Background and purpose:Apollon gene is highly expressed in leukemia and other tumors. The study aimed to discuss whether RNAi technology can reverse multidrug resistance of chronic myeloid leukemia cell line K562 through constructing a eukaryotic vector of short hairpin RNA (shRNA) targeting at Apollon gene. Methods:The eukaryotic vector pGPHI-GFP-Neo-Apollon with shRNA targeting at Apollon gene was constructed and then transfected into K562 cells by LipofectamineTM2000, and G418 pressure selection. Reverse transcription-polymerase chain reaction (RT-PCR) and immunolfuorescence were used to detect the expression of Apollon mRNA and protein after Apollon was transfected stably in K562 cells. The changes of sensitivity of K562 cells to leurocristine (VCR) and etoposide (VP16) after transfection with shRNA-Apollon were detected by MTT method, and the apoptosis rate was detected by flow cytometry. Results: pGPHI-GFP-Neo-Apollon carrier was constructed successfully and expressed stably in K562 cells, and after G418 screening, it silenced Apollon mRNA and protein expression effectively. According to the result of MTT, the sensitivity of K562 cells to VCR and VP16 increased significantly in the group of gene interference, with half of its inhibition concentration (half-inhibitory, IC50) value signiifcantly lower than the control group (P<0.05);Flow cytometry showed that the cell apoptosis rate was increased signiifcantly (P<0.05), but there was no statistically signiifcant difference in the apoptosis rate between shRNA negative control group and normal control group (P>0.05). Conclusion:pGPHI-GFP-Neo-Apollon carrier can enhance the abilities of VCR and VP16 to induce the apoptosis of K562 cells, namely an increase of sensitivity to these chemotherapeutics in K562 cells, it is hinted that RNA interference targeting Apollon gene may reverse the multidrug resistance of leukemia cells in some degree.

OBJECTIVE To offer scientific basis for reasonable usage and management of antibiotics through surveying the situation and problems of antibiotics usage in surgery during perioperative period.METHODS The antibiotics usage status among 462 cases of patients during perioperative period was retrospectively analyzed.RESULTS Antibiotics were used in 460 patients.The rate of prophylactic usage was 83.26%,the rate of therapeutic one was 16.74%;the duration of postoperative prophylaxis was:80.68% of the patients were treated with antibiotics for 4 days or more,the longest one was 28 days;43.26% of the patients received single antibiotic treatment,and 44.13%,11.96%,or 0.65% of patients received 2,3 or 4 kinds of antibiotics combined therapy respectively.CONCLUSIONS Improper application of antibiotics during perioperative period needs higher level administration.

OBJECTIVE To investigate the risk factors of professional exposure among medical staff and explore the preventive strategies.METHODS The influence of factors including profession,exposure process,exposed materials,treatment process,and follow-up among 110 medical staff with professional exposure was analyzed.RESULTS The general incidence of professional exposure was 52.73%,35.45% and 6.36% for clinical doctors,nures,and workers,respectively.Among all exposure cases,we found the incidence was 62.73% for pricking wound,20% for various liquid contamination,4.55% for glass injury,2.73% for scalpel cutting,and 4.55% for other instrument related damage.The reasons leading to exposure were treatment and nursing care,surgery assistance,and instrument and materials manipulation,which accounted for 40%,31.82%,and 18.18% of total incidence,respectively.All of the exposure cases had been properly treated.None of the investigated cases had been found illed with blood derived transmitted disease caused by professional exposure.CONCLUSIONS It is critical to intensify the training of medical staff for the knowledge of professional exposure and to improve their personal protective awareness,so that the incidence of professional exposure could be reduced.

OBJECTIVE To investigate the antibiotic use during perioperative period in surgery after administration according to Antibiotic Drug Use Guideline.METHODS The antibiotics usage status among 462 outpatient cases in Mar 2005 and 483 cases in Mar 2006 during perioperative period in surgery were surveyed.RESULTS The preventive use rate of antibiotic medication in surgery in 2005 and 2006 were respectively 99.48% and 88.12%(P

Objective To screen siRNAs that can effectively inhibit Apollon gene expression and determine the cellular functions of those siRNAs.Methods A chemical synthesis method was used to synthesize 3 siRNA sequences against different sites of Apollon.They were transfected into the human breast cancer MCF-7 cells by using Lipofectamine 2000.mRNA level of Apollon was determined by reverse transcription-polymerase chain reaction (RT-PCR).Cellular immunity fluorescence quantitative analysis combined with confocal laser technology was used to determine the protein level of Apollon.Methyl thiazolyl tetrazolium bromide (MTT) assay and flow cytometry were used to determine the effects of siRNA targeting Apollon on proliferation and apoptosis of MCF-7 cells,respectively.Results Three pairs of siRNA could significantly inhibit Apollon mRNA expression,at the inhibition rates of (36.201±11.629) ％,(67.308±7.686) ％and (47.123±12.000) ％,respectively (P ＜ 0.05).After tranfection by siRNA2,Apollon protein fluorescence intensity was (14.97±2.08) ％ compared with control cells.The cell proliferation MCF-7 was inhibited by (73.361±2.118) ％and apoptosis was increased by (28.793±0.743) ％.Conclusions Screened siRNA2 effectively silences Apollon gene expression,effectively inhibits the proliferation and increases the apoptosis of MCF-7 cells.This provids the foundation for its clinical application in cancer therapy.

Objective To investigate the evaluation of cardiopulmonary exercise test (CPET) in the patients with chronic obstructive pulmonary disease (COPD), and the relationship with the pulmonary function test (PFT). Methods 26 COPD patients and 30 normal controls were examined with CPET and PFT. Results The parameters of CPET, including maximum oxygen uptake (VO2max), anaerobic threshold (AT), maximum minute ventilation (VEmax) and breathing reserve (BR) were lower in COPD group than those in control group (P<0.05). There were no significant differences in Wmax, oxygen pulse (O2pulse) between two groups (P>0.05). VO2max, AT, VEmax and BR were positively correlated with forced expiratory volume in one second (FEV1) and peak expiratory flow (PEF) in COPD group （P<0.05）, but were not correlated with forced vital capacity (FVC) and FEV1/FVC (P>0.05).Conclusion The pulmonary function of COPD patients can be evaluated with CPET, especially VO2max, AT, VEmax and BR.

Ninety- five cases(49 male .46 female)with first heamorrhagic cerebral apoplexy. fromOct.1 . 1992 to Dec.31 .1993. were analyzed.The Average age was58.9. Patients were divided intotwo groups. the self care and the non- self care. Ninteen variables which were supposed likely to be relatedto functional outcome were chosed from the admitted record of each patient and analyzed with mutltiple lo-gistic regression. Results showed that the major predictors of poor outcome are coma. incontinent and hemi-plegia at onset and abnormal blood urea nitrogen. pulmonary infection and bleeding amount more than 3 0ml at admitted.Cases older than 70 years.high blood presure longer than 20 years have a tendency of moredependence in functional abilities.Factors that do not predict outcome after heamorrhagic stroke are sex.days in the hospital,cormorbidities. as well as level of blood surgar and cholestral.EKG changes at admitted time.

Objective To explored high-risk HPV genotyping PCR testing whether as a feasible means for the early screening of cervical cancer and precancerous lesions. Methods From January 2013 to June 2014, 15 192 outpatients in China-Japan Friendship Hospital voluntary were tested by high-risk type HPV genotyping PCR. The average age of them were (33±8) years old. High-risk HPV types genotyping PCR tested by fluorescence PCR technology,in which 13 kinds of high-risk HPV subtypes were detected,
including HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68. A total of 4 315 cases of them were tested by the liquid-based cytology (LCT), among them with positive of high-risk HPV genotyping tested by PCR (n=2 366) were biopsy under colposcope (648 cases) in those LCT results were positive or LCT negative but HPV16 positive or LCT negative but had the clear clinical symptoms or and non-HPV16 positive but with clear clinical symptoms. (1) Analysis high-risk HPV infection status of 15 192 women.(2)As the pathological diagnosis was the gold standard in the diagnosis of cervical lesions, analysis of the relationship among high-risk HPV infection,virus loads and cervical lesions. (3) To evaluated the value of high-risk HPV genotyping PCR tested method in screening of cervical cancer and precancerous lesions. Results ⑴ Of 15 192 cases tested by high-risk HPV genotyping PCR, 2 366 cases were HPV positive (HPV infection), the overall infection rate was 15.57%(2 366/15 192), in which a single subtype of HPV infection in 1 767 cases, infection rate was 11.63%(1 767/15 192), and multiple subtypes of HPV infection (two and more subtypes HPV infection) in 599 cases, infection rate was 3.94%(599/15 192). The HPV16, 52 and 58 infections were the most common HPV subtypes in 13 subtypes, the infection rate was 3.95% (600/15 192), 2.86%(435/15 192) and 2.67% (406/15 192), respectively. (2) The most relevant subtypes with cervical intraepithelial neoplasia (CIN)Ⅱand even higher lesion were HPV16, 52 and 58, accounted for 57.7%(154/267) of all above CINⅡlesions. The most relevant subtype with the cervical glandular intraepithelial neoplasia (CGIN) Ⅱ or above lesions was HPV18, 3 cases with CGIN Ⅱ or above lesions were all single HPV18 infection. The pathologic examination positive percentage of patients which HPV virus loads≤103 copys/104 cells was 18.2%(25/137), while the pathologic examination positive proportion was 33.3%(247/742) which HPV virus loads≥104 copys/104 cells , there was statistically significant difference between them (χ2=27.06, P=0.000).(3)Sensitivity, specificity, positive predictive value and negative predictive value for detection of CINⅡ or above using HPV genotyping PCR were 96.11%, 85.76%, 30.94% and 99.70%, respectively. Conclusions There were a guiding significance for high-risk HPV genotyping PCR tested in screening of cervical cancer and precancerous lesion. HPV16, 52 and 58 were related to the severe cervical squamous epithelial lesions, while HPV18 was related to cervical severe glandular cell pathological changes. HPV genotyping is feasible and economical as the first choice of opportunistic screening in tertiary hospitals.

OBJECTIVE:To compare the efficacy and safety of methotrexate with different methods of administrations com-bined with hysteroscopy in the treatment of scar pregnancy(CSP). METHODS:Data of 56 CSP patients were retrospectively ana-lyzed and divided into group A(26 cases)and group B(30 cases)by different methods of administrations. The fluid in group A was injected methotrexate 50 mg/m2 into the capsule,once a week. After 1 week,if the β-HCG was decreased less than or equal to 50%,the patient was additionally injected methotrexate;and if the β-HCG was decreased more than 50%,no more methotrexate was given,the lesion electrosurgical excision was conducted under hysteroscopy when β-HCG<1 000 mU/ml. Group B was inject-ed methotrexate 50 mg/m2 into muscle,once a week. After 1 week,if β-HCG was decreased less than or equal to 50%,the patient was additionally injected methotrexate;and if the β-HCG was decreased more than 50%,no more methotrexate was given,the le-sion electrosurgical excision was conducted under hysteroscopy when β-HCG<1 000 mU/ml. The clinic data was observed,includ-ing success rate,β-HCG before and 4,7,10 and 14 d after treatment and the mass diameter of CSP before and 7 and 14 d after treatment in 2 groups. The incidence of adverse reactions was recorded. RESULTS:There were no significant differences in the suc-cess rate,mass diameter of CSP before and after treatment between 2 groups (P>0.05). After treatment,the β-HCG in 2 groups was significantly lower than before,14 d<10 d<7 d<4 d,and group A was lower than group B,the differences were statistically significant(P<0.05). The incidence of adverse reactions in group A was significantly lower than group B,the difference was statis-tically significant(P<0.05). CONCLUSIONS:Methotrexate with different methods of administrations combined with hysteroscopy has good efficacy in the treatment of CSP,however,the safety of intracapsular injection is better than intramuscular injection.