0.05).There was no difference in the rate of PB1 between PMSG group and none PMSG group.In PMSG group,INH A solutions at concentrations of 200 ng/ml,350 ng/ml had higer rate of PB1 than that in control group(P

Objective To observe the effects of low molecular weight heparin (LMWH) combined with dexamethasone (Dex) on coagulation and fibrinolysis at the early stage in rats with acute respiratory distress syndrome (ARDS) induced by two-hit of oleic acid (OA) and lipopolysaccharide (LPS).Methods Forty healthy adult male Sprague-Dawley (SD) rats were randomly divided into sham operation, ARDS model, Dex, LMWH and combining therapy groups (8 rats in each group). The rat ARDS model was established by sequential two-hit with intravenous injection of OA 0.2 mL/kg in a tail vein and intra-peritoneal injection of LPS 5 mg/kg. After model establishment, the rats in Dex group were intra-peritoneally injected with 10 mg/kg Dex, and those in LMWH group were intravenously injected with 200 U/kg LMWH, while the rats in combining therapy group were given Dex and LMWH simultaneously with the same dosages and methods as above mentioned respectively. In sham operation group, however, the rats were intravenously injected with 0.4 mL of normal saline (NS) and were given 2 mL/kg of intra-peritoneal NS injection, and they accepted another 1 mL/kg NS intra-peritoneal injection 4 hours later, the other procedures being the same as those in the model group. The experiment was ended at 6 hours after the establishment of ARDS model. A light microscope was used to observe the pathological changes in lung tissues, partial pressure of arterial blood oxygen (PaO2) was measured, and the oxygenation index (PaO2/FiO2) was calculated. Wet to dry weight (W/D) ratio of lung tissues was also checked. Coagulation indexes were measured by solidification method, and the serum level of plasminogen activator inhibitor-1 (PAI-1) as well as the content of procollagen type Ⅲ (PC-Ⅲ) was determined by enzyme linked immunosorbent assay (ELISA).Results Under the light microscope, effusion of red blood cells, fibrin deposit in the lung interstitium and alveoli, formation of transparent membrane at alveolar wall, inflammatory cells infiltration in pulmonary interstitial tissue, and fibrinous thrombi in lung capillaries or lung arterioles were seen in the model group. Compared with model group, the red blood cells effusion and fibrin deposition in the lung interstitium and alveoli were less, inflammatory cells infiltration in pulmonary interstitium was alleviated and the fibrinous blood emboli in the lung capillaries or lung small arterioles were also decreased in Dex, LMWH and combining therapy groups, among the three groups, the best results being in the combining therapy group. Compared with sham operation group, the PaO2/FiO2 was significantly lowered [mmHg (1 mmHg = 0.133 kPa): 272.02±28.28 vs. 420.24±35.52,P < 0.01], the lung W/D ratio was obviously higher (5.59±0.40 vs. 3.82±0.28,P < 0.01), prothrombin time (PT) as well as thrombin time (TT) were markedly longer [PT (s): 18.78±1.57 vs. 16.36±0.97, TT (s): 39.02±5.03 vs. 29.22±8.83, bothP < 0.05], fibrinogen (Fib) content was significantly decreased (g/L: 1.82±0.26 vs. 2.69±0.40,P < 0.01), but both the serum contents of PAI-1 and PC-Ⅲ were remarkably elevated in model rats [PAI-1 (ng/L): 719.04±103.74 vs. 517.25±119.18, PC-Ⅲ (μg/L): 29.93±3.24 vs. 22.97±6.26, bothP < 0.01); Compared with model group, the level of PaO2/FiO2 was significantly elevated, and the lung W/D ratio was obviously decreased in Dex, LMWH and combining therapy groups respectively, the most significant changes being in combining therapy group [PaO2/FiO2 (mmHg): 376.78±25.25 vs. 272.02±28.28, lung W/D ratio: 4.14±0.42 vs. 5.59±0.4, bothP < 0.01] , in LMWH group, the prolongation of TT was the longest (s: 52.00±4.24 vs. 39.02±5.03,P < 0.05), while Fib contents in the three treatment groups were all obviously increased (g/L: 2.37±0.38, 2.59±0.51, 2.59±0.24 vs. 1.82±0.26,P < 0.05 orP < 0.01); meanwhile, the decrease of PAI-1 in combining therapy group was the greatest (ng/L: 546.02±93.94 vs. 719.04±103.74,P < 0.01). The indexes showed no statistically significant differences among the three treatment groups, except that PT in combining therapy group which was significantly longer than that in Dex and LMWH groups (s: 19.98±1.61 vs. 17.20±1.48, 17.02±2.34, bothP < 0.05). Conclusions The rats with ARDS induced by two-hit of OA combined with LPS have coagulation dysfunction and fibrinolytic inhibition. Using LMWH early can improve coagulation and fibrinolytic status in the rats with ARDS, and the therapeutic effects of LMWH plus Dex for treatment of ARDS are better than those of using each of them alone.

OBJECTIVE:To observe the effect of atorvastatin combined with methylprednisolone on the liver function of ne-phrotic syndrome patients. METHODS:The data of 93 patients with primary nephritic syndrome were retrospectively analyzed and divided into atorvastatin group,methylprednisolone group and combination group by different medication. Atorvastatin group was orally given atorvastatin 20 mg at bedtime,once a day+aspirin;methylprednisolone group was orally given methylprednisolone 0.8 mg/kg in the early morning,once a day+aspirin;combination group was given atorvastatin+methylprednisolone+aspirin(the same usage and dosage with the above-mentioned groups). The course was 4 weeks. The clinic data was observed,including ALT,AST, GGT,TB and DB before and after treatment,the incidence of patients with drug-induced liver disease and prognosis of patients with drug-induced liver disease. RESULTS:After treatment,the ALT,AST and GGT in atorvastatin group and combination group were significantly higher than before,with significant difference(P<0.05);compared with other parameters and all indexes in methylprednisolone group before and after treatment,there were no significant differences(P>0.05). There was no significant dif-ference in the elevated rate of ALT among groups(P>0.05);the incidence of drug-induced liver disease in combination group was significantly higher than atorvastatin group and methylprednisolone group,with significant difference(P<0.05). ALT in combina-tion group was significantly decreased and returned to pretreatment levels after atorvastatin withdrawal and 2 weeks of hepatoprotec-tants treatment for 7 patients with drug-induced liver disease. CONCLUSIONS:Atorvastatin combined with methylprednisolone has high risk on liver function in the treatment of nephrotic syndrome. Pretreatment levels can be recovered by both drug withdrawal and symptomatic treatment.

Objective To investigate clinical characteristics of moderate and severe ovarian hyperstimulation syndrome (OHSS) in assisted reproductive technique .Methods The clinical data of 179 cases with moderate and severe OHSS receiving in vitro fertili‐zation‐embryo transfer (including ICSI) in the hospital from June 2012 to April 2013 were analyzed retrospectively .According to the clinical characteristic ,the OHSS was classified as as the moderate type and severe type ,and the late type and early type . Results It was no statistics difference between moderate type and severe type in the patients age ,number of retrieved oocytes ,ad‐mission transaminase ,proportion of fibrinogen normal numbers(P> 0 .05) .But it was a statistics difference between moderate type and severe type in the occurring time days of hospitalization ,hematocrit on admission ,albumin value ,transaminase maximum ,albu‐min dosage used ,proportion of paracentesis number ,pregnancy rate(P < 0 .05) .It was no statistics difference between early type and late type in the patients age ,admission transaminase ,proportion of fibrinogen normal numbers(P> 0 .05) .But it was a statistics difference between early type and late type in the number of retrieved oocytes ,the proportion of moderate OHSS patients ,days of hospitalization ,hematocrit on admission ,albumin value ,transaminase maximum ,albumin dosage used ,proportion of paracentesis number ,pregnancy rate(P< 0 .05) .Conclusion Synthesizing OHSS patients′ blood indexes ,we can evaluated patients′ pathogenet‐ic condition ,the treatment of disease ,and took appropriate preventive measures as soon as possible .Patients with late type may be have more severe pathogenetic condition than patients with early type .

BACKGROUND: Hepatitis B virus (HBV) infection is one of the worldwide public health problem affecting about 300 million people. The envelope protein of HBV consists of three components known as preS1, preS2, and S antigen. According to the recent study, anti-HBs Ab showed effective neutralization ability against HBV from chronic hepatitis B and liver transplant patients, suggesting the possible development of therapeutic antibody. METHODS: Spleen cells immunized with S antigen of HBV were fused with myeloma cell line to obtain HBsAg specific monoclonal antibodies. High affinity antibodies against HBsAg (adr, ad and ay type) were selected by competitive ELISA method. Nucleotide sequence of the variable regions of monoclonal antibodies was analyzed by RT-PCR followed by conventional sequencing method. RESULTS: We produced 14 murine monoclonal antibodies which recognize S antigen of HBV. Two of them, A9-11 and C6-9 showed the highest affinity. The sequence analysis of A9-11 revealed that variable regions of the heavy chain and light chains are members of mouse heavy chain I (B) and light chain lambda 1, respectively. Likewise, the sequence analysis of C6-9 revealed that variable regions of the heavy chain and light chains are members of mouse heavy chain II (B) and light chain kappa 1, respectively. Neutralization assay showed that A9-11 and C6-9 effectively neutralize the HBV infection. CONCLUSION: These results suggest that A9-11 and C6-9 mouse monoclonal antibodies can be used for the development of therapeutic antibody for HBV infection.
Animals ; Antibodies ; Antibodies, Monoclonal* ; Base Sequence ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Hepatitis B Surface Antigens ; Hepatitis B virus* ; Hepatitis B* ; Hepatitis B, Chronic ; Hepatitis* ; Humans ; Liver ; Mice ; Public Health ; Sequence Analysis ; Spleen

OBJECTIVETo verify the existence and significance of calcium/calmodulin dependent serine protein kinase/inhibitors of differentiation 1 (CASK/Id1) pathway in fibroblasts of human keloid.

METHODSImmunofluorescence laser was used to confirm CASK and Id1 protein expression and localization in fibroblasts of the keloid and normal skin. RT-PCR and Western-blot were adopted to analysis the CASK and Id1 expression and differences between keloid and normal skin fibroblasts. The natural combination of CASK and Id1 protein of keloid fibroblasts was tested by immunoprecipitation.

RESULTSCASK and Id1 protein expression were both found in fibroblast cells of keloid and normal skin under normal circumstances. Most of CASK and Id1 were distributed in the cytoplasm and nucleus of fibroblasts. The results of RT-PCR showed that the expression of CASK mRNA in the keloid group was 0.658 +/- 0.024, which was lower than that in the normal control group (1.076 +/- 0.008, t = 11.159, P < 0.05). The expression of Id1 mRNA was 0.497 +/- 0.014, which was higher than that in the normal control group (0.307 +/- 0.017, t = 15.148, P < 0.05). The results of Western-blot showed that the expression level for CASK protein in the keloid group was 0.057 +/- 0.006, which was lower than that in the normal control group (0.168 +/- 0.012, t = 13.524, P < 0.05); the expression of Id1 protein was 0.812 +/- 0.035, which was higher than that in the normal control group (0.368 +/- 0.031, t = 16.356, P < 0.05). The results of immunoprecipitation showed that Id1 could be detected in the CASK precipitate, while CASK also could be detected in the Id1 precipitate. There was a natural binding of CASK and Id1 in keloid fibroblasts.

CONCLUSIONCASK/Id1 signal pathway may be existed and involved in the proliferation of keloid fibroblasts, which is related with the occurrence of keloid.

Cell Proliferation ; genetics ; Cyclin-Dependent Kinase Inhibitor Proteins ; genetics ; metabolism ; Fibroblasts ; metabolism ; Humans ; Inhibitor of Differentiation Protein 1 ; genetics ; metabolism ; Keloid ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Signal Transduction

The adsorption and activation of dimethyl carbonate on the surface of solid base were investigated by in situ FTIR, and the solid bases included magnesia, magnesium fluoride, Mg-Al mixed oxide and fluorine-modified Mg-Al mixed oxide. The FTIR results showed that dimethyl carbonate adsorbed on the surface of solid based by two modes of bidentate and unidentate complex. The bidentate was more active than the unidentate. Methoxyl group was formed from the adsorbed dimethyl carbonate on the surface of magnesia and Mg-Al mixed oxide. And fluomethyl group was formed from the adsorbed dimethyl carbonate on the surface of sodium fluoride. However, dimethyl carbonate on the surface of fluorine-modified Mg-Al mixed oxide showed preference for generating fluomethyl group. With the increasing of the treating temperature of samples, the methoxyl group was gradually formed on the surface. Accordingly, the fluorine-modified Mg-Al mixed oxide was found to be an excellent catalyst for methylation.

Objective:To investigate the effects of survivin inhibitor YM155{1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(pyrazin-2-ylmethyl)-4,9-dihydro-1H-naphtho[2,3-d] imidazolium bromide} on cell viability,apoptosis and Cysteinyl aspartate specific proteinase-3,Cysteinyl aspartate specific proteinase-8,Cysteinyl aspartate specific proteinase-9 of the thyroid carcinoma cell line B-CPAP in order to discuss mitochondrial mechanisms of apoptosis.Methods: B-CPAP cells were cultured in vitro and treated with YM155 at various concentrations(0,0.5,1,2,4,8 nmol/L)for 24,48 and 72h.The cell viability of B-CPAP cells were measured by CCK-8 assay.B-CPAP cells were randomly divided into 4 groups:B-CPAP cells were treated with YM155 at various concentrations(0,1,2 nmol/L)and 5 μmol/L Cisplatin(the positive control group)for 24 h.The effects of YM155 on B-CPAP cells apoptosis were evaluated by TUNEL and flow cytometry Annexin V-FITC/PI method.The expression level of Survivin and Caspase-3,Caspase-8 ,Caspase-9 were detected by Western blot analysis.Results: Compared with the 0 nmol/L group,YM155 significantly inhibited the cell viability of B-CPAP cells and induced their apoptosis (P<0.05 or P<0.01).Compared with the 0 nmol/L group,YM155 significantly reduced the expression level of Survivin and upregulated Caspase-3,Caspase-8 ,Caspase-9(P<0.05 or P<0.01).Conclusion: YM155 can inhibit the cell viability of B-CPAP cells and induce apoptosis,its possible mechanisms maybe related to upregulated expression level of Caspase-3,Caspase-8 and Caspase-9.

Objective To investigate the role of TLR4/NF-κB signaling pathway under the action of TAK-242 in the cardiomyocyte apoptosis after coronary micro-embolism (CME) in rats.Methods Fortyfive rats were randomized (random number) into three groups:sham operation,CME and CME plus TAK242 groups (n =15 per group).CME was induced by injecting polyethylene microspheres (42 μm) into the left ventricle except the sham group.CME plus TAK-242 group was treated with TAK-242 (2 mg/kg) via the tail vein of mice 30 min before CME modeling.Cardiac function was evaluated 6 h after operation.Tissue biopsy was stained with HBFP to measure the size of infarction area.TUNEL assay was used to detect cardiomyocyte apoptosis.Western blot and qPCR were used to evaluate the protein levels and mRNA expressions of TLR4,NF-κB p65 and cleaved caspase-3,respectively.Statistical analysis was performed using one-way analysis of variance followed by LSD-t test.Results Compared with the sham group,left ventricular ejection fraction (LVEF) in the CME group was significantly decreased [(68.91 ± 4.12) ％ vs.(84.80 ± 2.51) ％,P ＜ 0.05],and the infarction area (P ＜ 0.05),the apoptosis index [(3.36 ± 0.63) ％ vs.(0.19 ± 0.08) ％,P ＜0.05],the mRNA expressions of TLR4,NF-κB p65 and cleaved caspase-3 in CME group were increased significantly (all P ＜ 0.05).Compared with CME group,LVEF in the CME plus TAK-242 group was significantly improved [(75.58 ± 5.01) ％ vs.(68.91 ± 4.12) ％,P＜0.05],and the infarction area [(8.58 ± 2.12) ％ vs.(14.65 ± 4.23) ％,P＜0.05],the apoptosis index [(1.43 ± 0.51) ％ vs.(3.36 ± 0.63) ％,P ＜ 0.05],the mRNA expressions of TLR4,NF-κB p65 and cleaved caspase-3 in CME + TAK-242 group were decreased significantly (all P ＜ 0.05).Conclusions TAK-242 effectively improved CME-induced cardiac dysfunction by regulating TLR4/NF-κB signaling pathway and then reducing the cardiomyocyte apoptosis.

Objective:To explore the protective effect of vagus nerve stimulation (VNS) on the prognosis of rats suffering from cardiac arrest/cardiopulmonary resuscitation (CA/CPR) under different treatment timings.Methods:The method of percutaneous epicardial electrical stimulation was used to establish CA model of rat. Fifty-three male SD rats were randomly (random number) divided into the sham group ( n=5), CPR group ( n=12), PRE group ( n=12), POST5 group ( n=12) and POST30 group ( n=12). The sham group did not experience CA/CPR. VNS treatment was started at 30 min before CA (PRE group, n=12), 5 min after recovery of spontaneous circulation (ROSC) (POST5 group, n=12), and 30 min after ROSC (POST30 group, n=12) in different VNS-treated group, respectively. The electrical stimulation was applied to the vagus nerve for 30 min with a unified parameter. The neurological deficit scores at 24, 48, and 72 h after ROSC were recorded, and the survival rate in each group was observed. TUNEL staining was used to detect the apoptosis of cortical area and the expression of α7 nicotinic acetylcholine receptor (α7nAChR) in brain tissue was measured by immunofluorescence at 72 h after ROSC. Variables were compared with one-way analysis of variance, and survival for Kaplan-Meier curves were tested with the log-rank test. A P value less than 0.05 was considered statistically significant. Results:Compared with the CPR group (survival rate 33.33%), both pre-treatment (survival rate 75%) and post-treatment of VNS (POST5 group survival rate 75% and POST30 group survival rate 83.33%) significantly improved the 72 h survival rate after CPR ( P<0.05), mitigated neurological deficits after ROSC, reduced the positive rate of apoptosis neurons, and up-regulated the expression of α7nAChR in cerebral cortex. There was no significant difference among the VNS-treated groups (all P>0.05). Conclusions:Both pre-treatment and post-treatment of VNS can play a protective role in rats after CA/CPR, which may be related to the activation of α7nAChR-mediated anti-inflammatory and anti-apoptosis effects.