1.Electroanalytical method for TPPS4 ,the interaction of TPPS4 with BSA and the influence of CDs on it by fluorescence spectroscopy
Hongfen ZHANG ; Jinghao PAN ; Haibo CHANG ; Yun LIU ; Yujing GUO ; Fei LU
Acta Pharmaceutica Sinica 2006;41(3):203-209
Aim To establish a simple, rapid and accurate electroanalytical method for water soluble porphyrin meso-tetrakis-(4-sulfonatophenyl) porphyrin (TPPS4);to clarify the reaction between water soluble porphyrins and bovine serum albumin (BSA);and to determine the interaction of TPPS4 with BSA in the absence of presence of cyclodextrins (CDs) , separately. Methods Three methods including LSV,UV spectroscopy and fluorescence spectroscopy bad been employed to the relevant experiments. The way of employing three methods at the same time could make the experiment results more reliable. Results In the supporting electrolyte of NaH2 PO4-Na2 HPO4 ( pH 7. 18 ), a sensitive reduction peak of TPPS4 was found by linear sweep voltammetry (LSV), the peak potential (Ep) was-0. 70 V (vs SCE). The relationship between the second derivative peak of LSV (ip") and the concentration of TPPS4 was linear from 1.0 × 10-7mol·L-1 to 1.0 × 10-5mol· L-1,the square of correlation coefficients (r2) were 0. 998 3 and 0. 999 3, respectively. The relative standard deviation (RSD) was 0. 56% ( n = 5 ). The mean recovery of TPPS4 was 99.59%. In NH4C1-NH3· H2O buffers (pH 9.05), it was proved that BSA and TPPS4 could interact with each other and form 1:1 TPPS4-BSA supramolecular system. Moreover, the interaction between TPPS4 and BSA had been investigated by adding cyclodextrins (CDs). The interaction of TPPS4 with BSA was facilitated both by hydroxypropyl-β-CD (HP-3-CD) and sulforbutylether-β-CD (SBE-3-CD). Conclusion An electroanalytical method for TPPS4 has been established by LSV. The porphyrin drugs included by CDs could react with protein existing inside the human body easier. The consequences of this article also show that CDs will play important role in controlling and releasing the porphyrin drugs.
2.Expression of C2A domain of synaptotagmin Ⅰ fusion protein and its imaging in the ischemia-reperfusion rat model
Kang-yun, SUN ; Jun-dong, ZHOU ; Fei, SHEN ; Wei, FANG ; Jin-chang, WU
Chinese Journal of Nuclear Medicine 2010;30(5):312-315
Objective To evaluate myocardial apoptosis with 99Tcm-C2A-GST myocardial imaging using the recombined C2A domain of Synaptotagmin Ⅰ by gene engineering. Methods ( 1 ) The C2A gene was inserted into the prokaryotic glutathione S-transferate (GST) fusion protein expression plasmid pGEX-6P-1. The recombinant plasmid was transformed into E. coli BL21. C2A-GST fusion protein was purified after BL21 was induced with isopropyl-β-D-1-thiogalactopyranoside (IPTG). (2)The activity of fusion protein was identified by cell binding test with fluorescein-5-isothiocyanate (FITC)-C2A-GST. (3) The C2A-GST fusion protein was labeled with 99Tcm using 2-iminothiophene hydrocoride method. Radiochemical purity was determined with thin layer chromatography. (4)99Tcm-C2A-GST (7.4 MBq) was injected to ischemia-reperfusion rat models through tail vein. The image was acquired with SPECT at 1 h after injection, and then hearts were removed, rinsed with saline and dyed with triphenyl tetrazolium coride (TTC). The ischemic myocardium was separated from the viable myocardium and was weighted. Its radioactivity was measured by gamma counting. The difference of uptake of radiotracer between ischemic myocardium and normal myocardium was compared using percentage activity of injected dose per gram of tissue ( % ID/g) with standard deviation. SPSS 12.0 and t-test were used for data analysis. Results ( 1 ) C2A-GST fusion protein wassuccessfully expressed and its relative molecular weight was 3.8 × 104. (2) FITC-C2A-GST binding to apoptotic cells could be observed by fluorescent microscopy. (3) The radiochemical purity of 99Tcm-C2A-GST was (98.90 ±0.43)%. (4)The imaging studies showed that there was focal uptake of radioactivity in the ischemic myocardium. In vitro uptake of 99Tcm-C2A-GST was (2.41 ±0.32) % ID/g by the ischemic myocardium, however 99Tcm-C2A-GST-N-hydroxysuccinimide (C2A-GST-NHS) was (0. 82 ± 0. 24) % ID/g. There was statistically significant difference between those two groups (t = 10. 6, P <0.01 ). Conclusion The C2A domain of Synaptotagmin Ⅰ expressed by gene engineering can be used as the tracer for noninvasive detection of ischemic myocardium in the ischemia-reperfusion rat model.
3.Relationship between three thrombophilic gene mutations and unexplained recurrent early spontaneous abortion
Li XU ; Xiao-Man LIU ; Hong-Yan ZHANG ; Jing ZHAO ; Qi-Wei QI ; Yun-Fei CHANG ;
Chinese Journal of Obstetrics and Gynecology 2001;0(03):-
Objective To explore the relationship of methylenetetrahydrofolate reductase (MTHFR)gene C677T,factor V(FV)gene G1691A and prothrombin(PT)gene G20210A polymorphisms to unexplained recurrent early spontaneous abortion(URESA).Methods One hundred and twelve patients with URESA and 100 women with at least 1 normal pregnancy and without any miscarriage were analyzed for MTHFR,FV and PT gene polymorphisms by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP).Results MTHFR gene T/T genotype and T allele frequencies were increased in URESA patients[38.4%(43/112)and 59.8%(134/224)]versus controls[18.0%(18/100)and 43%(43/100),P
4.An experimental study of the protective effects of the ultrashort wave diathermy on gentamycin-induced acute renal iujury
Xiang-Yun LI ; Zao-Ming HUANG ; Chang-Fie ZHANG ; Shi-Fei ZHANG ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(07):-
Objective To observe the treatment effects of the ultrashort wave diathermy on gentamycin-in- duced acute renal injury of rats.Methods Eighteen Sgrague-Dawley rats were randomly divided into a normal group(6 rats),a model group(6 rats)and a treatment group(6 rats),the treatment group was treated with ultra- short wave diathermy once a day for a total of 20 days.The observed indexes were NAG,RBP,?_1-MG,?_2-MG in the urine of rats;the SCr,BUN in the blood of rats were also tested and the pathological changes of the renal ob- served.Results The pathological injuries of those in the ultrashort wave treatment group was slighter than the con- trol group;the NAG,RBP,?_1-MG,?_1-MG in the urine and the SCr,BUN in the blood of treatment group were low- er than those of the model group(P
5.Alteration of cytokines and mucin in lower respiratory tract in allergic rhinitis model in rats.
Fei HAN ; Yun-Fang AN ; Chang-Qing ZHAO
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2005;40(5):339-342
OBJECTIVETo investigate the relationship between allergic rhinitis (AR) and asthma as well as the mechanisms related with it.
METHODSSixty healthy rats were randomly divided into AR group and control group. AR model was established by intraperitoneal injection of ovalbumin (OVA) and nasal challenge with OVA. Nasal mucosa and lung tissue from both groups were stained with hematoxylin-eosin (HE), alcian-blue and periodic acid-schiff (AB-PAS), respectively. At the same time, the lung tissue was studied by transmission electron microscopy (TEM). Enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry were used to examine the level of interleukin-4 (IL-4) in bronchial alveolus lavage fluid (BALF) and the expression of intercellular adhesion molecule-1 (ICAM-1) and MUC5AC in nasal and lung tissue, respectively.
RESULTSInfiltration of inflammatory cells in nasal mucosa and lung tissue of AR model in rat was evident. Cilia destruction of bronchial epithelial cells of AR model was found. The level of IL4 in BALF of AR group (58.10 +/- 7.92) pg/ml was significant higher compared with that in control group (24.66 +/- 2.07) pg/ml. The expression of ICAM-1 (0.66 +/- 0.24) and MUC5AC (0.71 +/- 0.10) in lung tissue were both significantly higher than that in control group (0.23 +/- 0.02, 0.29 +/- 0.03).
CONCLUSIONSAllergic inflammation in nasal mucosa not only leads to changes in both histopathology and immunology, but also initiates the inflammation in lower respiratory tract mainly causing the change of cytokines and mucin.
Animals ; Bronchoalveolar Lavage Fluid ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-4 ; metabolism ; Mucin 5AC ; metabolism ; Mucins ; metabolism ; Nasal Mucosa ; metabolism ; pathology ; Rats ; Rhinitis, Allergic, Perennial ; chemically induced ; metabolism ; pathology
6.Squamous cell papilloma in interdental papilla: a case report.
Chang-chang YE ; Hai-yun HUANG ; Yuan GAO ; Ping HUANG ; Ya-fei WU
West China Journal of Stomatology 2009;27(2):235-236
Squamous cell papilloma is a kind of benign tumor from mucosa stratified squamous epithelium, which usually occurs in cheek, palate, lip and tongue. In this paper, a case of squamous cell papilloma occurred in interdental papilla was reported, and its pathogenesis, clinic features and treatment were discussed.
Epithelial Cells
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Epithelium
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Gingiva
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Humans
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Papilloma
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Tongue
7.Pestalotiolide A, a New Antiviral Phthalide Derivative from a Soft Coral-derived Fungus Pestalotiopsis sp..
Yan Lai JIA ; Fei Fei GUAN ; Jie MA ; Chang Yun WANG ; Chang Lun SHAO
Natural Product Sciences 2015;21(4):227-230
Chemical investigation of the fermentation broth of a Soft Coral-Derived fungus Pestalotiopsis sp., led to the isolation of a new phthalide derivative, pestalotiolide A (1), three known analogues (2, 3 and 4), along with 5'-O-acetyl uridine (5) first isolated as a natural product. The structure of the new compound (1) was established by comprehensive spectroscopic analysis and chemical methods. Compounds 1 - 4 possessed varying degrees of antiviral activities, which was reported for the first time. Compared to the positive control ribavirin (IC50 = 418.0 microM), pestalotiolide A (1) exhibited significant anti-EV71 activity in vitro, with an IC50 value of 27.7 microM. Furthermore, the preliminary structure-activity relationship of antiviral activities was also discussed.
Fermentation
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Fungi*
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Inhibitory Concentration 50
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Ribavirin
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Structure-Activity Relationship
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Uridine
8.Clinical research of angle-splitting ostectomy based on three dimensional computed tomography true-up technique.
Yan-feng ZHAO ; Yun-fei HAO ; Ping LU ; Xiao-nan ZHOU ; Chang-feng QU
West China Journal of Stomatology 2009;27(5):516-520
OBJECTIVETo illustrate the morphological changes of mandible after angle-splitting ostectomy.
METHODSFrom January 2006 to April 2008, 10 cases had undergone mandibular angle-splitting ostectomy to reduce the width of the lower face. For each patient, CT datum of mandible at three stages (preoperative, immediate postoperative, 6 months postoperative) were collected. By the application software of reverse engineering (Surfacer V9) and true-up and dissection techniques based on three-dimensional spiral computed tomography (3D-CT), operative efficacy and bone regeneration at the operation area of angle-splitting ostectomy were evaluated 6 months postoperative.
RESULTS1) Concavity could be seen at the angle-splitting ostectomy area 6 months postoperative, especially at the mandibular external oblique line region. Average cup depth was (3.64 +/- 1.67) mm by contrasted to preoperative. Diminution of bone volume was 55% +/- 9% for the local operative area 6 months postoperative. 2) Bone regeneration could be seen at the area that mandibular outer cortex had been removed. Compared with immediate postoperative, ratio of neoformative bone was 84.6% +/- 7.3% 6 months postoperative. The main region of bone regeneration was mandibular angle.
CONCLUSIONMandibular angle-splitting ostectomy is an effective technique for reducing the width of the lower face. Masseter muscular movement should be restricted postoperative to prevent hyperostosis at the angle area.
Adult ; Bone Regeneration ; Face ; Female ; Humans ; Mandible ; Masseter Muscle ; Osteotomy ; Tomography, X-Ray Computed
9.Genetic polymorphism of D6S477, D9S1118, D18S865, D19S400, and D20S161 short tandem repeat loci in Qingdao Han population.
Qi-wei QI ; Hong-yan ZHANG ; Yun-fei CHANG ; Li XU ; Shou-zhong MA
Chinese Journal of Medical Genetics 2005;22(4):473-475
OBJECTIVETo illuminate the preliminary genotype and allele frequency distribution of D6S477 and the other four short tandem repeat(STR) loci in Chinese Han population in Qingdao area and to probe the possibility of their genetic application.
METHODSTwo hundred ACD-blood specimens were collected from the unrelated individuals in Qingdao. The DNA samples were extracted with Chelex method and were amplified by polymerase chain reaction technique. The PCR products were analyzed by polyacrylamide gel electrophoresis and displayed using silver staining.
RESULTSThe authors obtained the allele frequency distribution and preliminary genotype of D6S477, D9S1118, D18S865, D19S400 and D20S161 STR loci. No deviation from Hardy-Weinberg equilibrium was observed in the five loci.
CONCLUSIONAll the five loci have higher chance of exclusion and discriminating power, and they will be useful markers for researches in genetics.
Asian Continental Ancestry Group ; genetics ; China ; Gene Frequency ; Genotype ; Humans ; Linkage Disequilibrium ; Microsatellite Repeats ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic
10.Electroanalytical method for TPPS4, the interaction of TPPS4 with BSA and the influence of CDs on it by fluorescence spectroscopy.
Hong-fen ZHANG ; Jing-hao PAN ; Hai-bo CHANG ; Yun LIU ; Yu-jing GUO ; Fei LU
Acta Pharmaceutica Sinica 2006;41(3):203-209
AIMTo establish a simple, rapid and accurate electroanalytical method for water soluble porphyrin meso-tetrakis-(4-sulfonatophenyl) porphyrin (TPPS4); to clarify the reaction between water soluble porphyrins and bovine serum albumin (BSA); and to determine the interaction of TPPS4 with BSA in the absence of presence of cyclodextrins (CDs), separately.
METHODSThree methods including LSV, UV spectroscopy and fluorescence spectroscopy had been employed to the relevant experiments. The way of employing three methods at the same time could make the experiment results more reliable.
RESULTSIn the supporting electrolyte of NaH2 PO4-Na2 HPO4 (pH 7.18), a sensitive reduction peak of TPPS4 was found by linear sweep voltammetry (LSV), the peak potential (Ep) was -0.70 V (vs SCE). The relationship between the second derivative peak of LSV (ip") and the concentration of TPPS4 was linear from 1.0 x 10(-7) mol x L(-1) to 1.0 x 10(-5) mol x L(-1), the square of correlation coefficients (r2) were 0.998 3 and 0.999 3, respectively. The relative standard deviation (RSD) was 0.56% (n = 5). The mean recovery of TPPS4 was 99.59%. In NH4Cl-NH3 x H2O buffers (pH 9.05), it was proved that BSA and TPPS4 could interact with each other and form 1 : 1 TPPS4-BSA supramolecular system. Moreover, the interaction between TPPS4 and BSA had been investigated by adding cyclodextrins (CDs). The interaction of TPPS4 with BSA was facilitated both by hydroxypropyl-beta-CD (HP-beta-CD) and sulforbutylether-beta-CD (SBE-beta-CD).
CONCLUSIONAn electroanalytical method for TPPS4 has been established by LSV. The porphyrin drugs included by CDs could react with protein existing inside the human body easier. The consequences of this article also show that CDs will play important role in controlling and releasing the porphyrin drugs.
2-Hydroxypropyl-beta-cyclodextrin ; Electrochemistry ; methods ; Electrodes ; Porphyrins ; chemistry ; metabolism ; Protein Binding ; Serum Albumin ; chemistry ; metabolism ; Spectrometry, Fluorescence ; Spectrophotometry, Ultraviolet ; beta-Cyclodextrins ; chemistry ; metabolism