OBJECTIVE:To study the effect of flavonoids of Astragalus complanatus(FAC) on D-galactose-induced sub-acute aging in mice. METHODS:The subacute aging mice model were established with the use of D-galactose,and the effect of FAC on the associated indexes in model mice such as enzymes,immune organ indexes,hypoxia tolerance time,swimming time under hypoxic condition etc were recorded. RESULTS:FAC treatment significantly up-regulated the activity of SOD,down-regulated MDA content,markedly increased the spleen index and thymus index,prolonged the surviving and swimming time markedly under anoxia condition. CONCLUSION:FAC has anti-aging effect and it can enhance mice's abil-ity against hypoxia and fatigue,which is probably associated to the enhancement of the nonspecific immunity in mice.

Objective To assess the effects of zinc protoporphyrin (ZnPP), an inhibitor of the heme oxygenase (HO), on the colonic interstitial cells of Cajal (ICC) of diabetic rats with colonic slow transit. Methods Diabetes mellitus (DM) model was induced by intraperitoneal injection of streptozotocin (STZ) in Sprague-Dawley rats. Twenty four successfully established DM rats were selected, and 16 healthy rats were served as controls. Six weeks later, gastrointestional (GI) dysfunction was observed by charcoal propulsion experiment in 8 DM rats and 8 controls. The rest rats in DM group were divided into 2 groups: DM rats intraperitoneal injected with PBS (n=8) or with 10 μmol/kg of ZnPP (n = 8) every other day for 3 weeks. The rats in control group (n = 8) were intraperitoneally injected with PBS. The levels of HO and c-kit (the special receptor of ICC) expression were detected by Western blotting. The distribution of ICC was observed by immunohistochemistry and the area of c-kit positive cells was counted. Results The GI propulsion rate in DM rats interfered with PBS was significantly declined compared to that in the controls (63.0%± 1.2% vs 71.8%±2.0%, P<0.05). But it was improved in DM rats interfered with ZnPP (72.5± 2.6%, P<0.05), which showed no significant differentee with that in control group (P>0.05). The expression of HO-1 in close and distant colon of DM rats interfered with ZnPP was decreased (P< 0.05). The expression of HO-2 in close colon and the area of c-kit positive cells of DM rats interfered with PBS was reduced compared with that in controls (P<0.05), but both were improved in DM rats interfered with ZnPP (P<0. 05). Conclusion Administration of ZnPP might be able to protect ICC by its blockage of HO-1 in DM rats with gastrointestinal dysfunction.

Objective To analyze and compare the fluorescence in situ hybridization(FISH) and immunohistochemical(IHC) for detecting HER-2 gene amplification and protein expression in breast cancer tissues .Methods 110 cases of breast cancer from Janu-ary 2008 to May 2012 receiving the modified radical mastectomy were selected .The resected breast cancer tissue was detected by FISH and IHC and the detected results were performed the comparative analysis .Results Among 110 cases of breast cancer tissue , 25 cases(22 .73% ) were the HER-2 protein expression(+ + + ) ,44 cases(40 .00% ) were(+ + ) ,26 cases(23 .64% ) were(+ ) and 15 cases(13 .64% ) were(-) .Among 110 cases ,the gene amplification was in 28 cases(25 .45% ) and no gene amplification was in 82 cases(74 .55% ) .The positive(+ + + ) of the IHC detection was coincident with that of FISH ,and the negative(+ /-) of the IHC detection was also coincident with that of FISH ,there was statistical difference between the suspicious positive of the IHC de-tection and the results of FISH (P<0 .05) .But the total coincidence of the IHC detection results and FISH test results was 89 .29%(25/28) ,and the two detection methods had the positive correlation (χ2 =84 .89 ,P<0 .01) .Conclusion The positive and negative expression of the IHC detection has better consistency with that of the FISH detection .However ,the coincidence of the IHC suspi-cious positive expression and the FISH results is poor ,indicating that the suspicious positive sample of the IHC detection needs to be detected by the FISH detection .

Objective To investigate the influence of fast track surgery(FTS) on the concentrations of serum IL‐6 ,IL‐10 , and TNF‐αin pancreatic cancer patients ,and its clinical significance .Methods Eighty patients undergoing pancreatic cancer opera‐tion were divided into FTS nursing group and traditional nursing group .The concentrations of serum IL‐6 ,IL‐10 ,and TNF‐αin dif‐ferent times before and after operation ,and the same period between groups were measured .Results (1)The concentrations of ser‐um IL‐6 ,IL‐10 ,and TNF‐αwere no significantly difference in two groups before operation(P>0 .05);the concentrations of serum IL‐6 in the first and third postoperative day were higher than those before operation in the observation group (P<0 .05);there was no significant difference of the concentrations of serum IL‐6 between the fifth postoperative day and before operation(P>0 .05);the concentrations of serum IL‐6 after operation were higher than those before operation in the control group(P<0 .05);the concentra‐tions of serum IL‐6 in the same time after operation in the observation group were lower than those in the control group(P<0 .05);(2)The concentrations of serum IL‐10 in the first ,third and fifth postoperative day were higher than those before operation in the observation group(P<0 .05);the concentrations of serum IL‐10 in the first and third postoperative day were higher than those be‐fore operation in the control group(P<0 .05);the concentrations of serum IL‐10 in the same time after operation in the observation group were higher than those in the control group(P<0 .05);(3)The concentrations of serum TNF‐αin the first postoperative day were higher than those before operation in the observation group(P<0 .05);there was no significant difference of the concentra‐tions of serum TNF‐αbetween the third and fifth postoperative day and before operation(P>0 .05);the concentrations of serum TNF‐αin the first and third postoperative day were higher than those before operation in the control group(P<0 .05);there was no significant difference of the concentrations of serum TNF‐αbetween the fifth postoperative day and before operation in the control group(P>0 .05);the concentrations of serum TNF‐α in the first and third postoperative day in the observation were lower than those in the control group(P<0 .05) .Conclusion FTS could significantly reduce inflammatory reaction ,improve immunosuppres‐sion and helps to recover .

Objective To investigate cortactin expression in malignant colorectal tissues and corresponding adjacent non-cancerous colon tissues,precancerous lesions (adenomatous polyps) and the relationship between the expression of cortactin and cell division in colorectal cancer cells.Methods The expression of cortactin was detected by immunohistochemistry in colorectal cancer,colorectal adenomatous polyp (precancerous lesions) and colorectal tissues adjacent to adenocarcinomas (normal tissues).Kaplan-Meier method was employed to compare the survival between the groups.Cortactin expression and cell division were detected by Western blot and immunofluorescence in SW-620 colon cancer cells treated with cortactin siRNA.Results The positive expression rate of cortactin was significantly higher in colorectal cancer tissues than in adenomatous polyp tissues and pericarcinomatous normal tissues.Overexpression of cortactin in colorectal cancer tissues was correlated with poor differentiation (P ＜ 0.01),lymph node metastasis (P =0.006),and TNM stage (P =0.022).The 5 year survival rate of the group of negative/weak positive expression of eortactin was significantly higher than the group of strong positive expression of cortactin.CTTN gene amplification in colorectal cancer tissues was obvious.Cortactin siRNA induction caused a lower cortactin protein expression in colorectal cancer cells.Conclusions It is suggested that the excessive expression of cortactin contributes to the growth of cancer cells in colorectal cancer.

Objective　To investigate the surgical management of a gun shot wound of blood vessels and immersied, and evaluate its primary effect. Methods　100 rabbits were divided randomly into simple wounded group(SWG,n=50) and seawater immersion group(SIG,n=50).F emoral arteries were impacted by 0.38 gram steel spheres with velocity of 600 ～800 meters per second fired by 7.62 mm rifle. Animals in SIG were immersed in artificial seawater (pH 8.2～8.4, salinity 25.4,temperature 21℃) for 60 min, o f which those in SWG were spared. Grossly injuried artery was excised and restor ation of blood flow was reconstructed by end-to-end anastomosis or reversed au togenous venous grafting or cryopreserved arterial allografting. At 24 h,7,1 4,21 days after operation, blood flow was examined by Doppler ultrasonic detecti on and part of anastomotic sites and graft were collected for pathological obser vation. Results　In completely transected injury, the patency in SIG was 80.00%,while that in SWG was 86.67% in the first 3 weeks. In arterial c ontused injury ,patency in SIG was 86.67%,and that in SWG was 82.35% at the same time. Thrombosis occurred mostly in the first postoperative week. Atypical endo thelial cells were found at the anastomosis sites in the first postoperative week, and the anastomosis sites were lined with endothelium in 3 weeks postopera tively. Conclusion　Early curative effect could be obtained. Whe n grossly injuried artery is excised and followed by a routine surgical procedur e in the treating gunshot wounds immersed in seawater.

Objective To observe the influence of the clinical pathway management intervention on medical service behaviors.Methods The clinical cases data in a grade A class 3 hospital of Zhangjiakou City,Hebei Province during 2011-2013 were retrospectively analyzed.The influence of implementing the clinical pathway management on the diagnosis results,medical records writing quality,clinical medication,operation,hospitalization time,medical costs,assisted examination items selection were observed and analyzed.Results The admission and discharge diagnosis coincidence rate and preoperative and postoperative diagnostic coincidence rate of 3 diseases entities in the two groups were 100%,the difference had no statistical significance (P>0.05);the medical record grade-A rate in the implementing clinical pathway group was significantly higher than that in the non-implementing clinical pathway group,the medical record grade-B rate was significantly lower than that in the non-implementing clinical pathway group,the difference was statistically significant (P<0.05);the antibacterial drugs cost and drugs proportion in the implementing clinical pathway group were significantly lower than those in the non-implementing clinical pathway group,the medication proportion within the directory in the implementing clinical pathway group was significantly higher than that in the non-implementing clinical pathway group,the difference was statistically significant (P<0.05);the preoperative preparation time in the implementating clinical pathway group was less than that in the non-implementing clinical pathway group,the difference was statistically significant(P<0.05),the grade-A healing rate had no significant difference between the two groups (P>0.05);the average hospitalization time,total costs,drug costs,laboratory fee and examination fee in the implementing clinical pathway group were lower than those in the non-implementing clinical pathway group,the difference was statistically significant (P<0.05);the operation fee,healthy material fee and nursing fee had no statistically significant difference was statistically significant(P<0.05).Conclusion Implementing the clinical pathway can standardize the doctor′s diagnosis and treatment behaviors and makes the medical work to develop to be more favorable for patients.

Objective To establish the A549 cell line with stable expression of HIF1α by using lentiviral vector system.Methods Primers were designed and synthesized with human HIF1α gene coding sequence by the National Center of Biotechnogical Information(NCBI) as the template.HIF1α was amplified by PCR.The HIF1α fragment recycled by enzyme digestion was recombined with prepared lentiviral vector HBLV-RFP-Puro.The recombinant plasmid was identified by PCR and gene sequencing.The recombinant plasmid and the auxiliary plasmid were co-transfect into 293T cell.After filtration and concentration of packaged virus,the viral titer was detected by using the dilution counting method.The prepared lentivirus was infected A549 cells.The drug screening was adopted to stabilize the transfected cell line.The transfection effect was detected and observed by fluorescence microscope and Western blotting.Results The HIF1α fragment amplified by PCR was successfully verified and the recombinant plasmid was successfully constructed by PCR and gene sequencing identification.High-titer LV-HIF1α was obtained by successful package.After LV-HIF1α infecting A549 cells,the cells showed the red fluorescence by fluorescence microscope.The expression level of HIF1α in the LV-HIF1α group was significant higher than that in the control group by Western blot.Conclusion The 549 cell line with HIF1α stable expression mediated by lentivirus is constructed successfully.

Objective To investigate the relationship between serum resistin,leptin and adiponectin with microangiopathy in the patients with type-2 diabetes mellitus(T2DM).Methods One hundred and twenty patients with T2DM in our hospital were selected and divided into the non-microangiopathy group (NON-MAP,60 cases) and microangiopathy group (MAP,60 cases) according to whether complicating microangiopathy.Other 60 individuals undergoing healthy physical examination were selected as the normal control group(NC).Fasting serum resistin,adiponectin and leptin levels were detected in each group.Fasting blood glucose,insulin and blood lipid levels were also detected.The insulin resistance was evaluated by using insulin resistance index(HOMA-IR).Results The levels of serum resistin,leptin,free fat acid(FFA) and hs-CRP in the NON-MVP group and MVP group were signifi cantly higher than those in the NC group,while the adiponectin level was sigrnificantly lower than that in the NC group,the difference was statistically significant(P＜0.05).The correlation analysis showed that serum resistin and leptin levels had positive correlation with hs-CRP,FFA,HOMA-IR and TG(P＜0.05),and had negative correlation with HDL-C(P＜0.05);the adiponectin level was negatively correlated with hs-CRP,FFA,HOMA-IR and TG,while positively correlated with HDL-C(P＜0.05);serum resistin and leptin levels had positive correlation,both had significantly negative correlation with adiponectin;with serum resistin,leptin and adiponectin as the dependent variables,the multiple stepwise linear regression analysis showed that HOMA-IR and waist to hip ratio had maximal influence on them.Conclusion Serum resistin and leptin levels increase and adiponectin level decrease in T2DM patients suggests that serum resistin,leptin and adiponectin are correlated with T2DM occurrence as well as microangiopathy occurrence.

Objective With the same times of injection to compare low-dose intradermal regimen with routine-dose intramuscular inoculation in revaccination of non-responders to hepatitis B vaccine. Methods 40 healthy non-responder children collected by screening were administrated a three-dose revaccination randomly by intramuscular or intradermal route (10 vs 2 g per dose), and regularly tested for serologic markers up to five years. By the end of follow-up, a booster dose (5 μg) was given to those who had lost anti-HBs of ≥10 mIU/mL (seroprotection) and anamnestic response was estimated thereafter. Results All 17 intramuscular and 22 of 23 intradermal children effected seroprotection after revaccination. Intradermal children lost seroprotection over time significantly rapider compared with intramuscular children (Log Rank test, P= 0.029). In year 5, 50% of intramuscular but only 18.2% of intradermal children still maintained seroprotection (P=0.075). 12-14 days after the booster dose, all the eight intramuscular children developed an anamnestic response with anti-HBs titer increasing greater, but two of the 18 intradermal children failed to mount seroprotective level. Conclusion Three-routine-dose intramuscular revaccination was significantly effective than low-dose intradermal one with the same times of injection, especially in long-term immunity. We recommend routine-dose intramuscular protocol in revaccination of non-responders.