1.Mechanism of in vitro differentiation of bone marrow stromal cells into neuron-like cells.
Qian, CHU ; Yaping, WANG ; Xinqiao, FU ; Suming, ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2004;24(3):259-61
In order to study whether marrow stromal cells (MSCs) can be induced into nerve-like cells in vitro, and the mechanism, the MSCs in Wistar rats were isolated and cultured, and then induced with DMSO and BHA in vitro. The expression of specific marking proteins in neurons, glia and neural stem cells were detected before preinduction, at 24 h of preinduction, at 6 h, 24 h, and 48 h of neuronal induction by using immunohistochemistry and Western blotting. The ultrastructural changes after the inducement were observed. The results showed that after the inducement, many MSCs turned into bipolar, multipolar and taper, and then intersected as network structure. At the same time, some MSCs had the typical neuron-like ultrastructure. Immunohistochemistry revealed that NeuN and Nestin expression was detectable after inducement, but there was no GFAP and CNP expression. Western blotting showed the expression of Nestin was strong at 6 h of neuronal induction, and decreased at 24 h, 48 h of the induction. NeuN was detectable at 6 h of neuronal induction, and increased at 24 h, 48 h of the induction. It was concluded MSCs were induced into neural stem cells, and then differentiated into neuron-like cells in vitro.
Bone Marrow Cells/*cytology
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*Cell Differentiation
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Cells, Cultured
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Glial Fibrillary Acidic Protein/metabolism
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Neurons/*cytology
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Rats, Wistar
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Stromal Cells/cytology
2.Mechanism of differentiation of mesenchymal stem cells into neuron-like cells in vitro
Qian CHU ; Ya-ping WANG ; Xin-qiao FU ; Suming ZHANG
Chinese Journal of Rehabilitation Theory and Practice 2004;10(1):13-14
ObjectiveTo study the mechanism of differentiation of mesenchymal stem cells(MSCs) into neuron-like cells in vitro.MethodsMSCs of Wistar rats were separated and cultured, and then induced with DMSO and BHA in vitro. The specific marking proteins of neurons, glia and neural stem cells were detected before preinduction, at 24h after preinduction, at 6h, 24h, and 48h after neuronal induction.ResultsAfter the inducement, many MSCs turned into bipolar,multipolar and taper,and then intersected as network structure. Nestin was strong positive at 6h after neuronal induction, and decreased at 24h, 48h after the induction. NeuN was present at 6 h after neuronal induction, and increased at 24h, 48h after the induction.ConclusionMSCs can be induced into neural stem cells(NSCs) at first, and then differentiate into neuron-like cells in vitro.
3.Development of EV71, CA16 and other enterovirus vrial real-time qualitative PCR diagnostic kit.
Li-Qin LI ; Jing ZHONG ; Lin-Fu ZHOU ; Fu-Chu QIAN ; Jia-Wei WANG ; Li-Cheng DAI
Chinese Journal of Experimental and Clinical Virology 2013;27(3):224-227
OBJECTIVEA novel multiplex real-time RT-PCR kit was developed to detect EV71, CoxA16 and other human enteroviruses simultaneously with an internal amplification control to avoids false negatives, which used for hand, foot and mouth disease in the clinical diagnosis and epidemiological surveillance.
METHODSDesign specific primers and probes of EV71, CA16, other intestinal virus and internal amplification control, improve the extraction method of virus nucleic acid. Optimization the detection system of real-time quantitative PCR. Research the products of the accuracy, stability, precision, amplification efficiency and detection of linear range.
RESULTSThe primers and probes had high spicificity. The Viral RNA extraction effect of this Kit is as same as that of QIAamp Viral RNA mini Kit (QIAGEN company), but less reagent cost. The optimal concentrations of primers and probes are 0.2 micromol/L for all the upstream and downstream primers, 0.06 micromol/L for probes of other human enteroviruse, 0.08 micromol/L for probes of EV71 and CA16 respectively. The kit has good stability, accuracy and precision. The amplification efficiencies of EV71, CoxA16 and other human enteroviruses are 106% ,101% and 105% and the detection of linear range is from 10(9) copies/microl-10(2) copies/microl.
CONCLUSIONThe novel multiplex real-time RT-PCR kit for detecting EV71, CoxA16 and other human enteroviruses simultaneously with an internal amplification control has good stability, accuracy, precision and amplification efficiencies. So it has great value in clinical application.
Enterovirus ; isolation & purification ; Humans ; Reagent Kits, Diagnostic ; Real-Time Polymerase Chain Reaction ; methods
4.Mechanism of in vitro differentiation of bone marrow stromal cells into neuron-like cells.
Qian CHU ; Yaping WANG ; Xinqiao FU ; Suming ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2004;24(3):259-261
In order to study whether marrow stromal cells (MSCs) can be induced into nerve-like cells in vitro, and the mechanism, the MSCs in Wistar rats were isolated and cultured, and then induced with DMSO and BHA in vitro. The expression of specific marking proteins in neurons, glia and neural stem cells were detected before preinduction, at 24 h of preinduction, at 6 h, 24 h, and 48 h of neuronal induction by using immunohistochemistry and Western blotting. The ultrastructural changes after the inducement were observed. The results showed that after the inducement, many MSCs turned into bipolar, multipolar and taper, and then intersected as network structure. At the same time, some MSCs had the typical neuron-like ultrastructure. Immunohistochemistry revealed that NeuN and Nestin expression was detectable after inducement, but there was no GFAP and CNP expression. Western blotting showed the expression of Nestin was strong at 6 h of neuronal induction, and decreased at 24 h, 48 h of the induction. NeuN was detectable at 6 h of neuronal induction, and increased at 24 h, 48 h of the induction. It was concluded MSCs were induced into neural stem cells, and then differentiated into neuron-like cells in vitro.
Animals
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Bone Marrow Cells
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cytology
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Cell Differentiation
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Cells, Cultured
;
Glial Fibrillary Acidic Protein
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metabolism
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Neurons
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cytology
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Rats
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Rats, Wistar
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Stromal Cells
;
cytology
5.Expression characteristics of nuclear factor kappa B in hepatocellular carcinoma tissues.
Wei-min ZHOU ; Jin-liang PING ; Fu-chu QIAN ; Guo-lei ZHANG
Chinese Journal of Hepatology 2009;17(11):843-846
OBJECTIVETo investigate the expression characteristics of nuclear factor kappa B (NF-kB) in hepatocellular carcinoma (HCC) tissues and its correlation with tumor necrosis factor alpha (TNF alpha) and clinical pathological features.
METHODSThirty liver specimens from HCC patients were collected by self-control method. The localization and expression of NF-kappaB in HCC and their surrounding tissues were detected by immunohistochemistry and enzyme linked immunosorbent assay (ELISA), respectively. And the levels of TNF alpha in these tissues were analyzed by ELISA.
RESULTSThe expressed NF-kappaB was localized in nucleus and cytoplasm in HCC, whereas only in cytoplasm in the surrounding tissues. The expression level and density of NF-kappaB in HCC tissues were obviously higher than those in the surrounding tissues (P < 0.01), which was positively correlated with increased TNF alpha in HCC tissues (r = 0.964, P < 0.01). No positive correlation was found between NF-kappaB expression and histological differentiation grade, number of tumor, size of tumor, and HBsAg positive (P > 0.05).
CONCLUSIONThe expression and localization of NF-kappaB in HCC tissues are obviously different from those in the surrounding normal liver tissues, and the level of nucleoprotein NF-kappaB in HCC tissues is correlated with expressed TNF alpha, suggesting that TNF alpha can activate NF-kB, the activated NF-kB then translocates to the nucleus and plays important role in the carcinogenesis of HCC.
Adult ; Aged ; Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Nucleus ; metabolism ; Female ; Humans ; Immunohistochemistry ; Liver ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; NF-kappa B ; metabolism ; Signal Transduction ; Tumor Necrosis Factor-alpha ; metabolism
6.Correlations between preS1-antigen, HBV-DNA and HBV serum markers in patients with chronic hepatitis B.
Hui ZHOU ; Chu-wen JIANG ; Jing-lin QIAN ; Shi-jian LI ; Jie-ling LIANG ; Xue-fu CHEN
Journal of Southern Medical University 2008;28(7):1184-1186
OBJECTIVETo study the correlations between preS1 antigen, HBV-DNA and hepatitis B virus (HBV) serum markers in patients with chronic hepatitis B.
METHODSThe HBV markers, preS1 antigen and HBV-DNA were determined using enzyme- linked immunosorbent assay and quantitative PCR in 1158 patients with chronic hepatitis B.
RESULTSIn these patients, the HBV-DNA positivity rate was 68.9%, significantly higher than preS1 antigen positivity (54.8%, chi2=53.24, P<0.005). The positivity rates of both HBV-DNA and PreS1-antigen were significantly higher in HBeAg-positive patients than in HBeAg-negative patients (P<0.005). The coincident rates of preS1-antigen and HBeAg with HBV-DNA were 56.9% and 63.3%, respectively. PreS1 antigen had higher sensitivity but lower specificity than HBeAg. The detection rates of preS1 antigen and HBeAg increased with the level of HBV-DNA, and preS1 antigen positivity was higher than that of HBeAg in patients with low HBV-DNA levels.
CONCLUSIONDetection of HBV serum markers along with preS1 antigen and HBV-DNA may help assess the status of viral replication and therapeutic efficacy in patients with chronic hepatitis B. PreS1 antigen may serve as an auxiliary indicator in HBeAg-negative cases or when HBV-DNA detection is impossible.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; DNA, Viral ; blood ; Female ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; immunology ; isolation & purification ; Hepatitis B, Chronic ; blood ; immunology ; virology ; Humans ; Male ; Middle Aged ; Protein Precursors ; blood ; Virus Replication ; genetics ; Young Adult
7. Endoscopic surgery for thalamic hemorrhage breaking into ventricles: Comparison of endoscopic surgery, minimally invasive hematoma puncture, and external ventricular drainage
Chu-Hua FU ; Ning WANG ; Hua-Yun CHEN ; Qian-Xue CHEN
Chinese Journal of Traumatology 2019;22(6):333-339