Blood-brain barrier(BBB)is mainly composed of brain microvascular endothelial cells(BMECs).According to the special cell membrane transport system and the extensive tight junctions,the brain microvascular endothelial cells play an important role in the CNS.Also,there is a close relationship between BMECs and the cerebrovascular diseases,such as cerebral ischemia,cerebral edema,and so on.Therefore,the culture of brain microvascular endothelial cells provides a new tool for studying the cerebrovascular diseases in vitro.The researches in recent years have been reviewed in this test,approaching the cultivation of the rat brain microvascular endothelial cell from the segregation,purification and culture in turns,in order to provide a foundation for the studies of the brain microvascular endothelial cell;in vitro.

Objective To investigate the effects ofLiuwei-Dihuang Pills combined with Ginkgo Biloba Leaves Extract Tablets on diabetic complications in patients with type 2 diabetes mellitus (T2DM). Methods Seventy-eight patients with T2DM were randomized into a TCM treatment group and a conventional treatment group by random number table method, 39 in each group. The patients in the conventional treatment group were regularly using antihyperglycemic drugs or insulin. On the basis of conventional treatment group, the patients in the TCM treatment group were additionally treated withLiuwei-Dihuang Pills and Ginkgo Biloba Leaves Extract Tablets. Intima-media thickness (IMT) of internal carotid artery, vibration perception threshold (VPT) and diabetic retinopathy in both groups were detected before the treatment, 12 and 36 months after the treatment.Results The VPT in the left foot 12 months after the treatment in the TCM treatment group were significantly lower than that in the conventional treatment group (13.98 ±4.38 Vvs. 18.70 ±2.43 V;t=2.764, P=0.008). IMT of internal carotid artery (left: 0.81 ± 0.16 mmvs.0.70 ± 0.10 mm,t=3.120,P=0.003; right: 0.81 ± 0.17 mmvs. 0.73 ± 0.12 mm,t=2.286,P=0.026), VPT (left foot: 14.82 ± 6.45 Vvs. 20.63 ± 9.75 V,t=2.714, P=0.009; right foot: 16.73 ± 7.10 Vvs. 20.73 ± 10.35 V,t=2.001,P=0.048) and incidence of diabetic retinopathy (35.9%vs. 53.8%;χ2=5.804,P=0.016) 36 months after the treatment in the TCM treatment group were significantly lower than those in the conventional treatment group.ConclusionsLiuwei-Dihuang Pills combined with Ginkgo Biloba Leaves Extract Tablets can effectively reduce the progression of diabetic complications in patients with T2DM.

Objective To compare postoperative adjuvant chemoradiotherapy with adjuvant chemotherapy in patients with gastric cancer by a meta-analysis.Methods PubMed,EMbase,Cochrane Library,Wanfang,CNKI,VIP,and CBM databases were searched to identify the controlled clinical trials of postoperative adjuvant chemoradiotherapy versus adjuvant chemotherapy for gastric cancer.The obtained data were analyzed using RevMan 5.2.5 and Stata 12.0.The difference between two groups was estimated by calculating the odds ratio (OR) with 95％ confidence interval (CI).Results A total of 12 controlled clinical trials involving 1674 gastric cancer patients,which were selected according to inclusion and exclusion criteria,were included in this meta-analysis.The meta-analysis showed that the 3-and 5-year survival rates were significantly higher in the adjuvant chemoradiotherapy group than in the adjuvant chemotherapy group (OR=2.96,95％ CI=1.75-5.03,P=0.000; OR=1.45,95％ CI=1.06-1.99,P =0.020);the local recurrence rate was significantly lower in the adjuvant chemoradiotherapy group than in the adjuvant chemotherapy group (OR =0.50,95％ CI =0.34-0.72,P =0.000) ; there was no significant difference in distant metastasis rate between the two groups (OR =0.79,95％ CI =0.58 -1.07,P =0.130).Conclusions The meta-analysis of existing study results shows that compared with adjuvant chemotherapy alone,adjuvant chemoradiotherapy is a relatively safe and effective postoperative treatment for gastric cancer.

Objective : To explore the influencing factors for maternal anemia during the third trimester of pregnancy, so as to provide the evidence for the effective prevention of anemia. Methods : Data of parturients, who received prenatal care and delivered at Hangzhou Women＇s Hospital from October to December of 2019 were retrospectively collected and analyzed. The obstetric electronic medical record system of this hospital was used to collect general information of the subjects. Serum ferritin, serum vitamin B12, serum folic acid and hemoglobin were determined at the second and third trimester. A multivariate logistic regression model was used to analyze the influencing factors for anemia in the third trimester. Results : A total of 1 143 parturients were enrolled, with a median age of 29 ( interquartile range: 4 ) years. Among them, 1 050 cases had normal weight before pregnancy, accounting for 91.86%; 267 cases were multiparas, accounting for 23.36%; 74 cases of anemia occurred during the third trimester of pregnancy, accounting for 6.92%.Multivariate logistic regression analysis indicated that the multipara（OR=1.714,95%CI: 1.032-2.848）, iron deficiency duiring the second trimester（OR=3.301,95%CI: 1.401-7.781）, iron deficiency during the third trimester （OR=14.134,95%CI: 1.944-102.774）and folate deficiency during the third trimester（OR=2.960,95%CI: 1.295-6.766）were influencing factors for anemia during the third trimester. Conclusion Anemia during the third trimester of pregnancy is related to iron deficiency during the second and third trimesters, folic acid deficiency during the third trimester, and multipara.

Objective To investigate the short-term clinical efficacy of levosimendan on treating patients with decompensated cardiac insufficiency.Methods One hundred and twenty patients with heart failure (NYHA Ⅲ-Ⅳ or Killip Ⅲ) were randomly divided into levosimendan group(n =60) and control group(n =60).The patients in levosimendan group were given intravenous levosimendan for 24 hours beside conventional heart failure medications.The patients in control group were given the conventional heart failure medications.The left ventricular ejection fraction (LVEF) was recorded and B-type natriuretic peptide (BNP) were measured before and after treatment.NYHA grade and mortality also were recorded.All patients were followed up for 3 months.Results The LVEF in the levosimendan group after the treatment was (35.6 ± 13.3)％,significantly higher than that in the control group ((31.4 ± 6.7) ％,F =8.952,P =0.002).The BNP in two groups after treatment were lower compared with before treatment(P ＜0.05).And it was more remarkable after treatment in levosimendan group compared with control group (441.0 (212.5,1050.0) ng/L vs.870.0 (435.0,1267.0) ng/L,P =0.014).The change of NYHA grade in levosimendan group was better than that in control group after 5 d.The recovery rate and ineffective or deterioration rate in levosimendan group were 45.0％ (27/60),26.7％ (16/30) and 43.3％ (26/60) respectively,higher than that of control group (28.3％ (17/60),20.0％ (12/60),36.7％ (22/60)) (OR =2.280,95％ CI 1.163-4.468,P =0.016).There was no significant difference in term of mortality between in hospital and 3 months follow-up in the levosimendan and the control group (20％ (12/60) vs.25％ (15/60),28.3％ (17/30) vs.41.7％ (25/60),x2 =1.543,P =0.214 and x2 =2.590,P =0.108).There was a decreasing trend regarding of readmission rate during 3 months in levosimendan group compared with that of the control group (21.7％ (13/60) vs.33.3％ (20/60),x2 =3.591,P =0.058),but mortality or readmission rate was lower than that in the control group (46.7 ％ (28/60)vs.66.7％ (40/60),x2 =4.835,P =0.028).Conclusion The short-term clinical efficacy of levosimendan on treating patients with decompensated cardiac insufficiency is remarkable better than the traditional treatment.

Objective To evaluate laparoscopic radical cholecystectomy for T2 gallbladder cancers.Methods A retrospective analysis was made on 44 patients with pathology confirmed T2 gallbladder cancer undergoing laparoscopic vs open radical cholecystectomy.The clinicopathological and follow-up data were compared.Results Laparoscopic cholecystectomy was performed in 32 patients,the implantation metastasis rate of this approach has no statistical differences compared with the open cholecystectomy (P =1.000).26 patients underwent laparoscopic radical resection and the remaining 18 patients underwent open radical resection.There was no statistical difference in operation time (P =0.953),blood loss (P =0.193)and postoperative complications (P =1.000),but the laparoscopic radical resection group is superior to the open group on postoperative pain grading (P =0.022),ambulation time (P =0.000),nothing per mouth time (P =0.000) and length of hospital stay (P =0.048).The mean number of lymph nodes retrieved was 5 ±4 (range 1-12) in the laparoscopic radical group and 6 ± 3 (range 1-12) in the open group (P =0.983);the 1,3,5 year survival rates was 92.3％,70.3％,61.5％ and 92.3％,76.3％,69.3％,respectively(P =0.473).Conclusions Initial laparoscopic cholecystectomy does not increase the rate of seeding metastasis in the context of intact gallbladder carcinoma.Totall laparoscopic radical resection is feasible in selected T2 gallbladder cancer patients.

Objective To evaluate the short-term clinical efficacy and safety of administration of levosimendan or milrinone added to conventional therapy in patients with decompensated heart failure.Methods A total of 180 patients admitted due to heart failure [NYHA (New York Heart Association) class Ⅲ or Ⅳ] were randomly (random number) divided into control group,milrinone group and levosimendan group (n =60,each group).A continuous infusion of milrinone added to conventional therapy was administered for 72 hours in milrinone group,while administration of levosimendan for 24 hours in levosimendan group.The changes in left ventricular ejection fraction (LVEF),left ventricle end-diastolic diameter (LVDD) and B-type natriuretic peptide (BNP) plasma level were compared between before and after treatment,respectively,and comparisons of improvement in cardiac function (NYHA class) and hospital mortality were carried out among three groups.Patients were further followed up at 3 months after treatment.Results The LVEF in levosimendan group after treatment had significantly more increased than that in control group [(32.0±6.3)％ vs.(30.6 ±5.5)％,P =0.007].Compared BNP before treatment,the sums of BNP deducted were 444.0 (-74.0,1068.0) pg/mL,469.0 (141.5,1151.5) pg/mL and 936.5 (437.8,1566.8) pg/mL in control group,milrinone group and levosimendan group,respectively after treatment (all P ＜ 0.01).Moreover,the deduction in BNP was more dramatic in levosimendan group compared with control or milrinone group (t =3.256 or 2.665,P =0.004 or 0.026).After treatment for 5 days,the probability at least of achieving more effectively better improvement in NYHA class (cardiac function) in levosimendan group was 2.036 times that of control group (95％ CI:1.030-4.028,P =0.041).The incidence of combined end point events (death or readmission) in levosimendan group was significantly lower than that in milrinone group (50％ vs.70％,HR =0.573,95％ CI:0.358-0.917,P=0.020),while in hospital mortality,readmission or 3-month mortality incidence was similar among 3 groups (P ＞ 0.05).Conclusions The short-term clinical efficacy of levosimendan is superior to that of milrinone or conventional therapy in patients with decompensated heart failure.

Objective To investigate the distribution of eosinphililic neurons ( ENs),reactive astrocytes ( RAs),and infarction after transient cerebal ischemia,and the time profile of pathomorphological changes.Methods Unilateral forebrain ischemia was induced in Mongolian gerbils by two 10 minutes unilateral common carotid artery occlusions with a 5 hours interval.Laser Doppler flowmetry was used to detect intra-ischemic anterior cortex blood flow.Animals were sacrificed at 24 hours,4 days,2 weeks,4 weeks,16 weeks and the brain were prepared for pathomorphological assay.Results Intra-ischemic laser Doppler flowmetry show significant ischemia during carotid artery occlusion:22.1％ ± 9.5％,26.3％ ± 4.9％,37.5％ ± 3.5％,F =67.219,P ＜ 0.01 ; the decrease was significantly greater in the anterior cortex.ENs appeared in middle and deep layers at 24 hours postischemia,and ENs area extend to all layers of cortex by 4 days.Large areas of high EN density ( ≥80/mm2) evolved to infarcts between 4 days and 4 weeks.Posterior cortex evolved to low EN area ( ＜ 80/mm2) without transformation into infarcts.RAs were consistently distributed in areas with ENs,and RA areas with high EN density were largely transformed into infarcts between 4 days and 4 weeks. Delayed astrocytic death took place in the RA areas with high EN density.Conclusion Density of ENs is an important indicator of delayed astrocytic death and infarction in postischemic tissue.

BACKGROUND:Under the in vitro conditions of cell harvesting, culture, and transplantation, whether bone marrow stromal cells (BMSCs) can be effectively applied in local gene therapy remains unclear.OBJECTIVE: To construct a recombinant eukaryotic expression plasmid carrying human bone morphogenetic protein-7 (hBMP-7) gene, and to expect to enhance osteoinductive properties of rabbit BMSCs transfected.DESIGN, TIME AND SETTING: A cell-genomics in vitro observation was performed at the Laboratory of Scientific Research, Second Affiliated Hospital of Harbin Medical University between July 2006 and July 2007.MATERIALS: Human healthy fresh placental tissue was provided by the Department of Gynaecology and Obstetrics, Second Affiliated Hospital of Harbin Medical University. Written informed consent was obtained from the women. One healthy male New Zealand rabbit was provided by the Laboratory Animal Center, Harbin Medical University.METHODS: hBMP-7 gene was cloned from human placental tissue to construct a recombinant eukaryotic expression plasmid carrying hBMP-7 gene by conjugating with eukaryotic expression vector pcDNA3.1. BMSCs were isolated from rabbit bone marrow and cultured in vitro. Then they were divided into 3 groups: pcDNA3.1-hBMP-7-transfected, pcDNA3.1 -transfected, and untransfected. 5×106 BMSCs were inoculated into a 60 mm3 flask containing antibiotic-free medium 1 day prior to transfection.MAIN OUTCOME MEASURES: RT-PCR and immunohistochemistry were employed to detect hBMP-7 expression in BMSCs, alkaline phosphatase activity, hydroxypreline content, and osteocalcin production in each group. RESULTS: After 72-hour transfection, a 1.3 kb fragment was seen in the pcDNA3.1-hBMP-7-transfected group, showing brown granules in the endochylema, but not seen in the pcDNA3.14ransfected and untransfected groups. ALP activity in the pcDNA3.1-hBMP-7-transfected group significantly increased at 2 days after transfection, peeked at 8 days, and still increased at 10 days. At each time point, alkaline phosphatase activity, hydroxyproline content, and osteocalcin production were significantly higher in the pcDNA3.1-hBMP-7-transfected group than in the pcDNA3.1 -transfected and untransfected groups (P<0.05 or P<0.01).CONCLUSION: Recombinant eukaryotic expression vector pcDNA3.1- BMP-7 was constructed successfully. Results indicated that hBMP-7 was expressed in BMSCs sufficiently and was involved in inducing differentiation of BMSCs into osteoblasts. The method would provide substantial basement for hBMP-7 gene therapy.

Objective To evaluate the improvement effect of levosimendan by vein injection on short term cardiac function of patients with decompensated heart failure.Methods One hundred and sixty patients admitted due to heart failure were randomly divided into levosimendan group and control group (80 subjects for each group).Patients in control group were given a regular therapy including diuretics,vasodilators (including the recombinant human brain natriuretic peptide),angiotensin converting enzyme inhibitor(ACEI) or angiotensin Ⅱ receptorantagonists(ARB),β blockers,spironolactone and stain.Patients in levosimendan were administered levosimendan for 24 hours plus regular therapy.The improvements of dyspnoea in 9 days and cardiac function classification in 30 days after therapy were assessed.Mortality of 1 month and 3 month in two group were calculated and compared during follow-up.Results The dyspnoea improvement rate was superior than that of control group during 9 days (OR =1.956,95％ CI:1.156-3.310,P =0.013).The improvements in the levosimendan group were better than in the control group at 1 st day (OR =2.261,95 ％ CI:1.280-3.999,P =0.005),at 3rd (OR =2.002,95 ％ CI:1.111-3.607,P =0.021) and 5th day (OR =1.846,95 ％ CI:1.009 -3.377,P =0.047).However,there was no significant difference in term of improving dyspnoea between the levosimendan group and the control group at 9th day (P =0.126).Similarly,the improvement of cardiac function classification in the levosimendan group was superior than the control group during 30 days (OR =1.933,95％ CI:1.229-3.040,P =0.004).Although no significant difference was seen regarding of improving cardiac function classification between the two groups at 30th day after treatment (P =0.115),the improvements in the levosimendan group were better than in the control group at 3rd (OR =1.986,95％ CI:1.195-3.300,P =0.008),5th (OR =2.268,95 ％ CI:1.329-3.873,P =0.003),9th (OR =2.627,95 ％ CI:1.419-4.860,P =0.002) and 14th day(OR =2.212,95％ CI:1.189-4.112,P =0.012).Moreover,there was a nonsignificant reduction in terms of mortality in levosimendan group during 1-month and 3-month follow-up compared with control group (P ＞ 0.05).Condusion Levosimendan can effectively improve the short-term cardiac function in patients with decompensated heart failure.