BACKGROUND: Vascular endothelial growth factor receptor 2 (VEGFR2) is primarily involved in vascular endothelial growth factor (VEGF)-mediated signal transduction and plays a critical role in the pathological angiogenesis that occurs in a number of diseases, including leukemia. Besides, VEGF secreted by leukemia cells also induces its own expression which leads to an enhanced production of VEGFR2 which contributes to the survival and proliferation of leukemia cells.OBJECTrVE: To evaluate the inhibitive effect of Lenti6/shVEGFR2 on the VEGFR2 expression and leukemia growth in mouse.DESIGN: A randomized, parallelized, controlled and open trial.SETTING: Department of Pediatrics, the Second Affiliated Hospital of Sun Yat-sen University; Biotechnology Research Center, Sun Yat-sen University.MATERIALS: The experiment had been done in the laboratories for Medical Research Center of the Second Affiliated Hospital, Sun Yat-sen University and Biotechnology Research Center, Sun Yat-sen University from May 2004 to January Lentiviral RNAi Expression System was purchased from Invitrogen, Co.,Ltd.; human VEGFR2 Mcb (PE) was purchased from R&D; CD31 immunohistochemistry kit was purchased from Boster, Co.,Ltd.; CD33-PE fluorescence labeled antibody was purchased from BD, Co.,Ltd.transiently and expression clone (Lenti6/shVEGFR2) was constructed, then cotransfected with ViraPowerTM Packaging Mix pU6/shVEGFR2 entry clone and transducting with Lenti6/shVEGFR2 expression clone, the effect on the development of intravenous xenograft leukemia mouse model, the distribution of microvessels in mouse bone marrow was observed after leukemia model mouse injected with recombinant lentivirus (group B); leukemia model mouse injected with recombinant lentivirus and endothelial cell (group C); leukemia model mouse injected with endothelial cell (group D). Through detecting changes of CD33 positive cells and microvessel density (MVD) in bone marrow, observing peripheral blood cell (PBC)smear and slice of liver, spleen, the effect of Lenti6/shVEGFR2 recombinant lentivirus on mouse leukemia was evaluated.mediated with lentivirus on VEGFNEGFR2 paracrine and autocrine loops in leukemia mouse.effective in inhibiting HL60 cell. pU6/shVEGFR2 entry clone constructed according to it had cell inhibitory rate as high as after transfection of pU6/shVEGFR2 entry clone and transduction of Lenti6/shVEGFR2 expression clone: 48 hours after transfection of pU6/shVEGFR2 entry clone and transduction of Lenti6/shVEGFR2 expression clone, the cell growth inhibitive rates were similar. However, the cell growth inhibitive rate of entry clone descended rapidly after 48 hours (P＜0.01); which of expression clone changed slowly, reaching the peak at 96 hours, dropped slightly, having no significance mouse: The amount of HL60 cells in bone marrow of groups A, B and C detected with flow cytometry were (25.8%±4.9)%, (14.3%±5.1)%, (8.4±2.6)%, respectively (P＜0.05); MVD in group C was obviously less than that in group D (P＜0.05); The amount of HL60 cells in leukemia model mouse injected with recombinant lentivirus and endothelial cell was the lowest as compared with the other groups.

Objective To investigate the optimal dose of apocynin to protect severe acute pancreatitis (SAP) and SAP caused intestinal injury in rats. Methods A total of 53 SPF male Wistar rats were randomly allocated into five groups:sham operation group (SO group, n=10), SAP group (n=12), low-dose apocynin group (25 mg/kg,n=11), medium-dose apocynin group (50 mg/kg, n=10) and high-dose apocynin group (100 mg/kg,n=10). SAP model was prepared by retrograde infusing 5%sodium taurocholate (1 mL/kg) into biliopancreatic duct of rat. At thirty minutes before modeling, apocynin was injected into rat to intervention. The survival condition was recorded at 12 h after modeling, and blood samples were obtained for detecting serum amylase (AMY), alanine aminotransferase (ALT) and creatinine (Cr). Pancreatic and ileal tissue samples were obtained for HE staining and pathological examination. Results Two rats died in SAP group and one died in low-dose apocynin group. The quantity of ascites, the levels of AMY, ALT, Cr and pancreatic and intestinal pathologic scores were significantly increased in SAP group than those in SO group (P<0.05). Except the levels of Cr and intestinal pathologic score, there was no significant difference between low-dose apocynin group and SAP group. The quantity of ascites ascites, levels of AMY, Cr and pancreatic and intestinal pathologic scores were significantly lower in medium-dose and high-dose apocynin groups than those in SAP group (P<0.05). The levels of ALT and Cr were significantly higher in high-dose apocynin group than those of medium-dose apocynin group (P<0.05). Conclusion Apocynin improves SAP symptoms and reduces SAP caused intestinal injury in rats, which may be related to the inhibition of NOX activity, and 50 mg/kg of apocynin is the optimal dose.

Objective To evaluate the curative effect of infection caused by peritoneal dialysis(PD) catheter. Methods Eighty patients with infections caused by peritoneal dialysis catheter were enrolled in this study and randomized into silver-containing hydrofiber dressing group and control group with 40 patients in each group. The control group was treated with iodophor and/or gentamicin and the treatment group with silver-containing hydrofiber dressings. The two groups were compared in terms of curative effect at the wounds and time for cure. Result The cure rate in the treatment group was significantly higher than that in the control group (P<0.05), and the cure time in the observation group was significantly shorter as compared to the control group. Conclusion Silver-containing hydrofiber dressing is effective in the treatment of PD catheter-induced infections , and it can shorten treatment duration and reduce the frequency of dressing change.

To establish a scientific and feasible evaluation index system and to assess medical research programs subsidized by the Shanghai Municipal Health Bureau in 2002-2006, so as to provide reference to aid the management department in decision-making and better guidance of research projects. Delphi expert consultation was employed to construct an evaluation index system, and used the system to comprehensively assess the funded projects. In the end, we found that the funding of the Shanghai Health Bureau had certain promoting effect in optimization of research team, startup of scientific research, training of research personnel, and the development of disciplines.

Objective To evaluate the effectiveness of low-dose erythromycin for the treatment of feeding intolerance in preterm infants in China.Methods In this study,random clinical trials on the treatment of feeding intolerance in preterm infants with intravenous low-dose erythromycin published were searched at Chinese Journal Full-text Database,Chongqing Weipu Database and Wanfang database by using the methods of Cochrane systematic review.At the same time the information from related journals,professional data and network were hand-searched.The publishing deadline for the literatures reviewed in this study was August 2012.Statistical analysis of clinical data was performed by using RevMan 4.2 software provided by the Cochrane Collaboration.Results A total of 9 studies were included.The results showed that compared with the group of comprehensive therapy,the group of low-dose erythromycin was superior in the following aspects with significant differences(P ＜ 0.05):the average length of hospital stay,time of parenteral nutrition,time to full feeding,the incidence rate of feeding intolerance (Z =3.44,P =0.000 6 ; Z =6.78,P ＜0.000 01 ; Z =3.96,P ＜ 0.000 1 ; Z =2.51,P =0.01).Conclusion Low-dose erythromycin therapy for feeding intolerance in preterm infants is superior to the comprehensive therapy.It provides a prospective therapeutic method for feeding intolerance in preterm infants.However,large scale,multicenter and well-designed clinical trials should be adopted to confirm the conclusions.

The methods of 1H Nuclear Magnetic Resonance (NMR) and mass spectroscopy were used in detecting the metabolites of brodimoprim (BDP) in rat plasma. The endogenic compounds in the plasma were removed with solid phase extraction (SPE) column firstly, then the mixture of metabolites was identified with 1H NMR and mass spectroscopy (MS). Two metabolites of BDP in the plasma at 20h were detected, they were demethyl-brodimorpim glucuronide and brodimoprim sulfurate. The study proved that the method of SPE coupled with NMR and MS can be applied to the analysis of metbolites in plasma quickly and conveniently.

BACKGROUND: Previous studies have confirmed that embryonic stem cells call be induced to differentiate into insulin-producing cells, but the induction process takes a long time. Most of the processes take about one month.OBJECTIVE: Activin A, all-trans retinoic acid (ATRA), basic fibroblast growth factor (bFGF) and nicotinamide were applied in vitro in combination to observe whether mouse embryonic stem cells could be induct to differentiate into insulin-producing cell in a relatively short time.DESIGN: Cell observation experiment.SETTING: Shanghai Institute of Endocrine and Metabolic Diseases, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.MATERIALS: This study was performed at Shanghai Institute of Endocrine and Metabolic Diseases from October 2004 to February 2006. Two mice of clean grade and of 12.5-14.5 days of gestational age were provided by Shanghai SLAC Laboratory Animal Co., Ltd (Permission No. 2004A034). The protocol was performed in accordance with ethical guidelines for the use and care of animals. Mouse embryonic stem cell lines were supplied by Dr Changxian Zhang (CNRS UMR5641, France). Activin A was the product of the R & D Corporation. ATRA and nicotinamide were supplied by the Sigma Corporation, USA. BFGF was supplied by Gibco Corporaion. METHODS: Head and viscera were removed from embryos of the pregnant mouse. The remaining tissues were cut into pieces and digested with trypsin. Cell suspension was centrifuged and inoculated at 3×108L-1. The cells could be used as mouse feeder layer after 2-3 times of passage. The mouse embryonic stem cells (ESCs) were inoculated onto the feeder layer in knockout Dulbecco's modified Eagle medium (DMEM) supplemented with leukemia inhibitory factors (LIF). ESCs were passaged at 1:3-1:6 after 2-3 days of culture. Culture medium with serum was added into the culture dishes to terminate the digestion. Cell fluid was centrifuged and supernatant was discarded. The sediments were prepared into suspension and inoculated at 2.5×104 with LIF-free culture medium. After 24-48 hours, embryonic bodies (EBs) were collected and replated in 1% Matrigel-coated dishes. When began to adhere to the dishes, EBs were cultured in 10% FBS/DMEM supplemented with 100μg/L activin A for 24 hours. Then EBs were switched to 10% FBS/DMEM for 6-8 hours as an interval. After this interval. EBs differentiated were cultured in 10% FBS/DMEM with 10<-6mol/L RA for another 24 hours followed by culture in 10% FBS/DMEM supplemented with 10μg/L bFGFs for 3-5 days. Finally, EBs differentiated were cultured in DMEM/F12 supplemented with N2 supplement, B27 supplement, 1μg/L laminin, 10μg/L bFGFs, and 10mmol/L nicotinamide for 3-5 days. Dithizone (DTZ) staining, inununofluorescent staining and reverse transcription-polymerase chain reaction (RT-PCR) were applied to detect insulin expression in the differentiated cells.MAIN OUTCOME MEASURES: Induction of ESCs, DTZ staining and immunofluorescent staining as wel as RT-PCR detection.RESULTS: Mouse ESCs growing on a feeder layer formed many colonies with clear boundary and dense structure. However, there was no obvious outer limit between these ESCs. EBs began to adhere to the dishes, which were coated with matrigel, on the 2nd day. After activin A and ATRA interval induction, EBs spread, and most of the living cells were epithelial cell-like when cultured in 10% FBS/DMEM supplemented with 10μg/L bFGFs. After culturing in DMEM/F12 supplemented with N2, B27, nicotinamide, bFGFs and laminin, the cells formed small clusters. The insulin-producing cells were stained dark red with DTZ, and the cells stained with primary antibody to insulin were insulin-positive. After 2 weeks of induction of activin A, ATRA, bFGFs and nicotinamide, the insulin-producing cells expressed insulin 2, Pdxl, Nkx6.1, Nkx2.2, PP, IAPP, Glut2, Somastatin, Hnf3β and Neuro D mRNA but did not express insulin 1 mRNA.CONCLUSION: Mouse ESCs call be induced to differentiate into insulin-producing cells by activin A, ATRA, bFGFs and nicotinamide in vitro. Induction time call be shortened to 2 weeks.

Objective To investigate the effects of constant magnetic field (CMF) on proliferation, apopto-sis and nitric oxide (NO) secretion of rat bone marrow-derived endothelial progenitor cells (EPCs) intervened by C-reactive protein (CRP). Methods EPCs were isolated from rat bone marrow by density gradient centrifugation and cultured on fibronectin-coated dishes. The cells were divided into five groups, i. e., control group, CRP (12 μg/ml) group, CRP plus CMF (0.1, 0. 5, 1.0 mT) groups. Samples were collected 24 hours after incubation. Cell proliferation was measured by MTT chromatometry. Apoptosis rate was detected by flow-cytometry. NO content of culture medium was measured by nitrate reductase method. Results As compared with control group, cell prolifer-ation in CRP group reduced significantly (0. 265±0. 008 vs 0. 316±0. 011, P < 0.05), NO secretion also de-creased significantly [(22.7±4.5) μmol/L vs (37.6±3.8) μmol/L, P < 0.05], cell apoptosis rate elevated sig-nificantly [(10.8±0. 8) % vs (4.2±0.5)% ,P < 0.05]. Cell proliferation in CRP plus 0. 5 mT or 1.0 mT CMF group (0. 295±0. 009,0. 302±0. 010) were much more than those in CRP group (P<0.05), NO secretion contents [(28.3±4.9) μmol/L, (29.2±5.6) μmol/L]were also much more than those in CRP group (P < 0.05) , apopto-sis rate [(7.4±0.5)% ,(6.9±0.6)%]was significantly lower than that in CRP group (P <0.05). Conclusion CMF at intensity of 0.5 mT and 1.0 mT can antagonize the effects of CR, promote proliferation of EPCs and secretion of NO and inhibit apoptosis rate of EPCs.

Objective To study the risk factors related to recurrence of gastrointestinal stromal tumor (GIST) after discontinuing postoperative adjuvant imatinib mesylate (IM) treatment.Methods We retrospectively analyzed our clinical database of 138 GIST patients who received radical resection and subsequent IM adjuvant treatment at the Renji Hospital,Shanghai Jiaotong University School of Medicine between January 2006 and January 2014.Results For the entire Multivariate analysis study group,the overall 5-year recurrent free survival (RFS) rate was 54.5％.There were two tumor characteristics which were independent prognostic factors of GIST treated by postoperative IM:Ki67 index (P =0.005),and serosal invasion (P =0.026).The accuracy of comprehensive evaluation based on the two weighted variables was better than NIH staging criteria(AUC:0.714 vs.0.631).Furthermore,two risk groups were created according to the risk model with 5-year RFS of 81.3％ and 31.1％ as low-risk and high-risk groups,respectively (P ＜0.05).Conclusions For patients with intermediate or high risk in NIH classification,if there was tumor serosal invasion,or if there was no local invasion but Ki67 index ＞ 8％,extended continuous IM adjuvant treatment should be recommended after the primary tumor was radically resected.

Background and purpose:The young breast cancer patients were treated with goserelin without individualized regimen, and lack of available clinical marker. The aim of this study was to investigate the role of anti-Müllerian hormone (AMH) in evaluation of individualized treatment of ovarian function suppression in the young breast cancer patients.Methods:Forty-one young patients with estrogen receptor (ER) and progesterone receptor (PR) positive breast cancer from May 2012 to Jan. 2014 were randomly divided into 2 groups to undergo radical resection of breast cancer. According to postoperative treatment, one group was treated with goserelin + chemotherapy (n=20), and the other group received chemotherapy alone (n=21). Thirty female patients in the same age group were selected as normal control group. The time of menopause and menstrual recovery after the goserelin + chemotherapy or chemotherapy alone were observed in 2 groups. In early follicular phase (day 3-5) of the cycle preceding the operation and 3, 6 courses after the goserelin + chemotherapy treatment or chemotherapy treatment, serum levels of AMH, FSH and E2 were measured in 2 groups. Accordingly, serum levels of AMH, FSH and E2 were evaluated as well in normal control group.Results:There were no signiifcant differences in preoperative general conditions and preoperative serum FSH and E2 levels among the 3 groups (P>0.05). Compared with normal control group, the preoperative serum AMH levels of young breast cancer patients were decreased signiifcantly (P=0.04). The menopause time and menstrual recovery time in 2 chemotherapy groups were signiifcantly shorter than that in normal control group (P=0.00). Compared with normal control group and preoperative measurement, the differences in serum FSH and E2 levels were not statistically significant in goserelin + chemotherapy group or chemotherapy alone group (P<0.05). The serum AMH levels measured at different time points of the goserelin + chemotherapy group and chemotherapy alone group were decreased signiifcantly (P<0.05). Compared with the chemotherapy group, the serum AMH levels of the goserelin + chemotherapy group after 6 courses were signiifcantly decreased, and then signiifcantly increased 6 months after menstrual recovery (P<0.05).Conclusion:This study demonstrated that the serum AMH levels were obviously decreased after the ovarian function suppression treatment and increased after the menstrual recovery compared with evaluation of other ovarian reserve index. The serum AMH level could suggest ovarian reserve damage even after ovarian function has recovered to the noticeable level. Thus, AMH could be used clinically to evaluate the ovarian reserve of breast cancer patients as a potential marker for the individualized ovarian function suppression treatment in young breast cancer patients.