to identify the pathogen as influenza A virus H5N6. On the 4th day of admission, the patient's condition rapidly progressed to ARDS, which could not be improved by high-flow oxygen therapy, mechanical ventilation, and prone position ventilation. Subsequently, with the assistance of veno-venous extracorporeal membrane oxygenation (VV-ECMO), the patient's lung function gradually improved. Extracorporeal membrane oxygenation（ECMO） was withdrawn after 25 days, and the patient recovered and was discharged after a hospital stay of 41 days. Patients with severe avian influenza A (H5N6) usually have critical illness and rapid progression, often rapidly progressing to ARDS. When conventional mechanical ventilation cannot correct hypoxemia, VV-ECMO auxiliary treatment should be administered as early as possible. In addition, mNGS can help to quickly identify the diagnosis and differential diagnosis of avian influenza A (H5N6) in the early stage of the disease, particularly suitable for the diagnosis of severe and emergency infections.

This article reported the diagnosis and treatment of a case of fascioliasis as well as an epidemiological investigation conducted in Laibin City, Guangxi, China. The patient came to the hospital for bellyache. Based on the patient's main symptoms, physical signs, clinical presentation, and laboratory examination (abdominal ultrasound scan and CT), fasciola eggs were detected through the examination of stool specimens using the sedimentation method. Fasciola egg DNA was extracted, and after amplification, sequencing, and comparison of the ITS2 gene, a homology of 99.2% with Fasciola gigantica ITS2 sequences in the NCBI database confirmed the diagnosis of Fasciola gigantica infection. After triclabendazole treatment, the patient's symptoms disappeared, and stool examination yielded negative results for Fasciola eggs. A survey of 33 human fecal samples from the patient's vicinity all tested negative, while examination of 10 samples of water buffalo feces from around the residence found Fasciola eggs in eight samples (80%), confirmed as Fasciola gigantica through ITS2 gene amplification and sequencing; microscopic examination of 200 specimens of Galba pervia and 20 physaacuta revealed no positive findings; Additionally, observation of 10 samples of pickled Colocasia gigantea (a local edible plant) that the patient had cultivated and consumed raw revealed no Fasciola metacercariae attachment. The results indicate that this case was an accidental case of Fasciola infection. Due to the lack of specificity in clinical symptoms, it is necessary to combine stool examination for Fasciola eggs and molecular diagnosis to confirm the condition.