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

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

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

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.

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.

OBJECTIVETo determine the distribution of HCV genotypes among volunteer blood donors in Guangzhou.

METHODSSix-nine HCV RNA-positive samples were collected from volunteer blood donors in Guangzhou. NS5B fragments of HCV were amplified followed by DNA sequencing and phylogenetic analysis.

RESULTSHCV genotypes were determined for 67 samples. Among them, the subtypes 1b, 2a, 3a, 3b, 6a and 6n were detected at the frequencies of 37.31%, 4.48%, 7.46%, 4.48%, 44.78% and 1.49%, respectively.

CONCLUSIONHCV 1b and 6a are the most predominant two subtypes among volunteer blood donors in Guangzhou.

Blood Donors ; China ; Genotype ; Hepacivirus ; classification ; genetics ; isolation & purification ; Humans ; Phylogeny ; RNA, Viral ; genetics ; Sequence Analysis, DNA

This study was aimed to investigate the characteristics of human bone marrow mesenchymal stem cells during ex-vivo expansion, MSCs were isolated from human bone marrow. At each passage, the characteristics of proliferation kinetics, osteogenic and adipogenic differentiation potential were analyzed, and cell morphology, surface markers were investigated as well. The karyotype analysis was done in different passage cells. The infection HIV, HCV, HBV and TP were detected by ELISA. Mycoplasma contamination in vitro was detected by PCR method. HLA-SBT was used to reanalyze the results of HLA antigens and alleles. STR genetic loci were detected by PCR in the MSC1, MSC2, MSC3 and MSC4. The results indicated that the proliferative ability and osteogenic potential decreased with the increase of passage number during culture expansion. The multiple differentiation potential of MSCs was maintained during their life span. Karyotype analysis showed that MSCs from 4 groups before passage 8 were normal. The expression of CD29, CD44, CD105, CD166 and CD73 were positive. The expression of CD14, CD34, CD45, CD80, CD86 were all negative. SBT was used to identify HLA-A, B, Cw, DRB1, DRPB1, DQ alleles in the MSC1, MSC2, MSC3, MSC4. The genetype of STR in the MSC1, MSC2, MSC3, MSC4 was different. MSC 3 was examined by TP-ELISA to confirm the infectious disease of TP. MSC2 was contaminated by mycoplasma at passage 5. It is concluded that culture expansion causes MSCs to gradually lose their stem cell properties. During ex-vivo expansion of MSCs, the osteogenic differentiation potential is decreased. MSCs before passage 8 can be a valuable subject for basic research and clinical application.
Adipogenesis ; Adult ; Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cells, Cultured ; Female ; Humans ; Karyotyping ; Male ; Mesenchymal Stromal Cells ; cytology ; Osteogenesis

OBJECTIVETo study on the efficacy, prognosis and security of high-intensity focused ultrasound (HIFU) combined with transcatheter arterial chemoembolization (TACE) in the treatment of hepatocellular carcinoma (HCC).

METHODSTotally 72 HCC patients treated by HIFU from December 2009 to January 2011 were divided into two groups according to treatment methods: 40 cases in HIFU group, 32 cases in TACE + HIFU treatment group (combined group). Then set up a control group include 40 cases treated by only TACE in the same period (TACE group). The improvement of clinical symptoms, AFP, reduce rate of tumor volume, survival rate of 1 year after operation and postoperative complications in front and behind the treatment were analyzed.

RESULTSThere was no significant statistical difference on the improvement of clinical symptoms in all these three groups (P > 0.05) after treatment for HCC. There is no significant statistical difference also on reduce rate of tumor volume and decrease rate of AFP in both HIFU group (35.0%, 41.4%) and TACE group (37.5%, 41.9%) (χ² = 0.054, P = 0.816; χ² = 0.002, P = 0.965). Both reduce rate of tumor volume (62.5%) and decrease rate of AFP (72.0%) in combined group were better than HIFU group (χ² = 5.394, P = 0.020; χ² = 5.098, P = 0.024) and TACE group (37.5%, 41.9%) (χ² = 4.448, P = 0.035; χ² = 5.062, P = 0.024). Kaplan-Meier survival curve showed that there was no significant statistical difference on short-term survival rate in the 3 groups. But the long-term survival rate of combined group was better than TACE group and HIFU group.

CONCLUSIONTACE combined with HIFU is a effective, safe and noninvasive treatment method to HCC.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; therapy ; Chemoembolization, Therapeutic ; Combined Modality Therapy ; Female ; Follow-Up Studies ; Humans ; Liver Neoplasms ; therapy ; Male ; Middle Aged ; Prognosis ; Treatment Outcome ; Ultrasound, High-Intensity Focused, Transrectal

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

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.