Humans ; Severe Combined Immunodeficiency ; therapy

Objective To investigate causes and treatment approaches for postoperative complications after laparoscopic adjustable gastric banding(LAGB).Methods Clinical and follow-up data of 302 cases were reviewed.The body mass index (BMI),percent excess weight loss (％ EWL),operation time,intraoperative blood loss,the incidence of complications and management were analyzed and summarized.Results There were no conversion to open surgery.The overall complication rate was 6.29％ including 2 cases of gastric wall injury,5 cases of gastric banding slippage (recovered by reoperation).There was no gastric parietal banding corrosion,tube bursting leakage,pulmonary embolism,micronutrient deficiencies,nor mortality.Conclusions The majority of patients were satisfied with the operation effect,still,there were substantial postoperative complications including gastric wall injury,gastric banding slippage.

Objective To investigate the method for effective establishment of nude rat tumor xenograft model of human lung cancer cells A549 in order to provide the experimental basis for tumor-related interventional research in vivo. Methods A549 cell lines were subcutaneously transplanted in nude rats, then single-cell suspension or tumor tissue block were prepared when the tumor lesion was established. The single-cell suspension and tumor tissue block were transplanted into subcutaneous tissue behind ear in rats. The tumor formation rate, growth situation and cell cycle of primary xenograft tumor group, the secondary single-cell suspension group and the secondary tumor block group were evaluated. The results were analyzed. Results The tumor formation rate of the secondary tumor block group was significantly higher than that of the other two groups. The tumor cells quickly proliferated with less tumor variation. Tumor cell cycle analysis indicated that G2/M ratio of the secondary tumor block group was remarkably higher than that of the other two groups. Conclusion Transplantation with tumor tissue block can significantly increase the tumor formation rate of human lung cancer cells A549 in experimental rats. This technique is an effective method for the establishment of nude rat tumor xenograft model.

Age impact in mouse model of secondary hepatic alveolar echinococcus was investigated in this research . Twenty-nine 8-week-old ,twenty-five 18-week-old and twenty-five 28-week-old female mice were anesthetized with 20% ure-thane by intraperitoneal injection and then transhepatically injected by Echinococcus multilocularis (E .m) tissue suspension through skin incision and abdominal muscle to liver in all three groups to establish mouse model of secondary hepatic alveolar e-chinococcus .Results showed that the survival rates for the three groups of mice were 62 .1% ,84% and 68% ,respectively (P>0 .05) .The E .m infection rates in liver were 72 .2% ,71 .4% and 76 .5% ,respectively (P>0 .05) .The diameter of E .m cysts in liver were 0 .915 ± 0 .103 cm ,1 .247 ± 0 .112 cm and 1 .215 ± 0 .197 cm ,respectively (P>0 .05) .The mass of E .m cysts in liver were 0 .332 ± 0 .035 g ,0 .532 ± 0 .155 g and 0 .382 ± 0 .085 g ,respectively (P> 0 .05) .HE stain showed no difference in pathology .Results indicated that the establishment of secondary hepatic alveolar echinococcus model by using transhepatic injection through skin incision and abdominal muscle of 18-week-old mice was capable of simplifying operation and improving the survival rate of the mice .

Objective To analyze predictive factors on survival in patients with completely resected high-risk Ⅱ/Ⅲ stage colorectal cancer after adjuvant chemotherapy.Methods According to random number table, 76 cases with completely resected high-risk Ⅱ/Ⅲ stage colorectal cancer after adjuvant chemotherapy were selected, who newly diagnosed and hospitalized in 2004. Their disease-free and overall survivals were followed up.Thymidylate synthase gene polymorphism and microsatellite instability were tested in these cases with microdissection combined with polymerase chain reaction and capillary electrophoresis. Correlation of these factors including clinical characteristics, thymidylate synthase gene polymorphism and microsatellite instability to survival was analyzed with SPSS13.0 software. Results Histologic grades and evaluated lymph node number had significantly difference between two groups of distinct prognosis (χ2 = 7.827, P =0.003 and χ2 = 9.265, P =0.018, respectively), which were also independent predictors on survival proved by COX regression analysis (χ2 = 40.472, P =0.000 and χ2 = 39.528, P =0.000, respectively).Kaplan-Meier survival analysis showed that the median disease-free and overall survival of poor-differentiated adenocarcinoma patients were significantly shorter than those of high and intermediate-differentiated ones (27.67 vs 61.13months, χ2 = 45.015, P =0.000 and 43.13 vs 64.21 months, χ2 = 35.514, P =0.000, respectively), as well, the median disease-free and overall survival of patients with the evaluated lymph node number less than 11 were poorer than those of more than 11 ( 45.65 vs 68.47 months, χ2 = 23.134, P =0.011 and 53.10 vs 70.18months, χ2 = 22.896, P =0.013, respectively).Conclusion Poor-differentiated adenocarcinoma and evaluated lymph node number less than 11 may be predictors on poor survival in patients with completely resected highrisk Ⅱ/Ⅲ stage colorectal cancer after adjuvant chemotherapy.

Objective To clone mcp1, mcp2 and mop3 genes that encoding methyl-accepting chemotaxis proteins(MCP) of Campy/obacter jejuni for construction of their prokaryotic expression systems, and to establish chemotactic model in vitro of the microbe for determining chemotaxis-inducing substances and to understand relationship among MCPs and chemotactic inducers. Methods The segments of mep1, mcp2 and mcp3 genes were amplified by PCR and then sequenced after T-A cloning. Prokaryofic expression systems of the genes were subsequently constructed. SDS-PAGE pins Bin-Rod Gel Image Analyzer were used to examine the expression of target recombinant proteins rMCP1, rMCP2 and rMCP3, and Ni-NTA affinity chromatography was performed to purify the rMCPs. Rabbits were immunized with each of the three rMCPs to obtain antisera. Immunodiffusion assay was performed to measure the titers of antisera. IgG in each of the antisera were extracted by ammonium sulfate precipitation and DEAE-32 ion exchange chromatography, and IgG F(ab')2 were then prepared by pepsin enzymolysis and Sephadex G-100 chromatography. Chemotactic model in vitro of C. jejuni based on HAP( hard-agar plus) method was established to determine the chemo-taxis-inducing effect of eight candidate substances. Chemotaxis inhibition test based on IgG F(ab')2 bloc-king was applied to determine the function and diversity of MCPs. Results The segments with expected si-zes amplified from mcp1, racp2 and mcp3 genes were obtained by PCRs, and their nucleotide and putative amino acid sequences were completely same as the reported. The constructed prokaryotic systems could effi-ciently express rMCPs with the yields about 10% of the total bacterial proteins. Immunization with rMCP1, MCP2 and rMCP3 enables the rabbits to produce specific antibody. All the antisera had 1: 4 immunodiffusion titers. Both bovine bile and sodium deoxycholate(DOC) were able to induce ehemotactie movement of C. je-juni in a dosage-dependent manner ( P < 0.05 ). When MCP1 and MCP2 were blocked with their IgG F(ab')2, the ehemotaetic ability of C. jejurd were remarkably decreased (P <0. 05). However, MCP3 blocking did not affect the chemotaxis ( P > 0.05 ). Conclusion The C. jejuni MCPs are successfully ex-pressed in this study. Bovine bile and DOC are the inducers for chemotaxis of C. jejuni, and MCP1 and MCP2 are involved in the process of ehemotaxis iadueed by DOC.

Objective To compare the inhibiting effect on human lung adenocarcinoma who were treated with gem?citabine combined with cisplatin chemotherapy through either arterial or intravenous route to explore the optimum adminis?tration route. Methods Human lung adenocarcinoma derived A549 cells were transplanted into 40 BALB/c-nu mice to es?tablish lung cancer model. The models were divided into 4 groups:animals in arterial or intravenous chemotherapy groups were treated with gemcitabine 150 mg/kg combined with cisplatin 10 mg/kg through either arterial route or intravenous route. Animals in negative control group were given normal saline through caudal vein while animals in sham operation group were treated with normal saline via arterial route. Then dynamical change of tumor volume and tumor inhibiting rate were assessed , and Bcl-2 and Caspase 3 expressions were investigated using western blot. Finally inhibiting effect were compared between these two different administration routes. Results Transplanted tumors in arterial and intravenous che?motherapy groups (especially in arterial group) were suppressed, in terms of mass of tumor(g:1.91±0.19, 2.61±0.21 vs 4.58± 0.46), compared to the control group (P<0.05). Furthermore, tumor inhibiting rates in arterial chemotherapy group and ve?nous chemotherapy group are 57.6%and 42.4%respectively (P<0.05). Expression of Bcl-2 was down regulated while ex?pression of Caspase-3 was up regulated upon both arterial and intravenous chemotherapy. And arterial route showed much more obvious tumor apoptosis effect than venous route. Conclusion Arterial route of gemcitabine combined with cisplatin for lung adenocarcinoma treatment is more effective to restrain the tumor growth in clinical application.

In order to observe the effects of different culture media and temperature on protoscoleces of Echinococcus multi‐locularis ,they were randomly divided into RPMI‐1640 group ,D‐MEM group and M199 group ,and cultured in three degrees of temperature (4 ,25 and 37 ℃) with 10% fetal calf serum (FCS) .Protoscoleces were counted by light microscope with 0 .1%eosin staining ,and calculated survival rate (per 100 protoscoleces) everyday until all the parasites died .At the same time ,the average number of the preservation days was observed .The experiment results showed that the survival rate of protoscoleces in RPMI‐1640 and D‐MEM groups were higher than that in M199 group (P<0 .05) and there’s no significant difference between RPMI‐1640 group and D‐MEM group (P>0 .05) .The survival rate of protoscoleces in RPMI‐1640 group at 4 ℃ and 37 ℃and D‐MEM group at 25 ℃ were higher ,but there was no significant effect of 4 ,25 and 37 ℃ on the survival rate of proto‐scoleces (P>0 .05) .Significant difference were found in the survival rate of protoscoleces on the 3rd day and the 9th day in these three groups (P<0 .05) .The average number of the preservation days were 34 days in RPMI‐1640 group at 4 ℃ ,36 days in D‐MEM group at 25 ℃ and 23 days in M199 group at 4 ℃ .It was concluded that the effects of different culture media and tem‐perature on protoscoleces are different ,and the RPMI‐1640 at 4 ℃ and D‐MEM at 25 ℃ are more suitable for culturing proto‐scoleces in v itro .

Objective To investigate the function of phosphatidylinositol phospholipase C encoded by LB361 gene of L.interrogans ( L-PI-PLC) and its mechanism in inducing macrophage apoptosis .Meth-ods The PI-PLC domains in the sequence of LB 361 gene of L.interrogans serovar Lai strain were analyzed by bioinformatics method .Prokaryotic expression system was established to express the recombinant L -PI-PLC ( rL-PI -PLC).The enzymatic activity of rL-PI-PLC in hydrolyzing phosphatidylinositol -4,5-bisphos -phate (PIP2) substrate into inositol-1,4,5-trisphosphate (IP3) was determined by IP3 fluorescence polariza-tion assay.LB361gene expressions at mRNA and protein levels as well as the secretion of LB 361gene prod -ucts were detected by real-time fluorescent quantitative RT -PCR and Western blot assay after infection of hu-man THP-1 macrophages with L.interrogans serovar Lai strain.A LB361 gene-transfected THP -1cell line was generated for evaluation of the mechanism of LB 361 gene products in elevating intracellular free Ca 2+( [Ca 2+] i) concentration and inducing the apoptosis of transfected THP -1 cells with the use of laser confocal microscopy and flow cytometry.Re sul ts The rL-PI-PLC hydrolyzed PIP2 into IP3 with a Km of 199 μmol/L and a Kcat of 8.566×10-5 S-1 .The expressions of LB361gene at mRNA and protein levels were both signifi -cantly up-regulated after infection of THP-1 cells with L.interrogans serovar Lai strain .Moreover , the exter-nal secretion of L-PI-PLC was also found during infection .The concentrations of IP 3 and [ Ca2+] i in the LB361 gene-transfected THP-1 cells were significantly increased compared to those in the non-transfected THP-1 cells, resulting in a high [Ca2+]i-dependent apoptosis of partial THP-1 cells.Conclusion PI-PLC is encoded by the LB361 gene of L.interrogans, which could induce the apoptosis of macrophages through el-evating [ Ca2+] i concentration during infection of microphages with L.interrogans.

Objective To explore the prognostic significance of p53 mutation protein in patients with diffuse large B-cell lymphoma for the purpose of individualized therapy. Methods Newly diagnosed 62 cases were randomly chosen from our hospital, p53 mutation protein and CD10, bcl-6, MUM1 were tested by immunohistochemistry. Correlation of p53 mutation protein with patients ' characteristics, genotype and survival were analysed in the study. Results p53 mutation protein was found in 48.4 % (30/62) of patients.Its expression was only related to initial treatment response (x2 =20.365, P =0.040), including complete remission rate of 33.3 % (10/30) in positive group and 59.4 % (19/32) in negative group, and non-germinal center genotype (x2=31.023, P =0.021) with 83.3 % in positive group and 56.2 % in negative group. No other correlation was not verified with clinical features. Multivariate survival analysis showed that p53 mutation protein was an independent predictor for shorter progress-free and overall survival in positive group (x2 =30.784, P =0.005 and x2 =35.276, P =0.006). Conclusion p53 mutation protein should be an independent predictor with poor prognosis and to direct personalized therapy.