BACKGROUND: Construction of lentivirus vector has the advantages of simplicity,it is regard as the most effective and successful method in transgene therapy. OBJECTIVE: To clone the metastasis suppressor gene KiSS-1 from human normal placenta tissue and construct its lentivirus vector. DESIGN,TIME AND SETTING: The open experiment was performed at the Laboratory of Fujian Normal University from September 2006 to December 2007. MATERIAL: The pNL-IRES2-EGFP vector was conservated by the Laboratory of Fujian Normal University. METHODS: Total RNA was extracted from human placenta tissue. The opening reading frame cDNA of KiSS-1 was isolated by using RT-PCR,and cloned into its lentiviral vector pNL-IRES2-EGFP to construct expression plasmid pNL-IRES2-EGFP-KiSS-1. MAIN OUTCOME MEASURES: Clone objective gene fragment of KiSS-1,restriction enzyme digestion and gene sequencing of the recombinant plasmid pNL-IRES2-EGFP-KiSS-1 were observed in the study. RESULTS: The nucleotide sequence isolated from the recombinant plasmid pNL-IRES2-EGFP-KiSS-1 was confirmed the same as expected by restriction enzyme digestion and gene sequencing. CONCLUSION: The recombinant plasmid pNL-IRES2-EGFP-KiSS-1 has been constructed successfully.

BACKGROUND: Recently, miR-378 has been shown to modulate the anti-hypoxia capacity of bone marrowmesenchymal stem cells (BMSCs) and reduce cell apoptosis under hypoxic conditions.OBJECTIVE: To investigate the benefits of miR-378-upregulated BMSC transplantation in a rat model of acutemyocardial infarction.METHODS: Primary rat BMSCs were cultured in vitro. Until passage 3, the cells were infected with the lentiviruscarrying synthetic miR-378 gene fragments. A rat model for acute myocardial infarction was constructed by ligatingthe left anterior descending artery. Thereafter, the animals were randomly assigned to three groups: control group(n=10), BMSCnull group (n=16) and BMSCmiR-378 group (n=16). In the latter two groups, 50 μL of normal salinecontaining 1×107 empty virus-transfected or miR-378-transfected BMSCs was injected into the region of myocardialinfarction, respectively. Only 50 μL of normal saline was injected in the control group. Twenty-four hours later, theapoptosis of transplanted BMSCs was evaluated with TUNEL, and expression level of vascular endothelial growthfactor and transforming growth factor-β was detected using western blot assay. Four weeks after treatment, the leftventricular function of rats was assessed by echocardiography, and then histological and molecular biology analyseswere performed.RESULTS AND CONCLUSION: At 24 hours postoperatively, there were less apoptotic BMSCs and higher expressionlevels of vascular endothelial growth factor and transforming growth factor-β in the BMSCmiR-378 group than in theBMSCnull group (n=6, P < 0.001). Four weeks later, there were more transplanted BMSCs and BMSCs-derivedcardiomyocytes in the BMSCmiR-378 group than the BMSCnull group (n=10, P < 0.001). Moreover, increased new vesseldensity (P < 0.001), decreased infarcted area (P < 0.001), preserved left ventricular ejection fraction (P < 0.05), reducedleft ventricular end-diastolic volume (P < 0.05) were found in the BMSCmiR-378 group, compared with the other two groups.The above parameters were better in the BMSCnull group than the control group (P < 0.05). Overall, the upregulation ofmiR-378 could enhance the capability of BMSCs against hypoxia, and consequently promote myocardial repair afterimplantation, providing a new strategy for cell therapy of myocardial infarction.

0.05).Conclusion Intracoronary transplantation of ABMMNCs may not improve the heart function of DCM with abnormal Q wave.

AIM:To investigate the role of JAK-STAT pathway , IL-1βand IL-6 in the PC12 cells with X-ray irradiation.METHODS:The PC12 cells were irradiated with X-ray at doses of 2, 4 and 8 Gy.After 24 h, the levels of IL-1βand IL-6 were detected by ELISA .The protein levels of p-JAK1, p-JAK2, p-STAT1, p-STAT3 and p-STAT5 were measured by Western blot .RESULTS:Compared with control group , the levels of IL-1βand IL-6 increased .The protein levels of p-JAK1, p-JAK2, p-STAT1, p-STAT3 and p-STAT5 increased with the doses of X-ray exposed.CONCLUSION:JAK-STAT signaling pathway , IL-1βand IL-6 play a role in the injury of PC12 cells with X-ray irradiation.

Objective To assess the application of ultrasonography in prenatal diagnosis of fetal choledochal cysts.Methods The clinical data of 25 fetuses,who were diagnosed as fetal choledocho cysts using ultrasonography and followed up,were retrospectively analyzed.Results Among 25 cases,24 cases were confirmed as congenital choledochal cysts after normal delivery or abortion,and 1 case of liver cyst was misdiagnosed.The 24 cases' prenatal ultrasound showed liver cystic space occupying wihich was connected to intrahepatic bile duct or gallbladder.The cysts wall was thin,and the colour Doppler flow imaging CDFI showed that blood vessels circled around the cyst.In 24 fetuses with prenatal choledochal cysts,21 were fullterm birth,3 were aborted including 1 case with other congenital abnormalities.The size of choledochal cysts was not changed in 5 cases,and was enlarged with the gestational ages in remaining 16 cases fetuses.The choledochal cysts did not affect fetal growth and development.The newborn infants with congenital choledochal cysts were treated surgically within 8 months after birth and all recovered well.Conclusion Fetal choledochal cysts have typical sonographic manifestations.Ultrasonography can be used for prenatal diagnosis of congenital choledochal cysts,which can be successfully treated by surgery early after birth.

Objective: To evaluate value of application of intravascular ultrasound imaging (IVUS) in diagnosing borderline lesions in left anterior descending artery (LAD) and left main coronary artery (LM). Methods: According to results of coronary angiography (CAG) in 60 cases with coronary borderline lesions, including 20 cases in LM, 20 cases in proximal segment of LAD and 20 cases in middle segment of LAD, the diagnostic value of IVUS in coronary borderline lesion was evaluated. Results: Compared with CAG, mean diameter stenosis rate of each coronary artery [LM: (65.3l±7.81) % vs. (75.28±8.89) %，proximal segment of LAD: (66.67±8.79) % vs. (78.89±7.88) %，middle segment of LAD: (71.55±6.83) % vs. (75.3l±7.81) %, P<0.01 all] significantly increased in IVUS. The differences of detection rate of plaque calcification and plaque rupture were no significant between CAG and IVUS（＞0.05）. Conclusion: Different degrees of underestimation of coronary artery stenosis exist in CAG, especially in proximal segment of LAD. IVUS can be an effective complement to CAG.

Objectives:To investigate the association and the prognostic significance of the expression of telomerase activity (TA) and apoptosis (APO) in tissues of primary breast cancer.Methods:TRAP-PCR and ELISA were developed to detect the expression of TA,and make quantitative and qualitative analysis respectively.TUNEL was also used to evaluate apoptosis cells in cancer lesions and count apoptosis index (AI).The expression levels of proteins of oncogene Bcl-2 and p53 were measured by means of S-P immunohistochemistry.Results:The mean TA positive expression rate was 89.8%,and its mean optic density was (A) 0.63±0.29,which were significantly higher than those of normal tissues and benign breast lesions (P<0.05).The 5 years survival rate of patients with positive TA expression was lower (60.3%) than those of negative expression (77.4%).There were no relations between TA expression and patients ages,TNM staging,lymph nodes metastasis,size,locations and pathological types of cancer.But TA was positively correlated with APO (r=0.733 1,P<0.05) and the protein expression of oncogene Bcl-2 (r=0.781 4,P<0.05) and negatively correlated with that of p53 (r=-0.625 5,P<0.05).Moreover,in patients receiving preoperative chemotherapy,the levels of TA expression were much lower than those without chemotherapy (64.7% vs 85.1%),and the former had higher AI than the latter (7.19±3.75% vs 3.23±2.04%),intraarterial chemotherapy group were superior significantly to adjuvant group (P<0.05).Multivariate Cox Regression Analysis showed that only TA and AI influenced the survival time of primary breast cancer patients,and patients with higher positive TA expression and AI>4.0% had shorter survival time than those of negative TA expression and AI<4.0% patients.Conclusions:There was a significant correlation between TA and APO and its related oncogene Bcl-2,p53.TA and APO are of great diagnostic values,therapeutic helps and prognostic significance in the formation and the development of primary breast cancer.

Hematopoietic Stem Cell Transplantation ; Histocompatibility Antigens Class I ; genetics ; Humans ; Polymorphism, Genetic ; Transplantation, Homologous

Objective To compare the difference between traditional model teaching method and the system of internal circulation system with high simulation of the teaching method of the teaching method of the arm of the arm of the intravenous infusion in children.Methods The research objects were taken as the research object,and the model of the arm of the 2013 students in Kunming college was taken as the research object,and the traditional model was the control group.The difference between the two groups were compared between the 100 groups.Results Compared with the control group,the study group's practice skill score (87.8 ± 6.9) points was higher than that in the control group (78.5 ± 7.4) (P ＜0.05);the teaching cost (2.64 ± 0.4 yuan) was higher than that in the control group (12.89 ± 0.6 yuan) (P ＜0.05);the success rate of puncture in the study group 82.8％ was higher than that in the control group (62.3％) (P ＜0.05) Conclusion The internal circulation system of high simulation of pediatric intravenous infusion arm model teaching method in basic nursing practice teaching can improve the teaching effect,the students teaching goal,and mobilize the enthusiasm of students learning.

Objective To investigate the imaging characteristics, clinical features and outcomes of penetrating ar-tery disease (PAD) cerebral infarction. Methods One hundred cases of cerebral infarction were divided into 44 cases of PAD group and 56 cases of LAA (large artery atherosclerosis ) group by brain MRI and neck CTA, or DSA neck artery ultrasound. The clinical features, imaging characteristics, outcome and progressive motor deficits(PMD) were compared between PAD cerebral infarction and large artery atherosclerosis (LAA) cerebral infarction. Results There were For-ty-four (44%) cases of PAD cerebral infarction, 56 (56%) cases of LAA cerebral infarction according to 2011 Chinese ischemic stroke subclassification (CISS). The smoking, drinking and TIA histories was significantly less in PAD group (27.27%) than in LAA group (50%) (P=0.021,0.023 and 0.025 respectively,);compared with LAA group, PMD occured in PAD group more frequently. (56.82% vs. 30.36%, P=0.008); lesions were located in the lateral ventricles in most PAD group which was significantly different from LAA group (52.27% vs. 21.43%, P=0.001). Modified Rankin Scale, (mRS) was not significantly different between two groups (1.43 ± 1.17 vs. 1.43 ± 1.45, P=0.99) at 3 months. Conclu-sions Patients with PAD cerebral infarction have PMD at early stage but have good prognosis at three months.