Objective To observe the effects of the bone marrow mesenchymal stem cells (BMSCs) on the expression of neurotrophic factor protein gene in the retinal detachment (RD) rabbits.Methods 60 healthy rabbits were randomly divided into control group (group A),retinal detachment with PBS group (group B),retinal detachment with BMSCs group (group C),20 rabbits in each group.RD model were established for rabbits in group B and C.10 μl PBS was injected into the subretinal space of rabbits in group B,while 10 μl CM-Dil labeled BMSC PBS was injected into subretinal space of rabbits in group C.The rabbits in the group A received no treatment.At 1,2 and 4 weeks after modeling,the mRNA expression of basic fibroblast growth factor (bFGF),brain derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) were measured by real-time quantitative PCR.Results At 1,2 and 4 weeks after modeling,the mRNA expression of bFGF,BDNF,CNTF on retinal tissue were increased significantly in group C as compared with group A and B (P＜0.01).At 1 week after modeling,the mRNA expression of bFGF and CNTF on retinal tissue were increased significantly in group B as compared with group A,the mRNA expression of BDNF on retinal tissue in group B was similar with group C.At 2 and 4 weeks after modeling,the mRNA expression of bFGF,BDNF,CNTF were decreased in group B as compared with group A.Conclusion Subretinal transplantation of BMSC can increase the mRNA expression of bFGF,BDNF and CNTF on retinal tissue in RD rabbits.

Humans ; Subacute Sclerosing Panencephalitis

OBJECTIVETo explore the effect of Danshensu on the lipid metabolism of hyperlipidemic rats.

METHODSixty clean male SD rats were selected. Twelve of them were selected in the basic control group and fed with common foods, and the remaining rats were fed with the high-fat feeds. After the successful modeling, they were randomly divided into the high-fat control group and low dose (10 mg x kg(-1) x d(-1)), medium dose (20 mg x kg(-1) x d(-1)) and high dose (40 mg x kg(-1) x d(-1)) Danshensu (dissolved in saline) groups. Both of the two groups were abdominally injected with the same volume of normal saline once a day for consecutively 30 days. The serum TG, TC, HDL-C and liver ACC1, FAS, HMGR, CPT-I mRNA expressions were detected.

RESULT AND CONCLUSIONDanshensu could inhibit the LDL-C level, timely clear redundant cholesterol and effectively regulate the lipid metablism of hyperlipidemic rats by reducing the TC content, decrease the fatty acid by reducing the FAS mRNA expression, and reduce the synthesis levels of endogenous cholesterol by inhibit the HMGR mRNA expression.

Animals ; Hyperlipidemias ; drug therapy ; metabolism ; Lactates ; pharmacology ; Lipid Metabolism ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley

Objective To investigate the short-cycle repeated transurethral resection ( Re-TURBT) combined pirarubicin treatment of T2 muscle invasive bladder tumor efficacy and safety.Methods From October 2010 to December 2013, 56 clinically diagnosed cases of bladder cancer were divided into two groups;the experimental group was taken a short-cycle repeat transurethral resection surgery, and the con-trol group taken along open surgery to remove the tumor and its surrounding 2cm bladder tissue, postopera-tive patients were given pirarubicin infusion therapy.Results Each patient completed the trial.For the ex-perimental group, blood loss and operative time was significantly lower than the control group ( P <0.05). For experimental group after 3 months, 6 months, and 12 months, maximum bladder capacity ( MBC) and patient perception of bladder condition ( PPBC) were significantly higher than control group ( P <0.05 ) . For experimental group after 3 months and 6 months, maximum urinary flow rate ( Qmax) was significantly higher than control group ( P <0.05) , but no significant difference between two groups at 12 months.The incidence of adverse events, vascular endothelial growth factor ( VEGF) , and a total recurrence rate showed no significant difference ( P >0.05).For experimental group, bladder perforation in 1 case, 1 case was forced to open surgery, the incidence rate was about 6.7%.Conclusions The short-cycle repeated trans-urethral resection combined pirarubicin treatment of T2 invasive bladder tumor was simple, effective and se-curity.

Objective To evaluate the efficacy and safety of 308-nm excimer light in the treatment of stable vitiligo. Methods Thirty patients with stable vitiligo were enrolled in this clinical trial. All the subjects received the treatment with 308-nm excimer light on a twice-weekly schedule for 3 months. Results The repigmentation rate was 95.0%, 75.0% and 66.7% for lesions in the face and neck, trunk and limbs, with the treatment sessions averaging 10.22 ± 1.60, 19.10 ± 2.38, 37.74 ± 3.06, respectively, and accumulative irradiation dose averaging 7.50 ± 3.45, 10.60 ± 1.01, 18.56 ± 3.05 J/cm2 respectively. Significant differences were observed in the repigmentation rate and treatment sessions between the lesions in the face and neck, trunk and limbs (all P ＜ 0.05). No severe side effects were seen during the treatment. Conclusion 308-nm excimer light is effective and safe for the treatment of vitiligo.

Objective To perform a preclinical test of a delineation software based on atlas-based auto-segmentation (ABAS),to evaluate its accuracy in the delineation of organs at risk (OARs) in radiotherapy planning for nasopharyngeal carcinoma (NPC),and to provide a basis for its clinical application.Methods Using OARs manually contoured by physicians on planning-CT images of 22 patients with NPC as the standard,the automatic delineation using two different algorithms (general and head/neck) of the ABAS software were applied to the following tests:(1) to evaluate the restoration of the atlas by the software,automatic delineation was performed on copied images from each patient using the contours of OARs manually delineated on the original images as atlases;(2) to evaluate the accuracy of automatic delineation on images from various patients using a single atlas,the contours manually delineated on images from one patients were used as atlases for automatic delineation of OARs on images from other patients.Dice similarity coefficient (DSC),volume difference (Vdiff),correlation between the DSC and the volume of OARs,and efficiency difference between manual delineation and automatic delineation plus manual modification were used as indices for evaluation.Wilcoxon signed rank test and Spearman correlation analysis were used.Results The head/neck algorithm had superior restoration of the atlas over the general algorithm.The DSC was positively correlated with the volume of OARs and was higher than 0.8 for OARs larger than 1 cc in volume in the restoration test.For automatic delineation with the head/neck algorithm using a single atlas,the mean DSC and Vdiff were 0.81-0.90 and 2.73％-16.02％,respectively,for the brain stem,temporal lobes,parotids,and mandible,while the mean DSC was 0.45-0.49 for the temporomandibular joint and optic chiasm.Compared with manual delineation,automatic delineation plus manual modification saved 68％ of the time.Conclusions A preclinical test is able to determine the accuracy and conditions of the ABAS software in specific clinical application.The tested software can help to improve the efficiency of OAR delineation in radiotherapy planning for NPC.However,it is not suitable for delineation of OAR with a relatively small volume.

HBV polymerase has intrinsic RNA-dependent reverse transcriptase, DNA-dependent DNA polymerase as well as RNaseH activity. Analysis of HBV polymerase has been hampered for many years due to the inability to express functional enzyme in a recombinant system. To obtain active polymerase at a high level, we have taken advantage of baculovirus expression system. The gene of HBV polymerase was amplified by PCR and cloned into pFastBac Dual to construct the recombinant plasmid pFastbac Dual-pol. The recombinant donor plasmid, pFastbac Dual-pol, was constructed by inserting HBV polymerase gene into EcoRI and PstI sites controlled by polyhedrin promoter. The recombinant donor plasmid was transformed into DH10Bac competent cells for transposition. Recombinant bacmid was constructed by inserting of the mini-Tn7 element from the donor plasmid into the mini-attTn7 attachment site on the bacmid. The recombinant bacmid DNA was isolated and transfected into the Sf9 cells to produce the recombinant virus, and healthy insect Sf9 cells were infected with the recombinant virus containing HBV polymerase gene to express the target protein. HBV polymerase expressed in insect cells was analyzed by SDS-PAGE. PCR results showed recombinant donor plasmid, pFastbac Dual-pol, was constructed successfully. The recombinant hepatitis B virus polymerase was expressed in insect cells at high level. The recombinant hepatitis B virus polymerase should facilitate the analysis of HBV polymerase biological characteristics, allow the investigation for new anti-HBV drugs specifically blocking HBV polymerase.

Objective To investigate the clinicopathological features,diagnosis,treatment,prognosis and causative agent of a case of eumycetoma on the submaxilla.Methods A case of eumycetoma diagnosed in our department was assessed for its clinical and pathological features as well as mycologic and molecular identification.Related literature was reviewed.Results The patient was primarily characterized by swelling of the submaxilla,with multiple sinuses draining many black granules.Pathologic examination revealed a pyogenic granulomatous inflammation,and a number of lotus rhizome node-like hypha were observed in tissue samples through PAS staining.Sequence analysis of multiple loci of the isolates,including ITS 1,ITS2 and D1/D2,showed that it was mostly similar to Madurella mycetomatis with a homology of 97%.Conclusion This is a case ofeumycetoma on the submaxilla induced by a novel species of Madurella.

Objective To analyze the genetic characteristics and the evolution of the influenza A (H3N2) virus strains circulating in Jiangsu province between 2013 and 2014.Methods This study analyzed thirty-one representative strains of influenza A (H3N2) virus, which were isolated in different regions of Jiangsu province and during different time periods from 2013 to 2014.Results Genetic distances in nucleic acid and amino acid between a strain used for vaccine production (A/Texas/50/2012) and the 31 strains were 0.010 5 and 0.012 4.Similarities between them in nucleic acid and amino acid sequences were 97.9%-99.6% and 97.2%-99.3%.Phylogenetic analysis showed that the hemagglutinin (HA) genes of the 31 strains were divided into three different groups.Three strains isolated in 2013 and three strains isolated in 2014 belonged to Group 1 and Group 2, respectively, while the others belonged to Group 3.Three positive selection sites (237, 366 and 367) in HA protein were observed by REL model.Compared with the strain used for vaccine production, the 31 strains were characterized by amino acid substitutions (N128A/T and P198S/A) in HA protein and all of the mutations located in B-cell epitopes.The total number of mutation sites reached 24.Compared with the A/Texas/50/2012 strain, seven strains presented the glycosylation site 126NWT, and three strains showed disappeared glycosylation sites of 45NSS and 144NNS.Evaluation of vaccine efficacy for A(H3N2) virus strains showed that the vaccine efficacy was not very well.Conclusion The HA gene of A(H3N2) virus had undergone a greater variation and the vaccine efficacy was not very well in Jiangsu province during 2013 to 2014, which made the influenza A(H3N2) virus become the circulating strain.

It is often necessary to construct more than one recombinant plasmids when investigating the characteristics, physchemical features and functional mechanisms of genes or proteins. Repeated sub-cloning procedures including design of primers, enzyme digestion, ligation and verification of recombinant plasmids, have to be involved with. For this reason, it has become a tendency to developing new genetic vectors which can be used in multitude of experiments. Therefore, by using pIRES vector as a backbone, here we reported the construction of a mammalian expression vector: pCMV-Myc-IRES-EGFP which contains the N-terminal c-Myc epitope tag and the enhanced green fluorescent protein (EGFP) translated in an IRES-dependent manner. This novel vector can be used to testify the efficiency of cell transfection, to collect successfully transfected cell population via cytometry, to conduct transcription and translation in vitro, to purify target proteins or to trap their interactional proteins. The availability of this vector can facilitate function study of genes.
Apoptosis Regulatory Proteins ; genetics ; metabolism ; Base Sequence ; Blotting, Western ; Cell Line ; Cloning, Molecular ; Gene Expression ; Genes, myc ; genetics ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Fluorescence ; Molecular Sequence Data ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection