Molecular diagnosis is rapidly developed in recent years , mainly applicated in the diagnosis of hereditary disease , infectious pathogens, tumor susceptibility and molecular typing , companion diagnosis and prognosis assessment , playing more and more important role in many diseases diagnosis and treatment.Molecular diagnosis was developed from the eighties of the last century in our country .Nowadays, the mainly applied technologies in the clinical laboratory include fluorescence in situ hybridization , quantitative PCR, microarray and DNA sequencing. These molecular technologies make up for the insufficiency of routine testing and take up a central role in the development of modern laboratory medicine . With the continuous development in transformation research of molecular technology recent years , there will be more molecular diagnostic techniques applied in clinicaldiagnosis in the future .But it still exists some drawbacks in the performance of molecular diagnosis in our country according to the current situation , such as imbalanced regional development , mismatched policies, non-standardized laboratory construction , deficiency of quality control and supervision , etc., which requires the joint effort of the government , hospital, professional association and clinical laboratory itself to promote the healthy and orderly development of molecular diagnosis.

During the past decade, tremendous progress has been made in terms of speed, read length, and throughput, along with a sharp reduction in per-base cost.Together, these advances democratized next generation sequence (NGS) and paved the way for the development of a large number of novel NGS applications in clinical diagnostics, especially in the field of non-invasive prenatal detection, rare genetic disease and cancer companion diagnostics.As technology advances, long-read single molecule sequencing began to emerge.Single cell, long-reads, transcriptome, and low cost will be the NGS direction.Due to the special nature of clinical testing, the current NGS clinical application system,including genetic counseling, testing standards, quality control, supervision, database construction etc, does not match the national conditions well and still faces a few challenges, needs to be constantly improved through the routine clinical practice in the future.

Objective To develop a canine model of neurogenic bladder and to evaluate it with urodynamics. Methods Eight female dogs were divided into 2 groups:4 in suprasacral and 4 in infrasacral group.The spinal cord was transected at the level of 5th～6th lumbar intervertebra foramen,and sacral cord was kept intact in suprasacral model while in infrasacral model it was destroyed completely.The urodynamic studies were performed in each dog at the time points of preopertation and 6 months postoperation in both groups.The parameters included bladder capacity,detrusor pressure,bladder compliance and urethral pressure. Results The dogs after operation could never move their tails and keep their hips dry.Their hind limbs presented flaccid paralyses and the abdomens dilated during the first postoperative week.Then the mobility of the hind limbs in both groups recovered slowly,but the abdominal signs were improved obviously only in suprasacral dogs.In suprasacral group,bladder capacity and compliance were decreased by 37.3% and 52.1%,respectively, but detrusor pressure (33.3%) and urethral pressure (17.3%) were significantly increased( P

Objective To evaluate catheterization in pancreatic duct before endoscopic papillary bal-loon dilation (EPBD)to prevent pancreatitis after EPBD.Methods Forty-three patients with normal serum amylase levels,diagnosed as having bile duct stones,underwent EPBD.Twenty-three were assigned to experi-mental group,where catheters(ERCP imaging tube)were placed in pancreatic duct before EPBD,then the pa-pillary balloon was expanded to 10 mm.Twenty were assigned to control group where eight-millimeter-diameter papillary balloon was used to remove the stones.The serum amylase levels before EPBD,6 hours and 24 hours after EPBD,the incidence of pancreatitis and high serum amylase levels associated with EPBD,as well as the mean time and success rate of removing the stones of the two groups were compared.Results Post-EPBD pan-creatitis occurred in one patient in experimental group (4.35%),and seven in control group (35.00%), which was significantly different(P <0.05).Meanwhile,the mean levels of serum amylase 6 h and 24 h after EPBD in the experimental group were (102.61 ±98.99)U /L and (60.35 ±26.18)U /L respectively,lower than those in the control group (398.25 ±259.32)U /L and (230.50 ±281.31)U /L(P <0.05).After the papillary balloon was expanded to 10 mm in experimental group,the mean time of removing stones was (10.43 ±2.27)min,which was shorter than that of control group (17.90 ±4.49)min (P <0.05).Stone-re-moving rate of two groups had no difference and they all succeeded one time.Conclusion Placing catheter in pancreatic duct before EPBD to prevent pancreatitis after EPBD makes it easier to remove stones in shorter op-eration time.It can prevent pancreatitis and high amylase blood disease after EPBD.

Objective To understand the knowledge, attitude, practice (KAP) status and relative demand to combined oral contraceptives (COCs) in medical staffs of obstetric and gynecological department in Tianjin, providing reference basis for the effective intervention measures. Methods By using random, stratified and clustered sampling approach, we selected 382 medical staffs of obstetric and gynecological department in Tianjin. A self-administered Questionnaire on COCs KAP was employed to survey the general characteristics, awareness of COCs knowledge, attitudes to COCs, service capabilities of contraceptive counseling, prescribing behavior and request of continuing education. Results The respondents showed de?fects in COCs indications, non-contraceptive benefits, relationship with cancer and birth defects. The average knowledge score was higher in doctors than that of nurses. The higher the titles and qualifications, the higher the knowledge score. A to?tal of 90.11%of respondents believed that strengthening the contraceptive guidance can help to reduce pregnancy. Strength?ening the interpretation of misunderstanding (81.04%) and enhancing the treatment and follow-up of adverse reactions (62.09%) can help to reduce stop taking. Only 25.82%of respondents could provide detailed COCs relative consulting servic?es;42.96%of obstetricians and gynecologists refused prescribing COCs for women over the age of 40 for the purpose of con?traception. A total of 93.13%medical personnel showed demand on relative training. Conclusion It’s necessary to perform continuing education to medical staffs via different intervention, so as to improve their cognitive level to COCs, ameliorate COCs relative attitudes and behavior, and help them providing better service to women of childbearing age.

ObjectiveTo investigate the clinical significance of pyrosequencing assay for determining K-ras mutations in exon 2 codons 12 and 13 in clinical colorectal cancer tissues.Methods Genomic DNA,extracted from K-ras mutant cell lines SW480 (homozygous,c.35G ＞ T), DLD-1 (heterozygous,c.38G ＞ A) and wild-type HT-29,was first used as the sequencing template respectively to test the accuracy of pyrosequencing methodology.The SW480 and DLD-1 DNA was separately mixed with wild-type HT-29 DNA in proportions of 2％,3％,5％,10％,20％,30％ and 50％,the sensitivity for mutation detection was measured separately by pyrosequencing assay and directed Sanger DNA sequencing in the serial DNA mixture samples.The pyrosequencing assay results were compared with the corresponding Sanger sequencing and the datas were analysized by Fisher exact test.Pyrosequencing analysis was then performed for screening K-ras exon 2 mutations at codons 12 and 13 on DNA isolated from a panel of 30 colorectal cancer samples derived fromclinicalformalin-fixed andparaffinembedded(FFPE)tissues.ResultsCancer cell lines with known K-ras mutations ( SW480 and DLD-1 ) were readily detectable by pyrosequencing-based analysis.When the proportions of mutant colorectal cancer cell line DNA were 5％ and 10％ content,the mutation rates of K-ras gene detected by conventional Sanger DNA sequencing were 33.3％ (4/12) and 58.3％ (7/12) respectively,whereas the mutation rates detected by pyrosequencingbased assay were 91.7％ (11/12) and 100％(12/12) respectively,there were significant differences between those two sequencing methodology ( P ＜0.05).Furthermore,we found 10 patients with K-ras exon 2 point mutations at codons 12 and 13 by pyrosequencing-based assay from 30 colorectal cancer FFPE tissues,the point mutation rate was 33.3％ (10/30) and all of the mutations determined were heterozygous.The codon 12 was most frequently affected [30％ (9/30)].Mutations with the highest frequency were G ＞ A transitions [ 50％ ( 5/10 ) ],followed by G ＞ T transversions [ 30％ ( 3/10 ) ].Conclusion The pyrosequencing assay provides an accurate and sensitive method for mutation screening of K-ras exon 2 codons 12 and 13 in routine diagnostic specimens,thereby allowing the selection of the cancer treatment in clinical individualized practice.

Twelve healthy volunteers underwent ECG-triggered non-contrast-enhanced magnetic resonance venography(NCE-MRV)of lower extremities on a 1.5T MR scanner.Image quality was assessed quantitatively and qualitatively with a 17-segment model.ECG-triggered (NCE-MRV) was successfully performed in all patients.Mean signal to noise ratio and contrast to noise ratio of lower extremities were 66 ± 23 and 53 ± 22 respectively.Excellent and good image qualities were achieved in 62％ and 31％ of all segments respectively without non-evaluable segment.Thus ECG-triggered NCE-MRV of lower extremities is feasible with reliable image quality.

Objective To grasp the epidemic features of Kaschin-Beck disease in different ecotypic areas in Gansu Province,in order to provide ecological basis for Kashin-Beck disease control.Methods Totally 37 counties with Kashin-Beck disease were divided into four ecological areas,villages with historical serious condition in township and townships with historical serious condition in county were investigated.Clinical examination and Xray of right hand of all 7-12-year-old children in the villages were carried out,while investigation of prevention and control measures was performed.Results Among 27 966 children from four ecological areas,the clinical detection rate was 0.05％ (14/27 966) and the X-ray positive rate was 1.26％ (353/27 966),metaphyseal rate was 1.25％ (350/27 966),bone-side positive rate was 0.01％ (3/27 966) and no case of epiphyseal and carpal were discovered.The clinical detection rate in children in the four ecological areas was not statistically significant (x2() =7.757,P ＞0.05),the Loess Plateau-gully region of Longdong [0.09％ (10/11 604)] ＞ the Anyon area of Qinling of Longnan [0.04％ (3/7 969)] ＞ Alpine grassland meadow area of Gannan [0.02％ (1/4 021)] ＞ the Loess Plateau-hills region of Longzhong [0 (0/4 372)].The detected rate of X-ray in four ecological areas was statistically significant (x2 =18.133,P ＜ 0.05),the Anyon area of Qinling of Longnan [1.49％ (119/7 969)] ＞ the Loess Plateau-gully region of Longdong [1.41％ (164/11 604)] ＞ the Loess Plateau-hills region of Longzhong [0.89％ (39/4 372)] ＞ Alpine grassland meadow area of Gannan [0.77％ (31/4 021)].Comprehensive prevention and control measures on Kaschin-Beck disease were different in different ecotypic areas.Relocation,long-distance education and selenium supplement measures had not been implemented in the four ecological areas.Conclusion The epidemic situation of Kaschin-Beck disease is different in different ecotypic areas in Gansu Province,which maybe related to ecological environment,but is under a state of control.

Objective To evaluate the accuracy and feasibility of time-resolved immunofluorometric assay (TRI FA) for detection of HBsAg based on Abbott automated chemiluminescence immunoassay(CMIA),so as to carry out this project in primary hospitals,and provide reference for individual antiviral strategy and prediction of therapeutic effect.Methods Serum of 157 patients infected with hepatitis B virus were detected with CMIA and TRIFA,specimens with HBsAg titers exceeding the detection limit were firstly diluted,then performed quantitative analysis.HBsAg levels were divided into 4 groups:≤100 IU/mL,101-1 000 IU/mL,1 001-20 000 IU/mL,and ＞ 20 000 IU/mL,quantitative correlation between two methods was analyzed.Results The linear regression equation of two methods was Y=2.323X-896.3,correlation coefficent r=0.943,P＜0.001.CMIA was as a reference,4 groups were divided for analysis,results showed that when detected specimens was at low concentration of HBsAg,TRIFA value was low compared with CMIA method,while detected specimens was at high concentration of HB sAg,CMIA value was high,two reagents had good consistency in the detection of different concentrations of HBsAg(both P＜0.05),when concentration was at 1 001-20 000 IU/mL,consistency was the best.Conclusion The accuracy of two reagents in the quantitative detection of HBsAg is similar,and the best correlation of detection value is 1 000-20 000 IU/mL.TRIFA assay has wide application for its low-cost and easy to be operated,which is especially suitable for primary hospitals.

Objective To observe the condition of Keshan disease (KSD) in Gansu Province in order to provide a scientific basis for prevention and control of the disease. Methods In 2011, according to “The national Keshan Disease Monitoring Program (trial edition)”, based on searching of KSD cases, 12 villages of 12 towns in 6 counties of Gansu Province, were selected as surveillance sites. All the residents in surveillance sites were clinically examined and given 12-lead ECG tracings; suspected cases were taken anterior chest X-rays in the distance of 2 meters, while staple food and life conditions of the residents were surveyed. Diagnosis of KSD was based on“Keshan Disease Diagnostic Criteria”(WS/T 210-2011). Results Among investigated 5 484 residents, the incidence of abnormal electrocardiogram was 15.41%. Of the 196 X-ray films, 61 cases had enlarged heart(in which 22 of mild, 21 of moderate and 18 of severe). Two hundred and forty-three cases of KSD were detected, the general detection rate was 4.43%, in which 31 cases of chronic, the detection rate was 0.56%; 212 cases of potential, the detection rate was 3.87%. In all cases, 47 cases were under the age of 30, including 46 cases of potential and 1 case of chronic. The major abnormal electrocardiogram change of KSD cases was ST-T changes[22.68%(71/313)], followed by complete right bundle branch block[16.29%(51/313)], low voltage[12.46%(39/313)], and left anterior fascicular block[6.71%(21/313)]. Per-capita annual income in surveillance site was 1 763 Yuan;and the major staple food was flour, accounted for 87%, and the staple food rice accounted for 5%. Conclusion The detection rate of KSD in Gansu Province is mainly to potential, and there is a cosiderable portion of patients under the age of 30;monitoring and investigation should also be strengthened in the younger age of KSD cases detected village.