2.Overexpression of caveolin-1 inhibits the growth of human cervical squamous cell Hela cell line in vitro
Qingling ZHENG ; Donghua GU ; Jinliang PING ; Rong ZHU ; Qi CHEN
Journal of Chinese Physician 2010;12(10):1304-1308
Objective To investigate the effects of caveolin-1 overexpressing on the growth of cervical squamous cell cancer Hela cell line. Methods Eukaryotic expression vector of human caveolin-1 gene was introduced into Hela cells by Lipofectamine. The clones stably overexpressed caveolin-1 were identiffed by RT-PCR, immunofluorescence cell staining techniques and Westernblotting. Cells proliferation viabihty was tested by MTT assay, and flow cytometry was used to assay the cell cycle and apoptosis, and the relative phosphorylation level of extracellular regulated protein kinases (Erk1/2) were detected by Westernblotting. Results The clones stably overexpressed caveolin-1 were obtained. Compared with the parental Hela cells, the tranfected cells exhibited a slower rate of growth. FAGS analysis results revealed that overexpression of caveolin-1 resulted in the cell cycle arrest in the G0/G1 [ ( 68. 04 ± 2. 57 ) % vs ( 53.41 ±1.01)%] phase and increased the apoptotic cell fraction[ (19. 18 ±2.20)% vs (5.63 ±0.55)%, P <0. 05 ]. Western blotting results showed that overexpression of caveolin-1 reduced the phosphorylation of Erk1/2(0.28 ±0.05 vs 0.81 ±0.07, P <0.05). Conclusions Overexpression of caveolin-1 suppressed the growth of Hela cells and induced apoptosis, down-regulation of Erk1/2 phosphorylation might be involved in its mechanism.
5.Complete genome sequence analysis of Japanese encephalitis virus newly isolated in China.
Rong-Hui XIE ; Han-Ping ZHU ; Shi-Hong FU ; Yin-Kai CHENG ; Fang XU ; Ping-Ping YAO ; Zhang-Nv YANG ; Xiao-Long ZHOU ; Zhi-Yong ZHU
Chinese Journal of Experimental and Clinical Virology 2009;23(4):245-247
OBJECTIVETo study the complete genome sequence of Japanese encephalitis virus (JEV) strain XJ69 isolated in ZheJiang province and explore its evolution.
METHODSOverlapping primers were designed according to the full-length genomes from GenBank. RT-PCR was used to amplify the fragments and RT-PCR products were cloned T vector, sequenced and analyzed.
RESULTSThe genome of strain XJ69 and XJP613 were 10 964 nucleotides in length with a single open reading frame encoding 3432 amino acids. Comparison of the complete genome sequences of different JEV isolates showed XJ69 and XJP613 were 83.5%-99.2% and 83.4%-99.4% nucleotide sequence homology among them respectively, which resulted in 94.8%-99.7% amino acid sequence homology. Phylogenetic analysis through PrM/C,E and full-length genome showed that the XJ69 and XJP613 strain belonged to genotype I.
CONCLUSIONThe nucleotitede sequence and deduced amino acid sequence of XJ69 and XJP613 strain were similar to that of those of genotype I of Japanese encephalitis virus. It belonged to genotype I and were close to the isolates SH17M-07.
Animals ; Cell Line ; China ; Cricetinae ; Encephalitis Virus, Japanese ; classification ; genetics ; isolation & purification ; Encephalitis, Japanese ; virology ; Genome, Viral ; Humans ; Molecular Sequence Data ; Phylogeny
6.Effect of Bushen Tongdu Capsule on RANK/RANKL/OPG pathway of collagen induced arthritis rats.
Yang-Chun ZHU ; Lin LIN ; Xiao-Li ZHOU ; Rong-Fang LI ; Li-Ping HOU
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(12):1487-1491
OBJECTIVETo study the effect of Bushen Tongdu Capsule (BTC) on RANK/RANKL/ OPG pathway of collagen induced arthritis (CIA) rats, thereby laying theoretic evidence for treating rheumatic arthritis (RA) by Chinese medicine.
METHODSRA model was induced by CIA. Totally 42 rats were randomly divided into six groups, i.e., the normal control group, the model group, the low dose BTC (BSL) group, the medium dose BTC (BSM) group, the high dose BTC (BSH) group, and the Tripterygium Glycosides (TG) group, 7 in each group. BTC at the daily dose of 120, 240, and 480 mg/kg was given by gastrogavage to rats in the BSL, BSM, and BSH group respectively from the 13th day of modeling. TG at the daily dose of 24 mg/kg was given by gastrogavage to rats in the TG group. All medication was given once daily, 2 mL each time. Two mL normal saline was administered to rats in the normal control group and the model group. All medication lasted for 18 days. Samples were taken at day 31. The TRAP section of the ankle joint was fixed in 10% formalin for TRAP stain. Serum levels of osteoprotegerin (OPG), receptor activator of nuclear factor-κB ligand (RANKL), and macrophage colony-stimulating factor (M-CSF) were detected using ELISA.
RESULTSCompared with the normal control group, positive reactions of pathological ankle joint section, inflammation, and osteoclasia degree were significantly improved in the model group, serum levels of RANKL and M-CSF were up-regulated, levels of OPG and OPG/RANKL were significantly lowered (all P < 0.01). Compared with the model group, positive reactions of pathological ankle joint section, inflammation, and osteoclasia degree also significantly decreased in the BSH group and the TG group (all P < 0.01). RANKL and M-CSF were significantly down-regulated in each medicated group, while levels of OPG and OPG/RANKL were significantly up-regulated (all P < 0.01). Compared with the TG group, M-CSF was lower, but levels of OPG and OPG/RANKL were significantly up-regulated in the normal control group (all P < 0.01). RANKL and M-CSF were significantly up-regulated, while levels of OPG and OPG/RANKL were significantly down-regulated in the model group and each BS group (all P < 0.01).
CONCLUSIONBTC could relieve bone damage of CIA rats possibly through regulating and controlling osteoclasts.
Animals ; Arthritis, Experimental ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Inflammation ; Macrophage Colony-Stimulating Factor ; metabolism ; Osteoclasts ; Osteoprotegerin ; metabolism ; RANK Ligand ; metabolism ; Rats ; Receptor Activator of Nuclear Factor-kappa B ; metabolism ; Tripterygium
7.Outcome analysis of endemic fluorosis control in Guangxi in 2006
Min, LIAO ; Jun, LIU ; Zhen-zhu, TANG ; Li-rong, HUANG ; Rui-jun, NING ; Xuan-ping, ZENG
Chinese Journal of Endemiology 2008;27(3):300-302
Objective To investigate the distribution of water fluoride and the status of water-improving defluoridation projects,thus to explore the condition of endemic fluorosis in Guangxi Province.Methods According to"The National Technical Scheme for Endemic Disease Contml in 2005",the fuorine content in water Was determined by F-ion selective electrode,children's dental fluorosis was checked by Dean method.and the skeletal fluorosis was checked by the standard of clinical scale of skeletal fluorosis.Results 305 water samples in 61 villages were examined,among which 71 waters were exceeded the standard,accounting for 23.28%(71/305).The projects of defluoriding drinking water were running well except one was discarded.The prevalence of dental fluorosis was 13.55%(356/2627),the prevalence of skeletal fluorosis was 4.02%(65/1615).Conclusions The situation of endemic fluorosis control is not optimistic in Guangxi,which needs fuaher prevention and controls.
9.Effect of yigu capsule contained serum on mRNA and protein expression of estrogen receptor in osteoblast rats.
Rong-Hua ZHANG ; Ke-Ping PENG ; Xiao-Feng ZHU
Chinese Journal of Integrated Traditional and Western Medicine 2005;25(4):333-337
OBJECTIVETo explore the effect of Yigu capsule contained serum on mRNA and protein expression of osteoblast estrogen receptor (ER), for investigating the mechanism of its preventing and treating osteoporosis by means of regulating estrogen.
METHODSOsteoblasts separated from newborn SD rats were cultured, and divided into 3 groups after being passaged, i.e. the drug-serum treated group, the blank serum group and the DMEM medium control group. The relative amount of ERalpha and ERbeta expression were determined with RT-PCR, the affinity (expressed by equilibrium dissociation constant, KD) and number of ER (RT) were analysed by radioligand assay.
RESULTSThe relative amount of ERbeta mRNA expression were increased in the drug-serum group, with significant difference as compared with that in the other two groups (P < 0.05), but no significant difference among the three groups in ERa mRNA expression (P > 0.05). KD in the drug-serum group showed insignificant difference as compared with that in the other two groups (P > 0.05), but RT increased in the drug-serum group and the difference in the three groups was significant (P < 0.01).
CONCLUSIONDrug-contained serum of Yigu capsule can up-regulate the expression of osteoblast ERbeta mRNA and increase the amount of ERs. Regulating estrogen is possibly one of the mechanisms of Yigu capsule in preventing and treating osteoporosis.
Animals ; Capsules ; Cell Proliferation ; drug effects ; Cell Separation ; Drugs, Chinese Herbal ; pharmacology ; Female ; Osteoblasts ; cytology ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Receptors, Estrogen ; biosynthesis ; genetics ; Serum
10.Thri-operators and the blood-oxygen functional image technology in diagnosing breast cancer
Jianhua SHI ; Linlin ZHEN ; Yumei ZHOU ; Zhaohuan ZHU ; Rong WU ; Xuedong HAN ; Ping ZHANG
Journal of Endocrine Surgery 2009;3(3):161-164
Objective To evaluate thri-operators and the blood-oxygen functional image technology in di-agnosing breast cancer. Methods One hundred and forty-six patients were admitted to hospital for operation due to one hundred and fifty-three suspicious lesions detected in their breasts. These lesions were detected by physical examination, thri-operators and the blood-oxygen functional image, mammography uhrasonography. The sensitivity and specificity of each diagnostic method were obtained and the radiolagie-pathologic correlation was meanwhile calculated. Results Sixty one(41.8%)breast lesions were diagnosed as malignancy. The sensitivity, specificity, accuracy,positive prognostic value and negative prognostic value of ultras onography were 80. 33% ,89. 41%,85.61% ,84.48% and 86.36%. Such data of mammography were 57.89% ,80. 36% ,69.03% ,75.00% and 65. 22%. And those of thri-operators and the blood-oxygen functional image technology were 91.80% ,83.53%, 86.99% ,80.00% and 94.67%. Conclusions Thri-operators and the blood-oxygen functional image technology is superior to uhrasonography and mammography in diagnosing breast lesions with its sensitivity accuracy and neg-ative prognostic value, while specificity and positive prognostic value were between them, have greater value in screeninging and the diagnosing breast cancer.