Chronic hepatitis B (CHB) caused by the hepatitis B virus (HBV), constitutes a significant public health issue, with approximately 254 million individuals affected globally, of which 75 million are in China. HBV infection not only severely impacts individual health, leading to conditions such as liver cirrhosis, hepatocellular carcinoma, and liver failure, but also imposes considerable economic burdens on society. This review provides an overview of the current epidemiological characteristics and disease burden of CHB, along with the progress towards, and challenges faced in achieving the World Health Organization's (WHO) goal of eliminating viral hepatitis as a public health threat by 2030. It delves into strategies aimed at reducing new infections, enhancing diagnosis rates, and improving treatment coverage. These strategies include universal screening, expanding the scope of antiviral therapy, and implementing specific measures for treating patients with end-stage liver disease. The review underscores critical actions necessary to accelerate the elimination of hepatitis B-related harms, proposing an integrated control and prevention strategy that incorporates new technologies and approaches. Achieving WHO's 2030 target requires concerted efforts from governments, healthcare institutions, and the broader community, alongside active public participation and support.
Objective To screen genetic susceptibility markers related to serum HBeAg seroconversion by analyzing the association between host genetic susceptibility markers and seroconversion in HBeAg-positive patients with chronic hepatitis B (CHB), thereby providing potential molecular markers for clinical outcomes and prognosis evaluation in HBeAg-positive patients with CHB. Methods HBeAg-positive patients with CHB were recruited from the outpatient department of the Infectious Department, Second Affiliated Hospital of Air Force Military Medical University to establish a follow-up cohort. Based on whether HBeAg seroconversion occurred during follow-up, the subjects were divided into a case group (serum HBeAg with seroconversion) and a control group (serum HBeAg without seroconversion). MassARRAY SNP genotyping technique was used to detect SNPs of human genome DNA extracted from whole blood of the study subjects. Results Through association analysis between genetic susceptibility marker SNPs and seroconversion of serum HBeAg, it was found that: At the rs101206 8 locus, under the overdominant model, patients carrying the heterozygous GT genotype had a higher likelihood of serum HBeAg seroconversion (OR=1.73, 95%CI: 1.15-2.59, P=0.008); at the rs352140 locus, under the recessive model, patients carrying the TT genotype had a higher likelihood of serum HBeAg seroconversion (OR=1.77, 95%CI: 1.06-2.94, P=0.029); the rs1946518 locus, under the overdominant model, patients carrying the heterozygous GT genotype had a higher likelihood of serum HBeAg seroconversion (OR=1.73, 95%CI: 1.14-2.61, P=0.009); at the rs2306494 locus, under the dominant model, carrying the A allele (GA+AA) was identified as a negative factor for serum HBeAg seroconversion (OR=0.65, 95%CI: 0.42-0.99, P=0.043); at the rs20541 locus, under the recessive model, carrying the AA genotype was identified as a negative factor for serum HBeAg seroconversion (OR=0.31, 95%CI: 0.10-0.92, P=0.019); at the rs1057035 locus, under the dominant model, patients carrying the C allele (CT+CC) had a higher likelihood of serum HBeAg seroconversion (OR=1.78, 95%CI: 1.05-3.03, P=0.034). Therefore, the GT genotype at rs1012068 of DEPDC5 gene, TT genotype at rs352140 of TLR9 gene, GT genotype at rs1946518 of IL18 gene, and GG genotype at rs2306494 of TERF1 gene were conducive to seroconversion of serum HBeAg. The AA genotype at rs20541 of IL-13 gene and the TT genotype at rs1057035 of DICER1 gene were not conducive to seroconversion of serum HBeAg. Conclusion The genetic susceptibility markers (single nucleotide polymorphism loci) of host genetic genes in HBeAg-positive patients with CHB are associated with seroconversion of serum HBeAg, and the mechanisms by which these SNPs participate in serum HBeAg seroconversion require further investigation.
Objective To monitor the dynamic changes in liver stiffness and their correlation with clinical indicators among chronic hepatitis B virus (HBV) infected individuals in Wuwei City, Gansu Province, and to provide important evidence for the early detection and effective intervention of liver fibrosis (LF) progression. Methods Based on the Wuwei Hepatitis B Specialty Cohort, this study conducted annual serological and FibroScan ultrasonographic examinations for 3 882 chronic HBV-infected individuals. Over an average of 4 years of follow-up, the liver fibrosis outcome was monitored, and influencing factors were analyzed by constructing a logistic regression model. Results Among the 2 053 chronic HBV-infected individuals who completed at least one follow-up, baseline LF grades F0 to F4 were distributed as follows: 1 581 cases (77.0%), 164 cases (8.0%), 99 cases (4.8%), 110 cases (5.4%), and 99 cases (4.8%), respectively. Significant differences were observed among the five groups in terms of age, gender, smoking, antiviral treatment, liver function indicators, control attenuation index (CAP), and liver stiffness measurement (LSM) (P<0.05). After an average of 4 years of follow-up, 1 686 cases (17.9/100 person-years) showed no significant change in LF grade, 260 cases (2.8/100 person-years) demonstrated a decrease in LF grade, and 107 cases (1.1/100 person-years) exhibited an increase in LF grade. Stratified by baseline treatment status, among patients with chronic HBV infection who did not undergo treatment, baseline alanine aminotransferase (ALT) (OR=5.50, 95%CI：1.79-16.83, P=0.003) and LSM (OR=3.35, 95%CI：1.23-9.13, P=0.018) were identified as risk factors for LF progression. In contrast, among patients who underwent antiviral treatment, baseline aspartate aminotransferase (AST) (OR=2.23, 95%CI：1.41-3.53, P<0.001) and total bilirubin (TBIL) (OR=1.79, 95%CI：1.14-2.81, P=0.012) levels were identified as risk factors for LF progression. Conclusion LSM and liver function indicators, such as ALT, AST, and TBIL, are important influencing factors for LF progression. The monitoring of LSM and liver function indicators will be of great significance for the prevention and early diagnosis of liver cirrhosis.
Objective To analyze the hepatitis B (HBV) incidence trend and intervention measures implemented over 15 years (2009-2023) following the establishment of Wuwei demonstration zone and to explore a new model suitable for hepatitis B prevention and control in rural areas of western China. Methods Data on reported hepatitis B cases (1994-2023) were collected from Wuwei City's Disease Monitoring Report Management Information System, vaccination records from the Immunization Planning Information Management System, hepatitis B maternal-to-child transmission prevention data from the Management Information System for Prevention of HIV, Syphilis, and Hepatitis B Mother-to-Child Transmission. Additionally, field epidemiological survey data, serological test results, and intervention measures were also used to analyze using descriptive epidemiological methods. Results Over the 15-year period, the average annual incidence rate of hepatitis B in the demonstration area was 200.48 per 100 000, representing a 38.76% reduction compared to the pre-construction period (1994-2008, 327.35 per 100 000). Post-establishment, the incidence decreased from 637.70 per 100 000 in 2009 to 87.45 per 100 000 in 2023, showing an average annual decline of 13.23%. In intensified intervention areas, the incidence decreased by 89.28%, and the rate stabilized in the mid-to-late stages. Among pregnant women, the HBsAg-positive rate was 4.41%, with 99.42% of newborns born to HBsAg-positive mothers receiving hepatitis B immunoglobulin immunization. The prevalence of chronic hepatitis B, liver cirrhosis, and liver cancer-related diseases decreased by 42.19%, while the mortality rate of hepatitis B patients declined from 8.69% to 2.00%. Public awareness of hepatitis B prevention and control increased from 43.48% to 83.79%. A hepatitis B prevention and control model suitable for impoverished western regions—the “Wuwei Model”—was established and validated. This model emphasizes short-course vaccination, early intervention, integration of prevention and treatment, and collaborative efforts. Conclusion The “Wuwei Model” significantly reduced the incidence and mortality of hepatitis B in the demonstration area, demonstrating effective disease control. Further validation and promotion of this model are needed. Continued efforts should focus on expanding whole-population screening for chronic hepatitis B infection, improving the management of chronic hepatitis B carriers, and increasing immunization rates among susceptible adult populations.
Objective To investigate the association between neutrophil-to-lymphocyte ratio (NLR) and the state of sustained immune response after hepatitis B vaccination, to explore the possibility to predict the effect of hepatitis B vaccination. Methods Data were analysed from 10 153 individuals aged 18 years and older who had received three doses of hepatitis B vaccine and met the inclusion criteria in the National Health and Nutrition Examination Survey (NHANES) 1999-2018. They were divided into seropositive and seronegative groups based on the results of post-vaccination hepatitis B marker testing. The association between NLR and sustained immune response status after hepatitis B vaccination was assessed using a variety of methods, including multivariate weighted logistic regression models, generalised linear analysis, and smoothed curve fitting. Results This study included 10 153 subjects with a mean age of 35.36 years, 42.82% male and 57.18% female. Adjusted multivariate weighted logistic regression models showed that for each unit increase in NLR level, the odds of a sustained immune response after hepatitis B vaccination decreased by 7% (OR=0.93, 95%CI: 0.88-0.97). Stratified analysis of NLR levels showed that the odds of a sustained immune response were 37% lower in the group with the highest NLR levels (T4) compared with the lowest group (T1) (OR=0.63, 95%CI: 0.47-0.85), and the odds of a sustained immune response were 30% lower in the T3 group compared with the T1 group (OR=0.70, 95%CI: 0.53-0.91). Restricted cubic spline analysis showed a non-linear relationship between NLR and sustained immune response status after hepatitis B vaccination (P non-linear<0.05), approximating an 'L' curve. Inflection point analysis showed that when NLR <2.16, elevated NLR significantly reduced the occurrence of sustained immune response status after hepatitis B vaccination; when NLR >2.16, the effect of its elevation on the production of sustained immune response after vaccination leveled off. Conclusion NLR levels were negatively associated with the development of a sustained immune response after hepatitis B vaccination. Lower NLR levels have a strong positive correlation with sustained immune response after hepatitis B vaccination, whereas higher NLR levels may attenuate the effect of hepatitis B vaccination.
Objective This study aims to investigate and compare the clinical characteristics and prognostic factors among primary liver cancer (PLC) patients who had hepatitis B virus (HBV)-induced cirrhosis associated with liver cancer, alcoholic cirrhosis associated with liver cancer, or both HBV and alcoholic cirrhosis associated with liver cancer. Methods Inpatients diagnosed with PLC admitted to the First Affiliated Hospital of Fujian Medical University between January 2010 and September 2020 were enrolled and divided into three groups based on the etiology. The follow-up period ends in October 2024. Survival analyses were performed using Kaplan-Meier curves, univariate analysis, and multivariate Cox regression. Results During the study period, 45 cases of alcoholic cirrhosis associated liver cancer (ALD group), and 71 cases of hepatitis B combined with alcoholic cirrhosis associated liver cancer (HBV+ALD group) were enrolled. At the same time, 73 patients with hepatitis B cirrhosis associated liver cancer (HBV group) during the same period were randomly selected with a ratio of about 1∶1.5, totaling 189 cases. And 183 (96.8%) of the patients were male and 6 (3.2%) were female. The age was (55.93±10.20) years. 109 deaths (57.7%) were recorded. The median survival times were 12 months for the entire cohort, 55 months for HBV group, 36 months for ALD group and 11 months for HBV+ALD group. And the 10-year death rate was 42.5% in HBV group, compared to 66.7% in ALD group and 67.6% in HBV+ALD group. In this study, 93 patients chose either the surgical resection or the radiofrequency ablation as their treatments. The recurrence rate was 69.9%, the median recurrence time was 8 months and the median overall survival time was 39 months. Univariate Cox regression identified that etiology of HBV and ALD, alpha-fetoprotein (AFP)>1 200 ng/mL, Child-Pugh class B and C, Barcelona Clinic Liver Cancer (BCLC) stages of C and D and curative therapies such as surgery and radiofrequency ablation were significantly correlated with overall survival (all P<0.05). Multivariate Cox regression revealed that patients with both HBV and ALD (HR=1.750，95%CI: 1.107-2.765，P=0.017), AFP>1 200 ng/mL (HR=1.649，95%CI: 1.060-2.564，P=0.027), and BCLC stages of C and D (HR=3.404，95%CI: 2.254-5.142，P<0.001) were independent risk factors of mortality in PLC patients with cirrhosis. Conclusions Among HBV, ALD and HBV+ALD groups, the HBV+ALD group had the shortest median survival time and the highest overall mortality rate, suggesting that alcohol consumption and HBV infection may accelerate the progression of PLC with cirrhosis and worsen its prognosis. HBV infection combined with alcoholic consumption, AFP>1 200 ng/mL, and BCLC stages of C and D were independent risk factors for mortality in PLC patients with cirrhosis.
Objective To analyze the clinical characteristics and prognostic factors for clinical cure in chronic hepatitis B virus (HBV) infected patients with pegylated interferon-α-2b (PEG-IFN-α-2b) based therapy. Methods This is a retrospective study. Chronic HBV infected patients receiving PEG-IFN-α-2b therapy were enrolled in Yuncheng Central Hospital affiliated to Shanxi Medical University between August 2020 and November 2023. Treatment-naïve patients received PEG-IFN-α-2b monotherapy, while nucleoside (acid) analogs (NAs)-experienced patients received PEG-IFN-α-2b add-on therapy. The study endpoint was hepatitis B surface antigen (HBsAg) negative in accompany with HBV DNA below the detection limit (course of treatment < 48 weeks) or treatment for 48 weeks. The general characteristics, virological variables, blood routine test, and liver function of patients were collected at baseline and study endpoint. Student’s t test or Mann-Whitney U test was used for comparison. The prognostic factors of clinical cure were examined using univariate and multivariate stepwise logistic regression models. Results A total of 61 chronic HBV infected patients were enrolled, including 39 males and 22 females. The age were (39.13±7.53) years. Twenty-one cases were treatment-naïve, while 40 patients were NAs-experienced. Nineteen cases were positive for HBV DNA at baseline. The baseline HBsAg levels were 211.30(50.93, 2 110.00) IU/mL. Thirty-four patients achieved clinical cure at the study endpoint with 25.50 weeks of median course of PEG-IFN-α-2b treatment. Twenty-seven patients did not achieve clinical cure at the study endpoint, and the course of treatment was 48 weeks. Clinical cure group had significant lower baseline HBsAg level when compared with non-clinical cure group [78.66(19.54, 204.60) IU/mL vs 2 078.00(442.20, 4 237.00) IU/mL, P<0.001]. At the study endpoint, the white blood cell, platelet, red blood cell, hemoglobin levels were lower than those in baseline (P<0.05), while alanine aminotransferase and asparatate aminotransferase levels were higher than those in baseline (P<0.05). There were no remarkable differences in blood routine tests or liver function between clinical cure and non-clinical cure group at either baseline or study endpoint (P>0.05). Baseline low HBsAg level was a predictor for clinical cure in patients receiving PEG-IFN-α-2b therapy (OR=0.998, 95%CI: 0.998-0.999). No severe adverse events were happened during therapy. Conclusion PEG-IFN-α-2b-based therapy was well-tolerant, and could achieve high rate of clinical cure in chronic HBV-infected patients who had low baseline HBsAg level.
Chronic hepatitis B virus (HBV) infection remains a major global public health concern. The persistent presence of intrahepatic covalently closed circular DNA (cccDNA) poses a major challenge to the complete cure of chronic HBV infection. Therefore, identifying reliable and effective serological surrogate markers for cccDNA holds great clinical significance in evaluating antiviral efficacy, predicting prognosis, and guiding the clinical management of chronic HBV-infected patients. In recent years, serum HBV RNA has emerged as a promising non-invasive alternative marker for cccDNA, offering the potential for monitoring disease progression and predicting prognosis in chronic HBV-infected patients. In this review, we summarize recent studies on HBV RNA, highlighting its ability to assess the immune and histological status of patients, and discussing its value in guiding the timing of antiviral therapy. Furthermore, we systematically summarize the clinical significance of HBV RNA in multiple domains: monitoring viral replication, evaluating antiviral treatment efficacy, predicting relapse after treatment cessation, and guiding new antiviral strategies. This review aims to provide clinicians with valuable insights for better utilizing this marker in the clinical management of chronic HBV infection.
Objective A predictive model should be established during the early stages of dengue progression to evaluate the likelihood of severe dengue and dengue with warning signs, thereby preventing delayed clinical management and reducing dengue-related mortality. Methods Clinical and laboratory examination data of 831 patients admitted to Ruili People's Hospital of Yunnan Province during 2019-2023 were retrospectively collected. The dataset was divided into a training set and a validation set in a 7∶3 ratio. Statistical description and univariate analysis were performed on the training set, with LASSO regression employed to screen variables, followed by logistic regression to develop a risk prediction model for severe dengue. Model performance was validated using ROC curves on both the training set and validation set. Results A total of 831 dengue patients were included in the study, with a mean age of (44.20±15.02) years. Among them, 52.59% were male and 5.42% were Myanmar nationality. In total, 122 cases (14.68%) exhibited severe dengue or dengue with warning signs, predominantly female (58.20%). LASSO regression was used in the training set to screen 11 variables related to the risk of severe dengue and dengue with warning signs: Age, dizziness, vomiting, prothrombin time, partial activated thromboplastin time, hematocrit, platelet, monocyte percentage, absolute value of monocytes, hemoglobin, C-reactive protein (λmin= 0.011 59); Logistic regression identified statistically significant variables for the risk model of severe dengue and dengue with warning signs as follows: age [OR=1.034 (95%CI: 1.016-1.053)], red blood cells deposited [OR=1.258 (95%CI: 1.143-1.519)], platelet [OR=0.991 (95%CI: 0.985-0.997)], hemoglobin (OR=0.919 (95%CI: 0.873-0.950)], C-reactive protein [OR=1.019 (95%CI:1.004-1.034)]. The model achieved an AUC of 0.894 (95%CI: 0.796-0.867) in the training set and 0.862 (95%CI: 0.709-0.827) in the validation set. At a cut-off threshold of 0.197, sensitivity and specificity were 0.850 and 0.743, respectively. Conclusion This study established a LASSO-logistic regression model, which can predict the risk of severe dengue and dengue with warning signs. The model enhances the capability of hospitals to prevent and manage severe dengue and provides valuable guidance for clinical decision-making.
Objective To analyze the epidemiological characteristics of varicella infectious diseases in Hainan Province from 2014 to 2023, and explore the epidemic patterns of varicella in Hainan Province. Methods Download data related to chickenpox in Hainan Province from the Infectious Disease Reporting Information Management System and the Public Health Emergency Management System of the Chinese Disease Control and Prevention Information System, descriptive epidemiological methods were used to analyze the incidence characteristics of varicella; ARIMA model was used to predict the incidence trend. Results From 2014 to 2023, a total of 65 832 cases of varicella and one death case were reported in Hainan, with an average annual incidence rate of 66.50/100 000, and the incidence rate fluctuated between 47.31/100 000 and 89.01/100 000. A incidence rate peak occurred every other year. The incidence of varicella showed a seasonal bimodal distribution, with peaks in January and April. The incidence of varicella was negatively correlated with temperature (r=-0.349，P<0.05). Children aged 0-14 years were the main incidence group, and the incidence rate reached its peak in the age group of 5-6 years. Students and young children were the main groups affected by the disease. The incidence rate of male (68.15/100 000) was higher than that of female (62.22/100 000)(χ2=16.80, P<0.05). The incidence rate of male (68.15/100 000) was higher than that of female (62.22/100 000). The 1 dose vaccination rate for 1-year-old children was on the rise, reaching a maximum of 43.2%; The two doses vaccination rate for children aged 4-6 years has been increasing year by year, with the highest reaching 9.9%. A total of 65 cases of varicella PHEE were reported, with M(P25, P75) of 24 (18, 32) cases per case, and a total incidence rate of 2.64%. Primary schools were the main places of occurrence. The ARIMA model has a good predictive fitting effect, and the results show that the predicted incidence in 2024 will increase compared to 2023. There may be peaks in incidence in January to June and October to December. Conclusions The incidence rate of varicella is high in Hainan Province; A lower vaccination rate for varicella in children cannot break the natural prevalence of varicella among healthy children.
Objective To investigate the resistance levels of adult Culex quinquefasciatus mosquitoes to three commonly used pyrethroid insecticides in Xingyi City, Guizhou Province, as well as to examine the changes in metabolic detoxification enzyme activities and mutations in the knockdown resistance (kdr) gene thereby providing a scientific foundation for the prevention and control of Culex quinquefasciatus in the region. Methods In 2024, the larvae of Culex quinquefasciatus were collected from different locations in Xingyi and reared to adult mosquitoes in the laboratory, and their resistance to pyrethroid insecticides was determined using the adult mosquito contact tube method. Surviving samples (resistant population) from the resistance tests were examined for the activities of mixed function oxidase (MFO), glutathione-S-transferase (GST), and non-specific esterase (NSE) using a microplate reader. The genomic DNA of individual adult Culex quinquefasciatus mosquitoes was extracted, and the voltage-gated sodium channel (VGSC) gene fragment was amplified via PCR and sequenced to analyze the mutations in the kdr gene. Results The 24-h mortality rates of adult Culex quinquefasciatus exposed to 0.25% permethrin, 0.025% deltamethrin, and 0.025% beta-cypermethrin were 1.00%, 0%, and 0.88%, respectively, indicating all populations of Culex quinquefasciatus exhibited resistance to the three pyrethroid insecticides. The activity of MFO in permethrin-resistant population followed a normal distribution, while deltamethrin-resistant and cypermethrin populations exhibited skewed distributions. The resistance ratios (RRs) for the three populations were 1.17, 1.03, and 1.07 times, respectively, with no statistically significant differences in MFO activity between field populations and susceptible strains (all P>0.05). However, the GST activities of permethrin-resistant and beta-cypermethrin-resistant field populations were 1.06 and 1.45 times higher than those of sensitive strains (P<0.05). Sequencing of the VGSC gene fragments of adult Culex quinquefasciatus resistant to pyrethroid insecticides revealed that mutations occurred solely at 1014 locus, presenting two alleles: wild type TTA(L) (1.26%) and mutant TTT(F) (98.74%). Two genotypes were identified: wild/mutant heterozygotes L/F (2.52%) and mutant homozygotes F/F (97.48%). No statistically significant differences were observed in the frequency of resistance gene mutations among populations resistant to the three pyrethroid insecticides (P>0.05). Conclusion The adult mosquitoes of Culex quinquefasciatus in Xingyi have developed resistance to three commonly used pyrethroid insecticides, with changes observed in metabolic detoxification enzyme activities and mutations in the kdr gene. The resistance of Culex quinquefasciatus in Xingyi is the result of multiple mechanisms. Understanding the resistance level and mechanism can provide a reliable basis for mosquito control and resistance management. It is recommended to enhance the monitoring of mosquitoes' resistance to insecticides, prioritize environmental management focused on eradicating breeding sites, reduce the use of pyrethroid insecticides, and rotate with other classes of insecticides to delay the onset of resistance.
Objective To construct a recombinant lentiviral expression vector for human lymphocyte activation gene 3 (LAG3) and generation of monoclonal cell lines that preferentially express LAG3 by transfection of the vector into mouse fibroblast cells 3T3. Methods After extracting total RNA extracted from human peripheral blood mononuclear cells, the RNA is reversely transcribed into cDNA. The LAG3 extracellular and transmembrane region sequences are amplified by PCR using high-fidelity DNA polymerase. The PCR products are double-digested with the restriction endonucleases EcoRⅠ and NotⅠ, then ligated with the lentiviral vector pTSB-copGFP to construct the recombinant expression vector pTSB-LAG3-copGFP, which is subsequently transformed into Escherichia coli DH5α. Positive clonal bacteria are selected by PCR, and the plasmids are extracted and sequenced for verification. The recombinant vector pTSB-LAG3-copGFP, along with packaging plasmids psPAX2 and pMD2.0G, are co-transfected into human embryonic kidney 293T cells to assemble and release virus particles, the virus infected 3T3 cells were collected. During the puromycin selection of infected 3T3 cells, the limited dilution method is used to obtain 3T3 monoclonal cells that stably express LAG3. Real-time fluorescent quantitative PCR, immunofluorescence and flow cytometry were utilized to verify the transcription of LAG3 mRNA and the expression of LAG3 protein respectively. Results Sequencing of the recombinant pTSB-LAG3-copGFP lentiviral vector plasmid reveals that the amplified LAG3 sequence contains a synonymous mutation in the His codon at nucleotide position 1 697 bp within the LAG3 transmembrane region, which aligns with the standard LAG3 sequence (accession number NM_002286.6) in GenBank. The 3T3 cells infected by pTSB-LAG3-copGFP packaging virus screened with puromycin. A total of 20 LAG3+copGFP+-3T3 monoclonal cell lines were obtained, all of which exhibited transcription of LAG3 mRNA. The monoclonal cell line MC-6 exhibits the highest transcriptional level of LAG3. Effective expression and distribution of LAG3 protein on the cell membrane and cytoplasmic organelle membranes in MC-6 indicated by immunofluorescence and flow cytometry. Conclusion The pTSB-LAG3-copGFP lentiviral vector was successfully constructed. LAG3+copGFP+-3T3 monoclonal cell lines overexpressing lymphocyte activating 3 were efficiently established, laying the foundation for subsequent studies on the relationship between LAG3 and the development of chronic infectious diseases such as hepatitis B, as well as the interventional treatment of LAG3.
Objective To explore the whole genome structure of Brucella in Ordos city, conduct bioinformatics analysis, and explore a variety of Brucella species type identification methods, so as to provide a theoretical basis for the study of Brucella in Ordos City. Methods The whole genome was sequenced using illumina Miseq gene sequencer. The Microobench microbial gene analysis workstation was used to analyze the genome components, genome annotation, 16Sr DNA, SNP sites, insertions and deletions, MLST sites, and phylogenetic tree of 15 strains of Brucella. Results The average sequencing depth of 15 Brucella strains was 170.04×, the average genome size was 3 290 725 bp, the genome coverage was 99.93%, and the average GC content of assembly files was 54.69%. The results of 16Sr DNA analysis showed that the homology of 15 Brucella strains and Brucella Maltese was 100%. The analysis results of SNP sites and insert-deletion variation showed that the average number of SNPs of the 15 strains of Brucella was 2 412, the average number of polynucleotide mutations was 39, the average number of insertional variants was 131, the average number of missing variants was 153, the average number of complex variants was 43, and the average number of total variants was 2 778. MLST locus analysis showed that all 15 strains of Brucella were ST-8. Conclusion Based on the whole genome sequencing of 15 strains of Brucella in Ordos City, this study carried out a variety of type identification methods. Different methods have different focuses, and multiple methods are needed to confirm each other, so as to obtain more accurate experimental results and provide more research basis for the study of Brucella, to provide more data and theoretical support for the control and prevention of brucellosis in Ordos City.
Objective To understand and grasp the status quo of resistance of Culex pipiens pallens to four commonly used insecticides in Hefei City, and to provide a scientific basis for the chemical control of mosquito larvae. Methods From June to July 2023, Cx. pipiens pallens larvae were collected from 9 counties (cities and districts) in Hefei City. The LC50 of late third-instar to early fourth-instar larvae of Cx. pipiens pallens to commonly used insecticides was determined by larval immersion method (sensitive baseline method). Results Cx.pipiens pallens larvae in Hefei City exhibited different degrees of resistance to four insecticides: permethrin, beta-cypermethrin, temephos, and propoxur. The relative resistance coefficients to permethrin and beta-cypermethrin were 26.96 and 21.17, respectively, indicating the moderate resistance level. The relative resistance coefficients to propoxur were 6.70, indicating a low resistance level. The relative resistance coefficient to temephos was 2.43, indicating a sensitivity level. Culex pipiens pallens against pyrethroids such as 0.25% permethrin, 0.025% deltamethrin and 0.025% cypermethrin in 1 h knockout rate and 24 h mortality rates were 3.25% (4/123) and 46.34% (57/123), 3.60% (5/139) and 35.97% (50/139), 3.85% (6/156) and 40.38% (63/156), respectively. For 5% malathion and 0.1% propoxur, the 1 h knockdown rate and 24 h mortality rate were 97.69% (127/130) and 99.23% (129/130), 94.48% (137/145) and 100.00% (145/145), respectively. It showed resistance to 0.25% permethrin, 0.025% deltamethrin and 0.025% cypermethrin, and sensitivity to 5% malathion and 0.1% propoxur. Conclusions Culex pipiens pallens in Hefei City have developed varying degrees of resistance to parathyroid and carbamate insecticides. In the control of mosquito vectors, it is essential to strengthen the scientific and rational use of chemical control in combination with environmental and physical control measures to form an integrated control strategy. This approach will improve the control efficiency while delaying the occurrence and development of insecticide resistance.
Objective To detect clustered regularly interspaced short palindromic repeats (CRISPR) in Listeria monocytogenes, and analyze the structure and homology of CRISPR loci. Methods Totally 34 strains of Listeria monocytogenes isolated in our laboratory were identified, PCR amplified and sequenced. The repeat sequence structure and spacer sequence homology in CRISPR loci were analyzed by bioinformatics software. Results A total of 7 CRISPR loci were detected in 34 strains. The mutation rate of the first 2 and last 2 bases of the Repeat sequence of CRISPR loci was higher, while the mutation rate of the middle part was lower. Seven CRISPR sites form eight CRISPR structural types, among which the Repeat sequences of CRISPR1 and CRISPR2 are relatively conserved, while the Repeat sequences of CRISPR1 and CRISPR5 can form dumbbell shaped secondary structures. The number of Spacer sequences contained in each CRISPR site ranges from 2 to 15, with an average of 2.43. The 136 Spacer sequences detected were not only homologous to Listeria plasmids and bacteriophages, but also homologous to uncultured virus sequences, staphylococcal bacteriophages, and Listeria innocua. The same CRISPR genotype did not show large-scale clustering, but some strains in the same year were in the same evolutionary cluster with close genetic relationships. Conclusion The CRISPR structure of Listeria monocytogenes in this study exhibits high specificity, and its homology with bacteriophages provides a theoretical basis for the application of bacteriophages in the control and prevention of Listeria monocytogenes.
Objective To conduct an in-depth study of the molecular biological characteristics and evolutionary trends of H9N2 avian influenza virus (AIV) in live poultry markets in Yunnan Province in 2023, and to provide scientific evidence for the development of control strategies for H9N2 avian influenza in the region. Methods Environmental samples were collected from live poultry markets in Yunnan Province in 2023 for H9N2 subtype nucleic acid detection. Positive samples were subjected to virus isolation using chicken embryos, and the genome of the 8 isolated strains was amplified, sequenced, and analyzed for genetic characteristics. Results The eight avian influenza virus (AIV) isolates had the hemagglutinin (HA) cleavage site sequence PSRSSRGLF, which is a non-continuous basic amino acid sequence, consistent with the genetic characteristics of typical low-pathogenicity avian influenza viruses. Mutations Q234L and H191N were observed in the left arm of the HA protein, which enhanced the affinity for α-2,6 sialic acid receptors, suggesting that these viruses may have the potential to infect humans. The neuraminidase (NA) protein exhibited a deletion of three amino acids (TEI) at positions 62-64 in the stalk region, displaying characteristics of high pathogenicity at the molecular level. The increase or absence in potential glycosylation sites were observed in both HA and NA genes. The non-structural protein 1 (NS1) showed no D92E mutation, and had a C-terminal truncation of 13 amino acids, indicating that this virus is of low pathogenicity and poses a lower risk of human transmission. Mutations T37A, R95K, S224N, and K242N in the M1 protein of some isolates increased the risk of infection, while one isolate carried the V27A or S31N mutation in the M2 protein, conferring resistance to M2 ion channel inhibitors. Mutations M317I and S678N were identified in the PB1 protein, which may enhance pathogenicity in mice and increase the potential for mammalian infection. The PB2 protein carried the I292V mutation, which exhibited a stronger infectivity to mammals. Phylogenetic analysis revealed that the HA, NA, and PB2 gene segments belonged to the Y280 lineage, NP and PB1 gene segments were classified under the F98 lineage, the M gene segment of the NH013 isolate belonged to the F98 lineage, while M genes, as well as the NS and PA genes of other isolates belonged to the G1 lineage. Conclusions These eight AIV isolates exhibited characteristics of low pathogenicity, but simultaneously carry the potential risk of infecting humans. Despite the HA cleavage site and NS1 protein mutations indicating low pathogenicity, the Q234L and H191N mutations in the HA protein enhanced its affinity for human receptors, suggesting the potential for human infection. The TEI deletion in the NA protein, mutations in the M1 protein, and resistance mutations in the M2 protein further increase the risk of human infection. Mutations in the PB1 and PB2 proteins increase the potential for these eight AIV strains to infect humans or mammals.
Objective To analyse the crude detection rate and trends of newly detected HIV/AIDS cases in medical institutions in China from 2017 to 2023, and to provide a reference for optimizing HIV testing strategies in medical institutions. Methods Data on HIV testing and newly reported HIV/AIDS cases were analysed using data from the Comprehensive AIDS Prevention and Control Information System of the China Information System for Disease Control and Prevention for the period from 2017 to 2023. HIV testing in medical institutions includes patients tested preoperatively, those tested before transfusion, those tested in sexually transmitted disease (STD) clinics, prenatal care clinics, and other types of patients. Descriptive statistical analysis and χ2 test were performed using SAS 9.4 software. Joinpoint regression was performed using Joinpoint 4.9.0 software to analyse trends of the crude detection rates over time. Results From 2017 to 2023, the person-times of HIV tests in medical institutions increased from 143 million to 255 million, with an increase of 78.07%. The number of newly detected HIV/AIDS cases increased from 74 000 to 88 000 and then declined to 69 000. The crude detection rate of new HIV/AIDS cases declined from 5.18/10 000 to 2.71/10 000, showed a declining trend, the mean annual percentage change was -9.99%(P<0.001). The crude detection rate of new HIV/AIDS cases in STD clinics was the highest among all types of clinic visits (12.79/10 000-24.47/10 000), and the crude detection rate of new cases among all types of clinic visits showed a decreasing trend(P<0.05). Among different medical institutions, general hospitals were the most important source of the number of tests and the number of newly detected HIV/AIDS cases, accounting for more than 62.93% and 62.68%, respectively. Specialised medical institutions had the highest crude detection rate of new cases, which was maintained at more than 5.13/10 000. The crude detection rate of new cases for all four types of medical institutions, except for primary medical institutions, showed a decreasing trend (P<0.05). Conclusions The detection rate of new cases in medical institutions showed a decreasing trend in 2017-2023, and the efficiency of STD clinics testing and detection was higher among all types of attendees. General hospitals are the main source of new cases detection, and testing in specialised medical institutions is more efficient. Testing should be strengthened in key groups of patients and in key medical institutions.
Objective To investigate the distribution and carriage of Bartonella in rodents and plateau pikas (Ochotona curzoniae) in Xining City, Hai Prefecture, and Menyuan County in Qinghai Province, providing basic data for the prevention and control of Bartonella infections and scientific evidence for the control natural zoonotic diseases in the local area. Methods In 2023, rodents were captured using rodent traps across Xining City, Hai Prefecture, and Menyuan County in Qinghai Province. Bartonella DNA was extracted and amplified from collected liver, spleen, and kidney tissue samples of plateau pikas. Statistical analysis was performed using SPSS 28.0 software, with the chi-square test and Fisher's exact probability method employed to analyze Bartonella positivity rates across different rodent species and ecological environments. Sequence analysis was also conducted on the Bartonella-positive strains from Menyuan County. Results A total of 489 rodents and plateau pikas were captured across the three regions: 206 in Xining City, 200 in Haixi Prefecture, and 83 in Menyuan County. The Bartonella positivity rates were 40.29% (83/206), 2.00% (4/200), and 12.05% (10/83) respectively. A statistically significant difference in Bartonella positivity was observed between Menyuan County and Haixi Prefecture (χ2=10.55, P<0.01). In Menyuan County, 74 Mus musculus linnaeus were captured, 9 Ochotona were captured, 10 Mus musculus linnaeus were Bartonella positive, Ochotona was not detected, the difference was statistically significant by Fisher's exact probability test (P<0.05). There were 14 and 69 rodents captured on grassland and stone land, respectively. 10 rodents captured on stone land were positive for Bartonella, but no rodents were detected on grassland, and the difference was statistically significant by Fisher's exact probability test (P<0.05). Genomic sequence analysis showed that the 10 Bartonella strains from Menyuan County clustered into three genotypes:Bartonella sp.AR 15-3, Bartonella grahamii as4aup, and Bartonella grahamii subsp. shimonis. Conclusion Natural Bartonella infections exist in rodents and plateau pikas across Xining City, Haixi Prefecture, and Menyuan County in Qinghai Province. Furthermore, gene types related to human pathogenicity pose a risk of human infection. Therefore, the monitoring of rodent-borne pathogens in these areas should be strengthened.
Objective To further enrich the resources and Ceratopogonidae species in Beijing, an investigation and collection of biting midges in Lianshihu Wetland Park, Beijing, were carried out, leading to the description of two new species of the genus Dasyhelea in Beijing. Methods In September 2019, biting midges were collected in Lianhu Wetland Park, Beijing, using the net sweeping methods. The specimens were preserved in 75% alcohol for identification according to their morphological characteristics such as antennae and terminalia. Results A total of five species of biting midges belonging to the genus Dasyhelea were collected, including two new species: Dasyhelea extraneus Liu et Yu, sp. nov. and D. bibacula Liu et Yu, sp. nov. The main distinguishing characteristics of the two new species are as follows: In Dasyhelea extraneus, there is a tuberculate process in the middle of the posterior margin of the ninth sternite of the terminalia, the parameral lobes are fused into a bony strip with a short median process, and the aedeagal mesophallus has an obvious median process. In Dasyhelea bibacula, the posterior margin of the tergite of the terminalia is bluntly rounded, the aedeagal mesophallus is penholder-like, the ends of the two lateral branches are curved and fist-like, and the end of the median process of the parameral lobe is bluntly rounded. Conclusions This study describes two new species of the genus Dasyhelea in Beijing, enriching the midge resources in Beijing. The type specimens of the two new species are currently preserved at Nanchang Center for Disease Control and Prevention.
Objective To analyze the epidemiological characteristics and infection pathways and virulence factors of one cholera outbreak in Fengxian District, Shanghai, China, in 2024, and to provide scientific basis for epidemic control and prevention. Methods Epidemiologic data of cases of one cholera outbreak in Fengxian District in 2024 were collected using on-site epidemiologic survey methods; RT-PCR nucleic acid testing and bacterial culture were applied to carry out pathogenicity testing of cases, close contacts, environment, and food samples; and the genome sequences of the strains were obtained using second-generation gene sequencing. Results The case was a 62-year-old woman, who presented to the doctor with diarrhea for 4 consecutive days, 4-5 times a day, with watery stools, which was not effectively relieved by self-administered medication. There was no history of traveling away from Shanghai for 5 days before the onset of the disease, and she was engaged in the preparation and delivery of food for rural banquets during the period. Vibrio cholerae O139 was detected in the anal swab sample of the case and the septic tank of the workplace on the 4th day after the onset of the disease; samples of turtle and links in the store selling turtle were cultured for Vibrio cholerae O139. The isolate carried several virulence-related genes such as ctxA, ctxB, HlyA, zot, rtxA, hapA, nanH, tdh, and T3SS. Comparison of the isolate with the O139 strain of cholera cluster within our country through the National Pathogenic Bacteria Recognition Network (NPBN) in recent years suggests that the closest environmental or aquatic animal isolate to the sequence of this strain is the turtle isolate uploaded at a place in Guangdong. Conclusion This outbreak was a disseminated outbreak caused by the case's contact with turtle contaminated with Vibrio cholerae O139, and early detection of enteric infectious diseases such as cholera can be achieved by relying on the outpatient enteric cholera surveillance network.
Tropical diseases pose a significant challenge to global public health. Their epidemiological footprints have expanded due to global climate warming, population migration, and lifestyle transformations, affecting even non-tropical areas. "One Health" emphasizes interdisciplinary, intersectoral, and interregional cooperation and communication, which is of critical importance in the prevention and control of tropical diseases. Tropical disease education constitutes an essential component of preventive medicine education; however, it faces outdated content and methods that fail to keep up with the rapid changes in tropical disease scenarios and emerging prevention needs. In 2019, the Ministry of Education launched the "Double Ten Thousand Plan" to establish national and provincial first-class undergraduate disciplines, aiming to improve the quality of undergraduate education comprehensively. Building a first-class preventive medicine program bears the responsibility of cultivating high-level public health professionals. This paper aims to develop a "Tropical Medicine" course tailored to national conditions while highlighting local characteristics. The teaching process is reformed in four aspects: actively integrating the One Health concept, setting up professional teaching institutions, constructing virtual simulation laboratories, and strengthening academic exchanges and cooperation, specific suggestions are given in this regard.
To analyze the investigation and disposal of an H5N6 human infection with animal derived influenza epidemic in Quanzhou City in 2024, and provide reference for the control and prevention of animal derived influenza epidemics in future.According to Technical Guidelines for Control and Prevention of Human infected with Zoonotic Influenza Viruses (Trial), we investigated the diagnosis and treatment of the case, suspected exposure, infection sources and routes, by the method of field epidemiological investigation. The case, close contacts, and relevant environmental samples were collected for nucleic acid testing and gene sequencing. A nasopharyngeal carcinoma patient who rapidly progressed to severe pneumonia and died of multiple organ failure after being diagnosed with H5N6. The H5N6 virus nucleic acid test results of 5 close contacts and 37 suspected exposed individuals were all negative, and no second-generation cases occurred. Through on-site investigation and genetic sequencing analysis, it is highly likely that the source of infection is the outdoor environment of free range poultry contaminated with H5N6 virus in the vicinity of the patient's home.Patients with nasopharyngeal carcinoma or other immune restricted diseases should take personal protective measures, stay away from poultry and livestock breeding and trading places, and avoid close contact with poultry or other animals as much as possible. We should strengthen the training of management personnel of third-party testing institutions, enhance the work of thematic risk assessment and the application of risk assessment results,and expand the scope of monitoring human infected with zoonotic influenza viruses in breeding farms, live poultry markets, and migratory bird habitats.
