1.Effects of glutamine enriched parenteral nutrition on mucosal ischemia and reperfusion injury of small bowel in rats
Lipan PENG ; Leping LI ; Changqing JING
Chinese Journal of Current Advances in General Surgery 2009;0(10):-
Objective:Small bowel IRI models in rats were established to explore the effect of glutamine enriched parenteral nutrition on mucosal barrier,and to discuss the probable mechanisms.Methods:Thirty Wistar rats were randomly assigned into 3 groups:The control group(N group,n=10) ,conducted fictitious operation and fed with common forage,TPN group(n=10) and TPN+Glu group(n=10) .The morphous of mucous,serum and intestinal mucosal Gln concentration,levels of D-lactate,endotoxin,TNF-?,IL-6,HO-1 positive ratio and HO-1 mRNA were detected.Results:Glutamine obviously improved the structure of intestinal mucosal and decreased the expressions of D-lactate,endotoxin,TNF-?and IL-6.And enhanced the expressions of HO-1 mRNA and HO-1.Conclusion:Glutamine enriched parenteral nutrition can alleviate small intestinal IRI and inflammatory reaction and enhance the HO-1 and HO-1 mRNA expressions.HO-1 and its metabolin's anti-oxygen,anti-apoptosis,anti-inflammator action may be the mechanism of the protective action of Gln on mucosal barrier of small bowel.
2.Reversal of multidrug resistance in human colon carcinoma LOVO/5-Fu cells by tetrandrine
Kailei WANG ; Leping LI ; Changqing JING
Chinese Journal of General Surgery 2011;26(5):367-370
Objective To explore the reversal effect on MDR1 gene-mediated multidrug resistance in human colon carcinoma LOVO/5-Fu cells by tetrandrine ( Tet) and to clarify its molecular mechanism.Methods LOVO/5-Fu cells were treated for 48 h with Tet.Drug sensitivity was measured by MTT.The cell cycle, apoptosis of cells and expression of P-glycoprotein (P-gp) were determined by flow cytometry assay.Expression of MDR1 mRNA was detected by real-time quantitative PCR (real-time PCR).P-gp expression was detected by Western blot.Results After LOVO/5-Fu cells were treated for 48 h with Tet, the IC50 of 5-Fu decreased to ( 4.15 ± 0.31 ) μg/ml ( P < 0.05 ) ; and the apoptotic rate increased to (3.44% ± 0.28% ) ( P < 0.05) ; the expression of MDR1 mRNA reduced to (570 ± 85) (P < 0.05 ).Conclusions Tetrandrine reverses MDR1 gene-mediated multidrug resistance in human colon carcinoma LOVO/5-Fu cells possibly by inhibiting the expression of MDR1, decreasing the expression of P-gp, thus enhancing the sensitivity of LOVO/5-Fu cells to 5-fluorouracil.
3.Application of multi-slice computed tomography in the diagnosis of colorectal neoplasms
Ximing WANG ; Yanhua DUAN ; Leping LI
Chinese Journal of Digestive Surgery 2010;09(6):465-467
Colorectal neoplasms always present with thickness of the intestinal wall or a soft tissue mass in the enteric cavity. Multi-slice computed tomography (MSCT) with high spatial resolution and advanced post-processing techniques can demonstrate the above signs of the tumor, and the invasive signs of adjacent structures and lymph node metastasis. Combined with three dimensional reformation images, MSCT shows a higher sensitivity than that of double contrast barium enema and electronic colonoscope. MSCT is promising in the diagnosis and preoperative evaluation of colorectal neoplasms.
4.Expression of VEZT in gastric cancer tissues and the construction of VEZT over-expression vector
Yansen LI ; Xiaobo GUO ; Changqing JING ; Huiliang ZHANG ; Leping LI
Chinese Journal of General Surgery 2014;29(10):767-770
Objective To investigate the expression of VEZT and its clinical pathological significance in gastric cancer tissues,and to construct the VEZT over-expression vector.Methods The expression of VEZT was examined in 119 cases of gastric carcinoma and their corresponding normal mucus tissues by SP immunohistochemical staining.The relationships between the VEZT expression levels and its clinicopathological characteristics,prognosis were also investigated.VEZT cDNA was extracted and amplificated from 293 cells by polymerase chain reaction (PCR).Using DNA recombinant technique,the target gene was connected to the pEGFP-N1 vector to construct recombinant plasmid vector after the target gene and pEGFP-N1 were purified.The recombinant plasmid was identificated by 1% enzyme electrophoresis and DNA sequencing.Results The VEZT-positive expression in gastric carcinoma was 30.3% and 60.5% in normal gastric tissues.VEZT expression in high differentiated cancer tissues was higher than that in lower differentiated tissues(x2 =5.002,P < 0.05),expression of VEZT in early gastric cancer was significantly higher than that in patients with advanced gastric cancer (x2 =5.551,P < 0.05),expression of VEZT in patients without lymph node metastasis was significantly higher than that of lymph node metastasis (x2 =5.878,P < 0.05).Patients with positive VEZT expression have better prognosis than the negative patients(x2 =6.908,P < 0.01).The target fragment,consistent with the theoretical value,was verified by gel electrophoresis.DNA sequencing confirmed that the gene sequence had no mutation and the eukaryotic expression plasmid vector pEGFP-N1-hVEZT was successfully constructed.Conclusions VEZT protein was correlated with TNM staging,tumor stage,invasion depth and lymph node metastasis,positive VEZT expression predicting better five year survival.The VEZT over-expression plasmid vector was successfully constructed.
5.Expression of p27 in Rectal Carcinoma and Modulatory Relation Between Transforming Growth Factor-?1 and p27
Changqing JING ; Leping LI ; Zhao LI ; Taihuang WU ; Qingling MU ;
Chinese Journal of Bases and Clinics in General Surgery 2004;0(01):-
Objective To detect p27 expression in rectal carcinoma and serum transforming growth factor ? 1 (TGF ? 1) level in these patients, and to elucidate the modulatory effect of serum TGF ?1 on p27 expression in rectal carcinoma. Methods Expression of p27 was measured in 37 cases of rectal carcinoma, 22 of rectal adenoma and 19 of normal control specimens by immunohistochemical staining using antibodies to p27. Serum level of TGF ?1 was measured in these patients by enzyme linked immunosorbent assay (ELISA) method. Results p27 protein was expressed in normal rectal tissue, rectal adenoma and rectal carcinoma, and the positive rate was 89.47%, 90.91% and 64.87%, respectively. The positive rate of p27 in rectal carcinoma was significantly lower than that of normal rectal tissue and rectal adenoma ( P =0.025). p27 was mainly located in nucleolus of normal rectal tissue and rectal adenoma, and the positive rate of p27 in cytoplasm of rectal carcinoma was higher than that of normal and rectal adenoma. The positives rates of serum TGF ?1 in normal group, rectal adenoma group and rectal carcinoma group were 21.05%, 27.27% and 51.35%( P =0.045),respectively. The expression of p27 related to histological differentiation, lymph node metastasis and infiltration depth. Serum level of TGF ?1 related to lymph node metastasis, infiltrated depth and CEA level. The positive rate of p27 in TGF ?1 negative group and positive group was 88.89% and 42.11%(Mantel Haenszel ? 2=6.755, P =0.009), respectively. Conclusion TGF ?1 may be useful in assessment of malignance and prognosis of rectal carcinoma. TGF ?1 can down regulate p27 expression in rectal carcinoma.
6.Expression and significance of EZH_2 and PTEN in pancreatic cancer and non-cancerous pancreatic tissues in rats
Qinglong LI ; Leping YANG ; Xiongying MIAO ; Zhulin YANG
Chinese Journal of General Surgery 1993;0(03):-
Objective To study the expressive levels of EZH2 and PTEN in pancreatic cancer and non-cancerous tissue,and the effects on carcinogenesis of rat pancreas.Methods dimethylbenzathracene(DMBA) was directly implanted into the parenchyma of rat pancreas(group A,group B).The rats of group B were treated with 1 mL trichostatin A(TSA) solution(1?g/mL) intravenously per weer 1 week after the model were set up.The rats of group A,B were killed within 3-5 months and the rats in the control(C group) were executed at 5 months to observe the develope of pancreatic cancer by macrograph and microscopy,The EnVisionTM immunohistochemistry was used to assay the expression of EZH2 and PTEN in above pancreatic specimens.Results(1) The incidence of pancreatic cancer within 3-5 months in group A was 48.7%(18/37),including 17 cases of ductal adenocarcinoma and 1 case of fibrosarcoma.The incidence of pancreatic cancer in group B was 33.3%(12/36),including 11 cases of ductal adenocarcinoma and 1 cases of fibrosarcoma.The mean maximal diameter of mass was significantly higher in group A than that in group B(P 0.05).The non-cancerous pancreatic tissues with positive expression of EZH2 and/or negative expression of PTEN showed mild to severe atypical hyperplasia of ductal epithelium.An inconsistency was found between the expression of EZH2 and PTEN in ductal adenocarcinoma(P =0.045).Pancreas of group C showed negative expression of EZH2 and positive expression of PTEN.Two cases of fibrosarcoma showed negative expression of EZH2 and PTEN.Conclusions By use of higher dose of DMBA directly implanted into the parenchyma of pancreas,high incidence of pancreatic cancer can be obtained.TSA could have an inhibitive effect on carcinogenesis and growth of pancratic cancer.The activation of EZH2 gene and inactivation of PTEN gene might have Key effects on pancreas carcinogenesis induced by DMBA in rat.
7.Comparative Study of Mammography with Ultrasonography for the Diagnosis of Breast disease with Microcalcification
Haiying WANG ; Huawei ZHANG ; Leping LI ; Hua XIN ; Qibo ZHANG
Chinese Journal of Current Advances in General Surgery 2004;0(05):-
Objective:To explore the diagnostic value of mammography and ultrasonography(US)in diagnosis of breast disease with microcalcifications.Methods:Seventy-eight cases with breast microcalcifications were detected by mammography.The detection rate of breast microcalcification at US was analyzed.And the sensitivity,specificity and the accuracy of US in diagnosis breast disease with microcalcifications was compared with these of mammography.Results:The detection rate of breast microcalcification was 66.7%,And US depicted more malignant(87.5%)than benign microcalcification(33.3%).The sensitivity specificity and accuracy of US was 68.7%、83.3%、 74.3%respectively,and the sensitivity specificity and accuracy of mammography was 72.9%、 73.3%、75.6%respectively,there were no significant difference between the two methods.The sensitivity,specificity and accuracy of the combined the two methods was 89.6%,90.0%,92.3% respectively.Conclusion:US can effectively detect and identify the breast microcalicications.The combination can improve the assessment of breast disease with microcalicications,and has a significant clinical practical value in diagnosis of early breast cancer.
8.Coexistence of Gitelman's syndrome and thyroid disease: SLC12A3 gene analysis in two patients
Hui DONG ; Yanhua LANG ; Zeping SHAO ; Lin LI ; Leping SHAO
Chinese Journal of Endocrinology and Metabolism 2010;26(5):395-398
Two patients with coexistence of thyroid disease and suspected Gitelman's syndrome underwent SLC12A3 gene analysis. The results confirmed that both patients were compound heterozygotes of SLC12A3 gene mutation. Three novel variants of SLC12A3 were found in this study. This report suggests that Gitelman's syndrome may coexist with other disorders associated with hypokalemia, such as Graves' disease.
9.Protective effect of astaxanthin on endothelial progenitor cell apoptosis induced by oxidative stress
Pei LI ; Qiongyu ZHANG ; Leping ZENG ; Zhigang GONG
Chongqing Medicine 2014;(26):3464-3467
Objective To investigate the effect of astaxanthin on the peripheral blood endothelial progenitor cells (EPCs) apop-tosis induced by oxidative stress in vitro and to explore its underlying mechanism .Methods Human peripheral blood EPCs were in vitro cultured and divided into the control group ,model group with 100 μmol/L tert-butyl hydroperoxide(tBHP) and the astaxan-thin plus tBHP group(with 0 .10 ,1 .00 ,10 .00 nmol/L astaxanthin pretreatment for 24 h ,then adding the final concentration of 100μmol/L tBHP for 6 h continuous culture) .The cell viability was measured by the MTT method .The level of reactive oxygen spe-cies (ROS) was determined by the DCFH-DA method .The changes of mitochondrial membrane potential(MMP) and the apoptosis ratio were detected by the JC-1 method and the DAPI method ,respectively .Results Compared with control group ,100μmol/L tB-HP could obviously caused the apoptosis of EPCs(P<0 .05) ,while astaxanthin could decrease tBHP induced apoptosis ,which man-ifested by the decrease of the apoptosis ratio (P<0 .05) and MMP increase .Conclusion Astaxanthin has the protective effect on the apoptosis of EPCs ,its mechanism may be related with the protection of the mitochondrial function .
10.Signal noise filtering of patch clamp based on Wavelet transform
Changhai LEI ; Yongji YANG ; Dongfang LI ; Leping ZHANG
Chinese Medical Equipment Journal 2003;0(11):-
Wavelet transform(WT) is a powerful technique in signal separation and can improve the signal-to-noise ratio by separating the white noise and useful signal.The method in the paper introduces the concepts of Wavelet transform.According to the analysis of Wavelet coefficients between the noise and signal at different levels,the appropriate filter is realized.The patch clamp experiment results demonstrate the feasibility of the method.