Objective To study the effect of glutathione(GSH)on liver antioxidative function of microcystin‐LR(MC‐LR)‐in‐duced mice .Methods Forty healthy clean class KM 5‐week old mice were selected and divided into five groups by the random sam‐pling method ,including the norml saline control group ,GSH control group ,MC‐LR group ,MC‐LR+low dose GSH group and MC‐LR+high dose GSH group ,8 cases in each group ,half male and half female .The experiment lasted for 15 d by intraperitoneal injec‐tion of MC‐LR ,then the liver histopathological changes ,liver tissue activity of superoxide dismutase (SOD) ,glutathione peroxidase (GSH‐Px)and content of malondialdehyde(MDA) were detected .Results Compared with the normal saline control group ,liver cell GSH level ,SOD and GSH‐Px activities in the MC‐LR group were significantly decreased (P<0 .05) ,while the MDA level was sig‐nificantly increased (P<0 .05) .Compared with the MC‐LR group ,the GSH level ,SOD and GSH‐Px activities in the MC‐LR+low dose GSH group and MC‐LR+high dose GSH group were significantly increased (P<0 .05) ,while the MDA level was significant‐ly decreased(PP<0 .05) .Conclusion The GSH intervention can alleviate MC‐LR induced mouse liver oxidative toxicity and has protective effect on the liver to some extent .

Objective To explore the structure of ribosomal protein S7 ( RPS7) gene of giant panda ( Ailuropoda melanoleuca) and investigate its homologies with other already reported sequences,including Homo sapiens,Mus musculus,Rattus norvegicus and Bos taurus. Methods The cDNA of RPS7 was cloned from the giant panda by RT-PCR. The sequence data were analyzed by GenScan software. Blast 2. 1 was used to study the homology of the obtained RPS7 sequence with the gene sequences of other species; Open reading frame ( ORF) of the DNA sequence was searched using ORF finder software; Protein structure of the RPS7 sequence cloned was deduced using Predict Protein software. Results The full length of the sequence fragment was 589 bp containing an ORF of 585 bp. The deduced protein sequence showed that the protein was composed of 194 ami- no acids and its estimated molecular weight was 22. 126 85 ?103 with a pI of 10. 09. There were 7 different pat- terns of functional sites: one N-glycosylation site; two cAMP and cGMP-dependent kinase phosphorylation sites; four casein kinase C phosphorylation sites; one casein kinase Ⅱ phosphorylation site; two N-myristoylation sites; two amidation sites and one ribosomal protein S7e signature site in the RPS7 protein. Further analysis indicated that the sequence of RPS7 and the protein encoded were highly homologous to some mammals reported.Conclusion The complete coding sequence of RPS7 gene has been cloned through RT-PCR successfully, which is the first report on the RPS7 gene from the giant panda.

OBJECTIVE To analyze the pathogens of severe pneumonia induced by different underlying diseases in(hospital) and to evaluate the difference of the pathogens to antimicrobial susceptibility test.METHODS The(bacteria) through sputum culture with VITEK-AMS,and the antimicrobial susceptibility against(bacteria) by K-B method were tested in hospital from 2002 to 2005.RESULTS There were 106 patients with 173 strains of isolated(pathogenic) bacteria,including Klebsiella pneumoniae(KPA) 20.9%,and Escherichia coli(ECO)(16.3%) from the aspiration severe pneumonia;Pseudomonas alcaligenes(PAL) 18.9%,P.aeruginosa(PAL)(16.2%) and Staphylococcus aureus(SAU) 16.2% from the obstructive severe pneumonia;KPN 22.6%,SAU(18.9%),and PAE 17.0% from the COPD complicated with severe pneumonia and PAE 25.0% and SAU 20% from the(hospital-)acquired pneumonia(HAP).CONCLUSIONS The constructed ratio of pathogens is different between(severe) pneumonia infected by different underlying diseases and community-acquired pneumonia without any(underlying) disease.The resistant pathogens are increasing significantly in cases with HAP.

Objective To explore the value of placental alpha-microglobulin-1 in vagina liquid to diagnose premature rupture of membranes. Methods A prospective observational study to initial evaluation included both the standard clinical evaluation for rupture of membranes and placental alpha-microglobulin-1 immunoassay. Rupture of membranes was diagnosed if fluid was seen leaking from the cervical os or if two of the following three conditions were present: pooling of fluid, positive nitrazine test, or feming. Rupture of membranes was diagnosed definitively on review of the medical records after delivery. Results Placental alpha-microglobulin-1 immunoassay confirmed rupture of membranes at initial presentation with a sensitivity of 100% (89/89), specificity of 91% (10/11), positive predictive value of 99% (89/90), and negative predictive value of 100% (10/10),false positive rate of 9% (1/11). Placental alpha-nricroglobulin-1 immunoassay was better than the conventional clinical assessment in confirming the diagnosis of rupture ofmembranes (P<0.01). Conclusion Measurement of placental alpha-microglobulin-1 in cervicovaginal secretions is superior to conventional clinical assessment in the diagnosis of rupture of membranes.

Objective To observe the effects of tetramethylpyrazine on intraoperative autotransfusion of blood quality , analysis of ligustrazine in autotransfusion of clinical value of blood quality control.Methods 68 patients undergoing elective surgery in our hospital from February 2015 to May 2016 were selected as the research object,and were randomly divided into control group and experimental group,each group of 34 cases.Two groups of patients in the intraoperative autologous blood transfusion and the control group according to conventional autologous transfusion in operation ,test group of patients with intravenous injection of ligustrazine injection in the recovery of blood before adding ligustrazine injection in the recovery process of blood.The two groups were collected before blood transfusion,observe the morphology of red blood cells, red blood cell fragments under the microscope and compared;the two groups were collected after reinfusion of blood two hours,one and five days in peripheral venous blood, the determination of superoxide dismutase (SOD), malondialdehyde (MDA), T lymphocyte subsets level of two groups were compared before and after one and five days. Results The erythrocyte deformability and erythrocyte debris of the two groups were higher than those before operation (P<0.05).The blood red blood cell deformability and the number of red blood cell debris in the blood transfusion group were significantly higher than those in the control group Significantly lower than the control group (P<0.05).The levels of SOD were significantly lower at two hours and one day after operation than those before operation, and the serum SOD was still lower than that before operation in the control group,The SOD in the experimental group was significantly higher at two hours, one and five days in the control group,MDA was lower than the control group(P<0.05).The CD3 +,CD4 +, CD4 +/CD8 +levels were significantly lower at two hours and one day after operation than those before operation (P<0.05).CD3 +,CD4 +and CD4 +/CD8 +in the control group were still lower than those in the control group at five days after operation(P<0.05),the levels of CD3 +, CD4 +, CD4 +/CD8 +were significantly higher in the experimental group than those in the control group at two hours,one and five days after treatmen (P <0.05).There was no significant difference between the two groups in the incidence of adverse reactions.Conclusion The addition of preoperative intravenous injection of ligustrazine,blood recovery process,can effectively protect the transfusion of red blood cell integrity,reduce the effect of transfusion of blood and blood antioxidant capacity in patients with T lymphocyte immune function ,to improve the quality of blood transfusion patients has important clinical value .

Background and purpose:Selenium is one of the essential trace elements for human activities, and plays an incomparable role in maintaining human health. It was reported that selenium compound 1,4-bis[2-(benzylse-lanyl)ethoxy] (BSEA) anthracene has antiseptic and antiphlogistic effects. However, the mechanisms underlying anti-cancer effects of BSEA are rarely reported. BSEA-induced apoptosis in human laryngeal carcinoma Hep-2 cells and its mechanisms were studied.Methods:Methyl thiazolyl tetrazolium (MTT) assay was used to determine inhibition ratio of Hep-2 cells 24 hours after Hep-2 cells were treated with different concentrations of BSEA. Fluorescence microscope was used to observe the morphology change of apoptosis in Hep-2 cells. The apoptosis was detected by Annexin Ⅴ-FITC. Mi-tochondrial membrane potential was assayed by JC-1. Microplate reader detected the activity of caspase-3 and caspase-8. The mRNA and protein levels of Bax and XIAP were measured by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot.Results:The results showed that BSEA caused a dose-dependent inhibition of the growth of human laryngeal carcinoma cell line Hep-2in vitro, andIC50 was 35.74μmol/L. The apoptotic bodies were distinctly observed at a concentration of 80μmol/L of BSEA by AO fluorescence staining. This study found that the eversion of phosphatidyl serine intensified, and mitochondrial membrane potential also began to decline. The activity of caspase-3 appeared the tendency of dependence on dosage, while the activity of caspase-8 did not change significantly. The mRNA and protein expression level of Bax increased, whereas the mRNA and protein expression level of XIAP de-creased.Conclusion:Therefore, BSEA could obviously inhibit human laryngeal carcinoma Hep-2 cells proliferation and induce apoptosis via the mitochondrial pathway.

Objective To investigate the effect of continuous veno-venous hemofiltration (CVVH) on pulmonary vascular permeability in the early stage of septic shock.Methods Fifty-one patients of septic shock admitted in intensive care unit of the First Affiliated Hopsital of Medical College,Zhejiang University between June 2010 and December 2011 were randomized into two groups by simple random method.Routine treatment was carried out to patients in group A,and routine treatment plus CVVH to patients in group B in the first 72 hours.Repeated respiratory mechanic hemodynamic measurements were done at baseline (Tb) before treatment,at 48 hours (T48) and 72 hours (T72) after the treatment.Pulmonary compliance (Cst),platform pressure (Pplat),oxygenation index (PaO2/FiO2),extravascular lung water index (EVLWI) and E-selectin determined at each time point were recorded in two groups.Results (1) Pplat were decreased significantly after treatment in two groups,and the Pplat in group B were all lower than those in group A at T48 and T72 (t =2.215,P ＜ 0.05 ; t =4.266,P ＜ 0.01).Cst were elevated after treatment in two groups,and Cst in group B were all higher than those in group A at T48 and T72 (t =2.516,P ＜0.05 ; t =3.052,P ＜ 0.01).(2) Compared to before treatment,PaO2/FiO2 increased significantly after treatment in two groups,and PaO2/FiO2 in group B were all higher than those in group A at T48 and T72 (t =2.732,P ＜0.01 ; t =3.511,P ＜0.01).(3) EVLWI were decreased significantly after treatment in two groups,and the EVLWI in group B were all lower than those in group A at T48 and T72 (t =2.597,P ＜0.05; t =2.125,P ＜ 0.05).ITBVI,CI,SVRI and MAP did not change over time in two groups compared with those at Tb (all P ＞ 0.05).(4) E-selectin were decreased significantly after treatment in two groups,and the E-selectin in group B were all lower than those in group A at T48 and T72 (t =2.154,P ＜0.05 ; t =3.581,P ＜ 0.01).Conclusion CVVH in early stage of septic shock can improve pulmonary vascular permeability and oxygenation,increase Cst,and decline EVLWI,with neither increased hemodynamics.

Objective To investigate the effects of biatrial infusion on pulmonary artery pressure (PAP)after cardiopulmonary bypass (CPB) in patients undergoing mitral valve replacement.Methods Twenty NYHA Ⅱ or Ⅲ patients aged 22-53 yr weighing 34-57 kg undergoing mitral valve replacement complicated by pulmonary hypertension (mean pulmonary artery pressure (MPAP) ＞ 50 mm Hg) were randomly divided into 2 groups ( n = 10 each): infusion via right atrium group (group R) and infusion via both atria group (group B). After induction of anesthesia, a three cavity floating Swan-Ganz catheter was placed via right internal jugular vein to monitor CVP,PAP, pulmonary capillary wedge pressure (PCWP) and CO. The patients received infusion of prostaglandin E1 30-150 ng· kg- 1 · min - 1 and phenylephrine 0.2-0.6 μg· kg- 1 · min- 1 via central veins in group R and infusion of prostaglandin E1 30-150 ng·kg-1 ·min-1 via central veins and phenylephrine 0.2-0.6 μg·kg-1 ·min-1 via left atrium in group B. MAP, HR, MPAP, PCWP, CVP and CO were recorded 5 min before administration (T0), and 5,10, 30 and 60 min after administration (T1-4). Pulmonary vascular resistance index (PVRI), systemic vascular resistance index (SVRI) and CI were also calculated. Results D:\1111111111\MDB\zhmzxzz98201008.mdbCompared with the value at T0, MAP, MPAP, PCWP and PVRI were significantly decreased, while CI was increased at T1-4 in group R, and MAP, CI and SVRI were significantly increased, while HR, MPAP, PCWP, CVP and PVRI decreased at T1-4 in group B ( P ＜ 0.05).MAP, CI and SVRI were significantly higher, while HR, MPAP, PCWP, PVRI and CVP lower in group B than in group R ( P ＜ 0.05). Conclusion Biatrial infusion can effectively reduce PAP and pulmonary vascular resistance after CPB in patients undergoing mitral valve replacement.

Objective To investigate the way that nasopharyngeal carcinoma (NPC) and NPC stem cells develops resistance to cisplatin through anti-reactive oxygen species mechanism. Methods Using CCK-8 cell counting kit, we measured the half inhibitory concentration of cisplatin against NPC cellsCNE-2and NPC stem cellsCNE-2S, and compared their resistant index. We examined the differences in the reactive oxygen species (ROS) levels, total glutathi?one (GSH) levels, and total superoxide dismutase (SOD) levels between CNE-2 and CNE-2S at different concentrations of cisplatin administration (0.1,0.5 and 1.0μmol·L-1). Using q-PCR, we determined the mRNA expression level of GSS, GCLC, GCLM, SOD1 and SOD2 after 48 hours administration of cisplatin at 1 μmol · L-1. Protein expression level of SOD2 was also tested using Western Blot after 48 hours administration of cisplatin at 1μmol · L-1. Upon silencing the SOD2 in NPC cell through siRNA, Trypan blue was used to analyze cell survival after cisplatin was administrated at 1μmol · L-1. Results The inhibition concentration of cisplatin against CNE-2 was higher than that against CNE-2S (μmol · L-1:9.8 ± 1.1 vs 2.4 ± 0.6,P<0.05). ROS levels in CNE-2 and CNE-2S both rise with cisplatin administration, but ROS levels of CNE-2 before and after cisplatin treatment were both higher than those in CNE-2S (P<0.05). The total gluta?thione levels in CNE-2 and CNE-2S were both increased after 1μmol·L-1 cisplatin treatment but there is no significant dif?ference in levels of glutathione between these two cell lines. After treated with cisplatin, SOD level were increased in both CNE-2S and CNE-2, but it is higher in CNE-2S than that in CNE-2 (P<0.05). The mRNA levels of GSS, GCLC, GCLM, and SOD1 were not different significantly between in CNE-2 and in CNE-2S with or without cisplatin treatment. However, SOD2 in CNE-2S were higher than that in CNE-2 on both mRNA and protein levels (P<0.05). Silenced SOD2 disrupted the resistance of cisplatin in CNE-2S. Conclusion These data suggest that NPC stem cells (CNE-2S) enhance its drug re?sistance to cisplatin through highly expression of SOD2 which posed anti-ROS capacity.

Background and purpose: The Writ cell-signaling pathway is the key cellular developmental pathway. Dysregulation of this pathway has been implicated in the initiation and progression of cancer. Adenomatous polyposis coli (APC) is an important tumor suppressor gene of the Writ signaling pathway. The methylation of APC promoter and the accompanying loss of the APC transcript have been shown to occur in a significant proportion of cancers. However, there are few reports on the relationship between cervical cancer and methylation of APC. This study was aimed to investigate the promoter methylation status of the APC genes in cervical cancer and its correlation between clinicopathologic characteristics and the infection of high-risk HPV DNA. Methods: Promoter methylation was evaluated using a MSP (methylation-specific polymerase chain reaction) in 95 cervical cancer tissue specimens and 20 normal controls. The relationship between clinicopathologic parameters and the methylation status was evaluated. Results: The frequencies of promoter methylation of APC in cervical cancer were 56.8%. Cervical cancer had significant higher methylation frequencies than that of the controls (10%, P<0.01). The result showed that the methylation analysis of APC promoter and high-risk HPV DNA testing had good consistency (Kappa=0.348, P<0.001). The promoter methylation of APC was significantly higher in adenocarcinoma (AC) than in squamous cell carcinoma (SCC) (74.1% vs 50.0%, respectively, P<0.05). The larger tumor size, positive lymph node metastasis and positive HPV DNA exhibited an increased promoter methylation frequency for APC (P<0.05). There were no significant associations between the methylation frequencies for APC gene to age, invasion depth, FIGO stage and histological grade. Conclusion: Our results suggested that the promoter methyiation of APC and high-risk HPV DNA testing had good consistency. APC gene promoter methylation was a frequent epigenetic event in cervical carcinoma and had a significant correlation with cancer pathological types.