1.Effects of human annexin Ⅴ derivative on coagulation and thrombosis: Effective time and dosage
Junyou CUI ; Liansheng WANG ; Chengwei JU ; Xiang YANG ; Zichun HUA
Chinese Journal of Tissue Engineering Research 2005;9(27):192-194
BACKGROUND: As a common anticoagulant, heparin is widely used in clinic, but it has remarkable side effects such as severe bleeding and heparin-induced thrombocytopenia, and it cannot inactivate fibrin-bound thrombin. Annexin Ⅴ derivative (AND) is inosculated with C-terminal of hirudin and annexin Ⅴ, and its anticoagulation and anti-thrombosis effects are compared with those of heparin. OBJECTIVE: To investigate the relationship between quantitative effectiveness and time effectiveness of AND on coagulation and thrombosis, and study its reliability. DESIGN: Completely randomized grouping design and controlled study. SETTING: Cardiac Department of amunicipal hospital. MATERIALS: The experiment was conducted at the Animal Laboratory of Jiangsu Provincial People's Hospital from July 2000 to April 2001. Totally 32 male New Zealand white rabbits were randomly divided into 4groups, namely, high dosage AND group, low dosage AND group, common heparin group and saline group with 8 in each group. METHODS: Heparin and AND were diluted with saline.①High dosage AND group: 0.7 mg/kg AND was injected intravenously and followed by intravenous dripping of 0.35mg/(kg ·h)for 2 hours.Low dosage AND group: 0.3 mg/kg AND was injected intravenously and followed by intravenous dripping of 0.15 mg/(kg·h) for 2 hours. Heparin group: 75 IU/kg heparin was injected intravenously and followed by intravenous dripping of 37.5 IU/(kg·h) for 2 hours. Saline group: The same volume of saline and medication were used as those in drug groups.② Blood sample was collected from the femoral vein before administration so as to test blood routine, activated partial thromboplastin time(APTT)and prothrombin time (PT) after 15-, 30- and 60-minute administration and 2-hour withdrawal.③Saccule was separated from endothelium of femoral artery to measure blood pressure of distal femoral artery at 15 minutes after administration.Time of pulse pressure equal to 0 mmHg was recorded when the vessel was occluded completely by a thrombus.Finally the injured femoral arteries whose vessel was stripped were collected to measure its length, wet weight and dry weight. ④Observation of AND toxicity and sideeffects:During the experiment,vital signs of the animals were measured,such as blood pressure,heart rate and breath;in addition,bowelhemorrhage was observed and the number of leucocytes was counted after dissection of some of the animals. MAIN OUTCOME MEASURES:①Effect of AND on blood coagulation system and arterial thrombosis.②AND toxicity and side effects. RESULTS: All the 32 white rabbits entered the final analysis. ① Anticoagulant effect: APTT: Fifteen minutes after administration, APTT in AND group was the longest,which was(136.86±39.46)s in high dosage AND group and (122.90±34.19) s in low dosage ANDgroup.Moreover, APTT was longer than that in saline group [(95.14±24.64) s], but shorter than that in common heparin group [(180.00±0.00) s, P < 0.05, 0.01]. At 30 minutes after administration,AND in high dosage group still had coagulation,and APTT was (124.61±40.19) s in high dosage group, which was longer than that in saline group [(85.57±27.67) s], but APTT was (112.94±43.17) sin low dosage group,which was shorter than that in common heparin group [(85.57±27.67)s,P < 0.05].APTT was shorter in high and low dosage groups than in common heparin group at 60 minutes after administration (P < 0.05),and longer than that in saline group 2 hours after drug withdrawal,but there was not significant difference(P > 0.05).PT:PT in common heparin group was longer than that in high and low dosage groups at 15,30 and 60 minutes after administration (P < 0.05).② Effect on arterial thrombosis:Wet weight of thrombus:It was lighter in AND group than in common heparin group(P < 0.05). Dry weight of thrombus:Thrombus was lighter in high and low dosage groups than in common heparin group, and was lighter in high dosage group than in low dosage group (P < 0.05).Thrombus length:It was shorter in low dosage group than in saline group (P < 0.05), and shorter in high dosage groupthan in common heparin group (P < 0.05). Time of complete occlusion: It was longer in high and low dosage groups than in saline group(P < 0.05).③ AND toxicity and side effects:The behavior of rabbits in high and low dosage groups was similar to that in other two groups. Obvious hemodynamic changes were not found, and bowel hemorrhage was not observed, either. CONCLUSION: AND is an effective anticoagulant and anti-thrombosis agent; the highest anticoagulation effect occurs at 15 minutes afteradminis tration. However, the anticoagulant effect is poor as compared to heparin.The effect is poorer after 60-minute administration. Effect of AND on thrombus is stronger than that of heparin,but the size of thrombus is smaller than that of heparin, and the dosage-dependence manner was found. In addition, the anti-thrombus effect of AND is stronger in high dosage group than in low dosage group.
2.Experimental study of cell transplantation into ischemic myocardium using mesenchymal stem cells transfected by vascular endothelial growth factor
Ping HUA ; Ju CHEN ; Hui-Zhong ZHANG ; Song-Ran YANG ; Yan-Qi YANG ; Li-Hua XIONG ; Hua ZHANG ;
Chinese Journal of Microsurgery 2006;0(05):-
Objective To investigate the effects of mesenchymal stem cells (MSCs) transplantation combining with vascular endothelial growth factor (VEGF) gene therapy on myocardium rebuilding,angiogene- sis,and heart function improvement in rats with myocardial infarction.Methods SD rat MSCs were isola- ted,cultured in vitro,labeled with BrdU and transfected by Ad.VEGF gene.Four weeks after left anterior descending artery was ligated to created rat myocardial infarction,cardiac function was examined with echocar- diography,rats were randomly divided into four groups (n=10 in each group):GroupⅠ:MSCs/Ad.VEGF implantation;GroupⅡ:MSCs implantation;GroupⅢ:Ad.VEGF injection;GroupⅣ:Control.MSCs dif- ferentiation was observed 4 weeks after transplantation.Immunohistochemistry and angiogenesis were observed. Echocardiography was performed to detect the effects on heart function.Results MSCs labeled with BrdU could be identified in host hearts in groupⅠandⅡ,most of them positively stained with cTnT antibody. Echocardiography indicated that the improvement of the LVEF value in groupⅠwas more significant than that in the other three groups (P<0.01,respectively).Some cells were incorporated into the coronary capillaries in the infarcted region.The capillary density in groupⅠwas higher than that in the other three groups (P<0.01,respectively).Conclusion MSCs implantation combining with VEGF gene therapy can obviously re- pair damaged myocardium and enhance the angiogenesis in ischemic heart tissue.
3.Qianlongtong capsule elevates the Smad4 gene expression in prostate stromal cells.
Zhi LING ; Hui LIU ; Jing YANG ; Ju-Qiao HE ; Sai YANG ; Qi-Hua CHEN
National Journal of Andrology 2014;20(8):730-733
OBJECTIVETo investigate the effects of the plasma containing Qianlongtong Capsule (QLT)-containing plasma on the expression of the Smad4 gene in prostate stromal cells in vitro and provide some experimental evidence for the treatment of benign prostatic hyperplasia (BPH) with Chinese medicinal compound.
METHODSFifteen cases of BPH were equally randomized to three groups to be treated with QLT at a high dose (6 capsules once), a medium dose (3 capsules once), and a low dose (1.5 capsules once), tid, for 7 days consecutively. QLT-containing plasma was collected from the patients. Prostate stromal cells were identified by immunofluorescence when they became monolayered and cultured in the QLT-containing plasma for 24 hours, followed by detection of the expression of the Smad4 gene by real-time quantitative PCR and that of the Smad4 protein by Western blot.
RESULTSAfter treatment with the QLT-containing plasma, the expression of the Smad4 gene in the stromal cells was significantly increased in a dose-dependent manner as compared with the blank control and no-QLT groups (P < 0.01). The expression of the Smad4 protein was also markedly elevated after treatment. The differences were statistically significant between the blank control and medium-dose groups (P < 0.01), low-dose and medium-dose groups (P < 0.05), and high-dose and the other groups (P < 0.01), but not between the blank control and low-dose groups (P > 0.05).
CONCLUSIONQLT-containing plasma could inhibit the proliferation and improve the apoptotic index of prostate stromal cells in vitro, which was related to the elevation of the mRNA and protein expressions of Smad4.
Apoptosis ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Male ; Prostate ; drug effects ; metabolism ; RNA, Messenger ; genetics ; Smad4 Protein ; genetics ; metabolism ; Stromal Cells ; drug effects ; metabolism
4.Liquisolid technique for enhancement of dissolution prosperities of tanshinone II(A).
Xiao-qian LIU ; Qing-ju MENG ; Xue-lin XU ; Jie ZHAO ; Hua YANG ; Hong YI
China Journal of Chinese Materia Medica 2015;40(24):4840-4846
The technique of liquisolid compress is a new technique developed in 1990s, which was considered to be the most promising technique to improve the dissolution of water-insoluble drugs. In this article, tanshinone II(A) and the extracts of the ester-solubility fractions were chosen as the model drugs to evaluate the effects of the liquisolid technique for enhancement of dissolution properties of tanshinone II(A). Several liquisolid tablets (LS) formulations containing different dosage of drugs and various liquid vehicle were pre-pared and for all the formulations, microcrystalline cellulose and silica were chosen as the carrier and coating materials to evaluate their flow properties, such as angle of repose, Carr's compressibility index and Hausner's ratio. The interaction between drug and excipients in prepared LS compacts were studied by differential scanning calorimetry(DSC) and X-ray powder diffraction (XRPD). The dissolution curves of tanshinone II(A) from liquisolid compacts were investigated to determine the technique's effect in improving the dissolution of tanshinone II(A) and its impacting factors. According to the results, the dissolution increased with the rise in the dissolution of the liquid-phase solvent. The R-value and drug dosage can significantly affect the drug release, but with less impact on active fractions. This indicated that liquisolid technique is a promising alternative for improvement of dissolution property of water-soluble drugs, and can make a synergistic effect with other ester-soluble constituents and bettern improve the release of tanshinone II(A). Therefore, the technique of liquisolid compress will have a better development prospect in traditional Chinese medicines.
Calorimetry, Differential Scanning
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Diterpenes, Abietane
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chemistry
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Solubility
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X-Ray Diffraction
5.Clinical Study of Relationship between Free Thyroxine Levels and Serum Tumor Necrosis Factor, Albumin, Urinary Protein in Children with Nephrotic Syndrome
qiaozhi, YANG ; xueyun, LV ; ai-ju, YU ; ai-hua, CUI ; jun, LIANG
Journal of Applied Clinical Pediatrics 1992;0(05):-
Objective To study the changes of serum free triiodothyronine (FT3), free tetraiodothyronine (FT4) and the relation between free thyroxine levels and serum tumor necrosis factor (TNK), albumin (ALB), urinary protein in children with primary nephrotic syndrome(PNS). Methods There were sixty children who were suffered from nephrotic syndrome in study group Serum FT3,FT4,TNF, Alb and urinary protein were detected. In the meantime compared with 25 health,cases. Results The levels of FT3, FT4 of the children who were suffered from nephrotic syndrome were lower. The difference between nephrotic syndrome and health cases were significantly (P
6.Echocardiography diagnose one case of infantile tricuspid myxoma.
Chuan-ju HOU ; Yu-hua CAO ; Dong-an DENG ; Xian-yang ZHU
Chinese Journal of Pediatrics 2004;42(7):553-553
Echocardiography
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methods
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Female
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Heart Neoplasms
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diagnosis
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surgery
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Humans
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Infant
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Myxoma
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diagnosis
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surgery
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Treatment Outcome
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Tricuspid Valve
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physiopathology
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surgery
7.Preparation and release behaviour of mPEG-PLA α-asarone nanoparticles designed for nasal administration.
Feng JU ; Lin-mei PAN ; Li-wei GUO ; Hua-xu ZHU ; Bo LI ; Na QU ; Chen YANG
China Journal of Chinese Materia Medica 2015;40(24):4847-4852
Taking α-asarone as model drug, mono methoxy polyethylene glycol-polylactic acid copolymer (mPEG-PLA) as the drug carrier material to prepare drug-loading nanoparticles by premix membrane emulsification for nasal administration. The prepared nanoparticles were spherical with smooth surface and average particle size of 360 nm. Polydispersity index (PDI) was 0. 030, average drug loading of (11.5 ± 0.045) % (n = 3), and the encapsulation efficiency of (86.34 ± 0.11) % (n = 3). X-ray diffraction and differential scanning calorimetry results showed that, α-asarone existed in mPEG-PLA carrier in amorphous or molecular state, different from simple physical mixture. In the in vitro release test in simulated human nasal cavity, α-asarone apis can be released quickly at close to 94% at 102 h, in line with the first-order kinetics (R² = 0.981 9). mPEG-PLA drug-loading nanoparticles release only 54%, with slow release effect, in line with Riger-Peppas model (R² = 0.967 9, n = 0.630 2), for non-fick diffusion, released by the spread of drugs and skeleton dissolution dual control. This provided the foundation for nasal drug delivery in vivo pharmacokinetic study.
Administration, Intranasal
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Anisoles
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chemistry
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Calorimetry, Differential Scanning
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Nanoparticles
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chemistry
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Polyesters
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chemistry
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Polyethylene Glycols
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chemistry
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Solubility
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X-Ray Diffraction
8.Relationship of expression of extracellular matrix metalloproteinase inducer and hepatocyte growth factor with lymphoid metastasis and prognosis in non-small-cell lung carcinoma
Huizhong ZHANG ; Mei WANG ; Yiping WEI ; Yongke CAO ; Haigang LI ; Ju CHEN ; Yanqi YANG ; Ping HUA ; Cheng WU ; Jingeng LIU
Chinese Journal of Pathophysiology 1999;0(09):-
AIM:To investigate the expression of extracellular matrix metalloproteinase inducer(EMMPRIN)and hepatocyte growth factor(HGF)in non-small-cell lung carcinoma(NSCLC)and their relationship with lymphoid metastasis and prognosis.METHODS:Expression of EMMPRIN and HGF in 77 cases of patients with NSCLC was detected immunohistochemically.The relationship of expression of EMMPRIN and HGF with tumor size,smoking,histological type,differentiation,lymphoid metastasis,clinical stage,and prognosis was analyzed.RESULTS:The expressive rates of EMMPRIN and HGF were 68% and 44%,respectively.The expressions of EMMPRIN and HGF were associated positively with lymphoid metastasis(r=0.371 and 0.339,P0.05).The expression of EMMPRIN was associated with the expression of HGF in NSCLC.CONCLUSION:The expression of EMMPRIN and HGF is associated with lymphoid metastasis and prognosis in NSCLC.Overexpression of EMMPRIN and HGF implies infavourable prognosis in NSCLC.
9.Antiviral Effect of Interferon-Induced Guanylate Binding Protein-1 against Coxsackie Virus and Hepatitis B Virus B3 in Vitro
Yin-ping, LU ; Bao-ju, WANG ; Ji-hua, DONG ; Zhao, LIU ; Shi-he, GUAN ; Meng-ji, LU ; Dong-liang, YANG
Virologica Sinica 2007;22(3):193-198
Guanylate binding protein-1(GBP-1) is an interferon-induced protein. To observe its antiviral effect against Hepatitis B virus (HBV) and Coxsackie virus B3 (CVB3), we constructed an eukaryotic expression vector of human GBP-1(hGBP-1). Full-length encoding sequence of hGBP-1 was amplified by long chain RT-PCR and inserted into a pCR2.1 vector, then subcloned into a pCDNA3.1(-) vector. Recombinant hGBP-1 plasmids and pHBV1.3 carrying 1.3-fold genome of HBV were contransfected into HepG2 cells, and inhibition effect of hGBP-1 against HBV replication was observed. Hela cells transfected with recombinant hGBP-1 plasmids were challenged with CVB3, and viral yield in cultures were detected. The results indicated that recombinant eukaryotic expression plasmid of hGBP-1 was constructed successfully and the hGBP-1 gene carried in this plasmid could be efficiently expressed in HepG2 cells and Hela cells. hGBP-1 inhibit CVB3 but not HBV replication in vitro. These results demonstrate that hGBP-1 mediates an antiviral effect against CVB3 but not HBV and perhaps plays an important role in the interferon-mediated antiviral response against CVB3.
10.Construction and Characterization of a Hepatitis B Virus Replicon
Yin-ping, LU ; Bao-ju, WANG ; Ji-hua, DONG ; Zhao, LIU ; Shi-he, GUAN ; Meng-ji, LU ; Dong-liang, YANG
Virologica Sinica 2007;22(1):8-13
To establish a replication cellular model of hepatitis B virus (HBV) and determine its application in antiviral drug evaluation,we constructed an expression plasmid which contained 1.3 copies of the HBV genome,and measured the level of viral replication after transient transfection in Huh7 cells.We then observed the effect of antiviral drug administration.1.3 fold of the HBV(ayw) gene fragment was cloned into pCR2.1 by PCR and restriction endonuclease digestion.The recombinant plasmid was trans ient transfected into Huh7 cells,HBsAg,HBeAg and HBV DNA in supernatant of Huh7 cells were measured by ELISA and real-time PCR respectively; intracellular HBV replicative intermediates and intracellular HBV transcripts were detected by Southern blot and Northern blot respectively.The antiviral effect of adefovir,a novel anti-HBV nucleotide analogue,was evaluated in this cellular model system.The results indicated that a recombinant plasmid of HBV replicon was constructed successfully; the HBV genome carried in plasmid pHBV1.3 could efficiently replicate and be expressed in Huh 7 cells,adefovir could inhibit HBV replication in this cellular model,and the inhibition was dosage-dependent.The conclusion is HBV replicon,which can initiate viral replication efficiently in hepatoma cells,may be a useful tool in the study of HBV replication and antiviral drug.