Objective We want to study the therapeutic efficiency of autologous ADMSC transplantation in myocardial infarction.And we try to find out a good way to improve the therapeutic efficiency by using the combination of gene therapy and cell therapy.Anti-apoptotic protein XIAP was selected to fight against the ischemic environment of myocardic infaction.Methods ADMSC was isolated from rat inguinal fat tissue.ADMSC was cultivated with DMEM.XIAP experession plasmid was elertco-transduced to ADMSC.The anti-apototic function of XIAP was tested by serum stavation induced apotosis.The method of ligation of the left anterior descending artery was used to prepare the Myocardial infarction model.Then rats were randomly separated into three groups to receive direct epicardial injections of normal saline,or ADMSCs cell suspension or XIAP modified ADMSCs cell suspension at five sites in central zones of myocardial infarction and border zone.Cardiac function and the infarct size were evaluated 4 weeks after ADMSCs transplantation.Results West blotting suggest that,XIAP over-expression block serum starvation induced apotosis.It showed that there are significant statistic difference among XIAP modified ADMSC transplantation group,ADMSC transplantation group and control group 4 weeks after myocardial infarction (P ＜ 0.05).Left ventricular ejection fraction(LVEF) showed a significant improvement in ADMSCs transplantation group compared to control group (P ＜0.05).Left ventricular end systolic diameter(LVDs) and left ventricular end diastolic diameter(LVDd) of ADMSCs group were smaller than control group(P ＜ 0.05).The area of myocardial infarction was significantly reduced in the ADMSCs transplantation group compared to the saline group(P ＜0.05).Compared to ADMSCs transplantation group,effect of the XIAP modified ADMSC in rats with myocardial infarction is more obvious.The reduction of LVEF of XIAP modified ADMSCs group was signific antly lower(9％) than the ADMSCs group(16％) (P ＜ 0.01).Infarction area in XIAP modified ADMSCs group(3.26 ±0.95)％ was smaller than ADMSCs group(5.17 ±2.03)％ (P ＜0.05).Conclusion Autologus ADMSC transplantation is an efficient therapeutic tool in myocardial infarction therapy.Over expression of XIAP can partly inhibit lowserum induced apotosis of ADMSC in vitro,and it can improve left ventricular function better in vivo.Over expression XIAP of ADMSC can improve the therapeutic efficiency compare to ADMSC transplantation.

Objective To analyze the efficacy of rosuvastatin on the patients with hyperlipidemia and hypertension.Methods From March 2011 to June 2012,112 cases with hyperlipidemia and hypertension in our hospital were enrolled in this study.Patients were randomly divided into treatment group and control group (56 patients,each).Patients in control group were treated with oral amlodipine 5 mg/d.Patients in treatment group were treated with oral rosuvastain 10 mg/d and oral amlodipine 5 mg/d.One month after the treatment,the levels of blood pressure,total cholesterol (TC),tryglyceride (TG),low density liporotein (LDL-C),high density lipoprotein cholesterol (HDL-C),high sensitivity C-reactive protein (hsCRP) were determined.The occurrence of adverse effects were observed.Results One month after treatment,systolic blood pressure and diastolic blood pressure were significantly decreased in both two groups compared with pre-treatment [Control group:(135.2±9.51)mm Hgvs.(59.2±7.3)mm Hg,(88.8±5.2)mm Hg vs.(99.5±8.3)mm Hg,t=4.95,2.87; Treatment group:(130.2±5.5)mm Hg vs.(160.3±9.3)mm Hg,(86.7± 10.2)mm Hg vs.(99.7±8.3)mm Hg,t=5.03,2.94,all P＜0.01],but more declines were found in treatment group than in control group(t=3.96,3.42,both P＜0.001).The levels of LDL-C,TG and TC were significantly decreased in both two groups compared with pre-treatment [Control group:(2.64±0.72)mmol/L vs.(3.97±0.84)mmol/L,(1.89±0.25)mmol/L vs.(2.56±0.45)mmol/L,(4.23±0.56)mmol/L vs.(7.36±0.48)mmol/L,t=2.58,3.03,2.36,P=0.013,0.004,0.022;Treatment group:(1.75 ± 0.68) mmol/L vs.(3.85 ± 0.79) mmol/L,(1.71 ± 0.18) mmol/L vs.(2.63±0.42)mmol/L,(3.18±0.47)mmol/L vs.(7.20±0.56)mmol/L,t=2.77,3.16,2.59,P=0.008,0.003,0.012,respectively],but more declines were observed in treatment group than in control group(t=6.73,4.37,10.70 respectively,all P＜0.05).The HDL-C concentrations were increased in both two groups compared with pre-treatment [Control group:(0.97±0.26)mmol/L vs.(0.75±0.31)mmol/L,t=2.89,P=0.006; Treatment group:(1.09±0.23)mmol/L vs.(0.72±0.24)mmol/L,t=3.01,P=0.004],but more increment were observed in treatment group than in control group(t=2.59,P＜0.05).The hsCRP concentration was significantly reduced in treatment group compared with pre-treatment [(1.32±0.17) mg/L vs.(4.97±0.13) mg/L,t=4.40,P＜0.001].There were no significant differences in liver and kidney function between the two groups.Serious adverse effects were not found.Conclusions Rosuvastatin combined with routine antihypertensive therapy can effectively decrease the levels of serum LDL-C,TG,hsCRP; increase serum HDL-C concentration and blood pressure can be effectively controlled.

Objective To study the relationship of different histological types and degrees of infiltration of gastric adenocarcinoma on lymph node metastasis.Methods The clinical data of 1 615 cases of gastric adenocarcinoma were analyzed retrospectively,and the rate and number of lymph node metastasis in different histological types and with different depths of invasion were statistically analyzed.Results There were 46 227 lymph nodes collected from 1615 cases,with an average of 28.62 per case. Lymph node metastasis was found in 1046(64.77%) cases of gastric cancer and 10 173(22.01%) lymph nodes. Different TNM stages showed different rates of metastasis: T1(M)2.78%,T1(SM)13.61%,T2 55.59%,and T3 84.84% respectively,while the metastatic rate for histolgically well-differentiated carcinoma was 50.25%,and for low-differentiated earcinoma was 78.95%,and both had statistically significant difference(P

Objective To analyze dynamic features of TCM syndrome elements according to the neuropsychological characteristics of cognitive function impairment after ischemic stroke.Methods The hospitalized cerebral infarction patients without dementia were evaluated dynamically with the mini-mental state examination (MMSE) and Montreal Cognitive Assessment (MoCA) (Beijing Version) after IQCODE screens at different phase points.And they were divided into groups according to the severity of cognitive function impairment.The occurring frequency of TCM syndrome elements at all phase points for diagnosis was compared.Results The phase points and TCM syndrome elements closely related with the cognitive function impairment after ischemic stroke were as following:the 3rd day with fire and yin deficiency,the 7th day with yin deficiency,the 1st month with fire,phlegm,and blood stasis,and the 3rd month with fire,phlegm,blood stasis,qi deficiency,and yin deficiency.Conclusion The neuropsychological characteristics of cognitive function impairment of ischemic stroke are closely related with TCM syndrome elements and their evolvement as well as their different combinations.

Objective To evaluate the effects of hydroxyethyl starch (HES) on the renal function and blood coagulation of patients with acute pancreatitis (AP). Methods 151 patients with acute pancreatits were divided into treatment group (treated with HES, n=62) and control group (treated with crystalloid solutions, n =89). The two groups were compared by retrospective analysis of the indices such as serum creatinine (Scr), blood urea nitrogen (Bun), platelet count(Plt), prothrombin time activity (PTA), international normalized ratio (INR), activated partial thromboplastin time (APTT) and fibrinogen (FIB) before and after treatment. Results Before hospitalization, there were insignificant differences in respect of Scr and Bun in the two groups respectively. But at discharge, Scr and Bun in the treatment group were significantly increased (P<0.05), while Bun was significantly decreased 24 h right after infusion of crystalloid solutions in the control group (P<0.05). There were no differences between the two groups in terms of Scr and Bun. As for indices of coagulation, APTT in the treatment group was significantly increased after infusion of HES (P<0.05). The hospital stay in the treatment group was longer than the control group (P < 0.05). Conclusions HES may adversely affect renal function and intrinsic pathway of blood coagulation of patients with AP. Renal function and blood coagulation should be monitored when using HES.

Objective:To construct and express the fusion protein between ZZ affinity peptide and alkaline phosphatase and examine its biological activities.Methods:The alkaline phosphatase gene was cloned into pEZZ 18 vector containing ZZ peptide gene resulted in the pEZZ-AP recombinant vector.Then the vector was transformed and expressed in E.coli DH5α.And HisTrap affinity chromatography was employed to separate and purify the target protein.After analyzed by Western blot , ZZ-AP fusion protein was applied to immunocytochemistry as an alterative second antibody.Results:The result of SDS-PAGE showed that the fusion protein with a relative molecular weight was consistent with the theoretical values (62 kD).The fusion protein has rabbit IgG-binding ability and en-zymatic activity of alkaline phosphatase ,those were validated in Western blot;and it produced a good signal that was comparable in its staining pattern to that generated with goat anti-rabbit IgG-AP in immunocytochemistry.Conclusion: The ZZ-AP fusion protein was constructed successfully ,it has rabbit IgG-binding ability and enzymatic activity of alkaline phosphatase.We anticipate that the ZZ-AP fusion protein has a potential application in immunoassay field.

Background Gene encoding optineurin (OPTN) is a causative gene for glaucoma and amyotrophic lateral sclerosis,with a more expression in retina.Our previous study isolated OPTN-interacting proteins and identified that the gene encode the basic leucine zipper (bZIP) transcription factor neural retina leucine (NRL) zipper,a causative gene for retinitis pigmentosa,and further study demonstrated the interaction between OPTN and NRL proteins in nuclei of cultured HeLaS3 cells.Objective This study was to determine the protein binding site of OPTN necessary for NRL binding.Methods A deletion series of OPTN-expression plasmids were constructed and co-expressed with hemagglutinin (HA)-tagged NRL in HeLaS3 cells,respectively.The cytoplasmic and nuclear fractions were used to perform co-immunoprecipitate (CoIP) and Western blot with anti-tag antibodies.Results In the nuclear fractions of cells transfected with the del1 st,del2nd or del3rd plasmid,a band of coimmunoprecipitated HA-labelled NRL (HA-NRL) was detected.However,the del4th plasmid did not produce a band.The NRL band was not found in cytoplasmic fractions from transfected cells with any of the deletion plasmids or with the whole-length OPTN plasmid.Conclusions The protein binding site of OPTN necessary for NRL binding is determined.This result demonstrates the binding of Flag-OPTN and HA-NRL in HeLaS3 cells.A series of partial-deletion OPTN plasmids demonstrated that the tail region (423-577 amino acids) of OPTN was necessary for binding with NRL.

The therapeutic effects of intensive insulin therapy in treatment of traumatic shock combined with multiple organ dysfunction syndrome (MODS) were investigated. A total of 114 patients with traumatic shock combined with MODS were randomly divided into two groups: control group (n=56) treated with conventional therapy, and intensive insulin therapy group (n=58) treated with conventional therapy plus continuous insulin pumping to control the blood glucose level at range of 4.4-6.1 mmol/L. White blood cells (WBC) counts, prothrombin time (PT), serum creatinine (SCr), alanine aminotransferase (ALT), serum albumin and PaO(2) were measured before and at the day 1, 3, 5, 7 and 14 after treatment. The incidence of gastrointestinal dysfunction, the incidence of MODS, hospital stay and the mortality were also observed and compared. After intensive insulin therapy, the WBC counts, SCr, ALT and PT were significantly reduced (P<0.05), but the level of serum albumin was significantly increased (P<0.05) at the day 3, 5, 7 and 14. In the meantime, the PaO2 was significantly elevated at the day 3, 5 and 7 (P<0.01) after intensive insulin therapy. The incidence of gastrointestinal dysfunction, the incidence of MODS, the length of hospital stay and the mortality were markedly decreased (P<0.01). The results suggest early treatment with intensive insulin therapy is effective for traumatic shock combined with MODS and can decrease the length of hospital stay and the mortality.

Objective:To prepare the site-specific biotinylation of enhanced green fluorescence protein with double biotin molecules using Avi-tag technology.Methods:The EGFP gene was prepared by PCR and cloned into pdi-Avitag resulting the vector pEGFP-( Avitag) 2.The fusion protein EGFP-( Avitag ) 2 was expressed in E.coli DH5αand purified by employing IMAC.The site-specific biotinylation was implemented by BirA enzyme in vitro, and then was identified by competitive ELISA and Western blot.Results:The recombinant prokaryotic expression vector pEGFP-(Avitag)2 was correctly constructed,and EGFP-(Avitag)2 fusion was successfully expressed in E.coli DH5α.The results of competitive ELISA and Western blot showed that the EGFP-( Avitag) 2 could be site-specific biotinylation with double biotin molecules based on Avi-tag technology.Conclusion: The site-specific biotinylation of EGFP with double biotin molecules is successfully prepared,and we anticipate that can be used for BAS to improve the sensitivity and specificity of immunosensors.

Objective To investigate the dynamic changes of three types of anti-poliovirus neutralizing antibodies and anti-hepatitis A virus (HAV) IgG antibody in children who were immunized with inactivated enterovirus 71 (EV71) vaccine (human diploid cell).Methods Serum samples were collected from the subjects immunized with inactivated EV71 vaccine.Neutralizing antibodies against EV71 and poliovirus were detected by micro-cytopathic effect neutralization test.Enzyme linked immunosorbent assay (ELISA) was used to detect IgG antibody against HAV.Results The geometric mean titers (GMTs) of anti-EV71 neutralizing antibody increased to 4.85 following the first-dose injection of inactivated EV71 vaccine.A significant increase of GMTs (up to 64.37) could be observed 28 days after the second-dose vaccination.Meanwhile, results of the dynamic monitor showed that there were slight fluctuations in the neutralizing antibodies against three types of poliovirus on day 28 (28 days after the first-dose vaccination) compared with those on day 0 (before vaccination) (P<0.05);types Ⅰ and Ⅲ anti-poliovirus neutralizing antibodies on day 56 (28 days after the second-dose vaccination) remained slightly different from those on day 0 (P<0.05), but type Ⅱ anti-poliovirus neutralizing antibody on day 56 had restored to normal level (P>0.05).The level of anti-HAV IgG antibody was stable and no significant difference was found during the observation period (P>0.05).Conclusion This study shows that inactivated EV71 vaccine has no impact on anti-HAV IgG antibody in Children during the two-dose vaccination and in anti-EV71 antibody-producing period, but has slight influence on the anti-poliovirus antibodies.In general, changes in antibody profile do not affect the clinical efficacy of immune response.