BACKGROUND:PA6 cells are bone marrow stromal stem cells from the mouse skul , and scientists have found that PA6 cells co-cultured with some kinds of stem cells have shown neural differentiation, based on which, PA6 cells can be used for repair of nerve injury. Therefore, researchers give more and more attentions to PA6, but few studies have addressed isolation and culture methods of bone marrow stromal stem cells from the skul . OBJECTIVE:To isolate and culture bone marrow stromal stem cells from the skul of Sprague-Dawley rats and to observe cellular morpholopy in vitro and perform immunofluorescence identification. METHODS:Under sterile conditions, newborn Sprague-Dawley rat’s skul was cut into pieces. Smal skul pieces were washed using Dulbecco’s modified Eagle’s medium containing 10%fetal bovine serum. Single cellsuspension was made, and placed in culture flask. The culture medium was changed many times for cellpurification. The second passage of cells was obtained for morphology observation under an inverted microscope. cellsurface markers were detected by using immunofluorescence staining. RESULTS AND CONCLUSION:After the primary culture for 24 hours, cells exhibited adherent growth;after 3 days, cells were increased in number, and presented with irregular shapes, such as polygon, triangle, spindle and flat shape. After passage, cells were uniform in morphology, arranged in radial and bunch patterns, and exhibited strong adherent ability. Some cells grew in cluster, and proliferated faster than primary cells. Immunofluorescence staining showed that cells were positive for CD105, CD73, CD44, CD90, but negative for CD45, D34,CD14, HLA-DR. Results indicate that this method can obtain bone marrow stromal stem cells.

0.05).(4)Differences in Apelin level were found between the NC group and the CHD group both before and after angiography(90.25?2.96 ?g/L vs 77.04?3.89 ?g/L in arterial blood and 87.83?3.34 ?g/L vs 75.14?3.16 ?g/L in venous blood before angiography;91.25?2.04 ?g/L vs 78.11?2.84 ?g/L in arterial blood and 90.79?2.58 ?g/L vs 77.43?2.38 ?g/L in venous blood after angiography;all P values

Objective To explore causes and high risk electrocardiogram expression of cardiac syncope caused by malignant rapid ventricular arrhythmia.Methods Forty-eight patients were analysed, they had cardiac syncope once or more that after which were admitted to the hospital. Results The basic causes of cardiac syncope were individed following types in 48 patients: coronary heart disease with acute or dated myocardial infarction,dilated or hypertrophic cardiomyopathy,hypokalaemia were ordinarist inducer.Torsade de pointes(TdP) were most common type of malignant rapid ventricular arrhythmia.They had some high risk electrocardiogram expression: secondary long QT syndrome,Brugada's syndrome,idiopathic abnormal J wave,complex ventricular ectopic beats,acute myocardial infarction with ST-T electrical alternation; or extensive anterior myocardial infarction with tombstone ST segment elevation,dilated cardiomyopathy with advance QRS complex low voltage.They were different electrocardiographic and clinical characteristic. Conclusion The cardiac syncope caused by malignant rapid ventricular arrhythmia is not single and independent clinical entity, which presents different the causes and high risk electrocardiogram expression.

Objective To compare the distribution and expression differences of glycogen synthase kinase-3?(GSK3?) among normal adult,aged and amyloid beta(A?)-induced neurodegenerative rat brains,so as to explore its functional role in neurodegeneration. Methods Aggregated A? was microinjected into normal adult rat hippocampus under a stereotaxic system. The rats over 12 months were defined as aged rats. The distribution and localization of GSK3? were examined using immunohistochemistry. Western blotting was performed to assess expression change in cortex and hippocampus quantitatively. Results The GSK3? positive cells were distributed extensively around the whole brain and almost with neuron-like morphology. In normal adult rats,the strong anti-GSK3? immunoreactivity located in the neocortex pyramidal layer,hippocampus pyramidal layer,dentate gyrus,thalamus,substantia nigra,etc. The amount of GSK3? positive cells was much more in the aged and A?-injected group than in normal ones. The immunoreactive signals usually extend to the distal area of neurite in the A?-injected ones. Western blot showed that the expression intensity of GSK3? was stronger in the aged and neurodegenerative rat brain than in the normal adult rat brain. Conclusion The expression of GSK3? increases apparently in the neurons of aged and A?-injected brain. It may play a role in the neurodegenerative process.

BACKGROUND:Schwann cells transplantation can change the local micro-environment and help to repair the injured neural tissue, so getting a large number of highly purified and active Schwann cells is the key of the study. OBJECTIVE: To search for a simple and rapid method to extract and purify the Schwann cells.METHODS: Rats were divided randomly into two groups, namely, in vivo pre-degeneration of sciatic nerve resection group and untreated control group, with 20 rats in each group. Under sterile conditions, the rat sciatic nerves were cut off at post-operative 7 days, Schwann cells were extracted by using mixed enzyme digestion and tissue mass transplantation; through low enzyme digestion and twice inoculation to differential adhesion, Schwann cells were purified. Cell morphology was observed under phase contrast microscope and identified by mmunofluorescence staining; cell purity was calculated; MTT method assay was used to determine the capacity of cell proliferation.RESULTS AND CONCLUSION: At 7 days of the culture, the experimental group showed the typical bipolar or bipolar Schwann cells, with connections between cells; in control group, cell processes were shorter and less associated with the surrounding cells. Following S-100 immunofluorescence staining, cells were positive for green expression.Cells proliferated rapidly in the experimental group and formed a swirling shape at 15 days, there were a relatively small number of fibroblasts, at the purity of 96.1%; in the control group, the cells proliferated slowly, with many fibroblasts at a low purity. MTT assay showed that primary cultured Schwann cell proliferated weakly in both groups; compared with the control group, the proliferation of subcultured Schwann cells in the experimental group was markedly increased (P < 0.05 or 0.01), and reached a peak 3 4 days later. The results confirmed that in vivo denaturing, in vitro hybrid enzyme digestion, tissue mass transplantation combined with low enzyme digestion, separation of double-differential adhesion of Schwann cells is a simple and rapid method to extract and purify Schwann cells.

Objective To investigate the effect of lithium chloride(LiCl),an inhibitor of glycogen synthase kinase-3beta(GSK-3beta),on proliferation and differentiation of neural stem cells(NSCs).Methods The NSCs were isolated from cortex of rat fetus and expanded in culturing system.Their morphological changes and attachment process were observed under microscope.The cell cycle dynamics of NSCs was examined with flow cytometry.And the expression of GSK-3β and β-catenin was examined quantitatively with Western blot.Results The culturing NSCs treated with LiCl were usually floated and much dispersed in the media.Many of the neurospheres became small and the time of attachment after serum induction became longer.Using flow cytometry,it was detected that the proportion of G1 phase NSCs declined gradually accompanying the increased concentration of LiCl,while the percentage of S and G2/M phase cells showed an increasing trend.Western blotting results revealed β-catenin expression increased whereas Gsk-3βdecreased gradually under the treatment of LiCl and also showed a dose dependent manner.Conclusion These results suggest that LiCl may promote the proliferation of NSCs and prevent them from differentiating,which may partly involve the activation of wnt/β-catenin signaling pathway.

BACKGROUND:Studies have shown that Schwann cells form a Bunger band in the basement tube and guide the extension of regenerating axons after peripheral nerve injury, but the exact mechanism remains to be explored.
OBJECTIVE:To explore the effect of Wal erian degeneration on biological characteristics and secretory function of Schwann cells in rats with sciatic nerve injury.
METHODS:A rat model of sciatic nerve injury was established and divided into two groups:sciatic nerve transection group and surgical control group. Schwann cells were isolated and cultured from sciatic nerve segments by one enzyme digestion. The cellmorphology was observed under light microscope and S-100 protein expression was determined by immunofluorescence staining. After subculture, the first generation of Schwann cells were chosen to draw the growth curve by the counting method within 14 days. The cellactivity was detected by MTT assay. The adhesion of Schwann cells was examined by acid phosphatase analysis and the concentration of nerve growth factor was detected by ELISA method.
RESULTS AND CONCLUSION:At 14 days after primary culture, a great number of Schwann cells were observed near the edges of nerve segments in the sciatic nerve transection group, but only smal number of Schwann cells scattered around nerve segments in the control group. Schwann cells in both groups showed S-100 positive expression. At 3 days after subculture, Schwann cells reached the logarithm proliferative phase, the cellnumber and proliferation absorbance values in both groups were increased along with time extension. Furthermore, the number of Schwann cells and absorbance value in the sciatic nerve transection group were significantly higher than those of control group (P<0.05). The adhesion ability in the sciatic nerve transection group was also significantly higher than those in the control group (P<0.05). ELISA results showed that, the concentrations of nerve growth factor in the sciatic nerve transection group were significantly higher than those in the control group at 4, 6, 8, 10, 12 and 14 days (P<0.05). After sciatic nerve injury, Wal erian degeneration can induce Schwann cells dedifferentiate into the precursors, significantly influence the biological function of Schwann cells, promote the proliferation of Schwann cells within the short term, secrete large amounts of neurotrophic factors, enhance celladhesion, and provide a suitable microenvironment for regenerated axons. In addition, it creates the necessary microenvironment for peripheral nerve regeneration.

Objective To establish the Cookie Theft Test for Chinese norms. Methods 29 normal participants and 17 patients with stroke finished the Cookie Theft Test. The language samples were analyzed in 7 different indexes which are incorrect statement (IS), dysfluencies (DF), providing structure support (PS), repetitions (RP), content units (CU), elaborations of content units (EC) and irrelevancies (IR). Besides, the total words of language samples were counted. The scores of normal participants in 8 indexes were regarded as norms. And the scores of patients in 8 indexes were used to test the validity of the norms. Results The test-retest reliability was r(IS)=0.92, r(DF)=0.89, r(PS)= 0.98, r(RP)=0.84, r(CU)=0.96, r(EC)=0.88 and r(IR)=0.99, respectively. 12 out of 17 patients were distinguished by the norms. Conclusion The norms of Cookie Theft have acceptable reliability and validity and can be applied to clinical diagnoses and scientific researches

A simple and rapid immunochromatographic test strip incorporating a colloidal gold-labeled recombinant Nsp7 antigen probe was successfully developed for the detection of anti-porcine reproductive and respiratory syndrome virus (PRRSV) antibodies in swine. Recombinant Nsp7 protein of PRRSV labeled with colloidal gold was dispensed on a conjugate pad for use as the detector. Staphylococcal protein A and purified porcine anti-Nsp7 antibodies were blotted on a nitrocellulose membrane to form test and control lines, respectively. A comparison of the strip with standard diagnostic tests, enzyme-linked immunosorbent assays and immunoperoxidase monolayer assay, was also performed. The immunochromatographic test strip was shown to be of high specificity and sensitivity. Furthermore, the strip assay is rapid and easy to perform with no requirement for professional-level skills or equipment. It is suggested that the immunochromatographic test strip can be used to quickly and accurately detect PRRSV antibody and to be suitable for diagnostic purposes in the field.
Antibodies* ; Collodion ; Colloids ; Diagnostic Tests, Routine ; Enzyme-Linked Immunosorbent Assay ; Gold Colloid ; Immunochromatography ; Membranes ; Porcine Reproductive and Respiratory Syndrome* ; Porcine respiratory and reproductive syndrome virus* ; Sensitivity and Specificity ; Staphylococcal Protein A ; Swine

Objective:To explore the value of MR elastography (MRE) and shear wave elastography (SWE) for staging liver fibrosis in a rabbit model.Methods:From March to November 2020, 200 healthy New Zealand white rabbits were randomly divided into control group ( n=40) and liver fibrosis group ( n=160) by random number table method. The volume ratio of CCl 4 and olive oil was 1∶1 to prepare 50% CCl 4 oil solution, and the experimental rabbits in the liver fibrosis group were subcutaneously injected with 50% CCl 4 olive oil solution. It was injected once a week at the dose of 0.1 ml/kg in the first to third weeks, once a week at the dose of 0.2 ml/kg in the 4th to 6th weeks. The dose of 0.1 ml/kg was injected twice a week from week 7 to 16. The control group were subcutaneously injected with an equal dose of normal saline. At the end of the 4th, 8th, 12th, and 16th week, 40 and 10 animals in the liver fibrosis group and the control group were randomly selected by random number table method for MRE and SWE, respectively, to obtain the liver elastic stiffness (LS), which were recorded as LS MRE and LS SWE. After the examination, the experimental rabbits were sacrificed and liver tissue of rabbits were taken for histopathological Scheuer staging, and they were divided into F0-F4 groups. One-way ANOVA was used to evaluate the differences of LS MRE and LS SWE in different stages of liver fibrosis. Spearman correlation was used to analyze the correlation between LS and pathological stages. The receiver operating characteristic curve was used to evaluate the efficacy of LS MRE and LS SWE in diagnosing liver fibrosis staging, and the area under the curve (AUC) was compared using the Z test. Results:Totally 162 rabbits were included, which covered F0 ( n=38), F1 ( n=33), F2 ( n=35), F3 ( n=31) and F4 ( n=25). Significant differences of LS MRE and LS SWE values were found among different stages of liver fibrosis ( F=295.29, 102.40, both P<0.001). LS MRE, LS SWE were both positively correlated with liver fibrosis stage ( r=0.93, 0.81, both P<0.001). The AUC of LS MRE for diagnosing liver fibrosis stages ≥F1, ≥F2, ≥F3, and ≥F4 were 0.955, 0.967, 0.996, and 0.980, respectively; the AUC of LS SWE were 0.856, 0.880, 0.974, and 0.953, respectively. The AUC of liver fibrosis stage ≥ F1, ≥ F2 for LS MRE value were greater than LS SWE value ( Z=2.93, 3.29, P=0.003, 0.001), and the AUC of ≥F3, ≥F4 had no significant differences ( Z=1.58, 1.68, P=0.115, 0.093). Conclusion:Both MRE and SWE can accurately predict the stage of liver fibrosis in experimental rabbits, and MRE is better than SWE in diagnosing early liver fibrosis.