1.PRELIMINARY STUDY TO THE PROGRAMMED CELL DEATH OF HL-60 CELL AND BGS-180 CELL INDUCED BY VP-16
Medical Journal of Chinese People's Liberation Army 2001;26(2):117-118
Programmed cell death(PCD)of human leukemic HL-60 cell and human poorly differentiated gastric adenocarcinoma cell BGS-180 induced by efoposid(VP-16) was preliminarily observed comparatively in the same experimental condition through rate of cell death, DNA agarose gels electrophoresis and terminal deoxynucleotidyl transferase(TdT) mediated biotin-11-dUTP nickend labeling(TUNEL).It was found that apoptosis is the main pattern of HL-60 cell death induced by low does VP-16 in a short time, and it can be suppressed when protein kinase C(PKC) is activated. The main pattern of BGS-180 cell death induced by VP-16 is necrosis, and PCK activation does not affect its necrosis rate. Extracellular Ca2+ reduction do not affect BGS-180 and HL-60 cell death rate. The mechanism of VP-16 action on BGS-180 and HL-60 cell is different, apoptosis is not the main pattern of cell BGS-180 death induced by VP-16.
2.PRELIMINARY STUDY TO THE PROGRAMMED CELL DEATH OF HL-60 CELL AND BGS 180 CELL INDUCED BY VP-16
Medical Journal of Chinese People's Liberation Army 1983;0(02):-
Programmed cell death(PCD)of human leukemic HL 60 cell and human poorly differentiated gastric adenocarcinoma cell BGS 180 induced by efoposid(VP 16) was preliminarily observed comparatively in the same experimental condition through rate of cell death, DNA agarose gels electrophoresis and terminal deoxynucleotidyl transferase(TdT) mediated biotin 11 dUTP nickend labeling(TUNEL).It was found that apoptosis is the main pattern of HL 60 cell death induced by low does VP 16 in a short time, and it can be suppressed when protein kinase C(PKC) is activated. The main pattern of BGS 180 cell death induced by VP 16 is necrosis, and PCK activation does not affect its necrosis rate. Extracellular Ca 2+ reduction do not affect BGS 180 and HL 60 cell death rate. The mechanism of VP 16 action on BGS 180 and HL 60 cell is different, apoptosis is not the main pattern of cell BGS 180 death induced by VP 16.
3.ANALYSIS OF RELEVANT FACTORS OF MALIGNANT DEGENERATION OF COLORECTAL POLYPS
Zhongsheng LU ; Yunsheng YANG ; Xueme ZHUANG
Medical Journal of Chinese People's Liberation Army 2001;0(08):-
Objective To retrospectively analyse the factors related to the risk of malignant change in relation with various colorectal polyps. Methods Data on a total of 820 patients (900 polyps), who had undergone polypectomy between 1997 and 2001 were collected. The relation between colorectal cancer and polyps characteristics was assessed by Logistic Regression and Chi square. Results The risk of malignant change was significantly related with the size of polyps, histology of polyps, grade of dysplasia, pathological changes of mucosa, sessile or pedunculated polyps. Conclusion High grade dysplasia, large size, adenoma, absence of a stalk, and ulcerous mucosa were confirmed as the major predictors. The likelihood of malignant degeneration of colorectal polyps seemed to be correlated with the number of the above mentioned risk factors present
4.The diagnostic value of confocal laser endomicroscopy in colorectal polyps
Mingyang LI ; Min ZHU ; Zhiqiang WANG ; Jin HUANG ; Zhongsheng LU
Chinese Journal of Digestion 2010;30(11):803-807
Objective To analyze endoscopic histological characters and establish diagnostic standards of colorectal polyps with confocal laser endomicroscopy (CLE) examination and to explore the diagnostic value of the CLE in adenomatous polyps and non-adenomatous polyps. Methods From June to December in 2009, 90 patients were recruited in this study, which included 40 pathologically confirmed colorectal polyps patients (total 48 colon and rectal polyps) and 50 patients for prospective study (total 106 colon rectal polyps). At same time 10 spots of normal mucosa was taken for comparison. Firstly the CLE images of 48 pathologically confirmed colorectal polyps (22 adenomatous polyps and 26 non-adenomatous polyps) were analyzed, and the diagnosis criteria for distinguishing adenomatous polyps and non-adenomatous polyps was established with CLE. Then according to the criteria, 106 colorectal polyps underwent prospective diagnosis with CLE. Finally, the CLE diagnosis result was compared with pathologically result to evaluate the diagnosis efficiency of CLE. Results In the 48 colorectal polyps of 40 pathologically confirmed colorectal polyps patients, there were 22 adenomatous polyps and 26 non-adenomatous polyps. The sensitivity, specificity and accuracy of CLE in adenomatous polyps diagnosis were 94. 0%, 92. 9%, and 93. 4%, respectively. The positive predictive value was 92.2% and the negative predictive value was 94.5%. The sensitivity, specificity and accuracy of CLE in non-adenomatous polyps diagnosis were 92. 9%, 94.0%, and 93. 4%respectively, the positive predictive value was 94.5% and the negative predictive value was 92.2%.The coherence of CLE and histopathology in adenomatous polyps diagnosis was pretty good (Kappa=0. 893). Conclusion The accuracy of CLE in adenomatous polyps and non-adenomatous polyps differential diagnosis was high, and the coherence with histopathology diagnosis was good, which provided experience for further detection of early rectal precancerous.
5.Construction of Recombinant Plasmid of Human RIP3 Gene and Its Effects on the Death of Breast Cancer MCF7 Cells
Can LU ; Huijun XU ; Yongsheng JIA ; Zhongsheng TONG
Tianjin Medical Journal 2014;(2):109-112
Objective To construct the recombinant RIP3 over-expressed plasmids and transfect them in breast cancer MCF7 cells, and identify the expression and localization of fusion protein, as well as its effect on the death way of MCF7 cells. Methods The expression levels of RIP3 mRNA in four breast cancer cell lines and normal mammary epithelial cells were detected by reverse transcription polymerase chain reaction (RT-PCR). The RIP 3 coding sequence was amplified by polymerase chain reaction and subcloned into mCherry vector to construct recombinant plasmids. The plasmids were transfected into MCF7 cells by lentivirus after DNA sequencing, then screened by basticidin (4 mg/L) for 1 week. The efficiencies of RIP3 expression were validated by Western blotting assay. The death way of mCherry-RIP3-MCF7 cells was observed under the treatment of TNF-αand Z-VAD-FMK. Results The lowest expression of RIP3 mRNA was found in MCF7 cells. The sequencing results validated the well recombinant plasmids. The expression of mCherry-RIP3 fusion pro-tein with a molecular weight of 85 ku was detected by Western blot assay. The mCherry-RIP3 expression enhanced the sensi-tivity of MCF7 cells to TNF-αand Z-VAD-FMK induced cell death. Conclusion The recombinant RIP3 over-expressed plasmids were successfully constructed, and the stable MCF7 cells with ectopic RIP3 transfection were obtained. The mCher-ry-RIP3 fusion protein was expressed in the cytoplasm and was conformed to mediate TNF-αinduced necroptosis.
6.Ulinastatin Combined with Xingnaojing Injection in Treatment of Acute Cerebral Hemorrhage and Serum hs-CRP, D-D and NSE Levels
Zhongsheng LU ; Lichao ZHANG ; Peng YANG ; Qiang ZHANG ; Xiaojie CONG
Progress in Modern Biomedicine 2017;17(24):4719-4722
Objective:To investigate ulinastatin (UTI) combined with Xingnaojing injection in the treatment of acute cerebral hemorrhage (ACH) and its effect on the serum high sensitivity C reactive protein (hs-CRP),D-dimer (D-D) and neuron specific enolase(NSE) levels.Methods:110 cases of ACH patients admitted in our hospital from January 2015 to December 2016 were selected and divided into two groups according to the random number table method.The control group was given UTI treatment,while the observation group was given UTI combined with Xingnaojing treatment.Then the brain edema absorption effect,NIHSS score,serum hs-CRP,D-D and NSE levels before and after the treatment of the two groups were recorded and compared;the safety ofmedicidstion of the two groups was evaluated.Results:At the 14th day after treatment,the total effective rate of cerebral hematoma absorption in the observation group was 89.1%,which was significantly higher than 67.3% of the control group (P<0.01).At the 14th day after treatment,the NIHSS scores of both groups were significantly lower than those before the treatment (P<0.01);compared with that of the control group of the same time period,at the 14th day after treatment,the improvement effect of NIHSS score in the observation group was more significant (P<0.01).Compared with those before the treatment,the serum hs-CRP,D-D and NSE levels of both groups at the 14th day after treatment were significantly decreased (P<0.01);at the 14th day after treatment,the serum indicators of the observation group improved more significantly than those of the control group (P<0.01).The incidence rate of adverse reaction in the observation group was 3.6% compared with 5.5% of the control group (P>0.05).Conclusion:Ulinastatin combined with Xingnaojing Injection could rapidly relieve or eliminate hematoma in the treatment of acute cerebral hemorrhage,control the inflammatory response,improve the blood coagulation system and fibrinolytic system,protect the nerve cells and reduce the neurological damagee.
7.Effect of targeted inhibition of occludin by microRNA-429 on intestinal epithelial barrier function in diabetic mouse
Xiji LU ; Tao YU ; Zhongsheng XIA ; Tidong SHAN ; Hui OUYANG ; Jihao XU ; Qikui CHEN
Chinese Journal of Pathophysiology 2016;32(4):696-700
AIM:To investigate the effect of microRNA-429 (miR-429) on the expression of occludin (Ocln) in intestinal epithelial cells ( IECs) and intestinal epithelial barrier function in diabetic mice.METHODS:Diabetes mel-litus mouse model was induced by intraperitoneal injection of streptozocin.The expression of miR-429 in IECs of diabetic mice was inhibited by antagomiRNA-429 injected via tail vein.The expression of miR-429 and mRNA expression of Ocln were detected by real-time PCR.The protein expression of Ocln was determined by Western blotting and immunohistochem-istry.The urinary lactulose/mannitol ratio was measured by gas chromatography.The plasma LPS concentrations in the mice were measured by chromogenic end-point TAL kit.RESULTS:The results of real-time PCR confirmed that the ex-pression of miR-429 in IECs of diabetic mice was remarkably inhibited by tail-vein administration of antagomiRNA-429, and resumed to similar level of normal mice on the 6th day after the administration.After suppressing the level of miR-429, the expression of Ocln in IECs of diabetic mice increased significantly (P<0.05), while the urinary lactulose/mannitol ra-tio and the plasma LPS concentration decreased obviously ( P<0.05 ) .CONCLUSION:AntagomiRNA-429 effectively suppresses miR-429 expression in IECs of diabetic mice, and then enhances the expression of Ocln and partially resumes the intestinal epithelial barrier function.
8.Effect of Wip1 gene silencing on chemotherapy sensitivity of human colon cancer cells
Di WU ; Zhongsheng XIA ; Wa ZHONG ; Xiji LU ; Tao YU ; Qikui CHEN
Chinese Journal of Pathophysiology 2015;(5):857-863
AIM:To observe the inhibitory effect of siRNA targeting to Wip1 gene on the Wip1 gene expression in the colon cancer cells and to investigate the influence of Wip1 gene silencing on the chemotherapy sensitivity of colon cancer cells.METHODS:Wip1-811 siRNA targeting to Wip1 gene was transfected into RKO colon cancer cells with high expression of Wip1 gene.The mRNA expression of Wip1 was measured by real-time PCR.The protein level of Wip1 was detected by Western blotting.The viability of RKO colon cancer cells was measured by MTS assay.The cell apoptosis and cell cycle were analyzed by flow cytometry.RESULTS: Wip1-811 siRNA efficiently inhibited the expression of Wip1 at mRNA and protein levels.The enhanced chemotherapy sensitivity of RKO colon cancer cells was observed after inhibition of Wip1 gene expression.The viability of RKO colon cancer cells was decreased from (89.4 ±6.6)%to (74.7 ±3.9)%af-ter treated with 5-fluorouracil (P<0.05) and decreased from (77.9 ±2.4)%to (66.7 ±2.9)%after treated with oxali-platin ( P<0.05 ) .The cell apoptotic rate was increased from ( 7.7 ±0.5 )% to ( 12.3 ±3.2 )% and from ( 14.7 ± 2.1)% to (34.0 ±2.1)% when RKO colon cancer cells were treated with 5-fluorouracil and oxaliplatin, respectively (P<0.05).CONCLUSION:Wip1 gene silencing enhances chemotherapy sensitivity of colon cancer cells.
9.Clinical features and prognostic analysis of rectal neuroendocrine neoplasm in different pathological grades
Man MENG ; Enqiang LINGHU ; Po ZHAO ; Zhongsheng LU ; Zhiqiang WANG ; Yonghua WANG ; Qiyang HUANG
Chinese Journal of Digestive Surgery 2014;13(10):789-792
Objective To investigate the clinical features and prognosis of rectal neuroendocrine neoplasms (NENs) in different pathological grades.Methods The clinical data of 183 patients with rectal NENs who were admitted to the PLA General Hospital from January 2001 to April 2012 were retrospectively analyzed.All the clinical and pathological data of the patients who received endoscopy and (or) surgical resection were retrieved from the work station and the database of the endoscopic center.Based on the 2010 WHO pathology classification of digestive tumors,the pathological data were ranked according to the mitotic count.The prognosis of the patients was learned by re-examination or phone call.The follow-up ended till July 2014 or at the death of patients.Data were analyzed using the chi-square test.Results A total of 183 patients were enrolled in this study including 120 males and 63 females.The median age of the patients was 48 years (range,14-83 years).Seventy-four patients had the symptom of hemafecia,9 patients had abdominal pain and change in bowel habit,and 70 patients were diagnosed by body examination.Other symptoms included increased level of tumor markers and abdominal distension.Carcinoid syndrome was not detected in all the patients.Fourteen patients were complicated with polyp of intestine,5 with tubular adenoma,3 with colorectal adenocarcinoma and 1 with small cell lung cancer.The diameters of the tumor under 1 cm were detected in 162 patients,the diameters of the tumors ranged between 1 cm and 2 cm in 14 patients,and the diameters of the tumors above 2 cm in 7 patients.The mean distance between the tumor and the anus was (5-± 3)cm.Of the 183 patients,130 received endoscopic treatment,43 received surgical treatment and 10 received clamping because the tumor was misdiagnosed as polyps.There were 158 patients in grade1 (154 in stage Ⅰ,1 in stage Ⅱ,1 in stage Ⅲ and 2 in stage Ⅳ),21 in grade2 (13 in stage Ⅰ,3in stage Ⅱ,3 in stage Ⅲ and 2 in stage Ⅳ),4 in grade 3 (1 in stage Ⅰ,1 in stage Ⅲ and 2 in stage Ⅳ).Six patients had liver metastasis and 9 had lymph node metastasis.Fourteen patients died (4 in grade 1,6 in grade 2 and 4 in grade 3).The 5-year survival rate of patients was 92.35% (169/183).There were significant differences in the gender,tumor diameter,tumor staging,lymph node metastasis distal metastasis and 5-year survival rate among patients with rectal NENs in different pathological grades (x2=60.949,71.587,32.135,55.486,56.512,P < 0.05).Conclusions Rectal NENs lacks the specific clinical manifestation and is more likely to happen in males,and it often locates at the middle-lower rectum.Most of the rectal NENs belongs to stage Ⅰ and grade 1 and is less than 1 cm in size.The prognosis of patients with rectal NENs in different pathological grades is different.The 2010 WHO pathology classification of digestive tumors is useful to asses the prognosis of rectal NENs.Different grades of rectal NENs could be taken into account when designing the treatment plan.
10.The value of esophageal intrapapillary capillary loop visualized by magnifying narrow-band imaging endoscopy in diagnosing esophageal mucosal pathology
Shufang WANG ; Yunsheng YANG ; Jing YUAN ; Xiuli ZHANG ; Zhongsheng LU ; Gang SUN ; Enqiang LINGHU ; Jiangyun MENG
Chinese Journal of Internal Medicine 2012;51(4):284-288
Objective To investigate the diagnostic potential of magnifying narrow-band imaging endoscopy (NBI-ME) for different intrapapillary capillary loop (IPCL) for the diagnosis of esophageal lesion.Methods Patients with abnormal esophageal mucosa found by white light gastroscopy in digestive endoscopy center,Chinese PLA General Hospital during the period of November 2009 to November 2010 were enrolled in this study.IPCL was observed and divided into different types by NBI-ME.Histopathology of biopsy or endoscopic submucosal dissection (ESD) specimens was evaluated and used as the gold standard to evaluate the diagnostic value of NBI-ME for IPCL.Results A total of 146 lesions from 145 subjects with esophageal mucosa abnormal were collected. Among them, 88 were pathology-proven inflammation,5 were pathology-proven esophageal cancers,20 were pathology-proven low intraepithelial neoplasia (LIN) and 33 were pathology-proven high intraepithelial neoplasia (HIN) detected with NBI-ME.By a per-lesion analysis,the accuracy of inflammation and cancer were 100% (88/88) and 7/7.For the sensitivity,specificity,accuracy,positive predictive value,negative predictive value,positive likelihood ratio,negative likelihood ratio of LIN and HIN were 7/10,69.8% ( 30/43 ),69.8% ( 37/53 ),35.0% (7/20),90.9% (30/33),12.5% (70/559),2.3% (30/1290) and 87.1% (27/31),72.7% ( 16/22),81.1% ( 43/53 ),81.8% ( 27/33 ),80.0% ( 16/20 ),634.1% ( 837/132 ) and 35.2% ( 124/352 ),respectively.Conclusions NBI-ME can classify the different esophageal IPCL.Higher diagnostic accuracy of IPCL indicates the feasibility of NBI-ME for the efficacious diagnosis of esophageal inflammation and cancer.There is the higher diagnostic accuracy of HIN than LIN.