2.Establishment and performance evaluation of the quantitative detection for procalcitonin based on fluorescence immunochromatography
Qi FANG ; Xirong HUANG ; Kai LI ; Shixing TANG ; Jihua WANG
Chinese Journal of Laboratory Medicine 2012;(12):1102-1107
Objective To develop a quick quantitative detecting method for point of care testing (POCT) of human serum procalcitonin (PCT) by fluorescence immunochromatographic technology.Methods Applying a double-antibody sandwich immunofluorescent assay (one antibody coated on the nitrocellulose membrane and the other antibody labeled with fluorescent micropaticles) to develop a PCT quantitative detecting kit by immunochromatography technology.The kit was used to test PCT in 472 serum samples from suspected bacterial infection patients of Guangzhou Red Cross Hospital,including 240 male and 232 female patients.The methodology and diagnostic performance were evaluated in the aspects of linearity,precision,accuracy,specificity,stability experiments and comparison with foreign PCT detecting kits.Results The report range of the PCT quantitative diagnostic kit was 0.1-125.0 μg/L The coefficient of variation (CV)values of repeat 20 tests for low,median,and high concentration control samples respectively were all less than 15% and bias can be acceptable (P > 0.05).Common interfering substances in human serum specimens such as bilirubin (2.0 g/L),triglyceride (30.0 g/L) and cholesterol (15.0 g/L) were found no significant affect on quantitative detection of PCT.The shelf time of the PCT diagnostic kit should be longer than 12 months as the relative deviation of detected concentrations of 0.5,1.0,22.0,65.0 μg/L PCTcontrol sample can be controlled less than 20% within 14 months.Considering VIDAS BRAHMS PCT to be the standard quantitative test for PCT,472 serum samples were detected by both our kit and the control VIDAS BRAHMS PCT kit simultaneously,which showed high correlation (YVIDAS =0.180 + 1.006Xwondfo,R2 =0.988,P < 0.01) and low deviation (Z =-1.6,P > 0.05) without statistic significance between two methods.And the results of these two diagnostic kits showed good consistency as the area under curve of the receiver operating characteristic (ROC) of Wondfo-PCT at the three cut-off values (0.5,2.0,10.0 μg/L)were 0.997,0.994,0.998 respectively,P < 0.01,using diagnostic result of the control product as standard.Kappa values were 0.899,0.905,0.973 respectively.Conclusions The method of quantitative detection of PCT by fluorescence immunochromatography for POCT was established in this study.All the observed indicators reached the clinical diagnostic requirements and can be applied for the quick detection of clinical human serum PCT.
3.Role of FTY720 therapy in motor nerve function recovery after acute spinal cord injury in rats
Jianmin WANG ; Kai TANG ; Benchen XU ; Chengke WANG ; Qazi BASIT ; Fang LI ; Xuezheng ZHAO
Chinese Journal of Trauma 2013;29(10):1000-1007
Objective To investigate the effect and mechanism of FTY720 in protecting the motor nerve function in rats with acute spinal cord injury (ASCI).Methods A model of ASCI in the rat was established using Allen' s method.The rats were divided into five groups according to random number table:Group A (normal control group,n =6),Group B (sham group,n =27),Group C (ASCI group,n =27),Group D (FTY720 systemic treatment group,n =27) and Group E (FTY720 topical treatment group,n =27).After treatment,each group was observed for the changes of lymphocytes in peripheral blood and topical spinal cord tissue,BBB score,pathological changes of spinal cord tissue and expression intensity of glial fibrillary acidic protein (GFAP).Results After ASCI in rats,the infiltration of local T lymphocyte subpopulations CD3,CD4 and CD8 was promoted,but the number of peripheral blood T lymphocyte subpopulations CD3,CD4 and CD8 was decreased.After FTY720 therapy,both local and peripheral blood lymphocytes were reduced in number and the decrease of peripheral blood lymphocytes in Group E was between Groups C and D.BBB score,pathologic observation and GFAP staining showed a neuroprotective effect of FTY720 and better motor nerve function recovery in Group E than in Groups C and D.Conclusions FTY720 therapy facilitates motor nerve function recovery.Local FTY720 application reduces peripheral blood lymphocytes and lowers the incidence of infection.
4.Effect of Jinwu Jiangu Recipe on Expressions of NF-kappaB and IL-17 in Collagen Induced Arthritis Model Rats.
Wu-kai MA ; Dao-min LU ; Xue-ming YAO ; Ying HUANG ; Fang TANG ; Jiang LIANG ; Yang AN ; Jing ZHOU
Chinese Journal of Integrated Traditional and Western Medicine 2016;36(5):624-628
OBJECTIVETo explore the effect of Jinwu Jiangu Recipe (JJR) on the expression of synovial cells' nuclear factor-kappaB (NF-kappaB) and serum interleukin 17 (IL-17) in collagen induced arthritis (CIA) rats.
METHODSTotally 60 Wistar rats were randomly divided into 6 groups, i.e., the blank control group, the model group, high, middle, and low dose JJR treatment groups, and the tripterygium control group, 10 in each group. Except rats in the blank control group, CIA model was established in rats of the rest 5 groups. Then they were treated from the 7th day of modeling. After 4 weeks of medication they were sacrificed, serum collected, and synovium of joints were isolated. The expression of serum IL-17 was detected in synovium of joints by enzyme linked immunosorbent assay (ELISA). And the expression of NF-kappaB/P65, Ikappabetaalpha and NF-KappaB/P50 were detected by Western blot.
RESULTSCompared with the blank control group, the serum IL-17 level increased in the model group (P <0. 01). Compared with the model group, the serum IL-17 level obviously decreased in high and middle dose JJR groups and the tripterygium control group (P < 0.01). Results of Western blot showed, when compared with the blank control group, protein activities of NF-kappaB/P65 and NF-kappaB/P50 were significantly enhanced in the model group (P < 0.01). Compared with the model group, protein activities of NF-kappaB/P65 and NF-kappaB/P50 significantly decreased in high and middle dose JJR groups and the tripterygium control group (P < 0.05, P < 0.01). All indices mentioned above were higher in the low dose JJR group than in the tripterygium control group (P < 0.05, P < 0.01).
CONCLUSIONJJR could lower the expression of serum IL-17 in CIA model rats, and inhibit protein activities of NF-kappaB/P65 and NF-kappaB/P50.
Animals ; Arthritis, Experimental ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; chemistry ; Interleukin-17 ; blood ; NF-kappa B ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Synovial Membrane ; drug effects ; metabolism ; Tripterygium ; chemistry
5.Clinical application of miniature tissue transplantation in hand surgery
Guangrong FANG ; Xiaohen DIN ; Haiping TANG ; Shujian HOU ; Guanghai YUAN ; Yaping LIU ; Zhigang QU ; Kai JIANG ; Guoliang CHENG
Chinese Journal of Microsurgery 2009;32(5):356-359,illust 1
Objective To explore the concept,denomination and technique of miniature tissue transplantation in hand clinic. Methods In 537 cases,local skin or composite tissue defect in 756 thumbs or fingers were repaired with 4 kinds of miniature island flap and 10 kinds of miniature vascularized composite tissue.Some of them were created in clinic practice.The precess,main points and indication of the operation were expounded and compared. Results In 639 fingers covered with miniature island flap,the transferred flaps were surviverd completely in 621 fingers(97.2%),survivered partly in 11 fingers(1.7%)and necrosised in 7 fingers(1%).In 117 fingers repaired with miniature vascularized composite tissue,all the transplanted tissues were survivered.In all the injuried fingers,beautiful outlook were recovered and no fat and clumsy were found,two-point difference was 6-8 mm,effctive sensibility and good function were restored,and all the patients were satisfied. Conclusion Miniature tissue transplantation were good methods to cover the local defect in fingers or thumbs,worth of generalization and aplication in hand clinic,and have an important significance for improve the trauma repair quality and treatment level in hand surgery.
6.Experience with intravascular lead extraction.
Jian MA ; Kai TANG ; Fang-zheng WANG ; Shu ZHANG ; Cong-xin HUANG
Chinese Medical Journal 2005;118(14):1217-1219
Adult
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Aged
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Aged, 80 and over
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Female
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Humans
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Lead
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isolation & purification
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Male
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Middle Aged
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Pacemaker, Artificial
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adverse effects
7.Suppurative arthritis caused by Gemella morbillorum in a patient with rheumatoid arthritis of the knee.
Fang-Gen TANG ; Fen-Lian YUAN ; Jia-Kai LIU ; Quan LONG
Journal of Southern Medical University 2012;32(3):437-438
Rheumatoid arthritis of the knee is a common disease, but suppurative arthritis caused by Gemella morbillorum in the same joint is rare. We report a case of suppurative arthritis caused by Gemella morbillorum in a patient with rheumatoid arthritis. Because the infection symptoms was not typical, the diagnosis was delayed, and the delayed diagnosis and therapy led to a poor outcome of the patient.
Arthritis, Infectious
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complications
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Arthritis, Rheumatoid
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complications
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Female
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Gemella
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isolation & purification
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Gram-Positive Bacterial Infections
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complications
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Humans
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Knee Joint
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Middle Aged
8.Experience and discussion on the national standard Standardized Manipulation of Acupuncture and Moxibustion. Part 8: Intradermal Needle.
Ling LUO ; Cheng-Kai YUAN ; Hai-Yan YIN ; Fang ZENG ; Yong TANG ; Shu-Guang YU
Chinese Acupuncture & Moxibustion 2012;32(2):155-158
Standardized Manipulation of Acupuncture and Moxibustion Part 8: Intradermal Needle was compiled with the following principles. The compiling standard, technical features and clinic manipulations of intradermal needle were taken as the basic principle for compiling. Literature research, expert survey and clinic practice verification were applied as the drafting methods. The key issues were focused on the relationship between standardization and individualization, normalization and effectiveness, qualification and quantification. And the postural selection, reinforcing and reducing manipulations, fixing materials and embedding duration involved in intradermal needling were emphasized particularly. At the same time, details and the future way of thinking of intradermal needle were expounded in this article as well.
Acupuncture Therapy
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instrumentation
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standards
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China
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Humans
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Moxibustion
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standards
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Needles
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standards
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Reference Standards
9.Expression of WW domain containing oxidoreductase gene in cholangiocarcinoma and its effect on the biological behavior of cancer cell line RBE.
Qiang HUANG ; Kai ZHU ; Chen-hai LIU ; Fang XIE ; Peng XU ; Zhi-gang TANG
Chinese Journal of Surgery 2011;49(4):324-329
OBJECTIVETo study the effects of anti-oncogene WWOX on cell growth of cholangiocarcinoma.
METHODSThe expression of WWOX protein was detected with immunohistochemical method-SP in 54 patients with cholangiocarcinoma from July 2005 to May 2010 and 12 samples of normal bile duct tissues. The recombinant WWOX eukaryotic expression plasmid was introduced into RBE cells by liposome-mediated transfection and positive cell clones were selected and amplified. The mRNA and protein expressions in RBE cells stably transfected with WWOX were investigated by quantitative RT-PCR and Western Blot before and after transfection. Cell proliferation was tested by MTT, cell apoptosis was assessed by FCM, the alteration of mitochondria membrane potential (ΔΨm) was detected by JC-1 staining method, cell invasion was determined by Transwell chamber assay. The expression change of bcl-2, bax, FasL, caspase-3 mRNA and protein was detected by quantitative RT-PCR and Western Blot.
RESULTSThe expression of WWOX protein was significantly lower in cholangiocarcinoma than that in normal bile duct tissues and loss of WWOX protein expression was found in 40.7% of cholangiocarcinoma specimens (P < 0.05). RBE cells with stable transfection of WWOX were established. Quantitative RT-PCR showed that the expression of WWOX mRNA was significantly enhanced and Western Blot demonstrated that WWOX protein expression was markedly increased. MTT showed that WWOX gene transfection significantly decreased the proliferation of RBE cells (P < 0.05). FCM analysis showed that the apoptosis rate after transfection was significantly promoted [(1.1 ± 0.6)% vs. (1.7 ± 0.5)% vs. (35.2 ± 4.4)%, P < 0.01], JC-1 staining method indicated that the experimental group was loss of ΔΨm [(12.6 ± 1.9)% vs. (13.6 ± 1.8)% vs. (48.7 ± 2.9)%, P < 0.01], transwell chamber assay showed that the number of transfected cells that passed the transwell membrane was significantly less than those of control groups (77 ± 6 vs. 72 ± 8 vs. 48 ± 6, P < 0.01). Quantitative RT-PCR and Western blotting showed that the expression of bcl-2 mRNA and protein was markedly decreased and the expression of bax, caspase-3 were significantly increased. There was no significant change in the expression of FasL.
CONCLUSIONWWOX exerts its antitumor effect against proliferation through inducing cell apoptosis in cholangiocarcinoma.
Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Cholangiocarcinoma ; genetics ; metabolism ; pathology ; Genetic Vectors ; Humans ; Oxidoreductases ; genetics ; metabolism ; Plasmids ; genetics ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism ; WW Domain-Containing Oxidoreductase
10.Radiofrequency catheter ablation at the left coronary cusp in treatment of repetitive monomorphic tachycardia of the left ventricular outflow tract.
Kai TANG ; Jian MA ; Shu ZHANG ; Jian-min CHU ; Kui-jun ZHANG ; Fang-zheng WANG ; Xin CHEN
Chinese Medical Journal 2004;117(2):168-171
BACKGROUNDThe aim of this study was to clarify the electrocardiographic characteristics of repetitive monomorphic ventricular tachycardia (RMVT) originating from the left ventricular outflow tract, and to describe the results of treatment with radiofrequency catheter ablation (RFCA).
METHODSRoutine 12-lead surface electrocardiography and electrophysiological studies were performed on 11 RMVT patients with no organic heart disease, who were subsequently treated with RFCA directed at targets identified by pace mapping.
RESULTSThe surface electrocardiogram QRS characteristics of RMVT included an atypical left bundle branch block and right axis deviation, with a low amplitude "rs" or "rS" pattern in lead I, an "rS" or "RS" pattern in V1, and a precordial R wave transition zone in V2 or V3. In 1 patient, a small S wave was observed in V5. Using pace mapping techniques, we selected the left coronary cusp as the ablation target. RMVT was eliminated in all 11 patients immediately after radiofrequency energy delivered. During a follow-up of 13 +/- 7 months, RMVT recurred in only 1 patient.
CONCLUSIONRMVT originating from the left ventricular outflow tract has specific electrocardiographic characteristics, and can be successfully and safely cured using RFCA directed at the left coronary cusp.
Adolescent ; Adult ; Catheter Ablation ; Electrocardiography ; Female ; Humans ; Male ; Middle Aged ; Tachycardia, Ventricular ; physiopathology ; surgery ; Treatment Outcome