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Objective To explore the different diagnostic values of acid-fast staining, tuberculosis culture, tuberculosis DNA detection (TB-DNA), tuberculosis RNA constant temperature amplification technology (SAT-TB) in the detection of Mycobacterium tuberculosis in sputum specimens. Methods A total of 200 pulmonary tuberculosis patients and 80 non-tuberculosis patients who were hospitalized in Hebei Chest Hospital between September 2015 and September 2019 were selected for this study. Sputum samples were collected after admission, and the detection values of acid-fast staining, tuberculosis culture, TB-DNA, and SAT-TB in sputum samples were statistically analyzed. Results The differences in the sensitivity, accuracy, positive predictive value, and negative predictive values of the four diagnostic methods of acid-fast staining, tuberculosis culture, TB-DNA, and SAT-TB were statistically significant (P TB-DNA> tuberculosis culture> acid-fast staining. In terms of the positive predictive value of diagnosis, the values of SAT-TB, TB-DNA, and tuberculosis culture were higher than that of acid-fast staining. The Kappa values of the four methods and the gold standard were: Kappa (acid-fast staining) = 0.145, Kappa (tuberculosis culture) = 0.395, Kappa (TB-DNA) = 0.602, and Kappa (SAT-TB) = 0.770. Conclusion The four diagnostic methods of acid-fast staining, tuberculosis culture, TB-DNA, and SAT-TB all had certain detection value with their advantages and disadvantages. SAT-TB was a better detection method with high specificity, good sensitivity, and a short detection timer, which could quickly identify bacteria and distinguish live bacteria.
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Objective To investigate the expression of miR-133b in the brain of methamphetamine(MA) dependent rats and its regulatory effects on neuronal toxic injury.MethodsThrough continuous intraperitoneal injection to rats with MA(10 mg/kg),the conditioned place preference(CPP) rats model was established,and the expression of miR-133b in the cerebral cortex of model rats was detected by real-time fluorescence quantitative PCR(RT-PCR).PC12 cells were cultured in vitro and treated with MA(800 μmol/L),and then miR-133b expression in cultured neurons was detected.miR-133b mimics and inhibitor were transfected to PC12 cells respectively to observe the effect of miR-133b on the mitochondrial membrane potential(MMP) of cultured neurons.ResultsAfter continuous intraperitoneal injection with MA for 14 days,the residence time of rats in the box with medicine((620.20±44.80)s) was significantly longer compared with the control group((341.80±25.12)s,P<0.01),which showed that MA dependent rats model was successfully established.The RT-PCR detection results showed that the expression of miR-133b in the cerebral cortex of model rats(0.36±0.05) significantly decreased compared with the control group(0.99±0.08,P<0.01).In the in vitro model,most of the neuronal cell bodies became round and the neuorites were withdrawn after MA treatment.Compared with the control group(1.00±0.02),the RT-PCR detection results showed that the expression of miR-133b in MA group(0.74±0.05) decreased(P<0.05).The JC-1 detection results showed that the MMP of the MA group(109.85±7.03) decreased significantly contrast to the control group(36.49±3.89,P<0.01),the MMP of the miR-133b mimics group(58.97±6.56) increased significantly contrast to the mimics control group(135.46±15.04,P<0.01) and the MMP of the miR-133b inhibitor group(162.84±14.15) decreased contrast to the inhibitor control group(139.81±12.26,P<0.05).ConclusionsThe expression of miR-133b in the cerebral cortex of MA dependent rats and in vitro neuron model treated with MA are significantly downregulated.By regulating the expression of miR-133b,the MMP damage of cultured neurons treated with MA is changed,indicating that miR-133b is not only involved in the nerve injury induced by MA,but also possiblely as a molecular target for intervention.
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[Objective]To investigate the expression change of miR-134 in methamphetamine(MA)-induced neuronal injury in PC12 cells and its effect on neuronal excitability and understand the pathogenesis of methamphetamine-induced neuronal injury.[Methods]PC12 cells in the logarithmic phase were divided into control group and MA group. The MA group was treated with 800μmol/L MA to establish the model of neuronal injury. The cellular injury was observed under microscope. The neuronal apoptosis was detected by Hoechst3342/PI double staining,and miR-134 expression was measured by using real-time quantitative PCR (Real time-PCR). Furthermore,we constructed miR-134 interference vector and observed its effect on evoked action potential.[Results]The cultured PC12 cells were damaged under the 800 μmol/LMA treatment ,and neurites became shorter ,the apoptotic cells were evidence. Real time-PCR showed that miR-134 expression was increased after MA treatment. Electrophysiological data showed that the evoked action potential increased after miR-134 interference.[Conclusions]High concentration of MA can induce neuronal damage and apoptosis and also increase miR-134 expression. While silence miR-134 expression can increase neuronal excitability.Our study provides an experimental basis for elucidating the possible mechanism of MA-induced neuronal injury and the role of miR-134 in neurotoxicity and neuronal excitability.
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Objective The aim of the study was to observe the survival and differentiation of embryonic neuroepithelial cells in lateral ventricle of rats after transplantation. Methods Embryonic rats(E12d) were obtained from the pregnant Sprague\|Dawley rats;neuroepithelial cells were dissociated from embryonic neural tube and treated with 0^25% trypsin,then transplanted into the lateral ventricle of father rat.The hosts were anesthetized and transcardially perfused with PBS followed by 4% paraformaldehyde fixative by 10?d,14?d,21?d and 28?d transplantation respectively.Coronal sections of the brain were cut with in cryostat microtome.Using the techniques of histology staining,Nissl staining and immunocytochemistry, survival of neuroepithelial cells transplant and neuro\|specific enolase (NSE) and glial fibrillary acidic protein (GFAP) expression of some stem cells were examined. Results Some grafts were found survival which attached to the ventricle by day 10.Some grafts could migrate into the third ventricle and grew into a mass of cells.Some stem cells were able to differentiate into pyramidal cells.By immunocytochemistry,a mass of NSE\|positive cells and GFAP\|positive cells could be detected in the core of graft region.Around intracerebral grafts,some astrocytes showed immunoreactive for glial fibrillary acidic protein.Conclusion\ These findings demonstrate that embryonic stem cells dissociated from neural tube can survive and differentiate into neurons and astrocytes.
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Objective To explore the survival,migration and differentiation of embryonic stem cells after transplantation into striatum and provide advantageous data for feasibility and safety of therapeutic transplantation. Methods We transplanted embryonic stem cells(undifferentiated ESCs and ESCs that had already developed into the stage of neural progenitor cells respectively) into striatum of the rat.Then differentiated cells were determined by morphological observation,Nissl's staining,TH and BrdU immunocytochemistry. Results We found that after transplanted 4 weeks,partially differentiated ESCs could survive and migrate into the surrounding host tissue.Some of them differentiated into TH-positive cells which had Nissl's bodies in cytoplasm.Whereas undifferentiated ESCs couldn't differentiate into TH-positive cells effectively and have the tendency to form tumor.Conclusion When conducting transplantation experiments of ESCs,it's better for ESCs to be induced into the stage of neural progenitor cells first and then transplanted.;
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Objective Embryonic neuroepithelial cells (NEC) were transplanted into the brains of Parkinson disease rats, the survival and differentiation of NEC were investigated. Methods Embryonic rats (E11) were obtained from the pregnant Wistar rats. Neuroepithelial cells were dissociated from embryonic neural tube and treated with 0
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Objective To isolate neuroepithelial stem cells from the spinal cord neural tube of the embryonic rat and induce them to differentiate into dopaminergic neurons. Methods Serum free cells suspension culture and single cell cloning technique were used to isolate neuroepithelial stem cells. 5 bromo 2 deoxyuridine(BrdU) to label new cells combined with single or double immunocytochemistry staining to detect nestin antigen before differentiation and neural cell specific antigens after differentiation, such as neurofilament (NFM 160?kD), glial fibrillary acidic protein(GFAP), galactocerebroside(GalC) and tyrosine hydroxylase(TH). Striatal extracts were used to induce neuroepithelial stem cells to differentiate into dopaminergic neurons. Results The cells isolated from the spinal cord neural tube of the embryonic rat expressed nestin antigen. They had the potential to serially passage and differentiate into neurons, astrocytes and oligodendrocytes. Striatal extracts could induce 12% of them to differentiate into dopaminergic neurons compared with 3% in controls.Conclusion The cells, which express nestin antigen, isolated from neural tube are multipotent and have the ability to self renew, therefore, they are neural stem cells. These stem cells can be induced to differentiate into specific neurons in vitro. Which can provide materials for neural transplantation.
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Objective To observe cell proliferation and neurogenesis in the pregnant mouse dentate gyrus (DG) and the subventricular zone(SVZ). Methods Injection of the thymidine analog 5 bromo 2 deoxyuridine (BrdU)to determine the extent of cell proliferation combined with single or double immunohistochemistry staining with antibodies BrdU,TuJ1(class Ⅲ ? tubulin,neuron specific early differentiation marker)and GFAP. Results The number of BrdU positive cells in the pregnant mouse dentate gyrus was significantly more than that of unpregnant mouse dendate gyrus but not in the subventricular zone.In dentate gyrus,approximately 80% of these cells were neuronal characteristics (TuJ1 immunoreactive)and 3%～5% of these cells were astrocytic characteristics(GFAP immunoreactive).Conclusion\ These findings suggest that pregnancy significantly increases cell proliferation and neurogenesis in the adult mouse dentate gyrus and present the possibility that these new cells exert an important influence on hippocampal function