4.Survioin promoter activity in tumor cell lines
rang, XU ; sheng-fang, GE ; jian, LU ; guan-xiang, QIAN
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(08):-
Objective To study the activity of the survivin gene promoter in several tumor cell lines and evaluate the possible application of this promoter in tumor gene therapy. Methods ①The expressions of survivin gene in A549,MDA-MB231 and HepG2 cell lines were detected by RT-PCR and Western blotting.②Tumor cells(A549,MDA-MB231,HepG2) were transiently transfected by reporter plasmids containing different length of survivin promoter using lipofectamine.And 48 h later,the level of reporter gene expression was analyzed.Results There were different levels of survivin expression in A549,MDA-MB231 and HepG2 cell lines.Transient transfection assay approved that pLuc-surP-987,pLuc-surP-596,pLuc-surP-269 and pLuc-surP-158 showed high activity and 269 bp survivin promoter demonstrated the highest activity. Conclusion In transcriptional level,survivin promoter can activate the reporter gene in several tumor cell lines.It is a potential candidate promoter in tumor gene therapy.
5.Clinical Study on Infantile Congenital and Non-Congenital Cytomegalovirus Infection
rong, MI ; fang-sheng, XU ; cong-le, ZHOU
Journal of Applied Clinical Pediatrics 2006;0(22):-
Objective To learn the damage on infants and effect of treatment of congenital cytomegalovirus(CMV) infection and non-congenital cytomegalovirus infection which included perinatal infection and postnatal infection.Methods According to the diagnosis standard of cytomegalovirus infection in China,46 infants diagnosed CMV infection were retrospectively reviewed.In this research,CMV infection was diagnosed if serum CMV-IgM or CMV-pp65 in polymorphous leukocytes was positive.According to beginning time of CMV infection symptom,46 cases were divided into 2 groups: congenital infection and non-congenital infection,clinical features of 2 groups were compared.Ganciclovir was given to the infants with infantile hepatic syndrome of 2 groups,and compared liver transaminases and bilirulin with itself after treatment,side-effect was observed.Infants of 2 groups were followed up,and prognosis was compared.Results In 46 infants,21 were enrolled congenital infection group,25 were non-congenital infection group.CMV-IgM was positive in 38 cases of 46 patients,CMV-pp65 was positive in 19 cases of 43 patients,both CMVIgM and CMV-pp65 were positive in 11 cases.More congenital malformation were found in congenital group than non-congenital group,there was significant difference between 2 groups((P
6.The Study of Orientations of Bilirubin and Biliverdin by Surface-enhanced Raman Spectroscopy
Hu Jun ; Fang Qing ; Sheng Rongsheng ; Xu Zhisan ; ZENG Yune
Chinese Journal of Analytical Chemistry 2001;29(5):507-510
The surface-enhanced Raman spectra of bilirubin and biliverdin were obtained.By the bands analysis of the spectra,the orientation of bilirubin and biliverdin on the surface of silver colloid was discussed.In such case,the bilirubin was adsorbed on the silver colloid particle with the two planar pyrromethenone groups intercalated into the globe silver colloid particle,however,the biliverdin might lie flat on the surface of silver colloid with syn-synsyn conformation.
8.Assessment of mild chronic pancreatitis:the utility of diffusion weighted imaging before and after secretin stimulation
Yun BIAN ; Xu FANG ; Yu SHENG ; Xiao LI ; Jianping LU ; Chengwei SHAO ; Li WANG ; Zhang SHI ; Fang LIU ; Ri LIU
Chinese Journal of Radiology 2017;51(1):23-27
Objective To explore the diagnostic value of DWI after secretin stimulation for the diagnosis of mild chronic pancreatitis (CP). Methods This was a prospective study. Ninety-nine consecutive individuals including 23 healthy volunteers, 11 risk volunteers, 15 mild CP patients, 14 moderate CP patients and 36 severe CP patients underwent secretin DWI and faecal elastase 1(FE-1) testing. The subjects were grouped by Cambridge classification about endoscopic retrograde cholangiography (ERCP), CT and ultrasonography. Secretin stimulated diffusion weighted imaging(S-DWI), the ADCs, time to peak ADCs and FE-1 were performed on all subjects. The changes of pancreatic ADC values were observed before and after the injection of secretin. All ADCs and FE-1 were compared between groups with single factor analysis of variance, and the correlation between ADCs and FE-1 was determined with Pearson analysis. ROC curves were performed to identify the diagnostic efficacy of DWI related measures. Results Eight patients with severe CP were excluded because the significant atrophy of the pancreatic parenchyma prohibited the evaluation of ADC measurement. Ninety-one individuals were divided into five groups including 23 healthy volunteers, 11 risk volunteers, 15 mild CP patients, 14 moderate CP patients and 28 severe CP patients. The mean baseline and peak ADCs were higher in the healthy volunteers than in other groups, with significant differences (P<0.05). There was no ADC peak in severe CP patients. There were significant differences between the mean baseline ADCs and the peak ADCs in the other groups (P<0.05). The mild and moderate CP groups showed a delayed peak. The area under curve (AUC) of the mean baseline and peak ADCs, time to peak ADCs for differentiating mild CP was 0.818, 0.912 and 0.965, respectively. Using 4.67 min as the cutoff value, time to peak ADCs were most accurate for differentiating healthy from risk patients and those with evident pancreatitis, yielding a sensitivity of 80.0%and a specificity of 100.0%. Good correlations between baseline and peak ADCs, time to peak ADCs, and FE-1 were shown(r=0.57, 0.72 and-0.84, P<0.01). Conclusions Using the peak and time to peak ADCs may improve the detection of risk and mild CP. Secretin-enhanced DWI is a noninvasive, convenient and accurate method.
9.Construction of eukaryotic recombinant expression plasmids with glyceraldehydes-3-phosphate dehydrogenase and cysteine protease inhibitor gene of periodic Brugia malayi and its expression in HeLa cells
Xiao-jun, LIU ; Xiao-feng, GUO ; Sai-nan, ZHANG ; Shi-juan, LU ; Hao, FANG ; Bang-sheng, XU ; Zheng, FANG
Chinese Journal of Endemiology 2011;30(4):371-375
Objective To construct the eukaryotic expression plasmid containing glyceraldehydes-3-phosphate dehydrogenase (GAPDH) and cysteine protease inhibitor ( CPI ) gene from periodic Brugia malayi (Bm) and to lay foundation for studying multivalent vaccines. Methods Total RNA was extracted from periodic Bin. The BmGAPDH and BmCPI genes were amplified by RT-PCR. The PCR product was cloned and then subeloned into eukaryotic recombinant plasmid vector pcDNA3.1 (+). pcDNA3.1 (+)/BmGAPDH/BmCPI was constructed. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification, and were transformed into HeLa cell subsequently. The transient expression of BmGAPDH and BmCPI were examined by RT-PCR. The expressed protein was identified by sodium dodeeylsulphate-polyacrylamide gel electrophoresis(SDS-PAGE). Results Two specific bands of around 877 bp of BmGAPDH and 621 bp of BmCPI were amplified, consistent with the expected value. The same bands were obtained by double restriction enzyme digestion of recombinant plasmids or PCR using recombinant plasmid as template. BmGAPDH and BmCPI mRNA were highly expressed in transfeeted HeLa cell. The relative molecular mass (Mr) of the recombinant protein was about 54 × 103. Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1 (+)/BmGAPDH/BmCPI has been constructed successfully and the protein is expressed correctly in mammalian cell.
10.Kidney yang deficiency constitution and hypothalamic-pituitary-adrenal axis dysfunction in Lewis rats
Qiyang SHOU ; Lizong ZHANG ; Yueqin CAI ; Peng FANG ; Linhui NI ; Jianqin XU ; Jiaping SHENG ; Mingsun FANG ; Minli CHEN
Chinese Journal of Comparative Medicine 2015;(9):8-13,38
ObjectiveToinvestigatethekidneyyangdeficiencyconstitutionandhypothalamus-pituitary-adrenal ( HPA) axis dysfunction in Lewis rats .Method Two kidney yang deficiency models were established by subcutaneous injection of hydrocortisone and by adrenalectomy to induce hypothalamic -pituitary-adrenal ( HPA) axis dysfunction in Lewis rats.Wistar rats were used as control and compared with the two types of Lewis rat models of kidney yang deficiency .The degree of kidney yang deficiency of the Lewis rats was evaluated by examination and detection of the general signs , behavior , and neuroendocrine function , and so on .Result Compared with the normal Wistar rats , the body weight of Lewis rats was significantly higher than that of Wistar rats at the same age .The body temperature , urine volume and grip strength were significantly lower than those of Wistar rats ( P<0.01 for all ) .The memory ability of the Lewis rats was slightly decreased.The liver, kidney and adrenal indexes were significantly decreased in the Lewis rats (P<0.01).The levels of serum ACTH, CRH, cGMP, Cort and urine 17OHCS were significantly decreased (P<0.01).Conclusion A very slight deficiency of adrenal cortex function of Lewis rats is caused by genetic inheritance , and Lewis rats have the constitution of congenital kidney yang deficiency ( criticality or prophase of kidney yang deficiency ) .