Objective To prepare the composite of Polycaprolactone(PCL) and Bone powder(BP) for bone repairing,and evaluate the biocompatibility of the material.Methods The materials were prepared in college of chemical engineering and materials science,Shanghai Jiaotong University,and the animal tests were fulfilled in Animal Experimental Center of the Second Military Medical University of PLA from June 2010 to March 2011.The PCL and BP were blended together to get the copolymer by haake ( Rheocord 900 ) torque rheometer.Then the extracts of the material in DMEM for 24 hours were obtained.Firstly biological safety detection of the materials was performed:①cytotoxicity test:the extracts were mixed with DMEM to seed rabbit osteoblasts for 24 hours.The cell proliferation was detected by MTT method.The cell proliferation rate and toxicity grate were also calculated ( cytotoxicity of grade 0 or 1 was qualified).②hemolysis test:the extracts were mixed with fresh rabbit blood,hemolysis rate were calculated ( hemolysis mte less than 5％ was qualified).③pyrogenic test:the extracts were injected into the rabbit ear vein with the dose of 10 ml/kg.The anus temperature was measured per hour for three times.The elevator degree was calculated by subtracting the natural temperature from the highest one.Secondly,cellular adhesion test of the material composite was performed.The composite was casted on the cover glasses.The samples were put in 6-well tissue culture plates,totally 6 × 104 osteoblasts were seeded each well and cultured for 3 days.The cells were observed at 24 and 72 hours of culture,and photography was performed using inverted phase contrast microscope.④general toxioity test:the extracts were injected into mouse vein with the dose of 50 ml/kg.The toxic symptoms were observed within 72 hours.Results The novel copolymer of PCL/BP was prepared.①Biological safety:the cell viability of the composite was 87.8％ and cytotoxicity was grade 1.The hemolysis rate was 2.4％,less than 5％.The maximal variety of body temperature in rabbits were 0.4℃,which accorded with the national standard ( ＜0.6℃ ).The general toxicity test showed that there were no adverse effects in the animals,such as death,convulsion,paralysis,respiratory depression,diarrhea and decreasing weight.②Cell adhesion:adhesive cells manifold obviously and assembled 24 hours later,inosculated gradually 3 days later,and the cells were in good fettle.Conclusion The composite of PCL/BP accords with cytotoxicity demand of the biological materials,presenting no toxicity,pyrogenicity or hemolysis.It possesses good biocompatibility and cellular adhesion.

OBJECTIVE:to provide reference for the continuous improvement of surgical site infection (SSI). METHODS:There were totally 7 472 patients with typeⅠand typeⅡincision surgeries in a hospital after the targeted monitoring and special recti-fication(Jul. 2012 to Jun. 2013,monitoring group)and 5 958 patients with surgeries during the same period before special rectifi-cation(Jul. 2010 to Jun. 2011,control group). The clinic data of typeⅠand typeⅡincision surgical was compared,including infec-tion,the perioperative antibiotics use and hospitalization time after surgery,etc. RESULTS:The infection rate of typeⅠand typeⅡstandardized incision in monitoring group was respectively 0.35% and 0.43% and control group was respectively 0.60% and 1.36%(P<0.05). The rate of typeⅠincision perioperative antibiotics use in monitoring group was 10.72% and control group was 86.88%(P<0.05). The post-operative non-medication rate of patients was increased from the 6.98%(control group)to 49.20%(monitor-ing group)(P<0.05),the discontinuance rate within 48 h was increased from the 32.09%(control group)to 44.11%(monitoring group),and the ratio of patients who took antibiotics after the surgery for more than 3 d was decreased from the 42.82%(control group)to 3.05%(monitoring group)(P<0.05). The inguinal hernia repair time in monitoring group was 3.90 d,shorter than con-trol group(4.22 d)(P=0.018). The patient with gallbladder surgery in monitoring group was 6.47 d,compared with control group (6.38 d),there was no significant difference (P=0.619). CONCLUSIONS:The special rectification can obviously promote the standardized of perioperative antibiotics use,reduce the incidence of SSI and shorten the hospitalization time after operation.

Objective To investigate the effects of keratinocyte growth factor on peritoneal adhesion formation in rats. Methods Thirty Sprague-Dawley( SD) female rats were randomly assigned to 3 groups, KGF group( n = 10), positive control group (n = 10) , and negative control group (n = 10). Seven days after surgery, rats were killed and the adhesion degree was evaluated by Leach scale. Immunohistochemical technique was used to identify the expression of tPA and PAI-1. Stained with HE,the histomorphology changes of the adhesion tissue were observed by light microscope. Picrosirius-polarization method was used to observe the expression of type Ⅰ or Ⅲ collagens in two groups. Results In the KGF group,lower collagen fibers were noted and the gross adhesion scores was significantly lower than that in positive control group (4. 8 ± 1. 0 vs 7. 6 ± 1. 0; t = 5.422; P ＜ 0. 01). The expression level of type Ⅰ collagens was significantly lower in the KGF group than in positive control group (69 ±11 versus 55 ±9;t = 3. 214 ;P ＜0. 01) ,but there was no significant difference in the expression of type Ⅲ collagens among the two groups (48 ± 7 versus 50 ± 10; t = 0. 481; P ＞ 0. 05). The immunohistochemistry showed that the expression of tPA significantly increased in the KGF group than in positive control group and negative control group(88 ±4.0 versus 112 ±4.0, 101 ±2.0;F = 109. l,P＜0. 01) , However, no statistically significant difference for the expression of PAI-1 was noted among the three groups ( F = 1. 391, P ＞ 0. 05). Conclusions Keratinocyte growth factor promotes mesothelium repair, increases mesothelial fibrinolytic activity, inhibits the deposition of collagen and reduces the intensity of postoperative adhesions.

Objective To study the corelation between the expression of COX-2 and postoperative adhesions and to determine the effect of COX-2 selective inhibitor,Celecoxib,on postoperative adhesion formation. Methods Fifty SD rats were randomly assigned to five groups each consisting of 10 rats,Study groups were as follows:(A)positive control group,(B)sodiumhyahronate group,(C)low dose Celecoxib group,(D)high dose Celecexib group,(E)negative control group,Five rats in each group were treated accordingly for consecutive 8 days and 15 days respectively and sacrificed,After treatment,intra-abdminal adhesions were scored using a standard method.The adhesions tissure and injured peritonaeum were subjected to Westem-blotting to detect the expression of COX-2. Results The level of postoperative adhesions and expression of COX-2 of sodiumhyahronate group、low dose Celecoxib group、high dose Celecoxib group were lower than that of positive control group(P<0.05),Sediumhyahronate treatment group was different formthe two Celecoxib treated groups(P<0.05).Conclusions Selective COX-2COX-2.mechanism,provides durable inhibition of intra-abdominal adhesions through an antiangiongenic COX-2 mechanism.

Objective:To illuminate the influence of InsB15-23 H-2Kd dtSCT to the morbidity of type 1 diabetes mellitus in NOD mice.Methods:An eukaryotic plasmid encoded membrane-expressed InsB15-23 H-2Kd dtSCT was inoculated into 3 weeks old female NOD mice subcutaneously and the blood sugar and morbidity of type 1 diabetes mellitus were monitored once a week.To illuminate the cellular mechanism of immunologic intervention of membrane-expressed InsB15-23 H-2Kd dtSCT to the course of type 1 diabetes mellitus in NOD mice,the mononuclear cell infiltration of islets was detected by tissue slice and the frequency of IGRP206 2-14 specific CTLs in PBMC was analyzed by FACs.Results: As compared with pcDNA3.1 (-) control ( 60%) and untreated NOD mice ( 80%) , mice immunized with InsB15-23 H-2Kd dtSCT exhibited low level of islet infiltration and low morbidity in 30 weeks old ( 9%) .But the frequency of IGRP206-214 specific CTLs in PBMC of 16 and 40 weeks old mice showed no difference.Conclusion:Membrane-expressed InsB15-23 H-2Kd dtSCT can protect NOD mice from type 1 diabetes mellitus in IGRP206-214 independent pattern.

Objective To investigate the eftect of radiation dose of dual-source computed tomography (CT) dual energy single-phase enhanced scan in patients with esophageal cancer.Methods The prospective study was conducted.The clinicopathological data of 56 patients with esophageal cancer who were admitted to the Lishui Hospital of Zhejiang University between January 2015 and December 2016 were collected.All the patients were divided into the experimental group (undergoing dual-source CT dual energy single-phase enhanced scan) and control group (undergoing dual-phase CT enhanced scan) bv randomised block method.TNM classification of esophageal cancer (Seventh Edition) published by American Joint Committee on Cancer (AJCC) was used as a standard TNM staging.Two observers independently read films.All the patients underwent radical resection of esophageal cancer or palliative surgery,and then received adjuvant radiochemotherapy.Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival of patients up to March 2017.Observation indicators:(1) consistencies of T staging,N staging and M staging;(2) accuracies of T staging,N staging and M staging (pathological results as a gold standard);(3) radiation dose of CT scan;(4) treatment and follow-up situations.The Kappa test was used for evaluating the consistency,κ≥0.75 as a good consistency,0.40≤κ＜0.75 as a normal consistency and κ＜0.40 as a poor consistency.Comparisons of count data and ratio were done by the chi-square test.Comparisons of measurement data were analyzed by the t test.Results A total of 50 patients were enrolled in the study,including 25 in the experimental group and 25 in the control group.(1) Consistencies of T staging,N staging and M staging:all the 50 patients finished successfully CT scans.Two observers considered that consistencies of T staging,N staging and M staging in the 2 groups were normal (κ =0.452,0.618,0.729,P＜0.05).Consistencies of N staging and M staging were superior to T staging.(2) The pathological results were used as a gold standard.Accuracies of T staging,N staging and M staging in the experimental and control groups were 72％,76％ and 88％,84％ and 92％,88％,respectively,with no statistically significant difference between the 2 groups (x2 =0.10,0.37,0.50,P＞0.05).(3) Radiation dose of CT scan:volume CT dose index (CTDIvol),dose length production (DLP) and effective radiation dose (E) were (10.35±2.01) mGy,(400.63± 34.13) mGy · cm,(5.61 ± 0.47) mSv in the experimental group and (3.55 ± 0.60)mGy,(140.66± 10.89) mGy · cm,(1.98±0.17) mSv in the control group,respectively.There were statistically significant differences in CTDIvol and E between the 2 groups (t =16.23,36.30,P＜0.05).(4) Treatment and follow-up situations:of 50 patients,43 patients received treatments,including 32 undergoing radical resection (11 receiving postoperative adjuvant chemotherapy),6 undergoing palliative surgery,3 receiving single radiotherapy and 2 receiving single chemotherapy.Thirty-six of 43 patients were followed up for 3-18 months,with a median time of 6 months.During follow-up,1-year survival rate was 61.1％.Conclusion Dual-source CT dual energy single-phase enhanced scan in patients with esophageal cancer cannot reduce accuracy of TNM staging,but decreased effectively radiation dose.

Objective To clarify the efficacy and safety of cyclosporine A CsA) as salvage therapy in patients with severe active ulcerative colitis (UC) and refractory to steroids.Methods A total of 24 severe active UC patients refractory to steroids and hospitalized from 2006 to 2012,were retrospectively enrolled.Data including demographic features,clinical manifestations,laboratory tests and medications were collected.Results CsA was effective in 15 (62.5％) patients,who did not receive colectomy during 12-week administration.This regimen was tolerable in most patients.Twelve (50.0％) patients reported 16 adverse events,but only one patient withdrew CsA due to intolerance.The rates of adverse events in initial intravenous CsA including 4 mg · kg-1 · d-1,3 mg · kg-1 · d-1 and 2 mg· kg-1 · d-1 were 2/2,9/17 and 1/5 respectively.Responders had higher white blood cell count compared with non-responders (P =0.045).Conclusions CsA could be an effective alternative regimen to colectomy in severe active UC patients who are refractory to steroids.

ABSTRACT:Objective To assess the effect of prolonged laparoscopic surgery on peritoneal mesothelial cells and fibrinolysis in humans.Methods We examined prospectively 1 6 consecutive patients who underwent laparoscopic surgery (LAP)and 2 1 patients who underwent conventional open surgery (OP)for high-medium rectal cancer with curative intent.During the procedure,biopsy of the parietal peritoneum was made before operation and at 45 min,90 min,and 120 min after operation.The tissue-type plasminogen activator (tPA)and plasminogen activator inhibitor-1 (PAI-1 )were determined by enzyme-linked immunosorbent assay in peritoneal tissues.The cellular injury was detected by LDH assay.The proliferation was quantified by MTT assay.Results PAI-1 activity in the peritoneal tissue was significantly lower in LAP group than in the OP group.tPA activity decreased after 45min of open surgery,but there was no significant change in the LAP group.With time extension,the LDH activity increased and the proliferation of the mesothelial cells decreased.Conclusion Preservation of a prolonged hypofibrinolytic state by inhibition of PAI-1 up-regulation during LAP may predispose patients to less postoperative peritoneal adhesion. The cellular injury becomes apparent and the proliferation is inhibited during prolonged laparoscopic surgery.

This study was aimed to construct Sirt1 shRNA interfering vector and to analyze the effects of Sirtl on cell proliferation and apoptosis in HepG2, A549 and 293T cell lines. To design and synthisize Sirtl shRNA sequence then recombinate it to pGenesil-1.0 plasmid, the positive pGenesil-1.0-Sirtl vector clone was screened by effective detections and sequencing. The vectors were transfected into HepG2, A549, 293T cell lines, and Sirtl expression levels in these clones were detected by RT-PCR and Western-blot. These clone cell proliferation activities were detected by MTT, and these cells apoptosis incidences were detected by MTT experiment after treated with DOX. The results showed that Sirt1 shRNA interfering vectors were successfully screened. The levels of Sirtl expression in HepG2-sh, A549-sh and 293T-sh cells were significantly reduced compared with their control cells. It was indicated that the proliferation activities of these cells were impaired and anti-apoptosis capabilities of HepG2-sh, A549-sh and 293T-sh were also impaired notably. Sirt1 took an important role in maintaining cell proliferation and resisting cell apoptosis caused by DNA damage, and this result also provided theoretical information for the further research.
Apoptosis ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; Sirtuin 1 ; genetics ; Transfection

Objective To evaluate the feasibility of needle-based confocal laser endomicroscopy (nCLE) for imaging of intra-abdominal tissues and organs in rabbit models in vivo. Methods The nCLE miniprobe was inserted through the 19-gauge needle into various intra-abdominal tissues and organs[omentum majus, liver, pancreas and psoas major (skeletal muscle)]. The nCLE images were acquired and real-time sequences of respective locations were recorded. Finally, nCLE image characteristics were compared with histopathologic findings. Results nCLE was successfully performed in intra-abdominal tissues and organs of five rabbit models. The microscopic structures of cells, glands and microvessels in the omentum majus, liver, pancreas and psoas major ( skeletal muscle) were visualized clearly, respectively. Characteristics of various intra-abdominal tissues and organs were displayed on nCLE images, which were correlated well with histological findings. Conclusion Imaging of intra-abdominal tissues and organs with nCLE in vivo is feasible in future clinical practice.