1.HIGH-LEVEL EXPRESSION OF RECOMBINANT TRICHOSANTHIN IN ESCHERICHIA COLI AND EVALUATION OF ITS IMMUNOGENICITY
Ping WANG ; Shiping DING ; Jicheng LI
Acta Anatomica Sinica 1954;0(02):-
Objective The TCS gene without N-and C-terminal sequences was cloned into pET-29b in order to get the recombinant trichosanthin(rTCS) at the relatively high level in E.coli,and the rTCS immunogenicity was compared with natural TCS(nTCS). Methods TCS gene without the N-and C-terminal coding sequences was amplified by RT-PCR,and then inserted into pET-29b.The soluble rTCS was induced with IPTG and purified through metal chelate affinity chromatography,the purity and content were showed by the thin-layer scan analysis and bradford assay.After the rabbits were immunized with purified rTCS and nTCS,respectively,the titer and specificity of polyclonal antibody were detected by indirect ELISA assay and Western blotting. Results 1.The vector pET-29b-TCS was constructed successfully after it was confirmed with the same restriction enzymes.2.The rTCS attained the maximum in the presence of 1 mmol/L IPTG for 4?h at 30℃.3.The soluble rTCS was expressed at relatively high level(36% of total protein in E.coli) and the yield of rTCS with a purity of 95% was 1.2?g in 1 L bacterium.4.The polyclonal antisera titer of nTCS was significantly higher than that of rTCS(1∶40?960 and 1∶5?120 respectively).5.The specific reaction between rTCS and anti-nTCS antibody was detected by Western blotting.Conclusion The high level expressing vector pET-29b-TCS,carrying TCS gene withoug the N-and C-terminal coding sequences,was constructed successfully.The immunogenicity of rTCS was remarkably lower than that of nTCS,which indicates that glycosylation of nTCS has a relationship with the antigenicity.
2.Effect of Ershen Zeshu Mixture on the peritoneal lymphatic stomata and the ascites drainage
Shiping DING ; Jicheng LI ; Liangen MAO ; Jian XU ; Jiandon LIU
Chinese Journal of Pathophysiology 2000;0(11):-
AIM: To further study the effect of Ershen Zeshu Mixture, a Chinese medicine, on the peritoneal lymphatic stomata and the ascites drainage in experimental mice model of CCl 4-induced fibrosis.By the application of NO supplier (sodium nitroprusside,SNP) and NO suppressor (N G-mononethyl-L-arginine,L-NMMA),the effect of NO on the peritoneal lymphatic stomata and the ascites drainage was studied to clarify the relationship between Ershen Zeshu Mixture and NO in the regulation of the peritoneal lymphatic stomata.METHODS: The mouse model of fibrosis was established with the application of intragastric installations of carbon tetrachloride one time every three days;Scanning electron microscope and computer image processing were used to detect the girth,area and distribution density of the peritoneal lymphatic stomata;the concentration of urinary ions was measured in the experiment.RESULTS: ①Compared to NS group and model group,Ershen Zeshu Mixture could significantly increase the girth and the area of the peritoneal lymphatic stomata,promote its distribution density ( P
3.Gene polymorphism in chromosome 7q35 and susceptibility to diabetic nephropathy
Yanhu DONG ; Shiping QU ; Wenshan LV ; Ming DING ; Chao DONG ; Hongwei JIANG ; Jie CHENG
Chinese Journal of Endocrinology and Metabolism 1985;0(01):-
Objective To investigate the relationship between the polymorphisms of both aldose reductase (AR) and endothelial nitric oxide synthase (eNOS) genes in chromosome 7q35 and the susceptibility to diabetic nephropathy (DN) in type 2 diabetes of Han nationality in North China. Methods The C(-106)T in the promoter region of AR gene as well as the G894T in exon 7 and the 27 bp repeat polymorphism in intron 4 of the eNOS gene were investigated in 85 healthy controls and 134 type 2 diabetics with or without DN. The C(-106)T as well as the G894T genotype were determined by PCR RFLP method and sequencing the PCR products. The 27 bp repeat polymorphism alleles were determined by PCR combined with agarose gel electrophoresis and sequencing the PCR products. Results The frequencies of the C allele and C/C genotype in AR gene as well as T allele and T/G genotype in exon 7 and the 4a allele and 4a/4b genotype in intron 4 of eNOS gene were significantly higher in DN+ group than those in DN-group (all P
4.Epidemiological characteristic and current status of surgical treatment for esophageal cancer by analysis of national registry database
Yousheng MAO ; Shugeng GAO ; Qun WANG ; Xiaotian SHI ; Yin LI ; Wenjun GAO ; Fushun GUAN ; Xiaofei LI ; Yongtao HAN ; Yongyu LIU ; Junfeng LIU ; Kang ZHANG ; Shuoyan LIU ; Xiangning FU ; Wentao FANG ; Longqi CHEN ; Qingchen WU ; Gaoming XIAO ; Keneng CHEN ; Guanggen JIAO ; Shijiang ZHANG ; Weimin MAO ; Tiehua RONG ; Jianhua FU ; Lijie TAN ; Chun CHEN ; Shidong XU ; Shiping GUO ; Zhentao YU ; Jian HU ; Zhendong HU ; Yikun YANG ; Ningning DING ; Ding YANG ; Jie HE
Chinese Journal of Oncology 2020;42(3):228-233
Objective:To investigate the epidemiological characteristics and current status of surgical management for esophageal cancer in China.Methods:A national database was setup through a network platform. The clinical data of esophageal cancer treated by surgery was collected from 70 major hospitals in China between January 2009 and December 2014.Results:Complete data of 8 181 cases of esophageal cancer patients who underwent surgery were recorded in the database and recruited in the analysis. Among them, 6 052 cases were male and 2 129 were female, the average age was 60.5 years.The epidemiological investigation results showed that 148 cases (1.8%) had history of psychological trauma, 7 527 cases (92.0%) were lower social economic status, 5 072 cases (62.0%) were short of fresh vegetables and fruits, 6 544 cases (80.0%) ate rough food frequently, 3 722 cases (45.5%) drank untreated water directly from lake or river or shallow well, 3 436 cases (42.0%) had a unhealthy eating habit, including habits of eating food fast (507 cases, 6.2%), eating hot food or drinking hot tea/soup (998 cases, 12.2%), eating fried food (1 939 cases, 23.7%), 4 410 cases (53.9%) had the habits of smoking cigarettes and 2 822 cases (34.5%) drank white wine frequently.The pathological results showed that 7 813 cases (95.5%) were squamous cell carcinoma, 267 cases were adenocarcinoma (3.3%), 25 cases were adenosquamous cell carcinoma (0.3%) and 50 cases were small cell carcinoma (0.6%). A total of 1 800 cases (22.0%) received preoperative neoadjuvant therapy due to locally advanced disease or difficulty of resection. The esophagectomies were performed through left thoracotomy approach in 5 870 cases (71.8%), through right chest approach in 2 215 cases (27.1%), and the remain 96 cases (1.2%) received surgery though other approaches.A total of 8 001 cases (97.8%) underwent radical resection, the other 180 cases (2.2%) received palliative resection. The 30-day postoperative mortality rate was 0.5%, the overall ≥ grade Ⅱ postoperative complication rate was 11.6% (951 cases). The 1-yr, 3-yr, and 5-yr overall actual survival rates were 82.6%, 61.6%, and 52.9%, respectively.Conclusions:The data analysis of the national database for esophageal cancer shows that bad eating habits or eating rough food without enough nutrients, lower social and economic status, drinking white wine and smoking cigarettes frequently may be correlated with tumorigenesis of esophageal cancer. However, strong evidences produced by prospective observation studies are needed. Overall, the long-term survival of esophageal cancer patients has been improved gradually due to the application of advanced surgical techniques and reasonable multimodality treatment.
5.Epidemiological characteristic and current status of surgical treatment for esophageal cancer by analysis of national registry database
Yousheng MAO ; Shugeng GAO ; Qun WANG ; Xiaotian SHI ; Yin LI ; Wenjun GAO ; Fushun GUAN ; Xiaofei LI ; Yongtao HAN ; Yongyu LIU ; Junfeng LIU ; Kang ZHANG ; Shuoyan LIU ; Xiangning FU ; Wentao FANG ; Longqi CHEN ; Qingchen WU ; Gaoming XIAO ; Keneng CHEN ; Guanggen JIAO ; Shijiang ZHANG ; Weimin MAO ; Tiehua RONG ; Jianhua FU ; Lijie TAN ; Chun CHEN ; Shidong XU ; Shiping GUO ; Zhentao YU ; Jian HU ; Zhendong HU ; Yikun YANG ; Ningning DING ; Ding YANG ; Jie HE
Chinese Journal of Oncology 2020;42(3):228-233
Objective:To investigate the epidemiological characteristics and current status of surgical management for esophageal cancer in China.Methods:A national database was setup through a network platform. The clinical data of esophageal cancer treated by surgery was collected from 70 major hospitals in China between January 2009 and December 2014.Results:Complete data of 8 181 cases of esophageal cancer patients who underwent surgery were recorded in the database and recruited in the analysis. Among them, 6 052 cases were male and 2 129 were female, the average age was 60.5 years.The epidemiological investigation results showed that 148 cases (1.8%) had history of psychological trauma, 7 527 cases (92.0%) were lower social economic status, 5 072 cases (62.0%) were short of fresh vegetables and fruits, 6 544 cases (80.0%) ate rough food frequently, 3 722 cases (45.5%) drank untreated water directly from lake or river or shallow well, 3 436 cases (42.0%) had a unhealthy eating habit, including habits of eating food fast (507 cases, 6.2%), eating hot food or drinking hot tea/soup (998 cases, 12.2%), eating fried food (1 939 cases, 23.7%), 4 410 cases (53.9%) had the habits of smoking cigarettes and 2 822 cases (34.5%) drank white wine frequently.The pathological results showed that 7 813 cases (95.5%) were squamous cell carcinoma, 267 cases were adenocarcinoma (3.3%), 25 cases were adenosquamous cell carcinoma (0.3%) and 50 cases were small cell carcinoma (0.6%). A total of 1 800 cases (22.0%) received preoperative neoadjuvant therapy due to locally advanced disease or difficulty of resection. The esophagectomies were performed through left thoracotomy approach in 5 870 cases (71.8%), through right chest approach in 2 215 cases (27.1%), and the remain 96 cases (1.2%) received surgery though other approaches.A total of 8 001 cases (97.8%) underwent radical resection, the other 180 cases (2.2%) received palliative resection. The 30-day postoperative mortality rate was 0.5%, the overall ≥ grade Ⅱ postoperative complication rate was 11.6% (951 cases). The 1-yr, 3-yr, and 5-yr overall actual survival rates were 82.6%, 61.6%, and 52.9%, respectively.Conclusions:The data analysis of the national database for esophageal cancer shows that bad eating habits or eating rough food without enough nutrients, lower social and economic status, drinking white wine and smoking cigarettes frequently may be correlated with tumorigenesis of esophageal cancer. However, strong evidences produced by prospective observation studies are needed. Overall, the long-term survival of esophageal cancer patients has been improved gradually due to the application of advanced surgical techniques and reasonable multimodality treatment.