The status quo and future prospects of the next generation sequencing technologies in clinical diagnostics / 中华检验医学杂志

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.

Molecular diagnosis in China：present status and challenges ahead / 中华检验医学杂志

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.

Knowledge, attitudes and practices of preconception and prenatal care in Tianjin obstetrics and gynecology personnels / 天津医药

Objective To better understand the knowledge, attitudes, practice (KAP) and relative demand of precon?ception and prenatal care (PAPC) in medical staffs of obstetrics and gynecology department in Tianjin. Methods Using ran?dom, stratified and clustered sampling approach, we selected 382 obstetrics and gynecology personnels in Tianjin. A self-ad?ministered questionnaire on PAPC KAP was employed to survey their general characteristics, their awareness of PAPC knowledge, their attitudes to PAPC, their service capabilities of PAPC and their request of continuing education. Results The respondents were lacking of KAP in Calcium and Iron supplementation, alternative preconception examination and emerging prenatal examination but present good KAP in folic acid supplemen, necessary prenatal items and classic prenatal examination. Doctors′average knowledge score was higher than nurses′. The scores in primary health care institutions was significantly higher than that in tertiary and secondary medical institutions. Young and junior medical respondents showed higher awareness than older and senior ones. Most (91.21%) respondents believe that PAPC is necessary. But 86.54%of re?spondents are not confident enough to provide comprehensive PAPC services. There were 63.74%, 54.67%, 38.74% and 32.42%of the respondents who could provide special PAPC advisory of adverse pregnancy history, thyroid dysfunction, re?productive infections and diabetes respectively. The most urgent training contents include PAPC knowledge in presence of complications or risk factors, advances in prenatal diagnosis, nutrition during pregnancy and consulting skills. Conclusion It′s necessary to intervent obstetrics and gynecology personnels to improve their cognitive level to PAPC, as well as to amelio?rate relative attitudes and behavior so they can provide more scientific and comprehensive PAPC service.

Knowledge, attitudes and practices of combined oral contraceptives in Tianjin obstetrics and gynecology personnels / 天津医药

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.

Feasibility study of ECG-triggered non-contrast-enhanced magnetic resonance venography of lower extremities / 中华全科医师杂志

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.

Genotyping of K-ras exon 2 codons 12 and 13 mutations in colorectal cancer by pyrosequencing / 中华检验医学杂志

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.

Experimental study of cryoablation on rabbit's VX2 brain tumor under MRI guidance and real-time monitoring / 中华放射学杂志

Objective To verify the feasibility of eryoablation on rabbit brain tumor under interventional MRI guidance.Methods The procedures were performed under general anaesthesia.Twenty-two New Zealand white rabbit'S brains were inoculated with VX,tumor tissue through cranial holes. MR-guided cryoablation were conducted as preliminary experiment when diameter of brain tumor grew up to 0.8 cm in size.Cryoablation lasted two cycles of 5 minutes freezing, in 2 rabbits.and of 10 minutes freezing in another 2 rabbits.After procedure and MR scanning.rabbits were executed.Specimens of rabbit'S brain showed complete necrosis pathologically after cryoablation of cycles of 10 minutes freezing.The optimal parameters of eryoablation were defined.Except for the two rabbits that died of infection,the other 16 rabbits were divided into control group and therapeutic group.According to the optimal parameters of cryoablation,cryoablation of 10 minutes freezing were performed for rabbit brain tumors in therapeutic group.Venous injeetion of 20%mannitol and subcutaneous injection of dexamethasone were used for dehydration.None of rabbit brain tumors in the control group were treated.After cryoablation,MRI scanning Was performed immediately,7 days later,14 days later and 60 days later,to observe the ablative area and the pathologic changes.The survival time wag recorded.Resuits Brain tumors could be found in all New Zealand white rabbits after inoculation except two died of inflammation.The maximum diameter of brain tumors Was from 0.7 to 1.0 elEL The tumors showed slight low signal on T1WI and slight high signal on T2WI as well as comrast enhancement after injection of Gd-DTPA.Pathologically coagulative necrosis were observed after cryoablation immediately,7 days and 14 days later,and liquefactive necrosis was found 60 days later.The correlation between MRI and pathologic appearance of tmnors are consistent.The therapeutic area ranged from 1.0 to 1.5 em.,The maximium diameter of ice ball ranged from 1.5 to 2.0 cm Conclusion CiToablation for brain tumor iS a safe and feasible therapy.

Inhibitory effects of tumor associated mitochondrial protein 12 on HepG2 cell apoptosis / 中国病理生理杂志

AIM:To explore the inhibitory effects of tumor associated mitochondrial protein 12 (TAMP12) on tumor cell apoptosis. METHODS: (1) A retrovirus expression vector was recombinated and transfected into the packaging cell line PA317. The virus particles were obtained to infect the target cell line HepG2 low expressing of TAMP12. The expression of TAMP12 mRNA was detected by RT-PCR. The subcellular localization and quantification of TAMP12 protein labeled with double fluorescein were observed under confocal laser scanning microscope (CLSM). (2) Hoechst33258 staining and flow cytometry (FACS) were used to analysis the apoptosis of HepG2 cells treated with 5-fluorouracil (5-FU). RESULTS: (1) The CLSM observation showed that TAMP12 protein was mainly expressed in mitochondria of HepG2 cells. The expressions of TAMP12 gene and protein were stable and high in transfected HepG2 cells. (2) Upon treatment with 5-FU, the transfected HepG2 cells showed a fairly integrated nucelus while the control HepG2 cells exhibited chromatin condensation, marginalization and karyorhexix. Moreover, the apoptosis rate of transfeced HepG2 cells was significantly lower than that in control HepG2 cells (P