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Objective To establish a rapid detection method for zika virus based on direct amplification re-al-time fluorescent quantitative reverse transcription polymerase chain reaction(RT-PCR)technique.Methods A direct amplification RT-PCR technique for the rapid detection of zika virus in 5 samples(whole blood,serum,saliva,throat swab and urine)was established by using a special function DNA polymerase and a preferred PCR enhancer.Results The detection limits of the 5 samples were 103 PFU/mL in serum,102 PFU/mL in urine,throat swab,and saliva,and 104 PFU/mL in whole blood.The coefficient of goodness-fit of stand-ard curves was above 0.98,and the amplification efficiency was 90％-110％.Zika virus nucleic acid was suc-cessfully amplified,but non-zika virus nucleic acid was not amplified.Based on the repeatable detection of sam-ples from urine,whole blood,and saliva,the variation coefficient of 6 repeated Ct values at 106 PFU/mL and 102 PFU/mL concentrations were all＜5％.The zika virus detection method established by the direct amplifi-cation RT-PCR technique was consistent with the detection results of conventional RT-PCR technique.Only two serum samples were detected in eight zika virus samples,and the remaining 62 non-zika virus samples and 12 negative samples were not amplified.Conclusion A rapid detection method for zika virus based on direct ampli-fication RT-PCR technique is successfully established.The method is simple,rapid,sensitive and specific.
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Objective:To investigate the correlation between the promoter methylation level of Dickkopf-related protein 1 (DKK1) gene and diabetic microangiopaopathy complicated with osteoporosis.Methods:Patients with type 2 diabetes mellitus (T2DM) microangiopathopathy who were admitted to our hospital from Jan. 2019 to Dec. 2022 were collected as research objects, and divided into observation group (44 cases) and control group (58 cases) according to whether they were complicated with osteoporosis. Bone mineral density (BMD) of lumbar spine (L1-4) was measured, and bone metabolism indexes, including serum calcium, serum phosphorus, 25-hydroxy vitamin D3[25-hydroxy vitamin D3, 25- (OH) D3], PTH, C-terminal telopeptide of typeI collagen (CTX), procollagen of aminoterminal propeptide (PINP) and tartrate resistant acid phosphatase (TRACP) levels were detected; The promoter methylation level of DKK1 gene was determined.Results:The methylation level of DKK1 gene promoter in the observation group was 5.17%±0.73%, which was significantly higher than that in the control group (3.81%±0.61%), with statistical significance ( t=5.22, P<0.001). The 25- (OH) D3 level, PTH and lumbar bone density in the observation group were significantly lower than those in the control group, while the CTX and TRACP levels were significantly higher than those in the control group ( t was 5.58, 4.35, 4.12, 4.05 and 4.17, respectively, P<0.001). In all patients, the promoter methylation level of DKK1 gene was significantly positively correlated with CTX and TRACP ( r was 0.41 and 0.39, P was 0.006 and 0.027, respectively), and significantly negatively correlated with PTH and lumbar bone density ( r was -0.38 and -0.43, respectively). P=0.015 and 0.003, respectively). ROC curve analysis showed that the area under the curve of DKK1 methylation level to distinguish type 2 diabetes microangionopathy with and without osteoporosis was 0.841 (0.762-0.921), and the sensitivity and specificity were 86.4% and 72.4%, respectively. Conclusion:The methylation level of DKK1 gene promoter is associated with osteoporosis and bone metabolism in T2DM patients with microangiopathia.
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Osteoarthritis is a kind of joint disease with cartilage injury as the main pathological changes.There is no effective treatment for them.With the discovery,research and application of microRNA (miR),it also provides new hope for the treatment of osteoarthropathy.At present,the role of miR in the pathogenesis of cartilage injury has been fully recognized.In this paper,we reviewed the role of miR-140 in the development of osteoarthritis and its important target genes in recent years.
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Osteoarthrosis is a disease characterized by degeneration of articular cartilage,inflammation of the synovial membrane in the joint,and changes of the surrounding joints and subchondral bone.The main pathologic changes of osteoarthrosis are chondrocytes reduction and extracellular matrix degradation,which then affect the synovial membrane and articular cartilage bone plate accompanied by osteophyte formation,lead to a series of joint symptoms and signs.Modern medical research shows that long non-coding RNA (lncRNA) plays a certain role in the pathogenesis and development of osteoarthrosis.In this paper,the advances in the study of lncRNA and osteoarthrosis are reviewed.
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Objective To investigate the effect of telmisartan on the expression of APN,hs-CRP,TNF-α and the prognosis of hypertensive patients with unstable angina pectoris. Methods Totally 100 hypertensive patients with unstable angina pectoris were randomly divided into the telmisartan group(n =50) and the losartan group(n =50),each group with oral administration. The blood pressure controlled condition, APN, hs-CRP and TNF-α content changes and the occurrence of cardiovascular accident after treatment were observed in both two groups. Results In both groups, blood pressure returned to normal in 8 weeks,with no significant difference(P ＞ 0.05). After treatment,APN level in the telmisartan group increased faster than that in the losartan group(P ＜0.05) ,while the hs-CRP,TNF-αdecreased significantly(P ＜ 0.05). After 8 weeks, the occurrence of cardiovascular accident in the telmisartan and losartan groups was 10.0% (5/50) and 24. 5 % (12/49) respectively in the following 4 months (P ＜ 0.05). Conclusion Telmisartan has significant effect in regulating APN, hs-CRP, TNF-α expression in hypertensive patients with unstable angina pectoris, which is able to decrease the short-term occurrence of cardiovascular accident.
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Objective To study the clinical diagnostic value of transvaginal ultrasonic in early cervical carcinema and cervical neoplasis.Methotis The suspected patients with cervical carcinoma and cervical neoplasis were detected with transvaginal ultrasonography,liquid based cytology and cervical biopsy.The sonograms of transvaginal ultrasonic were retrospectively analyzed.Results In early cervical carcinoma and cervical neoplasis,the diagnostic sensitivity of transvaglnal ultrasonic was 90.9%and 83.3%;and the diagnostic specifity of transvaginal ultrasonic was 70.6% and 60.0%;the rate of missed diagnosis of transvaginal ultrasonic was 9.1% and 16.7%.Conclusion Transvaginal ultrasonic plays an important part in the clinical diagnosis of early cervical carcinoma.
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Objective To investigate 1762T、1764A double position mutation in the Basic Core Promoter(BCP) of hepatitis B virus and reveal its relation to clinical symptoms and HBeAg phenotype. Methods microplate sandwich hybridization technique was used to detect BCP double position mutation. One common capture probe and one mutant specific detector probe as well as one wild type detector probe were synthesized and hybridized with amplified HBV DNA from the sera of hepatitis B patients. The results of hybridization were exhibited with ELISA. Results 147 hepatitis B patients who had been confirmed HBV DNA positive were screened. 51 patients were BCP double position mutation, 42 of which were BCP single position mutation and, 9 were mix mutation(both mutation and wild type were positive). BCP mutant was detected in 36 of 117 with chronic hepatitis and, 8 of which were mix mutants. Moreover, BCP mutant was detected in 7 of 30 with acute hepatitis in 25 of 78 with HBeAg positive were mutant and in 26 of 65 with HBeAg negative were mutant.Conclusions (1) The rate of BCP double position mutation in chronic hepatitis B patients is higher than that in acute hepatitis B patients. (2) BCP mutation may impair HBeAg expression. (3) PCR microplate sandwich hybridization ELISA is a sensitive and efficient method for detecting gene mutations.