1.Bibenzyl from Dendrobium inhibits angiogenesis and its underlying mechanism.
Chenyuan GONG ; Bin LU ; Li YANG ; Lei WANG ; Lili JI
Acta Pharmaceutica Sinica 2013;48(3):337-42
Bibenzyl is a type of active compounds abundant in Dendrobium. In the present study, we investigated the inhibitory effects of six bibenzyls isolated from Dendrobium species on vascular endothelial growth factor (VEGF)-induced tube formation in human umbilical vascular endothelial cells (HUVECs). All those bibenzyls inhibited VEGF-induced tube formation at 10 micromol x L(-1) except tristin, and of which moscatilin was found to have the strongest activity at the same concentration. The lowest effective concentration of moscatilin was 1 micromol x L(-1). Further results showed that moscatilin inhibited VEGF-induced capillary-like tube formation on HUVECs in a concentration-dependent manner. Western blotting results showed that moscatilin also inhibited VEGF-induced phosphorylation of VEGFR2 (Flk-1/KDR) and extracellular signal-regulated kinase 1/2 (ERK1/2). Further results showed that moscatilin inhibited VEGF-induced activation of c-Raf and MEK1/2, which are both upstream signals of ERK1/2. Taken together, results presented here demonstrated that moscatilin inhibited angiogenesis via blocking the activation of VEGFR2 (Flk-1/KDR) and c-Raf-MEK1/2-ERK1/2 signals.
2.Establishment of a rat model of blood hypercoagulable state caused by intravenous injection of thrombin
Liwen WANG ; Xiaojie SHEN ; Qian WU ; Yingying JI ; Guoqing GONG
Acta Laboratorium Animalis Scientia Sinica 2016;24(6):639-642
Objective The aim of this study was to establish a rat model of blood hypercoagulable state by intra?venous injection of thrombin and to provide a model for researches on hypercoagulable state. Methods Rats were divided into six groups and were injected with normal saline and 2?5, 5, 10, 20, 40 U/kg thrombin solution through the femoral vein, respectively. Then, blood was drawn to test the activated partial thromboplastin time (APTT), prothrombin time ( PT) and fibrinogen ( FIB) , and to observe the death rate of rats in these groups to verify the optimal dosage. On this ba?sis, rats were injected thrombin of the best dose through the femoral vein, and blood samples were collected at 0, 10, 30, 60, 120, 180, 300 (s) to test APTT and PT and FIB for determining the best time for blood sampling. At last, the rats were divided into control group and thrombin group to inject normal saline or thrombin solution in the best dose via the fem?oral vein, and blood was taken at the best time to test APTT, PT, FIB and whole blood viscosity. Results APTT and PT values of the 10 U/kg thrombin group were the shortest, and FIB value of this group was the highest among these groups. APTT and PT values of blood sample collected at about 60 s after thrombin injection were the shortest, and FIB value was the highest. Compared with the control group, PT and APTT values of the thrombin group were shorter (P<0?05), and blood viscosity and FIB were higher ( P<0?05 ) . Conclusions Injecting thrombin solution into the femoral vein can be used to establish a rat model of hypercoagulable state. The best dose of thrombin solution is 10 U/kg in a concentration of 2 U/mL. The best time to collect blood sample is 60 s.
3.The analysis of key indicators what may affect the acticity index in systemic lupus erythematosus
Wuping LI ; Haiying WANG ; Xia YANG ; Yi GONG ; Mei JI
Chinese Journal of Rheumatology 2012;16(5):343-345
Objective To investigate the distribution of characteristics,clinical manifestations,laboratory parameters and activity index of patients with systemic lupus erythematosus (SLE) and to determine their role in helping to make correct clinical diagnosis and disease the activity evaluation of SLE.Methods Collect the data of 1037 SLE patients of Ningxia Medical University Affiliated Hospital from January 2006 to June 2010.Data were analyzed with t test and Chi-square test.Results Over the past three years,there were more and more patients were admitted year by year.Among the 1037 cases of SLE patients,most of them 20-40 year-old woman,accounting for 67.5% of the whole patient population,with a male to female ratio was 1:8.26.Joint pain was the most common initial symptom,accounting for 54.3%,followed by skin rash,accounting for 48.2%.Decreased complement C3 level and platelets counts, proteinuria,and positive anti-dsDNA antibody could be used as indicators for early diagnosis of SLE.SLEDAI activity score higher than 9 were presented in 26.0% of patients.Factors that could impact the final score of SLEADI were fever,arthritis,skin rash,proteinuria,low complement levels,high titers of anti-dsDNA antibody,pleurisy,alopecia,mucosal ulcers,pericarditis,mental illness and decreased platelets count.Patients with active disease had a higher accidence of fever,arthritis,skin rash,lung damage,alopecia,mucosal ulcers,heart damage,mental illness and renal damaged,low complement levels,high level of anti-dsDNA antibody titers and elevated erythrocyte sedimentation rate.Conclusion SLE is a multi-system disease with multiple organ involvement,with characteristic clinical symptoms and immunological abnormalities,thus early diagnosis is very important.Understanding the characteristics of the diseases,correct judgement of the disease activity,reasonable and effective treatment all can delay the development of organ damage and improve the prognosis.
4.Dynamic changes of five constituents in Ligustri lucidi Fructus at five picking time
Fucheng WANG ; Donghan JI ; Chengwu FANG ; Daofeng GONG ; Chengkai XIA
Chinese Traditional Patent Medicine 2017;39(5):1011-1014
AIM To analyze the dynamic changes of five constituents in Ligustri lucidi Fructus at five picking time (August,September,October,November,December).METHODS The HPLC analysis of Ligustri lucidi Fructus ethanol extract was performed on a 25 ℃ thermostatic Aglient Zorbax SB-C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1% phosphoric acid flowing at 1 mL/min in a gradient elution manner,and the detection wavelength was set at 224 nm.RESULTS Salidroside,tyrosol,luteolin-7-O-glucoside,ligustroflavone and specnuezhenide showed good linear relationships within their own ranges (r >0.999 0),whose average recoveries were 99.56%-100.30% with the RSDs of 0.89%-1.23%.The contents of various constituents (except for tyrosol) were the highest in samples picked up in September,followed by those picked up in October.CONCLUSION The suitable picking time of Ligustri lucidi Fructus is September and October.
5.CT diagnosis on rupture of hepatic echinococcosis into the biliary tract
Rui GONG ; Ji YIN ; Feng WANG ; Najia LIU ;
Chinese Journal of Radiology 2001;0(04):-
Objective To analyze the CT features of hepatic echinococcosis rupture into the biliary tract, and to assess the value of CT study in the diagnosis of such cases. Methods CT findings in 15 cases proved surgically and pathologically were studied and analyzed retrospectively. Results 15 patients had 20 hepatic echinococcosis, including simple echinococcosis with single cyst (9 lesions), simple hydatid cyst with multiple cysts (6 lesions), and multivesicular echinococcosis with multiple cysts (5 lesions). 2 lesions had calcifications on the cyst wall. The rupture of hepatic echinococcosis into the intrahepatic ducts was found in all 15 cases. Meanwhile, 12 of them had rupture into the common bile duct, and 2 into the gallbladder. The main CT findings were hepatic echinococcosis incorporated with the dilatation of the biliary tract. Conclusion Signs of rupture of hepatic echinococcosis into the biliary tract are characteristic in CT studies, providing a reliable evidence for preoperative diagnosis.
6.Effects ofZi-Bu Pi-YinRecipe on Dendritic Spines in Spleen-YinDeficiency Dementia Model Rats
Hua SUI ; Libin ZHAN ; Zhenghong JI ; Xiaoyang GONG ; Jin GONG ; Jian WANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(10):2029-2033
This study was aimed to observe the changes of dendritic spine density in different regions of brain among spleen-yindeficiency dementia (SYDD) model rats, in order to investigate the effects ofZi-Bu Pi-Yin Recipe (ZBPYR) on dendritic spines. Spleen-yindeficiency (SYD) rats were modeled by classical method. And incubatedβ-Amyloid 1-40 (Aβ1-40) was injected into the hippocampus of each rat to make SYDD model, which received the administration of ZBPYR. Golgi staining was used to stain dendritic spine in different regions of brain in rat model for the observation of the amount and shape. The results showed that dendritic spine density in different regions of hippocampus and cortex in SYDD group was reduced than that of the SYD group. Compared with the dementia group and the SYDD group, the dendritic spine density in different regions of hippocampus and cortex of the SYDD + ZBPYR group was increased. Compared with the blank control group, the dendritic spine density in different regions of hippocampus and cortex in rats from the dementia group was reduced. It was concluded that there were different degrees of reducing in the dendritic spine density of different brain regions in SYDD group. ZBPYR improved the learning and memory impairment, which might be related to the maintenance of dendritic spine density in different brain regions.
7.Effects of recombinant soluble MICA protein on the biologic activities of NK cells
Weijuan GONG ; Haiyang WANG ; Minqi FAN ; Chunxiang GONG ; Dan LIU ; Mingchun JI
Journal of Cellular and Molecular Immunology 2009;25(10):903-906
AIM: To study the effects of recombinant soluble MHC class Ⅰ chain-related protein A (sMICA) on the cytotoxicity, secretion of IFN-γ, proliferation and apoptosis of peripheral NK cells. METHODS: After NK cells were cocultured with recombinant soluble MICA proteins overnight, the cytotoxicity of NK cell on target cells was detected by flow cytometry. The supemant was collected to determine the concentration of IFN-γ by ELISA. The proliferation of NK cells to sMICA was detected by MTS/PMS. NK cells were labeled with annexin V and PI to analyze their apoptosis. RESULTS: Soluble MICA inhibited the cytotoxicity of NK cells and down-regulated the secretion of IFN-γ, but it showed no effects on the proliferation and apoptosis of freshly isolated peripheral NK cells. CONCLUSION: The soluble MICA shedding from tumor cells could be a pathway of cancer immune evasion by down-regulating the biologic activities of NK cells.
8.Microbial monitoring analysis of methylcellulose eye drops during use and storage
Ruizhong, GONG ; Hong, ZHANG ; Li, WANG ; Junfeng, WANG ; Jing, WANG ; Lin, ZHANG ; Ming, JI
Chinese Journal of Experimental Ophthalmology 2015;33(9):816-819
Background It is imperative for the microbial monitor after opening the bottle of eyedrops in order to ensure the safety during use of ophthalmic solutions with multi-dose packaging.Objective This study was to research the microbiological properties and sterile duration of methylcellulose (MC) eye drops in three common environmental conditions,including room temperature condition of community,refrigeration condition of community and room temperature condition of hospital.Methods MC eye drops were assigned to the community room temperature group,community refrigeration group and hospital room temperature group,and 200 bottles of MC eye drops with or without ethylparaben were collected in each group,including sealed or unsealed drugs at average.The containers of all the eye drops were opened and the opening times were record.The drugs was admistered 1 drop for 3 times per day,with the opening period for 5-10 seconds.Then the drugs were preserved in different environments based on grouping.Microbial isolation and purification were performed by the same lab technician at 8:00 from 1 through 10 days after opening of drugs with automatic microbial analyzer.Results In the unsealed MC eye drops without ethylparaben,the bacterial positive rates were about 30% in the community room temperature group,community refrigeration group and hospital room temperature group,but no microbial colony was seen in the sealed eye drops.Ten days after opening of containers,the bacterial cultured rates were 30%,32% and 36% in the eye drops without ethylparaben in the community room temperature group,community refrigeration group and hospital room temperature group,and those in the eye drops with ethylparaben were 15%,19% and 23%,respectively,showing significant differences between the eye drops with and without ethylparaben (x2 =6.452,4.448,4.063,all at P<0.05).The 95% confidence interval (CI) of difference values of intergroup bacterial rates were-0.166-0.126,-0.110-0.190 and-0.088-0.208 between the community room temperature group and the community refrigeration group,between the hospital room temperature group and the community refrigeration group,between the hospital room temperature group and the community room temperature group respectively in the unsealed eye drops without ethylparaben,and those in the unsealed eye drops with ethylparaben were-0.159-0.079,-0.089-0.169 and-0.043-0.203 respectively,indicating insignificant differences among the groups.Cultured bacteria were identified as Micrococcus luteus,Acinetobacter lwoffii,Bacillus subtilis,Acinetobacter radioresistens,Myroides and Staptococcus xylosus.Conclusions Ethylparaben can reduce the contamination rate of microorganisms after opening of MC eye drops.Three environmental conditions do not play an influence on microbial contamination of MC eye drops after opening.The bacteria of contaminated eye drops appear to be common microorganisms in atmosphere and soil,rather than eye common pathogens.
9.Isolation and identification of Japanese encephalitis virus in the experimental minipigs
Ji WANG ; Rui FU ; Xiaobo LI ; Shujing WANG ; Wei GONG ; Li WEI ; Bingfei YUE ; Zhengming HE
Chinese Journal of Comparative Medicine 2017;27(3):57-62
Objective To understand the characteristics of minipigs infected withJapanese encephalitis virus(JEV).Methods After the brain tissues were treated, the pig brain tissue treatment solution was inoculated with BHK21 cells.Then, virus culture,indirect immunofluorescence assay, neutralization test, electron microscopic observation, and reverse transcription-polymerase chain reaction (RT-PCR) amplification of the new isolate E segment and PrM segment nucleotide sequence were performed and the genotype was identified.Results BHK21 cells were inoculated into 25 pigbrain tissues.Among them, three tissue-treated fluid couldinduce shrinkage and aggregation of BHK21 cells, and immunofluorescence staining showed strong green fluorescence response.The results of neutralization test showed that the neutralization titer of these three new isolates was 1:64, and the size of the virus particles was about 40nm under the electron microscope.The homology of both RT-PCR product sequencing results and E-segment of vaccine strain were 95%.Three new isolates were type GIII JEV.Conclusion The results ofthisstudydemonstrate that there is G III type Japanese encephalitis virus infection in the minipig farm.
10.Detection of chicken embryo lethal orphan virus and egg drop syndrome virus by multiplex polymerase chain reaction
Shujing WANG ; Rui FU ; Xiaobo LI ; Ji WANG ; Li WEI ; Wei GONG ; Bingfei YUE ; Zhengming HE
Chinese Journal of Comparative Medicine 2015;(1):66-70
Objective To establish multiplex PCR assay for detection of chicken embryo lethal orphan virus (CELO)and egg drop syndrome virus (EDS).Methods According to GenBank gene sequence , two pairs of specific primers designed were amplified CELO long fiber protein and EDS hexon protein gene sequence .The specificity and sensitivity of multiplex PCR were tested .We also use the multiplex PCR to detect exogenous CELO and EDS in influenza virus.Results Two target bands have been successfully amplified and verified by sequencing .The specificity of the method is better , and the sensitivity is 10 -4μg/mL.The results of detecting exogenous CELO and EDS in 12 influenza virus were negative .Conclusion The multiplex PCR assay for detection of CELO and EDS was established successfully , which have good specificity and high sensitivity , and have high value and application prospect for detecting exogenous CELO and EDS in influenza virus .