1.Age-associated proliferation and differentiation changes of rat bone marrow mesenchymal stem cells
Hairui LI ; Dong ZHENG ; Can JIANG ; Jun GUO ; Aidong ZHANG ; Zicheng LI
Chinese Journal of Tissue Engineering Research 2015;(1):18-23
BACKGROUND:Autologous bone marrow mesenchymal stem cel transplantation for the treatment of cardiovascular disease has become one of the hotspots, but it is unclear whether the proliferation and directional differentiation of autologous bone marrow mesenchymal stem cels varies changes with age. OBJECTIVE: To explore the proliferation and differentiation changes of rat bone mesenchymal stem cels in different ages. METHODS:The bone marrow mesenchymal stem cels from Sprague-Dawley rats in different age groups were purified and cultured, and then examined by flow cytometry in terms of cel cycle. Meanwhile, neonatal rat cardiomyocytes were co-cultured with bone marrow mesenchymal stem cels. The morphologic changes of bone marrow mesenchymal stem cels and the protein expression of troponin T were detected with immunofluorescence technique. RESULTS AND CONCLUSION: The percentage of bone marrow mesenchymal stem cels in G0/G1 phase was increased with age; while the percentage of expression of troponin T proteins-positive bone marrow mesenchymal stem cels were decreased with age. These findings indicate that the proliferation and differentiation abilities of rat bone marrow mesenchymal stem cels descend with age.
2.Intestinal Function Changes in Mice after Spinal Cord Injury in Early Stage
Yu LIANG ; Can LI ; Lan LUO ; Zheng YANG ; Qiang WANG ; Jun MA ; Yangyang WANG ; Lulu ZHANG ; Wei YANG ; Fang YI
Chinese Journal of Rehabilitation Theory and Practice 2015;21(2):148-152
Objective To investigate the influence of spinal cord injury on intestinal function in mice after a short period. Methods 105 Kunming mice were randomly assigned to normal group (group A, n=30), sham group (group B, n=30) and model group (group C, n=45). Group A received no treatment, group B was only exposed the spinal cord, while group C was induced by a constant compression with a modified arterial forcep at T10 to establish spinal cord injury model. 12 hours, 24 hours and 48 hours after modeling, myoelectric slow wave activity and tension activity of the ileum were tested, and HE staining was used. Results Compared with groups A and B, the slow wave activity was significantly weaker in group C at every time point (P<0.05), so was the amplitude of tension activity (P<0.05). Frequency of tension activity was obviously higher in group C than in groups A and B 24 hours and 48 hours after modeling (P<0.05). The injury scores were higher in group C than in groups A and B (P<0.05). Conclusion There is reduce in myoelectric slow wave activity, tension activity of the ileum and mild injury in intestinal mucosa in mice after spinal cord injury in the early stage.
3.MRI diagnosis of benign sacrococcygeal teratomas of the infants.
Can LAI ; Shi-zheng ZHANG ; Hua-jun YANG
Journal of Zhejiang University. Medical sciences 2006;35(1):103-107
OBJECTIVETo investigate the value of MR imaging in the diagnosis of benign sacrococcygeal teratomas of the infants.
METHODSMR imaging of benign sacrococcygeal teratomas in 6 cases proved by surgery and pathology was retrospectively reviewed. In all patients, a fast imaging sequence, fast spin echo sequence was employed, together with short time inversion recovery sequence and contrast enhancement scanning by intravenous injection of Gd-DTPA.
RESULTSThere were 6 benign sacrococcygeal teratomas, which were heterogeneous masses and arose from the distal sacrococcygeal region in the pelvis. The MR imaging appearances of the benign sacrococcygeal teratomas were characteristic, T1- and T2-weighted images demonstrated a large mass containing round, well-defined areas of varying signal intensity representing its cystic, solid, and sometimes fat, calcification within the lesions.
CONCLUSIONMR imaging provides definitive information of benign sacrococcygeal teratomas and clearly shows both extra-and intra-pelvic components, and even better anatomic details, which facilitates the surgical planning adequately.
Female ; Humans ; Infant ; Infant, Newborn ; Magnetic Resonance Imaging ; Male ; Retroperitoneal Neoplasms ; diagnosis ; Sacrococcygeal Region ; Teratoma ; diagnosis
4.Efficacy of natural vitamin E on oligospermia and asthenospermia: a prospective multi-centered randomized controlled study of 106 cases.
Xiang-Feng CHEN ; Zheng LI ; Ping PING ; Ji-Can DAI ; Feng-Bin ZHANG ; Xue-Jun SHANG
National Journal of Andrology 2012;18(5):428-431
OBJECTIVETo explore the therapeutic effect of natural vitamin E (VitE) on oligospermia and asthenospermia in in- fertile men.
METHODSWe conducted a prospective multi-centered randomized controlled study on 64 infertile men with oligospermia (31 as controls treated with Tamoxifen 10 mg bid and 33 as experimental cases treated with Tamoxifen 10 mg bid + VitE 100 mg tid) and 42 cases of asthenospermia (20 as controls treated with Levocarnitine oral solution 1 bottle bid and 22 as experimental cases treated with Levocarnitine oral solution 1 bottle bid + VitE 100 mg tid). We compared the control and experimental groups in sperm concentration and percentage of progressively motile sperm before and 3 months after medication, as well as the rate of clinical pregnancy and adverse events.
RESULTSAmong the oligospermia patients, the average sperm concentrations in the control and experimental groups were 8.00 x 10(6)/ml and 10.66 x 10(6)/ml before medication (P > 0.05). After medication, the numbers of cases evaluated as with no, slight, moderate and marked improvement in sperm concentration were 10 and 9 (P > 0.05), 16 and 14 (P > 0.05), 5 and 4 (P > 0.05) and 0 and 0 (P >0.05); and the numbers of natural pregnancies were 0 and 6 in the control and experimental groups (P < 0.01). Among the asthenospermia patients, the average rates of progressively motile sperm were 17.00% and 18.10% in the control and experimental groups before medication (P > 0.05). After medication, the numbers of cases evaluated as with no, slight, moderate and marked improvement in the percentage of progressively motile sperm were 7 and 2 (P < 0.01), 4 and 8 (P < 0.01), 3 and 2 (P > 0.05) and 1 and 1 (P > 0.05), and the numbers of natural pregnancies were 5 and 9 in the two groups (P < 0.01), but no adverse events were observed.
CONCLUSIONAs a safe and effective adjuvant agent for the treatment of oligospermia and asthenospermia, vitamin E can improve sperm concentration, the percentage of progressively motile sperm, and finally the rate of natural pregnancy.
Adult ; Asthenozoospermia ; drug therapy ; Carnitine ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Oligospermia ; drug therapy ; Pregnancy ; Pregnancy Rate ; Prospective Studies ; Tamoxifen ; therapeutic use ; Treatment Outcome ; Vitamin E ; therapeutic use ; Young Adult
5.Artesunate suppresses human endometrial carcinoma RL95-2 cell proliferation by inducing cell apoptosis.
Jun-sheng ZHENG ; Ming-hua WANG ; Min HUANG ; Yun-peng LUO ; Can MI
Journal of Southern Medical University 2008;28(12):2221-2223
OBJECTIVETo investigate the inhibitory effect of artesunate on human endometrial carcinoma RL95-2 cell line proliferation in vitro and the possible mechanisms.
METHODSThe inhibitory effect of artesunate on the cell proliferation was assessed with MTT assay. Transmission electron miscrosopy was used to observe the morphological change of the cells after the treatment. Flow cytometry was performed to examine the changes in the cell cycle, reactive oxygen species (ROS) levels (with DCFH-DA labeling) and mitochondrial membrane potential (rhodamine123 staining), and caspase-3 activity was detected by immunohistochemistry.
RESULTSArtesunate inhibited the proliferation of RL95-2 cells with an IC(50) of 26.29 microg/ml. Transmission electron microscopy revealed early apoptotic changes of the cells with obvious chromatin fragmentation. The cell cycle arrest at G(0)/G(1) phase was observed by flow cytometry, and immunohistochemistry demonstrated caspase-3 positivity in cytoplasm. ROS generation in the cells increased obviously after treatment with artesunate for 72 h, which also resulted in lowered mitochondrial membrane potential.
CONCLUSIONArtesunate suppressed the proliferation of RL95-2 cells in vitro possibly by inducing cell apoptosis.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Artemisinins ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Endometrial Neoplasms ; pathology ; Female ; Humans
6.Development of time resolved fluorescence microparticle-conjugated anti-human procalcitonin antibody for homogeneous immunoassay
jun Can WU ; yu Qing L(U) ; jie Huai HAO ; na Cheng ZHAO ; ling Yu ZHENG ; qiang Yong JIANG
Military Medical Sciences 2017;41(8):689-693
Objective To develop europium (Ⅲ) [Eu (Ⅲ)] chelated microparticles for homogeneous immunoassay.Methods Anti-human PCT antibodies were labeled with Eu (Ⅲ) chelated nanoscale microparticles as the detection antibody,and another anti-human PCT antibody was labeled with biotin as the solid-phase antibody.Magnetic microspheres labeled with streptavidin were used to separate the complexes of Eu-IgM-PCT-IgM-Biotin.Results In the homogeneous immunoassay,the standard curve fit was not linear.The quadratic curve was Y =19170.12 + 75493.74X-26.00X2(r =0.9986).According to the standard curve,the limit of detection for PCT was 0.04 ng/ml.Conclusion The homogeneous immunoassay which uses Eu (Ⅲ) chelated microparticles is highly sensitive for detection of PCT recombinant antigens and may serve as a promising method to measure serum PCT levels in the future.
7.Bone marrow stromal cells transfected with ciliary neurotrophic factor gene ameliorates the symptoms and inflammation in C57BL/6 mice with experimental allergic encephalomyelitis.
Zheng-qi LU ; Xue-qiang HU ; Can-sheng ZHU ; Xue-ping ZHENG ; Dun-jing WAN ; Ran-yi LIU ; Bi-jun HUANG ; Wen-lin HUANG
Journal of Southern Medical University 2009;29(12):2355-2361
OBJECTIVETo investigate the anti-inflammatory effect of bone marrow stromal cells (MSCs) transfected with recombinant adenovirus-mediated ciliary neurotrophic factor (CNTF) gene in C57BL/6 mice with experimental allergic encephalomyelitis (EAE).
METHODSAn adenovirus vector containing CNTF gene Ad-CNTF-IRES-GFP was constructed and transfected in the MSCs (MSC-CNTF). After examination of CNTF expression, the transfected cells were transplanted in C57BL/6 mice with MOG 35-55-induced EAE, which were monitored for the changes in the symptoms scores. The levels of tumor necrosis factor-alpha (TNF-alpha), inteferon-gamma (IFN-gamma), interleukin-12P35 (IL-12P35), and IL-10 in the peripheral blood of the mice were detected, and the number of MSC-CNTF cells in the spleen and spinal cord was counted. CD3+ T cell infiltration and TNF-alpha and IFN-gamma expressions in the lesions were also observed after the cell transplantation.
RESULTSCNTF gene transfection resulted in significantly increased CNTF expression in the MSCs. The mice receiving MSC-CNTF transplantation exhibited significantly improved symptoms with shortened disease course and lessened disease severity. The cell transplantation also resulted in significantly decreased peripheral blood TNF-alpha levels, ameliorated CD3+T cell infiltrations and lowered TNF-alpha expression in the lesions, while the levels of IFN-gamma underwent no significant changes.
CONCLUSIONTransplantation of CNTF gene-transfected MSCs results in decreased peripheral blood TNF-alpha and IFN-gamma levels and reduced inflammatory cells, CD3-positive cells and TNF-alpha expression in the lesion of EAE, therefore providing better effect than MSCs in relieving the symptoms of EAE in mice.
Adenoviridae ; genetics ; metabolism ; Animals ; Bone Marrow Cells ; metabolism ; Ciliary Neurotrophic Factor ; biosynthesis ; genetics ; therapeutic use ; Encephalomyelitis, Autoimmune, Experimental ; therapy ; Female ; Genetic Therapy ; Interferon-gamma ; blood ; Mice ; Mice, Inbred C57BL ; Random Allocation ; Stromal Cells ; metabolism ; T-Lymphocytes ; immunology ; Transfection ; Tumor Necrosis Factor-alpha ; blood
8.Clinical significance of MP-DNA from endotracheal aspirates in diagnosis of Mycoplasma pneumoniae pneumonia in children.
Wen-Jing GU ; Xin-Xing ZHANG ; Zheng-Rong CHEN ; Yong-Dong YAN ; Can-Hong ZHU ; Li HUANG ; Mei-Juan WANG ; Xue-Jun SHAO ; Wei JI
Chinese Journal of Contemporary Pediatrics 2015;17(9):937-941
OBJECTIVETo compare the detection rates of Mycoplasma pneumoniae (MP) from nasopharyngeal aspirates (NPA) and bronchoalveolar lavage fluid (BALF) in children with pneumonia.
METHODSA total of 164 hospitalized children with pneumonia were enrolled. NPA and BALF of these children were collected within 24 hours of admission, and MP-DNA was detected by fluorescence quantitative PCR. Venous blood samples of all these children were collected within 24 hours of admission and on days 7-10 of treatment, and serum MP-IgM was detected using ELISA.
RESULTSThe positive rate of MP-DNA in NAP of the 164 cases was 51.8% , which was lower than 63.4% as the detection rate of MP-IgM in serum (P=0.044), and the two detection rates were moderately consistent with each other (Kappa=0.618, P<0.01). The positive rate of MP in BALF was 71.3%, which was not significantly different with that of MP-IgM in serum (P>0.05), and the detection rates were well consistent (Kappa=0.793, P<0.01). The detection rate of MP in NPA was lower than that in BALF (P<0.01), with moderate consistency between two of them (Kappa=0.529, P<0.01). The median MP copy number in BALF was significantly higher than that in NPA (P<0.01). The MP detection rates in NPA and BALF were significantly different among different courses of disease (P<0.05). As the course of disease extended, the MP detection rates in both NPA and BALF showed a declining trend; children with MP pneumonia of 1-2 weeks' duration and 2-4 weeks' duration had a higher MP-DNA detection rate in BALF than in NPA (P<0.05).
CONCLUSIONSMP-DNA in BALF has a high sensitivity, with a great significance for early diagnosis of MP pneumonia, while NPA MP-DNA tests may lead to a missed diagnosis.
Adolescent ; Bronchoalveolar Lavage Fluid ; microbiology ; Child ; Child, Preschool ; DNA, Bacterial ; analysis ; Female ; Humans ; Infant ; Male ; Nasopharynx ; microbiology ; Pneumonia, Mycoplasma ; diagnosis
9.A study on relationship between variation of P gene region and genotyping of Lamivudine-resistant hepatitis B virus
Guo-Can YANG ; Guo-Feng MAO ; Shao-Qing SHEN ; Zhuan ZHENG ; Jian-Jun CHEN
Journal of Preventive Medicine 2014;(6):544-547
Objective To analyze hepatitis B virus( HBV)genotype and the correlation between variation of P gene region and genotyping of Lamivudine-resistant HBV,in order to provide molecular virology basis for anti-virus individualized treatment to different HBV genotypes. Methods Serum markers and HBV DNA of 187 Lamivudine -resistant chronic HBV patients were detected by ELISA and fluorescent quantitative polymerase chain reaction. Moreover,DNA Star and viral genotyping tool were used. Results Of the 187 cases infected with HBV genotype B and C,51 cases were B gene (27. 27%),134 cases were C gene(71. 66%),and 2 cases were mixed infections(1. 07%). HBeAg levels were significantly different between genotype B and C(P<0. 05). P gene mutation results showed that genotype B dominated by YMDD mutation alone and genotype C dominated by YMDD and rtL180M mutation. Conclusion The main HBV genotype of this study is genotype C,and different genotypes may determine the variation patterns of P region which associated with resistance.
10.Preparation of Fe3O4@SW-MIL-101-NH2 for selective pre-concentration of chlorogenic acid metabolites in rat plasma,urine,and feces samples
Yin SHI-JUN ; Zhou XI ; Peng LI-JING ; Li FANG ; Zheng GUO-CAN ; Yang FENG-QING ; Hu YUAN-JIA
Journal of Pharmaceutical Analysis 2022;12(4):617-626
An innovative sandwich-structural Fe-based metal-organic framework magnetic material(Fe3O4@SW-MIL-101-NH2)was fabricated using a facile solvothermal method.The characteristic properties of the material were investigated by field emission scanning electron microscopy,transmission electron mi-croscopy(TEM),energy-dispersive X-ray spectroscopy,Fourier transform infrared spectroscopy,X-ray powder diffraction,vibrating sample magnetometry,and Brunauer-Emmett-Teller measurements.Fe3O4@SW-MIL-101-NH2 is associated with advantages,such as robust magnetic properties,high specific surface area,and satisfactory storage stability,as well as good selective recognition ability for chlorogenic acid(CA)and its metabolites via chelation,hydrogen bonding,and π-interaction.The results of the static adsorption experiment indicated that Fe3O4@SW-MIL-101-NH2 possessed a high adsorption capacity toward CA and its isomers,cryptochlorogenic acid(CCA)and neochlorogenic acid(NCA),and the adsorption behaviors were fitted using the Langmuir adsorption isotherm model.Then,a strategy using magnetic solid-phase extraction(MSPE)and ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry(UPLC-Q-TOF MS/MS)was developed and suc-cessfully employed for the selective pre-concentration and rapid identification of CA metabolites in rat plasma,urine,and feces samples.This work presents a prospective strategy for the synthesis of magnetic adsorbents and the high-efficiency pretreatment of CA metabolites.