Objective TO compare the effect of early postoperative intraperitoneal chemotherapy (EPIC) with that of intravenous chemotherapy (IVC)on the immunity of patients with gastrointestinal carcinoma (GC).Methods 36 patients were divided into two groups each with 18 cases, one group received EPIC and the other IVC, then determined the serum T-lymphocyte subsets (T LS) and sIL-2R before and after the chemotherapy.Results Serum CD3, CD4 and sIL 2R in GC patients after the chemotherapy decreased more significantly than those of before the chemotherapy (P0.1),but mean sIL 2R level in EPIC group was lower than that in IVC (0.05

Objective To study and summarize the cause and reoperative method of massive hemorrhage of upper gastrointestinal tract after gastrectomy. Methods The clinical data of 11 cases patients with massive hemorrhage of the upper gastrointestinal tract after gastrectomy from August 1986 to June 2000 were analyzed retrospectively. Results 5 cases were anastomotic bleeding,3 cases injured gastric remnant mucosa bleeding,2 cases ulcer bleeding after bancroft gastrectomy, 1 case overlooked leiomyoma bleeding.All patients were cured with reoperation.Conclusions To prevent post operative bleeding is the key. The evaluation of the condition of patients with massive hemorrhage of the upper gastrointestinal tract,timing of reoperative intervention,and the selection of proper operative method are also very important.

Objective To investigate the effects and mechanism of Safflower Injection on the proliferation,apoptosis,and expression of bcl-2/bax gene in hepatic stellate cell(HSC) in vitro.Methods HSC Line HSC-T6 was incubated with Safflower Injection at different concertration,cell proliferation was assessed by MTT colorimetric assay.After incubated with Safflower Injection 5,10,and 20 mg/mL,flow cytometry(FCM) was used to detect the content of DNA in HSC-T6.Morphological change of HSC-T6 was observed under microscope and agarose gel electrophoresis for DNA Ladder was used to detect apoptosis.Besides,the early stage of apoptosis was detected with Annexin V-FITC/PI double labbled assay.And real time RT-PCR was used to detect the expression of bcl-2/bax gene.Results The significant inhibition of Safflower Injection on HSC proliferation was observed in a dose-and time-dependent manner.Observed by FCM,the cell ratio in G0/G1 phase with Safflower Injection treatment(10 and 20 mg/mL) for 24 h was increased,which showed the significant difference compared with the control group(P

Objective To investigate the effect of bone marrow derived endothelial progenitor cells (EPC)transplantation on microenvironments in a murine model of chronic vein thrombosis.EPCs transplantation was evaluated whether it can up-regulate thrombus organization and recanalization associated cytokines(VEGF,ANG-1 and MCP-1). Method EPCs from immature Wister rats' bone marrow were isolated using a Ficoll density gradient centrifugation,and cultured in fibronectin-coated plate in EGM-2M Vmedium.EPCs were harvested on the 10th day,then were transplanted into chronic inferior vens cava thrombus of adult Wister rat through the femoral vein.Rats were divided into three groups:blank control group(group A,sham operation),the control group(group B,the medium injected)and the experimental group(group C,EPCs injected).The rats were sacrificed after 28 days.VEGF,ANG-1 and MCP-1 mRNA was measured by real-time quantitative PCR and protein expression change by Western blotting from IVC and thrombus tissue. Results EPCs were identificated successfullv by immunohistochemistry,immunofluorescence and function,then were transplanted into chronic inferior vena cava thrombus of adult rats.After EPCs transplantation,the VEGF,ANG-1 and MCP-1 mRNA expression in group C expression was significantly up-regulated with statistical significance(P＜0.01)compared with group A and group B in IVC and thrombus tissue by real-time PCR.There was no significant difference between group A and group B (P＞0.05).VEGF,ANG-1 and MCP-1 protein expression were similar to mRNA expression.There was significant increase in group C compared to group A and group B(P＜0.01)and no statistical significance between group A and group B(P＞0.05).Conclusion EPCs deriving from bone marrow may change the microenvimnment of chronic vein thrombus through up-regulating thrombus organization and recanalization associated cytokines(VEGF,ANG-1 and MCP-1).

Objective To investigate the role of Hedgehog pathway in hepatic fibrosis and its association with activation of hepatic stellate cells. Methods Twenty male Spragur-Dawley rats were divided into control and model groups with 10 each. The animal models were induced by injection with CCl4 and fed with fat-rich diet. The rats in both groups were sacrified at the 8 week with 5 each and the liver tissues were removed for HSC-T6 culture. The deposition of collagen fiber in liver was detected with HE and Masson staining. RT-PCR was used to detect the expressions of Sonic hedgehog (Shh), smoothened (Smo), patched (Ptc), Gli-1 and α-smooth muscle action (α-SMA) mRNA in HSC-T6 and liver tissues. The influence of cyclopamine (Cyc) and lipopolysaccharide (LPS) on HSC-T6 proliferation were assayed by MTT. The expressions of Shh, Smo, Ptc, Gli-1 and α-SMA mRNA after intervention with Cyc (100μmol/L) and LPS were measured by real-time PCR. Results A lot of lipo and collagen deposited in liver of model rats. The Shh,Smo,Gli-1 and α-SMA mRNA were highly expressed in model rats than those in control group (2-△△Ct were 20.45±3.31 vs. 1, 12.78 ± 0. 53 vs. 1, 10.88 ± 2.41 vs. 1, 4.91 ± 2. 59 vs. 1, respectively, all P value <0. 05). In vitro Cyc inhibited HSC-T6 proliferation in dose dependant manner (F=636.81, P<0.01). Compared to the control group, the mRNA expressions of Smo, Ptc, Gli-1,α-SMA in HSC-T6 were significantly reduced after Cyc intervention (2△△Ct, were 0. 20±0. 11, 0. 21 ± 0. 08, 0. 28 ± 0. 05,0. 27±0.10,respectively, all P values<0.01). Conclusion The expression of members of Hedgehog pathway are increased in the progress of hepatic fibrosis, which may accelerate the hepatiee fibrosis by activating HSC.

Objective To study the effect of endothelial progenitor cells(EPCs) transplantation on chronic deep venous thrombosis.Methods Bone marrow-derived mouonuclear cells (BMMNCs) were isolated from rat bone marrow by ficoll and cultured with EGM-2MV medium.A rat model of chronic deep vein thrombosis was established by partial ligation of the inferior vena cava and intravenous injection of thrombosin.Model rats were randomly divided into three groups:A(n =25),EPCs group,1 ml 10~6 EPCs transplantation;B(n = 25),EGM-2MV medium group,1 ml EGM-2MV medium transplantation;C (n =25),control group,without any treatment.After transplantation,HE staining and immunohistochemical staining was conducted to detect recanalization of the inferior vena cava.Western blotting of inferior vena cava thrombosis was used to detect VEGF,bFGF protein expression changes.SPSS13.0 software was used for analysis.Results Compared with group B and C,VEGF,bFGF protein significantly increased in group A.The recanalization capillary density was significantly higher in group A than that in group B,and C (P ＜0.05).The neovascularization was identified by immunohistochemical staining using vWF antibody,as endothelial cells.Conclusions EPCs were the precursor of endothelial cells,when transplanted into the deep vein thrombos,initiating angiogenesis and accelerating organization and recanalization of vein thrombus.

Objective To analyze expression of interleukin (IL)-23p19 and IL-23 receptor (IL-23R) in inflammatory bowel disease (IBD),and the role in the induction of peripheral blood T cell activation and proinflammatory cytokine secretion.Methods Peripheral blood (PB) and intestinal mueosal biopsies were collected from 12 patients with Crohn's disease (CD),25 patients with ulcerative colitis (UC) and 20 healthy controls.Expression of IL-23p19 was determined by immunohistochemistry and RT-PCR.IL-23R expression in CD4+,CD8+ T and NK cells from peripheral blood and lamina propria was analyzed by flow eytometry.Peripheral blood mononuclear ceils (PBMC) were isolated and cultured under stimulation with IL-23 and anti-CD3,and the levels of tumor necrosis factor α (TNFα) interferon (IFN)γ and IL-2 were determined by enzyme-linked immunosorbent assay (ELISA).Results The expression of II.-23p19 was significantly increased in inflamed mucosa of CD at both the transcriptional and translational levels compared with that in UC and healthy controls.IL-23R was mainly expressed in PB- and lamina propria-CD4+,CD8+T cells and NK cells from IBD patients,and markedly increased compared with controls (P＜0.05).IL-23 strongly triggered PBMC from IBD patients to produce significantly higher levels of IFN-γ,TNF-α and IL-2(P＜0.05).Conclusions IL-23p19 and IL-23R are highly expressed in IBD,particularly in CD,and may play an important role in the induction of T cell activation and proinflammatory cytokine secretion,suggesting that targeted therapy directed against IL-23 may have a therapeutic role in IBD.

Objective To investigate the role of pancreatic β cell on pancreatic regeneration following experimental acute pancreatitis.Methods Eighty-seven SD male rats were randomly divided into four groups:control group ( n =15 ),STZ group ( n =24),L-Arg group ( n =24 ),STZ + Arg group ( n =24).60 mg/kg of STZ was administrated by intraperitoneal injection to induce the diabetes model.2.5 g/kg body weight of LArg was administrated by intraperitoneal injection to induce the acute pancreatitis model.The rats were sacrificed 1,3,5,7 d later and the serum levels of amylase and glucose were measured.Relative pancreatic weight (pancreatic weight/body weight) were measured.Pancreatic tissue underwent routine pathologic examination,and the percentage of area of necrosis and tissue transformation was calculated.The expression of Reg4 and insulin was performed by immunofluorescence.Results Serum level of glucose significantly increased after STZ injection.After L-Arg injection,serum level of amylase significantly increased,and there was pancreatic tissue edema,necrosis,infiltration of inflammatory cells,which suggested the successful model induction.The percentage of area of necrosis in STZ + L-Arg group was (71.6 ± 6.0) ％ at the 3rd day,which were significantly higher than (42.3 ± 4.0 ) ％ in L-Arg group; the percentage of area of transformation was (45.6 ± 5.4) ％,which were significantly lower than (78.5 ± 6.4) ％ in L-Arg group.Expression of Reg4 in pancreatic islets of STZ + L-Arg group was significantly lower than those in L-Arg group.Conclusions STZ impairs pancreatic β cells,aggravates pancreatic damage following L-arginine induced pancreatitis and inhibits pancreatic regeneration.

Objective To investigate the expression of Iroquois homeobox protein 1 ( IRX1 ) gene and the hypermethylation status of its promoter in pancreatic cancer,and their relationship.Methods Real-time PCR was used to quantitatively detect IRX1 gene expression level of 12 sets of resected pancreatic cancer tissue and 6 sets of pancreatic cancer cell lines; gene sequences analysis was used to detect the structure of IRX1 promoter; DNA methylation inhibitor 5-Aza-2′-deoxycytidine (5-Aza-dC) was used in pancreatic cancer cell lines,and then the methylation of IRX 1 was measured by methylation-specific PCR (MSP) and unmethylation sequence-PCR (USP) methods.Results Expression of IRX1 mRNA in pancreatic cancer tissue was 0.31 ± 0.11,which were significantly lower than that in normal pancreatic tissue ( 1.05 ±0.32,P ＜0.01 ).IRX1 mRNA expression of AsPCl,BxPC3,Capan 2,PANCl,PaTu8988 and SW1990 were 0.36 ± 0.08,0.34 ±0.16,0.37 ±0.11,0.25 ±0.06,0.31 ±0.04,0.36 ±0.02,which were significantly lower than that in human kidney epithelial 293 cells ( 1.03 ± 0.28,P ＜ 0.05 or ＜ 0.01 ).Analysis of IRX1 gene sequence showed abundant CpG islands in promoter.Hypermethylation of IRX1 promoter site was found in all pancreatic cancer cell lines.However,its methylation status could be reversed by 5-Aza-dC,and the IRX1 expression was also restored.Conclusions IRX1 mRNA expression is down-regulated in pancreatic cancer,and it is related with promoter CpG islands hypermethylation.

In this study antigen-independent factor in the pathogenesis of chronic rejection of organ transplants was examined. Kidney isografts and allografts were transplanted orthotopically into bilaterally nephroectomized rat recipients and studied functionally, morphologically and immunohistologically, at serial intervals up to 52 weeks after transplantation. Allograft recipients developed progressive proteinuria after 12 weeks, with gradual renal failure ultimately leading to death. At the same time, morphological changes, including progressive arteriosclerosis and glomerulosclerosis, tubular atrophy and interstitial fibrosis, developed. Immunohistologically, macrophages infiltrated glomeruli during this period and cytokines became unregulated. Our results showed that antigen-independent functional and morphological changes occurred in long-term kidney isografts and mimicked those appearing much earlier in allografts that reject chronically. Initial injury and extent of functioning renal mass is suggested to be important factor for such late changes.
Animals ; Graft Rejection ; etiology ; immunology ; pathology ; Graft Survival ; physiology ; Kidney ; immunology ; pathology ; Kidney Transplantation ; immunology ; methods ; pathology ; Proteinuria ; etiology ; Rats ; Rats, Inbred Strains ; Time Factors ; Transplantation, Homologous ; Transplantation, Isogeneic