Objective： To clone rat cardiac myosin α he avy chain cDNA fragment encoding aa736-960 and construct its recombinant retrov irus vector(RV). Methods: The 681 bp target gene was amplified f rom heart tissue of young rats with RT-PCR, fusion gene of huIL-2/myosin was c onstructed by splicing with huIL-2 cDNA using ligation methods and its RV was constructed. RT-PCR and immunohistochemical assay were used to iden tify the expression of myosin protein in transfected cell. Results: The determination of nucleotide sequence showed that the nucleotide and ami no acid sequence of gene clone was the same as reported, its openin g reading frame was correct, the digesting result of pLNC-huIL-2-myosin was i dentical with the predicted. NIH3T3 cell was transfected with recombinant RV, and G418-resistant NIH3T3 cell was established.RT-PCR analysis indicated tha t mRNA of pLNC-huIL-2-myosin was present in cell transfected with RV. The im munohistochemical assay also showed that the myosin protein expression was highe r in the cell transfected with constructed RV. Conclusion: Rat cardiac myosin α heavy chain cDNA has been cloned and its RV has also been cons tructed and expressed in NIH3T3 successfully, it will contribute to research of prevention and treatment of heart immune injury by cardiac myosin gene transfer to induce specific tolerance.

Objective To investigate the effects of different astragalosides(AST) and hydro-soluble astragali polysaccharide(APS) of Astragalus membranaceus on inducing erythroid differentiation of human leukaemic K562 cells.Methods APS and AST were extracted by alcohol or water from Astragalus membranaceus.K562 cells were treated with APS,AST and sodium butyrate(BA) respectively.The proportion of benzidine-positive cells was examined after 1-4 days culture.MTT assay was performed for evaluating the proliferation effects of APS,AST and BA on K562 cells.Results The percentages of benzidine-positive cells induced by APS,AST and BA were 13.2%,2.9% and 17.5%,respectively.The kinetic characteristics of K562 cells induced by different levels of APS(1,2,4,8mg/ml) indicated that 4 mg/ml APS was sufficient to induce K562 cells to turn to be benzidine-positive.The results suggested that APS,other than AST,could induce the K562 cells towards erythroid differentiation.Compared with BA,the percentage of benzidine-positive cells induced by APS was lower at 24h,48h and 72h(F=237.44,P=0.00),while the total number of benzidine-positive cells was higher at 96h(F=322.25,P=0.00).The results of MTT assay for Absorbance(A) showed that APS and AST had no inhibitory effects on growth of K562 cells(P=0.28,P=0.11),while BA showed an obvious inhibitory effect on K562 cells(P=0.00).Conclusion Astragalus membranaceus has pharmacological effects to induce the K562 cells towards erythroid differentiation,and its valuable constituents are contained in hydro-soluble astragali polysaccharide(APS).The induction of differentiation is most evident with a dosage of 4 mg/ml APS.The cell growth curve reveals that APS has no inhibitory effect on K562 cells.

Objective:To clone the rat cardiac myosin ? heavy chain cDNA fragment encoding aa736 960 and construct its recombinant retrovirus vector Methods:The 681 bp target gene was amplified from heart tissue of young rats with RT PCR Fusion gene of hIL 2/myosin was constructed by splicing with preserved region of hIL 2 cDNA using ligation methods and subsequently the plasmid pLNC hIL 2 myosin was constructed NIH3T3 cells and PA317 cells were transfected with plasmid pLNC hIL 2 myosin using Lipofectamine After screening with medium containing G418, the positive clone was chosen and was detected using RT PCR, immunohistochemistry, immune electron microscope and dot blot Results:The determination of nucleotide sequence showed that the nucleotide and amino acid sequence of the gene cloned was the same as the reported sequence, and its open reading frame was correct RT PCR analysis indicated that mRNA of the fused gene was present in the positive clone Immunohistochemistry, immune electron microscope and dot blot showed that the fused gene IL 2 myosin was successfully expressed Conclusion:The fused gene of rat cardiac ? heavy chain fragment and the preserved region of human IL 2 was constructed and expressed successfully

Objective: To study shame of college student through phenomenological method. Method: 147 college students (male 53, female 94, average age 20.2) were involved. Each subject was asked to describe his/her personal shame experience, which was rate on phenomenological dimensions. Experience of Shame Scales (ESS) was used either. Result: The finding from phenomenological rating was in accord with the theory of “self orientation". Students with stronger shame proneness showed tendency to attribute negative events to him/herself. Conclusion: Shame is an acutely painful experience that involves a marked self-focus (self oriented) emotion. Shame proneness tended to have stronger sense of worthlessness and powerlessness and have much more strategies of denying and escaping in hard situations.

Objective To investigate cytochrome P450 1A2~* 1C gene polymorphism in Dai and Han nationality volunteers from Dehong autonomous prefecture in Yunnan province. Methods One hundred and seventeen Dai and 112 Han nationality volunteers from Dehong autonomous prefecture in Yunnan province were enrolled in this study. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was employed in genotyping analysis. Results There were 45 wild-type homozygotes (G/G), 63 heterozygotes (G/A) and 9 homozygotes among 117 Dai nationality volunteers, while 63 wild-type homozygotes (G/G), 44 heterozygotes (G/A) and 5 homozygots among 112 Han nationality volunteers. There was significant difference in the incidences of the genotypes between the two populations (P<0.05). The distribution of the genotypes of either population was in Hardy-Weinberg equilibrium. The frequency of the allele A in the 1ocus-2964 of CYP1 A2 was 35% (95% CI30%-40%) and 24% (95% CI 20%-30%) respectively in Dai and Han nationality volunteers. There was significant difference in the frequency between two populations (P<0.05). There was also significant difference in the frequency between Dai nationality volunteers and the populations of other regions. Conclusion CYP1A2~*1C gene polymorphism is one of factors of producing individual and racial variation in pharmacology in Dai and Han people from Dehong autonomous prefecture in Yunnan province.

Purpose:To constract and identify eukaryotic expression vector carrying human VEGF cDNA to cure human ovarian cancer with antiangiogenesis method. Methods:Reverse transcription PCR for VEGF, VEGF was inserted into eukaryotic exprssion vector pcDNA3 to construct pcDNA3-VEGF(-), in which restriction enzyme analysis was used to confirm the reverse orientation fo the VEGF cDNA from the individual transformants. The vector was transfected into human ovarian cancer cell HO-8910 and the positive clone was screened by G418,VEGF expression was confirmed by RNA dot blot. The VEGF expression of HO-8910 cells before or after transfection was detected by Western blot and imunofluorescence.The biological characteristics of HO-8910 cells before and after transfectionwas inspected. Results: VEGF gene was obtained by RT-PCR, the pcDNA3-VEGF(-) vector was obtained. VEGF expression was blocked by anisense VEGF RNA. The clone formation ability of singal cell was decreased after transfected, G 1 phase cells were increased and S Phase celIs were decreased in cell cycle. The proliferation of HO-89l0 cell was reduced. The formation rate and growth speed of xengrafted tumor slowed down. Conclusions:The successful construction of antisense VEGF eukaryotic expression vector is of significance for ovarian cancer-specific antisense gene therapy.

Objective To explore the inhibitory effect of astragalus polysaccharide (APS) on the proliferation of human erythroleukemia K562 cells and its mechanisms.Methods After K562 cells (purchased from Shanghai cell bank of chinese academy of science) were treated with different concentrations (0 mg/L,100 mg/L,200 mg/L and 400 mg/L) of APS.The influences of APS on the growth rate,doubling time and cell cycle distribution of K562 cells were observed by methyl thiazolyl tetra-zolium assay (MTF) and flow cytometry,respectively.Furthermore,the reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting assay were used to detect the expressions of Cyclin A,Cyclin B,Cyclin E and p21 gene at the mRNA and protein levels,respectively.Results MTT assay findings showed that,compared to the control group (0 mg/L APS),growth rates of K562 cells treated with 100 mg/L,200 mg/L and 400 mg/L APS decreased significantly (all P ＜ 0.01),and the doubling times lengthened significantly (all P ＜ 0.01).Flow cytometry findings revealed that,compared to the control group,the G1 phase cells in K562 cells of APS group increased significantly (P ＜0.01),while the S and G2/M phase cells decreased significantly (all P ＜ 0.01).RT-PCR and Western blotting results indicated that Cyclin B and Cyclin E expression of K562 cells at the mRNA and protein levels in the APS group were significantly lower than those of the control group(all P ＜ 0.01),whereas p21 expression was significantly enhanced at mRNA and protein levels (P ＜ 0.01),and Cyclin A expression was not significantly different at mRNA and protein levels between the 2 groups (all P ＞ 0.05).Conclusions APS could inhibit the proliferation of human erythroleukemia K562 cells.APS could inhibit the proliferation of K562 cells by down-regulating the expression of Cyclin B and Cyclin E and up-regulating the expression of p21.

Objective To retrospectively analyze the perioperative complications of posterior lumbar interbody fusion (PLIF) for recurrent lumbar disc herniation and identify potential risk factors that correlate with those complications.Methods All of 71 patients with recurrent lumbar disc herniation were treated surgically with PLIF,discharged from our department between January 2008 and December 2014.Demographic and operation data were collected and perioperative complications were recorded.We analyzed whether the clinical factors (age,gender,BMI,co-morbidity,smoking,time of recurrence,blood loss,operation segment,operation time) were in correlation with perioperative complication by univariate analysis.Then we integrated the statistically significant indicators into Logistic regression equation to determine the related risk factors for complication.Results The study group consisted of 71 cases,including 42 males and 29 females.The age was 19 to 64 years old with an average of 50.6 years old,and the average BMI was 23.6 kg/m2.26 cases had perioperative complications,while there were two or more complications in 5 patients,and no mortalities.Neurologic deterioration or neuropathic pain (10 cases,14.1％) and dural tears (6 cases,8.5％) were the most common intraoperaitive complications.The other complications included nerve root or cauda equine injury,deep or superficial wound infection,urinary tract infection,respiratory system complication,cardio-vascular complication and delirium.Univariate analysis suggested that age,gender,co-morbidity,smoking,time of recurrence,operation segment,operation time were not associated with perioperative complication.The multivariate Logistic regression analysis showed BMI and blood loss were closely related to perioperative complication.Conclusion Complications of posterior spinal fusion surgery for recurrent lumbar disc herniation are affected by many factors.The most common complications are transient neurologic deterioration or neuropathic pain and dural tears.BMI and blood loss are independent risk factors.

OBJECTIVE:To observe therapeutic efficacy and safety of S-1 capsules combined with recombinant human end-ostatin in the treatment of middle and advanced primary liver carcinoma. METHODS:Totally 94 patients with middle and advanced primary liver carcinoma in the First College of Clinical Medical Science of China Three Gorges university during Feb. 2012-Dec. 2014 were divided into combination group(48 cases)and control group(46 cases)according to random number table. Both groups were given S-1 capsules 40-60 mg orally within 30 min after breakfast and supper. Combination group additionally received Recom-binant human endostatin injection 150 mg added into 0.9%Sodium chloride injection 210 mL with portable micro pump for continu-ous pump of 120 h. A course involved 14 d treatment and 7 d interval. Short-term objective therapeutic efficacy,clinical benefit re-sponse (CBR) and ADR were evaluated after 2 courses. Disease progression time and average survival period were compared be-tween 2 groups. RESULTS:Objective response rate,disease control rate,disease progression time and average survival period of combination group were 14.6%,66.7%,(5.5 ± 1.3) months,(10.7 ± 3.8) months;those of control group were 8.7%,45.6%, (4.8±1.2)months,(8.9±3.3)months,with statistical significance between 2 groups(P<0.05). CBR rate of combination group (79.2%)was significantly higher than control group(52.2%),with statistical significance(P<0.05). There was no statistical sig-nificance in the incidence of ADR between 2 groups (P>0.05). CONCLUSIONS:S-1 combined with recombinant human end-ostatin show good therapeutic efficacy and tolerance for patients with middle and advanced primary liver carcinoma,and do not in-crease the incidence of ADR.

Objective To evaluate the efifcacy and safety of Xiyanping in the treatment of hand-foot and mouth disease. Methods Based on the principles and methods of Cochrane systematic reviews, Cochrane Library, PubMed, Medline, Chinese Bio-medicine Database (CBM), China Journal Full-text Database (CNKI), VIP database and Wanfang database were searched. All of the randomized controlled trials (RCT) of Xiyanping versus ribavirin were included. The data were extracted and evalu-ated by two reviewers independently. Risk assessment tool was used to assess the risk of bias and software Revman5.0 was used for meta-analysis. Results Twenty-four RCT and 3314 patients were included. Comparing to ribavirin, Xiyanping showed better therapeutic outcomes regarding to total effective rate, durations of fever and rash elimination (RR=1.17, 95%CI:1.12~1.23;MD=-1.56, 95%CI:-2.10~-1.02;MD=-1.41, 95%CI:-1.90~-0.93). Side effects were rare in both groups and could be recovered after drug withdrawal. Conclusions The current evidence suggests that Xiyanping is superior to ribavirin in the treatment of hand-foot and mouth disease.