1.Basic fibroblast growth factor promotes the differentiation of rat bone marrow mesenchymal stem cells into Leydig cells.
Xiu-wen YAN ; Chun LIU ; Fei TIAN
National Journal of Andrology 2015;21(6):494-499
OBJECTIVETo explore the role of the basic fibroblast growth factor (bFGF) in the directional differentiation of bone marrow mesenchymal stem cells (BMSCs) into Leydig cells.
METHODSAfter purification and identification, we inoculated the third-generation BMSCs of SD rats onto a six-orifice board and then randomly divided them into groups A (normal saline control), B (human chorionic gonadotropin [hCG] + platelet-derived growth factor [PDGF] induction), C (hCG + PDGF + 5.0 ng/ml bFGF induction), D (hCG + PDGF + 10.0 ng/ml bFGF induction), and E (hCG + PDGF + 20.0 ng/ml bFGF induction). On the 7th, 14th and 21st day of induction, we observed the morphological changes of the cells and measured the level of testosterone (T) and expression of 3 beta hydroxy steroid dehydrogenase (3β-HSD) in the supernatant by immunofluorescence staining.
RESULTSAfter induction, the BMSCs of groups B, C, D, and E exhibited microscopic features of enlarged size, inter-connection, long-shuttle or irregular shape, adherent growth, and large round nuclei, all characteristic of Leydig cells. With the prolonging of time and enhanced concentration of bFGF, gradual increases were observed in the T level and the count of 3β-HSD-positive BMSCs in the four induction groups, with statistically significant differences between group B and groups C, D, and E (P < 0.05), as well as between group C and groups D and E (P < 0.05), but not between D and E (P > 0.05).
CONCLUSIONThe bFGF has an obvious promoting effect in the in vitro induced differentiation of rat BMSCs into Leydig cells.
Animals ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Differentiation ; Cells, Cultured ; Chorionic Gonadotropin ; metabolism ; Fibroblast Growth Factor 2 ; pharmacology ; Humans ; Leydig Cells ; cytology ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Testosterone ; metabolism
2.Establishment of Rat Models with Alzheimer's Disease Induced by ?-Amyloid Protein and Effects of Nicotine on the Animal Models
yong-bo, ZHAO ; wen-wen, LIU ; chun-ni, GUO
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(07):-
Objective To establish rat models with Alzheimer's disease(AD) induced by ?-amyloid,and to observe the effects of nicotine on learning and memorizing ability of the rats. Methods Aggregated A?1-40 was injected into the bilateral basal forebrain of the rats.Learning and memorizing ability of the rats were inspected through Morris water maze test.The effects of nicotine on learning and memorizing ability of the rats induced by ?-amyloid peptide were observed. Results The learning and memorizing ability of AD animal models injected aggregated(A?1-40) for 2 weeks decreased(P
3.Expression of Notch2,Notch4 in Hyperoxia Induced Lung Injury in Premature Rats
chun-mei, LIU ; li-wen, CHANG ; jing, LIU
Journal of Applied Clinical Pediatrics 1986;0(02):-
Objective To explore the roles of Notch2,Notch4 in hyperoxia induced lung injury in premature rats.Methods At the postnatal 1 day Sprague-Dawley premature rats were randomly assigned to about 85% hyperoxia group and air group.At the 1,7,14,21 days after exposed,8 rats of each group were used to evaluate expressions of Notch2,Notch4 in lungs by immunohistochemistry and the level of Nothch2,Notch4 mRNA by reverse transcription polymerase chain reaction(RT-PCR).Results Expressions of Notch2,Notch4 had their rules in rats′ different stages of development;85% oxygen exposed would change their tracks.Conclusion Prolonged 85% oxygen exposure result in abnormal expressions of Notch2 and Notch4 ,which is likely to lead to the pathogenesis of hyperoxic lung injure in premature rats.
4.Transinfection of rabbit chondrocytes via chitosan microsphere as gene carriers with recombined human IL-1Ra gene and TGF-β1 gene
Xiaoyue WEN ; Daozhang CAI ; Huading LU ; Chun ZENG ; Bin LIU
Chinese Journal of Microsurgery 2009;32(6):471-473,封3
Objective To explore transinfection of rabbit chondrocytes via chitosan microsphere with human IL-1Ra and TGF-β1 gene. Methods Chitosan-DNA microspheres carrying plasmids with IL-1 Ra and TGF-β1 genes were prepared.The encapsulation efficiency,DNA-released kinetics and lysozyme degradation in vitro were performed.Articular rabbit chondrocytes were co-transinfected with the plasmids with IL-1Ra and TGF-β1 genes via chitosan-DNA mierosphere,evaluated by fluorescence microscope,TaqMan real-time PCR assay and MTF test. Results The chitosan microspheres with IL-1Ra and TGF-β1 genes were(2.8±0.2)μm and(2.6±0.1)μm in diameters respectively.The encapsulation efficiency were(88.3±4.1)%and(87.2±2.6)%.During the degradation,significant morphological changes were noticed.The plasmids could be released in a multiphasic fashion.Enhanced green fluorescent protein and Real-Time PCR analysis showed that genes were expressed in chondrocytes.lasting near 30 days.MTT indicated that the cotransinfection promoted the chondrocytes'proliferation. Conclusion IL-1Ra and TGF-β1 genes cotransfected into chondrocytes via chitosan-DNA microsphere could be expressed in a long term and cotransinfection promoted the chondroeytes'proliferation,which is the base of inhibiting the degeneration of cartilage and promote cartilage repair.
5.Enhancing the accumulation of beta-amyrin in Saccharomyces cerevisiae by co-expression of Glycyrrhiza uralensis squalene synthase 1 and beta-amyrin synthase genes.
Ying LIU ; Hong-Hao CHEN ; Hao WEN ; Ya GAO ; Li-Qiang WANG ; Chun-Sheng LIU
Acta Pharmaceutica Sinica 2014;49(5):734-741
Glycyrrhiza uralensis Fisch. ex DC is widely used in traditional Chinese medicine (TCM). Among its various active components, glycyrrhizic acid is believed to be the marker component. Squalene synthase (SQS) and beta-amyrin synthase (beta-AS) are key enzymes in the biosynthetic pathway of glycyrrhizic acid in G uralensis. To reveal the effects of co-expression of SQS1 and beta-AS genes on this pathway, 7 yeast expression vectors harboring different SQS1 variants and beta-AS were constructed and expressed in Saccharomyces cerevisiae as fusion proteins. TLC and GC-MS results showed that co-expression of SQS1 and beta-AS enhanced the accumulation of beta-amyrin. The effects of SQS12 were more obvious than the other two SQS1 variants. This study is significant for further investigations concerned with exploring the biosynthesis of glycyrrhizic acid in vitro and strengthening the efficacy of G. uralensis by means of increasing the content of glycyrrhizic acid.
Farnesyl-Diphosphate Farnesyltransferase
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genetics
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metabolism
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Glycyrrhiza uralensis
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genetics
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Intramolecular Transferases
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metabolism
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Oleanolic Acid
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analogs & derivatives
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metabolism
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Plant Proteins
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genetics
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Recombinant Proteins
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metabolism
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Saccharomyces cerevisiae
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metabolism
6.Comparison of homemade and imported HbsAg ELISA kits on screening blood samples.
Fu-ping LIU ; Jing-chun LIU ; De-wen WANG
Chinese Journal of Experimental and Clinical Virology 2006;20(2):84-86
BACKGROUNDTo evaluate homemade and imported HbsAg ELISA kits on screening blood donors.
METHODSSamples for evaluation included 120 HbsAg serum plates for the golden criteria and 400 sets of serum from blood donors in Dongguan. The samples underwent blind screening with homemade and imported ELISA kits respectively.
RESULTSThe sensitivity of homemade (Xinchuang) and imported (Diasorin) HbsAg ELISA kit were 85.71% (72/84) and 100% (84/84), respectively. Their specificity was 100% (436/436) and 96.55% (421/436) respectively. The consistency of two ELISA kits was 100%.
CONCLUSIONThe imported ELISA kit had the highest sensitivity, but its specificity was not as good as that of homemade ELISA kit. The two kinds of ELISA kits had good repetition. The combination of the two reagents may ensure the safety of blood transfusion.
Blood Donors ; China ; Enzyme-Linked Immunosorbent Assay ; instrumentation ; methods ; standards ; Hepatitis B ; blood ; diagnosis ; prevention & control ; Hepatitis B Surface Antigens ; blood ; Humans ; Mass Screening ; methods ; standards ; Reagent Kits, Diagnostic ; standards ; Reference Standards ; Reproducibility of Results ; Sensitivity and Specificity
7.Diagnosis and treatment of ovotesticular disorder of sex development: A report of 2 cases.
Jing-qi WANG ; Dong-wen WANG ; Chun LIU ; Hua YANG ; Ji-xiu XU
National Journal of Andrology 2015;21(10):917-920
OBJECTIVETo investigate the characteristics, diagnosis, and treatment of ovotesticular disorder of sex development (OT-DSD).
METHODSWe retrospectively analyzed 2 cases of OT-DSD treated in our hospital. The patients were 19 and 15 years old, respectively, and both received systematic physical examination and examinations of the karyotype, sex hormone, adrenocorticotropic hormone (ACTH), color Doppler ultrasonography, urethrocystoscopy, and human chorionic gonadotropin (HCG) test. Under the laparoscope, we performed surgical gonad exploration, gonadectomy, and vulvar orthopedics. Intraoperative exploration and pathology confirmed true hermaphroditism in both cases, with sex selection as female. One underwent laparoscopic resection of the ovotestis, and the other removal of the testis with the ovarian tissue reserved.
RESULTSThe patients were followed up for 12 months postoperatively, which found no abnormality in either the vulvas or the genital glands.
CONCLUSIONSurgical exploration of the gonad is the only method for the diagnosis of OT-DSD and sex selection is the key to treatment. Laparoscopic surgical exploration of the gonad and vulvar orthopedics are the first treatment options.
Adolescent ; Chorionic Gonadotropin ; Female ; Gonadal Steroid Hormones ; Humans ; Karyotype ; Laparoscopy ; Male ; Ovary ; Ovotesticular Disorders of Sex Development ; diagnosis ; surgery ; Retrospective Studies ; Testis ; surgery ; Young Adult
8.ITS sequences variation and phylogenetic analysis on 31 geographical populations of Notopterygium incisum.
Lu-cun YANG ; He-chun LIU ; Xue-li ZHOU ; Wen-hua XU ; Guo-ying ZHOU
China Journal of Chinese Materia Medica 2015;40(19):3748-3753
In this study, 31 Notopterygium incisum populations were analyzed using ITS sequences to investigate the genetic structure. The results showed that: the ITS region ranged in size from 634 to 635 bp and base composition was with high G + C content of 57.8%. Thirty-one polymorphic sites were detected from 402 sequences of 31 populations of N. incisum, and the proportion of polymorphic sites was 4.88%, in which parsimony informative sites were up to 12. And 31 haplotypes were identified based on these polymorphic sites. Molecular variance analysis (AMOVA) indicated that high genetic differentiation (57%) existed among population, and gene flow was low (N(m) = 0.38) among populations. Phylogenetic relationships of 31 haplotypes were analyzed using NJ method with N. forbesiias an out-group. Phylogenetic analysis showed that 31 haplotypes from different populations mixed together and did not form distinct geographically separated clades.
Apiaceae
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classification
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genetics
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Base Sequence
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China
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DNA, Intergenic
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genetics
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Gene Flow
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Genetic Variation
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Molecular Sequence Data
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Phylogeny
9.Effect of metoprolol on myocardial apoptosis after coronary microembolization in rats
Qiang SU ; Lang LI ; Yang-Chun LIU ; You ZHOU ; Wei-Ming WEN
World Journal of Emergency Medicine 2013;4(2):138-143
BACKGROUND: Coronary microembolization (CME) is a serious complication following percutaneous coronary intervention (PCI) in patients with acute coronary syndromes. The use of metoprolol before PCI can significantly protect ischemic myocardium from myocardial damage, but the function of metoprolol in the treatment of CME is not entirely clear. This study was to explore the effect and significance of metoprolol on myocardial apoptosis and caspase-3 activation after CME in rats. METHODS: Thirty rats were randomly divided into three groups including sham-operation (control group), CME plus saline (CME group), CME plus metoprolol (metoprolol group), 10 rats for each group. The CME group was induced by injecting 3000 polyethylene microspheres (42 μm) into the left ventricle during a 10-second occlusion of the ascending aorta; the control group was injected with physiological saline instead of microembolization ball; the metoprolol or saline group was given three intravenous bolus injections before CME. Echocardiography, TUNEL staining, and Western blotting were used to evaluate cardiac function, proportion of apoptotic cells and activation of caspase-3 respectively at 6 hours after operation. RESULTS: Echocardiographic parameters displayed that the metoprolol group improved cardiac function significantly compared with the CME group (P<0.05). The myocardial apoptotic rate of the CME group as wel as the contents of activated caspase-3 increased significantly (P<0.05), both of which were ameliorated significantly by metoprolol treatment (P<0.05). CONCLUSIONS: This study demonstrates that metoprolol can protect the myocardium during CME in rats by inhibiting apoptosis and improving cardiac function. These results suggest that the inhibition of apoptosis can be a potential therapeutic strategy for the treatment of CME.
10.Effect of team-based individualized health management on health literacy and health status among hypertensive patients
Xiuqin WEN ; Juhong LIU ; Jie ZHAO ; Chun CHANG ; Qingqi ZENG ; Yunting ZHENG
Chinese Journal of Health Management 2015;(4):285-290
Objective To explore the effect of team-based individualized health management on health literacy and health status among hypertensive patients in community. Method This study was a self-controlled quasi-experimental design. A two-stage cluster random sampling method was used. Two neighborhood committees among 23 neighborhood committees in Desheng community were randomly sampled. The name list of hypertensive patients were obtained from the 2 sampled neighborhood committees, getting one from each five, 470 in total before intervention and 444 in total after. The two groups of patients before and after intervention had comparability in the social demographic characteristics as the age (67.16± 9.84, 66.47 ± 10.50 years), the gender (the male accounted for 47.00%and 44.10%), the ethnicity (the Han accounted for 93.80%and 92.30%) and the marital status(the married accounted for 91.30%, 88.90%)(t=1.447 χ2 =0.774, 1.595, 2.555, P>0.05). Since July 2013, Desheng community health service center intervened the hypertensive patients by team-based individualized health management. After one year, we compared the change of their health literacy and health status. Result After health management, the proportion of hypertensive patients with adequate health literacy increased from 38.50% to 53.40%, The average rate of health knowledge awareness among them increased from 65.30% to 75.30%. The average accuracy of health services utilization skills of them increased from 49.10%to 70.90%, lifestyle behaviors in dietary and in exercise the average increase was from 35.55, 17.07 to 37.05, 21.28 respectively, after the intervention the patients did better work in self management behavior, such as self-test of blood pressure, medication compliance and control of salt intake (74.40%, 87.60%, 80.00%) were obviously better than before intervention (61.30%, 68.30%, 62.20%), and the differences were statistically significant (χ2 =16.813, 48.295, 52.058, P<0.05). After health management, the self-report health was better among hypertensive patients. The proportion of them with normalized blood pressure increased from 72.90% to 79.50%, and the difference was statistically significant (P<0.05). Conclusion Team-based individualized health management could increase health literacy of hypertensive patients in community and improve their health status obviously.