Objective To evaluate the feasibility of transfection of Sonic hedgehog gene (SHH)into bone marrow mesenchymal stem cells(BMMSC).Methods After the SHH gene was transfected into BMMSC by electroporation apparatus,the transfection rate was evaluated by fluorescence inverted microscope.The growth curves of untransfected and transfected BMMSC were drawn,respectively,to observe the influence of transfection on cells.The expression of SHH gene in the BMMSC was detected by PCR,RT-PCR,Western-blot analyses.Results Through fluorescence inverted microscope,the observed transfection rate was appropriately 30％,PCR showed a obvious increase of SHH expression in transfected cells than that in untransfected cells,and it is quantified by qPCR for appropriately 7 times.Western-blot further demonstrated that the SHH protein expression in transfected cells had a distinct increase.However,it was observed that the exponential phase of BMMSCSHH growth curve delayed.The growth curves of both overlap 12 days after transfection.Conclusions This electroporation method can transfect exogenous SHH gene into BMMSC sufficiently with the effective protein expression in BMMSCSHH.It is the foundation of further research of genetic therapy for ischemic heart disease.

Teaching and research are two major functions of university. We should correct the misunderstanding of the relationship between teaching and research and improve the evaluation of integration of them. Positive interaction of research and teaching can enhance educational results of university. We should conduct the interaction of scientific research and teaching,improving the creative ability of students.

Objective To investigate the survival and differentiated situation of transplanted mesenchymal stem cells ( MSCs) transfected with Sonic hedgehog (SHH) gene after chronic myocardial infarction (MI).Methods MSCs were obtained from bone marrow of limb bones by density gradient centrifugation combined with attachment culture method .SHH gene was cloned and eukaryotic expression plasmid pcDNA 3.1-SHH was constructed .Then nucleofector TM was used to transfer SHH gene into MSCs and the expres-sions of mRNA and protein of SHH gene were detected in vitro .Myocardial infarction model was established by ligating the left anterior descending branch .A total of 150μl (3 ×106 ) of MSCs transferred with SHH gene was injected into ischemic area at the fourth week after MI model was set up .Brdu immunohistochemical staining and electron microscope examination were used to detect the survival and differentiation of MSCs with SHH in vivo at the 1st, 2nd, 4th, and 8th week after MSCs SHH transplantation.Results It was con-firmed that SHH gene clone and the recombinant eukaryotic expression vector pcDNA 3.1-SHH construction were successful by gene se-quence analysis and double digestions with EcoR I and Bamh I .Reverse transcriptase polymerase chain reaction ( RT-PCR) and West-ern-blot demonstrated that the expression of SHH mRNA was significantly increased in transfected MSCs compared with untransfected MSCs.Brdu immunohistochemical staining confirmed MSCs with SHH were survived from one to eight weeks in MI area after transplan -tation.Electron microscope examination showed that MSCs had the trend to be differentiated into myocardiac -like cells in MI area at the 4th and 8th week after transplantation .Conclusions The expression of SHH mRNA was significantly increased while MSCs were transfected with SHH gene , and MSCs SHH could survive and differentiate in good condition in the transplanted recipients .Those data provide the experimental basis for further transgenic cell therapy research of ischemic heart disease .

Objective To discuss the features,such as clinical symptoms,pathologic morphologies,immunohistochemical staining of minimal deviation adenocarcinoma and microglandular hyperplasia of the uterine cervix in order to improve the accuracy of pathological diagnosis.Methods s:Histopathologic characteristics of total hysterectomies in 2 cases of minimal deviation adenocarcinoma and 1 case of cervical microglandular hyperplasia based on the formalin-fixed,paralfin-embedded and hematoxylin-eosin stained tissue were analyzed retrospectively.Immunohistochemical staining was used to detect the expression of CEA,p53,PCNA,and Ki-67 in all 3 cases.Results The main clinical symptoms of minimal deviation adenocarcinoma were watery leucorrhea and enlargement of the cervix.The pathological findings of MDA included hyperplasia of the glands with cytological minimal atypia,invasion effects into the stroma could be observed in some glands and abortive glands with desmoplastic changes,or edema and inflammatory infiltration around the glands were also observed.The invasion presented in the deep part of the cervix as well.The patiant of MGH had a history of oval contraceptive use.Histological features of MGH included tightly packed glands in different sizes and shapes,presentation of inflammatory cells in stroma and glandular lumens,and focal epithelial cell pleomorphism and hyperchromatism but without mitosis.CEA was positive in all two MDA cases,but the tissue of MGH was negative for CEA.The expressions of the other four markers had no difference between MDA and MGH.Conclusions For patients with watery discharge and/or hypertrophy of cervix,the deep ( ＞ 5 mm ) biopsies should be performed.The immunohistochemical staining for CEA,p53,CA125 and ER has adjuvant diagnostic values.It is extremely important to recognize that MGH is an entirely benign lesion.

Objective:To study the regulatory effect of miR-9 on the proliferation of breast cancer cells by targeting hexokinase 2 (HK2) .Methods:Breast cancer tissues and paracancer tissues were collected and the expression levels of miR-9 and HK2 were detected. MCF-7 cells were cultured and divided into blank control group, NC group, miR-9 group, NC-siRNA group and HK2-siRNA group. The cell viability, expression levels of HK2 and cleaved caspase-3 were detected, the targeted binding of miR-9 to HK2 was verifed.Results:the expression level of miR-9 in breast cancer was lower than that in paracancer tissue (0.52±0.08 vs 1.05±0.25, t=16.685, P<0.000) , and the expression level of HK2 was higher than that in paracancer tissue (0.73±0.14 vs 0.34±0.08, t=17.587, P<0.000) , and the expression level of miR-9 was negatively correlated with HK2; the OD490 level, the expression level of HK2 and the fluorescence activity of double luciferase reporter gene containing HK2 mRNA 3 'UTR in miR-9 group were lower than those in NC group (0.58±0.09 vs 1.04±0.21, 0.51±0.08 vs 1.18±0.24, 41.11±9.28 vs 148.28±29.59, t/P=4.027/0.007, 5.297/0.002, 6.912/0.001) , and the expression level of cleaved caspase-3 was higher than that in NC group (1.08±0.26 vs 0.42±0.09, t/P=4.797/0.003) . The OD490 level and the expression level of HK in HK-siRNA group were higher than that in NC-siRNA group, and the expression level of cleaved caspase-3 was higher than that of NC-siRNA group. Conclusion:miR-9 can inhibit the proliferation of breast cancer cells, and its mechanism may be related to the targeted inhibition of HK2 expression and the increase of downstream cleaved caspase-3 expression.

Objective To summarize our experience on the food safety for residents and disaster relief workers in Li county after Wenchuan earthquake in Sichuan province.Methods A comprehensive survey was conducted to assess the current food hygiene status.According to the survey results,we had integrated the local forces and strengthened the food hygiene surveillance and quality detection focusing on the crucial procedures.Besides,effective health educations were applied to advocate the rational dietary after earthquake.Results There was no any food safety incident in the county,and the hygiene awareness of local residents has been improved.Conclusion Powerful organization,focused management and multi-collaboration are the important elements to accomplish the food safety control after earthquake.

To assess the presence of genetically modified (GM) maize and soybean in a range of commercialized feed in Shanxi province of China in 2015, improved hexadecyltrimethy ammonium bromide (CTAB) method was used to extract DNA. The screening of packed feeds was carried out by qualitative PCR. Then positive feeds were unpacked and detected by the CaMV 35S promoter, NOS terminator, zSSIIb, Lectin and CryIA (b) genes. The identified maize and soybean events were confirmed by event-specific MON810 and GTS40-3-2. Results showed that 83.3% of the feeds was tested positive for GMOs, in which positive rates of maize, soybean, pig and layer feeds were 6.67%, 100%, 93.3% and 73.3%, respectively. The results of real-time PCR were consistent with qualitative PCR. These results indicated that commercialized GM feed had a wide positive product scope in Shanxi province of China. Further studies are necessary to study effects of feeding livestock and poultry with feed containing GM ingredients on animals and their products.