Objective To study the effects of pulse high volume hemofiltration (PHVHF) on the changes of Th17 cells (T helper 17 cells) and CD4 + CD25 + reguratory T cells (Treg cells) in peripheral blood of patients with sepsis and to evaluate the clinical value of this intervention. Methods The patients were included in this prospective study as per the criteria of sepsis set by America Chest Physicians College/America Society for Critic Care Medicine in 1992. The patients were excluded: ① immune system disorder, ② acute stroke, ③ myocardial infarction, ④ virus hepatitis,⑤ human immunodeficiency virus infection, ⑥ under immunosuppressive therapy. Forty patients (24 males, 16 females, aged from 25 to 75years) with sepsis in ICU were enrolled from January. 2008 to November. 2010. According to the severity of disease, the patients were divided into three groups; moderate sepsis group (n = 14, 8 males, 6 females) , severe sepsis group (n = 15, 9 males, 6 females) , and septic shock group (n = 11, 7 males, 4 females). The initially clinical data of three groups were comparable. Twenty healthy individuals served as controls. According to the mode of treatment, forty patients were also divided into two groups: conventional treatment group (group A, n= 15) in which patients were treated without PHVHF within 5 days after admission and trial group (group B, n=25) in which patients were treated with pulsed high volume hemofiltration (PHVHF) within 5 days after admission. In group B, high volume hemofiltration (70 mL · kg-1 · h-1) was given to patients for 6 ～ 8 hours, and then conventional continuous vein - vein hemofiltration (35 mL · kg-1 · h-1) for 16 ~ 18 hours. The total length of period for continuum blood scavenging was 24 hours as one cycle. The interval between two cycles of blood scavenging was 24 hours. The changes of Th17 cells and CD4+ CD25 + Treg cells of 40 patients were detected with flow cytometry on the 1st day and the 5th day after admission. The data were analyzed by using SPSS version 13. 0 software. Measurement data were analyzed with Paired-samples t-test, independent-samples t-test or one way ANOVA . Ratio of small samples was compared with fisher's exact test, and the correlation was analyzed by using Pearson correlation analysis. Results The rates of Th17 cells were( 0.91 ±0.38)%, (2.09 ±0. 53)% , (3.90 ±0. 80)% , and ( 1. 85 ±0.35)% in control, moderate sepsis, severe sepsis, and septic shock groups, respectively, while the rates of CD4+ CD25+ Treg cells were (0.39 ±0.23)%, (1. 72 ±0. 59)% , (2.72 ±0. 22)% , and (3. 55 ±0. 51)% , respectively. The rate of Thl7 cells on the 1st day was higher in severe sepsis group than that in other two groups ( P < 0. 05 ) without significant difference between septic shock and moderate sepsis groups ( P > 0. 05). Moreover , the rate of CD4+ CD25 + Treg cells was up - regulated on the 1st day in the following order from high to low: septic shock group > severe sepsis group > sepsis group (P < 0.05). The rates of Th17 cells and CD4 + CD25 + Treg cells in patients of group B decreased in greater degree than that did in patients of group A (P < 0.05 ). Conclusions The changes of Th17 cells and CD4 + CD25 + Treg cells may play an important role in pathogenesis of sepsis, and the pulsed high volume hemofiltration may be one of the effective treatments for the patients with sepsis by regulating the rates of Thl7 cells and CD4 + CD25 + Treg cells.

Objective To identify a aldehyde dehydrogenases positive (ALDH+) cancer stem cell subpopulation in MCF-7 cells and to investigate the proliferation and differentiation characteristics of these cells in vitro.Methods ALDH+ breast cancer stem cells were isolated from MCF-7 cells by flow cytometry and the biological property of ALDH+ breast cancer stem cells were examined by scarification test,MTT,growth curvature and Transwell migration assay.Results The ratio of CD-/low24 CD+44 cells was about 1.4 ％ in MCF-7 cells.The ratio of ALDH+ CD-/low24CD+44 cells was about 1.2 ％.The growth curvature of ALDH+ breast cancer stem cells was almost the same with that of CD-/low24 CD+44 cells.The distance between cells was obviously shorter in both CD-/low24 CD+44 cells scarification zone and ALDH+ CD-/low24 CD+44 cells scarification zone.The migration ability of CD-/low24CD+44 cells and ALDH+ CD-/low24CD+44 cells was stronger than control group cells.There were migration ability differences between CD-/low24CD+44 cells and ALDH+ CD-/low24CD+44 cells.The results of Transwell experiments were in coincidence with above results.Lots of CD-/low24CD+44 cells and ALDH+ CD-/low24CD+44 cells were through the membrane.In MTT assay,absorbance values were 1.05±0.098,1.56±0.075 and 1.67±0.032.Conclusion CD--/low24CD+44 and ALDH+ CD-/low24CD+44 breast cancer stem cell subpopulation exist in MCF-7 cells.ALDH could potentially be used as a molecular marker to identify breast cancer stem cell subpopulation.

ObjectiveTo investigate the correlation between serum CA125 level and the severity of liver dysfunction in patients with liver cirrhosis. MethodsWanfang Data, CNKI, CBM, and VIP were searched for Chinese articles on the correlation between serum CA125 level and the severity of liver dysfunction in patients with liver cirrhosis published from January, 2008 to October, 2018, with a liver cirrhosis group and a normal control group in each article. Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) was used for quality assessment. The mean and standard deviation of CA125 in liver cirrhosis group, healthy control group, and liver cirrhosis groups with different Child-Pugh classes were analyzed. Meta-Analyst software was used to calculate the standardized mean deviation (SMD) of CA125 in each group and perform the meta-analysis. A heterogeneity analysis was performed for the studies included in this study; a random effects model was used in case of significant heterogeneity, while a fixed effect model was used in case of insignificant heterogeneity. A one-way analysis of variance was used for comparison of continuous data between multiple groups. ResultsA total of 15 articles were included in this study. The meta-analysis showed that the liver cirrhosis group had a significantly higher serum CA125 level than the healthy control group (181.18±110.76 U/ml vs 15.10±7.15 U/ml, SMD=2.28, 95% confidence interval: 1.81-2.76, P＜0.001). The level of CA125 increased significantly with the increase in Child-Pugh class (F=15.704, P＜0.001). ConclusionSerum CA125 level is correlated with the severity of liver dysfunction in patients with liver cirrhosis and thus has a certain value in evaluating the severity of liver dysfunction and predicting prognosis.

Objective To explore the effect of “net type” management in the prevention and control of multi-drug resistant organisms ( MDROs) .Methods Totals of 38 MDROs infected patients were selected from the department of geriatrics psychiatry , neurology and rehabilitation medicine from 2012 to 2013 .Totals of 71 cases from January to December 2012 were selected as the control group , and were given the traditional prevention and control management .Totals of 67 cases from January to December 2013 were selected as the experiment group , and were given the “net type” management . Nosocomial infection , underreporting , implementing prevention and control measures were compared between the two groups .Results The percentages of underreporting , input outside the hospital , nosocomial infection of the experiment group after using the “net type” management were 5.97%, 39.44% and 43.28%, respectively, those percentages of the control group were 35.21%, 39.44%and 61.97%, and the differences were statistically significant (χ2 =16.040,9.548, 4.108,respectively;P <0.05).The medical orders for separation, setting separation signs, correct use of medical supplies , specific application , material dispose , correct dispose of medical waste and the incidence of health education were 89.55%, 92.54%, 76.12%, 88.06%, 88.06%, 92.54% and 92.54% in the experiment group, which were significantly better than those of the control group (χ2 =16.178, 17.646, 39.613, 36.178, 17.006, 30.379, 40.519, respectively;P <0.05 ).Conclusions The “net type”management in secondary hospitals is better than the traditional method in the prevention and control of multi -drug resistant bacteria .

OBJECTIVETo study the mechanism underlying the effect of the SOCS3 rs4969170 A/G alleles on the occurrence of insulin resistance (IR) in patients with chronic hepatitis C.

METHODSThe promoter region of the SOCS3 gene was amplified by PCR,and luciferase expression vectors were constructed and transfected into HepG2,Huh7 cell lines.The relative luciferase activity of each expression vector was assessed by the dual luciferase reporter gene assay system.Western blotting was used to detect SOCS3 protein expression in PBMCs from groups of patients with the rs4969170 AA and AG genotypes.The state of IR in eight patients was evaluated by determining their HOMA-IR.

RESULTSThe pGL3-A, PGL3-G and pGL3-control vectors showed significantly different luciferase expression in the HepG2 cells (0.121 00 ± 0.022 07,0.027 00+/-0.012 49 and 0.043 33 ± 0.005 51; F =48.068, P=0.001) and in the Huh7 cell lines (0.164 70 ± 0.007 10,0.027 33 ± 0.017 04 and 0.033 67 ± 0.014 98; F =115.137, P=0.001). The expression of SOCS3 protein was significantly higher in the rs4969170 AA genotype group than in the AG genotype group (1.22 ± 0.40 vs. 0.30 ± 0.19; t =4.149, P=0.006).The IR index of patients with the rs4969170 AA genotype and the AG genotype was 4.11 ± 2.62 and 1.47 ± 1.01 respectively.There were three patients with IR in the rs4969170 AA genotype group and one in the rs4969170 AG group. There was no statistically significant difference between the two genotype groups (t=1.881, P=0.109).

CONCLUSIONSThe SOCS3 rs4969170 A haplotype may enhance transcriptional activity of the gene promoter to regulate gene expression, thereby increasing intracellular SOCS3 protein level and ultimately interfering with insulin signaling and causing IR in patients with chronic hepatitis C.

Cell Line, Tumor ; Genes, Reporter ; Genotype ; Haplotypes ; Hepatitis C, Chronic ; Humans ; Insulin Resistance ; Luciferases ; Polymorphism, Single Nucleotide ; Suppressor of Cytokine Signaling 3 Protein ; Suppressor of Cytokine Signaling Proteins

BACKGROUND/AIMS: Several studies have demonstrated that serum interferon-γ-inducible-protein-10 (IP-10) levels at baseline and single nucleotide polymorphisms (SNPs) near the IL28B gene were associated with viral response and treatment outcomes. Our purpose was to assess the combination of pretreatment IP-10 levels with IL28B SNPs as predictors of treatment response to pegylated interferon α-2a plus ribavirin in patients infected with genotype 1 hepatitis C virus in China. METHODS: Seventy-two patients with chronic hepatitis C without fibrosis/cirrhosis were enrolled in the study. The virologic parameters and baseline serum IP-10 levels were determined. IL-28B genotypes were determined by sequencing. RESULTS: In this cohort, serum baseline IP-10 levels lower than 426.7 pg/mL could predict rapid virological response/sustained virological response (SVR). Patients carrying favorable IL28B SNP genotypes had higher SVRs than did those carrying unfavorable variants (IL28B rs12979860, p=0.002; IL28B rs8099917, p=0.020). Combining both baseline IP-10 and IL28B SNPs could improve the prediction of SVR in favorable allele carriers of IL28B, rs12979860 CC and rs8099917 TT. Serum baseline IP-10 levels and IL28B genotypes were independent predictors of SVR. CONCLUSIONS: Our study shows that the combination of baseline serum IP-10 levels and the determination of IL28B SNPs increase the predictability of SVR rates in this cohort.
Alleles ; China* ; Cohort Studies ; Genotype* ; Hepacivirus* ; Hepatitis C* ; Hepatitis C, Chronic ; Hepatitis* ; Humans ; Interferons ; Polymorphism, Single Nucleotide ; Ribavirin

Objective To investigate the association of serum uric acid (sUA)-to-creatinine (Cr) ratio with nonalcoholic fatty liver disease (NAFLD). Methods A retrospective analysis was performed for the clinical data of 97 patients with NAFLD (NAFLD group) who attended Beijing Tiantan Hospital, Capital Medical University, from January to December 2020, and according to the results of abdominal ultrasound, they were divided into mild group with 33 patients, moderate group with 31 patients, and severe group with 33 patients. Related data were also collected from 36 healthy adults (control group) who underwent physical examination in our hospital during the same period of time. The above groups were compared in terms of sex, age, fasting blood glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL), gamma-glutamyl transpeptidase (GGT), sUA, serum Cr, and sUA/Cr ratio. The independent samples t - test was used for comparison of normally distributed continuous data between two groups, and an analysis of variance was used for comparison between three groups; the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups; the chi-square test was used for comparison of categorical data between groups. The Spearman test was used for correlation analysis, and a multivariate logistic regression analysis was used to investigate the risk factors for NAFLD. Results Compared with the control group, the NAFLD group had significantly higher levels of ALT, AST, TG, GGT, sUA, and sUA/Cr ratio ( Z =-4.881, -4.616, -4.221, and -3.563, t =12.974 and 10.710, all P < 0.05) and a significantly lower level of HDL ( Z =-5.682, P < 0.05). The severity of NAFLD (mild, moderate or severe) was positively correlated with ALT, TC, and LDL ( r =0.291, 0.272, and 0.253, all P < 0.05). The multivariate logistic regression analysis showed that sUA/Cr ratio was an independent risk factor for NAFLD (odds ratio=1.885, 95% confidence interval: 1.162-3.06, P < 0.05). Conclusion There is a significant correlation between sUA/Cr ratio and NAFLD, and sUA/Cr ratio is an independent predictive factor for NAFLD. The sUA/Cr ratio can be monitored to predict the onset of NAFLD, so as to achieve early identification, early diagnosis, and early treatment and improve prognosis.

Objective:To investigate the effect of ethephon exposure on sperm quality of adolescent male SD rats and the influence mechanism.Methods:A total of 40 45-day-old male SD rats were divided into control group and low, middle and high experimental groups according to the random number table method, 10 rats in each group.The said 4 groups were given 9 g/L normal saline, 100 mg/kg, 200 mg/kg, and 400 mg/kg ethephon aqueous solution for 28 days, respectively.One epididymal tail was taken to prepare sperm suspension, the sperm concentration and motility were detected.The testis and epididymis tissues were stained with HE, and their pathological changes were observed under light microscope.The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and the malondialdehyde (MDA) content in the testis were detected.Enzyme linked immunosorbent assay kit was used to mea-sure the epididymal α-glucosidase activity, L-carnitine (LC) content, nuclear factor erythroid 2-related factor 2 (Nrf2) and organic cation transporter 2 (OCTN2) expression levels.Then the oxidative damage caused by ethephon to epididymis was evaluated.SPSS 26.0 software was used for data analysis.Data were compared by One- way ANOVA among groups and LSD method between 2 groups. Results:The sperm concentration of the control group, low, medium and high dose groups were (40.21±1.94)×10 9/L, (35.23±2.53)×10 9/L, (23.61±2.62)×10 9 /L, and (18.86±2.16)×10 9 /L, respectively.The sperm activity rate were (70.98±3.01)%, (57.96±3.75)%, (45.71±2.41)%, and (31.23±2.26)%, respectively.The concentration and vitality of epididymal sperms in the experimental group were significantly lower than those in the control group (all P<0.01). In the control group, low, medium and high dose groups, the SOD activity were (46.48±2.21) U/mg prot, (38.49±2.56)U/mg prot, (33.80±1.73) U/mg prot, and (27.65±2.05) U/mg prot, respectively.The GSH-Px activity in said 4 groups were (21.41±1.95) U/mg prot, (17.32±1.28) U/mg prot, (15.09±0.94) U/mg prot, and (14.08±1.23) U/mg prot, respectively.The MDA content in said 4 groups were (1.41±0.09) nmol/mg prot, (1.59±0.09) nmol/mg prot, (1.81±0.09) nmol/mg prot, and (2.16±0.14) nmol/mg prot, respectively.Compared to the control group, the experimental groups had significantly lower SOD and GSH-Px activities and significantly higher MDA content (all P<0.05). α-glucosidase levels in the control group, low, middle and high experimental groups were (15.46±0.71) U/mL prot, (12.95±0.72) U/mL prot, (11.34±0.65) U/mL prot, and (8.76±0.60) U/mL prot, respectively.LC levels in the control group, low, middle and high dose groups were(6.21±0.31) μg/L, (5.89±0.13) μg/L, (5.02±0.12) μg/L, (4.38±0.07) μg/L, respectively, compared with those of the control group, the concentration of α-glucosidase and LC in experimental groups decreased significantly (all P<0.01). The expression levels of Nrf2 in epididymis of the control group, low, middle and high dose groups were (1.34±0.05) ng/L, (1.25±0.04) ng/L, (1.08±0.06) ng/L, (0.92±0.04) ng/L, respectively; the expression levels of OCTN2 in epididymis of the control group, low, middle and high dose groups were (4.55±0.12) ng/L, (4.23±0.11) ng/L, (3.20±0.24) ng/L, (2.59±0.05) ng/L, respectively, compared with those of the control group, the expression levels of Nrf2 and OCTN2 in experimental groups decreased significantly (all P<0.01). Conclusions:Ethephon exposure leads to excessive generation of reactive oxygen and oxidative stress in reproductive organs.Ethephon exposure may activate the Keap1-Nrf2/ARE signal pathway, resulting in a decrease in the number, vitality and quality of sperms, and impaired fertility.

Objective To investigate the effect and related mechanism of CD137 stimulation on aortic atherosclerotic plaque calcification in high fat diet fed ApoE-/-mice and on calcification of vascular smooth muscle cells (VSMCs).Methods (1) ApoE-/-mice fed with high fat diet were randomly divided into 3 groups:CD137 activated group (treated by 200 μg CD137 agonist i.p.once per week for 6 weeks,n =5);CD137 inhibited group (anti-CD137 group:200 μg anti-CD137 antibody + 200 μg CD137 agonist,i.p.,once per week for 6 weeks,n =5) and control group (n =5).Von kossa staining was used to observe the calcification of the aortic plaque and VSMCs.Immunohistochemistry was used to observe the expression of BMP-2 and Runx2 which are known mediators of osteogenic differentiation.(2) The mouse aortic VSMCs were obtained by Patch-attaching method.The calcium content was measured by Methylthymol Blue complexone method.The mRNA expressions of bone morphogenetic protein 2 (BMP-2) and Runx2 were measured by real-time fluorescent quantitative PCR (RT-PCR).The protein levels of BMP-2,Runx2 of the VSMCs were determined by Western blot.Results (1) In vivo,the plaque calcified area in ApoE-/-mice was significantly larger in CD137-agonist group than that in control group ((1.75 ± 0.33) × 104 μm2 vs.(0.23 ±0.07) × 104 μm2,P ＜0.01),and this effect was significantly reduced by cotreatment with CD137-antagonist ((0.83 ± 0.30) × 104 μm2 vs.(1.75 ± 0.33) × 104 μm2,P ＜ 0.05).The levels of BMP-2 and Runx2 were all significantly upregulated in CD137-agonist group than in control group (both P ＜ 0.01),again,this effect was blocked by cotreatment with CD137-antagonist (P ＜0.05).(2) Consistent with the in vivo results,VSMCs calcification was also more serious in CD137-agonist group than in control group,which could be significantly attenuated by eotreatment with CD137-antagonist.In VSMCs,calcium content level in CD137-agonist group was higher than in control group ((0.001 3 ± 0.000 2) mmol/mg protein vs.(0.000 7 ±0.000 1) mmol/mg protein,P ＜ 0.01),which could be significantly reduced by co-treatment with CD137-antagonist ((0.000 9 ± 0.000 2) mmol/mg protein vs.(0.001 3 ± 0.000 2) mmoL/mg protein,P ＜0.01).The mRNA and protein levels of BMP-2 and Runx2 were significantly upregulated in CD137-agonist group compared with the control group (P ＜ 0.05),which could be significantly down-regulated by cotreatment with CD-137 antagonist (P ＜ 0.05).Conclusion CD137 activation can promote vascular calcification in high fat diet fed ApoE-/-mice both in vivo and in vitro.

Objective To explore whether CD137-CD137L signaling can promote angiogenesis in atherosclerosis plaque via activating nuclear factor of activated T cells c1 (NFATc1).Methods Apolipoprotein E knock out mice were divided into the following groups:control group (n =5),CD137 activated group(n =5) and CD137 inhibited group (n =5).Immunohistochemistry was performed to detect the expression of CD31 in aortic plaque.Endothelial cells (bEnd.3) were purchased from ATCC and divided into the following groups:control group,IgG isotype control group,CD137 activated group and CD137 inhibited group.Western blot was used to determine total protein and nucleoprotein expression of NFATc1.The expression level of CD137 protein on the surface of endothelial cells was detected by flow cytometry (FCM) and CD137 protein of lysate of endothelial cells was detected by enzyme-linked immunosorbent assay (ELISA).Transwell assay was used to observe the migration ability of endothelial cells.Matrigel tube formation ability of endothelial cells were tested in the following groups:control group,CD137 activated group,silent NFATc1 + CD137 activated group,CD137 inhibited group,and over expressed NFATc1 + CD137 inhibited group.Results (1) In vivo,the expression level of CD31 was significantly higher in the aortic plaque of CD137 activated group than in control group(1 191 ± 187 vs.115 ± 30,P ＜ 0.05),while which was significantly downregulated in CD137 inhibited group (450 ± 92,P ＜ 0.05).(2) The level of nucleoprotein(3.07 ±0.03 vs.1.00 ±0.00,P ＜0.05) and total protein(2.18 ± 0.30 vs.1.00 ±0.00,P ＜0.05) of NFATc1 were significantly higher in CD137 activated group than in IgG isotype control group.The level of nucleoprotein(0.82 ±0.04) and total protein(0.84 ± 0.09) of NFATc1 were significantly lower in CD137 inhibited group than in CD137 activated group(both P ＜ 0.05).(3) FCM results showed that the fluorescence intensity of CD137 on the cell membrane was significantly higher in endothelial cells stimulated by TNF-α than in normal endothelial cells(5 163 ± 329 vs.1 660 ± 162,P ＜ 0.05).(4) ELISA examination showed that the level of CD137 protein was significantly higher in endothelial cells stimulated by TNF-α than in normal endothelial cells ((573.4 ± 23.7) pg/mg vs.(69.5 ± 16.7) pg/mg,P ＜ 0.05).(5) Migration cell number was remarkably higher in CD137 activated group than in IgG isotype control group(1.19 ±0.13 vs.1.00 ±0.00,P ＜0.05) and significantly lower in CD137 inhibited group(0.82 ± 0.06) than in control group (P ＜ 0.05).(6) Values of the formation of the tube length ((5.76 ± 0.18) mm vs.(4.21 ± 0.11) mm,P ＜ 0.05) and branch number (29.38 ± 1.28 vs.21.13 ± 0.96,P ＜0.05) were both significantly higher in CD137 activated group than in the control group.The formation of the tube length ((1.90 ±0.11)mm) and branch number(8.91 ±0.72) were significantly lower in silent NFATc1 + CD137 activated group than in the CD137 activated group (both P ＜ 0.05).The formation of the tube length ((1.28 ± 0.34) mm) and branch number (5.07 ± 0.35) were also significantly decreased in the CD137 inhibited group compared with the CD137 activated group (both P ＜ 0.05).Compared with the CD137 inhibited group,the formation of the tube length ((4.82 ± 0.09)mm) and branch number(24.44 ± 1.05) in the over expressed NFATc1 + CD137 inhibited group was increased (both P ＜ 0.05).Conclusion CD137 can promote the angiogenesis in atherosclerosis plaque by activating NFATc1.