ObjectiveTo evaluate the predictive value of the sperm quality to fertilization outcomes after short-time insemination.MethodsA total of 558 cycles of short-time insemination in the Reproductive Medical Center of Henan Provincial People's Hospital during January 2009 to June 2010 excluding patients aged ＞ 38 years and M Ⅱ oocyte number ＜ 3 were analyzed retrospectively.According to whether undergo rescue intracytoplasmic sperm injection( Re-ICSI),all cycles were divided into in vitro fertilization (IVF)group (472 cycles) and rescue intracytoplasmic sperm injection (Re-ICSI) group (86 cycles).Both IVFgroup and Re-ICSI group were subdivided into primary infertility and secondary infertility according to previous history of pregnancy.269 primary infertility cycles and 203 secondary infertility cycles were characterized in IVF group; and 64 primary infertility cycles and 22 secondary infertility cycles were characterized in Re-ICSI group.x2 test was applied for comparison of embryo plant rate,clinical pregnancy rate,early miscarriage rate between IVF and Re-ICSI groups,while Fisher test was used for comparison of live birth rate.and Mann-Whitney U test was utilized for comparison of duration of infertility,forward moving sperm counts,abnormal sperm rate,sperm acrosin activity between IVF and Re-ICSI groups.ResultsThe embryo plant rate,clinical pregnancy rate,early miscarriage rate,live birth rate of IVF group were:29.4％,44.9％,13.4％,37.0％ respectively; the above indicators in Re-ICSI group were:25.7％,34.6％,10.7％,29.6％ respectively,the differences of the indicators between the two groups had no statisticalsigmficance (x2 =0.869,2.963,0.010,P =0.351,0.085,0.922,0.098).Median of duration of infertility,forward moving sperm counts,abnormal sperm rate,sperm acrosin activity of primary infertility cycles in IVF group were:4.00(3.00 -6.00) years,58.37(33.64 - 102.27) × 106,81.09％ (79.41％ -88.69％ ),76.30 (48.50 - 92.46 ) μIU/106 sperm respectively ; in Re-ICSI group were:5.00 ( 3.25 -8.00) years,36.33 (20.59 -64.43 ) × 106,85.5％ (81.28％ - 89.02％ ),47.14( 31.61 -90.24) μIU/106 sperm respectively,the differences of them between the two groups had statistical significance (Z =-2.617, -3.505, -3.553, -3.530,P =0.009,0.000,0.000,0.000).Median of duration of infertility,forward moving sperm counts,abnormal sperm rate,sperm acrosin activity of secondary infertility cycles in IVF group were:5.00 (3.00 -7.00) years,63.00 (34.20 - 107.73 ) × 106,81.29％ (79.90 -86.09) ％,78.34 ( 53.87 - 98.00) μIU/106 sperm respectively,in Re-ICSI group were:5.00 ( 3.75 -7.00) years,28.80 ( 18.57 - 48.56 ) × 106,88.79％ ( 84.04 - 95.64 ) ％,54.70 ( 39.73 - 76.77 ) μIU/106 sperm respectively,the differences of them between the two groups showed statistical significance except duration of infertility (Z =- 0.338,- 3.505,- 3.553,- 3.530,P =0.735,0.000,0.000,0.006).ConclusionThe duration of infertility,forward moving sperm counts,abnormal sperm rate,sperm acrosin activity have predictive value of fertilization outcomes after short-time insemination.

Objective To explore the role of VEGF positive expression in tumor tissue in the prognosis of liver transplantation for hepatocellular carcinoma (HCC).Method Fifty cases of liver transplant recipients with HCC confirmed immunohistochemically were enrolled in this study.The MaxVisionTM two-step method was applied to detect the expression of vascular endothelial growth factor(VEGF),and the microvessel density (MVD) was measured in para-cancerous tissues by using DAB staining.The correlation of the VEGF tumor tissue in tumor tissue with Child-Pugh,MELD,tumor diameter and number,differentiation,MVD,Milan criteria and UCSF criteria for HCC liver transplantation was analyzed.Result In the HCC tissue,the VEGF positive expression rate was 52％(26/50).The one-year survival of recipients positive and negative for VEGF was 78％ and 100％,respectively,and one-year recurrence rate was 32％ and 12％,respectively,with the difference being significant (P =0.043 and P =0.048 respectively).The expression of VEGF was associated with Child-Pugh,tumor diameter,MVD,Milan criteria and UCSF criteria (P＜0.05 for all).Logistic regression analysis showed that low differentiation and VEGF positive expression were independent prognostic factors for HCC recurrence after liver transplantation.Conclusion VEGF has a certain reference value to judge HCC invasiveness and prognosis of liver transplantation.

Objective To analyze the differences of biochemical and immunological indicators in gender or Lee′s classification of IgA nephropathy ( IgAN) to provide laboratory evidence for clinical diagnosis and treatment of IgAN.Methods Retrospective cohort study.The information of biochemical and immunological indicators of 213 in-hospital patients which were admitted in Chinese PLA General Hospital in June to December, 2012.The data were collected and analyzed with t-test, non-parameter analysis, Pearson correlation analysis, ROC curves analysis and Logistic regression analysis according to gender or Lee′s classification ( Lee′s≤3 group and Lee′s>3 group).Results In this study, the average age and sex ratio of patients with IgAN was ( 35.0 ±10.6 ) years old and 2.04∶1.00.T-test and non-parameter analysis indicated homocysteine(HCY), immunoglobulin E (IgE), immunoglobulin M (IgM), prealbumin (PA), ceruloplasmin ( CP) ,α1-acidoglycoprotein(α1-AGP) , albumin ( Alb) ,α2-globulin(α2-G) andγ-globulin (γ-G) had significant difference in gender (P<0.05).And cystantin-C (CYS-C), HCY, complement 4 (C4),β2-microglobulin(β2-MG), PA, α1-AGP, α1-globulin(α1-G), immunoglobulin G (IgG), IgM and transferrin ( TF ) had significantly different in Lee′s classification ( P<0.05 ).ROC curves analysis showed that the areas under curves of CYS-C andβ2-MG were 0.891 and 0.839 respectively;the sensitivity was 90.2%and specificity was 75.3% when cutoff value of CYS-C was 1.56 mg/L; the sensitivity was 80.3%and specificity was 78.8% when cutoff value of β2-MG was 0.275 mg/dl.Logistic regression analysis showed that CYS-C ( OR: 31.380, 95% CI: 10.808 -91.113, P=0.000 ) and HCY ( OR:1.035, 95%CI:1.002-1.069, P=0.040) were introduced into the model after twice variable selections.ROC curve of CYS-C combined with HCY for classifying Lee′s classification showed that the sensitivity, specificity, Jorden index and AUC were 0.843, 0.862, 0.705 and 0.894 ( P=0.000 ) , respectively.Conclusions CYS-C,β2-MG, PA and HCY were valuable indicators for Lee′s classification.CYS-C andβ2-MG had high correlation with Lee′s classification, which indicated that these two tests were related to the severity of IgAN.CYS-C combined with HCY could improve the diagnostic efficiency of IgAN Lee′s classification.Combination of biochemical and immunological indicators could improve the accuracy of Lee′s classification and provide effective laboratory evidence for clinical treatment of IgAN.

This study was designed to investigate the correlation between autophagy and polarization of macrophages in atherosclerosis (AS) plaque in arteriosclerosis obliterans amputees. Femoral artery specimens from arteriosclerosis obliterans amputees were performed hematoxylin and eosin (HE) staining, oil red O and immunofluorescence staining to observe the morphology of atherosclerotic plaque, phenotype of macrophages and autophagy in plaque; using real-time quantitative RT-PCR technology to detect the mRNA level of M1 and M2 type markers in arterial tissue; to analyze polarized signal pathway and autophagy protein levels in macrophages by Western blotting. Arterial specimens staining showed obvious lipid deposition and obvious infiltration of amount of foam cells and inflammatory cells. Macrophages were mainly expression M1 type in percentage in fibrous plaque. Although both M1 and M2 macrophages were upregulated in atheromatous plaque, the increase was dominant in M2 type in percentage. The level of autophagy was significantly higher in the atheromatous plaque than that of fibrous plaque. The expression of tumor necrosis factor- α (TNF-α), monocyte chemotactic protein-1 (MCP-1), inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6) and interleukin-12 (IL-12) mRNA was significantly higher in fibrous plaque than that of atheromatous plaque (P < 0.01 or 0.05), and arginase-1 (Arg-1), transforming growth factor-β (TGF-β), CD163 and interleukin-10 (IL-10) mRNA was significantly lower than that in atheromatous plaque (P < 0.01). The levels of p-STAT1 and NF-κB were significantly increased in fibrous plaque (P < 0.01), while p-STAT6 expression was significantly increased in atheromatous plaque (P < 0.01). The level of LC3-II was significantly higher in atheromatous plaque than that in fibrous plaque (P < 0.01). Macrophages in early atherosclerotic plaque were induced to M1 type through p-STAT1/NF-κB pathway and expressed moderate levels of autophagy; while macrophages in advanced plaques were induced to polarization of M2 type through p-STAT6 pathway. M2 macrophages expressed a higher level of autophagy than M1 macrophages.
Amputees ; Arginase ; metabolism ; Arteriosclerosis Obliterans ; pathology ; Atherosclerosis ; pathology ; Autophagy ; Cell Polarity ; Chemokine CCL2 ; metabolism ; Foam Cells ; cytology ; Humans ; Interleukin-10 ; metabolism ; Interleukin-12 ; metabolism ; Interleukin-6 ; metabolism ; Macrophages ; cytology ; NF-kappa B ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Phenotype ; STAT6 Transcription Factor ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Up-Regulation

OBJECTIVETo investigate the role of Gemin3 in cell proliferation and its regulation pathway.

METHODSUsing co-immunoprecipitation and GST pull-down assay to determine the domain of Gemin3 and p53 binding and interaction in vitro and in vivo. To check the effect of Gemin3 on p53 by luciferase reporter assay. Stable Gemin3 knock-down cell lines were generated by lentivirus-delivered small hairpin RNA then puromycin selection. Real-time PCR was used to confirm the effect of Gemin3 level on p53 and its downstream genes, and flow cytometry was used to analyze the effect of Gemin3 on apoptosis.

RESULTSThe C-terminal of Gemin3 interacted with the DNA binding domain of p53. The p53 reporter gene, PA3M-p53 and increasing amount of GFP-Gemin3 were co-transfected into Saos-2 cells. Gemin3 repressed p53 expression at transcription level. Real-time PCR indicated that the expression of p53, p21 and Bax in Gemin3 knock-down cells was higher than that in the control cells. Western blot showed Gemin3 knock-down cells had a higher p53 espression. Flow cytometric assay showed that knock-down Gemin3 expression led to an increased cell apoptosis.

CONCLUSIONGemin3 binds with p53 forming a complex and plays an anti-apoptotic role by repressing the p53 expression.

Apoptosis ; B-Lymphocytes ; cytology ; Cell Line, Tumor ; DEAD Box Protein 20 ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Gene Knockdown Techniques ; Genes, Reporter ; Humans ; Immunoprecipitation ; Lentivirus ; genetics ; Osteosarcoma ; genetics ; metabolism ; pathology ; Plasmids ; Protein Binding ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transfection ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

The aim of this study was to investigate the effects of tyrosine kinase inhibitor PTK787 on cell proliferation, cell cycle and the expression of fak mRNA of human chronic myeloid leukemia (CML) cell line K562, and to explore the mechanism of PTK787 against acute myeloid leukemia. The MTT method was used to detect the effects of PTK787 in various concentrations and at different time points on proliferation of K562 cells; the flow cytometry was used to determine the effects of PTK787 in different concentrations on cell cycle of K562 cells; the RT-PCR was used to assay the expression of fak mRNA in K562 cells treated with PTK787 for 48 hours. The results showed that along with increasing of the concentration and prolonging of time, the inhibitory rate of PTK787 on K562 proliferation was gradually enhanced. The comparison between various concentration groups at same time or comparison between various time groups in same concentration showed significant differences (p < 0.05), in which the effect of 320 micromol/L PTK787 on cells was strongest, while the continuous increase of PTK787 concentration or prolong of action time did not enhance the inhibitory rate on K562 proliferation. With increasing of drug concentration, the cell proportion in G(1) phase gradually increased, the cell proportion in S phase gradually decreased, the comparison between various groups revealed significant differences (p < 0.05), however the continuous increase of drug concentration from 160 micromol/L did not obviously change the cell proportion in phases of cell cycle. With increasing of drug concentration, the expression of fak mRNA in K562 cells gradually reduced with significant differences between various groups (p < 0.05), but with continuous increase of drug concentration from 160 micromol/L, the effect of PTK787 on the expression of fak mRNA in K562 cells also did not obviously change. It is concluded that the PTK787 shows effect of anti-leukemia cells through inhibiting transformation of the K562 cells from G(1) phase into S phase and decreasing the expression of fak mRNA in cells.
Cell Cycle ; Cell Proliferation ; drug effects ; Focal Adhesion Kinase 1 ; genetics ; Gene Expression Regulation, Leukemic ; Humans ; K562 Cells ; Phthalazines ; pharmacology ; Pyridines ; pharmacology ; RNA, Messenger ; genetics

OBJECTIVETo study the metabolic difference of body influenced by active stress and passive stress under special events.

METHODSTo detect serum multiple biochemistry index of 57 earthquake rescue medical team and 13 victims of a natural calamity in Wenchuan earthquake by using Hitachi 7600 automatic analyzer.

RESULTSStress affected biochemistry index deeply. To compared with rescue medical team, the serum ADA, ALP and TG of victims increased obviously and TP, ALB, MAO, Cr, UA, K, Na, Cl, Ca, ApoA1 and HDL decreased obviously.

CONCLUSIONMany biochemistry index have been changed under stress and it relate with stress extent. The human body function status was better in active stress than in passive stress.

Blood Chemical Analysis ; China ; Disasters ; Earthquakes ; Humans ; Metabolism ; physiology ; Rescue Work ; Stress, Physiological ; physiology

Objective:To investigate the efficacy of Dahuang Zhechong pill combined with tenofovir disoproxil fumarate in treating liver fibrosis of chronic hepatitis B and its effect of intestinal flora.Methods:One hundred and twenty chronic hepatitis B cases with liver fibrosis treating in Rugao Hospital Affiliated to Nantong University from March 2018 to November 2019 were randomly divided into the control group and the observation group with 60 cases in each group according to the random number table. The control group was orally given with tenofovir disoproxil fumarate (1 tablet/time and quaque die).The observation group was treated with Dahuang Zhechong pill(3 g/time and bis in die) at basis of control group. The course was for 48 weeks for two groups. Liver function, liver fibrosis, hepatitis B virus e antigen(HBeAg) negative conversion rate, hepatitis b virus (HBV)-DNA negative conversion rate, the efficacy and intestinal flora changes were compared between the two groups before and after treatment. The adverse reactions were recorded in two groups.Results:After treatment, the levels of aspartic transaminase(AST), alanine aminotransferase (ALT), gamma-glutamyltransferase(GGT), total bilirubin (TBIL) in observation group were lower than those in control group [(41.62 ± 5.93) U/L vs. (50.41 ± 7.12) U/L, (41.99 ± 5.76) U/L vs. (52.17 ± 7.09) U/L, (34.46 ± 5.31) U/L vs. (49.41 ± 6.23) U/L, (20.35 ± 3.04) μmol/L vs. (25.60 ± 3.95) μmol/L], and the differences were statistically significant ( P<0.05). After treatment, the levels of hyaluronic acid (HA), procollagen Ⅲ (PCⅢ), type Ⅳ collagen (Ⅳ-C), Laminin(LN) in observation group were lower than those in control group [(167.45 ± 18.75) μg/L vs. (209.44 ± 22.96) μg/L, (103.44 ± 12.75) μg/L vs. (140.08 ± 16.33) μg/L, (81.41 ± 10.06) μg/L vs. (126.36 ± 14.94) μg/L, (108.41 ± 12.72) μg/L vs. (169.41 ± 19.22) μg/L], and the differences were statistically significant ( P<0.05). The total effective rate in the observation group was higher than that in the control group [91.67%(55/60) vs. 76.27%(45/59)], and the difference was statistically significant ( P<0.05). The negative conversion ratio of HBeAg, HBV-DNA in the observation group were higher than those in the control group [28.33%(17/60) vs. 11.86%(7/59), 83.33%(50/60) vs. 66.10%(39/59)], and the differences were statistically significant ( P<0.05). Compared with the control group, the levels of bifidobacteria and lactobacillus in the observation group were higher and the levels of escherichia coli and enterococcus in observation group were lower, and the improvement of intestinal flora in the observation group were better than those in the control group ( P<0.01). Conclusions:Dahuang Zhechong pill combined with tenofovir disoproxil fumarate in treating liver fibrosis of chronic hepatitis B can improve the liver function and liver fibrosis, and can improve the serological conversion rate of HBeAg, inhibit HBV DNA replication, increase the therapeutic effect, and is safety. The effect of Dahuang Zhechong pill on intestinal flora may be related to the curative effect.

Objective: To assess the feasibility of coronary lfow reserve (CFR) detection by SPECT myocardial perfusion imaging using a self developed software with preliminary clinical veriifcation. Methods: CFR calculation software was developed according to Mat lab guide. A total of 16 patients were enrolled including 13 male and 3 female at the mean age of (58±11) years . CAG conifrmed that 25 coronary branches were with stenosis>50% and 23 branches were without stenosis. 2-day ATP/rest99mTc-sestamibi dynamic SPECT myocardial perfusion imaging was conducted to detect CFR. First transit counts were used to sketch the interested pulmonary artery segments and to obtain the arterial input curve of contrast agent as total PAC reached to heart. Reconstructed short-axis images were divided into 3 sections to sketch interested territories (ROI) and to obtain RMC at each territory. Estimated CFR was expressed by the ratio of MBF=RMC/PAC followed by calculating the ratio of MFR=MBFstress/MBFrest. Results: The difference between simulated value and true value could be ignored which conifrmed that our program may accurately measure CFR. The reproducibility by different operators (r=0.986) and the same operator (r=0.983) was good. CFR value in non-stenosis branches were higher than stenosis branches (1.28 ± 0.19) vs (1.10 ± 0.27),P=0.008 and CFR value in stenosis branches was negatively related to stenosis degree (r=-0.5,P=0.02). Conclusion: Our self developed software is reliable for CFR detection by SPECT myocardial perfusion imaging; preliminary study showed good application prospect in clinical practice.

Objective To observe the clinical efficacy and safety of icotinib tablets in the treatment of stage Ⅲ A-N2 non-small cell lung cancer (NSCLC).Methods A total of 56 patients with stage ⅢA-N2 NSCLC were randomly divided into control group and treatment group with 28 cases per group.The control group was treated with 60-75 mg · m-2 docetaxel,once a week,intravenous drip,or 500 mg · m pemetrexed,once every 3 weeks,intravenous drip,discontinuation of treatment when the disease progresses or adverse drug reactions cannot be tolerated.The treatment group was treated with icotinib tablets 125 mg,tid,orally,discontinuation of treatment when the disease progresses or adverse drug reactions cannot be tolerated.The clinical efficacy,peripheral blood dendritic cell subsets,and adverse drug reactions were compared between two groups.Results After treatment,the objective remission rates of the treatment group and the control group were 89.29％ (25 cases/28 cases) and 71.43％ (22 cases/28 cases),the difference was statistically significant (P ＜ 0.05).After treatment,the number of dendritic cells (DC) in the treatment group and the control group were (15.44 ± 2.08) and (12.83 ± 1.71) × 106/L,the ratio of dendritic cells to mononuclear cells (DC/PBMC) were (0.62 ± 0.08) ％ and (0.57 ± 0.07) ％,the differences were statistically significant (P ＜ 0.05).The adverse drug reactions of two groups were rashes,diarrhea and mild liver dysfunction.The total incidences of adverse drug reactions in the treatment group and the control group were 46.43％ and 42.86％ without significant difference (P ＞ 0.05).Conclusion The clinical efficacy of icotinib tablets in the treatment of stage ⅢA-N2 NSCLC was exact,and it did not increase the incidence of adverse drug reactions.