OBJECTIVETo investigate the effect of local administration of recombinant adenovirus of human adiponectin (hAPN) Ad-hAPN-EGFP on tibial defect repair of SD rats.

METHODSTibial defect (2 mm) models of 36 SD rats (72 sides) were randomly divided into three groups (A, B, and C; n=24). The three groups were injected with Ad-hAPN-EGFP, Ad-EGFP, and normal saline, respectively, during operation and the day after operation. A week after the operation, the expression of hAPN and osteogenesis-related factors were detected by real-time polymerase chain reaction. Three weeks after operation, the tibias were examined by micro-computed tomography, hematoxylin-eosin staining, and Masson staining to evaluate the restoration of bone defects.

RESULTS1) The hAPN expression was detected in group A but not in groups B and C. Osteogenesis-related factors expression of group A was significantly higher than that of the other groups (P<0.05). 2) Osteogenesis (including bone mineral density, relative bone volume, trabecular number and trabecular thickness) in group A were more evident than those in groups B and C (P<0.05). No significant differences were found between groups B and C (P>0.05).

CONCLUSIONLocal administration of recombinant adenovirus Ad-hAPN-EGFP may be an effective strategy to improve the restoration of bone defects in vivo.

Adenoviridae ; Adiponectin ; Animals ; Bone Density ; Bone and Bones ; Humans ; Osteogenesis ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; X-Ray Microtomography

Objective To clarify the MUM1/IRF4 expression in follicular lymphoma (FL) and its clinical and pathological significance. Methods Ninety-six cases FL were immunostained with MUM1,CD10,bcl-2,bcl-6 and Ki-67 antibodies. The results were compared with their clinical and pathological features. Results The overall MUM1 expression rate in FL was 59.2 % (58/96),including 36.2 % (19/51) grade 1 or 2 and 86.4 %(39/45) grade 3 cases (x2 =24.406,P <0.001). 68.9 % cases with diffuse area were MUM1 positive (x2 =8.161,P =0.004). MUM 1 and CD10 expression had inverse correlation,83.3 % CD10 negative cases were MUM1 positive (x2= 12.649,P<0.001). The mitosis rate and Ki-67 label index were statistically higher in MUM1 positive cases than in negative cases (t = -3.852 & -4.610,respectively,P <0.001). Conclusion MUM1 can be used as a biomarker to divide FL into different malignancies. The MUM1 positive FL may be the feature of high grade non germinal center B cell malignant lymphoma.

OBJECTIVETo construct a eukaryotic expression vector for expressing hepatitis C virus (HCV) recombinant NS5B-EGFP fusion protein and obtain a stable transfected HepG2 cell line.

METHODSThe coding region of NS5B gene of HCV was amplified by PCR and was digested by Xho I/Kpn I. This fragment was inserted into pEGFPN3 with T4 ligase and transformed E. coli TG1. The positive recombinant plasmid was selected, then the recombinant plasmid was transfected into HepG2 cell by Lipofectin AMINE 2000. Cells containing stable transformants were selected by the ability of resistance to G418 and isolated with a limited dilution. The stable transfected cell line expressing high level NS5B-EGFP fusion protein was obtained.

RESULTSThe eukaryotic expression vector named pEGFPN3-ns5b was successfully constructed and the stable transfected HepG2 cell line expressing NS5B-EGFP fusion protein was obtained.

CONCLUSIONThe stable transfected HepG2 cell line could express NS5B-EGFP fusion protein, could be used for anti-HCV infection with ns5b gene as the target.

Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Fluorescence ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Viral Nonstructural Proteins ; genetics ; metabolism

Animals ; Dose-Response Relationship, Drug ; Guinea Pigs ; Heart Ventricles ; Ion Channels ; antagonists & inhibitors ; physiology ; Male ; Myocytes, Cardiac ; drug effects ; physiology ; Promethazine ; pharmacology ; Terfenadine ; pharmacology ; Time Factors

OBJECTIVETo identify and characterize the epitope associated with the virus attachment protein (VAP) of hantaan virus.

METHODSThe monoclonal antibody 3G1 was used as the ligand to biospan from a phage-displayed 12-amino acid peptide library, then the positive phage clones were chosen and sequenced. The amino acid sequences of them were compared with that of hantaan virus G2 in homology. The characteristics of positive phage were studied by IFA and ELISA. A decapeptide combining to cell membrane was observed under laser scanning confocal microscope (LSCM).

RESULTSThe conservative motif PX(1-2) HX(0-2) H displaying on positive clones shared homologous amino acid sequence with G2 96YPWHTAKCHY105.

CONCLUSIONG2 96YPWHTAKCHY105 might play some roles in virus binding to host cell, and might be a possible key epitope of hantaan virus VAP.

Animals ; CHO Cells ; Cell Membrane ; metabolism ; Cercopithecus aethiops ; Cricetinae ; Cricetulus ; Enzyme-Linked Immunosorbent Assay ; Epitopes ; immunology ; metabolism ; Hantaan virus ; immunology ; Microscopy, Confocal ; Oligopeptides ; immunology ; metabolism ; Peptide Library ; Vero Cells ; Viral Proteins ; immunology ; metabolism

This study was purposed to establish the method of quantifying RhD antigen on red blood cells (RBC) by flow cytometry (FCM) and to explore the expression of D antigen on RBC of different RhD serotype. RhD(+) RBCs and RhD(-) RBCs were mixed in 1:1 ratio. Cells were stained by the indirect method (IgG anti-D as the first antibody, FITC-anti-IgG F(ab')2 as the second antibody), and the ratio of RhD(+) on RBCs was quantified by FCM. The optimal dosage of IgG anti-D was defined. Expression of RhD antigen on RBC of RhD(+), weak D, RhDel and RhD(-) type were detected by FCM. The results showed that optimal dilution of IgG anti-D monoclonal antibody was 1:4, 1x10(6) cells/50 microl. The percentage of D(+) RBC of RhD(+), weak D, RhDel and RhD(-) type were 96.8+/-2.97%, 79.5+/-9.88%, 47.8+/-11.43%, 3.7+/-2.96%, respectively. The mean fluorescence intensity (MFI) of RhD antigen expression of RhD(+), weak D, RhDel and RhD(-) type were 33.3+/-6.21 Dal, 18.6+/-5.39 Dal, 7.10+/-1.17 Dal, 0.79+/-0.55 Dal, respectively. In conclusion, there are significant differences of RhD antigen expressions among RBC of different RhD serotypes. The level of antigen on RhD(+) RBC is the highest and then weak D the next, while the level of antigen on RhDel RBC is the lowest level.
Blood Donors ; Erythrocytes ; immunology ; metabolism ; Flow Cytometry ; methods ; Humans ; Rh-Hr Blood-Group System ; immunology ; metabolism

BACKGROUNDY-27632 is a specific inhibitor of Rho-associated coiled kinase (ROCK) and has been shown to promote the survival and induce the differentiation of a variety of cells types. However, the effects of Y-27632 on adult human adipose tissue-derived stem cells (ADSCs) are unclear. This study aimed to investigate the effects of Y-27632 on the neuronal-like differentiation of ADSCs.

METHODSADSCs were isolated from women undergoing plastic surgery and cultured. ADSCs were treated with different doses of Y-27632 and observed morphological changes under microscope. The expression of nestin, neuron specific enolase (NSE) and microtubule-associated protein-2 (MAP-2) in ADSCs treated with Y-27632 was detected by immunocytochemistry and Western blotting analysis.

RESULTSY-27632 had the potency to induce neuronal-like differentiation in ADSCs in a dose-dependent manner. Moreover, the differentiation induced by Y-27632 was recovered upon drug withdraw. ADSCs treated with Y-27632 expressed neuronal markers such as NSE, MAP-2 and nestin while untreated ADSCs did not express these markers.

CONCLUSIONSelective ROCK inhibitor Y-27632 could potentiate the neuronal-like differentiation of ADSCs, suggesting that Y-27632 could be utilized to induce the differentiation of ADSCs to neurons and facilitate the clinical application of ADSCs in tissue engineering.

Adipose Tissue ; cytology ; Adult ; Amides ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Female ; Humans ; Neurons ; cytology ; Pyridines ; pharmacology ; Stem Cells ; cytology ; drug effects

Objective :To explore therapeutic effect and safety of irbesartan hydrochlorothiazide (I+H) tablet combined am-lodipine besylate on young and middle-aged patients with hypertension .Methods : A total of 138 young and middle-aged pa-tients with hypertension ,who were treated in our hospital from Jan 2015 to Feb 2017 ,were selected .Patients were ran-domly and equally divided into group A (received I+ H therapy) ,group B (received amlodipine treatment ) and group C (received I+ H+ amlodipine treatment ) ,all groups were continuously treated for three months .Systolic blood pressure (SBP) ,diastolic blood pressure (DBP) and heart rate (HR) before and after treatment ,total effective rate and incidence rate of adverse reactions were measured and compared among three groups .Results :Compared with before treatment ,after treatment ,there were significant reductions in SBP ,DBP and HR in three groups , P＜0.05 or ＜0.01. Compared with group A after treatment ,there was significant reduction in SBP [ (133.1 ± 6.2) mmHgvs.(128.9 ± 6.0) mmHg vs. (122.0 ± 3.8) mmHg] in group B and C ,and that of group C was significantly lower than that of group B ,P＜0.01 all ;compared with group B after treatment ,there was significant reduction in DBP [ (91.7 ± 6.7) mmHg vs.(87.5 ± 4.8) mmHg vs.(79.3 ± 3.0) mmHg] in group A and C ,and that of group C was significantly lower than that of group A ,P＜0.01 all.Compared with group A and B ,there was significant rise in total effective rate (78.3% vs.73.9% vs.97.8%) in group C , P＜0.01 all.There was no significant difference in incidence rate of adverse reactions among three groups , P＝0.876 .Conclusion : Irbesartan hydrochlorothiazide tablet combined amlodipine besylate tablet can significantly improve blood pressure in young and middle-aged patients with hypertension , Its effect is better than that of two drugs alone use . Which is safe and reliable ,and is worth extending .

OBJECTIVE: To observe the changes of relative expression of myocardial various RNAs in rats died of different causes and their relationship with PMI. METHODS: The rat models were established in which the rats were sacrificed by broken neck, asphyxia, and hemorrhagic shock. Total RNAs were extracted from myocardium. The quantitative real time PCR was used to calculate threshold cycle values of RNAs including glyceraldehyde-3-phosphate dehydrogenase (GAPDH), beta-actin, inducible nitric oxide synthase (iNOS), hypoxia-inducible factor-1 (HIF-1), tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and U6 small nuclear RNA (U6 snRNA) and to study the changes of the relative expressions of various indexes with PMI. RESULTS: U6 snRNA with stable expression level could be used as appropriate internal control. In the early PMI, the relative expression of GAPDH, HIF-1, iNOS, TNF-alpha, and IL-6 more characteristically increased in groups of asphyxia and hemorrhagic shock than in group of broken neck, but the quantity of beta-actin decreased in all groups. In the late PMI, all the relative expressions significantly declined in correlation with the degradation of RNA. CONCLUSION The characteristic changes of each RNA expression can be used as references to estimate PMI in deaths by different causes.
Actins ; Animals ; Cause of Death ; Cytokines/metabolism* ; Disease Models, Animal ; Enzymes/metabolism* ; Glyceraldehyde-3-Phosphate Dehydrogenases ; Myocardium/metabolism* ; Nitric Oxide Synthase Type II ; RNA/metabolism* ; RNA, Small Nuclear ; Rats ; Shock, Hemorrhagic ; Tumor Necrosis Factor-alpha

OBJECTIVEEndomyocardial biopsies from 42 (35 males and 7 females, aged 43.3 years) heart transplant recipients due to end-stage heart failure between June 2004 and January 2006 in our institute were obtained for pathological studies.

METHODSSixteen patients underwent 1 endomyocardial biopsy (right ventricular septum) between 13 days to 5 months, 13 patients underwent second biopsy between 1.5 to 8 months and 10 patients underwent third biopsy between 3 to 8.5 months post transplantation. Specimen were stained by hematoxylin-eosin (HE) and Phosphotungstic Acid Hematoxylin (PTAH) and observed under light microscope and cardiac allograft rejection were evaluated according to the Revision of the 1990 working formulation for the standardization of nomenclature in the diagnosis of heart rejection in 2004.

RESULTSThe rejection grades were as follows: Grade 0 R in 31 biopsies; Grade 1 R (mild rejection 1990 grade 1A, 1B and 2.) in 30 biopsies; Grade 2 R (moderate rejection, 1990 grade 3A) in 3 biopsies; Grade 1 R cellular rejection companies with humoral rejection in 1 biopsy. Cellular rejection with Quilty effect was found in 2 biopsies. Ischemic myocardial injury presented in 4 biopsies. Quilty effect was observed in 1 biopsy. Cytomegalovirus or toxoplasmic myocarditis was not observed.

CONCLUSIONSEndomyocardial biopsy (EMB) is a valuable diagnostic procedure for rejection surveillance in heart allograft recipients. The observed low rejection incidence and mild rejection from specimens of our heart recipients were comparable to the results of developed countries.

Adolescent ; Adult ; Biopsy ; Endocardium ; pathology ; Female ; Heart Failure ; pathology ; surgery ; Heart Transplantation ; Humans ; Male ; Middle Aged ; Myocardium ; pathology ; Postoperative Period