1.Construction of basic fibrobiast growth factor adenovinis vector and its expression in human umbilical vein endothelial cells
Lingli GUO ; Xin XING ; Kan LIU ; Dali HAN
Academic Journal of Second Military Medical University 1981;0(04):-
Objective:To construct an adenovirus vector harboring human basic fibroblast growth factor (bFGF) cDNA and investigate the expression of bFGF in human umbilical vein endothelial cells (HUVEC) in vitro. Methods: The adenovirus expression vector Ad5-bFGF was constructed by homologous recombination technique. The best value of MOI was tested by transfecting human umbilical vein endothelial cells (HUVEC) with Ad5-GFP. Ad5-bFGF was used to transfect HUVEC at the obtained value of MOI and the expression of bFGF protein was detected by immunocytochemistry method and Western blotting. Results: The best value of MOI for adenovirus 5 to transfect HUVEC was 200 and the transfection rate was 90%. Immunocytochemistry method and Western blotting showed that bFGF was expressed in HUVEC after transfection with Ad5-bFGF and the expression was significantly higher than that in untransfected HUVEC (P
2.The mechanism of electroacupuncture for regulating gastrointestinal motility in rabbits
Guo-Dong HE ; Wei-Xin NIU ; Han LIU ; Al ET ;
Chinese Journal of Digestion 2001;0(11):-
Objective To investigate the mechanism of regulating gastrointestinal motility by elec- troacupunturing point of"Zusanli"in rabbits.Methods Thirty adult rabbits were divided into control, electrical acupuncture point of"Zusanli"and non acupuncture point groups.The blood concentrations of motilin were detected at different times(before acupuncture and 15,30,45 and 60 min after acupunc- ture).Sixty min after acupuncture,the mice were sacrificed.The acetylcholine esterase and nitric oxide synthase(NOS)in gastric and jejunum tissues were examined.The electron microscope was used to observe the vesicles of nerve ending.Results The concentrations of motilin in electrical acupuncture group was significantly increased and reached the high peak at 45min.The activities of acetylcholine es- terase was increased significantly in electrical acupuncture group compared to control group[gastric tis- sue:(15 571?2876)pinel vs(9081?801)pinel,P
3.Observations on the Therapeutic Effect of Heat-sensitive Point Thunder-fire Moxibustion on Knee Osteoarthritis
Qingdong YUAN ; Xin GUO ; Yacen HAN ; Jingqian ZHANG ; Xiaodong FENG
Shanghai Journal of Acupuncture and Moxibustion 2015;(7):665-668
Objective To investigate the clinical efficacy of heat-sensitive point thunder-fire moxibustion in treating knee osteoarthritis (KOA). Methods One hundred and forty-eight KOA patients were randomly allocated to treatment and control groups, 74 cases each. The treatment group received heat-sensitive point thunder-fire moxibustion and the control group took diclofenac sodium enteric-coated tablets. The Visual Analogue Scale (VAS) score, the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score and 50 yards fastest walking time were observed in the two groups before and after 30 days of treatment. The clinical therapeutic effects were compared between the two groups. Results There were statistically significant pre-/post-treatment differences in the VAS score and WOMAC subscores in the two groups (P<0.01). There was a statistically significant pre-/post-treatment difference in 50 yards fastest walking time in the treatment group (P<0.05). There were statistically significant post-treatment differences in the VAS score, the WOMAC score and the WOMAC pain and stiffness scores between the treatment and control groups (P<0.01). There were statistically significant differences in the VAS and WOMAC scores at three months after treatment between the treatment and control groups (P<0.01). The total efficacy rate was 95.9%at the end of treatment and 95.6%at three months after treatment in the treatment group, and 86.1%at the end of treatment and 86.8%at three months after treatment in the control group; there were statistically significant differences between the two groups (P<0.05). Conclusion Heat-sensitive point thunder-fire moxibustion is an effective way to treat knee osteoarthritis.
4.miRNA-101 inhibits the expression of the enhancer of zeste homolog 2 in androgen-independent prostate cancer LNCaP cell line.
Jian-xin LIU ; Qi-fa ZHANG ; Chang-hai TIAN ; Yong ZHANG ; Xiao-zhou HAN ; Hao GUO
National Journal of Andrology 2015;21(6):500-503
OBJECTIVETo investigate the effect of miRNA-101 on the expression of the enhancer of zeste homolog 2 (EXH2) in human androgen-independent prostated cancer LNCaP cells.
METHODSWe divided LNCaP cells into a blank control, a negative control, and a miRNA-l01 transfection group, constructed the vector by transfecting synthetic miRNA-101 mimics into the LNCaP cells, and evaluated the efficiency of transfection by fluorescence microscopy. Then we determined the expression level of EZH2 mRNA by qRT-PCR in the three groups of cells and that of the EZH2 protein in the negative control and transfection groups by Western blot.
RESULTSGreen fluorescence signals were observed in over 70% of the LNCaP cells in the transfection group after 24 hours of transfection. At 72 hours, the expression of miRNA-101 was significantly upregulated in the transfected cells (P < 0.01), that of EZH2 mRNA was remarkably lower in the transfection group (0.01 ± 0.10) than in the blank control (0.95 ± 0.40) and negative control (0.86 ± 0.30) groups (both P < 0.01), and that of the EZH2 protein was increased in the negative control but decreased in the transfection group with the extension of culture time.
CONCLUSIONmiRNA-101, with its inhibitory effect on the expression of EZH2 in LNCaP cells, is a potential biotherapeutic for prostate cancer.
Androgens ; Cell Line, Tumor ; Enhancer of Zeste Homolog 2 Protein ; Genetic Vectors ; Humans ; Male ; MicroRNAs ; physiology ; Polycomb Repressive Complex 2 ; genetics ; metabolism ; Prostatic Neoplasms ; metabolism ; RNA, Messenger ; metabolism ; Transfection
5.Detection and significance of fusion gene between TMPRSS2 and ETS transcription factor genes in fresh prostatic cancer tissues in Chinese patients.
Hua XIANG ; Zong-xin LING ; Ke SUN ; Guo-ping REN ; Qi-han YOU ; Xiong-zeng ZHU
Chinese Journal of Pathology 2011;40(3):187-188
Carcinoma
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genetics
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metabolism
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pathology
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surgery
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China
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Humans
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Male
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Oncogene Proteins, Fusion
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genetics
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Prostatic Hyperplasia
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genetics
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metabolism
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pathology
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surgery
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Prostatic Neoplasms
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genetics
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metabolism
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pathology
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surgery
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Proto-Oncogene Proteins c-ets
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genetics
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metabolism
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Reverse Transcriptase Polymerase Chain Reaction
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Serine Endopeptidases
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genetics
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metabolism
6.The effective parts of liangxue tongyu prescription on cooling-blood and activating-blood and analysis of chemical constituents by HPLC-MS and GC-MS.
Xi HUANG ; Guo-chun LI ; Lian YIN ; Zi-han ZHANG ; Yi-xin LIANG ; Hai-bo CHEN
Acta Pharmaceutica Sinica 2015;50(1):86-93
In order to clarify material basis of effective parts of liangxue tongyu prescription, blood-heat and blood-stasis rat model induced by dry yeast was established. The changes of rectal temperature, blood viscosity and plasma viscosity were used to evaluate the cooling-blood and activating-blood effects of liangxue tongyu prescription and its parts. Compared with the model group, the extract from liangxue tongyu prescription, its volatile oil and n-butanol part could significantly reduce rectal temperature (P<0.01), and also reduce blood viscosity and plasma viscosity to various degrees (P<0.01 or P<0.05). So volatile oil and n-butanol part were primarily identified as effective parts of liangxue tongyu prescription. By using GC-MS with normalization method of area to analyze volatile oil of liangxue tongyu prescription, 70 compounds were identified, accounting for about 92.54%, mainly as β-asarone, paeonol, α-asarone and shyobunone. 42 compounds such as peony glycosides, tannins, and iridoid glycosides were identified by HPLC-MS techniques and standard comparison. The study determined the effective parts of liangxue tongyu prescription and clarified the chemical composition providing the foundation for further studies on material basis of liangxue tongyu prescription.
Acetophenones
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chemistry
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Animals
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Anisoles
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chemistry
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Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal
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chemistry
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Gas Chromatography-Mass Spectrometry
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Oils, Volatile
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chemistry
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Rats
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Tannins
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chemistry
8.Dimethyl sulfide, a metabolite of the marine microorganism, protects SH-SY5Y cells against 6-hydroxydopamine and MPP+-induced apoptosis
WU PENG-FEI ; GUAN XIN-LEI ; LUO HAN ; WANG FANG ; CHEN JIAN-GUO
Chinese Journal of Pharmacology and Toxicology 2017;31(10):1004-1004
Dimethyl sulfide (DMS) has been historically recognized as a metabolite of the marine microorganism or a disgusting component for the smell of halitosis patients. In our recent study, DMS has been identified as a cytoprotectant that protects against oxidative-stress induced cell death and aging. We found that at near- physiological concentrations, DMS reduced reactive oxygen species (ROS) in cultured PC12 cells and alleviated oxidative stress. The radical-scavenging capacity of DMS at near-physiological concentration was equivalent to endogenous methionine(Met)-centered antioxidant defense. Methionine sulfoxidereductase A (MsrA), the key antioxidant enzyme in Met-centered defense, bound to DMS and promoted its antioxidant capacity via facilitating the reaction of DMS with ROS through a sulfonium intermediate at residues Cys72, Tyr103, Glu115, followed by the release of dimethyl sulfoxide (DMSO). MTT assay and trypan blue test indicated that supplement of DMS exhibited cytopro?tection against 6-hydroxydopamine and MPP + induced cell apoptosis. Furthermore, MsrA knockdown abolished the cytoprotective effect of DMS at near- physiological concentrations. The present study reveals new insight into the potential therapeutic value of DMS in Parkinson disease.
9.Effects of Sappan Lignum and Chuanxiong Rhizoma on Expression of ABCG2 Protein of PG-BE1 Stem Like Cells in Vivo
Yaohan WANG ; Peitong ZHANG ; Dong YANG ; Haiying HAN ; Xiuwei GUO ; Xin QI ; Yun ZHANG
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(8):60-65
Objective To observe the effects of different doses of Sappan Lignum and Chuanxiong Rhizoma on tumor stem cells marker ABCG2 in vivo. Methods Sphere cells obtained from serum culture were inoculated in nude mouse armpit, which were randomly divided into 5 groups: control group, Sappan Lignum high- and low-dose groups, and Chuanxiong Rhizoma high- and low-dose groups. Each medication group was given relevant medicine for gavage. 21 days later, inhibition tumor rate and ABCG2 protein and mRNA expression were detected with confocal microscope, Western blot, and RT-PCR. Results The sphere cells obtained from serum free culture had the abilities of cancer stem cells, such as proliferation, anti-aptosis and high expression of cancer stem cells markers. Chuanxiong Rhizoma high- and low-dose groups could inhibit tumor growth (P<0.05), and the inhibitory rate of Chuanxiong Rhizoma low-dose group was higher than the Chuanxiong Rhizoma high-dose group. Sappan Lignum high- and low-dose groups inhibited tumor growth without statistical significiance (P>0.05). Compared with the control group, Chuanxiong Rhizoma low-dose group could significantly inhibit the expression of resistant protein of ABCG2. Sappan Lignum high- and low-dose groups could not inhibit the protein expression of ABCG2. Each medication group up-regulated the mRNA expression of ABCG2 except for Chuanxiong Rhizoma low-dose group. Conclusion Low dose of Chuanxiong Rhizoma can inhibit the expression of ABCG2 protein levels, which can be the targeting killer for cancer stem cells.
10.Free pharmaceutical policy to promote equity and access:Its implications for China
Xiaodong GUAN ; Baomin WANG ; Xiaoxiong XIN ; Zhigang GUO ; Sheng HAN ; Luwen SHI
Chinese Journal of Health Policy 2015;(2):54-59
Promoting equity and access to medicine is important to guarantee health equity. The international communities commonly guarantee equity and access to medicine through free pharmaceutical policy. In most coun-tries, free pharmaceutical policy covers patients who cannot afford, children under 5, pregnant women and elderly people. The free medicines include essential medicines, medicines for chronic and infectious diseases, vaccines, etc. Financing of free medicines comes from health insurance, direct government investment and international aid, and the medicine delivery mainly depends on the public medical institutions. Free pharmaceutical policy in China mainly refers to medicines for infectious diseases and vaccines, and the coverage is narrow. This paper argues that since China has been becoming one of the middle income countries, the amount of free medicines should be in-creased, especially for chronic diseases, such as diabetes, hypertension, etc. The equity and access to medicine in China should be further promoted by providing all essential medicines for free.