1.Detection of Human Hepatocarcinoma Cell Line PLC/PRF/5 Genome Hepatitis b Virus DNA Integration with Alu-PCR
Journal of Medical Research 2015;44(4):88-90,41
Objective In this research with the method of Alu -PCR we investigate the integration of hepatitis B virus ( HBV) DNA in human hepatocarcinoma cell line (PLC/PRF/5) genome DNA.Methods We at first extracted the genome DNA from PLC/PRF/5 cells, and then the potential integration fragments of HBV DNA and human genome DNA were amplified with according to the Alu -PCR after three rounds PCR .The Alu-PCR amplification products were observated with agarose gel electrophoresis , then integration positive electrophoresis bandings were chipped and purified for nucleic acid sequencing .At last he bioinformatics information was acquired by blast online.Results Through agarose gel electrophoresis after Alu -PCR amplification, we got four potential integration bindings , among which we got three integration sequences of HBV DNA in human genome DNA .These integration sequences could be individually located in the human chromosome of 03p21.31, 05p15.33, 12q13 and 12-q14.1.Conclusion With Alu-PCR we can accurately measure the integration of HBV DNA in human genome DNA , and Alu-PCR can be a a convenience and economic method in the study of HBV DNA ′s integration in human genome DNA .
2.Effect of Opiod Pretreatments on Etomidate Induced Myoclonus:A Meta-analysis
Peng QIU ; Shuang QIU ; Youjing DONG
Journal of China Medical University 2016;45(4):318-323
Objective To assess the effect of opiod pretreatment on etomidate induced myoclonus. Methods The pertinent literatures were searched by two independent investigators from the following electronic databases:PubMed,Cochrane Library,EMbase,VIP,WanFang Data,and CHKD. Then the meta?analysis was performed by using RevMan 5.2 and STATA 12.0 software. Results A total of 24 RCTs involving 2 396 pa?tients were included for the study. Pretreatment ofμorκ?receptor agonists reduced myoclonus with RR=0.19(95%CI 0.14 to 0.27)and RR=0.22 (95%CI 0.12 to 0.40),respectively. Conclusion Pretreatment of opiods can reduce the incidence of etomidate induced myoclonus.
3.Effects of Dexmedetomidine on the Heart Rate Variability in Patients Undergoing Lower Limbs Operations with Application of Tourniquet
Yue SUN ; Shuang QIU ; Youjing DONG
Journal of China Medical University 2017;46(2):107-111,115
Objective To evaluate the effects of dexmedetomidine on the hemodynamics and the heart rate variability(HRV)in patients under-going lower limbs operations with application of tourniquet. Methods Forty patients undergoing lower limbs operations with application of tourni-quet were randomized assigned to dexmedetomidine group(group D,n=20)or control group(group C,n=20). After combined spinal-epidural anesthesia,group D received a continuous infusion of dexmedetomidine(0.5μg/kg for 10 min for loading dose and followed by 0.2μg·kg-1·h-1) until tourniquet deflation. The control group received normal saline instead. Mean arterial pressure(MAP),heart rate(HR),saturation of pulse ox-imetry(SpO2),low frequency power(LF),high frequency power(HF)and LF to HF ratio(LF/HF)were recorded at regular time points:imme-diately before loading dose(T0),before tourniquet inflation(T1),15 min after tourniquet inflation(T2),30 min after tourniquet inflation(T3),45 min after tourniquet inflation(T4),60 min after tourniquet inflation(T5),1 min after tourniquet deflation(T6),5 min after tourniquet deflation (T7)and 10 min after tourniquet deflation(T8). Results Compared with T0,the MAP of group D significantly decreased at T6-T8(P<0.05). Compared with T0 and T1,the MAP of group C increased at T2-T5(P>0.05). Compared with T2-T5,the MAP of group C significantly decreased at T6(P<0.05). Compared with group C,the MAP of group D significantly decreased at T6 and T7(P<0.05). Compared with T0,the HR of group D significantly decreased at T1-T8(P<0.05). Compared with T0,the HR of group C had no significant change at T1-T5(P<0.05). Compared with T1-T5,the HR of group D and group C significantly increased at T6(P<0.05). Compared with group C,the HR of group D significantly decreased at T1-T4 and T6(P<0.05). Compared with T0,the SpO2 of group D and group C significantly decreased at T6(P<0.05). Compared with group C,the SpO2 of group D significantly decreased at T1-T3(P<0.05). Compared with T6,LF of group D and group C significantly increased at T7(P<0.05). LF were comparable between groups D and C(P>0.05). Compared with T0,the HF of group D significantly increased and the LF/HF of group D significantly decreased at T1-T4(P<0.05). Compared with group C,the HF of group D significantly increased and the LF/HF of group D significantly decreased at T1-T4(P<0.05). Conclusion The appropriate dose of dexmedetomidine(loading dose 0.5μg/kg and maintenance dose 0.2μg · kg-1 · h-1)can significantly increase vagal tone and improve cardiac sympathetic and parasympathetic balance during tourniquet appli-cation.
4.The profile of transient ischemic attacks in younger patients
Shuang QIU ; Song LUAN ; Yue DAI
Chinese Journal of Practical Internal Medicine 2001;0(06):-
Objective To evaluate the profile of transient ischemic attacks in younger patients compared with older patients effectively.Methods We study 75 younger patients(≤45 years)compared with 90 older patients(≥65 years).History of presenting transient ischemic attacks,etiology,clinical feature and lab investigation were compared on the basis of the above age groups.Results Overweight、hypercholesterolemia、insomnia and vascular disease family history being more common in the younger group and Hypertension,ischemic heart disease,long-term history of smoking being more common in the elder group.Diabetes and high serum uric acid occurred high frequently in both groups.Conclusion A significant correlation existed between etiologies contribution and adverse life custom,the early management should be take up to prevent TIA occurrence.
6. HPLC fingerprint and catalpol content determination of Rehmannia glutinosa
Chinese Traditional and Herbal Drugs 2019;50(20):5060-5063
Objective: The fingerprint of Rehmannia glutinosa was established by HPLC to provide scientific basis for improving the quality of R. glutinosa. Methods: The characteristic silicagel C18 column-Atlantis T3 was used, and the mobile phase was 0.1% phosphate water and acetonitrile by gradient elution. Absorption wavelength was 203 nm, and flow rate was 1.0 mL/mL with column temperature of 35 ℃. Setting catalpol as the control peak, 20 batches of R. glutinosa fingerprint was established. Fingerprint similarity evaluation software was used for data evaluation and determinating the catalpol content of 20 batches of R. glutinosa. Results: The standard contrast chromatographic fingerprint similarity of 20 batches R. glutinosa reached above 0.917. The 20 batches catalpol content was above 0.2%. Conclusion: The established fingerprint chromatography was proved that it had good precision, stability, and repeatability. The catalpol content determination met requirement which can avoid interference of saccharide compared with China Pharmacopoeia method and provide scientific reference for improving the quality of R. glutinosa.
7.Maturity and scientific research strength of medical subjects between PRC and India and between PRC and Japan according to the scale and production distribution of authors
Sisi CHEN ; Shuang XU ; Dan XU ; Yuhong QIU ; Jijun GUO
Chinese Journal of Medical Library and Information Science 2017;26(7):21-26
The maturity and scientific research strength of medical subjects between PRC and India and between PRC and Japan were demonstrated by comparing the scale and production distribution of international papers authors between PRC and India and between PRC and Japan according to the SCADC(2011), SCIE, InCites Statistical Analysis Database, non-regression fitting curve, K-S Test and Rnew fitting goodness, which showed that SCADC(2011) and SCIE displayed 53 specific medical subjects.Lotka distribution and author scale analysis showed that the maturity and scientific research strength of tropical medicine and pediatrics were lower while those of drug abuse, combined traditional and western medicine, primary health care, medical informatics, parasitology, audiology, speech and language pathology were higher in PRC than in Japan.
8.Determination of Related Substances of Bifonazole Raw Material by HPLC
Chunhui HAN ; Yilan QIU ; Shuang LUAN ; Xin GENG
China Pharmacy 2016;27(12):1713-1715
OBJECTIVE:To establish a method for the determination of related substances in bifonazole raw material. METH-ODS:HPLC method was performed on the column of Kromasil C18 with mobile phase of methanol-0.02 mol/L phosphoric acid(ad-justed pH to 7.5 with triethylamine)(70:30,V/V)at a flow rate of 1.0 ml/min,detection wavelength was 258 nm,volume injection was 20 μl,and the column temperature was 40 ℃. RESULTS:The linear range was 0.05-0.25 μg/ml for bifonazole(r=0.9996), 0.05-0.25 μg/ml for impurity A(bifonol)(r=0.9997)and 0.05-0.25 μg/ml for impurity B(4-C isomer)(r=0.9995);the detec-tion limits were 8.2 ng/ml,7.5 ng/ml and 8.4 ng/ml,and the quantification limits were 27.1 ng/ml,24.7 ng/ml and 27.8 ng/ml,re-spectively;RSDs of precision and reproducibility tests were lower than 1%;recovery of impurity B was 95.13%-101.29%(RSD=1.89%,n=9);both impurity A and impurity B were were detected in the 3 batches of samples. CONCLUSIONS:The method is accurate,sensitive and reproducible,and can be used for the determination of the related substances in bifonazole raw material.
9.SURFACE EXPRESSION AND COLOCALIZATION OF NMDA RECEPTOR AND AMPA RECEPTOR ON DENDRITE OF HIPPOCAMPAL NEURONS IN CULTURE
Shuang QIU ; Minzhen WANG ; Zhanyan FU ; Man HUANG ; Jianhon LUO
Acta Anatomica Sinica 1953;0(01):-
Objective To investigate the developmental profiles on surface expression and colocalization of NMDA receptor and AMPA receptor on dendritic structures in cultured hippocampal neurons of rats. Methods Two vectors expressing green fluorescent protein tagged GluR1 subunit(GFP-GluR1) and FLAG tagged NR2B subunit(FLAG-NR2B) were co-transfected into cultured hippocampal neurons at 5 days in vitro(DIV5).Surface expressed GFP-GluR1 containing AMPA receptor clusters and FLAG-NR2B containing NMDA receptor clusters were then labeled in living neurons by using anti-GFP primary antibody followed by Alex488-conjugated secondary antibody,and anti-FLAG primary antibody followed by Cy3-conjugated secondary antibody.Furthermore,distribution of the surface clusters was observed on the detailed dendritic structures visualized with co-expression of cyan fluorescent protein tagged actin(CFP-Actin). Results The numbers for GluR1 containing AMPA receptor clusters,NR2B containing NMDA receptor clusters,and colocalized receptor clusters per 100*!?m dendrite in DIV7 and DIV14 neurons were 59.2?5.6 and 74.8?3.1(P
10.A Rapid Method by Type-specific Multiplex PCR for Clostridium perfringens on Surface of Traumatic Tissues
Ying LIU ; Huiying ZHANG ; Yan WANG ; Xiaoxia AN ; Shuang QIU
Chinese Journal of Nosocomiology 2009;0(22):-
OBJECTIVE To establish a rapid detecting method for Clostridium perfringens on traumatic tissues by type-specific multiplex PCR.METHODS Established a simple and rapid method(TLS method) for purifying the DNA of genomes and plasmids in standard strains of C.perfringens wild strains on surface of open traumatic tissues and detected DNAs by type-specific multiplex PCR.RESULTS All types of C.perfringens could be detected by type-specific multiplex PCR.The sensitivity by PCR for type A of C.perfringens arrived at 7.5?103/ml and a whole run could finish within 5 h;the results by PCR entirely corresponded with those by cultureing.CONCLUSIONS New methods for purifying DNA of genomes and plasmids of C.perfringens are simple and rapid;there are high specificity and sensitivity for detecting DNA by multiplex PCR within short time,which can be practiced in clinical laboratory.