1.Effect of lipoxin A4 on lipopolysaccharide-induced oxidant stress in human renal tubular epithelial cells and possible mechanisms
Fen XU ; Hua HAO ; Hua DAI ; Lixiang LI ; Lei ZENG
The Journal of Practical Medicine 2017;33(1):51-55
Objective The study aimed to investigate the effect of lipoxin A4 (LXA4) on lipopolysaccharide (LPS)?induced oxidant stress in human renal tubular epithelial cells (HK2 cells) and possible underlying mecha?nisms. MethodsHK2 cells were divided into three groups: Control ,LPS and LPS+LXA4 groups. After cells were treated with indicated conditions,morphological changes were observed. The expressions of Nrf2 were detected by immunofluorescence and cells were collected for RT?PCR experiments.Results HK2 cells seemed disrupted and necrotic with the administration of LPS. However ,LXA4 could prevent cells from injury induced by LPS. LPS decreased Nrf2 expression and promoted it to translocate to cytoplasm ,while LXA4 could increase its expression and promote it to translocate to nucleus. Moreover ,LPS could decrease Nrf2 and its downstream molecule mRNA expressions,but LXA4 could reverse this effect. Conclusion Our results demonstrated that LXA4 effectively inhibit?ed HK2 cell oxidant stress via Nrf2 pathway.
2.BML-111, the analogue of lipoxin, inhibits Hela cell proliferation
Hua HAO ; Fen XU ; Liqing WU ; Xinxin ZHANG ; Hua DAI
The Journal of Practical Medicine 2014;(13):2045-2047
Objective To investigate the effect of BML-111 (the analogue of lipoxin) on uterine Hela cell (cervix cancer cell line) proliferation and the underlying mechanism. Methods Hela cells were stimulated by 50, 100, 200 and 400 μg/L BML-111, respectively, and cell viability was determined by MTT assay. Hela cells were divided into three groups:the control group (no treatment), the BML-111(200μg/L) group and the BML-111(200μg/L)plus Boc-2 (10μmol/L)group. Expression and location of P53 protein were detected by immunofluorescence. Expressions of NF-κB p65,P53 and CyclinD1 protein were detected by Western blotting. Results BML-111 (100, 200 and 400 μg/L) could effectively inhibit Hela cell viability compared with the control group (P < 0.05). P53 expression was shown decreased in both the nucleus and the cytoplasm without any change of P53 location , however, Boc-2 could reverse this effect. BML-111 could effectively inhibit P53 and CyclinD1 expression via NF-κB pathway and the effects could also be inhibited by Boc-2. Conclusions BML-111 can effectively inhibit Hela cell proliferation via FPR2 and NF-κB pathway.
4.Analysis of the factors effecting the expression efficiency of the green fluorescent protein gene in mouse embryonic stem cells
Hua YANG ; Jian-Xin DAI ; Xu-Ming DAI ; Ji-Liang FU
Academic Journal of Second Military Medical University 2001;22(4):319-321
Objective: To study the factors effecting the expression of the reporter green fluorescent protein (GFP) gene in the mouse embryo stem cell line R1. Methods: Three different kinds of GFP euko-expression vectors were constructed, and the expression efficiency was contrasted both at mRNA and protein levels after they were integrated into the chromosomes of host cells. Results: At protein level, the GFP expression level of the colonies transfected by the expression vector-pEF-GFP with the promoter of the peptide elongation factor (EF) were significantly higher than that of the colonies transfected by pCMV-GFP with CMV promoter and by pdCMV-GFP with double copies of CMV-GFP expression unit. There was no significant difference between the colonies transfected by pCMV-GFP and pdCMV-GFP. The detection results on mRNA level of GFP had the same tendency as that at protein level. Conclusion: (1) GFP gene expression efficiency controlled by EF promoter is distinctly higher than that by CMV promoter in NIH3T3 and R1 ES cell line.(2) A slight increase of the copy number of the foreign gene expression units in the host chromosome can not make obvious increase of its expression efficiency. (3) The vector express GFP in R1 ES cell line efficiently and stablely is obtained.
5.Analysis of the factors effecting the expression efficiency of the green fluorescent protein gene in mouse embryonic stem cells
Hua YANG ; Jian-Xin DAI ; Xu-Ming DAI ; Ji-Liang FU
Academic Journal of Second Military Medical University 2001;22(4):319-321
Objective: To study the factors effecting the expression of the reporter green fluorescent protein (GFP) gene in the mouse embryo stem cell line R1. Methods: Three different kinds of GFP euko-expression vectors were constructed, and the expression efficiency was contrasted both at mRNA and protein levels after they were integrated into the chromosomes of host cells. Results: At protein level, the GFP expression level of the colonies transfected by the expression vector-pEF-GFP with the promoter of the peptide elongation factor (EF) were significantly higher than that of the colonies transfected by pCMV-GFP with CMV promoter and by pdCMV-GFP with double copies of CMV-GFP expression unit. There was no significant difference between the colonies transfected by pCMV-GFP and pdCMV-GFP. The detection results on mRNA level of GFP had the same tendency as that at protein level. Conclusion: (1) GFP gene expression efficiency controlled by EF promoter is distinctly higher than that by CMV promoter in NIH3T3 and R1 ES cell line.(2) A slight increase of the copy number of the foreign gene expression units in the host chromosome can not make obvious increase of its expression efficiency. (3) The vector express GFP in R1 ES cell line efficiently and stablely is obtained.
6.STUDIES ON QUANTITY OF CERCARIAE SHEDDING FROM ARTIFICIAL INFECTED SNAILS
Wanquan HUA ; Jianrong DAI ; Yousheng LIANG ; Ming XU ; Yongliang XU ; Yuanding JIANG
Chinese Journal of Schistosomiasis Control 1992;0(06):-
Objective To explore the Schistosoma japonicum cercariae collected method and the quantity of cercariae obtained from a great quantity artificial infected snails. Methods In laboratory condition, Oncomelania snails were infected with schistosome miracidia. Sixty days post-infection all snails were divided into 7 shares. Cercariae shed from 1 share snails every day and the number of all shedding days were 40. Cercariae shed from snails were collected with low-velocity centrifuge and the cercariae sediment were weighted. Results One thousand and nine hundred g snails bred for 120 days post-infection, the infection and survival rates were 36. 00% and 51. 58%. Cercariae col-lected were 10. 5 g from 40 days collection. Cercariae quantity of shedding from 1 000 positive snails per day was 0. 257 4 g.
7.Effects of matrine on invasion and metastasis of leukemia cell line Jurkat.
Wei ZHANG ; Bi-Tao DAI ; You-Hua XU
Chinese Journal of Integrated Traditional and Western Medicine 2008;28(10):907-911
OBJECTIVETo study the inhibitory effects of matrine, in different concentrations, on invasion and metastasis of human acute lymphocytic leukemia cell line Jurkat.
METHODSIn vitro cultured Jurkat cells were treated by matrine in concentration of 0 g/L, 0.1 g/L, 0.15 g/L and 0.2 g/L, respectively. Then cell adhesion assay, cell migration assay and matrigel invasion assay were used respectively to observe the effects of matrine on adhesion, migration and invasive capacity of Jurkat cells. Meantime, RT-PCR was performed to detect the matrix metalloproteinase (MMP)-2 and MMP-9 mRNA expression levels. Comparison of measurement data among groups was analyzed by variance analysis.
RESULTSAs compared with the control group, the adhesion of Jurkat to fibronectin (FN) was significantly inhibited by 0.15 g/L and 0.2 g/L of matrine (P < 0.05); the cell migration and invasive capacity were significantly lowered by 0.1 g/L, 0.15 g/L and 0.2 g/L matrine (P < 0.01). High mRNA expression of MMP-9 presented but that of MMP-2 was expressed insignificantly in Jurkat cells, matrine at 0.1 g/L, 0.15 g/L and 0.2 g/L showed obvious effect in down-regulating MMP-9 mRNA expression (P < 0.01). Besides, MMP-9 mRNA expression was found to be positively correlated with the invasive capacity of Jurkat cells (r = 0.940, P < 0.01).
CONCLUSIONSMatrine is a good drug for antagonizing the invasion and metastasis of leukemia cells, it may roundly inhibit the adhesion, migration and invasive capacity of Jurkat cells, the mechanism might be related with the down-regulation of MMP-9 mRNA expression.
Alkaloids ; pharmacology ; Animals ; Cell Proliferation ; drug effects ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Jurkat Cells ; Leukemia ; drug therapy ; pathology ; physiopathology ; Mice ; NIH 3T3 Cells ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Quinolizines ; pharmacology ; Random Allocation
8.Discussion on effect of comprehensive community nursing on patients with primary hypertension
Junxiu WANG ; Hua XU ; Li DAI ; Rui DING ; Daowen YUAN ; Guofang ZHANG
Chinese Journal of Practical Nursing 2009;25(9):1-3
Objective The optimal community nursing intervention methods for community patients with primary hypertension was discussed in order to promote their health. Methods System management and community intervention were applied to 140 community patients with primary hypertension for 2 years, including strengthing community communication, popularizing health knowledge, improving cognition of danger of hypertension, hypertensive disease project management, tracking and monitoring the patients with primary hypertension and medication supervision.The cognition rate of prevention and control knowledge for hypertension, hypertension-related cardiovascular complication and formation rote of hypertension control behavior were compared before and after intervention. Results The prevalence rate of the knowledge on hypertension was increased, accompanied with promoting regular medication in patients with hypertension, decreasing cardiovascular complication caused by hypertension and heightening formation rate of controlling behavior in hypertension patients.Conclusions Applying community nursing intervention is practically effective in prevention and treatment of primary hypertension, deserving promotion and application in community medical institute.
9.Use of antihypertensive drugs among community hypertensive patients in Shanghai
Jingli WANG ; Xianggui XU ; Jingzhen WANG ; Ning MO ; Yaocheng ZHU ; Junling GAO ; Junming DAI ; Hua FU
Chinese Journal of General Practitioners 2010;09(9):603-606
Objective To investigate use of oral antihypertensive drugs among community hypertensive patients in Shanghai and find out factors related to their unreasonable use to direct their clinical use. Methods Seven hundred and three hypertensive patients were surveyed with questionnaire by stratified cluster sampling at three neighborhoods and one village of Dahua community, Baoshan district, Shanghai during April to June 2009 to understand their use of antihypertensive agents, including kinds and forms of drugs, rationale of drug use. Results Five hundred and eighty-two (82. 8% ) of 703 hypertensive patients interviewed were using antihypertensive drugs, 271 (38. 5% ) of them used only one kind of non-compound antihypertensive drug, 182 (25.9%) used one kind of compound agent, 311 (53.4%) used two or more kinds of drugs in combination, including 117 patients ( 16. 8% ) used two kinds of agents combined and 12 patients ( 1.6% ) used three kinds of agents combined. Two hundred and sixty-six (47.5%) patients took orally calcium-channel blockers and 205 ( 35.2% ) used compound agents. Conclusions Frequency of combined use of two or more kinds of antihypertensive agents is reasonable and significantly higher than that of use of one kind of drug at Dahua community in Shanghai. Calcium channel blocker plays a predominant role in treatment for hypertension, non-long-acting compound agents are used in a higher proportion. But,guidelines for hypertension prevention and treatment are not so well complied with in local hypertensive patients. So, it is suggested that training for community physicians and management for standard use of antihypertensive agents at community should be strengthened further.
10.Type 2 epithelial mesenchymal transition in vivo: truth or pitfalls?
Chinese Medical Journal 2012;125(18):3312-3317
Epithelial-mesenchymal transition (EMT) is a process by which fully differentiated epithelial cells undergo a phenotypic conversion and assume a mesenchymal cell phenotype, including elongated morphology, enhanced migratory and invasiveness capacity, and greatly increased production of extracellular matrix (ECM) components. The EMTs associated with wound healing, tissue regeneration, and organ fibrosis are termed as type 2 EMT. Over the past two decades, emerging evidence suggested that injured epithelial cells, via type 2 EMT, may serve as important sources of fibroblasts and contribute to organ fibrosis, such as kidney, liver, lung and eyes. There is perhaps no doubt that adult epithelial cells can undergo EMT in vitro in response to transforming growth factor (TGF)-β1 and other inflammatory or pro-fibrotic stimuli. However, whether type 2 EMT really occurs in vivo, whethers it is actually a source of functional and activated interstitial fibroblasts and whether it contributes to tissue fibrosis have already been the subjects of heated debate. In this review, we will describe the main features of EMT, the major findings of type 2 EMT in vitro, the evidences for and against type 2 EMT in vivo and discuss the heterogeneity and pitfalls of the techniques used to detect EMT during fibrotic diseases. We suggest that in order to ascertain the existence of type 2 EMT in vivo, different proper phenotype markers of epithelial and mesenchymal cells should be jointly used and cell lineage tracking techniques should be standardized and avoid false positives. Finally, we believe that if EMT really occurs and contributes to tissue fibrosis, efforts should be made to block or reverse EMT to attenuate fibrotic process.
Animals
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Epithelial-Mesenchymal Transition
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physiology
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Fibroblasts
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cytology
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metabolism
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Fibrosis
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metabolism
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pathology
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Humans