1.Development of Loop-mediated Isothermal Amplification (LAMP) Method for Detection of Vibrio parahaemolyticus
Qian XU ; Xiao-Hong SUN ; Yong ZHAO ; Ying-Jie PAN ;
China Biotechnology 2006;0(12):-
Vibrio parahaemolyticus has been considered as one of the most important foodborne bacterial pathogens.The loop-mediated isothermal amplification(LAMP) that amplifies DNA with high specificity and rapidity under an isothermal condition was applied for rapid detection of this pathogen for the first time.A set of four primers,two outer and two inner primers,was designed specifically to recognize the thermolabile hemolysin gene(tlh) of V.parahaemolyticus.The LAMP reaction mix was optimized.The most optimal reaction temperature and time of the LAMP assay for the tlh gene were 60℃ and 60min,respectively.Genomic DNAs from 28 bacterial strains including 14 V.parahaemolyticus strains were amplified using LAMP,and no amplicon was observed in other bacterial strains.The detection limit of this LAMP assay was around 90 fg of V.parahaemolyticus genomic DNA and 24 colonies forming units for pure cultures.In addition,this method was applied to detect artificially contaminated food samples,and the detection limit was 89 cfu/g for non-cultured artificially contaminated food samples.These results suggested that detection of V.parahaemolyticus by LAMP is an effective and low-cost procedure with high specificity and sensitivity that requires no specialized equipment.This assay is expected to become a valuable tool for rapid detection and identification of V.parahaemolyticus.
2.Placement of a Long Intestinal Tube in Patients with Early Postoperative Small Bowel Obstruction under Fluoroscopic Guidance.
Zhi-wei WANG ; Xiao-guang LI ; Jie PAN ; Ning YANG ; Hai-feng SHI ; Zheng-yu JIN
Chinese Medical Sciences Journal 2015;30(3):156-161
OBJECTIVETo investigate the placement of a long tube into the small intestine under fluoroscopic guidance and to evaluate its decompression effect on early postoperative small bowel obstruction (EPSBO).
METHODSFifty-four patients with EPSBO requiring decompression between April 2010 and July 2014 were enrolled in the study. Insertion of a long tube was guided by fluoroscopy. We first used the guide wire to pass the pylorus and then used the 10 Fr feeding tube as an exchangeable tube to put the superstiff wire into the duodenum. Finally the long tube could be passed over the guide wire through the pylorus into the intestine. The total procedure time, the radiation exposure time, and the incidence of complications were evaluated.
RESULTSThe long tubes passed into the jejunum on initial insertion for all patients, so the success rate of this technique was 100%. The long tube was inserted into ileum in 18 patients. The mean total procedure time was 34.4 ± 8.6 minutes, and the mean radiation exposure time 18.9 ± 6.8 minutes. A total of 47 patients (87%) experienced full recovery following long-tube decompression and without the need for surgical intervention.
CONCLUSIONSUsing the wire-exchange technique, it is easy to place a long tube into the small bowel under fluoroscopic guidance. This decompression method is safe and effective for management of EPSBO.
Adult ; Aged ; Decompression, Surgical ; methods ; Female ; Fluoroscopy ; Humans ; Intestinal Obstruction ; surgery ; Male ; Middle Aged ; Postoperative Complications ; surgery ; Retrospective Studies
3.Research on body composition of soldiers in the graduating class of armed services colleges.
Chun-Jie PAN ; Xiao-Dong LIU ; Hui ZHANG ; Ying-Chun BAO
Chinese Journal of Applied Physiology 2014;30(4):311-313
OBJECTIVETo master the distribution of male and female soldiers' body composition with bioelectrical impedance analysis (BIA) method in order to provide data for building up the standards in comparison with those of ordinary residents.
METHODSA cluster stratified sample of 5968 graduated soldiers among different armed services colleges was measured by BIA. Five percent-95% distributions with in the upper and lower limits were established on basis of the above result. The crosswise comparison was also performed.
RESULTS(1) Five percent-95% reference ranges of people in the graduating class of armed services colleges were body fat percentage (BF%): 10.30%-20.70% (male) and 19.20%-30.10% (female), body mass index (BMI): 19.30-25.70 (male) and 18.00-23.99 (female), lean body mass percentage (LBM%): 79.27%-86.69% (male) and 69.89%-80.69% (female), muscle percentage (M%): 74.24%-83.96% (male) and 65.23%-75.27% (female), bone percentage (B%): 5.01% 5.77% (male) and 4.65%-5.51% (female). (2) Soldiers in the graduating class of armed services colleges have less BF%, more B% and M% than those of ordinary residents in the same age and the same sex.
CONCLUSION(1) Soldiers in the graduating class of armed services colleges have better body composition than that of ordinary residents in the same age and the same sex; (2) Standard of body composition for soldiers in the graduating class of armed services colleges should be different from that of ordinary residents, a new standard should be built up.
Body Composition ; Female ; Humans ; Male ; Military Personnel ; Young Adult
4.Epithelial-to-mesenchymal transdifferentiation of peritoneal mesothelial cells mediated by oxidative stress in peritoneal fibrosis rats
Shaobin DUAN ; Jie YU ; Qing LIU ; Yuhui WANG ; Peng PAN ; Li XIAO ; Guanghui LING ; Fuyou LIU
Journal of Central South University(Medical Sciences) 2011;36(1):34-43
Objective To investigate the role of oxidative stress in the epithelial-to-mesenchymal transdifferentiation (EMT) of peritoneal mesothelial cells in rat model of peritoneal fibrosis and the effect of probucol on peritoneal fibrosis. Methods The rat model of peritoneal fibrosis was induced by 4.25% high glucose peritoneal dialysis fluid (PDF). The rats were randomly divided into 4 groups:the control group, the saline group, the peritoneal fibrosis group, and the probucol group. A 4 hour peritoneal equilibration test (PET) was performed 4 weeks later. The peritoneal function and net ultrafiltration (UF) volume were determined. The level of malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) in peritoneal tissue were examined. The histology of peritoneal membrane was evaluated by light microscopy. E-cadherin and α-smooth muscle actin (α-SMA) protein expression was evaluated by immunohistochemical method and Western blot.Results The mesothelial cells were detached from peritoneal membrane in peritoneal firbosis rats. Comparing with the control rats, the thickness of visceral peritoneum, the level of MDA, and the-SMA protein expression were increased while the net ultrafiltration volume, the level of GSH-Px and E-cadherin protein expression were decreased in peritoneal firbosis rats. All these changes were reversed in the rats treated with probucol.Conclusion Oxidative stress plays an important role in transdifferentiation of peritoneal mesothelial cell in the peritoneal fibrosis rats. Probucol can improve structure and function of peritoneum, and partially reverse the EMT by reducing the oxidative stress.
5.Cliaical study of altering immunity for Chuanxiongqin ulinastain and thymosin α1 on sepsis patients
Yong PAN ; Jie XV ; Lanzhi ZHAO ; Bin ZHANG ; Yinjie CHEN ; Qiusheng XIAO ; Xiu QIN
Chinese Journal of Primary Medicine and Pharmacy 2008;15(8):1263-1264
Objective To investigate the effect of microcireulation blood flow and altering immunity by Chuanxiongqin,ulinastain and thymosin α1 on sepsis patients.Methods 90 patients were randomly divided into 3 groups(n=30),namely ICU group,Chuanxiongqin group,ulinastain and thymosin α1 group.HLA-DR/CD14+and IL-6,TNF-α,Lac,DD were measured.Results (1)DD showed no significant difference at every time point between ICU group and ulinastain+thymosin α1 group(P>0.05).DD decreased in Chuanxiongqin group,and was significantly different from the others on the third day.(2)Lac unchanged significantly at every time point in ICU group(P>0.05).Lac in Chuanxiongqin group and ulinastain+thymeain α1 group tended to decrease,and was statistically different from ICU group on the second day.(3)IL-6 and TNF-α tended to increase at every time point in ICU group(P<0.05).In ulinastain+thymosin α1 group,IL-6 and TNF-α returned to the level before treatment,HLA-DR/(D14+increased significantly,and was higher than Chuanxiongqin group and ICU group statistically.Conclusion Chuanxiongqin could ameliorate circulation;ulinastsin and thymosin α1 could depress IL-6,TNF-α.So ulinastain and thymosin α1 might protect the immunity of sepsis patients.
6.Dynamic predictive modeling of extraction process for red ginseng using near-infrared spectroscopy.
Jie-Qiang ZHU ; Wan-Fang PAN ; Yi ZHONG ; Xiao-Hui FAN ; Li-Yuan KANG ; Zheng LI
China Journal of Chinese Materia Medica 2014;39(14):2660-2664
It is the objective of this study to develop dynamic predictive model for the extraction process of red Ginseng using NIR spectroscopy. NIR spectroscopy was collected online and PLSR models were developed for total quantity of ginsenosides. The performance of NIR prediction model achieved R, RMSEC, RMSEP of 0.996 09, 0.018 9, 0.016 8, respectively. A first order dynamic mass transfer model was combined with NIR prediction of the quality indicator to predict the trajectory of the extraction process based upon the initial 3 or 4 data points. The results showed good agreement with actual measurements indicating reasonable accuracy of the predictive model. It could potentially be used for advanced predictive control of the extraction process.
Chemical Fractionation
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methods
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Ginsenosides
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chemistry
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isolation & purification
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Models, Theoretical
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Panax
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chemistry
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Spectroscopy, Near-Infrared
7.Research Progress in the Mechanism of Neuropathic Pain and the Intervention Effect of Electroacupuncture
Ningfang PAN ; Junying DU ; Jie YU ; Ting XIAO ; Xiaoqin LE ; Junfan FANG ; Jianqiao FANG
Shanghai Journal of Acupuncture and Moxibustion 2017;36(1):103-107
Neuropathic pain is a chronic pain caused by primary nervous system damage and nerve dysfunction. Its pathogenesis is complex and diverse. It is difficult to treat clinically. In recent years, researchers used electroacupuncture to treat neuropathic pain and obtained a desirable effect. This article summarizes recent years’ studies on the main mechanisms of neuropathic pain and the intervention effect of electroacupuncture to provide reference for following studies on electroacupuncture treatment of neuropathic pain.
8.The effect of single chain variable fragment antibody to EC3-4 fragment of desmoglein 3 in a mouse model of pemphigus vulgaris
Xiao-Qing ZHAO ; Feng XUE ; Meng PAN ; Wei-Ping LI ; Jie ZHENG ;
Chinese Journal of Dermatology 1994;0(06):-
Objective To study the effect of single chain variable fragment (ScFv) antibody to EC3-4 fragment of desmoglein (Dsg) 3 in a mouse model of pemphigus vulgaris.Methods The ScFv an- tibody to EC3-4 fragment of Dsg-3 was injected subcutaneously into neonatal BALB/c mice at different time points;the mice were then evaluated clinically,histopathologically and by direct immunoflorescence exami- nation for the development of lesions.Results When injected alone,the ScFv antibody did not induce the appearance of key clinical features of pemphigus vulgaris.The antibody also did not prevent the develop- ment of pemphigus vulgaris features induced by sera of patients with pemphigus vulgaris,regardless of the time point of injection of ScFv antibody.Conclusion The ScFv antibody to EC3-4 fragment of Dsg-3 lacks pathogenicity in neonatal BALB/c mice,and also could not inhibit the development of lesions induced by sera from patients with pemphigus vulgaris.
9.Effect of ostrich acellular heterogeneous corneal stroma ectopic transplantation on T lymphocyte subsets in peripheral blood of BALB/c mice
Xianning, LIU ; Jie, WU ; Xianghua, XIAO ; Shiyin, PAN ; Yong, YIN ; Xiuping, ZHU
Chinese Journal of Experimental Ophthalmology 2014;32(7):617-620
Background Ostrich acellular corneal stroma possesses a similar constitution to human corneal stroma,so it is expected to become one of ideal biological corneal carriers.Objective This study was to investigate the immunogenicity of acellular stroma carrier of ostrich cornea and offer the information for the development of industrialization and clinical use of acellular stroma carrier of ostrich cornea.Methods Twenty fresh ostrich eyeballs and 20 porcine eyeballs were collected.Acellular corneal stroma carriers of ostriches and pigs were prepared using low temperature freezing joint enzyme digestion method and desiccant dehydration method and sterilized by cobalt-60 irradiation.The corneal stroma carriers were preserved using drying and dehydration method.Forty-five male BALB/c mice were randomly divided into the sham operation group,ostrich acellular corneal stroma group and porcine acellular corneal stroma group.Acellular corneal stroma carriers of ostriches and pigs(wet weight after rewatering was 10 mg/piece) were subcutaneously implanted to the back of BALB/c mice,respectively.Wound healing and inflammatory response on the operative site were observed,and phenotype and activating rate of CD4+,CD8+ and CD25+in peripheral blood of mice were dynamically detected 7,14 and 28 days after ectopically implantation of heterogeneous corneal stroma by immunofluorescence labeling and flow cytometry analysis.Results No swelling and exudation were seen in the skin of operative site of the mice with a good healing of wound after surgery.There were no significant differences in the activating rates of CD4+,CD8+ and CD25+ cells in the peripheral blood of mice among the sham operation group,the porcine acellular corneal stroma group and ostrich acellular corneal stroma group in the three time points after surgery(CD4+:F=0.74,P=0.50;F=0.39,P=0.05;F=3.46,P=0.58.CD8+:F=1.75,P=0.21 ;F=1.14,P=0.35;F=0.78,P=0.48.CD25+:F=0.52,P=0.61 ;F=3.53,P=0.62;F=2.42,P=0.13).Conclusions The ostrich acellular heterogeneous corneal stroma carrier possesses low immunogenicity.It is inferred that ostrich acellular corneal stroma carrier can be used in heterogeneous corneal transplantation.
10.Management of fungal corneal ulcer by corneal collagen cross-linking technique
Zhaoqin, HAO ; Jinxin, SONG ; Jie, WU ; Shiyin, PAN ; Xianning, LIU ; Yan, CHENG ; Xianghua, XIAO
Chinese Journal of Experimental Ophthalmology 2014;32(9):802-806
Background Fungal corneal ulcer is a visual-threatening eye disease,and drug therapy has a limiting efficacy.Corneal transplantation or eye enucleation sometimes is necessary to the severe patients.Corneal collagen cross-linking (CXL) is an effective method for some corneal diseases,but the study on CXL for fungal corneal ulcer is lack.Objective This study was to evaluate the clinical effectiveness and safety CXL for fungal corneal ulcer.Methods Fifteen 8-week-old healthy New Zealand white rabbits were used in this study and other 5 rabbits served as normal controls.Fungal corneal ulcer models were established in the right eyes of other 10 rabbits by infecting sickle bacteria liquid after corneal scratching and removing corneal epithelium,then decellularized ostrich corneal patch covered the defected cornea.The models were randomly divided into the non-treatment group and the CXL treatment group.Corneal lesions were examined under the slit lamp microscope every day,and cornea was pictured by laser scanning confocal microscope on the 3rd,7th,14th,21st and 28th day individually after CXL.All rabbits were sacrificed and corneal tissues were obtained 4 weeks after treatment,and the collagen fiber diameter and fibrocytes were observed under the scanning electron microscope.Results Fungal corneal ulcer models were successfully established by corneal scratching and decellularized ostrich cornea covering.The gray ulcer lesions and hypbae like bean pod were seen by slit lamp microscope and laser scanning confocal microscope 3 days after modeling.Corneal ulcer deepened and expanded 1 week later,and there were a large number of spore and hyphae criss-crossing as short rod in shallow stroma.Inflammatory cells were observed in corneal endothelial cells and ocular anterior chamber.In the CXL treatment group,the range of corneal epithelial deficiency was less than that in the nontreatment group on the 3rd,7th,14th,and 21st (all at P< 0.05).The diameters of collagen fibers were (24.6± 1.8) nm,(24.9 ± 1.9) nm and (43.0 ± 7.4) nm in the normal control group,non-treatment group and CXL treatment group,showing a significant difference among the 3 groups (F =27.05,P =0.00),and the collagen diameters were thicker in the CXL treatment group than those in the normal control group and non-treatment group (t =5.40,-5.30,both at P<0.05),and fibrocytes were seen among the collagen fibers.No significant difference was found in the collagen diameters between the non-treatment group and normal control group,and the fibrocytes were less in the non-treatment group.Conclusions CXL therapy can treat fungal corneal ulcer by enhancing collagen,promoting fibrocytes proliferation,suppressing fungus and inflammatory response and accelerating tissue repair.