Objective To investigate the repairative effect of local high expression of glial cell line-derived neurotrophic factor (GDNF) in the repair of sciatic nerve defects. Green fluorescent protein (GFP) gene was used as reporter gene. Two groups including gel matrix control group and gel matrix plus MSCs group were set ( 10 rabbits per group). Methods GDNF secreting mesenchymal stem cells (MSCs) mediated by lentiviral vectors were suspended in Matrigel to produce the artificial graft, and then it was locally implanted to a 20 mm-long sciatic nerve defect in rabbits. Results In the GDNF group,the nerve continuity was restored in seven rabbits and the functional amelioration of the wounded limb was apparent. While there found no restoration of the nerve continuity in the control groups. The migration distance of GDNF secreting MSCs was 20 mm, suggesting a good integration of the transplanted cells with host cells. The glial fibrillary acidic protein (GFAP) + green fluorescent protein (GFP) and -100 protein GFP accounted for 60% and 70% respectively in the experimental group, while it was only 40% and 30% in gel matrix plus MSCs group. Conclusions Local implantation of GDNF secreting MSCs can promote the regeneration and functional recovery of the sciatic nerves. GDNF seems to play an important role in inducing transdifferentiation of MSCs to Schwann cell lineage.

Objective To investigate the effects of laparoscopic tube sparing operation in the treatment of tubal ectopic pregnancy. Methods Clinical data of 56 cases of tubal pregnancy undergoing laparoscopic conservative treatment were retrospectively reviewed. Results Salpingectomy was required in 8 cases because the electrocoagulation had proven ineffective for the bleeding of operated area, whereas the tube sparing surgery was accomplished successfully in the remaining 48 cases, in 3 of which one treatment course of MTX was administered on account of recurrent episodes. Hysterosalpingography with iodized oil at 1～2 postoperative months in 47 cases receiving conservative surgical treatment revealed that, the involved oviduct was patent in 36 cases, unobstructed but not patent in 8 cases, and obstructed in 3 cases, while the contralateral oviduct (which had been previously resected in 3 cases) was patent in 34 cases, unobstructed but not patent in 7, and obstructed in 3. Conclusions Under proper conditions, laparoscopic tube sparing operation is effective in the treatment of tubal ectopic pregnancy.

Objective To study the effects of Xuanbaichengqi decoction on the regulation of inflammatory factors TNF-α and IL-1 β in patients with acute respiratory distress syndrome (ARDS) and its mechanism.Methods 60 patients with ARDS who had been hospitalized from January 2016 to August 2016 were randonly selected and then assigned to a control group (30 patients) and a study group (30 patients).Both groups received routine therapies and mechanical ventilation,and the study group received additional Xuanbaichengqi decoction.Levels of TNF-o,IL-1β and IL-10,and peak inspiratory pressure (PIP),plateau airway pressure (Pplat),PVC,forced vital capacity (FVC),forced expiratory volume in one second (FEV 1),and oxygenation index (PaO2/FiO2) were detected before and after treatment.Results Levels of TNF-α and IL-1βwere significantly lower in the study group than in the control group (P ＜ 0.05).IL-10 level was significantly higher in the study group than in the control group (P ＜ 0.05).And the improvement in PIP and Pplat was also significant in the study group (P ＜ 0.05).Less adverse reactions occurred in the study group.Conclusions Xuanbaichengqi decoction can improve the indexes for respiratory mechanics and pulmonary function in patients on mechanical ventilation and reduce the inflammatory response,having a significant effect.

OBJECTIVE: To observe the effect of perioperative administration of rhubarb on the acute inflammatory response in patients with gastric cancer. METHODS: In this prospective, single-blinded, controlled clinical trial, thirty-one patients with gastric cancer operatively treated were randomly divided into two groups, with 14 patients in control group and 17 in study group. Patients in both groups were given an isocaloric and isonitrogenous enteral diet. The enteral diet was started 36 hours after operation, and continued for 6 days. Patients in the study group were fed with rhubarb before operation, and at 1 day and 2 days after operation. Indexes of acute inflammatory response such as serum C-reactive protein (CRP), interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha), and indexes of nutritional status such as serum albumin (ALB), prealbumin (PA) and transferrin (TRF) were measured before operation, and at 1 day, 3 and 7 days after operation. RESULTS: Patients in both groups had acute inflammatory response, and the indexes of nutritional status decreased after operation.IL-6, CRP and TNF-alpha tested at 3 and 7 days after operation were lower in the study group as compared with those in the control group, and the recovery time of gastrointestinal motility such as borborygmus, gas elimination and defecation was shorter in the study group as compared with that in the control group. The indexes of nutritional status showed no significant differences between two groups after operation. CONCLUSION: Rhubarb can positively modulate the acute inflammatory response, promote the recovery of postoperative gastrointestinal motility, and benefit enteral nutrition support in patients who have undergone major operations for gastric cancer.

The combined teaching method of case-based learning and W2 H2 thinking-type learning was used in the comparative morphology experiment teaching.The teaching method can further strengthen the reform of the compar-ative morphology experiment teaching, and improve the quality of practice teaching.

OBJECTIVE:To establish a method for the simultaneous determination of 7 active constituents in Tangshen qing-du granule. METHODS:HPLC was performed on the column of SHIMADZU Inert Sustain C18 with mobile phase of acetonitrile-0.1%phosphoric acid at a flow rate of 1.0 mL/min,detection wavelength was 327 nm for chlorogenic acid and caffeic acid,280 nm for baicalin,228 nm for arctiin and 276 nm for wogonoside,baicalein and wogonin,column temperature was 35℃,and injection volume was 10 μL. RESULTS:The linear range was 4.830-154.6 μg/mL for chlorogenic acid and(r=0.9998),0.750-24.1 μg/mL for caffeic acid(r=0.9997),22.859-731.5 μg/mL for baicalin(r=0.9997),8.491-271.7 μg/mL for arctiin(r=0.9993),2.471-79.0μg/mL for wogonoside(r=0.9996),6.656-213.0 μg/mL for baicalein(r=0.9994) and 2.756-88.2 μg/mL for wogonin (r=0.9998);RSDs of precision,stability and reproducibility tests were lower than 2.0%,recoveries were 96.86%-100.82%(RSD=1.46%,n=6),98.79%-101.09%(RSD=0.93%,n=6),97.57%-101.51%(RSD=1.37%,n=6),97.76%-99.63%(RSD=0.77%,n=6),97.99%-100.12%(RSD=0.76%,n=6),96.54%-101.07%(RSD=1.87%,n=6) and 96.60%-99.59%(RSD=1.14%,n=6). CONCLUSIONS:The method is simple with good precision,stability and reproducibilty,and can be used for the simultaneous determination of 7 active constituents in Tangshen qingdu granule.

Objective:To construct the yeast expressive vector of rmFⅦ, in which mFⅦ was mutated to inhibit coagulation without affecting the affinity for TF, and express it in Pichia pastoris. Methods:The full length cDNA encoding mFⅦ was amplified from a mouse liver by RT-PCR method, site-direct mutated and restriction enzyme digested as design. Cloning into pPIC9K, electroporation of Gs115, in vivo screen of multiple inserts by G418 resistance, BMGY/BMMY are used for induction and expression of rmFⅦ in pichia pastoris. These proteins were also screened for functional activity. Results: Three different rmFⅦ-pPIC9K yeast expression vectors and it's aim protein were obtained,two kinds of proteins were found to be functional active as design. Conclusion:rmFⅦ protein can be expressed in pichia pastoris and it might facilitate the development of tumor-target molecule, and novel anti-agiogenesis drug study.

Objective To construct and identify the recombinant vector pcDNA3. 1 (-) B/myc-BRMS 1 carrying breast-cancer metastasis suppressor 1 (BRMS 1) which can express in eukaryote cells and which will provide the basis for further researching the mechanisms of metastasis suppression and working on cancer metastasis gene ther-apy. Methods To isolate total RNA from MCF - 7 cells and design a pair of primers, and coding sequence of aRMS 1 cDNA were amplified from human breast cancer cells MCF -7 by reverse transcription-polymerase chain reaction (RT-PCR). Then the product was inserted to the PcDNA3. 1/myc-His (-) B plasmid. The recombined pcDNA3. 1 (-)B/myc-BRMS1 was identified by gene sequence analysis,then recombinants was transfected into HEK-293 cells and was identified by Western blot. Results The recombinant of pcDNA3.1 (-) B/myc-BRMS1 was structurally confirmed by analysis of sequencing. The inserted fragment in the vector was in the right direction and its sequence was structurally confirmed to be consistent with CDS sequence of human BRMSI cDNA that of the published data. GenBank, [AF159141]. The recombinants was transfected into HEK-293 cells ,then the cells expressed protein tagged c-myc identified by Western blot indicated it can express in eukaryote cells. Conclusion cDNA of human BRMS1 can be successfully cloned and inserted into Eukaryote-expression vector. The newly constructed vector may serve as the potential tool to conduct further comprehensive experiments in future on BRMS1 function and on gene therapy.

Objective To study the role of total flavonoids of hippophae rhamnoides in improving contactile function of stretched cardiac myocyte.Method Flavonoids were given to stretched myocytes which were proved their contractile function decline and then myocyte contractile mechanics characteristics and calcium transfer were measured.Result Flavonoids increased myocyte contractility,as indicated by myocyte shortening,velocity of shortening,peak+dL/dt and peak-dL/dt during shortening,in a concentration-dependent manner (r>0.9,P<0.001),and with no relation to the intracellular calcium transfer in the myocytes.Conclusion Flavonoids of the traditional Chinese drug hippophae rhamnoids is effective in improving the contractile function of stretched cardiac myocyte in low dosage.

Covered in the paper are the policies and progress of the county-level public hospital reform in the province,and a summarized analysis of the self-appraisal reports and hospital reform statements submitted by the health authorities and up to 300 hospitals in 79 counties of the province.As found in the papers,the ongoing reform in Zhejiang is focused on reforming the business models in place,seeking breakthroughs from the zero mark-up policy on drug sales,in addition to such policies as reducing drug costs,adjusting medical service pricing,financial subsidies,and medical insurance payment reforms.Despite the initial success,further policy studies are needed in terms of internal management,upper and lower linkage,and personnel incentives.