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.

The report expounds the importancr of 'jiangxi treatment' by discussing the meaning,the researches in the past dynasties and the collation of variety methods of 'jiangxi treatment'. It thinks that the 'jiangxi treatment'in TCM includes the right way to decoct drugs,the rational way to take drugs and the variety ways to 'jiangxi 'after taking drugs. Thus,it should be used selectively in clinical. And the report suggests that the 'jiangxi treatment' is a effective way to increase the clinical eff icacy of drugs.

Objective To explore the evolution and treatment of traumatic subdural effusion(TSE).Methods The clinicsl materials of 66 patients with TSE were analyzed retrospectively.Results 53 patients were cured with comervative therapy,and other patients were evolved into chronic subdural hematoma(CSDH).8 patients with CSDH were cured with surgery and others with conservative therapy.Conclusion Patients with TSE don't need surgery,and then patients with clinical characteristics will be operated when TSE evolves into CSDH.

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.

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.
Animals ; Cell Differentiation ; Cells, Cultured ; Embryonic Stem Cells ; cytology ; Insulin-Secreting Cells ; cytology ; Mice ; Neural Stem Cells ; cytology

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.

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.

Objective Although principal components analysis profiles greatly facilitate the visualization and interpretation of the multivariate data,the quantitative concepts in both scores plot and loading plot are rather obscure.This article introduced three profiles that assisted the better understanding of metabolomic data.Methods The discriminatory profile,heat map,and statistic profile were developed to visualize the multivariate data obtained from high-throughput GC-TOF-MS analysis.Results The discriminatory profile and heat map obviously showed the discriminatory metabolites between the two groups,while the statistic profile showed the potential markers of statistic significance.Conclusion The three types of profiles greatly facilitate our understanding of the metabolomic data and the identification of the potential markers.

Objective To investigate the mid-term clinical outcomes of arthroscopic anatomical re-construction of double bundles of the anterior cruciate ligament ( ACL ) . Methods The clinical data of 78 patients diagnosed with ACL rupture from April 2012 to July 2014 were analyzed retrospectively. They were 60 males and 18 females, aged from 19 to 56 years ( mean, 26. 8 years ) . The time from injury to surgery ranged from one week to 23 months ( mean, 5. 8 months ) . All of them obtained positive results in anterior drawer test and Lachman test preoperatively. Their preoperative KT-1000 examinations showed an average side-to-side difference of 8. 29 ± 1. 81 mm in anterior laxity. They were all treated with arthroscopic anatomical recon-struction of double bundles of the anterior cruciate ligament using autologous hamstrings. The International Knee Documentation Committee ( IKDC ) and Lysholm scores were used to assess their knee function at the last follow-up. Results All the 78 patients were followed up for 34. 6 months on average ( range, from 25 to 56 months ) . At the last follow-up, the IKDC and Lysholm scores were significantly increased from preoper-ative 42. 6 ± 9. 5 and 44. 4 ± 8. 5 to postoperative 92. 9 ± 2. 8 and 94. 2 ± 3. 4, respectively ( P <0. 05 ) . The Lachman test was negative in 73 cases ( 93. 6％) and the pivot shift test was negative in 69 cases ( 88. 5％) . The KT-1000 examinations showed that the side-to-side difference in anterior laxity averaged 1. 47 ± 0. 68 mm, significantly improved from the preoperative values ( P <0. 05 ) . At their last follow-up, 29 patients underwent MRI scans which showed continuity of the anteromedial and posterolateral bundles of the anterior cruciate liga-ment. Conclusion The arthroscopic anatomical reconstruction of double bundles of the anterior cruciate ligament can restore the knee stability and achieve fine mid-term clinical outcomes.

Objective: To investigate the impact of minimal residual disease (MRD) by multiparameter flow cytometry (MPFC) during aplasia on efficacy and prognosis of de novo acute myeloid leukemia (AML) (non M₃) patients. Methods: The MRD data by 8-color MPFC during aplasia (day 14-15 of induction therapy) in 85 de novo AML (non M₃) patients and the MRD impact on efficacy and prognosis were retrospectively analyzed. Results: Data of 85 patients, including 42 males (49.4%) and 43 females (50.6%) , were collected, with a median age of 35 (15-54) years. The median MRD by MPFC during aplasia was 0.58% (0-81.11%) , and 70 (82.4%) patients achieved complete remission (CR) after first induction chemotherapy. The cutoff of MRD by receiver operating characteristic (ROC) analysis was 2.305% (Se= 0.867, Sp=0.800) . The CR rate after one course was significantly higher in patients with MRD<2.305% [96.6% (56/58) ]than in patients with MRD≥2.305%[51.9% (14/27) ] (χ²=22.348, P<0.001) ; no significant difference with respect to relapse-free survival rate (χ²=1.08, P=0.299) or overall survival rate (χ²=0.42, P=0.516) could be demonstrated for the comparison of the two groups. Multivariates analysis showed MRD divided by 2.305% was the only independent prognostic factor for CR after one course (OR= 21.560, 95% CI 4.129-112.579, P<0.001) . Conclusion Flow cytometric MRD divided by 2.305% during aplasia could be a predictor of efficacy after first induction therapy in AML patients.