1.Detection and Identification of Amomum Viosum by Fluorescent Quantitative PCR
China Pharmacist 2015;(6):946-948
Objective:To analyze the genetic sequence characteristics of amomum viosum and establish a rapid identification meth-od for amomum viosum by fluorescent quantitative PCR based on DNA analysis. Methods:Amomum viosum and the other samples be-longing to the same genera were collected and identified by experts in the domain. DNA was isolated using commercial kits. The prim-ers and probe were designed according to the conserved region of ITS in amomum viosum. The reaction conditions were optimized to es-tablish the fluorescent quantitative PCR method for the rapid detection of amomum viosum. Results:The fluorescent quantitative PCR method for the rapid detection of amomum viosum was set up. The method could identify amomum viosum successfully, while those samples in the same genera were without amplification curves. Conclusion: Amomum viosum can be identified rapidly by fluorescent quantitative PCR besides the traditional identification by experts.
2.Application of critical-care pain observation tool for the evaluation of pain in elderly patients with mechanical ventilation
Meng CHUN ; Zhang YAN ; Liang YU
Chinese Journal of Geriatrics 2011;30(12):1008-1011
ObjectiveTo assess the degree of pain in elderly patients with mechanical ventilation in ICU using critical-care pain observation tool(CPOT) and to choose the correct sedative and analgesic method.Methods 110 elderly patients in ICU after neurosurgery were divided into three assessment stages,every stage had two record points and total six points (T1-T6):the first stage (intubation and unconsciousness,T1-T2),the second stage (intubation and consciousness,T3T4 ) and the third stage(extubation and consciousness,T5-T6 ).Among them T1,T3and T5were nonnocuity assessment points of every stage,while T2,T4 and T6 were nocuity assessment points of every stage.The assessment time was one minute at every point.After recorded at every point in second and third stages,patients were asked to use the pain intensity descriptive scale (PIDS) themselves.CPOT,heart rate and mean arterial pressure (MAP) from T1 to T6 were recorded as well as PIDS from T3 to T6 in second and third stages.Results In the three stages,CPOT〔(26.8 vs.0.54,3.36 vs.1.20,2.78 vs.0.68) scores〕,HR〔(95 vs.85,94 vs.82,94 vs.84)beat/min〕 and MAP〔(95 vs.85,95 vs.87,94 vs.87)mm Hg〕 at T2,T4and T6 were higher than T1 (t=-42.89,-55.95,-55.38),T3 (t =- 5.52,- 11.33,- 11.78) and T5 ( t =- 5.54,- 9.95,- 11.33 ) ( P< 0.05 ).The PIDS at T4 and T6were higher than at T3and T5in the second and third stages 〔(2.52 vs.1.69,2.12 vs.1.44)scores〕 (P<0.05).The correlation coefficient between CPOT and PIDS at T3 and T4 in the second stage were 0.49 and 0.58,respectively (P<0.05),and between CPOT and PIDS at T5 and T6 were 0.52 and 0.59 in the third stage,respectively (P < 0.05),and they both reached moderate correlation.ConclusionsCPOT may be an effective way to assess the degree of pain in elderly patients with mechanical ventilation at present.
4.Effect of propofol on a-amino-3-hydroxy-5-methyi-4-isoxa-zolep-propionate receptors AMPA GluR1 subunit and long-term potentiation (LTP) in hippocampal slices in aged rats
Yuzheng ZHENG ; Yan ZHANG ; Yu LIANG
Chinese Journal of Geriatrics 2013;(3):330-332
Objective To observe the effect of propofol on phosphorylation of a-amino-3-hydroxy-5-methyl-4-isoxa-zolep-propionate receptors (AMPARs) GluR1 subunit and long-term potentiation (LTP) in cultured hippocampal neurons in aged rats.Methods A total of 30 18-month-old rats were decapitated,the brains were rapidly removed and hippocampal slice were prepared.The slices were randomly divided into control group (perfused with artificial cerebrospinal fluid,n=10),propofol-treated group (perfused with propofol in artificial cerebrospinal fluid,n=10)and propofol+ phorbol-12-myristate-13-acetate (PMA)-treated group (perfused with propofol and phorbol ester in artificial cerebrospinal fluid,n=10).Extracellular excitatory postsynaptic potentials (EPSP) were recorded from the CA1 region of hippocampal slices.After perfusion for 20 min,LTP was induced using higher-frequency stimulation (HFS,100Hz,400 pulse) by the Schaffer-collateral pathway.The phosphorylation of AMPA-GluR1 subunit was assayed in cultured rat neurons by Western blot.Results The value of EPSP in propofol-treated group (105.50 ± 3.77) was much lower than in control group (242.10±14.68) and in propofol+ PMA-treated group (239.40±8.98) (F=2.90,P<0.05),and there was no significant difference in the value of EPSP between control group and propofol+ PMA-treated group (P>0.05).The level of P-Glu1/Glu1in propofol-treated group (0.68±0.15) was much lower than in control group (1.67±0.20) and in propofol+PMA-treated group (1.57±0.18) (F=6.84,P<0.05),while there was no difference in the level of P-Glu1/Glu1 between control group and propofol + PMA-treated group (P > 0.05).There was no difference in the value of GluR1/β-actin among the three groups (F=0.31,P>0.05).Conclusions Propofol possesses the ability to inhibit LTP induction and attenuate AMPA receptor GluR1 subunit phosphorylation through modulation of PKC pathway.
5.Construction of DC-SIGN eukaryotic expression vector and establishment of BHK21 cell line stably transfected by the recombinant vector
Yu WANG ; Jinqi YAN ; Liang ZHANG ; Yue WANG ; Jiyun YU
Medical Journal of Chinese People's Liberation Army 1982;0(03):-
Objective To construct a eukaryotic expressing vector harboring human DC-SIGN, and establish a BHK21 cell line stably and highly expressing DC-SIGN. Methods The DC-SIGN gene fragment which contained Not I and BamH I sites was amplified by PCR from pUNO-hDCSIGN1Aa plasmid, digested with Not I and BamH I, and then cloned into an eukaryotic expression vector pIRES-neo to construct eukaryotic expression vector pIRES-neo-DC-SIGN. The recombined plasmid was identified with Not I and BamH I enzyme digestion and sequencing, the latter was then transfected to BHK21 cells by LipofectamineTM 2000. After screening culture by G418, BHK21 cell line stably expressing DC-SIGN was established. The expression of DC-SIGN was identified by flow cytometry, Western blotting and immunofluorescence method. Results The gene sequence of DC-SIGN was consistent with that of design. PCR and double enzyme digestion analysis showed that the recombinant plasmid pIRES-neo-DC-SIGN was constructed successfully. After transfection, positive clones were selected with G418. After limiting dilution assay, BKH21 cell lines stably expressing DC-SIGN were established. The detection result of flow cytometry showed that the expression ratio of DC-SIGN positive clones was close to 90%. The result of immunofluorescence displayed that the expression of DC-SIGN was mostly located on the surface of cell membrane. Western blotting displayed the specific band of DC-SIGN protein. It showed that the BHK21 cells stably expressing DC-SIGN were successfully established. Conclusion DC-SIGN eukaryotic expression vector has been successfully constructed. The successful establishment of BHK21 cell lines which can stably express DC-SIGN provides a substantial foundation for further study on the DC targeting vaccines.
6.MicroRNA-215 is a potential prognostic marker for cervical cancer.
Hua, LIANG ; Yan, LI ; Ruo-yu, LUO ; Fu-jin, SHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(2):207-12
Recently, microRNAs (miRNAs) have been shown to be involved in multiple biological pathways that can influence tumor progression and metastasis and they can serve as prognostic biomarkers in many cancers. The present study examined the prognostic significance of miR-215 in cervical cancer. The paraffin-embedded paired cervical scrape samples and tumor tissue samples from 302 patients with stage II cervical cancer were detected for the expression of miR-215 by using qRT-PCR. A miR-215-based classifier was established by using the Cox regression model. The prognostic and predictive accuracy of this classifier was determined in both the internal testing group of 138 patients, and the external independent group of 280 patients. Moreover, cervical cancer HeLa cells overexpressing miR-215 (HeLa-miR-215) were constructed and subcutaneously injected into the nude mice to examine the effect of miR-215 on tumor growth and metastasis in vivo. The results showed that the expression level of miR-215 was significantly higher in cervical cancer tissues than in paired normal tissues (P<0.0001). When patients were classified into high- and low-risk cancer progression groups according to miR-215 level, the 5-year disease-free survival in high- and low-risk groups were 43% (95% CI: 32.1-51.6) and 67% (95% CI: 48.6-77.3) (hazard ratio [HR] 2.02, 95% CI: 1.16-3.52; P=0.013) respectively. Moreover, the expression level of miR-215 was negatively associated with survival rate in patients at TNM stage T3 (HR: 3.317; 95% CI: 1.18-5.14, P=0.017) and TNM stage T4 (HR: 3.48; 95% CI: 1.49-4.45, P=0.008). Tumor volume in nude mice injected with HeLa-miR-215 cells was significantly larger than that in mice injected with control HeLa cells. It was concluded that the expression level of miR-215 is associated with cervical tumor progression and worse survival rate, suggesting that it may serve as a potential prognostic marker to identify patients at higher risk of recurrence.
7.Quantitative Analysis of Contrast-enhanced Ultrasonography in Diagnosis of Kidney Transplantation Rejection
Hua ZHANG ; Weixiang LIANG ; Jiangxiu YU ; Baomei YAN
Chinese Journal of Medical Imaging 2014;(9):678-680,685
Purpose To quantitatively analyze the blood perfusion parameters after kidney transplantation with acute and chronic rejection by using contrast-enhanced ultrasonography (CEUS), in order to provide rapid assessment for rejection prognosis. Materials and Methods Thirty-one patients with kidney transplantation were followed up and divided into three groups according to clinical symptoms and graft biopsy results:normal group (n=6), acute rejection group (n=12) and chronic rejection group (n=13). CEUS was carried out to assess renal microcirculation perfusion. Results CEUS showed that the normal renal parenchyma was enhanced homogeneously whilst the rejected parenchyma was enhanced heterogeneously. The differences of area under the curve among the three groups was statistically signiifcant (F=37.102, P<0.01), with normal group >chronic rejection group > acute rejection group. The peak intensity in normal group was higher than that in acute rejection group (P<0.01), but it showed no difference with that in chronic group. In acute and chronic rejection groups, the arrival time and the time to peak intensity in interlobar artery and cortex were all later than those in normal group (P<0.05). The ascending slope of TIC in segmental artery, interlobar artery and cortex in the two rejection groups was both signiifcantly less than that in normal group. Conclusion CEUS is a valuable diagnostic tool in the evaluation of microcirculation perfusion in kidney graft, and can provide important reference for the prognosis of acute and chronic rejection after kidney transplantation.
8.Current condition and research progress of self-management in patients after percutaneous coronary intervention
Yu YAN ; Yi YANG ; Qiuwen NONG ; Ting HUANG ; Zhene LIANG
Chinese Journal of cardiovascular Rehabilitation Medicine 2017;26(3):351-353
After percutaneous coronary intervention (PCI) was performed,since CHD risk factors still exist, coronary restenosis rate remains high.Therefore, self-management after PCI is very important.The present article made a review on current condition and research progress of self-management in patients after PCI, aiming at providing reliable evidence for rehabilitation after PCI.
9.Mixed thyroid malignant tumors
Liang HUANG ; Yang YU ; Yan ZHANG ; Ming GAO
Chinese Journal of General Surgery 2009;24(7):564-567
Objective To study the diagnosis, treatment and prognosis of mixed thyroid malignant tumors. Methods Clinical data of 7 cases with merged different histological types of thyroid malignant tumor treated from January 1977 to December 2006 were retrospectively analyzed. Results Merged different histologic types of thyroid malignant tumor accounted for 0. 14% of all thyroid malignant tumors treated during this period. Preoperative imaging and laboratory data had no specific value in the diagnosis of this merged different histologic types of thyroid malignant tumors. Radical resection in combination of hormonal therapy and 131I radiotherapy achieved a satisfactory result, though thyroid malignant tumor combined with thyroid cancer usually predict a poor prognosis. Conclusions Merged different histologic types of thyroid malignant tumor is a rare clinical entity, with the pathogenesis being obscure and no consensus of opinion on its nomenclature. The prognosis depends on the highest ~ade among an individual group of malignant tumors.
10.Effects of stellate ganglion block on erythrocyte immunity in patients with acute cerebral infarction
Chunjing HE ; Qian YU ; Yaping FENG ; Daiyi LIANG ; Yan RAN
Chinese Journal of Anesthesiology 2010;30(z1):46-49
Objective To investigate the effects of stellate ganglion block (SGB) on erythrocyte immunity in patients with acute cerebral infarction.Methods Twenty-four patients (13 male, 11 female) who developed acute cerebral infarction for less than 3 days were randomly divided into 2 groups (n=12each): Group A receiving traditional treatment and Group B receiving traditional treatment + SGB.The patients ranged in age from 51 to 64 yr and weighed 52-71 kg. All patients received intravenous 5% glucose 25 ml plus citicoline sodium 1.0 g and sodium ozagrel injectio 250 ml daily for 10 days in addition to dehydration and effective control of complications and intracranial pressure. Group B received SGB on one side alternatively with 1% licocaine 10 mi once a day for 10 days. Fasting venous blood samples were taken in the early mornings of the day before treatment (baseline, T1 ) and the 1st, 5th and 10th day of treatment (T2-4) for determination of the plasma MDA concentration and SOD activity, erythrocyte C3b receptor rosette rate (RBC-C3bRR) and RBC immune complex rosette rate (RBC-ICR) and Ne+-K+-ATPase activity in erythrocyte membrane.Results The plasma MDA concentration and RBC-ICR were significantly decreased during treatment es compared with the baselines at T1 in both groups (P<0.05 or 0.01), but were significantly lower in Group B than in Group A (P<0.05 or 0.01 ).The activities of plasma SOD and Na+ -K+ -ATPase in erythrocyte membrane and RBC-C3bRR were significantly increased during treatment as compared with the baselines at T1 and were significantly higher in Group B than in Group A.Conclusion SGB combined with traditional treatment can increase the activities of plasma SOD and Na+ -K+ -ATPase in erythrocyte membrane, inhibit production of oxygen free radicals and enhance RBC immune function in patients with acute cerebral infarction.