2.Study on material basis of Mahuang Fuzi Xixin decoction for anti-inflammation and immune suppression based on combined method of serum pharmacochemistry and serum pharmacology.
Feng TANG ; Shao-yu LIANG ; Fei-long CHEN ; Qing-fa TANG ; Xiao-mei TAN
China Journal of Chinese Materia Medica 2015;40(10):1971-1976
To investigate me material basis of Mahuang Fuzi Xixin decoction (MFXD) for anti-inflammation and immune-suppression based on the combined method of serum chemical and serum pharmacological. The LC-MS/MS fingerprints of MFXD, drug-containing serum and blank serum were compared to define the components in plasma. Histamine, β-hexosaminidase released from RBL-2H3 cell infulenced by drug-containing serum at different time points were measured by ELISA. The effect of drug-containing serum on lipopolysaccharide-induced splenocyte proliferation at different time points were determined by MTT. A correlation analysis was made on components of MFXD and pharmacological indexes based the stepwise regression method. After the intragastrical administration with MFXD, 32 components were discovered in rat serum, including 27 prototype components (10 from Mahuang, 13 from Fuzi and four from Xixin) and five unknown components. Compared with blank serum, drug-containing serum could reduce the release of histamine from RBL-2H3 induced by antigen at different time points (P < 0.05); except the 4-hour drug-containing serum, all of the remaining drug-containing serums could inhibit the RBL-2H3 mastocyte degranulation induced by antigen at different time points (P < 0.05). Drug-containing serum could significantly lipopolysaccharide-induced mouse splenocyte proliferation at 15 and 30 min (P < 0.05). A regression analysis was made on the chemical data of components absorbed into blood and pharmacological indexes, i. e. release rate of histamine, release rate of β-hexosaminidase and inhibition rate of splenocyte. This suggested the close correlations among methyl pseudo-ephedrine, pseudoephedrine and histamine released from RBL-2H3 induced by antigen; pseudoephedrine, hypaconine, methyl pseudoephedrine and β-hexosaminidase released from RBL-2H3 induced by antigen; as well as benzoyl hypaconine, benzoylaconine, 14-benzoyl-10-OH-mesaconine, mesaconine and lipopolysaccharide-induced mouse splenocyte proliferation. Methylpseudoephedrine, pseudoephedrine, benzoyl hypaconine, benzoylaconine and mesaconine may be part of material basis of MFXD on anti-inflammation and immune suppression.
Animals
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Anti-Inflammatory Agents
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chemistry
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pharmacology
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Cell Degranulation
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drug effects
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Drugs, Chinese Herbal
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chemistry
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pharmacology
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Female
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Histamine
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immunology
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Immunosuppressive Agents
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chemistry
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pharmacology
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Male
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Mass Spectrometry
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Mast Cells
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drug effects
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immunology
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Mice
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Rats
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Rats, Wistar
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Serum
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chemistry
3.Relationship between combined multigene detection and response to chemotherapy and prognosis in epithelial ovarian carcinomas
Li LI ; Xiu-Ying MO ; Wei ZHANG ; Min-Yi WEI ; Fa-Long CHEN ; De-Sheng YAO ;
Chinese Journal of Obstetrics and Gynecology 2000;0(09):-
Objective To evaluate the relationship between combined muhigene detection and response to chemotherapy and prognosis in epithelial ovarian carcinomas(EOCS).Methods A total of 80 ovarian tissue samples taken from the surgical specimens of patients with EOCS of our hospital in the last two decades who had received chemotherapy "after surgery were paraffin-embedded.The samples were divided into 2 groups,good prognosis group(patients who survived more than 2 years,n=46)and poor prognosis group(patients who survived less than 2 years,n=34).The expression levels of ToPo-Ⅱ,Ki-67,MGMT, PCNA,p27,p53,pl6,P-gp,LRP,GST-?,bcl-2,C-myc,Fas,bax,MSH2,MRP and BCRP were investigated by the combination of tissue arrays and immunohistoehemical streptavidin-biotin peroxidase(SP) method in all samples.Data were analysed with SPSS 12.0 for windows.Results There were statistically significant differences in the positive expression levels of P-gp,BCRP,MGMT,MSH2,p27 and p16(62%, 50% and 50% in poor prognosis group vs 33%,28% and 28% respectively,P
4.Investigation report of the effect of control measures to iodine deficiency disorders in Xining in 2009
Li-lin, CHEN ; Duo-long, HE ; Shu-bang, LI ; Fa-rong, ZHANG ; Xun, CHEN ; Pei-chun, GAN ; Zhi-jun, ZHAO
Chinese Journal of Endemiology 2011;30(1):81-83
Objective To master iodine nutritional status of people after universal salt iodization in Xining that reached the stage goal of elimination iodine deficiency disorders. Methods In the 7 counties investigated of Xining in 2009, 5 towns were randomly selected in each county, and one school was randomly selected in each town, 80 children aged 8 to 10 were randomly selected in each school, and goiter were examined, urinary iodine and salt iodine were tested. Thyroid gland goiter of children was detected by thyroid palpation, children's urinary iodine was tested by As( Ⅲ )-Ce4+ catalytic spectrophotometry, and salt iodine was tested by direct titration. Results A total of 2919 children aged 8 to 10 were examined, 31 goiter was detected, goiter rate was 1.06%(31/2919).One thousand and seventy-eight urine samples were detected, urinary iodine median was 205.3 μg/L, that lower than 20 μg/L accounted for 1.9% (20/1078), lower than 50 μg/L accounted for 4.5%(48/1078). Two thousand and seventy-nine salt samples were detected, median of salt iodine was 32.80 mg/kg, the rate of non-iodized salt was 0.87%(18/2079), the coverage rate of iodized salt was 99.13%(2061/2079), the qualified rate of iodized salt was 98.64% (2033/2061), the consumption rate of qualified iodized salt was 97.79% (2033/2079). Conclusions Prevention and control of iodine deficiency disorders has achieved remarkable results in Xining city, all indicators have reached the national standard to eliminate iodine deficiency disorders.
5.Construction of a novel Schistosoma japonicum DNA vaccine pBK-Sj14-3-3 and studies on its immunoprotection in mice.
De-fa LI ; Yue-sheng CHEN ; Ying ZU ; Ji-long SHEN
Chinese Journal of Preventive Medicine 2004;38(3):193-195
OBJECTIVETo prepare Sj14-3-3 DNA vaccine and observe its immunoprotection against Schistosoma japonicum in mice.
METHODSThe Sj14-3-3 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and subcloned into eukaryotic expression vector pBK. The recombinant plasmid pBK-Sj14-3-3 was extracted, purified and inoculated into BALB/c mice by intramuscular injection. Mice were attacked by Schistosoma japonicum cercariae and then killed. Adult worm and egg were counted, respectively. Diameter of the egg granulomas in the liver of infected mice was measured.
RESULTSElectrophoresis on 1% agarose gel showed that the product of RT-PCR and the inserted fragment of recombinant plasmid digested with EcoR I and Xho I had the same size, about 765 bp, confirming the latter was the 14-3-3 encoding gene by nucleotide sequencing. Adult worm load declined by 27%, average egg load of per gram (EPG) of the liver tissues by 79%, average egg production per couple of adult worm (EPWP) by 51%, and mean diameter of egg granulomas by 29% in vaccinated mice.
CONCLUSIONThe recombinant plasmid pBK-Sj14-3-3 was successfully constructed, which had some immunoprotection against Schistosoma japonicum in infected mice, indicating its potential to be vaccine candidate molecule of Schistosoma japonicum.
14-3-3 Proteins ; genetics ; immunology ; Animals ; Antibodies, Helminth ; blood ; Antigens, Helminth ; genetics ; immunology ; Cloning, Molecular ; DNA, Helminth ; genetics ; Female ; Helminth Proteins ; genetics ; immunology ; Membrane Proteins ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Parasite Egg Count ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Schistosoma japonicum ; genetics ; immunology ; Schistosomiasis japonica ; immunology ; prevention & control ; Vaccines, DNA ; immunology
6.Small interfering RNA targeting Apollon enhances the chemosensitivity of hepatocellular carcinoma cells in vitro.
Jian-fa CHEN ; Yu-hua LI ; Yin-xiang CHEN ; Kui-long XIE ; Ming FU ; Li LI
Journal of Southern Medical University 2011;31(10):1701-1704
OBJECTIVETo investigate the effect of small interfering RNA (siRNA) targeting Apollon in enhancing the chemosensitivity of hepatocellular carcinoma (HCC) cells in vitro.
METHODSHCC cells transfected with the siRNA targeting Apollon were tested for Apollon protein expression using Western blotting. MTT assay and ELISA were used to evaluate the proliferation and apoptosis of HCC cells transfected with siRNA after exposure to 5-FU or adriamycin.
RESULTSApollon siRNA obviously down-regulated Apollon protein expression in HCC cells. The siRNA-mediated suppression of Apollon expression resulted in a marked inhibition of cell growth and increased apoptotic rate of HCC cells, and enhanced both the growth-inhibiting and apoptosis-inducing effects of the chemotherapeutic drugs.
CONCLUSIONApollon siRNA can enhance the chemosensitivity of HCC cells to the chemotherapeutic drugs. Apollon can be a potentially important and feasible therapeutic target for HCC.
Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Apoptosis ; genetics ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Hep G2 Cells ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Liver Neoplasms ; pathology ; RNA Interference ; RNA, Small Interfering ; genetics
7.Application of SEMG to study the effects of imagery training on back-style high jump.
Wen-Feng LIU ; Yong-Ling CHANG ; Chang-Fa TANG ; Zhen-Zhen HONG ; Li-Qin YIN ; Jin CHEN ; Wen-Ning REN ; Long JIANG ; Jian KUANG
Chinese Journal of Applied Physiology 2013;29(3):260-270
Adolescent
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Adult
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Athletic Performance
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psychology
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Back
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physiology
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Electromyography
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Exercise
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physiology
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Humans
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Imagery (Psychotherapy)
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Male
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Young Adult
8.Detection of IL-35-expressing monocytes in mice infected with BCG and its clinical significance
Chen CHEN ; Jun-Ai ZHANG ; Yu-Qing LIU ; Huan XU ; Hou-Long LUO ; Rui-Xi LI ; Bi-Ying ZHENG ; Jun-Fa XU
Chinese Journal of Immunology 2018;34(2):172-176
Objective:To detect IL-35 in mononuclear cells of peripheral blood and lung tissue with BCG infected mouse,and to analyze its effect on the tuberculosis immune mechanism and pathology.Methods: Mycobacterium tuberculosis infecting animal model was made by BCG injection in wild type C57BL/6 mouse.Then,mice were sacrificed in different time points.The peripheral blood and lung tissue were isolated.The bacterial load and histopathological analysis in lung tissue were performed,and the expression of IL-35 subunits P35 and EBI3 in monocytes were detected by flow cytometry.Furthermore,the correlation of IL-35-expressing monocytes with the load of bacterium was analyzed.Results:The expression of P35 and EBI3 in monocytes in peripheral blood and lung tissue of BCG-infected mice was significantly higher than that of the control group,and the peripheral blood of the experimental group was significantly increased at 4 weeks after infection.The expression of EBI3 in lung tissue diaplayed significant rising trend.The correlation analysis showed that the P35 expression was positively correlated with the expression of EBI3 in monocyte,and the IL-35-ex-pressing monocyte in lung tissue was negatively correlated with the load of bacteria.The IL-35-expressing monocyte in peripheral blood and lungs increased significantly at 2-week and 4-week,while it increased in lung tissue at 8 week,significantly.Conclusion: BCG-infects mouse monocytes with high expression of IL-35,it may participate in anti-tuberculosis immune regulation.
9.Development and evaluation of gamma-interferon sandwich ELISA kit in herds infected with Mycobacterium bovis
Xin LI ; Zheng-Zhong XU ; Fa SHAN ; Ai-Hong XIA ; Chuang MENG ; Ye-Chi SHEN ; Yi-Ping CHEN ; Wen-Long NAN ; Xiang CHEN ; Xin-An JIAO
Chinese Journal of Zoonoses 2017;33(12):1060-1065
This study was aimed to develop a sandwich ELISA kit for the diagnosis of bovine tuberculosis.And it was applied and evaluated in the quarantine of bovine tuberculosis.We established a bovine IFN-γ release method in vitro and developing three batches of kits.The sensitivity,repeatability and retention period of the kit were all evaluated.Totally 961 serum samples were tested using the developed sandwich ELISA kit tuberculin skin test and a commercial ELISA kit.Our results showed that the detection limit of this ELISA was 8.21 mg/mL.The repeatability tests showed good reproducibility in the intraassay and inter-assay.At the same time,the retention period of the kit was more than 12 months.Compared with the tuberculin skin test,the positive coincidence rate was 70.59% and the negative coincidence rate was 99.20%,while the total coincidence rate was 98.44%.And compared with the BOVIGAMTM kit,the positive coincidence rate was 91.30% and the negative coincidence rate was 99.78%,while the total coincidence rate reached 99.58%.At the same time,the sensitivity and specificity of the sandwich kit were 85.00% and 100%,respectively.We established a bovine IFN-γ release method in vitro and developing corresponding kits successfully have a good application prospect.
10.Effects of xuefuzhuyu injection on expression and activation of NF-кB in rat brain tissue after traumatic brain injury
Bing CHEN ; Yan-Qing YIN ; Hao PENG ; Hua-Wei WU ; Yuan-Sheng LIANG ; Xiao-Ao LONG ; Jun-Fa XU ; Dong CHEN
Chinese Journal of Neuromedicine 2010;9(1):15-19
Objective To investigate the effects of xuefuzhuyu injection on the expressions of the nuclear factor-кB(NF-кB)and inhibitor-кBα(1-кBα)in rat brain tissue after traumatic brain injury.Methods Eighty SD rats were randomly divided into sham-operative(n=20), model(n=20),hexadecadrol-treated(n=120)and xuefuzhuyu-treated(n=20)groups.Traumatic brain injury models were induced by free-fall.Sham-operative and model groups received intraperitoneal injection of 0.1 mL/(kg·d)normal saline;dexamethasone-treated group and xuefuzhuyu-treated group received intraperitoneal injection of 0.1 mg/(kg·d)dexamethasone and 4 mL/(kg·d)xuefuzhuyu injection immediately after the successful model inducement,respectively, and then, heir brain tissue samples were obtained at 6,24,48 and 96 h. Immunohistochemistry was employed to detect the protein expressions of NF-кB P65 and I-кBα in the traumatic brain tissue,and HE staining was performed to observe the brain structure.Results Increased protein expression of NF-кB P65 and decreased protein expression of I-кBα in the model group were found as compared to those in the sham-operative group at all time points(P<0.05);dexamethasone-treated and xuefuzhuyu-treated groups showed significantly lower protein expression of NF-кB and higher protein expression of I-кBα than the model group at all time points(P<0.05)and the effect of xuefuzhuyu injection was better than that of dexamethasone.The expressions of NF-кB and I-кBα were negatively correlated in the model group(r=-0.876.P=0.000). Conclusion Secondary injury caused by traumatic brain injury can be relieved by inhibiting the activation of NF-кB and increasing the expression of I-кBα, which are exactly the effects of xuefuzhuyu injection and dexamethasone,and xuefuzhuyu injection works better than dexamethasone.