Objective To study the effects of octreotide on the proliferation and apoptosis of hepatocellular carcinoma cells, and provide an experimental basis for clinical application. Methods The proliferation of hepatocellular carcinoma cells (HepG2) was assessed by MTT and growth curve respectively, the contents of AFP in the culture supernatant were determined by eletrochemiluminescence immunoassay, and apoptosis was detected by fluorescent staining, transmission electron microscopy and flow cytometry. Results The proliferation of HepG2 was inhibited significantly by octreotide with a dosage dependant manner(range from 0.005 to 80 ?g/ml, P

Objective To investigate the protective mechanism of ischemic preconditioning on ischemic/reperfusion injury after pancreas transplantation in rats.Methods The model of diabetic SD rat was established. Twenty-four diabetic SD rats were randomly assigned to ischemic/reperfusion group (I/R group, n=6) and ischemic preconditioning group (IPC group, n=18). The rats in group IPC were averagely assigned to 3 sub-groups: group IPC_ 1 (5 min ischemic and 5 min reperfusion), IPC_ 2 (5 min ischemic and 5 min reperfusion twice) and IPC_ 3 (5 min ischemic and 5 min reperfusion thrice). Six normal SD rats whose abdomen was opened only served as control group, and they did not receive pancreas transplantation. I/R group and IPC group received pancreas transplantation. The superoxide dismutase (SOD) and myeloperoxidase (MPO) activity of grafts were monitored 2 h after reperfusion, the apoptotic cells in grafts were observed by TUNEL method, and the expression of Bcl-2 and Bax gene of the grafts was detected by Western blot.Results As compared with I/R group, the SIOD activity and the expression of Bcl-2 gene of grafts were significantly increased, while MPO activity, apoptotic index and the expression of the Bax gene in the grafts were markedly reduced in IPC group (P

Objective To probe into the optimal concentration of Wnt-11 to induce the differentiation of bone marrow mesenchymal stem cells (BMMSCs)into cardiomyocyte-like cells in vitro.Methods BMMSCs were isolated from the bone marrow of SD rats using whole bone marrow culture method.After cultured for 48 h, BMMSCs of the second generation were utilized for directed induction.Based on the final concentration of Wnt-11 , BMMSCs were divided up into Group A (100 ng/mL),Group B (200 ng/mL),Group C (400 ng/mL)and Group D (blank control).After 72-hour induction,the cells were cultured in complete medium for 4 weeks while cells in Group D were cultured only in the complete medium.The morphological changes were observed under the phase contrast microscope.Surface antigen expression of BMMSCs was identified by flow cytometry.When cells were cultured for 4 weeks,the expressions of Desmin,Connexin43 and cTnI were detected by immunocytochemistry. Meanwhile, the ultrastructural changes were observed using transmission electron microscope. The mRNA expressions of cardiac transcription factors GATA-4,Nkx2.5 andα-MHC in BMMSCs were detected by RT-qPCR at 1,2 and 4 weeks after induction.Results Primary BMMSCs formed cell colonies at 2 weeks;the cells were mainly fusiform or star-shape,and a few irregularly-shaped ones were also visible.The passaged cells were larger than those of primary culture.After induction,the cells exhibited long shuttle-shape and were aligned in parallel. Flow cytometery displayed that the positive rate of the surface antigens of BMMSCs CD29,CD45,and CD90 was 97.9%,0.4% and 99.5%,respectively.When BMMSCs-induced via Wnt-11 were cultured for 4 weeks,Desmin, cTnI and Connexin43 were all positively expressed in induction groups.Whereas in the blank control group they were slightly positive or negative;the positive rate in Group B was the highest (P<0 .05 ).Transmission electron microscopy exhibited that organelles such as rough endoplasmic reticulum,mitochondria,as well as some ribosomes were visible in the cytoplasm of these cells in each induction group.In addition,myofilaments were arranged in parallel in the cytoplasm.The cells in induction groups could express GATA-4 and Nkx2 .5 in the first week,and then the expression of them decreased in the second week,but then increased in the fourth week;gene expression in induction Group B was significantly higher than in the other two induction groups (P<0 .05 ).The expression of GATA-4 and Nkx2 .5 in Group D was 1 ,α-MHC was not expressed in the four groups during the culture period. Conclusion Wnt-11 can induce the differentiation of BMMSCs into cardiomyocyte-like cells in vitro,and the optimal concentration of Wnt-11 is 200 ng/mL.

Objective To explore the effect of recombinant pEGFP-N3-APC vectors carrying various APC functional domains on the expression of β-catenin in human colorectal cancer cells HT-29.Methods The recombinant plasmids were transfected into HT-29 cells mediated by lipofectamine~(TM) 2000, and detected by green fluorescence and RT-PCR. Western blot was applied to detect β-catenin expression level in HT-29 cells after transfection, and gray scales of electrophoresis strips were analyzed by SPSS 13.0.Results Green fluorescence and RT-PCR made clear that all 5 recombinant plasmids were successfully expressed in HT-29 cells. Western blot showed that β-catenin expression level in HT-29 cells was not affected after being transfected with pEGFP-N3-APC1, pEGFP-N3-APC2 and pEGFP-N3-APC3, and was distinctly affected after being transfected with pEGFP-N3-APC4 and pEGFP-N3-APC5, especially the later one. Conclusion The selected APC5 gene fragment with 15-amino acid repeats and SAMP repeats, which is relatively short, can degrade β-catenin level in HT-29 cells and may be applied in the gene therapy.

BACKGROUND:Ischemia/reperfusion (IR)injury during the pancreas transplantation can cause numerous postoperative complications, among which,secondary pancreatitis can cause small intestinal mucosal injury and result in severe Consepuence.OBJECTIVE:To observe the protective effect of ischemic preconditioning (IPC) on small intestinal mucosal barrier after pancreas transplantation in rats.DESIGN:Randomized controlled animal trial.SETTING:Department of Gastrointestinal Surgery,Xijing Hospital,Fourth Military Medical University of Chinese PLA.MATERIALS:This trial was done in the Laboratory of Gastrointestinal Surgery,Xijing Hospital,Fourth Military Medical University of Chinese PLA between September 2001 and April 2004.Eighty-three male SD rats were involved in this trial.METHODS: Forty-seven rats were randomly chosen to prepare diabetic rat models by penile-intravenous injection of 65 mg/kg streptozotocin.Thirty-six successful model rats were randomized into 3 groups,with 12 in each group:IR group,donor IPC(DIPC)group and recipient with two hindlims IPC(RIPC)group.Twelve of the remaining 36 normal rats served as control group,and the other 24 rats were used as donors.Laparotomy was conducted only in control group,and pancreas transplantation was conducted in the other 3 groups In DIPC group,the splenic vessels of donors were blocked for 5 minutes and reperfused for 5 minutes twice before obtaining pancreas from donor;In the RIPC group, blood flow of two hindlimbs of recipients was blocked for 5 minutes and reperfused for 5 minutes before reperfusing the pancreas of donor,and this procedure was repeated 3 times.IR group was untouched.MAIN OUTCOME MEASURES:① On the 5th day after operation,6 rats were randomly chosen from each group to detect small intestinal permeability[expressed with plasm fluorescent-isothiocyanate-dextran(FITC-dextran)concentration]and absorption function(expressed with plasm xylose concentration).② On the 5th day after operation.blood was taken from the left 6 rats in each group to detect serum tumor necrosis factor-α(TNF-α) and nitric oxide(NO)level as well as superoxide dismutase(SOD)and amylase activity.Ileal mucosal tissue was taken to detect wet weight of small intestinal mucosa,the height and width of microvilli,malonaldehyde(MDA)level and myeloperoxidase(MPO)activity.At the same time,mesenteric lymph node,liver and splenic tissue were taken to perform bacterial culture.Bacterial translocation was observed.RESULTS:After supplement,72 rats were involved in the result analysis.①Plasm FITC-dextran concentration of IR group were higher than that in control group,DIPC group and RIPC group,respectively(P＜0.01).②Plasm xylose concentration in the IR group was lower than that in the control group,DIPC group and RIPC group,respectively(P＜0.01).③Bacterial translocation rate in the IR group was higher than that in the control group,DIPC group and RIPC group,respectively(P＜0.01).④Small intestinal mucosal injury degree in the IR group was lower than that in the other 3 groups(P＜0.01).⑤Small intestinal MPO activity and MDA level in IR group were significantly higher than those in the other 3 groups(P＜0.01). Serum SOD activity and NO level were lower but amylase activity and TNF-α 1evel were higher in the IR group as compared with the other 3 groups(P＜0.01).CONCLUSION:IPC of two hindlimbs in both donor and recipient can protect small intestinal mucosal barrier and reduce bacterial translocation rate after pancreas transplantation in rats.

BACKGROUND: Pancreas transplantation alone (PTA) is an effective therapy for diabetic patients who do not occur chronic complications. It's important to establish the stable PTA animal models to investigate immunologic tolerance or ischemic/reperfusion injury.OBJECTIVE: To establish the model of pancreas transplantation alone (PTA) with enteric drainage in rat.DESIGN: Grouping and controlled animal experiment SETTING: Department of Gastrointestinal Surgery, Xijing Hospital,Fourth Military Medical University of Chinese PLA MATERIALS: Totally 90 SD male rats, with the body mass of 250-320 g,were chosen in this study. 58 rats were induced by intravenous administration of streptozotocin (STZ) at a dose of 65 mg/kg via penile vein and the rats whose fasting plasma glucose exceeded 19.4 mmol/L for more than 2weeks were selected, 22 rats was successful. Rats were randomly assigned to 2 groups: control group (10 healthy rats) and group PTA consisted of 22diabetic rats, which received PTA from 22 normal donors.METHODS: This experiment was conducted at the laboratory of Department of Gastrointestinal Surgery, Xijing Hospital, Fourth Military Medical University of Chinese PLA from January 1999 to July 2004. The blood vessels reconstruction of PTA were performed using end-to-side anastomosis between the donors' abdominal aorta segment (abdominal artery and splenic artery) and recipients' abdominal aorta, and end-to-end anastomosis between the donors' portal vein segment (splenic vein) and recipients'left renal vein (use a cuff). Pancreas exocrine drainage was made by pancreas intestine anastomosis (Roux-Y).MAIN OUTCOME MEASURES: Body mass, food intake, water intake and fasting blood glucose were monitored 2 days before operation and 1,3,7,14 and 30 days after operation, and the failure causes were analyzed.RESULTS: 22 rats in the model group and 10 rats in the normal control the vein of the rats , very severe diabetic symptoms appeared in 22 rats:Body mass, food intake, fasting blood glucose was increased than that of cipients operation was (32.2±12.7) minutes and (63.4±15.9) minutes respectively. And the mean time of warm and cold ischemic time was 0minute and (48.6±18.3) minutes, respectively. 11 of the 22 cases (50%)died or lost their function of the endocrine within 1 month in Group PTA.The main complications were secondary pancreatitis and pancreas leakage after transplantation (7 cases, 31.8%). All successful recipients' blood glucose lowed on the 1st and recovered to be normal on the 3nd after transplantation (P ＜ 0.01), and their food intake, water intake and urine volume decreased and became stable 14 days later.CONCLUSION: This method can be used to establish relative stable animal model. Successful PTAs may improve the pancreatic endocrine function of the diabetic rats.

Objective Thymic epithelial tumors(TET) consist of thymoma and thymic carcinoma. The purpose of this study was to investigate the diagnostic value of 18 F-fluorodeoxyglucose (FDG) coinci-dence SPECT imaging in TET. Methods 18 F-FDG coincidence SPECT images of 37 TET patients ( male 16, female 21, age range 24-82 years; mean age 55 years) were retrospectively analyzed according to the simplified WHO histological classification: high survival thymoma (types A, AB and B1 ), median survival thymoma (types B2 and B3 ), and low survival thymoma (thymic carcinoma). Qualitative visual interpreta-tion was based on foci of18F-FDG uptake'higher than that of normal mediastinum. Turnour/lung radioactivity ratio (TLR) was calculated from the counts of region of interest (ROI) drawn around the mass and over a comparable area of normal lung tissue in TET patients. The distribution of18F-FDG uptake was compared with the enhanced CT findings. TLR was also correlated with the Ki67-1abeling index. Statistical comparison for TLR in different groups were performed with variance analysis. Results Thirty-four TET were positive and 3 negative ( A, A, AB) by 18 F-FDG coincidence SPECT images. The differences among the 3 sub-groups were statistically significant (F=9.99,P<0.05). The TLR of these 3 subgroups was variable: 1.42±0.27, 2.13±0.74, 3.00±1.19, respectively. A close correlation (r=0.613, P=0.002) of the TLR with the Ki67 labeling index (n= 22) was found. In addition, 18 F-FDG coincidence SPECT detected two metastases ( anterior chest wall and right supraclavicular fossa lymph node) that were not identified on enhanced CT. Conclusions 18 F-FDG coincidence SPECT is useful in differentiating subgroups of thymic epithelial tumors and in identifying more invasive thymic lesions.18 F-FDG may be a novel tracer that ena-bles in vivo evaluation of cellular proliferation in human TET.

OBJECTIVE:To compare the effects of repeated subcutaneous insulin administration vs. insulin pump continuous subcutaneous administration on related indexes of advanced age patients with gestational diabetes mellitus(GDM). METHODS:Inretrospective study,120 advanced age patients with GDM were randomly divided into group A(60 cases)and group B(60 cases). Group A was given Insulin aspart injection with initial dose of 0.5 U/(kg·d)subcutaneously before meal,adjusted according to fast-ing blood glucose(FPG)and postprandial 2 h blood glucose(2 hPG),and then given Isophane protamine biosynthetic human insu-lin injection with initial dose of 0.5 U/(kg·d)subcutaneously at bedtime,adjusted according to FPG and 2 hPG. Group B was giv-en Insulin aspart injection with initial dose of 0.5 U/(kg·d)added into insulin pump using 40% of total daily insulin as basic pump,increasing to 60% of total daily insulin if blood glucose control was poor,adjusted according to FPG and 2 hPG. Treatment course of 2 groups lasted for 4 weeks. The time of blood glucose reaching target,the amount of insulin were observed in 2 groups, and the levels of FPG,2 hPG,HbA1c,Hcy and Cys-C before and after treatment,the occurrence of patients and neonates compli-cations. RESULTS:The time of blood glucose reaching standard in group B was significantly shorter than group A;the amount of insulin,the incidence of hypoglycemia,premature birth,excessive amniotic fluid and gestational hypertension in group B were sig-nificantly lower than group A,with statistical significance(P<0.05). The incidence of neonatal respiratory distress syndrome in group B was significantly lower than group A,with statistical significance(P<0.05);but there was no statistical significance in the incidence of giant baby and malformation between 2 groups(P>0.05). After treatment,the levels of FPG,2 hPG,HbA1c,Hcy and Cys-C in 2 groups were significantly lower than before treatment,and the group B was significantly lower than the group A, with statistical significance(P<0.05). CONCLUSIONS:The insulin pump continuous subcutaneous administration is significantly better than repeated subcutaneous insulin administration in respects of controlling glucose level,reducing the amount of insulin,the levels of Cys-C and Hcy,maternal and neonatal complications.

Pancreas transplantation alone(PTA) which dose not popularize in clinical therapy the insulin-dependent diabete mellitus patients who have not occurred end-stage complications,which was resulted in not only the technical but also the sociological and ethnics reasons.There are some reasons to restrict the development of clinical PTA at present.In this article the sociological and ethnics reasons to circumscribe the development of clinical PTA were discussed.

Objective:To explore blood type distribution of newborns hemolytic disease ( HDN ) caused by maternal and neonatal blood type incompatibility and analyze the value of hemolysis three trials in the diagnosis of HDN.Methods:Hemolysis three trials of type O or Rh negative maternal cord blood samples and hyperbilirubinemia of the newborn blood samples from January 2014 to 2016 were detected by micro-column gel test cards.Then the results were statistically analyzed.Results:(1) There were 918 cases of maternal and neonatal blood type incompatibility in all 1350 cases.569 cases were detected HDN positive with the rate of 62%( 569/918).Among 569 cases,the positive rate of direct anti-globulin test,free antibody test and antibody released test were 27.9%(159/569),86.5%(492/569) and 100% respectively.There was statistical difference of the combination of direct anti-globulin test negative,free antibody test positive and antibody released test positive compared with other combinations ( P<0.05 ).( 2 ) There was statistical difference of HDN positive rate between ABO 73.8%(551/747) and Rh 10.5%(18/171)in 918 cases of blood type incom-patibility.(3)There was statistical difference between A positive rate of 80%(280/350) and B positive rate of 68.3%(271/397) in 747 cases of ABO incompatibility.(4)There was statistical difference among RhD positive rate of 17.7%(14/79),RhE positive rate of 6.8%(4/59) and RhC positive rate of 0(0/33).Conclusion: Antibody released test was the most sensitive test in hemolysis three trials to diagnose HDN.The probability of HDN positive caused by maternal and neonatal ABO blood type incompatibility was significantly higher than Rh.The probability of HDN positive with type A newborns was significantly higher than type B.The probability of HDN positive caused by RhD blood type incompatibility was significantly higher than RhE and RhC.