Distributive characteristics of HBV DNA CpG islands in HBsAg positive mothers and its relationship with intrauterine transmission / 中华流行病学杂志

Objective: To investigate the type, length, and CG loci of HBV DNA CpG islands in HBsAg positive maternal C genotype and its relationship with intrauterine HBV transmission, so as to provide a new perspective for the study of intrauterine transmission of HBV. Methods: From June 2011 to July 2013, HBsAg-positive mothers and their newborns who delivered in the obstetrics and gynecology department of the Third People's Hospital of Taiyuan were collected. Epidemiological data were collected through face-to-face questionnaires and electronic medical records. Serum HBV markers and serum HBV DNA were detected by electrochemiluminescence and quantitative fluorescence PCR, respectively. Intrauterine transmission of HBV was determined by positive HBsAg and/or HBV DNA in femoral venous blood before injection of HBV vaccine/Hepatitis B immunoglobulin within 24 h of birth. A total of 22 mothers and their newborns with HBV DNA load ≥106 IU/ml in intrauterine transmission were selected as the intrauterine transmission group, and 22 mothers with HBV DNA load ≥106 IU/ml without intrauterine transmission were chosen as the control group by random seed method. The distribution prediction of CpG islands of HBV DNA in 39 mothers with genotype C by HBV DNA sequencing was analyzed. Results: Among 39 mothers with HBV C genotype, 19 were in the intrauterine transmission group, and 20 were in the control group. The HBV DNA of 39 patients with genotype C traditional CpG island Ⅱ and Ⅲ, while the control group had traditional CpG island Ⅰ and novel CpG island Ⅳ and Ⅴ. The length of CpG island Ⅱ and Ⅲ and the number of CG loci of CpG island Ⅱ in the intrauterine transmission group differed from those in the control group (P<0.05). The CpG island Ⅱ length ≥518 bp and the number of CG loci ≥40 in the intrauterine transmission group (11/19) were significantly higher than those in the control group (2/20) (P<0.05). The length of CpG island Ⅱ and the number of CG loci in the X gene promoter region (Xp region) were higher than those in the control group (P<0.05). In the HBV intrauterine transmission group, most of maternal (12/19) HBV DNA CpG island Ⅱ completely covered the Xp region, which was significantly higher than that in the control group (5/20), and the number of HBV DNA Xp region CG loci was higher than that in the control group (P<0.05). Conclusions: The distribution of maternal C genotype HBV DNA CpG islands is related to intrauterine transmission. The length of CpG island Ⅱ and the number of CG sites may increase the risk of intrauterine transmission of HBV.

Non/hypo-response to hepatitis B vaccination and influencing factors in HIV-infected patients in the context of different immunization schedules / 中华流行病学杂志

Objective: To study the non/hypo-response to hepatitis B vaccination in HIV-infected patients, identify the influencing factors and provide evidence for the development of hepatitis B prevention and control strategies and measures for special population. Methods: On the basis of the randomized controlled trial of 20 µg hepatitis B vaccine immunization at 0-1-6 month, 0-1-2-6 month and 60 µg hepatitis B vaccine immunization at 0-1-2-6 month, the HIV-infected patients who completed one-month follow-up after the full course vaccination were selected as study subjects. Quantification of antibody to hepatitis B surface antigen (anti-HBs) in serum samples was performed by using chemiluminescent microparticle immunoassay (CMIA) and demographic characteristics, disease history, HIV infection and treatment status of the study subjects were collected. Statistical analysis was conducted by χ2 test, t test, unconditional logistic regression and interaction analyses. Results: The non/hypo-response rates to hepatitis B vaccination were 34.65% (35/101), 24.49% (24/98) and 10.99% (10/91) in 20 µg group at 0-1-6 month or 0-1-2-6 month and 60 µg group at 0-1-2-6 month (P<0.001), respectively. Logistic regression analysis showed that after controlling for confounding factors, the risk for non/hypo-response was 0.22 times higher in HIV-infected patients receiving 60 µg hepatitis B vaccine at 0-1-2-6 month than in patients receiving 20 µg hepatitis B vaccine at 0-1-6 month (95%CI: 0.10-0.50), the risk for non/hypo-response was higher in men than in women (OR=3.65, 95%CI: 1.88-7.07), and the risk for non/hypo-response was 2.64 times higher in those without hepatitis B vaccination history than in those with hepatitis B vaccination history (95%CI: 1.10-6.32). Moreover, there were multiplicative interactions between immunization schedule and gender (OR=2.49, 95%CI: 1.24-5.00). Conclusion: The non/hypo-response rate to hepatitis B vaccination was significantly lower in HIV-infected patients receiving 60 µg hepatitis B vaccine at 0-1-2-6 month than in those receiving 20 µg hepatitis B vaccine at 0-1-6 month and 0-1-2-6 month. Gender, vaccination schedule and history of hepatitis B vaccination were the influencing factors of the non/hypo-response to hepatitis B vaccination. There was a multiplicative interaction between vaccination schedule and gender, and men receiving 20 µg hepatitis B vaccines had a higher risk for non/hypo-response to hepatitis B vaccination.

Effect of activation of Toll-like receptor signaling pathway of peripheral blood mononuclear cell in recombinant hepatitis B surface antigen immune response / 中华流行病学杂志

Objective: To explore the effect and mechanism of activation of peripheral blood mononuclear cell (PBMC) Toll-like receptor (TLR3) signaling pathway in recombinant HBsAg (rHBsAg) immune response. Methods: White blood cells were collected from peripheral blood of 13 healthy donors in the preparation of blood products. PBMC was isolated and treated with Poly I:C (Poly I:C group) and PBS (control group) respectively. 48 h later, some cells were collected and the expressions of TLR3 signaling pathway proteins were detected by flow cytometry. After activating (Poly I:C group)/inactivating (control group) TLR3 signaling pathway, rHBsAg was given to both groups for 72 h, and the proportions of DC, T, B cells and their subsets in PBMC were detected by flow cytometry. Paired t-test, paired samples wilcoxon signed-rank test and canonical correlation analyses were used for statistical analysis. Results: The percentage of TLR3 protein-positive cells (19.21%) and protein expression (8 983.95), NF-κB protein expression (26 193.13), the percentage of pNF-κB protein-positive cells (13.73%) and its proportion in NF-κB (16.03%), and the percentage of pIRF3 protein-positive cells (12.64%) and its proportion in IRF3 (21.80%) in Poly I:C group were higher than those in control group (11.54%, 8 086.00, 22 340.66, 8.72%, 9.71%, 9.57%, 19.12%) (P<0.05), and the percentage of TRIF protein-positive cells (89.75%) and protein expression (304 219.54) were higher in Poly I:C group than in the control group (89.64%, 288 149.72) (P>0.05). After PBMC stimulation by rHBsAg, the proportions of mDC (2.90%), pDC (1.80%), B cell (5.31%) and plasma cell (67.71%) in Poly I:C group were significantly higher than those in the control group (1.83%, 0.81%, 4.23%, 58.82%) (P<0.05). Results of canonical correlation analysis showed that the expression of TLR3 protein was positively correlated with the proportions of plasma cells, the expression of pIRF3 protein was positively correlated with the proportions of plasma cells and mDC, and the percentage of pNF-κB protein-positive cells and the percentage of pIRF3 protein-positive cells were positively correlated with the proportion of CD4+T cells. Conclusions: Poly I:C can activate TLR3/TRIF/NF-κB and TLR3/TRIF/IRF3 signaling pathway, promote the function of downstream signaling molecules, and then promote the maturation of DC, induce the immune responses of CD4+T cell, and promote the maturation and activation of B cells and the immune response of rHBsAg.

Immunogenicity and influencing factors of hepatitis B vaccination based on different vaccination schedules among chronic kidney disease patients / 中华流行病学杂志

Objective: To explore the immunogenicity and influencing factors of hepatitis B vaccination based on different vaccination schedules among chronic kidney disease (CKD) patients. Methods: CKD patients who participated in randomized controlled trials in four hospitals in Shanxi province and completed three doses of 20 µg vaccination (at months 0, 1 and 6) and four doses of 20 µg or 60 µg vaccination (at months 0, 1, 2, and 6) were surveyed from May 2019 to July 2020.According to the ratio of 1∶1∶1, 273 CKD patients were divided into 3 groups randomly. Quantification of the anti-hepatitis B surface antigen-antibody (anti-HBs) in serum samples was performed using chemiluminescent microparticle immunoassay at months 1 and 6 after the entire course of the vaccinations. The positive rate, high-level positive rate, geometric mean concentration (GMC) of anti-HBs, and the influencing factors were analyzed by χ2 tests, analysis of variance, unconditional logistic regression analysis. Results: A total of 273 CKD patitents were participants.The positive rates in the CKD patients with four doses of 20 µg vaccination (92.96%,66/71) or 60 µg vaccination (93.15%, 68/73) were higher than that in the CKD patients with three doses of 20 µg vaccination (81.69%, 58/71) at month one after the full course of the vaccinations (P<0.05). The GMCs of anti-HBs showed similar results (2 091.11 mIU/ml and 2 441.50 mIU/ml vs. 1 675.21 mIU/ml) (P<0.05). The positive rate was higher in the CKD patients with four doses of 60 µg vaccination (94.83%,55/58) than in those with three doses of 20 µg vaccination (78.79%,52/66) (P<0.05) at month six after the full course of the vaccinations. And the GMC of anti-HBs in the patients with four doses of 60 µg vaccination (824.28 mIU/ml) was significantly higher than those in the patients with 3 or 4 doses of 20 µg vaccination (639.74 mIU/ml and 755.53 mIU/ml) (P<0.05). After controlling the confounding factors, the positive rate in the CKD patients with four doses of 60 µg vaccination were 3.19 (95%CI: 1.02-9.96) and 5.32 (95%CI: 1.27-22.19) times higher than those in the patients with three doses of 20 µg vaccination at months 1 and 6 after the full course of the vaccinations, respectively. The positive rate in CKD patients without immune suppression or hormone therapy was 3.33 (95%CI: 1.26-8.80) and 4.78 (95%CI: 1.47-15.57) times higher than those in the patients with such therapy, respectively. Conclusions: Four doses of 20 µg or 60 µg hepatitis B vaccination could improve the immunogenicity in patients with CKD. And four doses of 60 µg vaccination might play a positive role in maintaining anti-HBs in this population. The immunogenicity in the CKD patients with immune suppression or hormone therapy was poor.

Analysis on influencing factors of HBV intrauterine transmission based on integration of decision tree model and logistic regression model / 中华流行病学杂志

Objective: To investigate the influencing factors of HBV intrauterine transmission and their interaction effects by integrating logistic regression model and Chi-squared automatic interaction detector (CHAID) decision tree model. Methods: A total of 689 pairs of HBsAg-positive mothers and their neonates in the obstetrics department of the Third People's Hospital of Taiyuan from 2007 to 2013 were enrolled, and the basic information of mothers and their neonates were obtained by questionnaire survey and medical record review, such as the general demographic characteristics, gestational week and delivery mode. HBV DNA and HBV serological markers of the mothers and newborns were detected by fluorescence quantitative PCR and electrochemiluminescence immunoassay respectively. The CHAID decision tree model and unconditional logistic regression analysis were used to explore the factors influencing HBV intrauterine transmission in neonates of HBsAg-positive mothers. Results: Among the 689 neonates, the incidence of HBV intrauterine transmission was 11.47% (79/689). After adjusted for confounding factors, the first and second logistic multivariate analysis showed that cesarean delivery was a protective factor for HBV intrauterine transmission (OR=0.25, 95%CI: 0.14-0.43; OR=0.27, 95%CI: 0.15-0.46); both models indicated that maternal HBeAg positivity and HBV DNA load ≥2×105 IU/ml before delivery were risk factors of HBV intrauterine transmission (OR=3.89, 95%CI: 2.32-6.51; OR=3.48, 95%CI: 2.12-5.71), respectively. The CHAID decision tree model screened three significant factors influencing HBV intrauterine transmission, the most significant one was maternal HBeAg status, followed by delivery mode and maternal HBV DNA load. There were interactions between maternal HBeAg status and delivery modes, as well as delivery mode and maternal HBV DNA load before delivery. The rate of HBV intrauterine transmission in newborns of HBeAg-positive mothers by vaginal delivery increased from 19.08% to 29.37%; among HBeAg-positive mothers with HBV DNA ≥2×105 IU/ml, the rate of HBV intrauterine transmission increased to 33.33% in the newborns by vaginal delivery. Conclusions: Maternal HBeAg positivity,maternal HBV DNA ≥2×105 IU/ml and vaginal delivery could be risk factors for HBV intrauterine transmission in newborns. Interaction effects were found between maternal HBeAg positivity and vaginal delivery, as well as vaginal delivery and high maternal HBV DNA load. Logistic regression model and the CHAID decision tree model can be used in conjunction to identify the high-risk populations and develop preventive strategies accurately.

Establishment and verification of preeclampsia risk assessment model / 中华疾病控制杂志

Objective To analyze the risk factors affecting pre-eclampsia, to establish a pre-eclampsia risk assessment model, and to assess the risk of pre-eclampsia early. Methods A face-to-face questionnaire survey was conducted for all women who gave birth in the Department of Obstetrics, the First Hospital of Shanxi Medical University from March 2012 to September 2016. A total of 10 319 qualified questionnaires were collected to exclude 9 623 cases of other hypertensive diseases related to pregnancy. A total of 70% of the subjects were randomly selected as training samples to analyze the influencing factors of pre-eclampsia, and a Logistic regression model was established. The remaining 30% of the objects are used as test samples to verify the effect of the model. Results Logistic regression model was established with training samples. Logit P=-2.517-0.696×Pre-pregnancy lean +0.200 ×Pre-pregnancy overweight +0.944×Pre-pregnancy obesity -1.995×Residential in city -0.409×Folic acid supplemented before pregnancy +1.323×Twin and multiple pregnancy +1.708× History of previous pregnancy hypertension. Homer-Lemeshow test P=0.377. Model AUC=0.767 (95%CI:0.747-0.786, P<0.001). Using the test sample to verify the model, the model sensitivity was 81.68%, the specificity was 75.05%, the positive likelihood ratio was 3.27, and the negative likelihood ratio was 0.24. The test sample model AUC = 0.771 (95%CI=0.763-0.790,P<0.001). Conclusion This study establishes a simple and effective pre-eclampsia risk assessment model with controllable factors. The model has good fit and sensitivity and specificity.

Effect of PBMC HBV cccDNA in HBsAg-positive mothers on neonatal Th1/Th2 cytokines / 中华疾病控制杂志

Objective To explore the effect of PBMC HBV cccDNA in HBsAg-positive mothers on neonatal Th1, Th2 cytokines and the ratio of Th1/Th2. Methods HBsAg-positive mothers and their neonates delivered in the Third People’s Hospital of Taiyuan between June 2011 and July 2013 were recruited. Questionnaires on general information were collected by an in-person interview. Electrochemiluminescence immunoassay (ECLIA) were utilized to detect HBV serological markers.HBV cccDNA in PBMC was detected with real-time PCR-TaqMan Probe method, Th1 cytokines (interleukin 2, interferon-γ and tumor necrosis factor-α) and Th2 cytokines (interleukin 4, interleukin 6 and interleukin 10) were detected with Procarta Plex Multiplex Immunoassays. Results Univariate analysis showed that the levels of IL-2, IL-6 and IL-10 in the positive group were significantly higher than those in the negative group, while the ratio of Th1/Th2 was lower than that in the negative group (P=0.034, P=0.007, P=0.048, P=0.029). The levels of IL-6 and IL-10 in neonates delivered by vagina were significantly higher than those by cesarean section, while the ratio of Th1/Th2 was lower than that by cesarean section (P<0.001). The level of IL-10 in positive group of neonatal HBsAg was significantly higher than that in negative group, while TNF-α and Th1/Th2 ratio were lower than negative group (P=0.011, P<0.001, P=0.027). The degree of Th2 predominant response was reflected by ratio of Th1/Th2. After adjusting potential confounding factors in non-conditional logistic regression analysis, compared to those born to mothers with PBMC HBV cccDNA negative, neonates whose mother with PBMC HBV cccDNA positive had an increased risk of having a strong Th2 predominant response (OR=2.42,95% CI:1.16-5.04, P=0.018). The risk of a strong Th2 predominant response in neonates delivered by vagina was 5.49 times higher than those by cesarean section (OR=5.06, 95% CI: 2.95-8.67, P<0.001). Conclusion HBsAg-positive mothers’ PBMC HBV replication and vaginal delivery may increase the risk of having a Th2 predominant response in neonates. It is suggested that we should pay attention to the effect of maternal PBMC HBV replication and the mode of delivery on neonatal Th1/Th2 cytokines.

Relationship between Maternal PBMC HBV cccDNA and HBV Serological Markers and its Effect on HBV Intrauterine Transmission / 生物医学与环境科学（英文）

OBJECTIVE@#To investigate the relationship between maternal peripheral blood mononuclear cells (PBMC) hepatitis B virus (HBV) covalenty closed circular deoxyribonucleic acid (cccDNA) and other HBV serological markers and its effects on HBV intrauterine transmission.@*METHODS@#We enrolled 290 newborns and their hepatitis B surface antigen (HBsAg) positive mothers. HBV cccDNA in PBMC and HBV DNA in serum were detected by a real-time PCR-TaqMan probe while HBV serological markers were detected with an electrochemiluminescence immunoassay.@*RESULTS@#There was a positive correlation between the levels of PBMC HBV cccDNA and serum HBV DNA and HBeAg (r = 0.436 and 0.403, P < 0.001). The detection rate of pattern A ['HBsAg (+), HBeAg (+), and anti-HBc (+)'] was significantly higher in the PBMC HBV cccDNA positive group than in the control group (χ2 = 48.48, P < 0.001). There was a significant association between HBV intrauterine transmission and PBMC HBV cccDNA (χ2 = 9.28, P = 0.002). In the presence of serum HBV DNA, HBeAg, and PBMC HBV cccDNA, the risk of HBV intrauterine transmission was three times higher (OR = 3.69, 95% CI: 1.30-10.42) than that observed in their absence. The risk of HBV intrauterine transmission was the greatest (OR = 5.89, 95% CI: 2.35-14.72) when both PBMC HBV cccDNA and pattern A were present. A Bayesian network model showed that maternal PBMC HBV cccDNA was directly related to HBV intrauterine transmission.@*CONCLUSION@#PBMC HBV cccDNA may be a direct risk factor for HBV intrauterine transmission. Our study suggests that serological markers could be combined with PBMC-related markers in prenatal testing.

Relationship between the ende mic arsenism and the liver,renal damage / 中国地方病学杂志

Objective To explore the relationship between the endemic arsenism and the liver,renal damage.Methods Some permanent residents were selected as investigated subjects who lived at 3 villages in Datong in Shanxi Province,an arseniasis-endemic areas,These objects were divided into arsenic poisoning and control group on the basis of Diagnosis Standard for Endemic Arsenism(WS/T 211-2001).Then blood and urine samples were collected in the surveyed people.Serum glutamate pyruvic transaminase(ALT)were detected by Enzyme-linked immunosorbent assay as the indicator of the impaired hepatic function.The microdosis albumen (mAlb)and acetylglucosaminidase(NAG)in urine were detected by end-point method and alkaline picric acid as the renal damage indicators.Results A total of 661 people investigated,of which 144 cases were arsenic poisoning patients.The rates of abnormal liver function were significant hisher in arsenic poisoning group[10.42% (15/144)]than that in control[5.22%(27/517)],and both wag significant[X2=5.107,P＜0.05;OR=2.11,95%CI (1.09-4.08)].The geometric mean of mAlb/Ucr was 2.16 mg/g Cr in control,and 2.31 mg/g Cr in arsenic poisoning group,and both was not significant(t=-1.71,P＞0.05).The geometric mean of NAG waft higher in arsenic poisoning group(2.43 U/g Cr)than that in the control(2.22 U/g Cr),and both was significant(t=-3.55, P＜0.05).Conclusions The damage of the liver and renal function were related with endemic arsenism,and NAG is the early indicators suggesting impaired renal function due to endemic arsenism.

Comprehensive study on the risk factors of hepatitis B virus intrauterine infection / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the risk factors of hepatitis B virus (HBV) intrauterine infection.</p><p><b>METHODS</b>Risk factors of HBV intrauterine infection were analyzed by nested case control study.</p><p><b>RESULTS</b>Data from univariate analysis revealed that risk factors of HBV intrauterine infection were positive results on HLA-DR3 (OR = 4.71, 1.62-13.66), HBV DNA (OR = 6.59, 2.72-15.97) and HBeAg (OR = 4.53, 1.93-10.64) in pregnant women, HLA-DR3 (OR = 3.91, 1.18-12.94) in newborn, HLA-I) R3 (OR = 5.96, 1.14-31.15) both in pregnant women and her newborns and HBV infection in placentas (OR = 2.51,1.12-5.60). Results from Multivariate unconditional logistics regression analysis showed that the risk factors of HBV intrauterine infection were positive in both HLA-DR3 (OR = 4.65, 1.44-15.05) and HBV DNA (OR = 6.56, 2.65-16.23) in pregnant women. However, there was no interaction between the two factors. The exposure rate of other factors did not reveal the difference in the two groups. With the increase of HBV DNA in pregnant women, the risk of HBV intrauterine infection was rising (chi2 = 16.74, P < 0.05).</p><p><b>CONCLUSION</b>Risk factors of HBV intrauterine infection were HLA-DR3 positive and HBV DNA positive in pregnant women but there was no interaction between the two factors. The risk of HBV intrauterine infection was increased along with the increase of HBV DNA in pregnant women.</p>