Erratum: Ophiopogonin-B Suppresses Epithelial-mesenchymal Transition in Human Lung Adenocarcinoma Cells via the Linc00668/miR-432-5p/EMT Axis: Erratum.

[This corrects the article DOI: 10.7150/jca.31338.].

Erratum: Ophiopogonin B inhibits migration and invasion in non-small cell lung cancer cells through enhancing the interaction between Axin and ß-catenin: Erratum.

[This corrects the article DOI: 10.7150/jca.60066.].

IAIH-PG consensus for histological criteria of AIH: Multicentre validation with focus on chronic liver diseases in China.

BACKGROUND AND AIMS: The International AIH Pathology Group (IAIH-PG) put forward the new histological criteria of autoimmune hepatitis (AIH) in 2022, which have not undergone adequate verification. In this study, we verified the applicability of the new histological criteria in the population of Chinese patients with chronic liver disease, comparing it with the simplified criteria. METHODS: The gold standard for diagnosis in all patients was based on histological findings, combined with clinical manifestations and laboratory tests and determined after a follow-up period of at least 3 years. A total of 640 patients with various chronic liver diseases from multiple centres underwent scoring using the new histological criteria and the simplified criteria, comparing their diagnostic performance. RESULTS: In this study, the new histological criteria showed a sensitivity of 73.6% and 100% for likely and possible AIH, with specificities of 100% and 69.0% respectively. The coincidence rates of possible AIH for the new histological criteria, simplified histological criteria and simplified score were 81.7%, 72.8% and 69.7% respectively. For likely AIH, the rates were 89.2%, 75.9% and 65.6% respectively. Based on the new histological criteria, all patients with AIH were correctly diagnosed. Specifically, 73.6% were diagnosed with likely AIH and 26.4% were possible AIH. Additionally, the simplified histological criteria achieved a diagnosis rate of 98.6% for AIH, while the simplified score could only diagnose 53.8% of AIH. CONCLUSIONS: Compared with the simplified score and simplified histological criteria, the sensitivity and specificity of the new histological criteria for AIH were significantly improved. The results indicate that the new histological criteria exhibit high sensitivity and specificity for diagnosing AIH in China.

Development and validation of a noninvasive prediction model of autoimmune hepatitis in patients with liver diseases.

BACKGROUND AND AIMS: There is no golden standard for the diagnosis of autoimmune hepatitis which still dependent on liver biopsy currently. So, we developed a noninvasive prediction model to help optimize the diagnosis of autoimmune hepatitis. METHODS: From January 2017 to December 2019, 1739 patients who had undergone liver biopsy were seen in the second hospital of Nanjing, of which 128 were here for consultation. Clinical, laboratory, and histologic data were obtained retrospectively. Multivariable logistic regression analysis was employed to create a nomogram model that predicting the risk of autoimmune hepatitis. Internal and external validation was both performed to evaluate the model. RESULTS: A total of 1288 patients with liver biopsy were enrolled (1184 from the second hospital of Nanjing, the remaining 104 from other centers). After the univariate and multivariate logistic regression analysis, nine variables including ALT, IgG, ALP/AST, ALB, ANA, AMA, HBsAg, age, and gender were selected to establish the noninvasive prediction model. The nomogram model exhibits good prediction in diagnosing autoimmune hepatitis with AUROC of 0.967 (95% CI: 0.776-0.891) in internal validation and 0.835 (95% CI: 0.752-0.919) in external validation. CONCLUSIONS: ALT, IgG, ALP/AST, ALB, ANA, AMA, HBsAg, age, and gender are predictive factors for the diagnosis of autoimmune hepatitis in patients with unexplained liver diseases. The predictive nomogram model built by the nine predictors achieved good prediction for diagnosing autoimmune hepatitis.

Induction of Ferroptosis by Ophiopogonin-B Through Regulating the Gene Signature AURKA in NSCLC.

Ferroptosis is a new type of iron-dependent programmed cell death. In recent years, its role in the diagnosis and treatment of multiple tumors, including non-small cell lung cancer (NSCLC), has been continuously observed. The relationship between the ferroptosis-related genes and the prognosis of patients with NSCLC needs to be clarified. In this study, The Cancer Genome Atlas (TCGA) and the Gene Expression Synthesis database (Gene Expression Omnibus, GEO) were used to build a model of ferroptosis-related differentially expressed genes (DEGs). A total of 101 ferroptosis-related DEGs were screened using R language, and a 12-gene signature was finally established through univariate Cox regression analysis and least absolute shrinkage and selection operator (LASSO)-penalized Cox regression analysis. According to the risk scores, the patients were divided into a high-risk or a low-risk group, with patients in the low-risk group showing better prognosis. AURKA, one of the genes in the 12-gene signature, was found to be highly expressed in tumors. In addition, further study verified AURKA to be a negative regulator of ferroptosis in NSCLC cells. Ophiopogonin B (OP-B) had been reported to induce apoptosis, mitotic catastrophe, and autophagy in NSCLC cells. Herein, proteomic sequencing analysis and OP-B administration revealed the upregulation of AURKA and the downregulation of PHKG2 and SLC7A5 in the 12-gene signature, indicating that OP-B induced ferroptosis in NSCLC. Determination of the concentrations of malondialdehyde (MDA), glutathione (GSH), and intracellular iron and the mitochondrial membrane potential (MMP) confirmed the induction of ferroptosis by OP-B in vitro. Furthermore, transmission electron microscopy (TEM) examination of lung cancer xenotransplantation in nude mice confirmed that OP-B induced ferroptosis in vivo. Further study of the molecular mechanism showed that the ferroptosis effect caused by OP-B can be partially reversed by the overexpression of AURKA. Overall, our study established a new ferroptosis-related risk prediction model for the prognosis of patients with NSCLC, revealed the enrichment pathways of ferroptosis in NSCLC, and discovered the negative regulation of AURKA in ferroptosis. On this basis, we demonstrated that OP-B can induce ferroptosis in NSCLC and clarified the specific molecular mechanism of OP-B inducing ferroptosis by regulating the expression of AURKA.

Ophiopogonin B alleviates cisplatin resistance of lung cancer cells by inducing Caspase-1/GSDMD dependent pyroptosis.

Drug resistance has become the main reason for the failure of tumor chemotherapy. Radix Ophiopogon Japonicus has long been used as traditional Chinese medicine to treat pulmonary disease, and Ophiopogonin B (OP-B) as a bioactive component of it has also been verified to inhibit cell proliferation of various non-small cell lung cancer (NSCLC) cells in vivo and in vitro. Therefore, we wonder whether OP-B is also effective to drug resistant lung cancer cells. Firstly, Cell Counting Kit-8 (CCK8) assay was used to compare the sensitivity of OP-B on NCI-H460, A549, cisplatin resistant A549 (A549/DDP) and paclitaxel resistant A549 (A549/PTX) cells, and A549/DDP cells were shown to be more sensitive to OP-B than other three cell lines, the results were further verified in orthotopic tumor nude mice model and zebrafish tumor model. Moreover, observation of cell morphological feature, mitochondrial membrane potential, LDH release rate, and production of IL-1ß all suggested that OP-B induced pyroptosis in A549/DDP cells more significantly than that in A549 cells. Meanwhile, transcriptomic sequencing results between OP-B treated and the Mock A549/DDP group also suggested that OP-B induced more significant Caspase-1/GSDMD dependent pyroptosis in A549/DDP group, which was further verified by VX-765, the inhibitor of Caspase-1. Together, the experimental results suggested that OP-B alleviated DDP resistance of A549 cells through inducing more significant Caspase-1/GSDMD-dependent pyroptosis.

Effects of Gentiopicroside on activation of NLRP3 inflammasome in acute gouty arthritis mice induced by MSU.

Acute gouty arthritis is a self-limiting inflammatory disease resulting from the deposition of monosodium urate (MSU) crystals. It has been shown that Gentiopicroside (GPS) possesses anti-inflammatory and analgesic functions. The aim of this study was to parse out whether GPS has an effect on acute gouty arthritis. We established an acute gouty arthritis model by the injection of MSU into the paw, and found that GPS relieves MSU-induced mechanical, thermal hyperalgesia, and paw swelling. Furthermore, GPS down-regulated the release of pro-inflammatory cytokines in paw tissues, including IL-1ß, IL-6, IL-18, and TNF-α. The results of H&E staining and MPO activity measurement showed that GPS inhibits neutrophil infiltration. And the over-expressions of NOD-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), and Caspase-1 induced by MSU were inhibited by treatment with GPS. These results revealed that GPS can treat acute gouty arthritis based on anti-inflammatory and analgesic properties in vivo, which might be ascribed to the inhibition on NLRP3 inflammasome. Furthermore, we performed in vitro study to confirm the results of in vivo study. Consistently, the results proved that GPS could inhibit the activation of NLRP3 inflammasome in RAW264.7 macrophages stimulated by LPS-MSU. In conclusion, this study provides an experimental basis for the application of GPS and expands the potential value of GPS in the therapy of acute gouty arthritis.

Ophiopogonin B inhibits migration and invasion in non-small cell lung cancer cells through enhancing the interaction between Axin and ß-catenin.

Ophiopogonin B (OP-B), a kind of saponin compound that exists in Radix Ophiopogonis is frequently adopted for the treatment of lung disease as traditional Chinese medicine. The present work aimed to explore the anti-tumor activity of OP-B on non-small cell lung carcinoma (NSCLC) and its possible mechanism. We found that OP-B-treated cells suppressed the viability and proliferation of cells depending on its concentration, as assayed by MTT and Alamar Blue (IC50 were 14.22 ± 1.94, 12.14 ± 2.01, and 16.11 ± 1.83 µM in A549, NCI-H1299, and NCI-H460 cells, respectively). Then, the suppressive effect of OP-B on the invasion and migration of NSCLC was observed through wound healing and Transwell assays, and the epithelial-mesenchymal transition (EMT) markers was detected by immunofluorescence and western blotting. In addition, a dose-dependent reduction of ß-catenin both within cytoplasm and nucleus was observed, and the downstream proteins cyclin D1 and c-Myc of Wnt/ß-catenin pathway were also reduced. We further constructed ß-catenin-overexpression cell models to reveal the underlying mechanism. The results showed that 10 µM of OP-B notably reduced ß-catenin protein levels, as well as cell migration and invasion. In spite of the increasement of ß-catenin, activation of Wnt pathway and EMT progression, knockdown of Axin leaded to de-function of OP-B on cell metastasis. Taken together, OP-B reduced NSCLC migration and invasion by strengthening the Axin/ß-catenin interaction and reducing ß-catenin protein translocation.

The effects of the miR-21/SMAD7/TGF-ß pathway on Th17 cell differentiation in COPD.

Chronic obstructive pulmonary disease (COPD) is a complex disease with multiple etiologies, while smoking is the most established one. The present study investigated the modulation of T-helper 17 (Th17) cell differentiation by the miR-21/Smad7/TGF-ß pathway, and their roles in COPD. Lung tissues were obtained from lung cancer patients with or without COPD who underwent lobotomy and the levels of miR-21, TGF-ß/Smad signaling molecules, RORγT, and other Th17-related cytokines were detected. Mouse COPD models were built by exposing both wild-type (WT) and miR-21-/- mice to cigarette smoke (CS) and cigarette smoke extract (CSE) intraperitoneal injection. Isolated primary CD4+ T cells were treated with either CS extract, miR-21 mimics or inhibitors, followed by measuring Th17 cells markers and the expression of TGF-ß/Smad signaling molecules and RORγT. Increased levels of miR-21, Smad7, phosphorylated (p)-Smad2, p-Smad3, TGF-ß, and Th17-related cytokines was detected in the lungs of COPD patients. Lung function in modeled WT mice, but not miR-21-/- ones, deteriorated and the number of inflammatory cells in the lung tissues increased compared to the control WT-mice. Moreover, primary CD4+ lymphocytes tend to differentiate into Th17 cells after the treatment with CSE or miR-21 mimics, and the expression of RORγT and the TGF-ß/Smad signaling were all increased, however miR-21 inhibitors worked reversely. Our findings demonstrated that Th17 cells increased under COPD pathogenesis and was partially modulated by the miR-21/Smad7/TGF-ß pathway.

Study on metastasis inhibition of Kejinyan decoction on lung cancer by affecting tumor microenvironment.

BACKGROUND: Kejinyan decoction, as an experienced formula of Zhou Zhongying (the Master of Traditional Chinese Medicine) has been widely used in clinic for lung cancer treatment in China, while the anti-lung cancer mechanism of it is still remained to be elucidated. Herein, our basic study found that the survival of lung cancer xenograft mice was significantly prolonged after intragastrically administered high dose of Kejinyan decoction (3.8 g per kg BW) for 15 days. More importantly, we found that Kejinyan decoction inhibited the metastasis of lung cancer cells in vivo. Thus in this study, we aim to elucidate the anti-metastasis effects of Kejinyan decoction. METHODS: RNA-Seq was used to find out the gene regulation of Kejinyan decoction on the mice, flow cytometry assay was used to detect the immunocytes in the spleen, ELISA assay was used to detect the inflammatory factors in the serum and spleen, and immunofluorescence assay was used to detect the level of immune cells and the expression of glycol-metabolism related enzymes in situ. Also, we established a lung cancer orthotopic xenograft tumor model to assess the influence of Kejinyan decoction on the metastatic ability of lung cancer cells in vivo. RESULTS: GO analysis of gene sequencing of tumor tissue samples showed that Kejinyan decoction regulated immune response. Further flow cytometry analysis of splenic lymphocyte showed that Kejinyan decoction upregulated M1 macrophages and downregulated M2 macrophages, while the total level of macrophages changed little, which was verified by detection of CD68, F4/80, CD206, and CD86 in tumor tissue section. Moreover, detection of inflammatory cytokines showed that Kejinyan decoction downregulated TNF-α, IFN-γ, IL-6, as well as IL-4, IL-13 in tumor microenvironment. Further studies also showed that Kejinyan decoction had little effect on tumor hypoxia, but downregulated glycolysis in tumor tissues. More importantly, we found that Kejinyan decoction inhibited the metastasis of lung cancer cells in vivo. CONCLUSION: Our findings conclude that Kejinyan decoction inhibited lung cancer cell metastasis through affecting macrophage polarization and energy reprogramming.

Ophiopogonin-B Suppresses Epithelial-mesenchymal Transition in Human Lung Adenocarcinoma Cells via the Linc00668/miR-432-5p/EMT Axis.

Ophiopogonin-B (OP-B) has been reported to suppress metastasis and angiogenesis of adenocarcinoma A549 cells in vitro and in vivo. More and more evidences indicate that inflammatory microenvironment facilitates tumor metastasis. Digital Gene Expression (DGE) analysis of non-small cell lung cancer (NSCLC) cell lines showed that OP-B downregulated the expression of linc00668, which promoted progression of cancer. Herein, we simulated the inflammatory microenvironment by co-culturing A549 cells with LPS-treated THP-1 cells and found that the level of linc00668 increased significantly in the mock group, while OP-B treatment inhibited the level of linc00668 and reversed epithelial-mesenchymal transition (EMT) induced by linc00668. In addition, overexpression of linc00668 in A549 cells suppressed the expression of E-cadherin and induced expression of N-cadherin, while OP-B treatment reversed these changes. Bioinformatic prediction and dual-luciferase reporter gene assay validated that linc00668 sponge miR-432-5p and at last acted on EMT to execute the anti-migration function of A549 cells under inflammatory microenvironment. Taken together, OP-B inhibits metastasis of A549 cells via the linc00668/miR-432-5p/EMT axis.

A Novel Murine Chronic Obstructive Pulmonary Disease Model and the Pathogenic Role of MicroRNA-21.

Chronic obstructive pulmonary disease (COPD) is a multi-pathogenesis chronic lung disease. The mechanisms underlying COPD have not been adequately illustrated. Many reseachers argue that microRNAs (miRs) could play a crucial role in COPD. The classic animal model of COPD is both time consuming and costly. This study proposes a novel mice COPD model and explores the role of miR-21 in COPD. A total of 50 wide-type (WT) C57BL/6 mice were separated into five euqlly-sized groups-(1) control group (CG), (2) the novel combined method group (NCM, cigarette smoke (CS) exposure for 28 days combined with cigarette smoke extract (CSE) intraperitoneal injection), (3) the short-term CS exposure group (SCSE, CS exposure for 28 days), (4) the CSE intraperitoneal injection group (CSEII, 28 days CSE intraperitoneal injection), and (5) the long-term CS exposure group (LCSE, CS exposure).The body weight gain of mice were recorded and lung function tested once the modeling was done. The pathological changes and the inflammation level by hematoxylin eosin (H&E) staining and immunohistochemical staining (IHS) on the lung tissue sections were also evaluated. The level of miR-21 in the mice lungs of the mice across all groups was detected by RT-qPCR and the effects of miR-21 knock-down in modeled mice were observed. The mice in LCSE and NCM exhibited the most severe inflammation levels and pathological and pathophysiological changes; while the changes for the mice in SCSE and CSEII were less, they remained more severe than the mice in the CG. The level of miR-21 was found to be negatively correlated with lung functions. Moreover, knocking miR-21 down from the modeled mice, ameliorated all those tested COPD-related changes. Our novel modeling method detected virtually the same changes as those detected in the classic method in WT mice, but in less time and cost. Further, it was determined that the level of miR-21 in the lungs could be an indicator of COPD severity and blocking functions of miR-21 could be a potential treatment for early stage COPD.

[Progress in study on animal models of chronic obstructive pulmonary disease].

Chronic obstructive pulmonary disease (COPD) is an incompletely reversible chronic airway disease that can be prevented and cured. There is a tendency toward increasing the morbidity and mortality for COPD. Establishment of an animal model for COPD is an important step to explore the pathogenesis of this disease. Presently, a well-recognized COPD animal model is not available. The key points for establishing the COPD animal models, such as selection of animal species, parameters for model evaluation, are constantly updated.

Rapid immunization effects of a new type of 60 µg hepatitis B vaccine compared with traditional 20 µg hepatitis B vaccines in adults.

BACKGROUND: The widely recommended standard schedule of hepatitis B vaccine for adults is months 0, 1 and 6, which takes 6 months to complete. Rapid completion of one vaccination schedule is important to adults because of its low compliance with follow-up doses. A new type of 60 µg Hepatitis B vaccine, made by Shenzhen Kangtai Biological Products Co., LTD., is originally recommended for low or non-responders. The objective of this clinical trial was to test whether this 60 µg hepatitis B vaccine could be used in primary immune population and what is its level of immunogenicity and safety compared with other hepatitis B vaccines. METHODS: This is a 2-center randomized controlled study. A total of 1169 healthy adults aged between 25 and 55 who tested negative for HBsAg, anti-HBs, and anti-HBc were eligible for the study and were enrolled from relatively fixed and stable sites, such as villages, schools and large enterprises et al in Xuanhua county in Hebei province and Huaibei county in Anhui province. They were randomized to group A (20 µg Engerix-B® with 0, 1, 6 month intervals), group B (20 µg Kangtai hepatitis B vaccine with 0, 1, 6 month intervals), group C (60 µg Kangtai hepatitis B vaccine with 0, 2 month intervals) and group D (20 µg Huabei hepatitis B vaccine made by recombinant DNA techniques in CHO cell with 0, 1, 6 month intervals). In group A, B and D, every study object's blood sample was collected in the second month after their final injection to test the anti-HBs levels; while in group C, the blood sample was collected in the second month after the first and the second injection to test the anti-HBs levels. Adverse events were collected after each dose to assess the vaccines' safety. RESULTS: The seroprotection rates were 93.17%, 97.23%, 93.54% and 98.98% respectively and the geometric mean titers (GMTs) were 1033.38 mIU/ml, 600.75 mIU/ml, 265.69 mIU/ml and 1627.05 mIU/ml in group A,B,C and D respectively. The difference of seroprotection rate among the 4 groups was statistically significant (χ2 = 17.26, P<0.05). The difference of titers of anti-HBs among the 4 groups was statistically significant (H = 162.42, P<0.05). BMI, age (older than 40) and smoking were the influence factors of anti-HBs levels on 60 µg hepatitis B vaccine. CONCLUSION: 60 µg hepatitis B vaccine has a satisfactory safety and seroprotection rate in adult, It could be used in rapid adult hepatitis B immunization.

[Survey on seroepidemiological status and vaccine coverage of hepatitis B among children in Chaoyang district of Beijing in 2010].

OBJECTIVE: To explore seroepidemiological status and vaccine coverage of hepatitis B in children aging under 15 years old in Chaoyang district of Beijing. METHODS: A total of 1602 children aging under 15 years old, residents or floating population who had lived here more than six months, were randomly selected by multistage cluster sampling, from Chaoyang district of Beijing in year 2010. The demographic information and vaccine coverage of hepatitis B vaccine (HepB) were collected by self-designed questionnaire.5 ml blood was collected from each subject and the serum HBsAg, anti-HBs and anti-HBc were detected by Abbott microparticle enzyme-linked immunoassay. Those whose HBsAg was positive were then tested HBeAg and anti-HBe. The positive rate of hepatitis B indicators and coverage rate of HepB in different population were compared. RESULTS: The positive rate of HBsAg, anti-HBs and anti-HBc were 0.56% (9/1602), 64.17% (1028/1602) and 2.12% (34/1602), respectively; while the age standardized rates were separately 0.57%, 66.36% and 1.98%; and the gender-adjusted rates were 0.56%, 64.23% and 2.12% respectively. The positive rate of anti-HBs was statistically significant (χ(2) = 165.445, P = 0.000). The positive rate of anti-HBs was up to 90.73% (235/259) among 1-2 years old children, followed by 76.22% (141/185) among 13 - 15 years old children, 67.21% (166/247) among 3 - 4 years old children, 61.22% (150/245) among 9 - 10 years old children, 60.68% (142/234) among 11 - 12 years old children, 49.05% (103/210) among 5 - 6 years old children and 40.99% (91/222) among 7 - 8 years old children. The average coverage rate of HepB was 90.44% (1371/1516), separately 93.76% (661/705) in residents and 87.55% (719/811) in floating population. The difference was statistically significant (χ(2) = 16.829, P = 0.000). CONCLUSION: HBsAg positive rate in children under 15 years old in Chaoyang district of Beijing dropped to less than 1% and the coverage rate of HepB had reached over 90%. It is suggested that we should pay more attention to increase the coverage rate of HepB among floating children under 15 years old.

[Study on family aggregation and risk factors of hepatitis B virus transmission in Chaoyang district, Beijing].

OBJECTIVE: To explore the family aggregation and risk factors of hepatitis B virus (HBV) transmission in Chaoyang district of Beijing. METHODS: A total of 5266 families were randomly selected for the multi-stage cluster sampling study in Chaoyang district of Beijing in 2010. The family members who aged between 1 and 70 years old and lived constantly in Beijing for over half a year, were recruited as subjects. There were 14 491 subjects in total, including temporary residents who did not have Beijing household account, except foreigners. 5 ml venous blood was drawn from every subject. A self-designed questionnaire was used to collect the basic information of the population and the risk factors of the hepatitis B transmission. Microparticle enzyme-linked immunoassay was applied to test five indicators of hepatitis B. Negative binomial distribution test was used among the HBsAg positive families to calculate the family aggregation rate of hepatitis B. Single factor analysis and multi-factor logistic regression model were used to analyze the risk factors of HBV transmission. RESULTS: In all, 308 out of 5266 families had HBsAg positive members, accounting for 5.85%.383 out of 14 410 subjects were HBsAg positive, rating at 2.66%. The HBsAg positive rate among subjects under 14 years old was the lowest, at 0.56% (9/1603); and the positive rate among subjects aging between 35 and 44 years old was the highest, at 4.27% (47/1029). Negative binomial distribution test showed that the family aggregation rate of HBV infection was 7.66% (χ² = 15.10, P < 0.05). The analysis of family aggregation of HBsAg positive showed that 17.39% (8/46) of the transmission was from father to child, 13.04% (6/46) was from mother to child, 30.44% (14/46) was between couples, and another 39.13% (18/46) was between siblings or other relatives. Both single factor analysis and multi-factor logistic regression analysis showed that hepatitis B positive family members (OR = 5.40, 95%CI: 5.24 - 5.55), hepatitis B positive friends and colleagues (OR = 1.55, 95%CI: 1.11 - 1.99) and blood donation and transfusion history (OR = 1.96, 95%CI: 1.76 - 2.15) were the risk factors of HBV infection. CONCLUSION: HBV transmission showed family aggregation in Beijing, however, the risk factors needed further studies.

[Prevalence of hepatitis B in Chaoyang district, Beijing in 2010].

OBJECTIVE: To study the prevalence of hepatitis B infections and carrier status among general population in Chaoyang district, Beijing in 2010. METHODS: From May to December 2010, 14 491 subjects over 12 months old were selected by multistage random cluster sampling method from residents in Chaoyang district, Beijing. Five millilitre venous blood specimens were collected from these subjects to test hepatitis B virus antigens and antibodies. Status of hepatitis B infections were analyzed in different age, sex and registered permanent residence groups. RESULTS: The overall positive rate of surface antigen (HBsAg) was 2.66% (383/14 410). The lowest rate of 0.56% (9/1603) was found in the 1 to 14 years old group and the 35 to 44 years old group had the highest rate of 4.27% (92/2154). The rate in subjects younger than 24 years old was 1.03% (31/2986). The overall positive rate of surface antibody (anti-HBs) was 40.21% (5798/14 421). The highest positive rate of anti-HBs (80.59%, 407/505) was found in the 1 to 4 years old group. The overall positive rate of core antibody (anti-HBc) was 30.26% (4364/14 424). The overall hepatitis B virus infection rate was 30.32% (4364/14 393). For male and female groups, the positive rates of HBsAg were 2.93% (179/6108) and 2.44% (202/8287) respectively (χ² = 3.32, P > 0.05); anti-HBs were 41.93% (2563/6113) and 38.96% (3231/8293) respectively (χ² = 12.88, P < 0.01); and anti-HBc were 31.39% (1919/6114) and 29.39% (2438/8295) respectively (χ² = 6.65, P = 0.01). For local residents group and mobile population group, the positive rates of HBsAg were 2.46% (283/11 510) and 3.60% (98/2719) respectively (χ² = 11.08, P < 0.01); anti-HBs were 37.11% (4293/11 568) and 53.07% (1445/2723) respectively (χ² = 233.51, P < 0.01); and anti-HBc were 30.83% (3567/11 570), and 28.41% (774/2724) respectively (χ² = 6.08, P < 0.05). CONCLUSION: The positive rate of HBsAg in population younger than 24 years old has reached a relatively low level. The mobile population has significantly higher positive rate of HBsAg than local residents, indicating the need for enhancing prevention and control measures for hepatitis B for the mobile population and local residents over 25 years old.

[Comparison on the antibody response after primary immunization of 5 µg and 10 µg hepatitis B vaccine made by recombinant DNA techniques among newborns].

OBJECTIVE: To compare the antibody response induced by primary immunization with 5 µg and 10 µg hepatitis B vaccine made by recombinant DNA techniques among the newborns. METHODS: Healthy infants who had completed primary immunization with 5 µg hepatitis B vaccine made by recombinant dexyribonucleic acid techniques in Saccharomyces (Hep-SC) or 10 µg hepatitis B vaccine made by recombinant dexyribonucleic acid techniques in Hansenula polymorpha (HepB-HP) were included in the study. Kids under study were 7-12 months of age and had been on 0-1-6 schedule. Standardized questionnaire was used and blood samples were collected. The titer of antibody to hepatitis B surface antigen (anti-HBs) was detected by Chemiluminescence Microparticle Imunoassay (CMIA). If anti-HBs happened to be under 10 mIU/ml, HBV DNA was further detected by nested-PCR to distinguish occult hepatitis B virus infection. Sero-conversion rate and titer of anti-HBs were compared between the two kinds of hepatitis B vaccines. Multivariate analysis was used to find the relationship between the kind of hepatitis B vaccine as well as the antibody response after debugging the other influencing factors including month-age, gender, birth-weight, premature birth and mother's HBsAg status. RESULTS: 8947 infants vaccinated with 5 µg HepB-SC and 4576 infants vaccinated with 10 µg HepB-HP were investigated. In the 5 µg group, the rates of non-, low-, normal- and high-response were 1.88%, 15.18%, 61.42% and 21.52% respectively. In the 10 µg group, the corresponding rates were 0.15%, 2.16%, 29.42% and 68.26% respectively. The non-, low-, normal-response rates were all higher in 5 µg group than in 10 µg group (P<0.01), while the high-response rate was much higher in 10 µg group than in 5 µg group (P<0.01). The geometric mean concentration (GMC) of anti-HBs were 354.81 mIU/ml (95%CI: 338.84-363.08 mIU/ml) and 1778.28 mIU/ml (95%CI: 1698.24-1819.70 mIU/ml) in the 5 µg group and 10 µg group respectively. The GMC was statistically higher in the 10 µg group than in the 5 µg group (P<0.001). The sero-conversion rate and GMC were significantly different between the two groups even after debugging the other influencing factors. CONCLUSION: Better anti-HBs response could be achieved by primary immunization with 10 µg HepB-HP than with 5 µg HepB-SC among newborns.

[Multi-center matching study on antibody response between preterm and full-term infants after primary immunization of hepatitis B vaccine].

OBJECTIVE: To compare the antibody response between preterm and full-term infants after primary immunization of hepatitis B vaccine (HepB). METHODS: Infants who were aged 7 - 12 months and had completed primary immunization with 5 µg HepB made by recombinant deoxyribonucleic acid techniques in saccharomyces cerevisiae (HepB-SC) or 10 µg HepB made by recombinant deoxyribonucleic acid techniques in Hansenula polymorpha (HepB-HP) on 0-1-6 schedule were investigated in four provinces (municipality) including Beijing, Shandong, Jiangsu and Guangxi of China. Among them, all preterm infants were selected to form the preterm group and the 1:1 matching full-term infants with the same month-age, gender and residence were randomly selected to form the full-term group. Their HepB history was determined by immunization certificate and all of their parents were interviewed with standard questionnaire to get their birth information. Blood samples were obtained from all anticipants and were tested for Anti-HBs by chemiluminescence microparticle immuno-assay (CMIA). RESULTS: Total anticipants were 648 pairs of infants. The rates of non-response, low-response, normal-response and high-response after the primary immunization were 1.39%, 8.64%, 45.83% and 44.14% in the preterm group, respectively. The corresponding rates were 1.08%, 9.26%, 44.91% and 44.75% in the full-term group. The above four rates did not show significant differences between the two groups (P > 0.05). The geometric mean concentrations (GMC) of anti-HBs in the pre-term and full-term group were 755.14 and 799.47 mIU/ml respectively. There was no significantly difference in the GMCs between the two groups (P > 0.05). Results from multivariable conditional logistic analysis showed that preterm was not an influencing factor to the antibody response after HepB primary immunization among newborns even after debugging the other influencing factors. CONCLUSION: The antibody response after HepB primary immunization were similar among the preterm and full-term infants. The preterm newborns could be immunized under the same HepB immunization strategy.

Surface antibody and cytokine response to recombinant Chinese hamster ovary cell (CHO) hepatitis B vaccine.

OBJECTIVE: To compare the immune responses of the 10 µg and 20 µg doses of CHO hepatitis B vaccine on adults. METHODS: Adults aged 18-45 years who gave a history of never having received hepatitis B vaccine and lacked serologic evidence of infection to hepatitis B virus (HBV) infection or previous vaccination were enrolled into the study. A total of 642 eligible participants were randomized to receive 3 doses of either the 10 µg or the 20 µg formulation of CHO hepatitis B vaccine in a 0-1-6 month schedule. Each study subject had a serologic specimen collected one month following the third vaccine dose that was tested for markers of HBV infection and anti-HBs by Abbott I2000. Persons who tested negative for anti-HBs negative persons were tested for HBV DNA. Logistic regression was used to identify factors associated with antibody response. Among the participants, 153 subjects had their lymphocytes cultivated and tested for cytokine production. Enzyme-linked immunospot (ELISPOT) was used to test spot numbers of IL-4, IFN-γ which produced by lymphocyte. RESULTS: The anti-HBs seroconversion rate was 88.8% (95% CI: 85.4-92.2%) and 95.3% (95% CI: 93.0-97.6%), respectively in 10 µg and 20 µg group. Geometric mean titers (GMT) were 173.42 mIU/ml and 585.51 mIU/ml, respectively in 10 µg and 20 µg groups. Multivariate analysis demonstrated that diabetes, spouse is hepatitis B virus infector, older age and receipt of the 10 µg dose were all negatively associated with antibody response (P<.05). Cellular immunity results showed: IL-4 immunity spot numbers in 20 µg group was higher than 10 µg group. With anti-HBs increased, the IL-4 immunity spot numbers increased significantly which had significant positive correlation (Spearman coefficient=0.538, P<0.0001). IFN-γ spot numbers had no statistical significant between the two groups. CONCLUSION: The humoral immunity and cytokines response among the group that received the 20 µg CHO hepatitis B vaccine dose was superior compared to the group that received the 10 µg dose. The 20 µg dose of CHO hepatitis B vaccine should be prioritized for adult vaccination programs in China.