Objective To identify energy metabolism-proteins of mice midbrain by using the proteomic technique,and to investi-gate the relationship between these proteins and neural diseases.Methods Two-dimensional gel electrophoresis was used to sepa-rate totally soluble proteins extracted from mice midbrain.Some protein spots on two-dimensional gel electrophoresis gels were ana-lyzed by using matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).Results 24 protein spots related with energy metabolism were separated by two-dimensional gel electrophoresis and identified by using MALDI-TOF MS successfully.Conclusion The establishment of energy metabolism-related proteins map of mice midbrain lays a foundation for the research on the involvement of these proteins in neural disease pathogenesis.

Objective To investigate the genetic heterogeneity of 5’-NCR of HCV genotype 1b.Methods We selected 147 HBV genotype 1b serum samples. HCV 5’-NCR fragments were amplified from these samples by use of RT-PCR assay and sequenced after using restriction endonuclease Mbo Ⅰ and BamH Ⅰ. We analyzed the phylogenetic trees of the samples and compared them with 40 isolates of HCV genotype 1b.Results The sequencing reports indicated the isolates of HCV genotype 1b recognized by BamHⅠat position 117 by a substitution (C-T) at position 120 in HCV genotype 1b viruses in China. Of 147 samples, 17 (11.56%) samples for one BamHⅠrecognization position, 26 (17.69%) for neither BamH Ⅰ recognization position nor Mbo Ⅰ,6 samples (4.08%) for two Mbo I recognization position, 18(12.24%)for one MboⅠrecognization position.Conclusions Of 147 HBV genotype 1b serum samples, 54.42% for one Mbo I recognization position, whether this have relationship with the response to IFN therapy provide the framwork for future detailed investigation of HCV antviral therapy. There is specific BamH I recognization position in isolates of HCV genotype 1b in China as compared to the other 40 HCV 1b isolates. The role of this specific mutation needs to be further researched.

Objectives In our studies before, we reported mutation C-T at -222 of hepatitis C virus type 1b genotyped on 5′ noncoding region (5′NCR). There was no this mutation of type 1b recorded in GenBank. Since genotypiog based on 5′NCR cannot differentiate subtypes, it is necessary to investigate whether the hepatitis C virus genotype 1b strains with mutation C-T belongs to other subtypes of genotype 1. Methods 64 HCV genotype 1b samples were amplified from 5′NCR by RT-PCR.Then the products were digested by use of BamHⅠ restriction enzymes. We randomly chose 6 samples with BamHⅠ restriction site and subjected them to amplification of 5′NCR and NS5B. The sequences of 5′NCR were analyzed. Sequences of NS5B fragments from the 6 samples were respectively subjected to phylogenetic analysis with subtypes of genotype 1 and 38 complete genomes of genotype 1b from GenBank.Results The phylogenetic analysis of 6 samples and subtypes of genotype 1 indicated that the genotypes of 6 strains with BamHⅠ restriction site in 5′NCR were type 1b, instead of subtype of type 1. Tree construction with 38 complete genomes of genotype 1b showed that the 6 mutants of genotype 1b did not belong to the same branch of the tree and there were no genetic differences between the mutants and the other strains of genotype 1b.Conclusions Our research indicated that the hepatitis C virus stains of C-T in 5′NCR were mutants of type 1b. Since the 5′NCR is a highly conserved region of HCV, the mutation might have some relationship to the long-time IFN therapy.

Objective To study the clinical effect of peritoneal and abdominal indwelling tube by ultrasound for patients with severe acute pancreatitis.Methods Sixty-eight patients with severe acute pancreatitis from January 2010 to December 2013 were analyzed retrospectively.The patients were divided into experiment group (48 cases) and control group (20 cases).The patients in control group were given the common drugs treatment,and the patients in experiment group were given the common drugs and the peritoneal and abdominal indwelling tube by ultrasound.The clinical effects and complication were compared.Results The total efficacy rate in experiment group was 93.75％(45/48),in control group was 35.00％(7/20),and there was significant difference (P ＜ 0.05).Seven patients in experiment group occurred complication,after the symptomatic treatment 6 patients recovered and 1 patient died of multiple organ failure;7 patients in control group occurred complication,after the symptomatic treatment 3 patients recovered and 4 patitents died of infection recurrence.The complication rate in experiment group was 14.58％(7/48),in control group was 35.00％ (7/20),and there was significant difference (P ＜ 0.05).Conclusion Peritoneal and abdominal indwelling tube by ultrasound could improve clinical efficacy and decrease complication rate for patients with severe acute pancreatitis.

Objective To evaluate catheterization in pancreatic duct before endoscopic papillary bal-loon dilation (EPBD)to prevent pancreatitis after EPBD.Methods Forty-three patients with normal serum amylase levels,diagnosed as having bile duct stones,underwent EPBD.Twenty-three were assigned to experi-mental group,where catheters(ERCP imaging tube)were placed in pancreatic duct before EPBD,then the pa-pillary balloon was expanded to 10 mm.Twenty were assigned to control group where eight-millimeter-diameter papillary balloon was used to remove the stones.The serum amylase levels before EPBD,6 hours and 24 hours after EPBD,the incidence of pancreatitis and high serum amylase levels associated with EPBD,as well as the mean time and success rate of removing the stones of the two groups were compared.Results Post-EPBD pan-creatitis occurred in one patient in experimental group (4.35%),and seven in control group (35.00%), which was significantly different(P <0.05).Meanwhile,the mean levels of serum amylase 6 h and 24 h after EPBD in the experimental group were (102.61 ±98.99)U /L and (60.35 ±26.18)U /L respectively,lower than those in the control group (398.25 ±259.32)U /L and (230.50 ±281.31)U /L(P <0.05).After the papillary balloon was expanded to 10 mm in experimental group,the mean time of removing stones was (10.43 ±2.27)min,which was shorter than that of control group (17.90 ±4.49)min (P <0.05).Stone-re-moving rate of two groups had no difference and they all succeeded one time.Conclusion Placing catheter in pancreatic duct before EPBD to prevent pancreatitis after EPBD makes it easier to remove stones in shorter op-eration time.It can prevent pancreatitis and high amylase blood disease after EPBD.

Objective To explore the curative effect of triple lumen feeding tube on upper gastroin-testinal tract perforation after endoscopic operation.Methods A total of 1 12 patients with upper gastrointes-tinal perforation after endoscopic operation were involved in this prospective control test.The remission rate of abdominal pain and abdominal distension,gastric pH,lesion healing time,length of stay and the success rate of conservative treatment were compared between the control group (indwelling traditional gastric tube and intravenous use of omeprazole injection)and the experimental group (intestinal administration of PPI through triple lumen feeding tube).Results The remission rate of abdominal pain of the experimental group was 61. 3% at 6 hours,and 83. 2% at 12 h,which were significantly higher than those of the control group (P<0. 05 ).The lesion healing time(of cases where lesions were not clipped)and length of hospitalization of the experimental group were significantly shorter than those of the control group(P<0. 05 ).The gastric pH and success rate of conservative treatment of the two groups had no significant difference(P>0. 05 ).Con-clusion The curative effect of triple lumen feeding tube on upper gastrointestinal tract perforation after endo-scopic operation is ideal.

Objective To investigate the anti-tumor activity of triterpenoids from Ganoderma lucidum on different tumor cells . Methods The MTT assay was adopted to detect the in vitro inhibition effect on 5 kinds of tumor cells .The inhibiting curve was drawn ,IC50 was calculated for reflecting the compound′s cytotoxic activity .Results The in vitro experiments demonstrated that three kinds of triterpenoids compound monomer showed different degrees of inhibition effect ,in which the inhibitory effect of gano-derenic acid Y was stronger ,its IC50 on H460 lung cancer cells was 22 .4 μmol/L ,followed by 7-oxo-ganoderic acid Z2 ,its IC50 was 43 .1 μmol/L .Conclusion Ganoderenic acid Y shows a strong inhibitory activity on H 460 lung cancer cells ,7-oxo-ganoderic acid Z2 shows a certain inhibitory activity on H 460 lung cancer cells ,moreover the inhibitory activity is dose dependent .The three com-pounds of ganoderenic acid Y ,7-oxo-ganoderic acid Z2 and ganoderon B have no activity or very weak activity to the other detected cell lines .The anti-lung cancer activity of ganoderenic acid Y and 7-oxo-ganoderic acid Z2 needs to be further deeply studied .

BACKGROUND:Adipose-derived stem cells are easily col ected and abundantly cultured, which can proliferate rapidly when being cultured in vitro. With multi-directional differentiation potential, adipose-derived stem cells are expected as seed cells for tissue engineering.
OBJECTIVE:To isolate, culture and identify of adipose-derived stem cells from Sprague-Dawley rats in vitro.
METHODS:The subcutaneous adipose tissue was obtained from the iliac region of rats under the aseptic condition, and then was digested with 0.075%type Ⅰ col agenase and cultured in vitro. The morphology and proliferation characteristics of the cells were observed under an inverted phase contrast microscope. The third passage was put into gauge for growth curve by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, and the cells were also identified by CD44, a stem cellmarker, with immunofluorescence staining. Adipose-derived stem cells were induced and differentiated into adipocytes in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10%fetal bovine serum, dexamethasone and insulin, and then the cells were identified with oil red“O”staining. Adipose-derived stem cells were induced and differentiated into neural cells, and then the cells were identified with immunohistochemical staining.
RESULTS AND CONCLUSION:The growth curve of adipose-derived stem cells was opposite-like“S”shape, and it strongly expressed CD44 that can designate a stem cell. The passage cells were exposed to a defined medium for adipocyte differentiation, and then could be stained with oil red. After being induced and differentiated into nerve cells, the cells expressed neuron-specific enolase. The adipose-derived stem cells of Sprague-Dawley rats are characterized by easy isolation, culture and proliferation in vitro, expressing related phenotypes of mesenchymal stem cells, as wel as induction and differentiation under certain conditions.

Objective To evaluate endoscopically insertion of feeding tube via nose(EIFTN). Method Jejunal feeding tube was placed endoscopically via the nose in all 136 coma patients. Results This procedure was successful in all patients. The procedure took an average time of 5 minutes. In patients with deep coma,the procedure had no influence on HR,MAP, ECG and SaO2; In semicoma patients, HR and R increased during the procedure (t=3.902, P

Objective To observe the effect of costimulatory molecule B7-1 on cytoskeleton rearrangement in mouse podocytes induced by angiotensin Ⅱ (Ang Ⅱ),and to study the underlying molecular mechanism of B7-1 in the pathological changes of podocytes.Methods All cultivation of conditionally immortalized mouse podocytes (MPC) in vitro were divided into the following groups:normal control group,CTLA-4 group,Ang Ⅱ group (10-6 mmol/L 12 h,24 h;10-8 mmol/L 12 h,24 h) and CTLA-4 with Ang Ⅱ group.Transfect B7-1 RNA interference fragment (siRNA) to the mature podocytes,and then restimulated by Ang Ⅱ (10-6 mmol/L 12 h),the change of podocyte cytoskeleton after Ang Ⅱ stimulation were observed.The expression of B7-1 in each group was assayed by flow cytometry and Western blotting.The nephrin and p-nephrin protein levels in the four groups were also analyzed by Western blotting.At the same time,the podocyte cytoskeleton distribution as indicated by F-actin was observed by fluorescence microscopy.Results Flow cytometry and Western blotting showed that B7-1 was not expressed in the normal control group.Ang Ⅱ showed a concentration and time dependent induction of B7-1 expression in mouse podocytes (P ＜ 0.05).Western blotting indicated that Ang Ⅱ induced B7-1 protein expression (P ＜ 0.05).Expression of nephrin and p-nephrin was significantly down-regulated by Ang Ⅱ (P ＜ 0.05).Compared with the normal control group,the expression of podocyte protein nephrin and p-nephrin in Ang Ⅱ stimulation group was significantly reduced (P ＜ 0.05).Using FITC phalloidin fluorescence staining showed that CTLA-4+Ang Ⅱ stimulation group cytoskeleton rearrangement was improved significantly and F-actin recombinant score (mCFS) decreased compared with Ang Ⅱ group (P ＜ 0.05),suggesting that Ang Ⅱ led to the disorder of the podocytes cytoskeleton and the destruction of the cytoskeleton of podocytes by Ang Ⅱ could be improved after B7-1 blocking.Compared with the Ang Ⅱ stimulation,transfection of B7-1 siRNA + Ang Ⅱ stimulation group improved F-actin cytoskeletal rearrangement,and mCFS also decreased significantly (P ＜ 0.05),suggesting that transfection of B7-1 siRNA might improve the damage of Ang Ⅱ on podocytes cytoskeleton.Conclusions B7-1 participates in the process of cytoskeleton reconstruction and plays an important role in the pathological changes of podocytes.