Objective To study the neuroprotective effect of erythropoietin(EPO) on neonatal rats model with hypoxia-ischemia encephalopathy(HIE).Methods HIE was induced in rats on 7th day of postnatal age by ligation of right common carotid artery,followed by 2 h of hypoxia(80 mL/L O2).The subjects were divided into sham-operated group,control group and EPO group.EPO 4 000 U/(kg?day) was injected daily from day 2 pre-surgery for 9 to 16 days and PBS was injected in the control group.The neuroprotective effect of EPO on HIE model was detected by brain weight,the difference in weights between the ipsilateral(right) and contralateral(left) brain and the function test.In vitro study,the neural progenitor cell line C17.2 under gone apoptosis following an ischemia-like metabolic inhibition.The effect of EPO on the cell line ischemia modle 17.2 was evaluated by detecting Annexin V with flow cytometry.Results The signi-ficant and sustained brain injury in the hypoxia-ischemia and vehicle-treated group was observed and measured by reduction in relative weights of ipsilateral to contralateral and compromised sensorimotor functions in response to postural reflex test,compared with those of sham-operated animals(Pa

Objective To study the correlation between fractional anisotropy(FA)and tumor microarchitecture(MVD,VEGF and celluarity).Methods Fouteen gliomas(5 grade Ⅰ and Ⅱ,4 grade Ⅲ, 5 grade Ⅳ)confirmed histo-pathologically were performed on diffusion tensor imaging(DTI)using a GE Signa Excite Ⅱ 3.0 T MR scanner(8-channel head coil,SE echo planner imaging(EPI),thickness:5 mm, spacing:0,directions:25,B values:0 and 1000 s/mm~2,TR 6000 ms,TE minimum,FOV:240 mm? 240 mm,image matrix 128?128,NEX 2).Postprocessing was done using a DTI specific software to gain FA image.ROIs were drqwn in tumor parenchyma and the value of FA was recorded.The positive expression of VEGF and CD34 was shown using immuno-histochemistry method.The VEGF,MVD,and cellularity of every slices were recorded.Pearson correlation analysis was used.Results FA(which is 0.102?0.080 in grade Ⅰ and Ⅱ,0.171?0.037 in grade Ⅲ,0.200?0.021 in grade Ⅳ)has the trend to raise with the increasing grade of astrocytomas.FA has significant positive correlation to MVD(40/HP in grade Ⅰ and Ⅱ, 86/HP in grade Ⅲ,101/HP in grade Ⅳ),VEGF(8% in grade Ⅰ and Ⅱ,47% in grade Ⅲ,55% in grade Ⅳ),and cellularity(104/HP in grade Ⅰ and Ⅱ,160/HP in grade Ⅲ,265/HP in grade Ⅳ).The correlation coefficients between FA and VEGF,MVD,and cellularity were 0.748,0.668,0.625 respectively.Conclusion As a new imaging method,DTI can reveal the microarchitecture in gliomas and be value of distinguishing gliomas of different grade.DTI provides a new method of precise diagnosis to glioma preoperatively.

OBJECTIVETo investigate the expression and clinical significance of Ephrin-A1 mRNA and protein expression level in hepatocellular carcinoma.

METHODSReverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry technique were used to detect the expression of the Ephrin-A1 mRNA and protein of 40 cases of hepatocellular carcinoma and their corresponding para-cancerous tissues and 10 cases of normal liver tissues. The relationships with its clinical pathology characters were analyzed.

RESULTSThe mRNA of Ephrin-A1 was expressed in all of the 40 cases of hepatocellular carcinoma and their corresponding para-cancerous tissues and 10 cases of normal liver tissues. Semiquantitative analysis showed that the mRNA expression level of Ephrin-A1 in hepatocellular carcinoma (0.5413 +/- 0.1527) was greater than that in corresponding para-cancerous tissues (0.3895 +/- 0.0549, P < 0.05) and normal liver tissues (0.3770 +/- 0.1055, P < 0.05); but between corresponding para-cancerous tissues (0.3895 +/- 0.0549) and normal liver tissues (0.3770 +/- 0.1055), the mRNA expression level had no significant difference (P > 0.05). The positive rates of Ephrin-A1 protein were 20% (2/10) in normal tissues, 35% (14/40) in para-cancerous tissues and 62% (25/40) in hepatocellular carcinoma tissues, respectively; the protein expression level of Ephrin-A1 was gradually rising (chi(2) = 14.762, P < 0.05). The overexpression of Ephrin-A1 protein was correlated with histological differentiation, tumor thrombi in portal vein and lymph node metastasis (P < 0.05).

CONCLUSIONSThe overexpression of Ephrin-A1 protein is correlated with histological differentiation, the lymph node metastasis and tumor thrombi in portal vein. It indicates that Ephrin-A1 may play an important role in the malignancy transformation, invasion progression and metastasis of hepatocellular carcinoma.

Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Differentiation ; Ephrin-A1 ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Neoplasm Metastasis ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the expression and clinical significance of EphA7 protein in primary hepatocellular carcinoma.

METHODSImmunohistochemistry and Western blot were used to detect the expression of EphA7 protein in 40 cases of primary hepatocellular carcinoma, their corresponding adjacent liver tissues and 10 cases of normal liver tissues. The relations with its clinical pathological parameters were analyzed too.

RESULTSExpression of EphA7 protein was mainly located in the cytoplasm and the blood vessels of the septa, which was found in hepatocellular carcinoma tissues, their corresponding adjacent liver tissues and normal liver tissues. Western blot analysis showed that the expression level of EphA7 protein in hepatocellular carcinoma (0.58 +/- 0.26) was greater than that in corresponding adjacent liver tissues (0.40 +/- 0.22, P < 0.05) and normal liver tissues (0.32 +/- 0.16, P < 0.05). But it had no significant difference between corresponding adjacent liver tissues and normal liver tissues (P > 0.05). EphA7 protein expression was correlated with histological differentiation, tumor thrombi in portal vein, lymph node metastasis and high AFP level (P < 0.05).

CONCLUSIONSEphA7 protein expression is significantly correlated with the biological behavior of primary hepatocellular carcinoma. The high expression of EphA7 protein may play an important role in the malignancy transformation, invasion progression and metastasis of primary hepatocellular carcinoma.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Receptor, EphA7 ; metabolism

Objective To study the inhibitory effects of troglitazone on the proliferation of rat pituitary adenoma GH3 cell line in vitro and explore the mechanisms. Methods GH3 cells were treated with troglitazone at different concentrations (1×10-7, 1×10-6and 1×10-5 mol/L), dimethyl sulfoxide (DMSO) (DMSO control group) or phenol red- and serum-free F-12 medium (blank control group). MTT assay and flow cytometry was used to detect the cell growth and the cell cycle distribution after the treatment, respectively. Semi-quantitative RT-PCR was performed to detect the expression of cyclin D1 mRNA. Results Troglitazone treatment for 72 h significantly inhibited the cell proliferation and induced obvious G1/S cell cycle arrest and cell death. Compared to those in the blank control and DMSO-treated cells, troglitazone also significantly decreased the expression ofcyclin 1I mR_NA in the GH3 cells in a concentration-dependent manner (P<0.05). Conclusion Troglitazone can obviously inhibit the proliferation of GH3 cells possibly through the mechanism of decreasing cyclin D1 mRNA after its binding to peroxisome proliferator-activated receptor-γ, which induces G1 cell-cycle arrest and promotes cell death.

Objective:To investigate the HBV infection and analyze the related risk factors among Li minority in Baisha county,Hainan Province. Methods: A total of 1 595 individuals of Li minority in Baisha county,were enrolled by random sampling method from July 2014 to October 2015. Epidemiological data including baseline characteristics and risk factors were obtained. HBcAb was detected by chemiluminescence method. The difference in age between HBcAb positive and negative group was analyzed by t test. The effects of age,gender and related risk factors on HBcAb were analyzed by univariate chi-square test and multivariate Logistic regression. Results: The positive rate of HBcAb was 71. 8% (1 145/1 595) and no significant difference between male and female was observed(x2=0. 134,P=0. 715). The difference of HBV infection among age groups was statistically significant (F=540. 769,P＜0. 001). The HBV infection rate was 11. 9% in the 12-17 year group,which was significantly lower than the others. The rate in the 18-23 year group (28. 0% ) was significantly higher than that in the 12-17 year group,but significantly lower than the other groups (＞85% ). Multivariate Logistic regression analysis showed that alcohol consumption and tattoo were independent risk factors associated with HBV infection (x2=169. 833,P＜0. 001;x2=11. 354,P=0. 001). Conclusion: The rate of HBV infection of Li minority in Baisha county,Hainan Province is high. The age,alcohol consumption and tattoo are the independent risk factors for infection.

Based on standard carotid endarterectomy, we performed modified carotid endarterectomy in two cases of carotid artery stenosis by changing the direction of the carotid artery incision to avoid restenosis of the internal carotid artery without using a patch. The two patients recovered smoothly without any complications. Compared with eversion or patch endarterectomy, this modified carotid endarterectomy avoids restenosis of the carotid artery and shortens operation time.

We investigated the molecular characteristics of the full-length genome of 5 dengue serotype 4 virus (DENV-4) strains isolated in Yunnan Province,China,2015 and their molecular epidemiological feature.Isolation of dengue virus was using C6/36 cell culture method.Viral RNA was extracted from virus isolates,then the full-length genome was amplified by RT-PCR.The homology and phylogenetic analysis was made on the nucleotide and deduced amino acid sequences by bioinfor matics softwares including ClastalX1.83 and MEGA6 etc.Results showed that five strains of DENV-4 isolated from dengue fe ver cases in Ruili City of Yunnan Province in 2015,of these,2 strains from indigenous cases,3 from imported cases from Lashio and Nam Can cities of Myanmar to Ruili of China.RT-PCR and sequencing indicated that the full-length genes (10 661 nt) of 5 DENV-4 strains were obtained,and their open reading frame (103-10 264) were coded 3 386 amino acid residues.Phylogenetic tree and homology analysis based on the comeplete genome or structural and non-structural protein genes showed that the 5 DENV 4 isolates were highly homologous and gathered in an evolution as well as they have a closer genetic relationship with the DENV-4 genotype Ⅰ (G-Ⅰ) strains isolated from Thailand.Results indicated that the Yunnan strains belonged to G-Ⅰ.Yunnan strains and Thailand strains compared with DENV 4 prototype strain (H241,Philippines 1956) and Guangzhou strain (B5,1990) of China and showed low homology and evolutionary relationship.When Yunnan strains compared with H241 strain,there were 21 and 45 different sites in amino acid of structural and non-structural proteins,respectively.This is the first time in Yunnan to obtain full-length genomes sequence of DENV-4 and they have closer evolutionary relationship with DENV 4G-Ⅰ strains from Southeast Asia region in recent years.The autochthonous DENV-4 epidemic in Yunnan was detected for the first time,and the virus transmission sources were from neighboring northern Myanmar.It is necessary to further study that change of the amino acid sites of Yunnan strains of DENV-4 is related to its antigenicity and virulence.

OBJECTIVETo investigate the effect of human platelet lysates (HPL) obtained from platelet-rich plasma on the proliferation and biological characteristics of human mesenchymal stem cells (MSCs) in vitro.

METHODSHPL was obtained by repeated freeze-thawing of human plateletes, and the MSCs separated by density gradient centrifugation from 6 donors were expanded in medium supplemented with 10% fetal bovine serum (FCS) or HPL at different concentrations. The optimal concentration of HPL for cells culture was determined according to the cell proliferation kinetics. The cultured MSCs were characterized for their proliferation, cell phenotype, and cell cycle distribution.

RESULTSThe HPL-supplemented medium contained 4 essential growth factors for the growth of MSCs, namely platelet-derived growth factors (0.53∓0.06 ng/ml), basic fibroblast growth factor (37.5∓4.31 pg/ml), insulin-like growth factor-1 (0.15∓0.06 mg/ml) and transforming growth factor (5150∓463 pg/ml). Cultured in the presence of HPL at the optimal concentration of 7.5%, the MSCs displayed a spindle-shaped fibroblast-like morphology without obvious changes in the proliferation activity till passage 8 (P>0.05), similar to those of cells in FCS-supplemented culture medium. Flow cytometry and cell cycle analysis revealed no differences in the phenotypes or cell cycle distribution between the cells cultured in the presence of 7.5% HPL and 10% FCS.

CONCLUSIONThe culture medium supplemented by 7.5% HPL can promote the expansion of human MSCs and maintain the basic biological characteristics of the cells.

Blood Platelets ; cytology ; metabolism ; Cell Extracts ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media ; pharmacology ; Fibroblast Growth Factor 2 ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Platelet-Derived Growth Factor ; pharmacology

OBJECTIVETo investigate the correlation between the platelet GP specific antibodies/HLA antibodies and platelet transfusion refractoriness (PTR).

METHODSSixty-five patients with PTR were selected in this study and were genotyped for HLA-A and HLA-B as well as HPA systems by standard PCR-SSP assays. The platelet GP specific antibodies and HLA antibodies in serum and platelet elution were tested with a solid phase ELISA.

RESULTSThe HLA-A/B antigens and the frequencies of HPA-1, 2, 4, 5, 6, 9, 15 antigens in PTR patients had no difference from those in healthy donors. The freguencies of HPA-3a and 3b were 0.65 and 0.35, respectively. There was statistical difference between the 65 PTR patients and the healthy donors in HPA-3 freguencies (P < 0.05). Twenty-four patients (36.9 %) only expressed HLA antibodies, and 14 (21.5%) expressed HLA and platelet GP specific antibodies. The highest expression of anti-HLA-A/B specific antibodies was -A*9(46.2 %)/-B*40(33.6%), respectively. In serum, GPIIb/IIIa was expressed (26.2%), followed by GPIa/IIa (21.5 %). In platelet elution, GPIIb/IIIa was expressed of 41.5% and GPIb/IX 41.5%. Pedigree study was carried out for 2 patients. The results showed that the platelet GP specific antibody/HLA antibody developed in PTR patients was highly related to the mismatch with the platelet antigen/HLA antigen in their parents.

CONCLUSIONThe expressions of the HLA and platelet GP specific antibodies are the most important reason in PTR, it's meaningful to explore the correlation between PTR and HLA and HLA-A/B antigen in guiding platelet transfusion.

Antigens, Human Platelet ; immunology ; Blood Platelets ; Humans ; Isoantibodies ; immunology ; Platelet Transfusion ; Thrombocytopenia