Objective To confirm the best dry technics,the colormetry of anthrone-sulfuric method was employed to determine the content of polysaccarides in fresh water mussel and three factors of temperature,dry time and tiled destiny were compared.Method Colorimetry of anthrone-sulfuric acid.Results and ConclusionThe best dry technics was: 2.5 kg/m2 tiled destiny,temperature at 90 ℃ and drying within 2 hours.The content of total amylase in polysaccharides in fresh water mussel was 21%.The method appeared to be reliable and operable for quality control.

Objective To investigate the effects of amyloid-β (Aβ)25-35 on endoplasmic reticulum (ER) stress and apoptosis in cultured rat cardiomocytes,and to elucidate the role of ER stress in the injury of cardiomocytes induced by Aβ25-35.Methods The isolated rat myocardial cells were cultured in vitro.Following stimulation of Aβ25-35 with different dose,the survival ratio was observed with methyl thiazolyl tetrazolium (MTT) method.Hoechst33258 staining was used to observe the morphology of apoptotic changes.The percentage of apoptotic cardiomyocytes was quantified with flow cytometry.The expressions of ER stree proteins,including X box-binding protein-1 (XBP-1),glucose-regulated protein 78 (GRP78),and CCAAT/enhancer-binding protein homologous protein (CHOP) were measured with Western blot.The cleaved caspase-3 and cleaved poly (ADP-ribose) polymerase (PARP) were measured with Western blot.Results Aβ25-35 decreased the survival ratio and induced the apoptosis of cultured rat cardiomocytes in dose-dependent mode.Meanwhile,Aβ25-35 increased the expressions of ER stree proteins,including XBP-1,GRP78,and CHOP.Aβ25-35 increased the expressions of cleaved caspase-3 and cleaved PARP.Conclusions Aβ25-35 could induce the apoptosis of rat cardiomyocytes,which were involved in ER stress possibly.This study might provide a new strategy for clinical treatment of Alzheimer's disease (AD)-associated myocardial injury.

Objective To investigate the characteristics of CD8+ stem memory T cells (CD8+Tscm) in patients with chronic HIV-1 infection before and after antiretroviral therapy (ART) and to analyze their associations with progression of HIV-1 infection.Methods Thirty-six patients with chronic HIV-1 infection and 20 healthy subjects were enrolled in this study.Flow cytometry was performed to detect the percentages and absolute numbers of CD8+Tscm in patients with chronic HIV-1 infection before and after antiretroviral therapy (ART) as well as in healthy subjects.Correlation analysis was used to demonstrate the relationships between CD8+Tscm and markers for progression of HIV-1 infection (CD4+T cell count, HIV-1 viral load and level of activated T cells).Results The percentages and the absolute numbers of CD8+Tscm in patients with chronic HIV-1 infection had no significant change before and after ART.They were respectively positively correlated with the percentages and the absolute numbers of CD4+Tscm.The percentage of CD8+Tscm was proportional to the percentage of CD8+ central memory T cells (CD8+Tcm), but was inversely proportional to the percentage of CD8+ effector memory T cells (CD8+Tem).In addition, the percentages of CD8+Tscm in patients with HIV-1 infection were negatively correlated with the viral loads before ART.Conclusion CD8+Tscm are responsible for maintaining the homeostasis of other CD8+T cell subsets.CD8+Tscm play an important role in inhibiting viral replication.

Objective To clone and express bradyzoite antigen 1(BAG1) gene of T. gondii,and analyze the immunoreactivity of the recombinant product. Methods The differentiation of T. gondii RH strain tachyzoites into bradyzoites was induced in vitro,and the coding sequence of BAG1 was amplified from bradyzoites by RT-PCR. The PCR product was analyzed by sequencing. The BAG1 coding sequence was further subcloned into the plasmid pET32a(+). The plasmid pET32a(+)-BAG1 was then transformed into BL21(DE3) to express after IPTG induction. The expression product was purified with Ni-NTA agarose and the purified BAG1 was further analyzed by Western blotting and ELISA. Results BAG1 cDNA was amplified from bradyzoites. After IPTG induction,BAG1 was expressed in a fusional form in E. coli. Western blotting showed that the purified recombinant protein could be specifically recognized by sera from mice chronically infected by T. gondii B36 strain. ELISA showed that the positive rate of T. gondii IgG antibodies of 350 human sera detected by the recombinant BAG1(17.4%) was higher than by recombinant SAG1 (12.6%)(P

BACKGROUND: Besides being a basic growth factor crucial to maintain and promote the development, differentiation and survival of the central nervous system, nerve growth factor(NGF) also plays an important role in the repair of injured peripheral nerves.OBJECTIVE: To investigate the effect of the muscular injection of NGF on the regeneration and functional recovery of rat sciatic nerve after crush injury.DESIGN: A randomized controlled pilot study in rats with repeated observation and measurement.SETTING: Center for new drug evaluation in a military medical university.MATERIALS: This study was performed in the Center for New Drug Evaluation, Department of Basic Medicine, Second Military Medical University during the period from July 1999 to March 2000, using 40 SD rats weighing 200 to 250 g(of either sex of half number) provided by the Sino-British SIPPR/BK Lab Aninal Ltd (Shanghai).METHODS: Forty rats were randomized into high-, mid- and low-dose NGF treatment groups, normal control group and model control group. The sciatic nerves were clamped at 6 nm distal to the sciatic notch to induce a 4-mm-wide area of crush injury. In the high-, mid-; and low-dose NGF groups, the rats were given NGF at 8, 4 and 2 μg/kg per day(corresponding to 1.6 × 10 3, 8 × 10 2 and 4 × 10 2 IU/kg per day) respectively via the muscular injection for 56 consecutive days.(NCVs) and sciatic function index(SFI) at different time points after the RESULTS: Compared with that of the model control group, the NCVs significantly increased in the high-dose NGF group 35 and 56 days after the injury,and in the mid-dose NGFgroup at 35 days(t=2.32-5.14, P ＜0.05-0.01 ). The SFIs significantly increased in all NGF-treated groups at 14 days ( t = 2. 29-6.28, P ＜ 0.05-0.01 ), with the recovery most conspicuous in high-dose NGF group; No significant difference in the SFIs was found between the NGF-treated groups on the 56th day. Morphological examination of the tissues identified no significant difference in the nerve myelin sheaths and axons in NGF-treated groups as compared with the normal control group,while in the model control group, myelin sheath dislocation with unclear microstructure was observed, accompanied by Schwann cell degeneration and necrosis.CONCLUSION: NGF promotes the repair of the damaged nerve myelin sheath and axon and stimulates nerve fiber regeneration and function recovery of the crushed rat sciatic nerves.

Objective To clarify the mechanism of Yifeiqinghua granules on tumor angiogenesis and tumor inhibition by observing its influence on the expression of vascular endothelial growth factor (VEGF) and its receptor KDR in Lewis lung cancer mice. Methods Eighty C57BL/6 inbred mice subcutaneously planted with Lewis tumor cell suspension 0.2 mL were randomly divided into 8 groups:blank group, control group, low-, medium- and high-dose of Yifeiqinghua granules groups, Gefitinib group, Gefitinib plus medium-dose of Yifeiqinghua granules group and CTX group, 10 mice in each group. Drugs were administrated from the second day. CTX group was administered on third day and seventh day, other groups were administered for 14 days. The mice were killed to take the tumor on the 15th days. VEGF and KDR expression in tumor tissue was measured by Western Blot. Results Compared with the blank group, the expression of VEGF was significantly decreased in the low- and medium-dose of Yifeiqinghua granules groups (P<0.01). Gefitinib group’s VEGF value was lower than the blank group (P<0.05). Compared with the blank group, the expression of VEGF was significantly decreased in the high-dose of Yifeiqinghua granular group, Gefitinib plus medium-dose of Yifeiqinghua granules group and CTX group (P<0.001). Compared with the blank group, the expression of KDR was significantly decreased in the low-, medium- and high-dose of Yifeiqinghua granules groups, Gefitinib group and CTX group (P<0.05). Conclusions The possible mechanism of Yifeiqinghua granules in treating non-small cell lung cancer is reducing the expression of VEGF and KDR, thus play an inhibitory effect.

Objective To observe the acute skin and mucous membrane reactions in patients treated with concurrent radiochemotherapy for nasopharyngeal carcinoma,and to analyze the influencing factors.Methods A total of 85 nasopharyngeal carcinoma cases treated with concurrent radiochemotherapy were enrolled in the study.Fifteen clinical and laboratory indexes,including BMI,radiation dose,degree of acute oral mucous and skin reactions and blood routine test were observed weekly.Univariate and multivariate regression analysis were performed to assess the factors,and screen the independent factors.Results Multiple-factor analysis showed that the risk factors cloesly related with acute radioactive oral mucosa reactions were smoking history(OR =3.467,P ＜ 0.05),single-dose of gross tumor volume (GTV) ＞2.15 Gy(OR =3.393,P ＜ 0.05),while those with acute radiation skin reactions were diabetes history(OR =87.859,P ＜ 0.05) and hemoglobin values 1 week before radiotherapy ＞ 130 g/L (OR =21.404,P ＜ 0.05).Conclusions In the patients treated with concurrent radiochemotherapy for nasopharyngeal carcinoma,smoking history and single-dose of GTVnx is the independent risk factors of acute radiation oral mucosa reactions,while diabetes history and hemoglobin values I week before radiotherapy are the independent factors of acute skin reactions.

Objective To understand the change of transient bacteria on surface in clinical ward before and after terminal disin‐fection ,provide the basis for controlling of hospital infection .Methods Surface samples were collected before and after terminal dis‐infection in infected patch of our hospital ,and then bacterial in the samples were cultured and identified .Compared changes about number and type of samples bacterial ,distribution of common clinical pathogenic bacteria before and after of the terminal disinfec‐tion .Results The surface colony number < 10 CFU /cm2 accounted for 63 .54% after terminal disinfection ,compared with the dis‐infection before 56 .29% ,increased 7 .25 percentage points .Surface sampling microorganism detecting rate decreased by 6 .74% . Surface average bacteria colony had different degree decreased before and after disinfection ,except the bed frame and quilt cover . Water tap ,which was the largest amount of bacteria surface ,followed by the bedside table .Before and after disinfection ,the mainly common microorganism was environment bacteria in infected patch ,including coagulase negative staphylococcus ,gram positive ba‐cilli ,Micrococcus ,Acinetobacter spp .Clinical common pathogenic bacteria mainly isolated from the department of brain surgery (9 .49% ) ,department of hepatology(8 .76% ) ,department of dermatology (8 .76% ) ,department of pediatrics (8 .03% ) ,emergency department (7 .30% ) .Pathogenic bacteria living areas were mainly the bedside table (21 .17 % ) ,water tap (18 .25% ) ,bed rest (12 .41% ) .Conclusion Terminal disinfection could effectively reduce the number of bacteria in the infected patch ,improve the ward environmental sanitation quality ,it have an important significance in the prevention of hospital infection control .

Objective To observe the effects of berberine on ICAM-1, VCAM-1 expression and inflammatory cells exudation in mice with viral pneumonia caused by influenza virus, and explore its anti-injury effect. Methods Totally 108 BALB/c mice were randomly divided into control group, model group, and berberine group. 25 μL 50 LD50 influenza virus, mouse lung-adapted strain, was intranasally inoculated to model group and berberine group. 1 h after infection, control and model group were intragastrically given 25 μL distilled water, berberine group was treated by intraperitoneal injection with berberine at a dose of 0.005 g/(kg·d) for 5 days, twice per day. On day 2, 4 and 6 after infection, immunocytochemical method was used to detect ICAM-1 and VCAM-1 expression, and sorting cell count of leukocytes in bronchoalveolar lavage fluid (BALF) were counted. Results The expression of ICAM-1 and VCAM-1 in model group increased obviously on day 2, 4, 6, and which in berberine group decreased compared with model group (P<0.01). WBC, mononuclear cell, eosinophile cell and neutrophil cell number in model group increased significantly. WBC and neutrophil cell number decreased in berberine group on day 6 (P<0.01), and the mononuclear cell number decreased on day 4 (P<0.01). Conclusion Berberine inhibited the expression of ICAM-1 and VCAM-1, and decreased the inflammatory cells exudation in lung of mice with viral pneumonia caused by influenza virus. Berberine has protective effect on inflammatory injury of lung tissue in mice with viral pneumonia caused by influenza virus.

Objective To summarize the application of ATP bioluminescence assay in surveillance of terminal disinfection of effects ,so as to provide the basis for intervention of disinfected effects .Methods ATP bioluminescence assay were employed to randomly test the surfaces of operating objects in therapeutic rooms and beside tables in wards ,total 144 object surfaces ,of each clinical departments in the whole hospital .The values of ATP bioluminescence assay were read on‐site ,0-250 RLU was recognized as qualification ,while disqualification when >250 RLU .The disqualified object surfaces were performed on‐site intervention that all of them were re‐disinfected ,the results were compared .Results Both the surfaces of operating objects and beside tables were dis‐qualified before disinfection ,and the values of ATP bioluminescence assay were 780 ± 10 .34 RL and 853 ± 13 .29 RLU respectively . The pass rates of ATP bioluminescence assay was 61 .97% of operating surfaces and 79 .45% of beside table surfaces the first dis‐infection .The disqualified sites were retested following on‐site intervention .The values of ATP bioluminescence assay were 431 .02 ± 0 .53 before intervention and 1 .43 ± 0 .59 after intervention ,and the difference was statistically significant .Conclusion ATP bi‐oluminescence assay can get more immediately ,simple and timesaving in evaluating the effect of disinfection and estimate the effi‐ciency of disinfection timely ,which can also provide the scientific basis on on‐site intervention so as to improve the execution power of hospital infection management .