Objective To construct and identify the recombinant retroviral vectors expressing siRNA for MDR1 gene knock-down,sequentially to detect the reversing effect of RNAi on the multidrug resistant phenotype in MCF-7/ADR cells. Methods The target sequences of siRNAs for MDR1 knock-down were designed by IDTdna online. The DNA sequences containing a palindrome structure were synthesized and annealed to form double-stands which were inserted into linear pSUPER vectors later. The recombinant vectors were identified by agarose gel electrophoresis after cleaving of double enzymes and by sequencing,then were transfected into 293T packaging cells which produced retroviral particles in supernatant,the retrovirus soup was used to infect MCF-7/ADR cells. The expression level of MDR1 was detected from both mRNA level and protein level by RT-PCR and Western blotting,respectively. Results The recombinant pSUPER vectors expressing siRNA were confirmed by agarose gel electrophoresis after cleaving of double enzymes and by sequence analysis. The inserted oligonucleatide sequences were identical to the original sequences and in corresponding position. Compared with control group,the inhibitory effects of pSUPER.retro-MDR1 and pSUPER.retro-MDR2 were significant. Conclusion pSUPER recombinant retroviral vectors expressing siRNA for MDR1 knock-down is construted successfully,and it has inhibitory effect on the expression of MDR1 gene in MCF-7/ADR cells.

Desmoplastic small round cell tumors (DSRCTs) are a rare malignancy found in male adolescents that initially occur mostly in the abdominal cavity. Diagnosis is based on the histologic analysis of biopsies, which typically show small round blue cells in nests separated by abundant desmoplastic stroma. DSRCTs are associated with a unique chromosomal translocation t (11:22) (p 13;q 12) that involves the Ewing's sarcoma (EWS) gene and the Wilms' tumor (WT1) gene. Reverse transcriptase-polymerase chain reaction can be used to detect the fusion gene in fresh or paraffin-embedded tissues, which confirms the diagnosis. The prognosis is particularly poor. The median survival ranges from 17 to 25 months. Management of DSRCT remains challenging despite the use of aggressive ther-apies such as polychemotherapy, debulking surgery, and whole abdominal radiation. Several methods for improving patient survival are being evaluated, such as the addition of chemotherapy and targeted therapies to normal neoadjuvant protocols, complete surgical resec-tion with hyperthermic intraperitoneal chemotherapy, postoperative intensity-modulated radiation therapy, and yttrium-90 microsphere liver embolization for treating hepatic metastases.

Objective To understand the effects of stove-improvement measures in coal-burning-induced fluoride poisoning area in Luoyang.Methods Dynamic monitoring on the fluoride concentration in indoor air,grain(corn and wheat),vegetable(chili,radish and Chinese cabbage),and the urine fluoride concentration and prevalence of dental fluorosis among8-12year old children was conducted after the performance of stove-improvement measures in coal-burning-induced fluoride poisoning area,Jiangzhuang village,Xinan county,Luoyang,during1991-2000,while the obtained data were compared with those in1990.Re sults After the performance of stove-improvement measures,all of the fluoride contents in indoor air,grain and veg-etable,the urine fluoride concentration and the prevalence of dental fluorosis of8-12aged children revealed a trend of decrease on the whole,and accorded with the corresponding standards for non-fluoride poinsoning area,despite of the fluctuating data in different years.Conclu sion The stove-improvement measures could effectively prevent the indoor air,grain and vegetable from fluoride pollution,and control the prevalence of fluoride poisoning in coal-burning-induced fluoride poisoning area.

OBJECTIVE:To assess the effectiveness of 1,6—diphosphate fructose(FDP)in the treatment of heart failure(HF).METHODS:Biomedical databases,including Medline,Embase,Cochrane Controlled Trials Register,CBM and CNKI were searched.Clinical trials were screened according to the inclusion criteria formulated beforehand,quality assessments and meta anylysis of included studies were carried out.RESULTS:14 randomized controlled trial were included,and all of them were carried out in China.Jadad score of the studies were all 1.None of the 14 trials carried out by follow-up domization for long-term effect evulation Meta-analysis of 14 trials showed that FDP can increase effective rate in the treatment of HF,and OR(95%CI)was 3.79(2.58,5.57).5 studies showed that FDP can increase cardic index,and WMD(95%CI)was 0.40(0.34,0.46).6 studies showed that FDP can increase ejection fraction(EF)with WMD(95%CI)8.31(5.51,11.11).4 studies showed that FDP can decrease left ventricular end systolic and diastolic diameter,and WMD was—3.25(—4.74,—1.76)and—3.75(—6.94,—0.56).No long-term effects were reported by the included studies.CONCLUSIONS:According to present evidence,FDP can increase the treatment effective rate when added to the basic treatment of HF.FDP can improve heart function and lefe ventricular construction showed by the indicators.However,the poor quality of the studies decreased the persuasion of the results.Large-scale and well-designed RCTs with enough follow-ups should be carried out to provide further evidence for the use of FDP.

Objective The relationship among angiotensin converting enzyme(ACE) gene polymorphism,angiotensin converting enzyme activity and coronary heart disease(CHD) was studied.Method Teh polymerase chain reaction was used to detect an insertion/deletion(I/D) of ACE gene polymorphism in the sixteen intron,then genotype and allele freqencies were counted .The method of enzyme coupling ratio was used to detect ACE activity .Results In 51 case with CHD ,the DD genotype and D allele freqencies of ACE gene were 35% and 61% respectively ,ACE average activity was (350 3?91 1)su/L.In 83 normal control subject,they were 16% and 45% and (286 7?79 6)su/L respectively.The freqencies of DD genotype and D allele and ACE average activity had significant difference.Conclusion The deletion polymorphism of ACE gene might be an latent risk factor.The higher ACE activity might play an important role in the development of CHD.

Objective To study the ratios between several pairs of endogenous androgens in morning urine samples from male and female athletes, so as to define their reference ranges.Also to study the effects of large loading and strengthening exercises on these ratios,which may give a halp to eliminating the misjudgement caused by stimulant misuses.Methods The steroid hormones were detected by GC-MS method through the American made HP5890 and HP5971 GC-MS,with methyl testosterone as an internal standard.The hormone concentrations were calculated by means of integral analysis to each specific ionic peak.Results The ratios An-Etio,5?/5? and T/ET in male athletes were all significantly higher than those in females(P

Objective To investigate the effects of repeated computed tomography (CT) scan on hematological and immunological index in mice,and to explore the related mechanism.Methods A total of 60 BALB/c mice were randomly divided into six groups as control,once,twice,4,6 and 8 times of CT scans,and administered with whole-body scan by 64-slice spiral CT with the pelvis scan mode.Thermoluminescence dosimeter was used to detect the absorbed doses of CT scan,while DNA damage (γ-H2AX),apoptosis,autophagy,cell cycle and percentages of CD4/CD8 in peripheral lymphocytes and bone marrow cells were analyzed by flow cytometry.γ-H2AX was detected 1 h after the last CT scan,and the other indexes were detected 24 h after the last CT scan.Results γ-H2AX increased after more than twice CT scans in peripheral lymphocytes and after than 4 times of CT scan in bone marrow cells (P ＜ 0.05).Apoptosis increased in peripheral blood lymphocytes and decreased in bone marrow cells after more than 4 times of CT scan (P ＜ 0.05),suggesting that lymphocytes were more sensitive while different cell subpopulation in bone marrow showed different sensitivity and response phase.No obvious changes of autophagy level were observed in peripheral blood lymphocytes,while autophagy increased in bone marrow cells after single CT scan (F =81.10,P ＜ 0.05).8 times of CT scan significantly increased the percentages of cells at G0/G1 phase and G2/M phase(x2 =11.29,28.57,P ＜0.05).The percentage of CD4 + T,CD8 + T cells decreased after CT scan,but no effects on the ratio of CD4 +/CD8 +.Conclusions Repeated CT scans could induce injury effects on hematopoietic system and immune system,and then the apoptosis.Apoptosis,autophagy and cell cycle arrest might be initiated to maintain genome stability but there is no influence on immune balance.

ObjectiveTo investigate the significance of hearing screening combined with gene screening for neonates with high-risk of hearing impairment.MethodsNeonates admitted to the Neonatal Department of Guangdong Women and Children Hospital between July 2013 and June 2014 were enrolled in this study. They were divided into high-risk group (with high-risk for hearing impairment) (n=3 129), and control group (n=5 106). Neonate hearing screening was carried out using otoacoustic emission and automated auditory brainstem response. Blood samples were collected using a standard protocol for detecting the mutations of four common deafness genes, includingGJB2,GJB3,SLC26A4 and mitochondrial 12s rRNA.Chi-square test was used to compare the differences of the pass rate of hearing screening and positive rate of gene mutations between the two groups.ResultsThe rates of failure on otoacoustic emission, automated auditory brainstem response or both in the high-risk group were 11.92% (373/3 129), 10.32% (323/3 129) and 4.83% (151/3 129), respectively, higher than those in the control group [5.03%(257/5 106), 6.56%(335/5 106) and 2.02% (103/5 106)] (χ2=130.265, 37.354 and 51.196, allP=0.000). In the high-risk group, the overall positive rate of gene mutations was 5.63% (176/3 129), and theGJB2 andSLC26A4 gene mutation rates were 3.04% (95/3 129) and 2.40% (75/3 129)], all higher than the control group [3.15% (161/5 106), 2.04% (104/5 106) and 1.06% (54/5106)] (χ2=30.301, 8.216 and 22.517, allP<0.01). But the mitochondria 12S rRNA gene andGJB3 gene mutation rates were the same in high-risk group and control group [0.19% (6/3 129) vs 0.06% (3/5 106); 0.03% (1/3 129) vs 0.00%(0/5 106), bothP>0.05]. The rates of failure on otoacoustic emission and automated auditory brainstem response of the neonates with deafness gene mutations were 9.50% (32/337) and 10.39% (35/337), respectively, higher than the neonates without [1.14% (90/7 898) and 1.29% (102/7 898)] (χ2=154.621 and 163.399, both P=0.000).ConclusionCombined hearing screening is of clinical significance for neonates with high-risk of hearing impairment.

Objective To research the recent efficacy of advanced cervical cancer treated with two different ways of uterine artery intervention and systemic intravenous chemotherapy respectively combined with radiotherapy.Methods Eighty-two patients with stage Ⅱ B-ⅣA cervical cancer confimed by pathology were given systemic intravenous chemotherapy combined with radiotherapy(vein group,S0 cases)and uterine artery intervention chemotherapy combined with radiotherapy(intervention group,32 cases).The therapeutic effect and remission rate of parametrium were compared.Results The total therapeutic effect rate in vein group was 90.0％(45/50),and intervention group was 93.8％(30/32),there was no significant difference between two groups(P ＞ 0.05).The remission rate of parametrium in vein group was 50.0％ (25/50),and intervention group was 75.0％(24/32),there was significant difference between two groups (P ＜ 0.05).Conclusion The effect of uterine artery intervention chemotherapy for parametrium is better than that of systemic intravenous chemotherapy.

Objective To investigate the effect of different subtypes of fractionated doses on multidrug resistance in ovarian cancer cells.Methods The human ovarian cancer cell lines SKOV3 and its drug-resistant subtype SKVCR were divided into four groups i.e., sham-irradiated, single dose (10 Gy),fractionated dose (2 Gy × 5 ) and multi-fractionated dose (1 Gy × 2 × 5).Cell sensitivity to vincristine(VCR),etoposide ( VP-16),pirarubicin (THP) and cisplatin (DDP) was measured by MTT assay.Western blot was applied to detect the expression of P-gp after irradiation.Results The doubling time of SKVCR was about 1.8-fold of that of SKOV3 cells.P-gp was expressed in SKVCR but not in SKOV3.IC50 values of SKVCR were higher than those of SKOV3.To SKOV3 cells,single dose irradiation decreased cell sensitivity to THP and DDP and fractionated irradiation decreased cell sensitivity to VCR,THP and VP-16.Multi-fractionated irradiation decreased cell sensitivity to VP-16.In SKVCR cells,all these irradiation treatments increased cell sensitivity to VCR and VP-16 but not to DDP.In addition,single and fractionated irradiation decreased P-gp expression in SKVCR cells.Conclusions Single,fractionated and multi-fractionated radiation induced chemotherapy resistance in SKOV3 cells,while reversed drug resistance to VCR and VP-16 in SKVCR cells.