[Analysis of efficacy and factors influencing sequential combination therapy with tenofovir alafenamide fumarate after treatment with entecavir in chronic hepatitis B patients with low-level viremia].

Objective: To observe the efficacy and factors influencing sequential or combined tenofovir alafenamide fumarate (TAF) after treatment with entecavir (ETV) in patients with chronic hepatitis B (CHB) with low-level viremia (LLV). Methods: 126 CHB cases treated with ETV antiviral therapy in the Department of Infectious Diseases of the First Affiliated Hospital of Nanchang University from January 2020-September 2022 were retrospectively collected. Patients were divided into a complete virologic response (CVR) group (n = 84) and a low-level viremia (LLV) group (n = 42) according to the HBV DNA level during treatment. Clinical characteristics and laboratory indicators of the two groups at baseline and 48 weeks were analyzed by univariate analysis. Patients in the LLV group were divided into three groups according to their continued antiviral treatment regimen until 96 weeks: continued use of ETV as a control group; replacement of TAF as a sequential group; and combination of ETV and TAF as a combined group. The data of the three groups of patients were analyzed by one-way analysis of variance for 48 weeks. HBV DNA negative conversion rate, HBeAg negative conversion rate, alanine aminotransferase (ALT), creatinine (Cr), and liver stiffness test (LSM) were compared among the three groups after 96 weeks of antiviral treatment. Multivariate logistic regression was used to analyze the independent factors influencing the occurrence of HBV DNA non-negative conversion in LLV patients at 96 weeks. Receiver operating characteristic curve (ROC) was used to evaluate the effectiveness of predicting the occurrence of HBV DNA non-negative conversion in LLV patients at 96 weeks. Kaplan-Meier was used to analyze the cumulative negative rate of DNA in LLV patients, and the Log-Rank test was used for comparison. HBV DNA and HBV DNA negative conversion rates during treatment were observed dynamically. Results: Univariate analysis showed statistically significant differences in age, BMI, HBeAg positivity rate, HBV DNA, HBsAg, ALT, AST, and LSM at baseline between the CVR group and the LLV group (P < 0.05). Univariate analysis of variance revealed no statistically significant difference among the three groups of LLV patients at 48 weeks (P > 0.05). HBV-DNA negative conversion rate in the sequential group and the combination group was significantly higher than that in the control group after 96 weeks of treatment (88.89% vs. 41.18%, 85.71% vs. 41.18%, χ (2) = 10.404, P = 0.006). HBeAg negative conversion rate was higher than that of the control group, with no statistically significant difference (P > 0.05).Compared with the control group, ALT, Cr, and LSM in the sequential group and the combined group were equally improved to varying degrees, with a statistically significant difference (P < 0.05). Subsequent use of ETV and HBV DNA at 48 weeks were independent risk factors for HBV DNA positivity at 96 weeks in LLV patients (P < 0.05). The AUC of HBV DNA at 48 weeks was 0.735 (95%CI: 0.578 ~ 0.891), the cut-off value was 2.63 log(10) IU/ml, and the sensitivity and specificity were 76.90% and 72.40%, respectively. DNA conversion rate was significantly lower in LLV patients receiving 48-week ETV and 48-week HBV DNA≥2.63 log10 IU/mL than in patients receiving sequential or combined TAF and 48-week HBV DNA < 2.63 log(10) IU/mL. HBV DNA negative conversion rates in the sequential group and combined group at 72 weeks, 84 weeks, and 96 weeks were higher than those in the control group during the period from 48 weeks to 96 weeks of continuous treatment, and the differences were statistically significant (P < 0.05). Conclusion: Sequential or combined TAF antiviral therapy could more effectively improve the 96-week CVR rate, as well as hepatic and renal function, and alleviate the degree of hepatic fibrosis in CHB patients with LLV following ETV treatment. Subsequent use of ETV and HBV DNA load at 48 weeks were independent predictors of HBV DNA positivity at 96 weeks in LLV patients.

[Clinical value of arterial stiffness assessment on risk prediction of vascular stiffness in the octogenarian elderly].

Objective: This study aimed to analyze clinical factors related to arterial stiffening and establish a risk prediction nomogram of arterial stiffening in the octogenarian(≥80 years). Methods: This study was a retrospective cross-sectional study, which enrolled the octogenarian elderly who underwent physical examination and secondary prevention intervention in the outpatient department of Chinese People's Liberation Army General Hospital from April 2022 to August 2022. Clinical data including demographics, biochemical indicators and medical history were collected. Brachial-ankle pulse wave velocity (baPWV) was detected during the clinical visit. Participants were divided into the control group (baPWV≤1 800 cm/s) and vascular sclerosis group (baPWV>1 800 cm/s). The risk factors of arterial stiffness were analyzed by univariate and logistic regression analysis, and the nomogram model was constructed by R programming language. The predictive effect of the nomogram model was evaluated by the receiver operating characteristic curve (ROC). Results: The median age of the 525 participants was 87.0 (82.0, 92.0) years, 504 (96.0%) were male, 82 in the control group, 443 in the vascular sclerosis group. The baPWV, age, systolic blood pressure, mean arterial pressure and diastolic blood pressure were significantly lower in the control group than those in the vascular sclerosis group (all P<0.05). Logistic regression analysis showed that high-density lipoprotein cholesterol, alanine aminotransferase and amylase were protective factors, and alkaline phosphatase and creatinine were risk factors of arterial stiffening (all P<0.05). The combined nomogram model scores including age, mean arterial pressure and the above five laboratory indicators indicated that mean arterial pressure and serum creatinine levels were strongly correlated with vascular sclerosis. The ROC curve suggested that the nomogram model had good prediction ability. Conclusions: Age, mean arterial pressure, high-density lipoprotein cholesterol, alanine aminotransferase, alkaline phosphatase, amylase and creatinine are independently determinants for increased vascular stiffness. The combined prediction model in this study can provide reference for individualized clinical risk prediction of vascular sclerosis in the octogenarian elderly.

[Heritability and genetic correlation of body mass index and coronary heart disease in Chinese adult twins].

Objective: To examine the heritability of body mass index (BMI) and coronary heart disease (CHD), and to explore whether genetic factors can explain their correlation. Methods: Participants were from 11 provinces/municipalities reqistered in the Chinese National Twin Registry (CNTR) from 2010 to 2018. Participants data were collected from face-to-face questionnaire survey. Bivariate structure equation model was used to estimate the heritability and the genetic correlation of BMI and CHD. Results: A total of 20 340 pairs of same-sex twins aged ≥25 years were included in this study. After adjusting for age and gender, the heritability of BMI and CHD was 0.52 (95%CI: 0.49-0.55) and 0.76 (95%CI: 0.69-0.81), respectively. Further, a genetic correlation was identified between BMI and CHD (rA=0.10, 95%CI:0.02-0.17). Conclusion: In Chinese adult twin population, BMI and CHD are affected by genetic factors, and their correlation can be attributed to the common genetic basis.

Effects of lipids and surfactants on the fermentation production of echinocandin B by Aspergillus nidulans.

AIMS: Echinocandin B (ECB) is a kind of lipopeptide antifungal antibiotic, as well as the key precursor of antifungal drug Anidulafungin. Its efficient bioproduction plays an important role in promoting the industrial production of Anidulafungin. METHODS AND RESULTS: In this study, methyl oleate and Tween 80 were firstly used to enhance the ECB fermentation by Aspergillus nidulans, the results showed that the ECB titre was significantly enhanced with the addition of methyl oleate and Tween 80. Among the lipids, methyl oleate was found to play a pivotal role in increasing the ECB titre to 2123 mg l-1 , which was more than five times higher than that of the control. The addition of Tween 80 in the medium resulted in ECB titre increased to 2584 mg l-1 . The scanning electron microscope (SEM) and N-phenyl-1-naphthylamine (NPN) assay indicated that Tween 80 could influence the cell membrane permeability of A. nidulans, and enhance the intracellular and extracellular substance exchange, therefore lead to the increasing of ECB titre. CONCLUSIONS: Methyl oleate and Tween 80 are optimal carbon sources and surfactants for efficient ECB biosynthesis respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: Surfactant was used in ECB fermentation for the first time, which provided feasible ideas for optimizing the fermentation process of other fungi.

[Effectiveness and safety of ultrasound-guided microwave ablation for the treatment of primary hyperparathyroidism in 12 patients with parathyroid adenoma].

To investigate the effectiveness and safety of ultrasound-guided microwave ablation (MWA) in treatment of primary hyperparathyroidism (PHPT). A total of 12 PHPT patients with parathyroid adenoma were treated with MWA in Nanjing University of Chinese Medicine Affiliated Hospital of Integrated Traditional Chinese and Western Medicine from May 2019 to February 2021. The patients were followed up once every 3 months for 3-12 months. Levels of serum parathyroid hormone (PTH), calcium and phosphorus were detected before and 20 min, 4h and 1day after ablation, and during follow-up period. The volume and volume reduction rate of parathyroid lesion were compared before the treatment and at the end of follow-up. The technical and clinical success of MWA were assessed as well. At the end of follow-up, median serum PTH [66.60 (42.21,80.03) ng/L vs.169.90 (89.01,396.50) ng/L] and calcium [2.39 (2.32,2.49) mmol/L vs. 2.75 (2.57,2.96) mmol/L] levels in 12 patients decreased significantly (all P<0.05). A complete response in terms of PTH and calcium levels was achieved in 6 of the 12 patients, while 4 of the patients had slightly elevated PTH levels just above the upper limit of normal reference range, and 2 of the patients remained abnormal PTH and calcium levels. The clinical cure rate was 50%. The volumes of all lesion after ablation were significantly decreased (P<0.05), with the technical success rate reaching 92.3%. No serious complications were observed. Ultrasound-guided MWA, thus, is safe and effective in the treatment of PHPT.

[Correlation between fasting plasma glucose, HbA1c and DNA methylation in adult twins].

OBJECTIVE: To explore the cytidine-phosphate-guanosine (CPG) sites associated with fas-ting plasma glucose (FPG) and glycated haemoglobin (HbA1c) in twins. METHODS: In the study, 169 pairs of monozygotic twins were recruited in Qingdao, Zhejiang, Jiangsu, Sichuan and Heilongjiang in June to December of 2013 and June 2017 to October 2018. The methylation was detected by Illumina Infinium HumanMethylation450 BeadChip and Illumina Infinium MethylationEPIC BeadChip. According to the Linear Mixed Effect model (LME model), fasting plasma glucose and HbA1c were taken as the main effects, the methylation level (ß value) was taken as the dependent variable, continuous variables, such as age, body mass index (BMI), blood pressure, components of blood cells, surrogate variables generated by SVA, and categorical variables, such as gender, smoking and drinking status, hypoglycemic drugs taking, were included in the fixed effect model as covariates, and the identity numbers (ID) of the twins was included in the random effect model. The intercept was set as a random. Regression analysis was carried out to find out the CpG sites related to fasting blood glucose or HbA1c, respectively. RESULTS: In this study, 338 monozygotic twins (169 pairs) were included, with 412 459 CpG loci. Among them, 114 pairs were male, and 55 pairs were female, with an average age of (48.2±11.9) years. After adjustment of age, gender, BMI, blood pressure, smoking, drinking, blood cell composition, and other covariates, and multiple comparison test, 7 CpG sites (cg19693031, cg01538969, cg08501915, cg04816311, ch.8.1820050F, cg06721411, cg26608667) were found related to fasting blood glucose, 3 of which (cg08501915, ch.8.1820050f, cg26608667) were the newly found sites in this study; whereas 10 CpG sites (cg19693031, cg04816311, cg01538969, cg01339781, cg01676795, cg24667115, cg09029192, cg20697417, ch.4.1528651F, cg16097041) were found related to HbA1c, and 4 of which(cg01339781, cg24667115, cg20697417, and ch.4.1528651f) were new. We found that cg19693031 in TXNIP gene was the lowest P-value site in the association analysis between DNA methylation and fas-ting plasma glucose and HbA1c (PFPG=2.42×10-19, FDRFPG<0.001; PHbA1c=1.72×10-19, FDRHbA1c<0.001). CONCLUSION: In this twin study, we found new CpG sites related to fasting blood glucose and HbA1c, and provided some clues that partly revealed the potential mechanism of blood glucose metabolism in terms of DNA methylation, but it needed further verification in external larger samples.

[Correlation between blood pressure and DNA methylation in adult twins].

OBJECTIVE: To explore the DNA methylation sites correlated with blood pressure (systolic blood pressure, diastolic blood pressure, mean arterial pressure, pulse pressure) in adult twin population. METHODS: A total of 476 twins from the Chinese National Twin Registry were selected as the research population. Questionnaires were used to collect demographic characteristics, lifestyle, disease status and other information, and blood pressure, height, weight and other anthropometric indicators were measured. The genome-wide DNA methylation of whole blood samples was detected by using Infinium HumanMethylation450 BeadChip. The DNA methylation sites correlated with blood pressure were analyzed by constructing mixed effect model with adjusting potential confounding factors, and the significant level was false discovery rate <0.05. RESULTS: After data quality control, 465 twins (122 pairs of monozygotic twins, 104 pairs of dizygotic twins, 13 individuals from 13 pairs of twins) aged (44.8±13.2) years were finally enrolled. There were more males and more monozygotic twins, and the current smokers and current regular drinkers both accounted for more than 30%. No significant CpG site was found after multiple testing in the correlation study between genome-wide DNA methylation and blood pressure by using the collected twins. However, the cg07761116 located on chromosome 10 had low P value in the correlation analysis of 3 blood pressure indices (systolic blood pressure, diastolic blood pressure, mean arterial pressure), suggesting that this site might be correlated with blood pressure. The other 7 sites had low P value in the correlation analysis of the two blood pressure indices, respectively, which pointed to genes involved in neurological development, protein homeostasis, inflammatory reaction and other pathways. CONCLUSION: There is no sufficient evidence to support any DNA methylation site correlated with blood pressure, which may be caused by insufficient sample size and other reasons. This study could provide a reference for subsequent similar twin studies, and subsequent studies can focus on the cg07761116 located on chromosome 10 and other sites with low P values.

[Study on spectrum of UGT1A1 mutations in connection with inherited non-hemolytic unconjugated hyperbilirubinemia].

Objective: To compare and analyze patient's general condition, changes in laboratory parameters, and the spectrum of UGT1A1 mutations in patients with inherited non-hemolytic unconjugated hyperbilirubinemia. Methods: A retrospective study was conducted at Nanjing Second Hospital from January 2015 to July 2018 and patients' demographic characteristics, liver function test, and UGT1A1 gene were analyzed. The categorical variable data were compared by χ (2) test. The normal distribution continuous variable data were compared by t-test and the non-normal distribution continuous variable data were compared using Mann-Whitney U test. Results: Of the 51 patients with inherited non-hemolytic unconjugated hyperbilirubinemia, 44 (86.3%) were Gilbert's syndrome (GS) and seven (13.7%) were Crigler-Najjar syndrome type II (CNS- II). The male to female ratio was 2.9:1 and the average age was 36.11 ± 13.17 years. Six variant types were detected: C. -40_-39insTA, C. -3279T > G, c.211G > A (p.G71R), c.686C > A (p.P229Q), c.1091C > T (p.P364L), c.1456T > G (P.Y486D). Among them, c.211G > A accounted for 58.82% (30/51), c.-40_-39insTA accounted for 27.5% (14/51), and c.1456T > G accounted for 25.5% (13/51). The total bilirubin(TB) and unconjugated bilirubin (UCB) in CNS-II patients were significantly higher than GS patients[155.91 (130 ~ 207) vs. 38.25(29 ~ 52.15) µmol/L, U = 0, P < 0.01; 144.13 (120.8 ~ 197) vs. 30.00 (21.7 ~ 46.75) µmol/L, U = 0.00, P < 0.01, respectively]. Exon mutations of c.1091C > T and c.1456T > G were statistically significant(P < 0.01).There were no differences in age, TB, UCB, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) between the c.211G > A homozygous variants and heterozygous variants (P > 0.05). Conclusion: The common pathogenic mutations of UGT1A1 gene were c.211G > A, c.-40_-39insTA, c.1456T > G. c.211G > A. The mutation has little effect on the level of total bilirubin, but c.1091C > T, c.1456T > G mutations has great influence on the level of total bilirubin.

Development and characterization of microsatellite markers for Solen grandis using Illumina sequencing approach.

Solen grandis is an important economic and overexploited bivalve species. In order to perform its fine-scale genetic analyses, 105 pairs of microsatellites with polymorphism were identified through Illumina Hiseq platform and bioinformatic assembly technology in this study. The estimated fragment size ranged from 100 to 268 bp and the number of alleles per locus varied between 2 and 23. Observed and expected heterozygosities varied from 0.0667 to 1.0000 and 0.0966 to 0.9492, respectively. Fourteen loci deviated significantly from Hardy-Weinberg equilibrium after Bonferroni correction. These microsatellite markers developed in this study would be helpful for future genetic studies on S. grandis and closely related species.

[Evaluating the efficacy of fractional exhaled nitric oxide and impulse oscillometry in screening out cough variant asthma from patients with subacute cough].

Objective: To evaluate the clinical value of fractional exhaled nitric oxide (FeNO) and impulse oscillometry (IOS) in screening out cough variant asthma (CVA) from patients with subacute cough. Methods: Patients with subacute cough were included from the outpatient department of Respiratory Medicine of Zhujiang Hospital of Southern Medical University from May to October in 2016. Based on "the guidelines for the diagnosis and treatment of cough (2015 edition)" , patients were classified into CVA group, and non CVP group with other causes of subacute cough. Lung function, bronchial provocation test, FeNO and IOS were measured. The diagnostic efficiency and optimal cut-off points of FeNO and IOS indicators to diagnose CVA from subacute cough were respectively assessed by the receiver operating characteristic (ROC) curves. Results: A total of 85 patients with subacute cough were included. Among them, 35 patients were diagnosed with CVA (CVA group), the others are classified as non CVP group (n=50). In CVA group, the levels of FeNO and total respiratory impedance (Zrs) were significantly higher, while maximal mid expiratory flow (MMEF)%pred, and mid expiratory flow (MEF)75/50/25%pred, reactance at 5 Hz (X5) levels were significantly lower than those in non CVP group (all P<0.05). Furthermore, the FeNO had a positive correlation with Zrs and Fres (ρ=0.312, P=0.003 and ρ=0.318, P=0.003, respectively), had a negative correlation with X5 (ρ=-0.288, P=0.007). A ROC analysis indicated that the area under ROC curve (AUC) of FeNO in diagnosis of CVA was 0.786 (95% CI: 0.684-0.889), the best cut-off point of FeNO volume ratio was 24.5×10(-9). When FeNO volume ratio=24.5×10(-9,) the sensitivity of in diagnosing CVA was 77.8%, specificity was 70.0%. The AUC for Zrs and X5 were 0.679 and 0.687, respectively. The combination of FeNO and X5 had a greater AUC than other indicators (AUC: 0.817, 95% CI: 0.726-0.908), the sensitivity and specificity were 80.6% and 66.0%, respectively. Conclusion: Both FeNO level and IOS index can be used to screen CVA in patients with subacute cough, and the combination of both have better value in diagnosing CVA.

[Mendelian randomization analysis of the relationship between obesity and DNA methylation].

Objective: To explore the association between DNA methylation and body mass index (BMI) using Mendelian randomization analysis. Methods: A total of 469 participants were selected from the Chinese National Twin Registry in 2013, who were living in Shandong, Jiangsu, Zhejiang, and Sichuan provinces, and at least 18 years of age. A questionnaire survey and physical examination were conducted to collect demographic, clinical, and behavioral information. Peripheral blood cells were collected to detect genotype and methylation status. Association analyses between DNA methylation and BMI and between CpGs and cis-SNP were conducted. With rs748212 as the instrumental variable, the association between cg15053022 and BMI was explored using the Mendelian randomization method. Results: A total of 469 participants were selected. The mean age of participants was (44.8±13.2) years and the BMI was (25.0±3.8) kg/m(2). Nine BMI-related DNA methylation sites were found and DNA methylation site cg15053022 in the ATP4A gene was negatively associated with cis-SNP rs748212 (ß=-0.020); the mean methylation level of AA, AC, and CC were 0.212±0.025, 0.242±0.024, and 0.264±0.028, respectively. rs748212 was associated with BMI (ß=0.04, P=0.007) and closely related to cg15053022 (F=237.66, P=0.143). Mendelian randomization analysis showed lower methylation levels at cg15053022 were associated with higher BMI (ß=-1.97, P<0.001). Conclusion: This study supported the impact of cg15053022 methylation in the ATP4A gene on BMI using Mendelian randomization analysis and provided the basis for using Mendelian randomization analysis in methylation studies.

[Prevalence and risk factors of hypertension among residents of 30 years or above years old in Pengzhou city of Sichuan province].

OBJECTIVE: To investigate the prevalence and risk factors of hypertension among residents in Pengzhou city of Sichuan province. METHODS: Among 20 townships in Pengzhou city of Sichuan Province, 14 townships were selected using a clustered sampling based on the economic level and distance. The registered residents in the selected townships aged 30-79 years were recruited for a comprehensive survey from 2004 to 2007. A total of 55 687 residents (21 315 males and 34 372 females) were selected based on a face to face questionnaire interview and physical examination; the patient group included 14 151 residents with hypertension and the control group had 41 536 residents without hypertension were selected. Multivariate logistic regression model was used to investigate the influence factors for hypertension. OR (95%CI) values were calculated. Different levels of risk exposure factors were transformed into the risk scores using a statistical model. RESULTS: The prevalence rate of hypertension was 25.41% among residents aged 30 years or above in Pengzhou (14 151/55 687), 28.95% (6 170/21 315) for male residents and 23.22% (7 981/34 372) for female residents. Multivaraite logistic regression analysis showed that main risk factors for hypertension in male residents were age, low educational level, widower, low household income, no medical insurance, no fresh fruits intake, high body mass index (BMI), high waist to hip ratio, and low physical activity, which was less than 3 metabolic equivalent of task (MET) level, among which the highest three riskswere (not including the social demographic characteristics) BMI≥28 kg/m(2), waist to hip ratio ≥0.90, and not consuming fresh fruits. OR (95% CI) values for these three key factors were 3.74 (3.27-4.28), 1.34 (1.24-1.44), and 1.27 (1.14-1.41), respectively. The corresponding risk scores for these factors were 2.993, 1.102, and 1.237, respectively. Main risk factors for hypertension in female residents were age, working hours, no medical insurance, fewer number of family members, no fresh fruits intake, smoking, never drinking tea, no dairy products intake, high BMI, highwaist to hip ratio and low physical activity, among which the highest three risks were (not including the social demographic characteristics) BMI ≥28 kg/m(2), MET level, and waist to hip ratio ≥0.85. OR (95% CI) values for these three key factors were 3.30 (3.02-3.60), 1.35 (1.22-1.51), and 1.33 (1.23-1.44), respectively. The corresponding risk scores for these factors were 2.390, 1.263, and 1.051, respectively. CONCLUSION: The prevalence rate of hypertension among the residents over 30 years of age in Pengzhou city was 25.41%. The main risk factors were age, not consuming fresh fruits, high BMI, high waist to hip ratio, and low MET level.

[Role and clinical significance of Th17/Treg balance in patients with severe exacerbation of hepatitis B].

OBJECTIVE: To investigate the role of Th17/Treg balance in immune mechanism in severe exacerbation of hepatitis B. METHODS: The clinical data of 41 patients with chronic hepatitis B were collected, and according to the conditions during hospitalization, these patients were divided into exacerbation group (19 patients) and improvement group (22 patients). On admission, at weeks 1 and 2 of treatment, and at the end of treatment, flow cytometry was used to measure the frequencies of Th17 and Treg cells in peripheral blood, and enzyme-linked immunosorbent assay was used to determine the serum levels of interleukin-17 (IL-17), interleukin-10 (IL-10), and transforming growth factor-ß (TGF-ß). The dynamic changes in the frequencies of Th17 and Treg cells were compared between the two groups, and the correlation between clinical indices for hepatitis and cytokines was analyzed. The t-test was used for comparison between groups, a one-way analysis of variance was used for comparison within one group across different time points, and Pearson correlation analysis was performed. RESULTS: With disease progression, the exacerbation group showed an increase in the frequency of Th17 cells and a relatively low frequency of Treg cells; compared with the improvement group, the exacerbation group had a higher frequency of Th17 cells and a lower frequency of Treg cells. Th17/Treg ratio gradually increased with exacerbation and decreased with improvement in conditions; in the exacerbation group and the improvement group, Th17/Treg ratio was positively correlated with total bilirubin and negatively correlated with prothrombin activity. In the exacerbation group and the improvement group, Th17 cells were positively correlated with IL-17, and Treg cells were positively correlated with IL-10 and TGF-ß. CONCLUSION: Th17 and Treg cells play important roles in severe exacerbation of hepatitis B, and Th17/Treg ratio may be used as an immunobiological marker for the judgment of severity during severe exacerbation of hepatitis B.

Polymorphic microsatellite loci for the razor clam, Sinonovacula constricta.

The razor clam, Sinonovacula constricta, is an important commercial bivalve and a popular mollusca food in China. Twelve polymorphic microsatellite markers were isolated from the razor clam using a partial genomic library enriched for tandem repeat sequences of (CA)16, (GA)16. Polymorphisms of these loci were evaluated in a wild population of 30 individuals. The allele number of these polymorphic markers ranged from 5-15 per locus with an average of 9.333. Observed and expected heterozygosity values ranged from 0.192-1.000 and 0.219-0.906. Polymorphism information content ranged from 0.209-0.892 with an average of 0.704. Three loci significantly deviated from Hardy-Weinberg equilibrium after Bonferroni correction. No significant linkage disequilibrium was detected between these loci. This set of microsatellite loci are useful for genetic studies in S. constricta.

Reference intervals of bone turnover markers determined by using their curve-fitting valley for adult females in China.

SUMMARY: The reference values for bone turnover markers (BTMs) have a significant role in the diagnosis, monitoring, and treatment of metabolic bone disease. This study proposes that the peak value of bone mineral density and the trough value for the BTM curve can be used to determine the reference range of BTM. INTRODUCTION: The aim of this study is to determine the reference intervals of BTMs for adult females in China with an attempt to reference the peak bone mineral density (BMD) with the corresponding BTM valley. METHODS: This study included 546 premenopausal and 394 postmenopausal women. The levels of several BTMs were determined, and the BMD was measured using a dual-energy X-ray absorptiometry. RESULTS: The BTMs of postmenopausal women were 17-96 % higher than premenopausal women. The change of BTM with age presented an optimal goodness-of-fit according to the cubic regression model (R (2) = 0.074-0.346, all P = 0.000). All kinds of BTM levels were positively correlated with age in premenopausal women aged 27-56 years old (r = 0.167-0.502, P = 0.023-0.000). Except for uCTX, the BTM reference value determined using a curve-fitting valley was significantly lower than the reference values for premenopausal women. The BTM reference values determined in this study were also significantly different from the reference values given by the manufacturers of the reagents used. CONCLUSIONS: This study found that the changes of level with age of BTMs in Chinese women present an optimal goodness-of-fit according to the cubic regression model. The fitting valley corresponds to the BMD fitting peak and may possibly be an effective means of determining the BTM reference intervals.

Effect of polymorphisms in the CSN3 (κ-casein) gene on milk production traits in Chinese Holstein Cattle.

This study was designed to evaluate significant associations between single nucleotide polymorphisms (SNPs) and milk composition and milk production traits in Chinese Holstein cows. Six SNPs were identified in the κ-casein gene using pooled DNA sequencing. The identified SNPs were genotyped by Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) methods from 507 individuals. Out of six, we identified three non-synonymous SNPs (g.10888T>C, g.10924C>A and g.10944A>G) that changed in the protein product. SIFT (Sorting_Intolerant_From_Tolerant) prediction score (0.01) demonstrated that protein changed Isoleucine > Threonine (g.10888T>C) will affect the phenotypes. Significant associations between identified SNPs and three yield traits (milk, protein and fat) and two composition traits (fat and protein percentages) were found whereas it did not reach significance for fat percentage in haplotypes association. Importantly, the significant SNPs in our results showed a large proportion of the phenotypic variation of milk protein yield and concentration. Our results suggest that CSN3 is an important candidate gene that influences milk production traits, and identified polymorphisms and haplotypes could be used as a genetic marker in programs of marker-assisted selection for the genetic improvement of milk production traits in dairy cattle.

Relationships between serum omentin-1, body fat mass and bone mineral density in healthy Chinese male adults in Changsha area.

PURPOSE: The present study is firstly designed to identify the relationship between serum omentin-1 concentration, body fat mass and bone mineral density in healthy Chinese male adults in Changsha city. METHODS: A total of 219 (20-80 years old) healthy subjects were enrolled in this cross-sectional study. Serum omentin-1, adiponectin, leptin, resistin and bone turn over biochemical markers were measured with enzyme-linked immunosorbent assay. Bone mineral density (BMD) and fat body composition were determined using dual-energy-X-ray absorptiometry. RESULTS: Serum omentin-1 levels in the overweight subjects were significantly lower than those of the subjects with normal weight (p < 0.05). Omentin-1 was negatively correlated with weight (r = -0.418), body mass index (BMI, r = -0.419), waist circumference (r = -0.402), waist-to-hip ratio (WHR, r = -0.355), fat body mass (FBM, r = -0.430), fat % (r = -0.408), trunk fat (-0.431). However, after controlling for age, BMI and FBM, no significant correlation was noticed between omentin-1 and BMD at different skeletal sites. Pearson's correlation coefficients and partial correlation coefficients after adjustment showed no significant correlations between omentin-1 and bone turn over biochemical markers, including bone-specific alkaline phosphatase and bone cross-linked N-terminal telopeptides of type I collagen. Multiple line stepwise regression analysis revealed that FBM, WHR, adiponectin were important variables affecting omentin-1. Moreover, lean tissue mass was the most important factor affecting BMD and explained 10.5-14.7 % of the variance. Omentin-1, leptin and resistin were not the predictors of BMD. CONCLUSIONS: Serum omentin-1 was negatively correlated with FBM and BMI in healthy Chinese male adults, It was not significantly correlated with bone turnover biochemical markers. Omentin-1 may exert ambiguous effects on BMD, which maybe caused by the complex interactions among adipokines, hormonal activity, and body composition and bone metabolism.

Effect of FASN gene on milk yield and milk composition in the Chinese Holstein dairy population.

Fatty acid synthase (FASN) is a multifunctional protein that catalyzes de novo synthesis of fatty acids in cells. It plays a key role in the lipid biosynthesis as well as in the general metabolism of all living animals. We herein investigated polymorphisms of FASN. As a result, six single nucleotide polymorphisms (SNPs) were found and then genotyped in 752 Chinese Holstein cows. It was found that g.17924A>G was non-synonymous, g.13965 C>T, g.16907 T>C and g.18663T>C were synonymous mutations and two other two SNPs, g.8948 C>T (ss491228481) and g.14439T>C (rs133498277), were in intronic sequences of the gene. All such identified SNPs were found to be associated with milk yield and composition traits (P = 0.0441 to <0.0001). Significant additive and allele substitution effects were observed for three yield traits at all six loci as well (P < 0.05 to <0.01). Complete linkage disequilibrium among the five SNPs, with the exception of g.8948 C>T, was observed.

Oral administration with attenuated Salmonella encoding a Trichinella cystatin-like protein elicited host immunity.

Trichinellosis is a public health problem and is regarded as an emergent/re-emergent disease in various countries. The cDNA encoding a cystatin-like protein (Ts-cystatin) was identified by immunoscreening intestinal muscle larvae cDNA libraries with serum from pigs experimentally infected with 20,000 Trichinella spiralis muscle larvae. To study its impact on host immunity, we chose a eukaryotic expression system based on several comparisons of immunogenicity between the two Salmonella typhimurium administration schemes, which indicated that the eukaryotic expression system was superior. Humoral IgG and mucosal IgA were measured to determine the antibody response. To explore whether Th1 and Th2 responses were responsible for the induced protection, Th1- and Th2-specific cellular transcription factors and the cytokine profile were examined. Changes in the T lymphocyte and macrophage populations were detected by flow cytometry. Lastly, parasitological examination was examined. The results showed that Ts-cystatin induced a Th1/Th2-mixed type of immune response and decreased STAT6 transcription. The intestinal adult recovery increased by 10.9% in the Ts-cystatin group, the Ts-cystatin group fecundity rate was decreased by 91%. Furthermore, the number of muscle larvae did not change compared with the control group. In conclusion, our results suggest that Ts-cystatin plays an important role in Trichinella resistance to rapid expulsion by the host and is worth further study.