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.
3.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.
4.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.
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.
7.Application Analysis on Decoction Pieces in Beijing Traditional Chinese Medicine Hospital from 2010 to 2013
Yan CHEN ; Bin LI ; Yu LIU ; Lin LIANG
Chinese Journal of Information on Traditional Chinese Medicine 2015;(2):113-115
Objective To investigate the use condition of Chinese decoction pieces in Beijing TCM Hospital of Capital Medical University. Methods Excel was used to analyze the data collected from the hospital, including transfer, storage, and income and expense data of decoction pieces from 2010 to 2013. Results Workload of transfer of Chinese decoction pieces increased every year, and the staff were heavily loaded. Compared with western medicine and Chinese patent medicine, Chinese decoction pieces require more manpower. There is no compensation policy for all aspects of loss in Chinese patent medicine. Hospitals are responsible for relevant disposable consumable items. Conclusion The management of Chinese decoction pieces in our hospital is in an awkward situation, which calls for countermeasures as soon as possible.
8.Preliminary study of high frequency ultrasound screening for hyperuricemia in patients with joint disease
Xiuzhen HE ; Qiaorong LIANG ; Shouyi YU ; Yan YANG ; Yuncong CHEN
Chinese Journal of Rheumatology 2013;17(8):557-559
Objective To explore the value of high frequency ultrasonography for the early diagnosis of hyperuricemia in patients with joint disease.Methods Ninety-eight patients with hyperuricemia and 100 healthy persons,according to with or without history of acute gout attack,were divided into symptomatic group,asymptomatic group and control group,whose first metatarsophalangeal joint,ankle and knee were examined by high frequency ultrasonography.Chi-square test was used for statistical analysis.Results ① The joint lesions detection rate in the symptomatic group,asymptomatic group and the control group was 57% (13/23),16% (12/75),0 (0/100) respectively.There was statistical significant difference between the symptomatic group and the asymptomatic group (x2=9.69,P<0.05).② The symptomatic group had 29 joint involvement (29/138),including 25 at the first metatarsophalangeal joint,3 at the ankle and one at the knee joint.The asymptomatic group had 14 joints involved (14/450),which were all located at the first metatarsophalangeal joint.③ The sonographic appearance of the two groups of arthropathy were synovial thickening,effusion,crystal deposition andbone erosion.The symptoms group presented as tophi,and increased synovial blood flow.Conclusion Patients with asymptomatic hyperuricemia may have joint diseases.The joint disease detection rate of the symptomatic group is higher than that of the asymptomatic group.High frequency ultrasonography can be used a conventional imaging method for the screening of patients with hyperuricemia joint disease.The focus of the screening of asymptomatic patients is the first metatarsophalangeal joint.The focus of the screening of symptomatic patients is the first metatarsophalangeal joint and the joints that were attacked in the past history.
9.Clinical Observation of Spectral Entropy Monitoring as A Measure of the Conscious State during Neurosurgical Operation
Yan ZHANG ; Yu LIANG ; Xiushan ZHANG ; Haigen LIU
Tianjin Medical Journal 2010;38(3):195-197
Objective:To evaluate the clinical effectiveness of conscious state-monitoring device based on time-frequency-balanced spectral entropy of electroencephalogram monitoring during neumsurgical operation.Methods:Sixty-six patients scheduled for elective neumsurgical operation were randomly divided into two groups:control group(Ⅰ group,n=30)and observation group(Ⅱ group,n=36).Patients in control group were titrated to maintain adequate conscious state based on the monitoring of stable intraoperative hemodynamics.In the observation group,anesthetic was given to adjust to maintain the spectral entropy value to keep the adequate conscious state during operation.The values of mean arterial blood pressure (MAP),heart rate(HR),state entropy(SE)and response entropy(RE)were recorded before anesthesia induction,skin incision,at 3 min after intubation,at brain tissue dissection and during the last 10 min.And the parameters before recovery from anesthesia,the total amount of narcotic given were also recorded.Results:The values of SE and RE were significantly lower in the observation group than those in control group at 3 rain after intubation,skin incision,at brain tissue dissection and during the last 10 min,whereas MAP and HR were higher in control group compared with those of the observation group(P < 0.05 or P <0.01).The recovery index including recovery time of spontaneous breathing,time to eye opening and extubation was significantly decreased in observation group than that in the control group(P < 0.05 or P < 0.01).The total amount of narcotic consumption was significantly higher in the control group than that of the observation group(P < 0.01).Conclusion:During neumsurgical operation,the monitoring of conscious state by spectral entropy is more effective and better than hemodynamic standard practice.
10.DETECTION OF ORIENTIA TSUTSUGAMUSHI DNA FROM THE SPECIMENS OF BLOOD CLOT AND SPLEEN COLLECTED BOTH FROM EXPERIMENTAL INFECTED MICE AND FIELD RODENTS BY NESTED POLYMERASE CHAIN REACTION
Yansheng YAN ; Liang CHENG ; Shi HE ; Enshu YU
Chinese Journal of Zoonoses 2000;(6):39-41
AimTo evaluate the efficiency of gene amplification technique used in detecting the specimens colleted from rodents to identify natural epidemic foci of scrub typlus. MethodMice of Kunining strain were experimentally infected by a certain amount of Oriential tsutsugamushi. The specimens of blood clot and spleen from the infected animals were detected by nested polymerase chain reaction(nPCR)specific to O. T sutsugamush at the day 3,6 and 9 of post-infection. Then the technique was used for detection of samples collected from field. As an infected index ,the specimen was considered to be positive only if a 88-bp DNA fragment from Sta 58kDa gene of O. Tsutsugamushi could be produced. According to the study ,it was estimated whether or not that the sampling area is a natural epidemic focus of the disease. ResultsThe specimens of both blood clot and spleen from the mica at day 3 of post-infecction showed negative to the specific PCR product ,but positive when detected at day 6 and hereafter. Of 111 spleen samples from the field collections in the northwest of Fujian province,one was positive, and another positive sample was in the 29 blood clots from Jiangxi province. It is demonstrated that these areas have been the natural epidemic foci. Conclusion The nPCR method is of highly sensitive and specific to be used in the etiologic study on specimens from field rats.