Common review comments on investigator initiated study (IIS) design and considerations / 中华医学科研管理杂志

Objective More and more investigator initiated studies have been funded in China.Both institutions and investigators should take the responsibility for meeting the scientific validity,ethical requirement,feasibility and interdisciplinary management requirements during the clinial research project design and initiation.This article hereby analyzed and summarized the most common review comments on clinical research applications.Methods Summarize the problems identified during the review of Shenzhen Second People's Hospital Clinical Research Program applications.Results The most common findings in turn as follows:inappropriate sample size calculation,study design,parameters,unclear study aims,insufficient study rational.Conclusions To fund clinical research programs with significant scientific values and appropriate design,it is vital for the research management department to provide more supervision and technology support.

Preparation and characterization of monoclonal antibody against hTSC29 pep-tides / 中国免疫学杂志

Objective:To prepare monoclonal antibody ( mAb ) against human testis-specific conserved gene ( hTSC29 ) peptides and characterize its immunological and biological features.Methods:According to bioinformatics analysis and prediction of the antigenicity, surface property, hydrophilicity and flexibility of hTSC29, a 18-amino acid residue partial peptide of hTSC29 was synthesized,then immunized the BALB/c mice for preparing antiserum.The mAb against hTSC29 was produced using the routine hybridoma technique.The properties of the mAb against hTSC29 were identified by ELISA, Western blot and immunohistochemistry staining.Results:After cell fusion and subcloning, one hybridoma cell lines secreting specific mAb against hTSC29 protein were obtaind.The Ig subclass of the mAb was IgG2b(κ).ELISA detection showed that the titer of mAbs in cultured was 1∶104.Western blot analysis proved that the mAb could specifically recognize Mr 60 000 protein in human testis total protein.The hTSC29 protein main located at circumference of spermatocyte and spermatid in human testis tissue by immunohistochemistry staining and immunofluorescence assay.Conclusion:One hybridoma cell lines which can secrete specific mAb against hTSC29 protein with high titers and specificity have been established successfully.The mAb will provide efficient tools for functional studies of hTSC29 expressed in spermatogenesis.

Expression of Tpap gene in mouse testis / 中国组织工程研究

BACKGROUND: These serial processes for forming male gametes are basically controlled by the programmed expression of a number of stage-specific genes. However, many aspects of the mechanisms of spermatogenesis have remained elusive because of a lack of suitable in vitro or in vivo models.OBJECTIVE: To screen genes involved in spermatogenesis, and to analyze its expression characteristics. METHODS: Testes cDNA samples from Balb/C mice of different postnatal days (4,9,18,35, 54 days and 6 months, respectively) were hybridized with mouse whole genome Affymetrix chip to screen the testis-ralated genes. The characteristics of the selected genes were analyzed by various bioinformatics tools. RT-PCR was used here to identify the expression of the selected genes in mice testis.RESULTS AND CONCLUSION: The Affymetrix chip probe of mouse Tpap was graduated higher expression with developmental stages of mouse testis. The scaling hybridization signal intensities of the tested testis on days 4, 9,18, 35, 54, and 6 months of postnatal were 4.4 (Absent expression, A), 12.9 (A), 262.4 (Present expression, P), 1136.7 (P), 1617.5 (P) and 1128 (P),respectively. These results indicated that the expression of mouse Tpap wasn't detected on days 4 and 9, but was detected on days 18, 35, 54, and 6 months of mouse testis in our Affymetrix chip analysis. By combination with the RT-PCR analysis of mouse Tpap, we observed mouse Tpap began to express at the age of day 18 in mouse. Tpap is an age-dependent gene in mouse testis.The expression of Tpap corresponds to the appearance of spermatids of mice and indicates that Tpap may have an important role in male mammalian spermatogenesis.

Expression profile of a novel germ cell-specific gene, TSCPA, in mice and human / 华中科技大学学报（医学）（英德文版）

In order to identify novel genes involved in spermatogenesis, testis cDNA samples from Balb/C mice of different postnatal days were hybridized with the whole mouse genome Affymetrix chip to screen the testis-specific genes. The characteristics of the selected genes were analyzed by RT-PCR as well as other bioinformatic tools. A novel differentially expressed testis-specific gene (GenBank Accession No: NM_029042) in the developmental stages of testes was identified, and named TSCPA. Cellular mapping prediction of TSCPA indicated that its protein was probably expressed in nuclei, and one putative domain (aa 332-377) was anchoring domain of cAMP-dependent type II PK. The result of subcellular localization of GFP-TSCPA fusion protein in Cos-7 cells showed that TSCPA protein was expressed in nuclei. RT-PCR analysis revealed that TSCPA was expressed specifically in mouse and human testis. TSCPA gene was expressed weakly in 21-day-old mouse testis and the expression was increased gradually from 38th day to 6th month of mouse testes. No expression of hTSCPA was found in cryptorchidism and Sertoli-cell-only syndrome patients. It was concluded that the expression profile of TSCPA in human and mice indicated that TSCPA might play an important role in spermatogenesis.