Objective To purify BMSCs by a simple Colony-Forming system and identify BMSCs in vitro.Methods BMSCs were planted at low-density ( 10/cm2 ) in the early stage of isolation,additional cells were scraped until a colony formed,secondly seeded those Colony-Forming cells at low-density.This process was repeat again when passaged cells formed another colony.This so called Colony-Forming selection was repeated several times until highly purified cells were obtained.Biological characters of BMSCs were observed.The surface antigens of BMSCs were identified by flow cytometry.The multipotentiality of BMSCs was assayed for differentiation into either osteoblasts or adipocytes.Results Homogeneous cells were obtained after BMSCs were passaged twice to thrice by Colony-Forming system.These BMSCs highly expressed positive surface antigens of BMSCs ( CD29 in 98.8％,CD90 in 98.4％ ) meanwhile seldom expressed negative surface antigens of BMSCs (CD31 in 2.6％,CD45 in 3％ ).BMSCs efficiently differentiated into osteoblasts and adipocytes.Conclusions Colony-Forming system is a simple and effective way to get highly purified BMSCs in a short time.

Objective To compare the difference of ELISA and LABScreen in detecting HLA antibodies and evaluate their effects on allograft rejection.Methods Consecutive patients undergoing kidney transplantion from November,2008 to December,2009 in the First Affiliated Hospital and the following up patients during the same period,with abnormal Scr who had completed kidney biopsies,were included in this study.Patients' HLA antibodies were detected by ELISA (Lambda antigen tray,LATTM) and LABScreen Mix Beads (LABScreen MIX,One Lambda,Canoga Park,CA,USA) or LABScreen Single antigen beads (LABScreenTM single antigen beads,One Lambda,Canoga Park,CA,USA).Patients' Scr were also detected at different time potints.Results There were 277 patietns included.Among them 145 (52.3％) cases were HLA antibody positive detected by LABScreen,which including 118 cases ELISA negative but LABScreen positive,and 27 cases both ELISA and LABScreen positive.No case was ELISA positive but LABScreen negative.Among 118 cases which were LABScreen positive but ELISA negative,41 (34.7％) cases happened acute or chronic rejection.However,only 24 cases happened rejection in 132 double negative cases (18.2％,P =0.003).There were 31％ patients in rejection group while only 12.8％ patients (P=0.01) in non-rejection group whose HLA antibody fluorescence intensity detected by LABScreen single antigen beads still increased two weeks after transplantation.Conclusion LABScreen is more sensitive than ELISA in detecting HLA antibodies,and its result highly correlates with the incidence of allograft rejection.

Aim To study the influence of estradiol on peripheral blood stem cells and angiogenesis in a rat model of myocardial infarction.Methods Acute myocardial infarction(AMI)model was bulit by ligation of left anterior descending coronary in ovariectomized SD rats.Then,they were treated with different doses of estradiol benzoate.CD34+ cells in myocardium at 24 h after AMI were detected by immunohistochemistry,and CD34+ cells in peripheral blood were measured by flow cytometry on day 1,3 and 7 after AMI.The expression of stromal cell-derived factor-1(SDF-1)in myocardium on day 1,3 and 7 after AMI and capillary density in myocardium at 28 days after AMI were evaluated by immunohistochemistry.Results CD34+ cells in peripheral blood and myocardium,the expression of SDF-1 and capillary density at 4 weeks after AMI in ovariectomized rats were obviously lower than those in ovary intact rats.However,after indication with different doses of estrogen,CD34+ cells in peripheral blood and myocardium,the expression of SDF-1 and microvascular density were increased in ovariectomized rats with AMI,this effect was strongest especially in estrogen replacement therapy group.Conclusions Estrogen could improve the mobilization and homing of bone marrow stem cells into myocardium,increase capillary density and improve prognosis after AMI,and this mechanism could be mediated by promotion of expression of SDF-1 in myocardium.Moreover,the effect of estrogen replacement before AMI was better than estrogen therapy after AMI.

Objective To investigate the effect of low nitrogen and low calorie parenteral nutrition combined with enteral nutrition support on inflammatory factors and immune function in patients with gastric cancer .Methods From May 2013 to May 2015,ninety patients with gastric cancer underwent surgical treatment in the Affiliated Hospital of Yanbian University were selected in the research ,and they were divided into control group and observation group according to different methods of nutritional support ,with 45 cases in each group.The control group received total parenteral nutrition support,the observation group was given low nitrogen and low calorie parenteral nutrition and enteral nutrition support, and supplemented by targeted nursing measures.The changes of levels of inflammatory factors and immune proteins in the two groups before operation and 7d after operation were compared.All patients were followed up for 6 months,and the quality of life and nursing satisfaction rate were compared between the two groups.Results At 7d after operation,the levels of immunoglobulin IgA ,IgM and IgG in the observation group were (2.62 ±0.43)g/L,(1.93 ±0.52)g/L,(14.58 ±3.32)g/L,respectively,which were significantly higher than those before operation [(2.01 ±0.30)g/L,(1.22 ±0.40) g/L,(12.42 ±5.64) g/L,t＝7.805,7.260,2.214,all P＜0.05] and those of the control group [(2.12 ±0.52) g/L,(1.53 ±0.41) g/L,(12.86 ±4.34) g/L,t＝4.971, 4.052,2.112,all P＜0.05].At 7d after operation,the levels of C reactive protein(CRP),tumor necrosis factor -α (TNF-α) in the observation group were (108.52 ±17.53)mg/L,(135.63 ±28.51)ng/L,respectively,which were lower than those before operation [(142.35 ±15.82) mg/L,(156.65 ±25.54) ng/L,t ＝9.611,3.684,all P＜ 0.05] and those of the control group[(135.68 ±14.54)mg/L,(145.52 ±27.53)ng/L,t＝8.000,3.824,all P＜0.05],the level of interleukin-2(IL-2) was higher than those before operation [(65.71 ±10.23)ng/L vs.(54.32 ± 7.83)ng/L,t ＝5.931,P ＜0.05] and that of control group [(57.62 ±5.86) ng/L,t ＝4.603,P ＜0.05].At 6 months after operation,the quality of life scores of the observation group were higher than those of the control group (t＝7.444,6.892,4.884,5.957,7.784,12.992,3.851,6.706,all P＜0.05).The nursing satisfaction rate of the observation group was 97.78%,which was higher than 82.22% of the control group ( χ2＝6.049,P ＜0.05). Conclusion Low nitrogen and low calorie parenteral nutrition combined with enteral nutrition support can effectively improve the immune function of patients with gastric cancer ,reduce the inflammatory response and ensure the quality of life of the prognosis,which is suitable for clinical application.

ObjectiveTo explore the effect of simvastatin on the level of serum adiponectin and asymmetric dimethylarginine (ADMA) in patients with acute stroke.MethodsNinety patients with acute stroke were selected and divided by random digits table method into common treatment group and intervention treatment group with 45 cases each.The volume of cerebral infarction and the evaluation score of clinical neurological impairment degree were calculated.The levels of serum adiponectin and ADMA before and after treatment were detected by erzyme-linked immunosorbent assay.The changes of serum adiponectin and ADMA before and after treatment in two groups were compared and their correlations with the volume of cerebral infarction and the evaluation score of clinical neurological impairment degree were analyzed.Results The level of serum adiponectin and ADMA in two groups before treatment had no statistical differences (P ＞ 0.05 ).The levels of serum adiponectin in two groups after treatment increased and ADMA decreased significantly,which had significant differences compared with those before treatment ( P ＜ 0.05 ).The level ofserum adiponectin in intervention treatment group was significantly higher than that in common treatment group and ADMA was obviously lower than that in common treatment group after treatment [(5.92 ± 0.15)mg/L vs.( 4.51 ± 0.13 ) mg/L,( 0.96 ± 0.13 ) μ mol/L vs.( 1.08 ± 0.15 ) μ mol/L ] ( P ＜ 0.05 ).Rank correlation analysis showed that the level of serum adiponectin before treatment was negatively correlated with the volume of cerebral infarction and the evaluation score of clinical neurological impairment degree ( r ＝ -0.75,-0.59,P ＜ 0.05).The level of ADMA before treatment was positively correlated with the volume of cerebral infarction and the evaluation score of clinical neurological impairment degree ( r =0.68,0.71,P ＜ 0.05 ).Conclusion Simvastatin can increase the level of serurm adiponectin and decrease the level of ADMA efficiently in patients with acute stroke,and improve the prognosis of the patients.

Objective To analyze the research hotspots and development trends in the pre-hospital emergency care field of stroke in China from 2014 to 2023 using bibliometric method.Methods Retrieve stroke pre-hospital emergency care research articles published in the China National Konwledge Infrastructure(CNKI),WanFang and CQVIP database from 2014 to 2023.Visualize and analyze authors,institutions,and keywords using CiteSpace 5.7.R5 software.Results We included 3,125 articles for analysis.The year 2021 marked the peak of publication activity with 500 articles.The top five authors with the highest number of publications were Guo Wei,Li Dou,Yu Longjuan,Peng Peng,and Zhang Lingjuan,respectively.The Beijing Emergency Center emerged as the institution with the most significant number of publications.Predominant research hotspots encompassed topics such as intravenous thrombolysis,therapeutic outcomes,nerve functions,pathways for emergency nursing care,and green Channel.Current trends are directed towards enhancing the quality of emergency services,optimizing interventional thrombectomy techniques,improving neurological outcomes,and prognostic assessments.Conclusion Over the last decade,there has been a progressive increase in the focus on pre-hospital emergency care research for stroke within China.Future research endeavors should aim at elevating the quality of emergency care,refining thrombectomy approaches,augmenting neurological functional recovery,and improving patient prognoses.

OBJECTIVETo evaluate the feasibility and safety of laparoscopic pancreaticoduodenectomy(LPD).

METHODSData of 66 patients from Sir Run Run Shaw Hospital undergoing LPD from September 2012 to September 2014 were reviewed. There were 44 male and 22 female with the mean age of (58.7±10.3) years and mean body mass index of (23.5±3.9)kg/m(2). Forty-five patients presented the symptoms and four of all had the history of abdominal surgery, including 2 cases of laparoscopic pancreatic surgery.

RESULTSOf 66 patients underwent laparoscopic procedure, 1 patient underwent LPD combined with right hepatic resection, 1 patient underwent laparoscopic distal gastrectomy with LPD, and 1 patient underwent LPD after laparoscopic distal pancreatectomy. The mean operative time was (367±49) minutes. The mean blood loss was(193±126)ml. The rate of overall postoperative complications was 36.4%(24/66), with 4.5%(3/66) of B or C pancreatic fistula and 7.6% (5/66) of bleeding. The mean postoperative hospital stay was (18.9±12.1) days. Mean tumor size was (3.8±2.3) cm, and the mean number of lymph nodes harvested was (20.3±10.9). Forty-severn patients were diagnosed as pancreatic adenocarcinoma(n=18), cholangiocarcinoma(n=7), ampullary adenocarcinoma(n=21), and gastric cancer(n=1), respectively.

CONCLUSIONLPD is feasible and safe under the skilled hand.

Adenocarcinoma ; surgery ; Aged ; Anastomosis, Surgical ; Female ; Humans ; Laparoscopy ; Length of Stay ; Male ; Middle Aged ; Operative Time ; Pancreatectomy ; Pancreatic Fistula ; pathology ; Pancreatic Neoplasms ; surgery ; Pancreaticoduodenectomy ; Postoperative Complications

Objective:To evaluate the biological characters of C57-TgN(HBV adr2.0)SMMU transgenic mice. Methods: Integration,expression,replication and histology change of hepatitis B virus gene in F6 transgenic mice were estimated by ge-nomic DNA PCR,Western blotting,ELISA,immunohistochemistry,serum DNA PCR,transmission electron microscopy and H-E staining. Results: Hepatitis B virus gene was integrated into F6 C57-TgN(HBV adr2. 0)SMMU transgenic mice and expressed HBsAg,HBcAg and X protein in liver tissue. HBsAg and HBeAg were expressed in serum of 19. 54% and 3. 39% F6 transgenic mice. Hepatitis B virus were replicated in serum and liver tissue of transgenic mice. Long-term integration,expression and replication of hepatitis B virus gene induced pathological lesion of transgenic mice liver and lung. Conclusion: C57-TgNCHBV adr2. 0)SMMU transgenic mice line has the biological characters including integration of hepatitis B virus gene into genomic DNA,expression and replication of hepatitis B virus gene in serum and liver, and histological change in liver and lung. It is a valuable animal system to study pathogenesis, treatment and prevention of hepatitis B virus.

Objective: To summarize the incidence and characteristics of postoperative complications after laparoscopic pancreaticoduodenectomy(LPD), and to share our experience on management of complications. Methods: The clinical data of 320 LPD performed by a single team in Sir Run Run Shaw Hospital and Zhejiang Provincial People′s Hospital between September 2012 and September 2017 were retrospectively analyzed, among which there were 196 males and 124 females with age of (60.2±11.6) years old.There were 306 patients who underwent standard LPD, and 14 patients who underwent extended LPD. The patients were divided into 2 groups of former 160 LPD and later 160 LPD according to the time order. By analyzing the differences of clinical outcomes between the two groups, especially focusing on the incidence of postoperative complications.The experience on management of complications was concluded. The prior surgical history of latter group was significantly higher than the former group(30.0%(48/160)vs. 18.8%(30/160), χ²=5.49, P=0.019), and the rest of baseline characteristics remained the comparable (P>0.05). For resectable lesions, LPD was performed by "No back" approach, following the principle of "From distal to cephalad, from ventral to dorsal, and from left to right" . As for the borderline resectable patients, LPD was performed by "Easy first" strategy. Student t test, χ² test or Fisher test was used to analyzed the data between the two groups respectively. Results: Of 320 LPD patients, 306 cases underwent standard LPD, 14 cases underwent LPD with resection of other organs.There were 278 LPD cases who followed "No back" approach, and 42 cases who followed "Easy first" strategy because of difficulty in creating the retro-pancreatic tunnel. And the overall morbidity was 32.2%(103/320) with reoperation rate of 5.3%(17/320). The perioperative mortality was 0.6%(2/320). The operation time of latter group was ((346.6±48.8)minutes), which was shorter than that of former group((358.0±54.4)minutes)(t=1.97, P=0.048). The blood loss of former and latter group remained comparable((207.9±135.8)ml vs.(189.6±121.4)ml, P=0.205). However, in subgroup analysis, the patients with blood loss less <200 ml of latter group decreased significantly from 59.4%(95/160) to 47.5%(76/160)(χ²=4.53, P=0.033). The overall morbidity of latter group was 28.8%(46/160), indicated a decrease from 35.6%(57/160) of former group without significant difference(P=0.188). Moreover, Grade A/B/C pancreatic fistula rate, Grade A/C bile leakage rate, Grade B/C postoperative hemorrhage rate of the later group tended to decrease, although they also didn′t reach a significant difference. However, the abdominal infection rate decreased significantly(χ²=3.93, P=0.047). The length of hospital stay remained comparable(P=0.156). Conclusions The most common complications after LPD were postoperative hemorrhage and pancreatic fistula. With specialized team and accumulated experience, the morbidity can decrease progressively by analyzing the leading cause and improving the technical skills.

Objective: To summary the experience of 233 cases of laparoscopic pancreaticoduodenectomy (LPD) performed by a single surgical team. Methods: Data of patients undergoing LPD from September 2012 to October 2016 were reviewed. There were 145 males and 88 females with the mean age of(60.3±13.0)years old, ranging from 19 to 92 years old, and the mean body mass index of (22.8±3.5)kg/m^2, ranging from 16.3 to 36.8 kg/m². There were 195 patients with clinical manifestation and 54 patients who had the history of abdominal surgery. Results: LPD were performed on 233 patients by same surgical team consecutively. The mean operative time was(368.0±57.4)minutes. Mean blood loss was(203.8±138.6)ml. The postoperative morbidity rate was 33.5%, with 6.9% of grade B or C pancreatic fistula and 9.9% of bleeding. The reoperation rate was 5.6%. The mortality during 30 days after operation was 0.9%. Mean postoperative hospital stay was (18.1±11.2)days. Mean tumor size was (3.9±2.4)cm, and the mean number of lymph nodes harvested was 21.3±11.9.One hundred and sixty-three patients were diagnosed as malignant tumor, including pancreatic adenocarcinoma(n=84), cholangiocarcinoma(n=17), ampullary adenocarcinoma(n=55), duodenal adenocarcinoma(n=5), gastric cancer(n=1)and duel cancer (n=1) located in distal stomach and duodenum. Conclusion The key point to make laparoscopic pancreaticduodenectomy a routine safe procedure is to operate the procedure under skilled hands in selected patients via suitable surgical approaches.