Objective To construct the eukaryotic expression vector of human telomerase RNA component ( hTR) and study its biological function tentatively .Methods hTR Gene was obtained by PCR from cDNA template , which was reverse transcribed from 293T mRNA and cloned into pCDNA3.0 vector.The recombinant plasmid and empty vector were trans-fected into 293T cells, and hTR expression was identified by qRT-PCR.HepG2 cells that stably transfected with pCDNA3.0-hTR were constructed and identified by qRT-PCR.These cells were used to assess the interaction of hTR with human telomerase revese transcriptase ( hTERT ) and dyskerin .Telomerase activity was also detected in HepG 2 cells transfected with pCDNA3.0-hTR.Results pCDNA3.0-hTR eukaryotic expression vector was successfully constructed by double digestion identification .The inserted fragment was confirmed by sequencing .The expression of hTR in human 293T cells and HepG2 pCDNA3.0-hTR stable cell line was identified.In addition, qRT-PCR and Western blotting results showed that hTR could interact with hTERT and dyskerin , while hTR overexpression could not regulate the telomerase activity in HepG2 cells.Conclusion The eukaryotic expression vector of pCDNA 3.0-hTR is successfully constructed and expressed.This study will contribute to the further study of cancer therapy targeting hTR .

Objective To summarize the preservation measures of the donor's heart and lung, and the postoperative immunotherapy, as well as the clinical experience of discrimination and management for graft rejection.Methods The clinical data of 2 cases of heart-lung transplantation in our department were retrospectively analyzed. Two different protective liquids were used for donor's lung lavage of 2 cases: Perfadx solution (1000 mL containing tris 0.3 mL and ilomedin 25 μg); Euro Collins solution (1000 mL containing tris 0.3 mL and PGE1 100 μg). UW solution was used for donor's heart lavage. Surgical procedure for heart-lung transplantation was classic technique in situ. The schedule of immunosuppression was induced by Basiliximab, and combined with cyclosporine+ mycophemolate mofeil+corcal hommone after operation. recipient's blood count, organ's functions, the sizes of every cavity of heart, IVSPW and LVPW were observed during early post-operation. The recipients were subjected to chest CT scan, fiberoptic bronchoscope and tissue pathological study when necessary to find the signs of rejection promptly. When the rejection occurred in the recipient, cortical hormone's impulse therapy was given and the dose of immunosuppression was adjusted in time.Results Two patients discharged in 80 days and 141 days after operation. The patients were followed up for 54 months and 50 months respectively, and their life qualities were very well. Acute rejections occurred on the 10th and 26th day in one case, and in another case, acute rejections occurred on the 29th and 87th day after operation. All were conversed by cortical hormone's impulse therapy and adjusting the dose of immunosuppressants. When acute rejection occurred, the blood count had significant change, and IVSPW and LVPW were increases. They were returned the normal range after corresponding therapy.Conclusion Perfidx solution and Euro-Collin solution may play good protective roles for donor's lungs. UW solution may play good a protective role for donor's heart. To discriminate the clinical graft rejection and infection in time and administrate correct management will have large benefits for the patients' rehabilitation.

Objective To explore the advantages and disadvantages of ultrasound guided puncture microwave ablation and traditional surgical treatment of nodular goiter.Methods From January 2014 to June 2015,102 cases of nodular goiter who were treated in the Traditional Chinese Medicine Hospital of Gaozhou Affiliated to Guangzhou University of Traditional Chinese Medicine were randomly divided into two groups,51 cases in each group.Respectively by ultrasound guided puncture nodules of thyroid swollen microwave ablation (microwave ablation group) and traditional operation type removed most of the thyroid nodules (traditional surgery group) in the treatment of containing.The amount of bleeding,operation time,hospital stay,medical cost,clinical efficacy,postoperative complications and the degree of pain,and thyroid function were compared between the two groups.Results The amount of bleeding,operation time and length of hospital stay in the microwave ablation group were better than those in the traditional operation group((2.02±0.79) ml vs.(28.24±18.49) ml,(34.20 ±9.60) min vs.(75.59±25.41) min,(3.55± 1.05) d vs.(10.27±2.68) d),the medical cost of the microwave ablation group was higher than that of the traditional operation group((14 341±593) yuan vs.(5 812±970) yuan),the differences were statistically significant (t =-10.12,-10.88,-16.68,53.54;P ＜0.01).There were significant difference in terms of the postoperative pain and complications between the groups (x2=50.86,13.22;P＜0.01).The total absorption rate was 100％ in 18 months after the ablation of the nodule.There was no significant difference about postoperative residual small nodules between the two groups(P ＞0.05).Conclusion Compared with the traditional treatment,the microwave ablation treatment of nodular goiter with less complications,is a new technique of minimally invasive,beauty,safe and feasible,the exact clinical effect,and worthy of popularization and application.

Objective To explore the clinical feature of severe enterovirus 71 (EV71) associated hand foot and mouth disease (HFMD),and genotype of EV71.Methods Fluorescent quantitation PCR was done for detecting EV71.RT-PCR was performed to amplify VP1 for sequencing and identifying genotype.A retrospective analysis was performed based on the clinical data of 15 cases with severe EV71 infection.Results EV71 nucleotide was positive in all 15 cases.The genotype of EV71 was C4.All cases had abnormal temperature and followed with nervous symptoms in the early stage.Average time was 1.26 days from onset to severe complications appearance.Eleven cases progressed to neurogenic pulmonary edema.Four cases accepted nasal continuous positive airway pressure.Eleven cases accepted oral trachea cannula mechanical ventilation.Except for 3 cases died,one case abandoned,others 11 cases were cured.Conclusion The isolated strains of EV71 in this study are all C4 genotype.All cases with severe EV71 infection were followed with nervous symptoms in the early stage,most of whom would progress to neurogenic pulmonary edema.The mortality would be cut down by using mechanical ventilation in early stage.

Objective To investigate 13 high-risk types of HPV (HR HPV) infection rates in women with different grades of cervical lesions.Methods A total of 350 women, who were hospitalized in the department of gynecology in Bao′an Maternity & Child health hospital, were enrolled for the study.TCT technology was used to evaluate the cervical epithelium.The group were divided according to the cytology results.Multiplex real time PCR (mRT PCR) was used to detect the viral loads.HR HPV infection rate of different groups were analyzed using χ2 test or Fisher exact test.HR HPV viral loads of patients in different grades of cervical lesion groups were compared using Kruskal-Wallis or Wilcoxon test, and the age distribution of HR HPV positive group and negative group was analyzed by using Wilcoxon test.Results The HR HPV infection rates of NILM, ASCUS, LSIL, HSIL were 3.4% (10/295), 20.0% (7/35), 78.6% (11/14) and 100.0% (6/6), respectively.HR HPV positivity in NILM was lower than ASCUS (χ2=14.43,P<0.01) and LSIL (χ2=107.69,P<0.01), HR HPV positivity in ASCUS was lower than LSIL (χ2=14.76,P<0.01). The median of HR HPV viral loads in NILM, ASCUS, LSIL and HSIL were 4.10 (3.38-6.27), 5.33 (3.63-6.66), 5.77 (4.01-7.01) and 5.58 (4.19-5.85) respectively (copies/ml,lg).Combined ASCUS, LSIL and HSIL groups into cervical lesion group, HR HPV viral load of which was higher than that of NILM (U=43.0, P<0.05).The median Ages of HR HPV positive group and negative group were 36 and 33, respectively.No statistical significance was found between them (U=4 544, P>0.05).Conclusions The present study revealed that HR HPV infection was related to cervical lesion, but there was no correlation between viral load and cervical lesion grade. In additional, no difference in age distribution was found between HR HPV positive group and negative group.

Objective To investigate the features of lipid ratios in patients with newly diagnosed T2DM, and the effects of intensive insulin treatment on them. Methods 90 patients with newly diagnosed T2DM and 58 matched people with normal glucose were enrolled to assess height,weight,waist circumference,blood glucose and lipid profiles. BMI,TC/HDL-C,TG/HDL-C,log(TG/HDL-C),LDL-C/HDL-C,HOMA-B and HOMA-IR were calculated respectively. All the patients received the continuous subcutaneous insulin infusion with insulin pump. The treatment continued for more 10~14 days after blood glucose reached the standard. All the above indi-cators were reexamined after treatment. Results Dyslipidemia in patients with newly diagnosed T2DM mainly showed as hypertriglyceridemia and decreased HDL-C compared to the control group(P<0.05). TC/HDL-C,TG/HDL-C,log(TG/HDL-C)and LDL-C/HDL-C significantly increased in these patients(P<0.01). After short-term intensive insulin therapy,all lipid ratios were significantly decreased and the changes of lipid ratios were positively correlated with the change of HOMA-IR(P<0.05). Conclusion Short-term intensive insulin therapy for patients with newly diagnosed type 2 diabetes can significantly lower the lipid ratios related to HDL-C. The effects may be closely related to improvement of insulin resistance.

Objective To analyze the fetus chromosome abnormalities in women with advanced maternal age under the universal two-child policy. Methods A total of 10607 women underwent prenatal diagnosis proce-dures from the year of 2015 to 2016,among which 3569 cases were with advanced maternal age. Cytogenetic karyotyping was conducted with chromosomal microarray analysis(CMA)in 854 cases. The ration of chromosome abnormalities in the two population groups(aged from 35~39 and over 40)were counted. The type of abnormali-ties were also analyzed. Results The ration of women with advanced maternal age in 10607 cases underwent pre-natal diagnosis between 2015 and 2016 was 33.6%. The respective rations were 27.2%and 37.9%. The detection rates of chromosome abnormalities were 7.9% and 10.8%,with significantly difference. The significant differences were also found in the detection rate of chromosome aneuploidies ,but no differences in other chromosome abnor- malities. CMA was benefit to detect the micro chromosome abnormalities. Conclusions In 2016,the number of prenatal diagnosis procedures increased under the two-child policy. The detection rate of chromosome abnormalities also significantly increased. The incidence of chromosome abnormalities was higher with the growing age of preg-nant women. Genetic counseling must be presented and prenatal diagnosis should be promoted in women with ad-vanced age.

Objective To investigate the specimen source and gene phenotype of ESBLs in ESBLs-producing Klebsiella pneumonia of people′s hospital of Sanya city,so as to provide basis for clinical use of drugs and nosocomial infection.Methods Klebsiella pneumoniae was isolated from specimens during January 2013 to December 2014,bacteria identification and susceptibility tests were detected by Phoenix-100 system biochemical,supplementary susceptibility test was confined by K-B method according to 2014 CLSI standards.WHONET 5.6was used in the statistical analysis of all data.Results Totally 213 strains Klebsiella pneumoniae were isolated.The detection rates were 78.4％ of the respiratory secretions,8.92％ and 5.2％ respectively of the secretion and the midstream urine.The strains had a certain resistance to commonly used antimicrobial.The highest resistance rate was 98.1％ to cefotaxime,and the lowest resistance rate was 2.86％ to imipenem.There were 195 in 213 ESBLs producing Klebsiella pneumoniae strain were detect one or more drug resistance gene.The detecting rates of 6 p-lactamase gene of CMY,CTX,TEM,SHV,DHA1 and KPC were 6.10％,76.53％,59.62％,76.06％,12.21％ and 2.82％.Conclusion Klebsiella pneumoniae is mainly isolated from respiratory secretions in the hospital,has a certain resistance to commonly used antimicrobial.We should learn more about the distribution of resistance genes of ESBLs strains,improve the efficiency of the treatment of the infection and to control nosocomial infection and the incidence of multi-drug resistance.

Objective To compare real time PCR with PCR-reverse dot blot hybridization (PCR-RDB)for detecting human pap-illomavirus (HPV)infection in women.Methods A total of 109 genital specimens from women were collected in the study.All specimens were tested HPV by using real time PCR and PCR-RDB,discrepant samples were tested again by PCR-xMAP.Results The concordant rate was 83.5%(91/109)between real time PCR and PCR-RDB (kappa=0.671),the other 18 discrepant samples were retested by PCR-xMAP,7 of those were identical with real time PCR and 11 with PCR-RDB.No differences of PCR-RDB pos-itive rates were found between the high and low viral load groups (χ2 =1.476,P =0.224).Conclusion It demonstrated moderate consistency between real time PCR and PCR-RDB.The HPV positive rates of PCR-RDB were stable when the viral loads were 103-108 .

Objective To investigate the female human papillomavirus(HPV)infection situation in Baoan district,the HPV pos-itive rates in different age groups and the subtypes distribution.Methods PCR followed with reverse dot blot was performed to ex-amine 23 kinds of HPV genotypes in 2 627 female patients in our hospital from the January 2011 to December 2012.Results In 2 627 samples,the positive rate of HPV was 23.94% (629 cases),in which the infection rate of single low-risk type was 15.1%(95 cases),the main HPV genotype was HPV43 (7.79%);the infection rate of high-risk type was 55.17% (347 cases),the 3 most prevalent HPV genotypes were HPV52 (12.56%),HPV16 (9.86%)and HPV58 (7.79%).The multiple HPV infection ac-counted for 29.73% (187 cases).The HPV infection rates in different age groups were 50.0% in age 15-20 years,24.7% in age 21-30 years,20.8% in age 31-40 years,25.8% in age 41 -50 years,42.1% in age >50 years respectively,the differences had statistical significance.Conclusion The HPV infection rate is 23.94% in Bao′an district.The most prevalent HPV genotypes are HPV 52,16,58,43.Women in age 15-20 years old have a higher infection rate.