[Objective]To observe the growth pattern of oligodendrocyte precursor cells(OPCs) in the primary culture from neonatal rat cerebral cortices and study the methods of cell culture and isolation in order to obtain purified OPCs for the experiments of cell transplantation.[Method]The mixed glial cells from the cerebral cortices of 48-hour-old Sprague-Dawley(SD) rats were cultured in vitro.Until 9-10 days in vitro OPCs were isolated and purified by the shaking process and differential adhesion,and then continued OPCs culture in the defined medium.The growth pattern of OPCs in vitro was investigated by contrast phase microscopy and OPC s were further identified with the immunocytochemical techniques.[Result]The distinct stratification of OPCs and astrocytes developed around 9-10 days in primary culture.At this point,the OPCs scattered on the top of the monolayer astroctyes and the soma of most OPCs typically appeared oval or round with two or three processes.The purity of the isolated OPCs reached 95% and these OPCs further developed into the mature oligdendrocytes which were immunoreactive to the specific antigen of oligodendrocyte lineage cells,Oligo2.[Conclusion]OPCs separated from the cerebral cortices of neonatal SD rats can be maintained as immature precursor cells in culture,and OPCs are able to be purified by shaking and differential adhesion under the condition of appropriate cell stratification.

Objective To investigate the change of immunologic function of red blood cell (RBC) in patients with chronic gastritis. Methods The immunologic function of RBC in 88 cases of chronic gastritis was dynamically detected with yeast rosette method. Results The results showed that RBC-C3b receptor rosette (RBC-C3b-RR) rate and RBC immune adherence enhance factor (RFER) rate were significantly lower (P＜0.05) and RBC immune complex rosette (RBC-ICR) rate and RBC immune adherence inhibitor factor (RFIR) were significantly higher (P＜0.05) in the patients with atrophic gastritis than those in control group. In the patients with superficial gastritis, the difference had no distinct significance (P＞0.05). Conclusion The immunologic function of red blood cell (RBC)measurement plays an important role in differential diagnosis and prognosis evaluation of chronic gastritis.

Objective To investigate the effects of transplantation of oligodendrocyte precursor cells (OPCs) on axonal myelination after spinal cord injury in a rat model. Methods A rat model of spinal cord injury at the tenth thoracic vertebral level (T10) was produced by Allen weight-drop impact method. OPCs implantation was performed at the subacute stage of spinal cord injury. Effects of OPCs transplantation on axonal myelination after spinal cord injury were evaluated by HE staining, immunohistochemistry, myelin staining and transmission electron microscopy. Results The implanted cells were still observed in lesioned segments of spinal cord eight weeks after transplantation. The results of HE staining clearly showed better structure of spinal cord in OPCs-transplanted group than that of control group.Myelin staining also demonstrated that the amount of myelin in white matter of lesioned cord in the OPCs-transplanted group (7 802.42 ± 1085.58) was higher than that of the control group (5 055.98 ± 916.74)(P ＜0.01 ). Expression of myelin basic protein (MBP) was significantly increased in the OPCs-trans-planted group (8 544.44 ±812.78) as compared with that of the control group (5 243.83 ±808.27)(P＜0.01). Moreover, transmission electron microscopy further confirmed the improvement of micro-structure of myelination in OPCs-treated rats. Conclusion OPCs transplantation can improve axonal myelination in rat with spinal cord injury.

Objective To prepare the acellular scaffold of spinal cord and analyze its component.Methods The acellular scaffold was prepared with the freeze thawing and chemical extraction,its structure was observed by HE and SEM,its component was analyzed by immunohistochemistry.Results The cells,myelin sheath and axon of nerve fibers in the rat spinal cord were eliminated,but three-dimensional supports of extracellular matrix were reserved.The analytical results showed the component of the acellular spinal cord contain laminin,fibronectin and type Ⅳ collagen—the necessary components to facilitate and induce the regeneration of the injured nerves and enhance the adhesion and proliferation of cells.Conclusion The acellular spinal cord has three dimensional structure and contains several proteins related to the regeneration of the injured nerves and promotion of the survival and proliferation of cells.

Objective To investigate the effects of inhibiting ubiquitin proteasome pathway(UPP) on proliferation of gastric carcinoma cells and the possible mechanism was discussed. Methods The gastric carcinoma cell strain SGC 7901 was treated with MG 132 to inhibit its UPP specially. The effect of growth suppression on cells was evaluated with MTT assay. Cell cycle and apoptosis were detected by flow cytometry(FCM). DNA fragment analysis was used for confirming the presence of apoptosis. The activity of telomerase was examined by TRAP PCR ELISA. Expression of p27kip1 was detected by immunocytochemical technique. Results MG 132 had great inhibitory effect on the growth of SGC 7901 cells. The FCM analysis showed that the ratio of G0/G1 phase of control group was (46.3?4.1)%, the ratio of G0/G1 phase of SGC 7901 cells treated with MG 132 increased to (72.1?5.0)% ( P

Objective To investigate the role of cytokines genes expression of liver sinusoidal endothelial cells in liver damage by sepsis.Methods Septic mice models were established with cecal ligation and perforation (CLP), while sham operation group received the same procedure exclusive CLP. The genes expression of TNF?, IL 1? and IL 6 in liver sinusoidal endothelial cells were assessed by RT PCR. Results A significant increase of TNF?, IL 1? genes expression was observed at 3h, and a slight decline at 12h after operation, but still significantly higher than that in the sham group; while IL 6 gene expression showed signficantly higher at 3h and remained at the high level at 12h. Conclusions Liver sinusoidal endothelial cell is an important source of cytokine production in mice with sepsis.

ObjectiveTo optimize the condition of multiple fluorescence quantitative PCR and establish a new assay of four human herpes virus (HHV) detected by AllGlo quadruple fluorescence quantitative PCR.MethodsFour HHV including HSV-1,HSV-2,EBV and CMV were identified by sequence analysing the qualitative PCR production.Furthermore,they were quantitatively detected by AllGlo and TaqMan multiple fluorescence quantitative PCR respectively.ResultsBoth the positive rate and specificity of AllGlo and TaqMan in detecting single HHV achieved 100％.And AllGlo single fluorescence quantitative PCR prevailed over TaqMan's by Ct of 1-3.Four HHV can be simultaneously detected by AllGlo quadruple fluorescence quantitative PCR,comparing to the only two by TaqMan.ConclusionAllGlo fluorescence quantitative PCR assay allows a higher throughput,sensitivity and specificity than TaqMan in detection and thus provides a board prospect.

Objective To evaluate the application of fluorescence PCR in detecting ureA gene of Helicobacter pylori(HP)in feces.Methods Fluorescence PCR was used to detect ureA gene of HP in feces from 50 patients,including 23 confirmed by gastric biopsy urease test and histological staining.Bacterium culture and serum antibody detection were also performed, and chi-square test was used to compare the sensitivity,specificity,positive predictive value and negative predictive value among three methods.Results The sensitivity,specificity,positive predictive value and negative predictive value of fluorescence PCR were 1.00,0.96,96.O%and 100.0%,while those for HP culture were 0.78,1.00,100.0%and 84.0%,and thee for serum antibody detection Was 0.96,0.74,76.O%and 95.0%.There were significant differences in sensitivity and negative predictive value between PCR and bacterium culture (X2=5.60 and 4.44,P<0.05),and significant differences in specificity and positive predictive value between PCR and serum antibody detection(X2=5.28 and 4.08,P<0.05).Conclusion ureA gene detection in feces by fluorescence PCR is of value for the diagnosis of HP infection.

Objective To explore clinical effect of metacentric particles with bisoprolol fumarate pesos on the elderly patients with heart failure with ventricular arrhythmia treatment.Methods In August 2013 to April 2016, 102 cases of elderly patients in our hospital heart failure with ventricular arrhythmia were selected as research object, according to touch the ball method, they were divided into control group and experimental group, each group of 51 cases, control group was given bisoprolol fumarate pesos, on this basis, the experimental group was combined use of metacentric particles.Two groups of clinical curative effect were compared .Results The experimental group and control group rate of sinus heart rate to maintain statistical comparison of the experimental group was higher than control group ,the difference was statistically significant(P<0.05),group total effective rate was significantly higher than the control group ( P <0.05 ) , the incidence of adverse reactions were significantly lower than the control group (P<0.05),the control group and experimental group heart rate compared before treatment, there was no statistically significant difference, but after treatment, the experimental group was lower than the control group, the differe nce was statistically significant (P <0.05). Conclusion Metacentric particles with bisoprolol fumarate pesos for elderly patients with heart failure with ventricular arrhythmia in clinical curative effect is distinct, high safety.

Objective To investigate the expression of 5-lipoxygenase activating protein gene (ALOX5AP) polymorphism in the patients with cerebral infarction,and explore its relationship with cerebral infarction susceptibility.Methods Patients with cerebral infarction and healthy volunteers were selected for this study,whose venous blood was extracted and detected with polymerase chain reaction/restriction fragment length polymorphism (PCR-RFLP).Haplotype A (SG13S114T,SG13S89G,SG13S32A,SG13S25G),haplotype B (SG13S377A,SG13S114A,SG13S41A,SG13S35G),and their nucleotide polymorphism loci were observed.Results Single nucleotide polymorphism (SNP)-SG13S114,SNP-SG13S32 and HapA carrying rate were significantly different between patients with cerebral infarction and healthy volunteers (P ＜0.05).SNP-SG13S114 and SNP-SG13S32 were independent risk factors of cerebral infarction (OR ＞ 1.0,P ＜ 0.05).Conclusions The morbidity of cerebral infarction in Wenling City was influenced by SNP-SG13S114,SNP-SG13S32,and HapA carrying rate.