3.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.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.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
8.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.Analysis of Prognositic Factors of Severe Traumatic Brain Injury in Pediatric Intensive Care Unit
hai-rong, GONG ; zhu-jin, LU ; guo-ping, LU ; ling-en, ZHANG
Journal of Applied Clinical Pediatrics 1994;0(04):-
5 days.Blood gas analysis and blood pressure were determined at admitted day.Meanwhile,peripheral white blood cells at d1,3,5,and blood glucose were measured every day,respectively.GCS at d1,3,5 and hyperglycemia scorce(HS) were evaluated.Results Of the 82 studied patients,36 cases died.Univariate analysis showed that hypotension,lower GCS,higher peripheral white blood cells and HS were the independent death risk factors(Pa0.05).In multivariate logistic regression,the factors significantly associated with an increase in mortality were hypotension,lower GCS and higher HS.Conclusion Lower GCS,higher HS and hypotension are associated with poor outcome of children with severe trauma brain injury.
10.Processing Technology Research of Fermentation and Purification of SUMO Protease UlP1
Xiu-Ping FENG ; Bai-Rong DU ; Dong-Mei YAN ; Xiang-Feng ZHAO ; Xun ZHU ;
China Biotechnology 2006;0(02):-
Nowadays,small peptides are always expressed in the form of fusion protein.The expression product contains many superfluous amino acids which can affect the biological functions of small peptides even expressed by GST fusion protein expression system.SUMO protease can cut SUMO fusion protein expressed by fusion expression system without any amino acid residues left on target protein thus become a hot topic in this field.Recombinant His-UlP1/pET3c/BL21(DE3)engineering strain was constructed by genetic engineering technology and the expression conditions were optimized in shake flaks.The process of high density fermentation was explored and different purification conditions were detected by chromatography.The results showed that SUMO protease could be expressed well after inducing the engineering strain by IPTG of 1.0mmol/L at 30℃ for 6 hours.The expression level of the strain in fermentation pots could reach 24.3% analyzed by SDS-PAGE.The purity of SUMO protease was more than 98% after further purification by cation exchange chromatography.The yield was 355mg SUMO protease per liter fermentation liquid.Western blot analysis demonstrated that there were immune reactions between IlP1 and 6?His antibodies,so it has established a good foundation for large-scale industralazation in the future.