Objective To investigate the relation between the cellular immune function and unexplained recurrent spontaneous abortion(URSA) and the mechanism of active immunotherapy on URSA patients.Methods The flow cytometry(FCM) was used to detect CD3+ ,CD4+ ,CD8+ T lymphocyte and CD16+CD56+ natural killer(NK) cell subsets and the ratio of CD4+/CD8+ of peripheral blood(PBL).112 cases with URSA(76 cases treated with active immunotherapy) and 30 cases of normal fertiled(NF) women were studied.The percentages of T lymphocyte and NK cell subsets before and after therapy were compared among 76 cases wtih URSA treated by active immunotherapy.The rate of next successful pregnancy of URSA patients treated with and without active immunotherapy was compared.Results The percentages of CD3+ and CD16+CD56+ cell subsets as well as the ratio of CD4+/CD8+ of the URSA patients were significantly higher than those of NF cases(P <0.05).After active immunotherapy,the percentages of CD3+ and CD56+CD16+ cell subsets as well as the ratio of CD4+/CD8+ of URSA cases were significantly decreased(P <0.05 ).The rate of next successful pregnancy of URSA cases with and without active immunotherapy were 88.2% and 31.2% respectively,the difference was significant(P< 0.05).Conclusion The changes in the percentages of T lympyocyte and NK cell subsets have something to do with URSA.Active immunotherapy can effectively regulate the cellular immune function and increase the rate of next successful pregnancy of URSA patients.

Seven compounds were isolated from Onychium japonicum by macroporous resin, silica gel, ODS, Sephadex LH-20 column chromatography and semi-preparative HPLC. Their structures were identified by NMR, MS and other spectroscopic methods as onychone A (1), quercetin (2), quercetin-3-O-α-L-rhamnoside (3), kaempferol-7-O-β-D-glucopyranoside (4), kaempferol-3-O-α-L-rhamnopyranoside (5), (-)-prunin (6), and norathyriol (7). Compound 1 is a novel macrocyclic flavonoid, and all the others are reported from this plant for the first time. In vitro cytotoxic activities of compounds 1-7 were evaluated by MTS testing with five cancer cell lines. Compound 7 exhibited weak cytotoxicity against tumor cell lines A549, SMMC-7721, and SW480.

We predicted the anti-hepatitis B virus (HBV) active components and mechanism of Salvia miltiorrhiza based on network pharmacology. The active components of S. miltiorrhiza were obtained through TCMSP, PubChem database and literature research. The potential targets of the active components and HBV infection were predicted by SwissTargetPrediction and GeneCards databases, respectively. The protein-protein interaction (PPI) network was constructed by String database. Cytoscape software was adopted to construct a visual network of active component-disease target and perform topological analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed using DAVID platform. The molecular docking of key components and core targets was carried out by AutoDock Vina software. We screened out a total of 38 active components and 178 disease-component overlapping targets. Enrichment analyses obtained 405 related GO items and 68 signaling pathways, such as T/B cell receptor signaling pathways, PI3K/AKT signaling pathway, and mTOR signaling pathway. According to the results of molecular docking, most characteristic components of S. miltiorrhiza (miltionone Ⅱ, miltirone, protocatechuic acid, lithospermic acid, protocatechualdehyde) showed good affinity with the key targets (PIK3CA, APP, STAT3,AKT1 and mTOR). Furthermore, the anti-HBV activity of lithospermic acid, the representative active component of S. miltiorrhiza, and its regulation on PI3K/AKT and mTOR signaling pathways were investigated in an HBV replicating mouse model. Animal welfare and experimental procedures follow the regulations of the Animal Ethics and Welfare Committee of Hubei University. The results showed that lithospermic acid significantly inhibited HBV DNA replication, reduced serum HBsAg and HBeAg levels, and decreased the phosphorylation protein expression levels of AKT and mTOR in liver, indicating that lithospermic acid might exert the anti-HBV activity by regulating PI3K/AKT and mTOR signaling pathways.

Objective To analyze the effect of dapagliflozin on epicardial fat attenuation(EFat)in patients with coronary heart disease and type 2 diabetes.Methods The patients with coronary heart disease and type 2 diabetes in our hospital were retrospectively collected.The patients were grouped into treatment group and control group.The treatment group was newly treated with dapagliflozin on the basis of the original treatment.The control group maintained the original antidiabetic drugs without using dapagliflozin.After median follow-up of thirteen months,the clinical characteristics,including body mass index(BMI),fasting blood glucose(FBG),glycosylated hemoglobin(HbA1c),triglycerides,total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),and EFat between the two groups were compared.Results A total of 235 patients were collected,of which 205 cases completed follow-up.There were 111 cases in the treatment group and 94 cases in the control group.The BMI,FBG and HbA1c of patients in the treatment group significantly decreased compared to those before therapy,while HDL-C significantly increased.The EFat of patients in the treatment group significantly decreased compared to baseline[(-96.48±14.71)HU vs(-88.59±10.60)HU,P＜0.05].Pearson correlation analysis showed a negative correlation between changes in EFat and changes in HDL-C in patients in the treatment group(r=-0.188,P＜0.05).Conclusion Dapagliflozin can reduce the epicardial fat attenuation in patients with coronary heart disease and type 2 diabetes.

Objective To evaluate the predictive value of Holter ECG recordings for patients with moderate-severe obstructive sleep apnea and hypopnea syndrome ( OSAHS ) . Methods Holter recordings was performed in 76 patients who were diagnosed OSAHS by polysomnography( PSG) within one month from Jan. 2008 to July 2009 in our hospital. Twenty-eight patients were identified as mild OSAHS (AHI≤20) and forty-eight patients were moderate-to-severe OSAHS ( AHI >20). The indexes of heart rate variability (HRV) , total scores of thirteen sleep apnea risk indexes of Holter recordings and BMI were analyzed by bivariate Logistic regression analysis. Results Clinical features ( eg. Gender, age, complicated with hypertension,coronary heart disease, diabetes mellitus, hyperglycemia, and taken β-blocker) , total scores, the sum of thirteen sleep apnea risk scores collected by Holter recordings (5. 64 ± 2. 33 vs. 6.42 ± 2. 22, respectively,P>0. 05 ) were similar between patients with mild OSAHS and moderate-to-severity OSAHS. VLF/Total Power>70% ,the difference of daytime/nighttime LF Power < -70 and BMI were independent predictors of moderate-to-severe OSAHS with OR 3. 98 (1. 087 - 14. 596), 3. 69 (1. 106 - 12. 285) and 1. 28 (1. 062 - 1. 544), respectively (all P < 0. 05). Conclusions VLF/Total Power and the difference of daytime/nighttime LF Power and BMI could be used as screening parameters to recognize patients with moderate-to-severe OSAHS.

Be subjected to study the application in the hypersaline organic wastewater treatment,A moder-ately halophilic bacterium YS-1 was isolated from the salty water of a Ludaokou salina of Weihai in China.The polyphasic taxonomy study was processed through identifying its morphological features by AFM,researched the features of culture and measured physiologic-biochemical index,analyzed 16S rDNA se-quence homology Phylogenetic analysis based on 16S rDNA gene sequences showed that the closest relative of the strain was Halomonas sp.(AB167061).Experimented of intensifying hypersaline organic wastewater treatmented processing in the SBR demonstrate a significant result.The wastewater salt content of initial wastewater was 12% and initial CODcr was 1494 mg/L.After 72 h,the removal efficiency of CODcr achieved 90.0% and after 120 h,it achieved 98.1%.The experiments also showed the hypersaline organic wastewater treatment and presented the feasibility of acclimating of halophilic bacteria and running of proc-essing system.

Objective The purpose of this study was to approach the relation of SNP43,SNP44 locus, main haplotypes and haplotype combinations with type 2 diabetes mellitus(T2DM).Methods According to the theory and principles of systematic review,data from case-control studies regarding the association between calpain-10(CAPN10) gene and T2DM were derived through electronic search of PubMed and Chinese journals databases.To gain a more precise estimation of the relationship,a stratified Meta-analysis with four subgroups was pertbrmed according to the races.Publication bias Was also assessed.Results The association with T2DM in different races was evaluated.In Mongoloid race,SNP43-G allele,G/G genotype and 111/221 haplotype combination showed notable association with T2DM with Ors (95%CI) as 1.368(1.155-1.620),1.437(1.186-1.741) and 2.762 (1.287-5.927) respectively.In Caucasoid race,SNP44-C allele,111/111 hapotype combination showed strong relationship with T2DM with Ors(95%CI) as 1.144(1.023-1.278),1.291(1.050-1.586) respectively.In Hybrid race,only one positive finding Was obtained which Was SNP44-C allele with OR(95%CI)as 1.653(1.025-2.665).Conclusion SNP43-G allele,G/G genotype,111/221 were risk factors to Mongoloid race.And SNP-C allele,111/111 haplotype combination were risk factors to Caucasoid race,and SNP44-C allele to Hybrid race.

Objective To explore the relationship between serum homocysteine and metabolic syndrome (MS).Methods A cohort with 1680 people involved in a community-based population in Beijing was investigated.Metabolic syndrome was defined by NCEP-ATP Ⅲ criteria.Multivariate logistic regression analysis was used to estimate the odds ratios (OR) of MS.Multiple linear regression analysis was performed to analyze the association between Hcy and characteristic variables.Results Homocysteine was higher in MS population compared to those without MS ( 17.99 μmol/L vs.17.18 μmol/L,P=0.007) after adjusted for age and sex.Levels of homocysteine increased with the presence of MS components (from 0 to 4 or 5) (16.71,16.94,17.62,18.20,17.82 μmol/L respectively,P=0.044 for linear trend).Among the components,groups with larger waist circumference,higher blood pressure and triglycerides showed significantly higher Hcy level than their counterparts.Results from multiple logistic regression analysis revealed that the highest Hcy quartile (Hcy Ⅳ ) was significantly associated with MS.Compared with the lowest Hcy quartile (Hcy Ⅰ ),the adjusted odds ratio of having MS in HcyⅣ was 1.379(1.005-1.892) after adjusting for age,sex,levels on creatinine/estimated glomerular filtration rate (eGFR)/low-density lipoprotein cholesterol (LDL-C) and uric acid,smoking,alcohol intake and exercise.In the partial correlation analyses,Hcy was positively associated with body mass index (BMI),waist circumsternece,blood pressure,LDL-C,triglycerides (TG),uric acid,serum creatinine,eGFR,but inversely associated with high-density lipoprotein cholesterol (HDL-C) and independently with age and sex.In multiple linear regression analysis,age,male sex,BMI,LDL-C,creatinine and uric acid were found to be independently associated with Hcy level.Conclusion There was an association noticed between the MS using NCEP-ATP Ⅲ criteria and the highest quartile level of Hcy in this study.Factors as age and being male,the levels of BMI,LDL-C,creatinine and uric acid were independently associated with the Hcy level.

To explore the gene-based principal component logistic regression model and its application in genome-wide association study.Using the simulated genome-wide single nucleotide polymorphism (SNPs) genotypes data,we proposed a practical statistical analysis strategy-'the principal component logistic regression model',based on the gene levels to assess the association between genetic variations and complex diseases.The simulation results showed that the P value of genes in related diseases was the smallest among the results from all the genes.The results of simulation indicated that not only it could reduce the degree of freedom through hypothesis testing but could also better understand the correlations between SNPs.The gene-based principal component logistic regression model seemed to have certain statistical power for testing the association between genetic genes and diseases in the genome-wide association studies.

Objective To investigate the effect of extracts from rabbit skins inflamed by Viccinia virus vaccine (analgecine) on the proliferation of human cancer cells and on the cytokine secretion in mouse spleen lymphocytes in vitro. Methods Five human tumor cell lines, HepG2, LM3, H460, A549 and HeLa were used and the effect of analgecine (1.63, 0.815, 0.326, 0.163 and0.0815 U/ml) on the cell proliferation was evaluated by the CCK-8 assay. The mouse spleen was isolated aseptically, and the spleen lymphocyte suspension was prepared and cultured with PRMI-1640 medium containing 10％ fetal bovine serum (FBS). For detection of the cytokine IL-2, IFN-γ, IL-4 and IL-12 level, the stimulant concanavalin A (ConA) or lipoplysaccharide (LPS) was added into the lymphocyte suspension, and the lymphocytes were cultured under the presence of analgecine at the final concentration of 0.815, 0.163 and 0.0815 U/ml for 24 hours. Then, the level of the cytokines in the supernatant was detected by the ELISA kit. On the other hand, the effect of supernatant of the spleen lymphocyte cultures under the presence of analgecine at 0.815 U/ml on the proliferation HepG2 cells was also evaluated by the CCK-8 assay. The CCK-8 assay was performed after cultivation of the HepG2 cells in the whole supernatant or in its dilution with fresh medium for 24 hours. Results Analgecine showed a dose-dependent inhibitory effect on the five tested cancer cell lines, with the inhibition rate of 58.95％, 55.08％, 57.28％, 45.80％ and 48.18％ at the 1.63 U/ml on the HepG2, LM3, H460, A549 and HeLa cells, respectively. Compared with the control group, the secretion of IL-2, IFN-γ and IL-4 was significantly increased in the 0.163 and 0.815 U/ml analgecine groups (P＜0.01), while the secretion level of IL-12 was increased in the 0.0815, 0.163 and 0.815 U/ml analgecine groups (P＜0.01). The supernatant of the mouse spleen lymphocyte cultures under the presence of0.163 U/ml analgecine could inhibit the HepG2 cell proliferation in a dose-dependent manner, and the inhibitory effect of the whole supernatant was significantly stronger than the effect of the same concentration analgecine 0.163 U/ml (P＜0.01). Conclusion Analgecine could inhibit the cell proliferation of the tested five human cancer cell lines, increased the secretion of IL-2, IFN-γ, IL-4 and IL-12 cytokines in mouse spleen lymphocytes, all in vitro, and its effect on the cytokine secretion may be related to the inhibitory effect on the human cancer cell proliferation.