2.Effects of traditional Chinese medicine colquhounia root tablet on the expression of tight junction protein ;claudin-2 and ZO-1 in bronchial epithelium tissue of rats with acute lung injury induced by oleic acid
Ping SHAO ; Xueli LI ; Jinyuan ZHU ; Huan DING ; Xigang MA ; Xiangyuan CAO
Chinese Critical Care Medicine 2016;28(6):523-527
Objective To investigate the effects of traditional Chinese medicine colquhounia root tablet on the expression of tight junction protein claudin-2 and ZO-1 in bronchial epithelium tissues of rats with acute lung injury (ALI), and to study the mechanism of protective effect of colquhounia root tablet on ALI. Methods Twenty-four healthy male Sprague-Dawley (SD) rats were randomly divided into control group, ALI group and colquhounia root tablet pretreatment group, with 8 rats in each group. The model of ALI was reproduced by intravenous injection of oleic acid 0.04 mL/kg, and the rats in cont rol group were given the same amount of normal saline (NS) instead. The rats in colquhounia root tablet pretreatment group were intragastric administrated with colquhounia root tablet of 600 mg·kg-1·d-1 (2 mL) for 10 days before model reproduction, and the rats in control group and ALI group were given the same amount of NS. At 4 hours after model reproduction, the blood was drawn from abdominal aorta, and bronchoalveolar lavage fluid (BALF) was collected for determination of protein content in plasma and BALF, and the lung permeability index (LPI) was calculated. The rats were sacrificed to collect lung tissues for determination of lung wet/dry weight ratio (W/D), the changes in pathology of lung tissue were observed after hematoxylin and eosin (HE) staining with light microscope, and lung injury score (LIS) was evaluated. The immunohistochemic al staining was used to detect the expression and localization of claudin-2 and ZO-1 in bronchial epithelium tissues. The protein expressions of claudin-2 and ZO-1 in bronchial epithelium tissues were determined by Western Blot. Results Compared with control group, the lung injury in ALI group was more obvious including cellular edema and structural disorder of intercellular connection by optical microscope, and LIS, W/D ratio, and LPI were significantly increased (LIS: 3.81±0.42 vs. 0.40±0.08, W/D: 7.68±0.64 vs. 4.44±0.39, LPI: 0.89±0.15 vs. 0.38±0.05, all P < 0.01). Claudin-2 and ZO-1 were mainly expressed in the bronchial epithelium cell, and the expression degrees were significantly weakened in ALI group as compared with control group. It was shown by Western Blot results that compared with control group, the protein expressions of claudin-2 and ZO-1 were significantly down-regulated in ALI group [claudin-2 protein (gray value): 0.43±0.31 vs. 2.16±1.33, ZO-1 protein (gray value): 1.25±0.41 vs. 2.82±0.76, both P < 0.01]. Compared with ALI group, colquhounia root pretreatment could effectively diminish the degree of ALI (LIS: 1.22±0.39 vs. 3.81±0.42, W/D: 4.62±0.84 vs. 7.68±0.64, LPI: 0.46±0.07 vs. 0.89±0.15, all P < 0.01), and the protein expressions of claudin-2 and ZO-1 were significantly up-regulated [claudin-2 protein (gray value): 2.98±0.91 vs. 0.43±0.31, ZO-1 protein (gray value): 2.35±0.51 vs. 1.25±0.41, both P < 0.01]. Conclusion Administration of colquhounia root table could attenuate lung injury induced by oleic acid with improving epithelial barrier function via up-regulate the expression claudin-2 and ZO-1, which play a protective effect on the lung of rats with ALI.
3.Biosynthesis and assay of crude T-2 toxin
Yan-hong, CAO ; Shao-ping, WANG ; Yang, HUI ; Xiao-yan, FU ; Wei, DONG ; Ning, LIU
Chinese Journal of Endemiology 2008;27(5):552-554
Objective To isolate Fusarium species from Kashin-Beck disease(KBD)area and biosynthesize cnlde T-2 toxin.Methods T-2 toxin.producing Fusarium was isolated from corns produced in KBD area and purifted.The purifted funsi were identified according to the traits of colony,appearance of thallus and characters of conidium and then weIe cultivated in sterile Corn culture media.After extraction with organic solvent and purification by silica gel chromatography column,the quality and quantity of the toxin in the extracts were estimated by thin,Layer chromatography and high-performance liquid chromatography.Results The toxin-producing strain was Fusarium tricinctum. The com cuIture media inoculated with this strain produced about 250 mg of crude T-2 toxin per kg. Conclusions This experiment has indirectly further confirmed pollution of T-2 toxin-producing Fmarium existed in
4.Clinical and pathogenic analysis of Malassezia folliculitis in Nantong and Nanjing area
Yanhua PANG ; Shuanglin CAO ; Weida LIU ; Yongnian SHEN ; Mei FU ; Linling FU ; Guixia Lü ; Ping SHAO ; Wei CHEN ; Xuejun WANG
Chinese Journal of Dermatology 2012;45(2):95-98
ObjectiveTo investigate the predisposing factors and pathogenic fungal species of Malassezia folliculitis in different geographical areas and body sites.MethodsTotally,241 patients diagnosed with Malassezia folliculitis were asked to complete a questionnaire.The content of hair follicles was obtained and subjected to fungal smear and culture examination.Fungal species were identified according to morphological,physiological and biochemical features.Results Of the 241 patients with Malassezia folliculitis,204 (84.65%) were positive for smear examination.A total of 259 specimens were collected from these patients,and fungal culture grew 213(82.24%) strains,of which,209 belonged to Malassezia species,4(1.54%) to Candida species.Among the 209 Malassezia strains,186 were activated and subjected to species identification which resulted in 6 species,including M.furfur (111 strains,59.68% ),M.sloofiae (43 strains,23.12% ),M.sympodialis (17 strains,9.14%),M.globosa (9 strains,4.84%),M.pachydermatis (4 strains,2.15%),and M.obtuse(2 strains,1.08% ).Of the pathogenic fungi of Malassezia folliculitis,M.furfur predominated in the chest,back,abdomen,face and neck,M.sloofiae in the upper limbs,shoulders and vertex,M.globosa in the lower limbs.There were obvious differences in the distribution of pathogenic fungal species at different body sites in a same host,and M.furfur with M.sloofiae or M.sympodialis appeared to be the most common pathogens.ConclusionsIn this study,6 Malassezia species are identified in patients with Malassezia folliculitis in Nantong and Nanjing area,M.furfur and M.sloofiae appear to be the dominant pathogens.
5.Ester prodrug of scutellarin: synthesis, physicochemical property and degradation.
Feng CAO ; Jian-xin GUO ; Qi-neng PING ; Yun SHAO ; Jing LIANG
Acta Pharmaceutica Sinica 2006;41(7):595-602
AIMTo establish the fundamentals for the design of scutellarin prodrug and formulation with feasible physicochemical and biopharmaceutical properties by esterifying scutellarin, an active component with poor absorption extracted from Erigeron breviscapus of Chinese medicine.
METHODSWith the method of salifying followed by esterifying, ethyl and benzyl ester of scutellarin were synthesized. Glycolamide ester of scutellarin was also synthesized with an improved method. Their structures were confirmed by MS and 1H NMR. The solubility and partition coefficient of the prodrugs were determined and their degradations were investigated in various buffers and in human plasma. The emulsion and cyclodextrin complex of glycolamide ester were prepared and the protection of the ester from degradation was compared in the intestinal tract contents. Furthermore, the degradation of glycolamide ester in the homogenates of various intestinal segments was studied. Results Three prodrugs were synthesized successfully and their structures were confirmed. Glycolamide ester of scutellarin showed better stability in the aqueous solution (t(1/2) approximately =16 d, pH 4.2) and the shortest half-life in the human serum (t(1/2) approximately =7 min). Compared with scutellarin, the solubility of glycolamide ester was increased about ten times in pH 4.0 buffer, and about thirty five times in water. Partition coefficient of the glycolamide ester increased significantly from -2.56 to 1.48. However, the ester degradation in the homogenates of intestinal mucus would be an obstacle for its absorption. The degradation rates were in the order duodenum > ileum > or = jejunum > colon. The emulsion showed a better protection of glycolamide ester from the degradation than cyclodextrin complex.
CONCLUSIONGlycolamide ester of scutellarin shows better physicochemical properties than ethyl and benzyl eater of scutellarin, but its stability in intestinal tract needs to be improved. The emulsion or / and colon-targeted delivery may be selected as one of strategies to decrease the presystemic degradation.
Animals ; Apigenin ; chemistry ; isolation & purification ; pharmacokinetics ; Emulsions ; Erigeron ; chemistry ; Esters ; Flavones ; chemical synthesis ; chemistry ; pharmacokinetics ; Glucuronates ; chemistry ; isolation & purification ; pharmacokinetics ; Glucuronides ; chemical synthesis ; chemistry ; pharmacokinetics ; Humans ; Intestinal Mucosa ; metabolism ; Intestines ; metabolism ; Male ; Plants, Medicinal ; chemistry ; Prodrugs ; chemical synthesis ; chemistry ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley
6.Study on effects of Tripterygium wilfordii polycoride in resisting macrophage inflammation and regulating inflammation via TLR4/NF-κB.
Dan-ping QIN ; Yi-jun ZHOU ; Shao-zhu ZHANG ; Jun-min CAO ; Li-yu XU ; Guo-dong FANG ; Jia WANG
China Journal of Chinese Materia Medica 2015;40(16):3256-3261
To investigate the effect of Tripterygium wilfordii polycoride (TWP) on LPS-induced macrophage inflammatory response, particularly the inhibitory effect on inflammatory factors TNF-α and IL-1β and the regulatory effect on inflammation via TLR4/NF-κB. The MTT method was adopted to test the effects of tested drugs, TWP, dexamethasone (DXM) and azathioprine (AZA) on cell growth to define the appropriate concentration. LPS was used to induce the inflammatory reaction in mouse RAW264. 7 cell lines. The Elisa kit was adopted to test the release level of TNF-α and IL-1β. The Western blotting was applied to test the protein expressions of TNF-α and IL-1β. The RT-PCR was adopted to test the expressions of TLR4 and NF-κB. According to the results, TWP could inhibit the release of macrophage inflammatory factors TNF-α and IL-1β in a dose dependent manner. All of TWP groups showed a weaker efficacy than that of the DXM group. But the TWP high dose group revealed a better effect on TNF-α and equal effect on IL-1β compared with the AZA group. TWP show an equal or better effect in down-regulating TLR4 and NF-κB p65 expressions in a dose dependent manner than DXM and AZA. In conclusion, TWP could inhibit TLR4 and NF-κB p65, which may be related to the down-regulation of TLR4 and NF-κB p65 receptor expressions.
Animals
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Anti-Inflammatory Agents
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pharmacology
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Cell Proliferation
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drug effects
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Down-Regulation
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drug effects
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Drugs, Chinese Herbal
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pharmacology
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Humans
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Inflammation
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drug therapy
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genetics
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immunology
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physiopathology
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Interleukin-1beta
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genetics
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immunology
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Macrophages
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drug effects
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immunology
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Mice
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NF-kappa B
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genetics
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immunology
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RAW 264.7 Cells
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Toll-Like Receptor 4
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genetics
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immunology
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Transcription Factor RelA
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genetics
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immunology
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Tripterygium
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chemistry
7.Cloning of human vascular endothelial growth factor cDNA and its expression in COS-7 cells
Yi-Xiong ZHENG ; Li CHEN ; Shao-Ming XU ; Jia-Ping PENG ; Jiang CAO
Journal of Zhejiang University. Medical sciences 2002;31(5):316-320
OBJECTIVE: To clone vascular endothelial growth factor (VEGF) cDNA gene, construct its eukaryotic expression vector and to express this recombinant plasmid in COS-7 cells. METHODS: Human VEGF165 cDNA was amplified by RT PCR from human ovarian carcinoma. After DNA sequenced, the VEGF165 cDNA was inserted into eukaryotic expression vector pcDNA3.1(-). The recombinant plasmid pcDNA3.1 VEGF165 containing VEGF165 cDNA was identified by enzyme digestion and transferred into COS-7 cells mediated by liposome. The transient expression of VEGF was detected by immunohistochemical staining. RESULTS: The cloned VEGF165 cDNA was confirmed by enzyme digestion and DNA sequence analysis. The immunohistochemical results showed that the VEGF165 protein was expression in COS-7 cells 72 h after gene transfer. CONCLUSION: VEGF165 cDNA gene successfully cloned and expressed in COS-7 cells.
8.Study of therapeutic adjuvant-free protein vaccine-HPV16Z-Hsp65-E6/E7
Xiaobing WANG ; Mo LI ; Zhaoyang LIU ; Haimei TIAN ; Ping QU ; Yanfen LI ; Yi LIU ; Dongyan CAO ; Zhi LIANG ; Dongwan CHENG ; Changjun SHAO ; Wei ZHANG
Chinese Journal of Cancer Biotherapy 1994;0(01):-
Objective: To study the biological effects of the HPV16Z-Hsp65-E6/E7 fusion protein vaccine on the tumor associated with HPV16 infection. Methods: We tested the cellular immune responsive intensity to the vaccine by the lymphocyte proliferation and CTL response, and studied the therapeutic effect of the vaccine on mouse TC-1 cell transplanted cancer in vivo and the influence on mouse lifetime. Results: The spleen lymphocytes from the C57BL/6 mouse immunized by the Hsp65-E6/E7 vaccine could proliferate obviously in the presence of the protein and TC-1 tumor cell could be lysed specifically by immune activated lymphocytes in vitro. This animal therapeutic experiment in vivo showed that the vaccine suppressed the growth of TC-1 cell transplanted tumor remarkably. Conclusion: The recombined vaccine can induce specific cellular immune response in vitro and suppress HPV16 positive TC-1 tumor cell growth obviously in vivo.
9.Innovative ultrasound transducers-purewave crystal technology and its applications.
Shao-ping CAO ; Hong-qing GU ; Guo-jun SUN ; Jia-yi FENG
Chinese Journal of Medical Instrumentation 2005;29(5):360-367
This article is to introduce the dramatic improvements in efficiency, sensitivity and bandwidth of the ultrasound transducers--PureWave Crystal Technology.
Signal Processing, Computer-Assisted
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Transducers
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Ultrasonics
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instrumentation
10.Target volume calculation methods based on PET-CT images.
Shao-ping CAO ; Hong-qing GU ; Guo-jun SUN ; Jia-yi FENG
Chinese Journal of Medical Instrumentation 2005;29(4):273-276
The applications of PET-CT have developed from qualitative analysis to quantitative analysis. Target volume is important for tumor biological volume defining, tumor isotope therapy, organ function evaluation, acceptor affinity calculation, and pharmaceutical metabolic kinetics. Many factors work on the target volume calculation, such as PET image acquisition mode, scatter correction, attenuation correction, reconstruction method, image display mode, positron pharmacy. The commonly-used methods of target volume calculation are background-threshold, max threshold, and background-max threshold. In this article we will discuss about the methods of target volume calculation.
Algorithms
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Positron-Emission Tomography
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methods
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Radiotherapy Planning, Computer-Assisted
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methods
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Tomography, X-Ray Computed
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methods