Objective To study the method of obtaining a large number of dendritic cells （DC）. To study the specific cytotoxicity T lymphocyte (CTL) effect against tumor cells initiated by DC pulsed with peptide of cancer cell. Methods Development of cells with cytologic features of DC in bone marrow cultures supplemented with granulocyte macrophage-colony stimulus factor (GM-CSF) and IL-4. Determining the DC phenotype and the specific structure by electronic microscopy. The CTL effect against pancreatic carcinoma leading by the DC pulsed with tumor cells lysate in vitro was observed. Results A large number of typical DC was proliferated by supplementing with GM-CSF and IL-4 cytokines. DC had specific cell appearance and structure, and highly expressed various cell surface molecules. TNF-α had the ability of stimulating DC mature, the mature DC had the enhancing abilities of antigen presenting and IL-12 self-secreting, as well as, expressed higher levels of CD54， MHC-Ⅱ and CD86 molecules than control group (P＜0.05). T lymphoid cell stimulated by DC without tumor antigen could not recognize and kill the target cells, only if DC pulsed with peptide of cancer cell can lead a b immune response to special tumor cells. The inhibiting ratio of CTL was significantly higher than that in other groups (P＜0.01). Conclusion Bone marrow DC has b ability of inducing special CTL against determined cancer cells after they are pulsed with tumor cell lysate. DC vaccine is probably a new immunotherapeutic method against tumor in the near future.
【Abstract】ObjectiveTo study the positive effect of recombinant human epidermal growth factor (rhEGF) on rabbit intestinal anastomotic wound healing after bowel resection. MethodsFortyeight white rabbits were randomly divided into study group in which rhEGF was injected and spinged in the submucosa and mucosa respectively during intestinal anastomosis after bowel resection, and control group in which only intestinal resection and anastomosis was performed. The leukocyte was counted. The incidence of anastomotic leakage and the synthesis of collagen fibrils and hydroxyproline were observed. ResultsThe leukocyte numbers in the anastomotic tissue in two groups rabbits increased slightly 3 d, 5 d and 7d after intestinal anastomosis, but the difference between study group and control group was insignificant (Pgt;0.05). The incidence of anastomotic leakage in the control group (16.7%) was higher than that of the study group (4.3%). The area of collagen fibrils 3 d, 5 d and 7d after intestinal anastomosis in the study group were significantly more than that in the control group (P＜0.05). Number of fibroblast was higher in the study group and the cells appeared bigger nucleus and dense colouration as well as enriched plasm. Angiogenesis in anastomosis tissue in the study group was significant and normal structure was present. Cell structure of anastomosis mucosa was damaged in the control group. Synthesis of hydroxyproline in anastomotic tissue 5 d and 7 d after anastomosis in the study group was more than that in the control group (P＜0.05).ConclusionInflammation was present in the whole process of wound healing, and local using of EGF had insignificant effect on system inflammation. EGF functions as chemoattractant and increases the recruitment of leukocytes, monocytes and fibroblasts into the wound area. EGF increases the production of collagen, angiogenesis and the synthesis of hydroxyproline. So EGF could promote wound healing and protect from anastomosis leakage in this study.
【Abstract】ObjectiveTo explore the effect of glutamine on immune function of rat with obstructive jaundice and its possible mechanism. MethodsFifty male Wistar rats were randomly divided into three groups: Control group (n=10)， obstructive jaundice group (n=20) and glutamine treatment group (n=20). The serum concentration of TNF-α, IL-10 was detected by using radioimmune method. Liver function was measured through automated biochemistry analyzer. The animal model of obstructive jaundice was established by ligating the rat’s common bile duct. Bacteria cultures were performed with the rat’s tissues of lung, spleen, liver and kidney respectively. ResultsCompared with control group, obstructive jaundice group showed statistically lower serum level of TNF-α, and statistically higher serum level of IL-10, TBIL, ALT and AST during the first and the second week after ligation of common bile duct. During the first and second week after administration of glutamine, the serum TNF-α of glutamine treatment group was statistically higher than that in control group and obstructive jaundice group. Meanwhile, glutamine treatment group showed statistically lower serum level of IL-10, TBIL, ALT and AST than obstructive jaundice group. There were statistically less bacteria translocations in glutamine treatment group than those in obstructive jaundice group. Conclusion Glutamine can increase the immune function by changing serum concentration of TNF-α, IL-10 and decrease the bacteria translocation.
目的探讨脾动脉瘤的诊治经验。方法对我院1980～2000年收治的2例脾动脉瘤患者的临床资料进行回顾性分析。结果例1为男性，因脾功能亢进入院，行CT及血管造影检查发现脾动脉多发动脉瘤，行动脉瘤切除、脾切除术。例2为女性，因腹腔内出血而急诊行多普勒检查得以确诊，急诊手术行远端脾动脉结扎、脾切除术。结论对临床症状明显且有增大趋势的脾动脉瘤应积极行手术治疗，特别是预期妊娠的女性患者及行肝移植患者。妊娠患者，推荐介入及腹腔镜微创治疗方法。
The conectration of cholecystokinin infasting serum was determined by radioimmunoessay in 30 patients with gastric antrum cancer before and after radical sbutotal gastrectomy.It was 119.6±142.2pmol/L before the operation and 78.5±149.2pmol/L after the operation,which was significantly lower than that before the operation,P=0.022. The result suggests that the reduction of cholecytokinin secretion after gastrectomy was one of the important causes in the bile stasis,the disturbance of gallbladder emptying funcion and the formation of gallstone.
【Abstract】Objective To investigate the expressions of TNF-α and superoxide dismutase (SOD) mRNA in myocardium of rats with obstructive jaundice (OJ). Methods The expressions of TNF-α and SOD mRNA were semi-quantitatively analyzed after amplification of cDNA in myocardium of the rats with OJ by RTPCR. Results The expression of TNF-α mRNA increased and that of SOD mRNA decreased in the myocardium of rats with OJ. The level of plasm TNF-α increased and SOD synthesis in myocardium decreased. Conclusion The injury of myocardium in OJ is correlated with increasing expression of TNF-α mRNA and decreasing expression of SOD mRNA.
ObjectiveTo evaluate the effect of ZnPP Ⅸ on the expressions of heme oxygenase-1 (HO-1) and glutathione-Stransferase-π （GST-π） and the chemosensitivity of drug-resistant hepatic carcinoma cell line Bel/Fu, and explore it’s possibility to reverse drug-resistance and the relevant regulating mechanism. Methods①MTT assay was adopted to detect the drug sensitivity for adriamycin, mitomycin, and 5-fluorouracil of Bel/Fu cell after ZnPP Ⅸ being induced for 24 h. ②RTPCR was carried out to detect the expressions of HO-1 and GST-π mRNA after Bel/FU cells being treated with different concentrations ZnPP Ⅸ for 24 h. ResultsAfter Bel/Fu cells being treated with ZnPP Ⅸ for 24 h, the 50% inhibiting concentration (IC50) for drugs was decreased dramatically （Plt;0.05）. Meanwhile, the expressions of HO-1 and GST-π mRNA in the treated cells also decreased dose-dependently (Plt;0.01). ConclusionsZnPP Ⅸ can increase the chemosensitivity of Bel/FU cells by down-regulation of HO-1 and GST-π expression. ZnPP Ⅸ is a potential agent to reverse multidrug resistance of hepatic carcinoma cells.
Objective To explore the effect of dendritic cells (DCs) allergized by K-ras mutant peptide on expressions of chemokines CCL19, CCL22, and cytoskeletal protein fascin-1. Methods DCs were derived from peripheral blood in the presence of granuloceyte/macrophage-colony stimulating factor, interleukin (IL) -4 in vitro. The DCs were collected on day 7 after culture, and were divided into non-K-ras mutant peptide group (addition of RPMI 1604 culture solution 50 μg/ml) and K-ras mutant peptide group (addition of K-ras mutant peptide 50 μg/ml). Phenotype was identified by flow cytometry. The morphological structure was observed by scanning and transmission electron microscopies, respectively. The expressions of IL-12, CCL19, and CCL22 were tested continuously by enzyme-linked immunosorbent assay (ELISA). The expression of cytoskeletal protein fascin-1 was determined by Western blot. Results ①The expressions of CD1a, CD80, and CD86 after loading K-ras mutant peptide were higher than that before loading K-ras mutant peptide (Plt;0.01). ②The DCs with petal-like and branch-like profections after loading were observed under scanning electron microscopy; The DCs with irregular shapes, branch-like or burr-like were showed under transmission electron microscopy. ③The expressions of IL-12, CCL19, and CCL22 in the Kras mutant peptide group were higher than those in the non-K-ras mutant peptide group at different times (6, 12, 24, and 48 h) after loading Kras mutant peptide (Plt;0.01). ④The expression of fascin-1 in the K-ras mutant peptide group was also higher than that in the non-K-ras mutant peptide group (Plt;0.01). Conclusion K-ras mutant peptide can promote DC to mature and improve the expression of chemokines and cytoskeletal protein which will strengthen DC migration.
Objective To explore the effect and mechanism of glutamine to the aberrant crypt foci (ACF) in rat injured by acetic acid. Methods Thirty Wistar rats were averagely divided into three groups: control group, acetic acid group and glutamine group. The colon of the rat was infused with 1% acetic acid. Started to gavage with glutamate two days after modeling glutamine group. The injured colons were studied after fourteen days with light and scanning electronic microscope. Paraffin sections of specimens were prepared and stained with HE. The colon crypts were isolated by HCl digestion method. The expressions of CD44 and ICAM-1 in the epithelial cell of the large intestine mucosa were detected by immunohistochemistry method. Results On the days of 14， the number of ACF in the glutamine group were remarkably decreased as compared with that of the acetic acid group and a branch-like. The expressions of ICAM-1 and CD44 (every 1 000 cells) were 302.1±30.1 and 298.6±28.3 in glutamine group, 223.6±23.5 and 221.5±28.6 in control group, 198.5±19.5 and 215.3±17.8 in acetic acid group, respectively. While the expressions of CD44 and ICAM-1 in intestine were increased remarkably in the glutamine group compared with the control group and acetic acid group (P＜0.05). Conclusion Glutamine could decrease the formation of the ACF injured by acetic acid. Increasing the expressions of CD44 and ICAM-1 may be one of the important factors to decrease the ACF.
Objective To investigate the effect of renal cell apoptosis induced by obstructive jaundice on the expression of bcl-2 in rats, and to explore the mechanism of renal impairment induced by obstructive jaundice. Methods Thirty-two male SD rats were randomly divided into 2 groups: SO group and BDL group. The rats in SO group received sham operation. Bile ducts of rats in BDL group were ligated. Pathology of kidneys was observed under the microscope. The levels of D-Bil, TBA, GOT, GPT, Cr and BUN in serum and β2-MG in urine were measured. The apoptotic rate of renal cells was calculated by flow cytometry and the forms of DNA fragmentation in renal cells were detected by agarose gel electrophoresis. The expression of inhibitory gene bcl-2 in the renal tissues was detected by immunohistochemistry. Results The color of urine in BDL group became dark yellow in day 2 after operation; The ears, tails and the muscle of abdominal wall and splanchnic organs, such as liver and kidney, also became yellow and swollen in day 7. The D-Bil, TBA, GOT, GPT, BUN of serum and β2 -MG of urine in BDL group were higher than those in SO group (P＜0.05, P＜0.01), and each value (except β2 -MG) in BDL group of 14 d was higher than that in BDL group of 7 d (P＜0.05, P＜0.01), respectively. The result of flow cytometry showed that the apoptotic rate of SO group and BDL (7 d and 14 d) group were (2.10±0.75)%， (18.17±0.86)% and (36.39±2.23)% respectively, there were significantly difference among them (P＜0.05). The expression rate of bcl-2 of renal cell in BDL group of 7 d was higher than that in BDL group of 14 d. Conclusion Obstructive jaundice could induce apoptosis of the renal cells, and activate the expression of bcl-2 of the renal tubular epithelial cells in feedback, which may regulate the process of apoptosis.