Objective To investigate the short-term and long-term effect of cornary-caval shunt accompanied by pericardial devascularization in the treatment of upper gastrointestinal bleeding caused by portal hypertension.Methods Eleven patients with portal hypertension underwent cornary-caval shunt accompanied by partial pericardial devascularization were chosen.Of the 11 patients 6 applied autogenous splenic veins for graft and in 5 cases the coronary vein and inferior vena cava were anastomosed directly.Results Of the 11 patients,no operative mortality or early rebleeding.All patients were followed up from 5 months to 11 years with an average of 5 years and 3 months,of whom two died,others having no rebleeding or hepatic encephalopathy.Conclusion Cornary-caval shunt is a highly selective portosystemic shunt.Cornary-caval shunt accompanied by pericardial devascularization is a surgical treatment of upper gastrointestinal bleeding caused by portal hypertension for its apparent regional antihypertensive effect,the normal blood flow of liver,and reduction of the incidence of rebleeding.

Objective To observe effects of ginsenoside Rb1 on phosphorylation of microtubule-associated protein 2 (pMAP-2) in hippocampus and amygdala of depressive model rats. Methods Thirty male Wistar rats were randomly divided into control group, model group and treatment group. The depression rat model was produced by giving chronic unpredicted mild stress (CUMS). Treatment group was given daily intragastric administration of ginsenoside RB 1 (1 g/mL crude drug, 1 mL/100 g body weight) for 22 days during modeling. Western blot assay was used to detect expressions of MAP-2 and pMAP-2 protein, and real-time PCR was used to detect expressions of pMAP-2 mRNA respectively. Results The expressions of pMAP-2 protein and mRNA in hippocampus and amygdala were significantly lower in model group than those of control group (P < 0.05). The expressions of pMAP-2 protein and mRNA were significantly higher in treatment group than those of model group (P<0.05). Conclusion Ginsenoside Rb1 can play anti-depression role by inhibiting the phosphorylation of MAP-2 in rats.

Objective:To compare the histological differences between magnetic anastomosis and traditional suture in canine portal vein (PV) .Methods:Eighteen healthy Chinese garden dogs, either gender, 8-12 months and weighing 13.5-18.9 kg, were randomly divided into magnetic compressive anastomats (MCA) group ( n=9) and hand-sewing (HS) group ( n=9) for PV reconstruction. The time of PV anastomosis was compared between the two groups. HE and Masson staining were performed immediately and at 4, 8, 12 and 24 weeks after operation. The ultrastructure of the anastomosis was observed using scanning and transmission electron microscopy. Results:All dogs survived. The PV anastomosis time was significantly shorter in MCA group (3.58±2.75) min than that HS group (12.89±3.12) min, P<0.01. In MCA group, the vascular wall of anastomotic stoma was well aligned immediately after operation, and the shrinkage was obvious in HS group by gross eyes. At 24 weeks, electron microscope scanning showed the re-endothelialization was smooth and endothelial cells arranged regularly at the anastomotic site of the MCA group, whereas different-sized and irregularly aligned endothelial cells and large collagenous fibers arranged in disorder were present at the HS anastomotic stoma. Representative HE and Masson staining confirmed that the magnetic device was associated with decreased infiltration of inflammatory cells and deposition of fibrotic collagen at 24 weeks explanted anastomotic stomas compared with the HS group. Conclusions:Compared with the HS, MCA produced shorter anastomosis time, smooth anastomotic intima, light fibrous tissue hyperplasia, no foreign body residue, mild inflammatory reaction and reliable technique for canines PV anastomosis.

Aim To investigate the effects of endogenous and exogenous hydrogen sulfide(H_2S)on the proliferation and apoptosis of human fetal lung fibroblasts.Methods Human fetal lung fibroblasts were cultured under 2% O2～93% N_2～5% CO_2 for 24 h to produce hypoxia.Cells were divided into 6 groups:(1)Hypoxia group(N_2);(2)N_2+600 μmol·L~(-1) NaHS group;(3)N_2+1 200 μmol·L~(-1) NaHS group;(4)N_2+6 400 μmol·L~(-1) NaHS group;(5)N_2+400 μmol·L~(-1) cysteine(Cys)group;(6)N_2+200 μmol·L~(-1) S-adenosyl-L-methionine(SAM)group.After they were cultured for 24 h, MTT assay was used to evaluate the cell proliferation, and flow cytometry was used to detect cell apoptosis.Results Compared with N_2 group, 600 and 1 200 μmol·L ~(-1) NaHS(H_2S donor)significantly reduced proliferation induced by hypoxia of human fetal lung fibroblasts(P <0.01)without effects on apoptotic rates of cells(P >0.05)and 6 400 μmol·L~(-1) NaHS increased apoptosis of human fetal lung fibroblasts during hypoxia significantly(P <0.05), although no effects were found on proliferation of cells(P >0.05).In addition, Cys, substrate of cystathionine β-synthetase(H2S synthase, CBS) or SAM(activator of CBS)did not affect proliferation of human fetal lung fibroblasts induced by hypoxia(P >0.05), whereas apoptotic rates were increased significantly compared with that of N_2 group(P <0.05).Conclusions Endogenous and exogenous hydrogen sulfide can inhibit proliferation induced by hypoxia and promote apoptosis of human fetal lung fibroblasts, suggesting endogenous hydrogen sulfide may play a protective role through lung fibroblasts by inhibiting the pulmonary vascular structural remodeling caused by hypoxia.

Objective To study the effect of gender on the curative effect of rituximab for the treatment of diffuse large B cell lymphoma with five-year progression-free survival. Methods The clinical and laboratorial data of 155 diffuse large B cell lymphoma patients from January 2003 to January 2011 were retrospectively analyzed. The patients were divided it into R-CHOP group and CHOP group according to if rituximab was used, and then subdivided based on their gender into R-CHOP-M, R-CHOP-F groups and CHOP-M, CHOP-F groups, respectively. Kaplan-Meier survival curve was plotted for the progression-free survival. Results The five-year progression-free survival rate in the R-CHOP group was higher than the CHOP group, and the rate of R-CHOP-F group was higher than R-CHOP-M group, but there were no significant differences between the CHOP-M group and the CHOP-F group. The rate of the R-CHOP-M group was a little bit higher than the CHOP-M group with no statistical significance. The rate of the R-CHOP-F group was higher than CHOP-F group. Conclusion Rituximab is beneficial for female than for male, which may contribute to the adjustment of drug doses if a male is treated with rituximab.

Objective To explore the feasibihty of TRAIL to be used to remove the leukemia cells from the autologous hemopoietic stem cell transplants. Methods The expression of decoy receptor 1 and decoy receptor 2 on the bone marrow mononuclear cell were routine-ly isolated and observed by fluorescence microscope after PE-DcR1 or PE-DcR2 stain. The apeptosis rates of mononuclearcell and Jurkat cells interfered by 200ng/ml TRAIL for 18h were determined by flow cytometry after AnnexinV/Pl stain. The interfered mononuclear cells were cultured to count the number to form colony-forming unit-fibroblast(CFU-F). The Jurkat cells labeled by green fluorescent protein were in-corporated into the mononuclear ceils and affected by 200ng/ml TRAIL for 24h. The incorporated cells were cultured in the system with GM-CSF and EPO and the numbers of the CFU-GM, BFU-E and fluorescence colony were counted on the seventh day. Results Both decoy re-ceptor land decoy receptor 2 of TRAIL can be detected on membrane or in cytoplasm of the bone marrow mononuclear cell. The apoptosis rate of mononuclear cell interfered by 200ng/ml TRAIL for 18h was (5.95±1.23)%, which was markedly lower than that of Jurkat cells (33.42±2.28) %. The number of CFU-F of TRAIL group and control group were 235.67 ~ 33.56 and 249.33±42.72, respectively. No marked difference can be found between the mentioned two groups. Moreover, TRAIL decreased the number of fluorescence colony formed by Jurkat cells without significant decreasing the number of CFU-GM and BFU-E formed by bone marrow mononuclear cells. Conclusion TRAIL can selectively induce apoptosis in Jurkat cells without marked toxic effect on the bone marrow mononuclear cells, which means that TRAIL can be used to remove the leukemia cells from the autologous hemopeietic stem cell transplants in vitro.

Objective To investigate the chemotherapeaties sensitivity and expression of bcl-2, bcl-xL and bax of Jurkat cells co-cultured with bone marrow stromal cells (BMSC) isolated and cultured from leukemia patients. Methods BMSC were isolated and cultured from leukemia patients routinely. To construct the co-cultured model, Jurkat cells were co-cultured with of the irradiated layer BMSC by 60Co and observed the model with scanning electron microscope. The Jurkat cells suspension-cultured were used as control. The apoptosis and IC50 were detected by the FACS can machine and MTT, respectively. The expression of bcl-2,bcl-xL and bax in Jurkat cells was detected by Western blotting. Results We found that the Jurkat cells in the model showed a decreased sensitivity to DNR, IC50 values for leukemic BMSC and nonadhered contol were of 2.30 μmol/L and 0.45 μmol/L, respectively. Moreover, Jurkat cells adhered to BMSC have a survival advantage over suspended cells following DNR exposure for 24 h, apoptosis percentages for leukemic BMSC group and nonadhered controls were of (6.05±0.54)% and (25.74±6.15)%, respectively. As compared with controls, leukemic BMSC group had significant difference in apoptosis percentages (P <0.01). The expression of bcl-2 in Jurkat cells was up-regnlated when adhered to BMSC for 4 h and the higher expression emerged after adhering for 24 h and 48 h. No marked change of bcl-xL and bax expressions were observed in the adhered Jurkat cells. Conclusion The adhered-culture with bone marrow stromal cells isolated from leukemia patients could make the leukemia cells acquire drug resistance, which was associated with the up-regulated expression of bcl-2 in the leukemia cells.

Objective To investigate the effects of adhesion mediated by bone nlalTow stroma celh from leukemia patient on chemotherapeutics sensitivity and cell cycle of Jurkat cells in the co-cultured model.Methods Bone mw stroma cells were isohted and cultured from leukemia patients routinely.To construct the co-cultured model.Jurkat cells were co-cultured with BMSCs the irradiated layer by 60Co,and the model was observed with scanning electron microscope.The IC50 values of Jurkat cells expesured to DNR were quantified by MTT.The cell cycles of Jurkat cells after 24h-adhesion in the co-cultured model were analyzed by Facs.The expression of cyclin A,cyclin E and p27 in Jurkat cells adhered to BMSCs for 4h.24h and 48h were detected by Western blot.Results Jurkat ceUs in the co-cultured model showed a decreased sensitivity to DNR.ICSO values for normal BMSCs,leukemic BMSCs and non-adhered control were of 1.78Ixmol/L,2.30pznol/L and 0.45p,mol/L,respectively.The percentages of Go-Gl phase for leukemic BMSCs group and non-adhered control group were of 48.74％±8.77％and 27.83％壬1.86％.Respectively.The percentages of Gz-M phase for leukemic BMSCs group and non-adhered control group were of2.01％±1.17％and 20.33％±1.84％。Respectively.Compared with eomrol group,the 24h-ad- hesion mediated by BMSCs from leukemia patients up-regulated the percentage of Go-G1 phase of Jurkat cells(P

BACKGROUND:Leukemia cells can obtain drug resistance phenotype mediated by adhesion to bone marrow stromal cells. But, for chronic myelogenous leukemia with adhesion functional defects, the role and mechanism of bone marrow stromal cells in imatinib-resistant formation remain unclear. OBJECTIVE:To construct the co-cultured model of bone marrow stromal cells-K562 cells and to investigate the influences of the co-culture with bone marrow stromal cells from the patients with chronic myelogenous leukemia on imatinib sensitivity of K562 cells and cellcycle. METHODS:The co-culture model was constructed by co-culturing K562 cells with bone marrow stromal cells isolated and cultured from the patients with chronic myelogenous leukemia. The IC50 values of K562 cells exposed to imatinib were quantified by MTT assay. The apoptotic rates of K562 cells exposed to 0.5μmol/L imatinib for 72 hours were detected by flow cytometry through Annexin V-FIT/PI labeling. The cellcycles, cellcycle protein (cyclin A, cyclin D1 and cyclin E) expression of K562 cells co-cultured with bone marrow stromal cells for 72 hours were analyzed by flow cytometry.RESULTS AND CONCLUSION:The IC50 values of co-culture group and suspension culture group were respectively (0.52±0.02)μmol/L and (1.27±0.05)μmol/L, and their comparison showed significant differences (P<0.01). After 72 hours of treatment with 0.5μmol/L imatinib, the apoptotic rates in the co-culture group and suspension culture group were respectively (15.48±4.17)%and (32.01±6.83)%, and their comparison showed significant differences (P<0.01). The percentages of G0-G1 phase of K562 cells co-cultured with bone marrow stromal cells for 72 hours were (48.81±8.27)%, which were significantly higher than the suspension culture group (25.78±3.26%) (P<0.01). The co-culture with bone marrow stromal cells from the patients with chronic myelogenous leukemia could mediate K562 cells resistance to imatinib. The mechanism was possibly related with G0/G1 arrest of K562 cells induced by co-culture with bone marrow stromal cells.

BACKGROUND:CTLA-4Ig as a tolerance-induction agent is a potential strategy in graft-versus-host disease prevention. OBJECTIVE:To investigate the efficacy of CTLA4Ig-gene-modified bone marrow stromal cels mediated by adenovirus to induce T-cel tolerance of haploidentical donors. METHODS: The bone marrow stromal cels isolated culture from the bone marrow of HLA haploidentical donors were transfected by recombinant adenovirus encoding CTLA4IgcDNA (AdCTLA4Ig) at a multiplicity of infection=50 for 72 hours. The expression rate and the location of CTLA4Ig in the transfected cels were detected by fluorescence microscope after immunofluorescence staining. CTLA4Ig-modified bone marrow stromal cels (2×104, 4×104and 8×104) were respectively co-cultured with 105 T cels from the peripheral blood of HLA haploidentical donors and 105 peripheral blood mononuclear cels from recipients. The proliferative inhibition rate was determined by MTT assay, and the level of interleukin-2 in the supernatant was detected by ELISA. The bone marrow mononuclear cels (1×105/wel) were co-cultured with CTLA4Ig-modified bone marrow stromal cel layers constructed in 6-wel plates. The number of bone marrow mononuclear cels and colony-forming unit-granulocyte macrophages were calculated after 5-day culture. RESULTS AND CONCLUSION: The expression rate of CTLA4Ig at the multiplicity of infection=50 was as high as 85%, and the immunofluorescence signals of CTLA4Ig were distributed unevenly in the cytoplasm. The inhibition rates of 2×104, 4×104, and 8×104 CTLA4Ig-modified bone marrow stromal cels on proliferation of T cels were higher than that of untransfected cels. The levels of interluekin-2 in the corresponding cel groups were significantly lower than that in the untransfected cels (P < 0.05). At 5 days of culture, there was no significant difference in the number of bone marrow mononuclear cels and colony-forming unit-granulocyte macrophages between the transfected and untransfected cel groups (P > 0.05). These findings indicate that CTLA4Ig-modified bone marrow stromal cels mediated by adenovirus can induce immune tolerance of T-lymphocyte from HLA haploidentical donors in vitro.