1.Study on oriented differentiation of bone marrow mesenchymal stem cells by fibroblast in rat uterine ligament with mechanical stretch
Chenchen REN ; Ruifang REN ; Bing ZHAO ; Xi ZHANG ; Yongjie JIANG
Chinese Journal of Obstetrics and Gynecology 2011;46(7):527-532
Objective To investigate the effect on the differentiation of bone marrow mesenchymal stem cells (BMSC) with non-contact co-culture with mechanical stimulated ligament fibroblasts. Methods A cyclic 10% uniaxia strain at 1 Hz was applied on rat pelvic ligament fibroblasts, then were co-cultured with BMSC for 3, 6 and 12 days in non-contact condition. The protein expression of collagen Ⅰ ,Ⅲ in BMSC were detected by SP method and revealed by the mean gray value. The mRNA expressions of collagens type Ⅰ and type Ⅲ in the BMSCs were measured with real-time (RT)-PCR ,and the results were indicated by the ratio between the mRNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) . Results (1) Protein expression; after 3 days co-culture with pelvic ligament fibroblasts, expression of collagen Ⅰ and Ⅲ in BMSC are 82. 4 ± 3. 4 and 76. 8 ± 2. 5. When compared with 80. 2 ± 2. 6 and 74. 6 ± 1. 1 in BMSC without co-culture, there was no significant difference (P > 0. 05) . After 6 days co-culture with pelvic ligament fibroblasts, the expression of collagen Ⅰ and Ⅲ of 126. 6 ±2. 2 and 118. 6 ± 1. 4 in BMSC were significantly higher than 82. 7 ±3. 0 and 76. 2 ± 1. 3 in BMSC without co-culture (P < 0. 05). Similarly, after 12 days co-culture with pelvic ligament fibroblasts, the expression of collagen Ⅰ and Ⅲ of 135. 3 ±3. 4 and 128. 7 ± 2. 6 in BMSC were significantly higher than 86. 6 ± 1. 3 and 81. 8 ± 1.4 in BMSC without co-culture (P <0.05). (2)mRNA expression:after 3 days co-culture with pelvic ligament fibroblasts , the mRNA expression of type Ⅰ and type Ⅲ collagens in BMSC are 2. 10 ±0. 20 and 1. 20 ±0. 30. When compared with mRNA expression of 2. 01 ±0. 12 and 1. 13 ±0.21 in BMSC without co-culture, no significant difference were observed (P > 0. 05). After 6 days co-culture with pelvic ligament fibroblasts , the mRNA expressions of type Ⅰ and type Ⅲ collagens mRNA were 5. 60 ±0. 21 and 2. 61 ±0. 20, which were significantly higher than 3. 70 ±0. 33 and 1. 82 ± 0. 14 in BMSC without coculture (P < 0. 05). After 12 days co-culture with pelvic ligament fibroblasts, the mRNA expressions of type Ⅰ and type Ⅲ collagens of 5. 91 ±0.31 and 2. 92 ±0. 23 were significantly higher than 4. 04 ±0. 21 and 2. 04 ±0. 13 in BMSC without co-culture (P <0. 05). Conclusion Non-contact co-culture with mechanical stretch stimulated ligament fibroblasts, it might promote synthesis of types Ⅰ and Ⅲ collagen in rat BMSCs and induced BMSC differentiated into pelvic ligament fibroblasts.
2.Current situation and requirements of quality control of blood disease laboratory in Chongqing:a cross sectional study
Jingkang XIONG ; Xi ZHANG ; Cheng ZHANG ; Xiangui PENG ; Jiashun REN
Chongqing Medicine 2016;45(30):4271-4273
Objective To understand the current situation and requirements of laboratory quality control of blood diseases in Chongqing .Methods A self‐designed questionnaire was designed ,and the investigation was carried out in 60 hospitals of Chongqing district .The main contents of the investigation were related to the instruments of work environment ,the regulations ,the conditions of the personnel ,etc .Results A total of 60 questionnaires were distributed ,57 questionnaires were valid .In the study ,only 14 .04%laboratories were the blood disease specialist laboratory ,most hospitals could complete the basic inspection items .66 .67% hospitals had all the experimental procedures and SOP documents .Most of staffs were with intermediate professional titles and undergraduate education .A total of 35 .08% experimental equipments with the value of over 5 million RMB ,45 .61% equipments had a special in‐spection records .About 45 .61% test reports would refer to the clinical performance ,and had a strict signing guideline .Most of the research objects reflected the problems ,including the backward equipments and the lack of personnel ,and hoped to set up telemedi‐cine indemnity system and carry out further education .Conclusion The overall quality of the laboratory for blood diseases in Chongqing is better ,it is recommended to strengthen the full quality control ,to establish emergency system ,and to improve the quality and level of laboratory .
4.Improved RP-HPLC method for determination of ferulic acid concentration in human serum
Jingqun HUANG ; Xi HUANG ; Li ZHANG ; Rui XU ; Ping REN
Chinese Traditional and Herbal Drugs 1994;0(09):-
Object To develop an improved RP-HPLC method for determining ferulic acid in the human serum. Methods The determination was carried on RP-HPLC, using Kromasil-C 18column, methanol-water-acetic acid (36.4∶63∶0.6) as mobile phase with a flow rate of 1.0 mL/min and at the detection wavelength of 322 nm, and p-hydroxybenzaldehyde was used as internal standard. The serum sample was deproteinized with acetonitrile to extract ferulic acid. Results The calibration curve showed good linearity over the range of 9.94-159.04 ng/mL (r=0.992 5). RSD was less than 10% within day and day-to-day, the average recovery was 99.77% and the minimal concentration in serum was 5 ng/mL. Conclusion This method, which is simple, rapid, sensitive, reproducible and low toxic, is appropriate for the quantitative determination of ferulic acid in the human serum.
5.Association of core binding factor alpha 1 and estrogen receptor alpha gene polymorphism with bone development
Yue ZHANG ; Lijun WANG ; Fu REN ; Huanjiu XI ; Yingnan FAN
Chinese Journal of Tissue Engineering Research 2009;13(46):9159-9162
Produced by bone cells and stored in the matrix.bone ceil growth factor can contribute to the regulation of bone growth.Gene regulation has its role in bone development.Many factors of this kind have been founded recently.They play their role during the bone formation and absorption processes in the way of autocrine and paracrine.The genes of core binding factor α_1(Cbfα_1)and estrogen receptor α(Erα)have been the recent years'hot factors that are related to the bone development,Gene polymorphism refers to variation on gene level which often occurs in the non-coding domain or the domain bearing no important regulatory function in gene order.This article reviewed the research status of the relation between Cbfα_1 and Erα gene polymorphism with bone development.
6.Effects of Deep Electroacupuncture on Cartilage in Knee Osteoarthritis Rabbits
Nini FU ; Xuezhi LI ; Fei LIU ; Xiaofang XI ; Yi REN ; Xiaoguang YANG ; Yu ZHANG
Chinese Journal of Rehabilitation Theory and Practice 2016;22(1):38-45
Objective To observe the effects of deep electroacupuncture on carlilage tissue in knee osteoarthritis (KOA) rabbits. Meth-ods 40 New Zealand rabbits were randomly divided into normal group (A, n=10) and model group (n=30). The model group was modeled KOA with Hulth-Telhag way, and identified with X-ray. Then they were divided into no-treated group (B, n=10), deep electroacupuncture group (C, n=10) and routine electroacupuncture group (D, n=10) randomly. The groups C and D accepted electroacupuncture since 6 weeks after modeling, for 4 weeks. They were measured with pH of joint fluid, observed structure and pathology of cartilage under transmission electron microscope, detected apoptosis index, and determined the expression of acid-sensing ion channel 1 (ASIC1), p38 mitogen-activated protein kinases (p38MAPK) and p53 with Western blotting, and distribution of ASIC1 with immunohistochemistry in cartilage tissue. Re-sults The pHs of joint fluid from high to low were ranged as the groups A=C>D>B (P<0.01). The cartilage structure was more complete in the groups A, C and D than in the group B. The apoptosis rates from less to more were ranged as the groups A=C
7.Ursolic acid down-regulates COX-2 expression by suppressing the activation of ERK in A549 cells
Jingsong WANG ; Jing SHEN ; Ting ZHANG ; Cong TANG ; Tiannian REN ; Tao XI
Journal of China Pharmaceutical University 2011;42(1):68-72
The antitumor effect and the mechanism of action of ursolic acid in A549 cells(human non-small-cell lung cancer cells) was investigated in this paper. Firstly, MTT assay was performed to test whether ursolic acid could inhibit the growth of A549 cells. Secondly, Western blot was utilized to measure the expression level of COX-2 and the activation of MAPKs. The MTI assay revealed that ursolic acid inhibited the growth of A549 cells. The result of Western blot suggested that ursolic acid inhibited the expression of COX-2 and activation of ERK and ERK specific inhibitor PD98059 suppressed the expression of COX-2 synergistically with ursolic acid. Taken together, our data suggest that ursolic acid can suppress LPS-induced COX-2 expression in A549 cells, which could be due to the inhibition of the activation of ERK.
8.The relationship between marOR mutations and the antibiotic resistance in Shigella spp
Jingchao REN ; Guangcai DUAN ; Haiyan YANG ; Ruili Lü ; Weidong ZHANG ; Yuanlin. XI
Chinese Journal of Microbiology and Immunology 2010;30(11):1044-1047
Objective To detect the influence of marOR mutations on antibiotic resistance in Shigella spp. Methods The marOR gene with four-base deletion was amplified by overlap PCR, then inserted in a T-vector and transformed into DH5α. The clone of marOR gene with four-base deletion and three point mutations was prepared from the strain having these mutations. Electrophoresis and sequencing were preformed to certify the correction of the cloned genes. Drug susceptibility tests were preformed for the strains harbouring the different clones [DH5α, DH5α (T), DH5α (marOR), DH5α (marOR-CATT), DH5α(marOR-CATT + 3m)]. Results Compared with the control strain (DH5α-T), the antibiotic resistances of marOR with four-base deletion [DH5α (marOR-CATT)] were higher to streptomycin, tobramycin, cefazolin and cefalexin, and the antibiotic resistances of marOR with four-base deletion and three point mutations [DH5α (marOR-CATT + 3m)] were higher to streptomycin and to tetracycline. The antibiotic resistances of DH5α (marOR-CATT) and DH5α (marOR-CATT +3m) to streptomycin, tetracycline, chloramphenicol, cefazolin, levofloxacin, ciprofloxacin and norfloxacin were higher than DH5α (marOR). The diameters of the antibiotics except the trimethoprim between DH5α (marOR-CATT) and DH5α (marORCATT +3m) had not significant disparity. Conclusion The four-base deletion in 1376-1379 sites of the marOR gene increased the resistance of Shigella spp to some antibiotics. The point mutations in 1411, 1417,1435 sites of the marOR gene have little influence on the antibiotic resistance of Shigella spp.
9.Detection of sex hormone and antisperm antibody in cryptorchidism boys after orchiopexy
Cuiping SONG ; Guolin ZHANG ; Jinlan CHEN ; Yanfang REN ; Kaisheng LV ; Zhiqiang ZHAO ; Zhan XI
Chinese Journal of Postgraduates of Medicine 2006;0(14):-
Objective To research the possible influence of operation and human chorionic gonadotropin(HCG) therapy on the development of antisperm antibody (AsAb) and serum levels of estradiol (E_2) and testosterone (T) of cryptorchidism boys after orchiopexy. Methods Forty cryptorchidism boys were studied who were divided into HCG treatment group and without HCG treatment group. Twenty boys who underwent inguinal operation were studied as the operation control group and 20 normal boys as the control group. The serum were collected for detection AsAb, E_2,and T. Results There were no significant difference in serum levels of E_2 and T among the three groups(P 0.05 ). Conclusions The level of AsAb is associated with the destroy of blood barrier and the disorder of immune modulation and endocrine. Treatment with HCG has no effect on AsAb level.
10.THE PROMOTION OF NEUROGENESIS IN SUBVENTRICULAR ZONE BY BONE MARROW STROMAL CELLS AFTER PERMANENT FOCAL CEREBRAL ISCHEMIA IN RATS
Pushuai WEN ; Huanjiu XI ; Jing GAO ; Pengfei ZHANG ; Fu REN ; Youfeng WEN
Acta Anatomica Sinica 2002;0(05):-
Objective To investigate the effect of transplanted bone marrow stromal cells on the cells in the subventricular zone of the rats with permanent focal cerebral ischemia and analyze cell types.Methods Permanent focal cerebral ischemia models were established with middle cerebral artery occlusion(MCAO) and divided into three groups: MCAO alone,intravenous infusion of 1ml PBS at 24 hours after MCAO,and intravenous infusion of 2?106 BMSCs 24 hours after MCAO.Then,the groups were subdivided into 7-day and 14-day groups after MACO.Neurological functions were detected by Zausinger evaluation;meanwhile,5-bromodeoxyuridine was injected to label the proliferating cells in the subventricular zone,and double-immunofluscent technologies were used to identify the cell type.Results On the 7th day and the 14th day after MACO,neurological functional scores of BMSCs-treated group were higher than those of the other two groups(P