Human gene therapy needs to express exogenous DNA at the targeting cells,producing a practical and efficient therapeutic dosage at an approp-riate time(quantitative pharmacology)with a safe man-ner.Recombinant adeno-associated virus(rAAV)Vec-tom possess a number of properties and recent progress in rAAV production made it rapidly become the reagent of choice for therapeutic gene tmasfer.Over 60 clinical trials of gene therapy based on rAAV have been carried out.The dose response reaction between rAAV vectors and gene expression activity or clinical outcome is one of major aspects of these trials.Most studies showed that vector genomes(vg)and gene expression had a concentration-dependent relationship during a certain scope.However,gene expr~sion Can be afffected by viral serotypes,tissue tropisms,cell targeting,drug regulation,injection route,age and sex,etc.Thus,these aspects should be carefully comidered by scienti-sts,pharmacologisis and physicians during animal ex-periments or clinical trails.KEY WORDS gene therapy;viral vector;dose-re-sponse;quantitative pharmacology;clinical thempy

Pancreastatin, a novel peptide isolated from porcine pancreas, is capable of inhibiting the insulin release and the exocrine secretion of the pancreas. In this study, the localization and distribution of pancreastatin in guinea pig, pig and human pancreas were investigated with ABC immunostaining method combined with the glucose oxidase-DAB-nickle developing technique on paraffin sections. The results showed that in human pancreas pancreastatin-like immunoreactive(PLI)cells were mainly dist- ributed in the periphery of the islets. While in guinea pig and pig pancreas, the majority of islets cells showed positive pancreastatin immunostaining. The coexistence of pancreastatin and insulin in the pancreatic islets of guinea pig and pig were also proved with immunostaining of adjacent thin sections. PLI-cells were also found in the epithelium of the ducts and acini of all investigated pancreas. However, in the exocrine portion of human and pig pancreas, PLI-cells were present in amounts smaller than those found in guinea pig pancreas. The significance of the distribution pattern of pancreastatin was discussed.

Chinese medical student's education system is quite different from that in western countries.Besides,Chinese residency training program is just starting.The professional training of pathologists,who make the final diagnosis in the hospital,is not satisfied.The present situation in China is that overall quality of pathologists is low and varied.The pathology department of Xijing Hospital started to carry out the pathology residency training program since 2009.It is expected to search for a model of pathology residency training program with Chinese characteristics.

Objective:To construct K-ras-targeted siRNAs (K-ras siRNA) and to investigate the inhibitory effects of K-ras siRNAs on the growth and migration of lung cancer A549 cells (containing mutant K-ras gene) and NCI-H446 cells (containing wild-type K-ras gene). Methods: Four K-ras siRNAs (K-ras siRNA1～K-ras siRNA3 targeting wild-type K-ras and K-ras siRNA4 targeting mutant K-ras) were designed and artificially synthesized; they were used to transfect A549 cells and NCI-H446 cells. The expressions of Ras mRNA and protein were examined by RT-PCR and Western blot-ting. The inhibitory effects of K-ras siRNAs on the proliferations of A549 and NCI-H446 cells were determined by MTT assay. The effects of K-ras siRNAs on the cell migration and apoptosis were observed by Transwell assay and Hoechst 33258 staining, respectively. Results: Mutant K-ras-targeted siRNA (K-ras siRNA4) specifically inhibited the K-ras ex-pression but had no influence on H-ras and N-ras expression in A549 cells. K-ras siRNA4 inhibited the proliferation of A549 cells but did not inhibit that of NCI-H446 cells, which contained wild type K-ras gene. K-ras siRNA4 also induced apoptosis and inhibited migration of A549 cells. Conclusion: Mutant K-ras-targeted siRNA4 can inhibit the proliferation, migration and induce apoptosis of A549 cells. It may be a potential and personalized drug for the treatment against lung cancer containing mutant K-ras gene.

Gene medicine based on recombinant adeno-associated virus (rAAV) vector has rapidly become the prior-choose reagent for gene therapy, since it had been shown that the rAAV was able to stably express many genes in vivo without detectable side-effect. However, recent findings of CTL immune responses to AAV capsid in a clinical trial highlighted a new issue regarding safety that previously was not identified in animal studies. Obviously it is so important to understand the interaction of rAAV with the immune system in details for the safety and success of rAAV gene medicine. In this review we evaluate several current hypotheses aiming to explain the cellular immunotoxicity, also analysis the current findings including the presentation kinetics of the capsid antigen and the activation of CTL. Focusing on the key steps of the immune response several solutions are proposed, including immunosuppression, optimization of vector and improvement of purity, in order to insure clinical safety and efficacy of rAAV.

This paper introduces the design and implementation of a system which can get the NEMA2.0 image data from the hard disks of the imaging equipments directly,then analyzes and transforms these image data into the DICOM3.0 image data and sends them to the image server. The design has the advantages of reliable image quality, low cost and information.
Computer Storage Devices ; Diagnostic Imaging ; Image Processing, Computer-Assisted ; methods ; Signal Processing, Computer-Assisted ; Software

Purpose To investigate human soluble TRAIL(sTRAIL)protein expression and purification and its potential anti-tumor activity on hepatocellular carcinoma(HepG2).Methods Soluble TRAIL gene ligated with expression vector pPIC9 was transfected into GS115(his4)and the recombination strain expressing sTRAIL was screened by MD plate.The effects of different media,methanol inducement period,methanol concentration,and pH were investigated and optimized using shaking flask.The anti-tumor activity of sTRAIL with HepG2 cells Was analyzed after purification.Results The highest expression of sTRAIL was obtained at pH 6.0,1% methanol in BMMY medium,with the concentration of(58.7±2.4)mg/L at 48 h.Recombinant sTRAIL protein could induce HepG2 cells apoptosis and inhibit HepG2 cells proliferation effectively.Conclusion The optimized condition of human sTRAIL expression and purification Was developed and the obtained recombinant sTRAIL protein may be a promising therapeutic agent for hepatocellular carcinoma.

Objective To investigate the effects of infrasound at different frequencies on the pathological morphology of mouse testes and to explore the possible involvement of β-cateinin in the mechanism of injury. MethodsSixty adult BALB/C mice were randomly divided into 2 groups.One group of 6 mice were the controls without infrasound exposure.The other group with 54 mice was evenly divided into 3 subgroups of 18 exposed to 130 dB infrasound at 4 Hz,8 Hz or 20 Hz for 2 hours daily.The pathological morphology of the mice's testes was observed 3,9 and 21 h after exposure.Immunohistochemical staining was used to examine β-catenin expression 3 h after exposure.ResultsMarked damage was observed in the morphology of the testicular cells of rats in the 8 Hz group.Cell denaturation was found in germinal cells.The cell structure was loosened and the cell junctions had decreased or disappeared.The damage was most serious at 3 h after exposure.But at the same time point,pathological damage of testicular cells was not obvious in the 4 Hz group,with only a few testes cells swollen.The morphology of testicular tissue of mice in the 20 Hz subgroup exhibited no significant difference from the control group.β-catenin was mainly found in the membranes of Leydig cells,Sertoli cells and myoepithelial cells in all of the mice,and there was no significant change in the expression level or distribution pattern between the exposed mice and the controls.No expression ofβ-catenin was found in spermatogenic cells. ConclusionInfrasound at 4 Hz or 8 Hz can damage testicular tissue,at least in mice.At 20 Hz there is no significant effect on the testes.The loosened cell structure might have been caused by the mechanical force produced by the infrasound and not by any decrease of β-catenin.

High mobility group A2 protein (HMGA2), an architectural factor, is highly expressed in various cancer types including lung cancers. It is a candidate target for cancer therapy. RNAi is an effective gene silencing method with low cost and less time-consuming. It is possible to exploit this technology in therapy. Here, 5 siRNAs targeting Hmga2 gene (HMGA2 siRNA1-5) were designed and synthesized. MTT assay, colony formation assay, transwell assay and flow cytometry were used to evaluate the effects of these siRNAs on lung cancer cell lines (NCI-H446 and A549). Results from cell proliferation, clone formation, migration and apoptosis showed that HMGA2 siRNA1, 3, 5 could affect these aspects for both lung cancer cell lines. Among the five siRNAs, HMGA2 siRNA5 showed the greatest inhibition effects. The inhibition effects of HMGA2 siRNA5 are sequence specific and are not due to the induction of interferon response. Taken together, siRNAs targeting Hmga2 gene are potential candidates for lung cancer gene therapy.

A safe and effective targeting viral vector is the key factor for successful clinical gene therapy. microRNA, a class of small, single-stranded endogenous RNAs, act as post-transcriptional regulators of gene expression. The discovery of these kind regulatory elements provides a new approach to regulate gene expression more accurately. In this review, we elucidated the principle of microRNA in regulation of targeting viral vector. The applications of microRNA in the fields of elimination contamination from replication competent virus, reduction of transgene-specific immunity, promotion of cancer-targeted gene therapy and development of live attenuated vaccines were also discussed.
Gene Expression Regulation ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; MicroRNAs ; genetics ; Viruses ; genetics ; metabolism