2.Cloning of gdnf in the mouse testis and its expression in sertoli cells.
Cui-mi DUAN ; En-zhong LI ; Shi-qing ZHANG ; Chang-yong WANG ; De-xue LI
National Journal of Andrology 2007;13(11):975-978
OBJECTIVETo clone the glial cell line-derived neurotrophic factor (GDNF) from the mouse testis, construct the eukaryotic expression vector and transfect this vector into Sertoli cells in order to use the gdnf-transfected Sertoli cells as the feeder layer to cultivate spermatogonial stem cells (SSCs).
METHODSTotal RNA was extracted from the testes of normal mature mice and gdnf was cloned and amplified using RT-PCR, inserted into the eukaryotic expression vector and transfected into sertoli cells (TM4 cell line). Immunofluorescence with anti-GDNF antibodies was performed at 40 h following the transfection.
RESULTSgdnf cDNA was cloned successfully, and GDNF expressed after transfected into Sertoli cells.
CONCLUSIONThis study provides a basis for culturing SSCs with gdnf-transfected Sertoli cells as the feeder layer.
Animals ; Cloning, Molecular ; Gene Expression ; Glial Cell Line-Derived Neurotrophic Factor ; genetics ; Male ; Mice ; Mice, Inbred Strains ; RNA ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Sertoli Cells ; metabolism ; Testis ; cytology ; metabolism ; Transfection
3.Research and analysis of data source--analysis of the items of medical plants in Xiandai Bencao Gangmu.
Duan-yi LIN ; Chang-en ZHOU ; Xin-mei LAI ; Shu-jing YANG
China Journal of Chinese Materia Medica 2008;33(17):2094-2096
Scientific data is the source of innovation in knowledge. In order to change the situation that there is few information in plenty of data and to obtain useful knowledge which has high information content, it is necessary to clean data and ensure data's accuracy and without noise off when database is established initially. High-quality data comes from high-quality data source. But incomplete and incorrect and irregular data exist widely in the data source of Chinese materia medica. The phenomenon of synonyms and homonym is quite serious, and there is no unified description for the name and origin of Chinese materia medica among different data sources. So data processing including data analysis and research is very important in the establishment of Chinese materia medica database. In order to get the most accurate and standard data, this paper analyzed the items of Medical Plants in Xiandai Bencao Gangmu, including classification analysis of medical plants: distribution analysis of different classes and analysis of medical part; analysis of synonyms and homonym; analysis of incorrect data and analysis of advantage and disadvantage of data sources.
Materia Medica
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classification
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Plants, Medicinal
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classification
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Reference Books, Medical
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Terminology as Topic
4.Value of serum heart fatty acid binding protein in the assessment of cardiac function in patients with rheumatic heart disease
Chang-En DUAN ; Chao-Yuan ZHOU ; Xiao-Chen LIU ; Pei-Li HAN
Journal of Xinxiang Medical College 2018;35(2):125-128
Objective To investigate the value of the serum heart fatty acid binding protein (H-FABP) in the assessment of cardiac function in patients with rheumatic heart disease (RHD).Methods Forty-five RHD patients hospitalized in the First Affiliated Hospital of Xinxiang Medical University from January 2016 to December 2016 were selected as the observation group.In addition,45 subjects with normal cardiac function in the same period were selected as control group.The levels of serum H-FABP,B-type natriuretic peptide(BNP),left ventricular end-diastolic dimension(LVEDD),left ventricular end-systolic dimension(LVESD) and left ventricular ejection fraction(LVEF) were compared in the groups.The correlation between serum H-FABP and BNP,cardiac function classification was analyzed.Results The serum H-FABP and BNP levels and LVEDD,LVESD in the observation group were significantly higher than those in the control group(P < 0.05),but the LVEF in the observation group was significantly lower than that in the control group(P < 0.05).There were significant differences in the serum H-FABP,BNP levels and LVEDD,LVESD,LVEF among the patients with class Ⅱ,Ⅲ and Ⅳ of heart function (P <0.05).The serum H-FABP,BNP levels and LVEDD,LVESD in the patients with class Ⅲ and Ⅳ of heart function were significantly higher than those in the patients with class Ⅱ of heart function(P < 0.05);but the LVEF in the patients with class Ⅲand Ⅳ of heart function was significantly lower than that in the patients with class Ⅱ of heart function(P <0.05).Compared with the patients with class Ⅲ of heart function,the serum H-FABP,BNP levels and LVEDD,LVESD in the patients with class Ⅳ of heart function were significantly higher(P < 0.05);but the LVEF in the patients with class Ⅳ of heart function was significantly lower(P < 0.05).The serum H-FABP level was positively correlated with serum BNP level,LVEDD and LVESD in the observation group (r =0.579,0.743,0.740;P < 0.05).but it was negatively correlated with LVEF (r =-0.801,P <0.05).Conclusion The serum H-FABP level increases with the aggravation of heart failure in patients with RHD.It can be used as an important index to evaluate the heart function and prognosis of RHD patients.
5.Molecular biological study of glycoprotein IX gene defect in Bernard-Soulier syndrome.
Xiao-juan ZHAO ; Zhao-yue WANG ; Wei-ming DUAN ; Jian-xin FU ; Ming-en LU ; Jia-min WANG ; Xia BAI ; Chang-geng RUAN
Chinese Journal of Hematology 2003;24(9):480-483
OBJECTIVETo identify a mutation G2113-->A in the glycoprotein (GP)IX gene associated with Bernard-Soulier syndrome (BSS) and to investigate BSS pathogenesis.
METHODSAllele-specific restriction enzyme was used to analyze the samples of patient, her mother, her brother and 40 healthy volunteers. Site-directed mutagenesis was performed to construct a expression vector PD-IXG2113A harboring the mutation G2113-->A. Chinese hamster ovary (CHO) cells were transiently cotransfected with plasmids harboring the entire coding region of GPIbalpha, GPIbeta and GPIX or mutant GPIX, respectively. Expression of GPIbalpha and GPIX in transfected CHO cells were analysed with flow cytometer. GPIbalpha and GPIX in the cytoplasma of transfected CHO cells were analysed by immunostaining and Western blotting.
RESULTSThe patient was found to be homozygosity of the substitution, her mother and her brother be heterozygous. Expressions of GPIbalpha and GPIX in mutant CHO cells were remarkably reduced, but abundant in the cytoplasma.
CONCLUSIONThe mutation of Ala139(GCC)-->Thr(ACC) in the GPIX did not affect synthesis and assembly of GPIb/IX complex but influence its anchoring and expression on the cell surface, which was responsible for BSS.
Adult ; Animals ; Bernard-Soulier Syndrome ; genetics ; Blotting, Western ; CHO Cells ; Cricetinae ; Female ; Humans ; Mutation ; Platelet Glycoprotein GPIb-IX Complex ; genetics
6.17beta-estradiol stimulates proliferation of spermatogonia in experimental cryptorchid mice.
En-Zhong LI ; De-Xue LI ; Shi-Qing ZHANG ; Chang-Yong WANG ; Xue-Ming ZHANG ; Jing-Yan LU ; Cui-Mi DUAN ; Xiang-Zhong YANG ; Li-Xin FENG
Asian Journal of Andrology 2007;9(5):659-667
AIMTo investigate whether estrogen stimulates the proliferation of spermatogonia or induces spermatogenesis in cryptorchid mice.
METHODSMice were surgically rendered cryptorchid, then treated with different doses of 17beta-estradiol (E2) s.c. once a day. Mice were killed at sexual maturity (45 days of age), and histological analysis and immunofluorescence were performed. Serum follicle stimulating hormone (FSH), estradiol, testosterone and luteinizing hormone (LH) were measured.
RESULTSLow doses of E2 had no notable effect on spermatogonia, but at higher doses, E2 stimulated the proliferation of spermatogonia.
CONCLUSIONE2 has a dose-related mitogenic effect on spermatogonia.
Animals ; Cell Division ; drug effects ; Cryptorchidism ; physiopathology ; Disease Models, Animal ; Estradiol ; blood ; pharmacology ; Follicle Stimulating Hormone ; blood ; Luteinizing Hormone ; blood ; Male ; Mice ; Spermatogonia ; cytology ; drug effects ; pathology ; Testosterone ; blood
7.Xixin Decoction improves learning and memory ability of SAMP8 by enhancing neuroprotective effect and inhibiting neuroinflammation.
En-Long ZHAO ; Yong-Chang DIWU ; Hu ZHANG ; Li-Qi DUAN ; Xin-Yue HAN ; Ya-Li WANG ; Yuan ZHOU
China Journal of Chinese Materia Medica 2023;48(18):5032-5040
This study aimed to explore the possible effect of Xixin Decoction(XXD) on the learning and memory ability of Alzheimer's disease(AD) model senescence-accelerated mouse-prone 8(SAMP8) and the related mechanism in enhancing neuroprotective effect and reducing neuroinflammation. Forty SAMP8 were randomly divided into a model group(10 mL·kg~(-1)·d~(-1)), a probiotics group(0.39 g·kg~(-1)·d~(-1)), a high-dose group of XXD granules(H-XXD, 5.07 g·kg~(-1)·d~(-1)), a medium-dose group of XXD granules(M-XXD, 2.535 g·kg~(-1)·d~(-1)), and a low-dose group of XXD granules(L-XXD, 1.267 5 g·kg~(-1)·d~(-1)). Eight senescence-accelerated mouse-resistant 1(SAMR1) of the same age and strain were assigned to the control group(10 mL·kg~(-1)·d~(-1)). After ten weeks of intragastric administration, the Morris water maze was used to test the changes in spatial learning and memory ability of mice after treatment. Meanwhile, immunofluorescence staining was used to detect the positive expression of receptor for advanced glycation end products(AGER), Toll-like receptor 1(TLR1), and Toll-like receptor 2(TLR2) in the hippocampal CA1 region of mice. Western blot was employed to test the protein expression levels of silencing information regulator 2 related enzyme 1(SIRT1), AGER, TLR1, and TLR2 in the hippocampus of mice. Enzyme linked immunosorbent assay(ELISA) was applied to assess the levels of Aβ_(1-42) in the hippocampus of mice and the levels of nuclear factor κB p65(NF-κB p65), NOD-like receptor protein 3(NLRP3), tumor necrosis factor-α(TNF-α), and interleukin-1β(IL-1β) in the serum and hippocampus of mice. Compared with the model group, XXD significantly improved the spatial learning and memory ability of SAMP8, increased the expression of neuroprotective factors in the hippocampus, decreased the levels of neuroinflammatory factors, and inhibited the expression of Aβ_(1-42). In particular, H-XXD significantly increased the expression of SIRT1 in the hippocampus of mice, reduced the expression levels of NF-κB p65, NLRP3, TNF-α, and IL-1β in the serum and hippocampus of mice, and decreased the expression of AGER, TLR1, and TLR2 in the hippocampus of mice(P<0.05 or P<0.01). XXD may improve the spatial learning and memory ability of AD model SAMP8 by enhancing the neuroprotective effect and inhibiting neuroinflammation.
Humans
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Neuroprotective Agents/therapeutic use*
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Sirtuin 1/metabolism*
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Toll-Like Receptor 2/metabolism*
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NF-kappa B/metabolism*
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Tumor Necrosis Factor-alpha/metabolism*
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Neuroinflammatory Diseases
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NLR Family, Pyrin Domain-Containing 3 Protein/metabolism*
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Toll-Like Receptor 1/metabolism*
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Alzheimer Disease/genetics*
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Hippocampus