Clinical application of semiconductor gene sequencing in screening thalassemia / 实用医学杂志

Objective To investigate the feasibility of semiconductor gene sequencing technology for thalassemia clinical screening and evaluate its application as compared with the results of PCR technology.Methods 197 visiting patients were randomly selected as prospective samples and200 patients ever diagnosed with thalassemia as previous samples.All the samples were detected by semiconductor technology gene sequencing and PCR technology at the same time and then evaluation of the advantage of semiconductor gene sequencing technology.Results 22 cases of 197 prospective samples were detected as thalassemia mutations by PCR technology,including 18 cases of α-thalassemia,3 cases of β-thalassemia,1 case of oα merge β thalassemia mutations.Semiconductor technology gene sequencing detected another 6 cases of rare type of thalassemia.By semiconductor gene sequencing technology on previous samples,118 cases of α-thalassemia,65 cases of β-thalassemia,17 case of α merge β thalassemia mutations,1 case of thalassemia mutations (HBA 1:c.223G ＞ C) were detected.By statistical analysis,the total coincidence rate of PCR technology and semiconductor gene sequencing was 98.5％,withthe Kappa =0.97(≥ 0.8).Conclusion Semiconductor gene sequencing technology for thalassemia clinical screening is feasible,for it can detect both thalassemia gene type,and new mutation.The results of semiconductor gene sequencing technology are accurate and the technology could be popularized in clinical application.

Induced pluripotent stem cell therapy：transplant rejection and safety / 中国组织工程研究

BACKGROUND:Induced pluripotent stem cels have been a hotspot in regenerative medicine research since it was discovered. The clinical application of induced pluripotent stem cels is excessively focused on, but the safety issue is almost ignored. OBJECTIVE:By summarizing the application of induced pluripotent stem cels in animal experiments to analyze the safety problems of induced pluripotent stem cels and their possible reasons in order to lay a foundation for further study and clinical application of induced pluripotent stem cels. METHODS: PubMed database was retrieved by the first author for articles related to the safety of induced pluripotent stem cels published from 2006 to 2014 using the keywords of “induced pluripotent stem cels, safety, immune, immunogenicity, tumorigenicity, cancer, epigenomic, transplantation, generation, reprogramming,genomic, mutation” in English. Related ful texts were got from Cel Press and Nature Databases. Finaly, 28 articles were chosen in result analysis. RESULTS AND CONCLUSION: Safety problems of induced pluripotent stem cels are attracting more and more attentions. Immunogenicity, potential tumorigenicity and epigenetic variation are major risks for the clinical applications of induced pluripotent stem cels. Safety issues of induced pluripotent stem cels mainly come from the reprogramming process. The “integrating genetic manipulation” may lead to a greater risk of tumorigenicity than non-integrating operations. Epigenetic variations emerge in the reprogramming, which are mostly relative to “epigenetic memory” of reprogrammed cels.

Study on the course reformation of medical laboratory science after changing five-year system to four-year system / 中华医学教育探索杂志

To adapt to the system reformation of medical laboratory science from five academic years to four academic years and to meet the new professional technology-oriented requirements, the medical laboratory science institute of Guangdong Medical College has carried out a comprehensive reform of curriculum system. This paper has analyzed the current problems in the school medical ex-amination and explored the curriculum system reform from three respects such as adjusting curriculum by restructuring and integrating programs, implementing modular teaching to build its characteristics and strengthening practice teaching.And it has also explored the full assessment mode by optimizing the traditional one-stop assessment.

Neural-precursor-cells-phase is not the indispensable checkpoint of embryonic stem cell differentiation into insulin-producing cells / 生物医学工程学杂志

Investigation on the pathways of embryonic stem cells differentiation into insulin-producing cells is of importance to pancreatic tissue-engineering. Instead of passing through the classic multi-step-inducing method, the expanded embryonic stem cells that were cultured and expanded in the presence of mouse embryonic fibroblast feed-layer and leukemia inhibitory factor (LIF) were induced into insulin-producing cells directly. The results showed a similar consequence from two different inducing cultures. Without passing through a so-called indispensable differentiation phase, the neural-precursor-cell-stage, the expanded embryonic stem cells could be induced into insulin-producing cells. The insulin-producing cells population resulting from our modified method were similar to that resulting from the classic multi-step method (passing through the neural-precursor-cells-stage), thus suggesting that neural-precursor-cell-phase is not the indispensable checkpoint of embryonic stem cell differentiation into insulin-producing cells. Embryonic stem cells can be induced into insulin-producing cell by classic multi-step inducing method or by direct inducing method.

Construction of mouse pdx-1 gene eukaryotic expression vector and its expression in embryonic stem cells / 中国病理生理杂志

AIM: To clone mouse pdx-1 gene and construct its eukaryotic expression vector for expression of pdx-1 in mouse embryonic stem cells.METHODS: Mouse pdx-1 cDNA fragment was amplified with polymerase chain reaction (PCR) from mouse pancreatic cDNA. The purified fragment was recombinated with a eukaryotic expression vector carrying enhanced green fluorescent protein, pEGFP-N1. The pdx-1 cDNA fragment was inserted into the multi-clone sites of the vector to construct a new plasmid, pEGFP/pdx-1. E.colli strain DH5? was transfected with the new recombinant plasmid to expand it. Plasmid DNA extracted from the expanded DH5? was identifed by cutting with Hind Ⅲ, BamHⅠ nuclease and by DNA sequencing. Identified plasmid DNA was transfected into mouse embryonic stem cell line MESPU13 by carrying with liposome. RESULTS: A 876 bp cDNA fragment was amplified from mouse pancreatic cDNA by PCR and it was inserted into the vector pEGFP-N1 correctly. The fragment was defined to be pdx-1 gene by nuclease digestion and DNA sequencing. Mouse embryonic stem cell line MESPU13 was transfected with the new recombinant plasmid DNA. The green fluorescent protein report gene and pdx-1 gene expressed in transfected mouse embryonic stem cells within 24 h. CONCLUSION: Mouse pdx-1 gene is cloned and its recombinant eukaryotic expression vector carrying green fluorescent protein is constructed successfully. It provides a useful tool for further research on the function of pdx-1.