1.Prokaryotic expression of human cTnI and preparation of anti-cTnI monoclonal antibody
Chinese Journal of Laboratory Medicine 2003;0(11):-
Objective To prepare monoclonal antibody (mAb) against recombinant human cardiac troponin I(cTnI).Methods The full-length gene encoding human cardiac troponin I (cTnI) was synthesized chemically and inserted into expression plasmid pBV220 to construct recombinant plasmid p pBV220/cTnI. The recombinant plasmid was transformed into E.coli DH5? which then expressed cTnI. The immunological activity of the expressed cTnI was analyzed by Western blot. Recombinant human cTnI protein was used as antigen to immunize BALB/c mice. Monoclonal anti-bodies against cTnI were prepared by normal hybridoma technology. The relative affinity of mAbs was determined by ELISA. Specificity of mAbs was analyzed by Western blot.Results Human cTnI gene was synthesized and confirmed by DNA sequencing. Positive recombinant clones were identified by restriction enzyme digestion analysis and DNA sequencing. Western blot analysis showed that the cTnI protein could be recognized by an anti- cTnI antibody. Two hybridmas producing antibodies against cTnI were obtained. IgG isotypes of two mAbs were IgG2a and IgG2b. Western blot showed that the antibodies were specific for cTnI. Neutralisation test showed that these mAbs could be evidently neutralized by cTnI.Conclusion The recombinant expression plasmid of cTnI was constructed successfully and expressed in E.coli. The method of EL ISA established to test serum cTnI is to clinically useful. The cTnI mAb which using cTnI as antigen prepared in this paper can be used for cTnI immunoassay in vitro.
2.The Early Diagnostic Value of Dynamic Changes for High-sensitive Troponin I in Patients With Acute Coronary Syndrome
Neng ZHANG ; Dan ZHANG ; Yu ZHANG ; Zhenyu YANG
Chinese Circulation Journal 2016;31(1):25-30
Objective: To explore the early diagnostic value of dynamic changes for high-sensitive troponin I (hs-cTnI) in patients with acute coronary syndrome (ACS).
Methods: A total of 186 ACS patients treated in our hospital from 2014-02 to 2015-04 were studied. The patients were divided into 2 groups:AMI group, n=169 and UA group, n=17, in addition, there was a Control group, n=13 healthy subjects. Blood levels of hs-cTnI were measured at admission and 2h, 4h after admission. Dynamic changes of hs-cTnI was calculated by slop coefifcient of hs-cTnI (Δhs-cTnI) at 3 time points and relative changes of hs-cTnI values. ROC curve was made by SPSS 16.0 statistical software and the best diagnostic cutoff point was determined by Youden index.
Results: In clinical practice, hs-cTnI=1 ng/ml was critical condition, hs-cTnI=0.15 ng/ml was recommended for AMI diagnosis. When hs-cTnI<1 ng/ml at admission, the AUC of AMI vs UA and AMI vs Control for 0 h hs-cTnI were 0.6746 and 0.8844;for relative changes of hs-cTnI were 0.9806 and 0.9631;forΔhs-cTnI were 0.9521 and 0.9778 respectively. Thus, the best diagnostic cutoff value of AMI for relative changes of hs-cTnI was 0.6705, sensitivity 70.59%, speciifcity 91.49%;forΔhs-cTnI was 0.0075, sensitivity 68.75%, speciifcity 97.78%. When hs-cTnI<0.15 ng/ml at admission, the AUC of AMI vs UA and AMI vs Control for 0 h hs-cTnI were 0.5677 and 0.7000;for relative changes of hs-cTnI were 0.9228 and 0.9975;forΔhs-cTnI were 0.9044 and 0.8906 respectively. Thus, the best diagnostic cutoff value of AMI for relative changes for hs-cTnI was 2.923, sensitivity 81.25%, speciifcity 94.12%;forΔhs-cTnI was 0.01125, sensitivity 76.47%, speciifcity 93.75%.
Conclusion: AMI could not be effectively diagnosed by hs-cTnI alone at admission, using combined relative changes of hs-cTnI andΔhs-cTnI was superior to 0 h hs-cTnI for AMI diagnosis.
3.Changes in biological behaviors of rat dermal fibroblasts induced by high expression of MMP9
Sheng-Neng XUE ; Juan LEI ; Diao-Zhu LIN ; Chuan YANG ; Li YAN
World Journal of Emergency Medicine 2014;5(2):139-143
BACKGROUND:The high level of matrix metalloproteinase 9 (MMP9) is thought to slow down the healing of diabetic foot ulcers. Whether it can influence the biological behaviors of skin fibroblasts and affect wound healing is stillunclear. The present study aimed to observe changes in the biological behaviors of rat dermal fibroblasts induced by high expression of MMP9 and to clarify the possible mechanisms of wound healing for diabetic foot. METHODS:A cellmodel of skin fibroblast with high expression of MMP9 was established by co-culture of high glucose (22.0 mmol/L) and homocysteine (100 μmol/L). A control group was incubated with normal glucose (5.5 mmol/L). Realtime PCR, ELISA and gelatin zymography were used to detect the MMP9 mRNA, protein expression and activity of MMP9. Flow cytometry, CCK-8, ELISA assay, scratch test and transwellwere used to detect cellproliferation, viability, collagen (hydroxyproline) secretion, horizontal migration and vertical migration of cells. The data were expressed as mean±SD. P value less than 0.05 was considered statistically significant. RESULTS:The expression of MMP9 mRNA, protein levels and the activity of MMP9 were much higher in the high MMP9 group than in the control group (7.05±1.02 vs. 1.00±0.00, 206.9±33.6 pg/mL vs. 40.4±5.9 pg/mL, and 1.47±0.13 vs. 0.57±0.12, respectively,P<0.01). The proportion of S-phase cells, proliferation index, cellviability, collagen (hydroxyproline) secretion, horizontal migration rate and the number of vertical migration cells were lower in the high MMP9 group than in the control group (P<0.01). CONCLUSION:Fibroblasts with a high expression of MMP9 decreased proliferation, activity, secretion and migration of collagens, suggesting that MMP9 may inhibit the biological behaviors of fibroblasts.
4.Applying of Zhang-Lu Index in Evaluating Physical Growth of Neonates in Huaihua City
li-lan, YI ; qing, LU ; jin-hua, YANG ; shi-neng, ZHAO
Journal of Applied Clinical Pediatrics 2006;0(21):-
Objective To validate the practical value of zhang-lu index(ZLI) in evaluating the physical growth of neonates.Methods Collecting the data of body weight,height and head circumference from 413 neonates,the results of ZLI were calculated according to the ZLI expression,then through the original date coming from 31 small for gestational age(SGA),65 large for gestational age(LGA) and 317 appropriate for gestational age(AGA),ZLI was compared with the conventional method of body weight for gestational age.Results 1.Compared with the method of body weight for gestational age,the sensitivity was higher than the result validated by the nation.The diffe-(rence) was significantly(P0.05).2.The results of ZLI of neonates in Huaihua city were larger than the reference given by the national of neonatal scientific research associate;The differences were significantly(P
5.A case-control study on risk factors for nosocomial infection of epilepsy in hospitalized children.
Neng-feng XU ; Liu-qing YANG ; Yong-quan HUANG
Chinese Journal of Epidemiology 2003;24(9):827-830
OBJECTIVETo determine the incidence and risk factors for nosocomial infection in child epilepsy.
METHODSA retrospective study was conducted among all (292 cases) hospitalized children epilepsy patients in First Affiliated Hospital of Fujian Medical University from 1996 to 2000 in Fuzhou city. With all patients with nosocomial infection as cases and all patients without nosocomial infection as controls, a case-control study on risk factors for nosocomial infection was carried out. Available data were analyzed by one-way Chi-square test and unconditional logistic multiple regression model.
RESULTSOne hundred fourteen cases of nosocomial infection were identified among 292 cases with epilepsy with an incidence of 39.0% (114/292). The one-way Chi-square test showed that nosocomial infection was significantly associated with age below 3 years (OR = 2.55, P < 0.01), length of hospitalization over 14 days (OR = 4.75, P < 0.01), low intelligence (OR = 3.13, P < 0.01), receiving antibiotic unreasonably (OR = 3.51, P < 0.01), using gastrogarage (OR = 3.12, P < 0.01), other invasive operation (OR = 1.85, P < 0.05) dyskinesia or palsy (OR = 3.51, P < 0.01), and urinary nitrogen beyond normal range (OR = 5.00, P < 0.05), etc. Unconditional logistic multiple regression analysis revealed that the length of hospitalization over 14 days (OR = 4.30, OR 95% CI: 2.48 - 7.46, P < 0.01), taking antibiotic unreasonably (OR = 2.74, OR 95% CI: 1.30 - 5.77, P < 0.01), using gastrogarage (OR = 3.04, OR 95% CI: 1.28 - 7.18, P < 0.05), and low intelligence (OR = 2.32, OR 95% CI: 1.34 - 4.01, P < 0.01) were independent risk factors for nosocomial infection. The tendency chi-square test showed that the longer stay in the hospital with more kinds of antibiotic used and more gastrogarage they used, the greater the risk of nosocomial infection was.
CONCLUSIONData suggested that occurrence of the nosocomial infection of children epilepsy patients was correlated with the length of hospitalization over 14 days, unreasonable using antibiotic, using gastrogarage and low intelligence.
Case-Control Studies ; Child ; Child, Hospitalized ; Child, Preschool ; Cross Infection ; etiology ; Epilepsy ; complications ; Female ; Humans ; Infant ; Length of Stay ; Logistic Models ; Male ; Retrospective Studies ; Risk Factors
6.Effect of casein and protamine on the enzymatic degradation and the orally hypoglycemic action of insulin.
Rong QI ; Qi-neng PING ; Rui-yang XU ; Yong-ping SHI
Acta Pharmaceutica Sinica 2004;39(10):844-848
AIMTo study the protection of casein and protamine against degradation of insulin (INS) by proteolysis enzymes and the effect of these two kinds of protein on the hypoglycemic action of INS solution and enteric-microspheres after administrated orally to rats.
METHODSHPLC was used to determine the remained INS in the solution of alpha-chymotrypsin and trypsin with or without casein or protamine; INS solution and enteric-microspheres were prepared and adiministrated orally to rats together with the absorption enhancer sodium N-[8-(2-hydroxybenzoyl) amino] caprylate (SNAC). At the same time, casein or protamine or both of these two kinds of protein were administrated together in order to study their influence on the hypoglycemic effect of INS and microspheres.
RESULTSCasein had a good protection against degradation of INS by alpha-chymotrypsin, but protamine had no protection effect. However, the degradation of INS by trypsin is concerned, the protection effect of protamine on INS was better that of casein. Both of protamine and casein can increase the hypoglycemic effect of INS solution and enteric-microspheres. Co-administrated these two kinds of protein had a better effect. In addition, co-administrated with SNAC, casein and protamine, INS enteric-microspheres had a longer and more potent hypoglycemic effect than that of the solution.
CONCLUSIONCasein and protamine can increase the stability of INS in the intestinal fluid by the mechanism of competition and combine with proteolysis enzymes, which will benefit to INS oral administration.
Administration, Oral ; Animals ; Blood Glucose ; metabolism ; Caprylates ; Caseins ; pharmacology ; Chymotrypsin ; antagonists & inhibitors ; Drug Delivery Systems ; Hypoglycemic Agents ; administration & dosage ; pharmacokinetics ; Insulin ; administration & dosage ; pharmacokinetics ; Male ; Microspheres ; Protamines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Solutions ; Trypsin ; pharmacology
7.Water in oil microemulsions for transdermal delivery of fluorouracil.
Fang LIU ; Yan-yu XIAO ; Qi-neng PING ; Chuang YANG
Acta Pharmaceutica Sinica 2009;44(5):540-547
An Aersol-OT (AOT) included microemulsion containing fluorouracil was prepared by using appropriate proportion of oil, co-surfactant and water for increasing the drug transdermal delivery ability. According to the area of microemulsion basing on the pseudo-tertiary phase diagrams, the optimum formulation was screened initially. And the permeation flux of fluorouracil across excised mice skin was determined in vitro using Franz diffusion cell to optimize the formulation further. The effect of the kind of co-surfactant, the content of water, the content of mixed surfactant, the mass ratio of surfactant/cosurfactant (Km) and the drug load on skin permeation of fluorouracil were evaluated. The optimum formulation was composed of 0.5% (w/v) fluorouracil, 30% water, 20% mix-surfactant (AOT/Tween 85, Km = 2) and 49.5% oil (IPM). The cumulative amount permeated of fluorouracil in 12 hour was 1 355.5 microg x cm(-2), 19.1 folds and 7 folds more than 0.5% fluorouracil aqueous solution and 2.5% (w/w) fluorouracil cream, respectively. The permeation of this microemulsion accorded with first-order model. The water/AOT/Tween 85/IPM microemulsion system promoted the permeation of fluorouracil greatly, which may be a promising vehicle for the transdermal delivery of fluorouracil and other hydrophilic drug.
Administration, Cutaneous
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Animals
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Antimetabolites, Antineoplastic
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administration & dosage
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pharmacokinetics
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Drug Carriers
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Drug Delivery Systems
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methods
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Emulsions
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Fluorouracil
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administration & dosage
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pharmacokinetics
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Male
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Mice
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Myristates
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chemistry
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Oils
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chemistry
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Polysorbates
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chemistry
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Skin Absorption
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Succinates
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chemistry
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Surface-Active Agents
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chemistry
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Water
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chemistry
8.Chemical constituents of Ligularia duciformis.
Kai-Zhou FU ; Neng-Jiang YU ; Yang ZHANG ; Yi-Min ZHAO
Acta Pharmaceutica Sinica 2007;42(6):621-624
Ligularia duciformis is a plant of the Ligularia genus (Compositae) which has been reported to have therapeutic effects of treating diseases such as bronchitis and coughing. In order to find out the bioactive components, the 85% ethanol extract of the root and rhizome of L. duciformis was treated by solvents extraction and column chromatography on silica gel and Sephadex LH-20. Six compounds were obtained from the ethyl acetate fraction and identified as 3-methoxy-4-hydroxyphenylpropyl caffeate (I), 6beta, 8beta-dihydroxyeremophil-7 (11)-en-12, 8alpha-olide (II), 6beta, 8alpha-dihydroxyeremophil-7 (11)-en-12, 8beta-olide (III), 6beta-hydroxyeremophil-7 (11)-en-12, 8alpha-olide (IV), stigmasterol (V) and beta-daucosterol (VI) by analysis of their physic-chemical constants and spectral data. Compound I is a new compound and compounds II - V were isolated from L. duciformis for the first time. The antitussive effect of the compounds is under evaluation.
Asteraceae
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chemistry
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Magnetic Resonance Spectroscopy
9.Effects of Ca2+ on photosynthetic parameters of Pinellia ternata and accumulations of active components in heat stress.
Wei-Xing YANG ; Gang-Gang HEI ; Jiao-Jiao LI ; Hong-Min ZHANG ; Lin-Lin LI ; Neng-Biao WU
China Journal of Chinese Materia Medica 2014;39(14):2614-2618
OBJECTIVETo study the effect of exogenous Ca2+ on photosynthetic parameters of Pinellia ternate and accumulations of active components under high temperature stress.
METHODThe pigment contents of P. ternata leaves, photosynthesis parameters and chlorophyll fluorescence parameters of P. ternata leaves, the contents of guanosine, adenosine and polysaccharide in P. ternata tubers were measured based on different concentrations of exogenous Ca2+ in heat stress when the plant height of P. ternata was around 10 cm.
RESULTThe contents of total chlorophyll and ratio of chlorophyll a/b were relatively higher by spaying Ca2+. Compared with the control, spaying 6 mmol x L(-1) Ca2+ significantly enhanced the net photosynthetic rate (Pn), transpiration (Tr) and stomatal limitation (L8), but reduced intercellular CO2 concentration (C) in P. ternata leaves. With the increase of Ca2+ concentration, maximal PS II efficiency (Fv/Fm), actual photosynthetic efficiency (Yield) and photochemical quenching coefficient (qP) initially increased and then decreased, however, minimal fluorescence (Fo) and non-photochemical quenching coefficient (NPQ) went down first and then went up. The contents of guanosine and polysaccharide and dry weight of P. ternata tubers showed a tendency of increase after decrease, and the content of adenosine increased with the increase of Ca2+ concentration. The content of guanosine and polysaccharide in P. ternata tubers and its dry weight reached maximum when spaying 6 mmol x L(-1) Ca2+.
CONCLUSIONWith the treatment of calcium ion, the inhibition of photosynthesis and the damage of PS II system were relieved in heat stress, which increased the production of P. ternata tubers.
Breeding ; Calcium ; pharmacology ; Chlorophyll ; metabolism ; Dose-Response Relationship, Drug ; Heat-Shock Response ; drug effects ; Organ Size ; drug effects ; Photosynthesis ; drug effects ; Pinellia ; drug effects ; growth & development ; metabolism ; physiology ; Plant Leaves ; drug effects ; growth & development ; metabolism
10.Comparison study of clinical effect and complications between subfascial and submammary breast augmentation.
Yan-Qing YANG ; Neng-Qiang GUO ; Jia-Ming SUN ; Hong-Bo CHEN ; Hang MA ; Qiang LI
Chinese Journal of Plastic Surgery 2013;29(1):12-14
OBJECTIVETo compare the clinical effect and complications of subfascial breast augmentation and submammary breast augmentation.
METHODFrom Sept. 2009 to May 2012 , 25 patients with subfascial breast augmentation and 31 patients with submammary breast augmentation were observed. The postoperative results including visible implant edge or ripple, upper pole of the implant and long-term implant ptosis were compared respectively. The complications including hematoma, infection and capsular contraction were also recorded.
RESULTS56 cases were followed up for 2 months to 26 months. The incidence rate of visible implant edge or ripple was 4.0% (1/25 ) in the subfascial group and 29.0% (9/31) in the submammary group, showing a significant difference between them ( PC 0.05). The incidence rate of convex upper pole of the implant was 8.0% (2/25) in the subfascial group and 35.5% (11/31) in the submammary group, showing a significant difference between them ( P < 0.05). Long-term implant ptosis was not found in the two groups. The incidence rate of hematoma was 4.0% (1/25) in the subfascial group and 6.5% (2/31) in the submammary group, infection was not found. The incidence rate of capsular contraction was 8.0% (2/25) in the subfascial group and 12.9% (4/31) in the submammary group, showing no statistical difference between them ( P > 0.05 ).
CONCLUSIONSSubfascial breast augmentation has more clinical advantages compared with submammary breast augmentation, but no evident difference was found in the common complication rate, such as capsular contraction.
Adult ; Female ; Humans ; Mammaplasty ; adverse effects ; methods ; Middle Aged ; Postoperative Complications ; Treatment Outcome ; Young Adult