Objective To investigate the differential expression of microRNAs(miRNAs) in transplanted kidney undergoing chronic rejection by the technique of RNA microarray.Methods Four biopsy specimens from transplanted kidney undergoing chronic rejection were harvested as test group(CR),and 3 biopsy specimens were obtained from normal renal cortex as normal control group(NC).Total RNA of each sample was extracted using Trizol reagent.miRNAs were isolated and differential expression of miRNAs were screened by miRNA array analysis.The results of miRNA array were validated by RT-PCR.The quantity and quality of all the RNA samples were checked by gel electrophoresis and absorbance at A260/280,respectively.Results It was confirmed that the isolated RNA was of appropriate quality.The results of miRNA array analysis showed that there were 63 differential expression miRNAs in CR group,of which 35 were up-regulated and 28 down-regulated.There were 9 differential expression miRNAs which distributed in 3 gene clusters: 14q32.31,22q11.21 and xq27.3.The miRNAs hsa-miR-637,hsa-miR-648 and hsa-miR-516-5p were randomly selected for relative quantification by real-time PCR.It was showed that the expression ratios of hsa-miR-637,hsa-miR-648 and hsa-miR-516-5p in AR/NC,when detected by RT-PCR,were 0.034,2.670 and 7.846,while the ratios were 0.035,2.660 and 7.857 when analyzed by miRNAs array.The results from two methods were not significantly different,so the method of miRNAs array was reliable.Conclusions It is notable that the differential expression of miRNAs existed in the transplanted kidney undergoing chronic rejection.miRNAs might be helpful in protecting the patients undergoing kidney transplantation against chronic rejection.

Objective To explore microRNA (miRNA) expression patients with β-thalassemia major.Methods MiRNA differential expression were detected in β-thalassemia major by miRNA microarray analysis and quantitative real-time PCR.Results The results of miRNA array showed that 26 differential expression miRNAs were up regulated and 30 differential expression miRNAs were down regulated.Hsa-miR-618 which expression was up regulated and hsa-miR-103a-2-5p which expression was down regulated were selected for quantitative real-time PCR detection.It was showed that the expression tendency of hsa-miR-618 and hsa-miR-103a-2-5p were consistent.So the method of miRNAs array was reliable.The target genes of 17 miRNAs which were up regulated and 24 miRNAs which were down regulated were predicted by using database software.Conclusion It is notable that the differential expression of miRNAs in patients with β-thalassemia major.It will be afforded new direction and thinking for the mechanism research and disease treatment through the further research of the miRNAs regulation pathway in β-thalassemia major.

Objective To prepare and screen out monoclonal antibodies against the receptor bind-ing domain (RBD) of Middle East respiratory syndrome coronavirus ( MERS-CoV) spike ( S) protein in mice. Methods The RBD of MERS-CoV S protein expressed in the insect-baculovirus system was purified and then used to immunize the female BALB/ c mice. The spleen cells collected from the mice were fused with myeloma Sp2 / 0 cells. The positive hybridoma cells were obtained by using limited dilution method. Enzyme-linked immunosorbent assay ( ELISA), Western blot assay and neutralization test based on the MERS-CoV pseudovirus were performed for further screening and identification. Results Twelve strains of hybridoma cells that produced the monoclonal antibodies against RBD of MERS-CoV S protein were screened out. All of the 12 monoclonal antibodies (McAbs) could have specific reaction with the RBD of MERS-CoV S protein as indicated by the results of ELISA. Of the 12 McAbs, two were identified as the immunoglobulin M (IgM) isotype and the rest were IgG1 isotype by using double antibodies sandwich ELISA. Four McAbs including 1F1, 2E4, 3C3 and 3E6 were identified as having neutralizing activity by the neutralization test based on MERS-CoV pseudovirus. Results of the Western blot assay showed that the four McAbs (1F1, 2E4, 3C3 and 3E6) could have specific reaction with the RBD of MERS-CoV S protein, but no cross-reac-tion with that of SARS-CoV S protein. Conclusion Twelve mouse-derived McAbs against the RBD of MERS-CoV S protein were obtained. The prepared hybridoma cells showed the characteristics of high speci-ficity and stability in antibody secretion. Four out of the 12 McAbs were proved to have neutralizing activity.

Objective To investigate healthy status and disease distribution of couples through analysing the results of pre‐preg‐nancy laboratory examination and to explore high risk factors ,in order to provide scientific references for the realization of eugenics . Methods The routine blood test ,serum markers of hepatitis B ,five items(TORCH) examination ,detection of female vaginal secre‐tions were carried out in 8 313 couples planning pregnancy from 2013 to 2014 ,and the results of pre‐pregnancy health examination were analysed and evaluated .Results The thyroid dysfunction accounted for 17 .43% of all women undergoing examination ,which was the most common disease .Among items carried out in all couples ,thalassemia and positive of hepatitis B surface antigen were the most common ,accounted for 10 .20% and 9 .94% respectively .Conclusion The pre‐pregnancy health examination could screen high risk factors ,which is benefit for guiding eugenics and effectively reducing the birth defects and adverse pregnancy and is a sig‐nificant measure for eugenics .

Objective To compare the efficacy of surgically managing stress urinary incontinence (SUI) with tension free vaginal tape(TVT)and autologous fascia lata pubovaginal sling(Lata) Methods Sixty seven patients suffering from SUI entered this prospective study The patients were divided into two groups randomly, 48 in TVT group and 19 in Lata group The average follow up time post operation was 20 months in TVT group and 37 months in Lata group Results The operating time was(27?5)min in TVT and (125?13)min in Lata The volume of blood loss during surgery was:(27?6)ml in TVT and (67?11)ml in Lata The post operative recovery period in the hospital was: TVT, 1 8 days and Lata, 7 2 days The differences between the two groups for these three parameters were statistically significant( P

Objective To investigate methods for prenatal molecular diagnosis of fetuses at high risk for X-linked adrenoleukodystrophy(X-ALD).Methods The amniotic fluid was obtained and genomic DNA was isolated from amniotic fluid cells.Maternal contamination was evaluated by paternity test.PCRRFLP,sequencing and denaturing high performance liquid chromatography(DHPLC)were used to detect the ABCD1 gene of fetal genome.Results In the pedigree 1,the PCR product(799 bp)of the fetus 1 and her father(normal control)could be digested with BcnI. No P560L mutation,which was present in the index patient,was detected in the ABCD1 gene from the genomic DNA of the fetus 1 using direct sequencing.In the pedigree 2,the PCR product(232 bp)of the fetus 2 and her father could not be digested with MaeI and no Q177X mutation,which was present in the propositus,was detected in the ABCD1 gene from the genomic DNA of the fetus 2 using direct sequencing.In the pedigree 3,the PCR product(271 bp)was digested with AciI.the pattern of the fetus 3 and the propositus being the same,and the R617C mutation was found in the ABCD1 gene from the genomic DNA of the fetus 3 using direct sequencing.In the pedigree 4,the PCR product(269 bp)was analyzed with the DHPLC,and the pattern of elution peaks of the fetus 4 and her father was similar,but different from that of the propositus.No K276E mutation was detectable in the ABCD1 gene from the genomic DNA of the fetus 4 by using direct sequencing.Judging from the sex of the fetuses,fetuses 1 and 2 were normal homozygotes,fetus 3 was an ALD hemizygote,and fetus 4 was a normal hemizygote.Conclusion A new protocol for X-ALD prenatal molecular diagnosis is proposed,which would ensure the accuracy of prenatal diagnosis.

Objective To analysis of the detection result of amniotic fluid chromosome which in NIPT high-risk pregnant women.Methods Amniotic fluid cells via amniotic cavity puncture were cultured and analyzed,the chromosome karyotypes were observed.Results The highest positive predictive value of NIPT was for trisomy 21(85.00%),then trisomy 18(75.00%),sex chromosome abnormalities(68.00%),other chromosome abnormalities(41.67%),trisomy 13 (25.00%).Conclusion The highest accuracy of NIPT was shown in detection of Down''s syndrome by NIPT.NIPT was screening test which is effective and noninvasive in prenatal diagnosis.Amniotic fluid Chromosomal karyotype analysis was the gold standard in the diagnosis of fetal chromosomal disease.

Objective To investigate the effects of cochlear implantation on residual hearing and to evaluate the potential impact of long -term electrical stimulations on residual hearing .Methods 58 hearing impaired children with cochlear implants were included in this study .All subjects could cooperate with behavioral audiometry .Audio-metric evaluations were carried out pre -implantation and 3 ,12 ,24 months post -implantation respectively .Of 58 subjects ,43 were followed up more than 1 year and 17 were followed up more than 2 years .Results All 58 subjects showed significant differences (P<0 .05) between pre- and 3 months post-implantation of residual hearing at the individual frequencies of 0 .25 ,0 .5 ,1 ,2 and 4 kHz .43 subjects followed up more than 1 year showed statistic differences (P<0 .05) between pre- and 3 months post -implantation ,pre- and 12 months post-implantation at 0 .25 ,0 .5 ,1 ,2 and 4 kHz respectively .Comparing 3 months with 12 months post -implantation ,there were sta-tistic differences at 0 .25 and 0 .5 kHz ,while no significant difference (P>0 .05) at 1 ,2 and 4 kHz .Of 17 subjects followed up more than 2 years ,there were significant differences (P<0 .05) between pre- and various return visits post-implantation .Post-implantation return visits ,there were significant differences between 3 months and 12 , 24 months at 0 .25 and 0 .5 kHz respectively ,not any significant differences on 1 ,2 and 4 kHz .There were no sig-nificant differences on each frequency between 12 months and 24 months post- implantation .Conclusion Residual hearing after cochlear implantation could decrease to some extent for various reasons .There were significant differ-ences between 3 and 12 months post-implantation at 0 .25 and 0 .5 kHz .Not any significant differences were ob-served between 12 months and 24 months post-implantation at each frequency .

Objective To explore the effect of health belief model on compliance behavior of patients with PICC(peripherally inserted central catheter).Methods According to admission order,75 cases of PICC were divided into control group(n=36)and observation group(n=39).Patients in the control group received conventional health education and regular visits after discharge and besides the regular methods,patients in the observation group were intervened by health belief model for the compliance behavior.All patients of two groups were investigated with self-designed scale for compliance behavior.Result The compliance behavior of the observation group was better than that of control group with significant difference(P<0.05).Conclusion The health belief model of education can improve the compliance behavior of PICC patients after discharge from hospital.

Objective To compare the prenatal ultrasonic characteristics between left and right isomerism as referring to autopsy outcomes.Methods Between November 2007 and July 2010,fetuses with isomerism that were confirmed by autopsy,were identified from 1200 prenatal ultrasonic scans.Their abnormal spectrums and ultrasonic features were analyzed and comparied between left and right isomerism.Results A total of 18 fetuses with right isomerism and 4 fetuses with left isomerism were detected and confirmed.The major findings of the right isomerism were;viscerocardiac heterotaxy (17,94%),anomalous pulmonary venous connection (16,89%),complete atrioventricular septal defect (15,83%),juxtaposition of the descending aorta and inferior vena cava(12,67%),left persistent superior vena cava (11,61%),right aortic arch (10,56%) and univentricle (10,56%).As for the left isomerism,all had viscerocardiac heterotaxy and interruption of inferior vena cave,three of them had heart block,complete atrioventricular septal defect and hypoplasia of aorta,two of them had univentricle.Conclusions Viscerocardiac heterotaxy,complete atrioventricular septal defect,univentricle are the most common anomalies detected.Right isomerism cases usually exist with anomalous pulmonary venous connection,juxtaposition of the descending aorta and inferior vena cava as well as right aortic arch.The left isomerism cases usually have interruption of inferior vena cave,heart block and hypoplasia of aorta.