1. Asymmetric PCR method in generation of HBV ssDNA for pyrosequencing
Academic Journal of Xi'an Jiaotong University 2009;21(1):54-56
Objective: To explore the optimal primer ratio and concentration of asymmetric polymerase chain reaction (A-PCR) in producing hepatitis B virus (HBV) single-stranded DNA (ssDNA) for pyrosequencing. Methods: A-PCR was carried out to generate HBV ssDNA with forward to reverse primers of different ratios (50:1, 100:1) and concentrations (13.0 pmol/25μL and 0.14 pmol/25μL, 19.5 pmol/25μL and 0.21 pmol/25μL), and the product yield and quality were compared respectively. Results: The forward to reverse primer ratio of 50:1 provided better yield and concentration of 19.5 pmol/25μL and 0.21 pmol//25μL generated a clearer band. Conclusion: A simple and feasible method to produce HBV ssDNA for pyrosequencing in batch is established.
2. Asymmetric PCR method in generation of HBV ssDNA for pyrosequencing
Academic Journal of Xi'an Jiaotong University 2009;21(1):54-56
Objective: To explore the optimal primer ratio and concentration of asymmetric polymerase chain reaction (A-PCR) in producing hepatitis B virus (HBV) single-stranded DNA (ssDNA) for pyrosequencing. Methods: A-PCR was carried out to generate HBV ssDNA with forward to reverse primers of different ratios (50:1, 100:1) and concentrations (13.0 pmol/25μL and 0.14 pmol/25μL, 19.5 pmol/25μL and 0.21 pmol/25μL), and the product yield and quality were compared respectively. Results: The forward to reverse primer ratio of 50:1 provided better yield and concentration of 19.5 pmol/25μL and 0.21 pmol//25μL generated a clearer band. Conclusion: A simple and feasible method to produce HBV ssDNA for pyrosequencing in batch is established.
3.Urine Metabolites Changes in Acute Myocardial Infarction Rats via Metabolomic Analysis
Nian-Nian CHEN ; Jiao-Fang YU ; Peng WU ; Li LUO ; Ya-Qin BAI ; Li-Kai WANG ; Xiao-Qian LI ; Zhan-Peng LI ; Cai-Rong GAO ; Xiang-Jie GUO
Journal of Forensic Medicine 2024;40(3):227-236
Objective To screen biomarkers for forensic identification of acute myocardial infarction (AMI) by non-targeted metabolomic studies on changes of urine metabolites in rats with AMI.Methods The rat models of the sham surgery group,AMI group and hyperlipidemia+acute myocardial infarction (HAMI) group were established.Ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) was used to analyze the changes of urine metabolic spectrometry in AMI rats.Principal compo-nent analysis,partial least squares-discriminant analysis,and orthogonal partial least squares-discriminant analysis were used to screen differential metabolites.The MetaboAnalyst database was used to analyze the metabolic pathway enrichment and access the predictive ability of differential metabolites.Results A total of 40 and 61 differential metabolites associated with AMI and HAMI were screened,respec-tively.Among them,22 metabolites were common in both rat models.These small metabolites were mainly concentrated in the niacin and nicotinamide metabolic pathways.Within the 95% confidence in-terval,the area under the curve (AUC) values of receiver operator characteristic curve for N8-acetyl-spermidine,3-methylhistamine,and thymine were greater than 0.95.Conclusion N8-acetylspermidine,3-methylhistamine,and thymine can be used as potential biomarkers for AMI diagnosis,and abnormal metabolism in niacin and nicotinamide may be the main causes of AMI.This study can provide reference for the mechanism and causes of AMI identification.
4.Inhibiting effects of Achyranthes bidentata polysaccharide and Lycium barbarum polysaccharide on nonenzyme glycation in D-galactose induced mouse aging model.
Hong-Bin DENG ; Da-Peng CUI ; Jian-Ming JIANG ; Yan-Chun FENG ; Nian-Sheng CAI ; Dian-Dong LI
Biomedical and Environmental Sciences 2003;16(3):267-275
OBJECTIVETo investigate the inhibiting effects and mechanism of achyranthes bidentata polysaccharide (ABP) and lycium barbarum polysaccharide (LBP) on nonenzyme glycation in D-galactose induced mouse aging model.
METHODSSerum AGE levels were determined by AGE-ELISA, MTT method was used to determine lymphocyte proliferation, IL-2 activity was determined by a bioassay method. Spontaneous motor activity was used to detect mouse's neuromuscular movement, latency of step-through method was used to examine learning and memory abilities of mouse, colormetric assay was used to determine hydroxyproline concentration in mouse skin, pyrogallol autoxidation method was used to determine superoxide dismutase (SOD) activity of erythrocytes.
RESULTSDecreased levels of serum AGE, hydroxyproline concentration in mouse skin and spontaneous motor activity in D-galactose mouse aging model were detected after treated with ABP or LBP, while lymphocyte proliferation and IL-2 activity, learning and memory abilities, SOD activity of erythrocytes, were enhanced.
CONCLUSIONSABP and LBP could inhibit nonenzyme glycation in D-galactose induced mouse aging model in vivo and ABP has a better inhibiting effect than LBP.
Achyranthes ; chemistry ; Aging ; physiology ; Animals ; Disease Models, Animal ; Erythrocytes ; Female ; Galactose ; chemistry ; Learning ; Lycium ; chemistry ; Memory ; Mice ; Mice, Inbred C57BL ; Motor Activity ; Polysaccharides ; pharmacology ; Superoxide Dismutase ; pharmacology
5.Structural and functional changes of immune system in aging mouse induced by D-galactose.
Hong-Bin DENG ; Chun-Lei CHENG ; Da-Peng CUI ; Dian-Dong LI ; Li CUI ; Nian-Sheng CAI
Biomedical and Environmental Sciences 2006;19(6):432-438
OBJECTIVETo investigate the role of D-galactose, especially in the structural and functional changes of the immune system in aging.
METHODSSerum levels of advanced glycation end-products (AGE) were determined by ELISA method. Ultra-structures of thymus and spleen were detected by transmission electron microscopy. MTT method was used to determine the lymphocyte proliferation. IL-2 activity was determined by bioassay. Northern blot was used to detect the IL-2 mRNA levels.
RESULTSSerum AGE levels of D-galactose- (P < 0.01) and AGE-treated (P < 0.05) mice (n = 8) were increased significantly. The ultra-structures of thymus and spleen in D-galactose- and AGE-treated mice showed regressive changes similar to those in the aged control group. The lymphocyte mitogenesis and IL-2 activity of spleen were also decreased significantly (P < 0.01, n = 8). The change of IL-2 activity shown by Northern blot resulted from the change of mRNA expression. The AGE plus aminoguanidine group, however, showed no significant change in these parameters in comparison with the young control group (P < 0.01 or P < 0.05, n = 8).
CONCLUSIOND-galactose and AGE lead to a mimic regression change of aging in the immune system in vivo.
Aging ; drug effects ; immunology ; Animals ; Cell Proliferation ; drug effects ; Galactose ; pharmacology ; Glycation End Products, Advanced ; blood ; Interleukin-2 ; metabolism ; Lymphocytes ; drug effects ; immunology ; Mice ; Microscopy, Electron, Transmission ; RNA, Messenger ; metabolism ; Spleen ; drug effects ; immunology ; ultrastructure ; Thymus Gland ; drug effects ; immunology ; ultrastructure
6.Transtrochateric rotational osteotomy for osteonecrosis of the femoral head.
Nian-fei ZHANG ; Zi-rong LI ; Lian-fa YANG ; Peng LIN ; Zhen-cai SHI ; Zhong-shi LI ; Wei SUN
Chinese Journal of Surgery 2004;42(24):1477-1480
OBJECTIVETo explore the indications, operative technique and clinical results of the transtrochanteric rotational osteotomy (TRO) for osteonecrosis of the femoral head (ONFH).
METHODSNineteen patients (23 hips) with ONFH underwent TRO from October 1992 to May 2001 were reviewed. There were 14 males and 5 females. The age was ranging from 22 to 43 years old with an average age of 33.4 years old. The etiology were as followed: alcoholism 10 cases (12 hips); steroid 6 cases (7 hips); trauma 2 cases (2 hips); caisson disease 1 case (2 hips). Ficat stage: II 8 cases 8 hips; III 11 cases 15 hips. Harris hip score 46 to 74 with mean score 56 preoperatively. In order to determine the location and extent of osteonecrosis, A-P view and accurate lateral view (the film should be placed parallel to the femoral neck with the hip in precisely 90 degrees and flexion 45 degrees of abduction and neutral rotation) were taken in both hips preoperatively. The surgical technique described by Sugioka were used in all patients.
RESULTSSeventeen patients (19 hips) were followed up ranging from 18 to 11 years with mean 54 months. The Harris hip score was from 55 to 94 with mean 80.5. Fourteen hips score more than 80 and good to excellent rate was 73.2%. Harris score were less than 79 in 5 cases and 4 hips of these 5 hips were revised by total hip arthroplasty. The rotation angle was from 55 degrees to 80 degrees with mean 60 degrees .
COMPLICATIONSThe circumflex femoral medial artery injury was found in 1 hip; subtrochanteric fracture was found in 2 hips and 1 osteotomy site nonunion occurred.
CONCLUSIONTRO can be used to treat ONFH. The indications include: (1) younger patients (less than 45 years old). (2) Ficat stage II and III with more than one third intact articular surface of the femoral head. The better results could be obtained using posterior rotation and lager rotational angle. The Sugioka osteotomy should be used carefully because of its complicated surgical technique.
Adult ; Female ; Femur ; surgery ; Femur Head Necrosis ; surgery ; Follow-Up Studies ; Humans ; Male ; Osteotomy ; methods ; Treatment Outcome
7.Visualization Analysis of Artificial Intelligence Literature in Forensic Research
Yi-Ming DONG ; Chun-Mei ZHAO ; Nian-Nian CHEN ; Li LUO ; Zhan-Peng LI ; Li-Kai WANG ; Xiao-Qian LI ; Ting-Gan REN ; Cai-Rong GAO ; Xiang-Jie GUO
Journal of Forensic Medicine 2024;40(1):1-14
Objective To analyze the literature on artificial intelligence in forensic research from 2012 to 2022 in the Web of Science Core Collection Database,to explore research hotspots and developmen-tal trends.Methods A total of 736 articles on artificial intelligence in forensic medicine in the Web of Science Core Collection Database from 2012 to 2022 were visualized and analyzed through the litera-ture measuring tool CiteSpace.The authors,institution,country(region),title,journal,keywords,cited references and other information of relevant literatures were analyzed.Results A total of 736 articles published in 220 journals by 355 authors from 289 institutions in 69 countries(regions)were identi-fied,with the number of articles published showing an increasing trend year by year.Among them,the United States had the highest number of publications and China ranked the second.Academy of Forensic Science had the highest number of publications among the institutions.Forensic Science Inter-national,Journal of Forensic Sciences,International Journal of Legal Medicine ranked high in publica-tion and citation frequency.Through the analysis of keywords,it was found that the research hotspots of artificial intelligence in the forensic field mainly focused on the use of artificial intelligence technol-ogy for sex and age estimation,cause of death analysis,postmortem interval estimation,individual identification and so on.Conclusion It is necessary to pay attention to international and institutional cooperation and to strengthen the cross-disciplinary research.Exploring the combination of advanced ar-tificial intelligence technologies with forensic research will be a hotspot and direction for future re-search.
8.Progress in antiviral strategies targeting influenza virus hemagglutinin
Ming CAI ; Xuedan LI ; Liang SHAO ; Feixia PENG ; Zhegang ZHANG ; Bo LIU ; Xuanxuan NIAN ; Jiayou ZHANG ; Hui CAO ; Xiaoming YANG
Chinese Journal of Microbiology and Immunology 2023;43(11):850-859
Influenza is a worldwide infectious disease caused by influenza virus. It has posed great challenges on public health and social stability since 1918. At present, vaccination is the most effective way to prevent and control influenza epidemics. Broad-spectrum antiviral drugs and neutralizing antibodies against influenza virus have been widely studied in recent years. Hemagglutinin (HA), which is on the surface of influenza virus, plays an important role in the stage of viral invasion into host cells. It is the main effective antigenic component of current influenza vaccines, as well as the main target of broad-spectrum neutralizing antibodies and broad-spectrum antiviral drugs. This review summarized the progress in the development of novel influenza vaccines, neutralizing antibodies, and antiviral drugs based on influenza virus HA, as well as other prevention and control measures, hoping to present new ideas for future influenza prevention and control.
9.Screening Biomarkers of Sudden Coronary Death Based on mRNA Expression Profile of Rat Myocardial Tissues.
Xiang-Jie GUO ; Hao LI ; Ya-Qin BAI ; Peng WU ; Chun-Mei ZHAO ; Yi-Ming DONG ; Nian-Nian CHEN ; Ke-Ming YUN ; Cai-Rong GAO
Journal of Forensic Medicine 2022;38(4):443-451
OBJECTIVES:
To explore the differential expression of messenger RNA (mRNA) in myocardial tissues of rats with sudden coronary death (SCD), and to provide ideas for the forensic identification of SCD.
METHODS:
The rat SCD model was established, and the transcriptome sequencing was performed by next-generation sequencing technology. Differentially expressed genes (DEGs) in myocardial tissues of SCD rats were screened by using the R package limma. A protein-protein interaction (PPI) network was constructed by using the STRING database and Cytoscape 3.8.2 on DEG, and hub genes were screened based on cytoHubba plug-in. Finally, the R package clusterProfiler was used to analyze the biological function and signal pathway enrichment of the selected DEG.
RESULTS:
A total of 177 DEGs were associated with SCD and were mainly involved in the renin-angiotensin system and PI3K-Akt signaling pathway. The genes including angiotensinogen (AGT), complement component 4a (C4a), Fos proto-oncogene (FOS) and others played key roles in the development of SCD.
CONCLUSIONS
Genes such as AGT, C4a, FOS and other genes are expected to be potential biomarkers for forensic identification of SCD. The study based on mRNA expression profile can provide a reference for forensic identification of SCD.
Rats
;
Animals
;
RNA, Messenger/genetics*
;
Gene Regulatory Networks
;
Gene Expression Profiling
;
Phosphatidylinositol 3-Kinases/genetics*
;
Biomarkers
10.Herbal acupoint sticking combined with electroacupuncture therapy in the treatment of Bell's palsy: a randomized controlled trial.
Qi-Hua QI ; Shan-Shan NI ; You-Lan WANG ; Kai PENG ; He-Nian QU ; Cai-Hua YANG ; Jin WANG ; Wei XI
Chinese Acupuncture & Moxibustion 2013;33(11):965-969
OBJECTIVETo observe the clinical efficacy and safety of electroacupuncture (EA) combined with herbal acupoint sticking in the treatment of Bell's palsy and provide optimizations for the clinic.
METHODSOne hundred and two cases of Bell's palsy were randomized into an EA combined with herbal acupoint sticking group (group A, 50 cases) and an EA group (group B, 52 cases), EA at Cuanzhu (BL 2), Yangbai (GB 14), Taiyang (EX-HN 5), Quanliao (SI 18),Xiaguan (ST 7), Yingxiang (LI 20), etc. were applied in both groups and "facial paralys No.I " was applied at Yifeng (TE 17) in group A, once daily and 10 times totally were needed. The score of facial nerve function, clinical efficacy were compared before and after treatment. At 1 and 3 month follow up visit, the quality of life scale( WHOQOL-BREF) and the occurrence of complication were observed.
RESULTSThe scores of facial nerve function in group A and group B were all significantly improved compared with those before treatment (48. 2+/- 2. 9 vs 25. 7 +/- 4. 9, 45. 9 +/- 6. 2 vs 25. 8 +/- 5. 5, both P
CONCLUSIONCompared with EA, the combination of EA and acupoint sticking therapy for Bell's palsy cannot only improve the clinical efficacy and reduce the occurrence of complication but also reliable without any side effect.
Acupuncture Points ; Administration, Cutaneous ; Adolescent ; Adult ; Aged ; Bell Palsy ; drug therapy ; therapy ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult