1.Effects of moxibustion on serum IL-6, IL-8 and TNF-α in rats with experimental rheumatoid arthritis
Chuan-Ying ZHANG ; Ling HU ; Zi-Jian WU ; Lu HE ; Shuang YU
Journal of Acupuncture and Tuina Science 2019;17(5):305-311
Objective:To observe the effects of moxibustion on serum levels of interleukin (IL)-6, IL-8 and tumor necrosis factor-α (TNF-α), and to explore the effects of moxibustion on inflammatory damaging factors in experimental rheumatoid arthritis (RA) model rats; the relationship between the therapeutic effect of moxibustion on RA and the change in the Toll-like receptor (TLR) signaling pathway was analyzed using Toll-like receptor 4 (TLR4) antagonists and agonists. Methods:Fifty Sprague-Dawley (SD) rats were divided into a normal group, a model group, a moxibustion group, a moxibustion plus TLR4 agonist group (agonist group) and a moxibustion plus TLR4 antagonist group (antagonist group) according to the random number table, with 10 rats in each group. Except the normal group, rats in the other four groups were subjected to model preparation with the wind, cold and wet environmental factors plus Freund's complete adjuvant (FCA). Rats in the normal and model groups were not treated; rats in the moxibustion, agonist and antagonist groups started to be treated with the moxibustion (cigarette-type moxa) at bilateral Shenshu (BL 23) and Zusanli (ST 36) from the 4th day after the successful modeling, for 20 min each time with a total of 10 d. Rats in the agonist and the antagonist groups were injected with TLR4 agonist or antagonist [0.1 mg/(kg·bw)] via the tail vein 30 min before moxibustion. The concentrations of serum IL-6, IL-8 and TNF-α in each group were determined by enzyme-linked immunosorbent assay (ELISA). Results:Compared with the normal group, in the model group, the rat's right hind paw swelling was significantly obvious (P<0.01), there was a lot of inflammatory infiltration in the synovial tissues, the surface of the synovial membrane was unsmooth, the synovial membrane was hyperplasia and thicker, and the serum IL-6, IL-8 and TNF-α concentrations increased significantly (P<0.05). Compared with the model group, the paw swelling degrees of the rats in the moxibustion, the agonist and the antagonist groups reduced significantly (allP<0.01); the swelling degree in the antagonist group was milder than that in the agonist group, but the between-group difference was not statistically significant (P>0.05); inflammatory infiltration and synovial membrane hyperplasia in the synovial tissues of the moxibustion group and the antagonist group were all relieved differently; the decrease of synovial layer number in the moxibustion group was more obvious, and there were no obvious improvements in inflammatory infiltration and synovial thickness in the agonist group; the concentrations of IL-6, IL-8 and TNF-α in the moxibustion group were decreased, and the differences in the IL-6 and TNF-α concentrations were statistically significant (allP<0.01); there was no significant between-group difference in the IL-8 concentration (P>0.05); the concentrations of serum IL-8 and TNF-α in the agonist group increased significantly (both P<0.01), while the IL-6 concentration decreased without significant difference (P>0.05); the concentrations of IL-6 and IL-8 in the antagonist group decreased but the between-group differences were statistically insignificant (bothP>0.05), and the TNF-α concentration significantly increased (P<0.05). Compared with the moxibustion group, IL-6, IL-8 and TNF-α concentrations increased in the agonist group, and the differences in the IL-8 and TNF-α concentrations were statistically significant (both P<0.01); the concentrations of IL-6, IL-8 and TNF-α increased in the antagonist group, and the differences in the IL-6 and TNF-α concentrations were statistically significant (bothP<0.01); there was no significant difference in the IL-8 concentration between the groups (P>0.05). The serum levels of IL-6, IL-8 and TNF-α in the antagonist group were lower than those in the agonist group (allP<0.05). Conclusion:Moxibustion at Shenshu (BL 23) and Zusanli (ST 36) can reduce the joint swelling degree and inflammation in synovial tissue of RA model rats, decrease the serum levels of IL-6, IL-8 and TNF-α in RA model rats; the decreases of IL-6 and TNF-α are more significant than the decrease of IL-8; TLR4 agonist and antagonist can significantly attenuate the effect of moxibustion in inhibiting releases of IL-6, IL-8 and TNF-α, so that the change in TLR signaling pathway affects the effect of moxibustion in inhibiting the releases of IL-6, IL-8 and TNF-α.
2.Sulfur dioxide-induced liver pathology.
Chinese Journal of Pathology 2004;33(2):155-157
OBJECTIVETo investigate effects of short-term sulfur dioxide inhalation to the liver.
METHODSHaematoxylin and eosin staining (HE) and transmission electron microscopy (TEM) were used to study the pathologic changes in mice liver after sulfur dioxide (SO(2)) inhalation.
RESULTSExposure to 56 mg/m(3), 112 mg/m(3) 168 mg/m(3) SO(2) caused increasingly severe liver injuries, as detected by HE staining and TEM. The morphologic changes included spotty necrosis with lymphocyte, monocyte, and neutrophil infiltration, fatty degeneration of hepatocytes with dilatation of rough endoplasmic reticulum and dissociation of ribosomes, as well as degeneration of mitochondria and karyorrhexis.
CONCLUSIONSO(2) inhalation can cause marked liver injury in experimental settings.
Administration, Inhalation ; Animals ; Hepatocytes ; drug effects ; pathology ; Liver ; drug effects ; pathology ; ultrastructure ; Male ; Mice ; Microscopy, Electron ; Sulfur Dioxide ; administration & dosage ; toxicity
3.Chemical constituents of Taxus chinensis var. mairei cell cultures.
Xiang-yang BAI ; Jian-ming LÜ ; Yan-ying ZHOU ; Zi-rong ZHU ; Ren-wang JIANG ; Wei ZHANG
Acta Pharmaceutica Sinica 2015;50(1):70-74
The chemical constituents of Taxus chinensis var. mairei cell cultures were investigated by chromatographic methods, including silica gel column chromatography, Sephadex LH-20 and preparative HPLC. Thirteen compounds were isolated from the 80% ethanol extract of cultured cells and their structures were elucidated by spectral data and physicochemical properties, which were identified as 2α,4α,7β,9α,10β-pentaacetoxy-14β-hydroxytax-11-ene (1), 2α,4α,7β,9α,10β-pentaacetoxytax-11-ene (2), 1β-deoxybaccatin VI (3), 2α-acetoxytaxusin (4), taxuyunnanine C (5), yunnanxane (6), 2α,5α,10β-triacetoxy-14β-propionyloxy-4 (20), 11-taxadiene (7), 2α,5α,10β-triacetoxy-14β-isobutyryloxy-4 (20), 11-taxadiene (8), 2α,5α,10β-triacetoxy-14β-(2'-methyl)butyryloxy-4 (20), 11-taxadiene (9), 13-dehydroxylbaccatin III (10), 13-dehydroxy-10-deacetylbaccatin III (11), paclitaxel (12) and (13) β-sitosterol. Among them, compound 1 is a new compound, and compounds 2, 4, 10 and 11 are isolated from the cell culture of Taxus chinensis var. mairei for the first time.
Alkenes
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analysis
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Cell Culture Techniques
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Cells, Cultured
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Diterpenes
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analysis
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Molecular Structure
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Paclitaxel
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analysis
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Sitosterols
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analysis
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Taxoids
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analysis
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Taxus
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chemistry
4.Expression of DNA-PK in hepato- and cholangio-neoplasms and its significance.
Zi-jian YU ; Jian-guo SUI ; Ying-qin DING ; Zhen-shan CAO ; Ping-kun ZHOU ; De-chang WU
Chinese Journal of Hepatology 2004;12(11):652-655
OBJECTIVETo characterize DNA-PKcs and Ku70 expressions in hepato- and cholangio-neoplastic tissues and the association with the degree of malignancy and invasiveness of the tumors.
METHODSThe expression of DNA-PKcs and Ku70 was examined in 47 cases of hepato- or cholangio-neoplasm by immunohistochemistry.
RESULTSKu70 was expressed in all of the neoplastic tissues examined and with a little variation in levels. The highest expression was observed in adenocarcinomas and adenomas. There was no statistically significant association between Ku70 expression level and the degree of their malignancy extent or invasiveness. In contrast to Ku 70, a wide variation in expression levels of DNA-Pkcs was observed among different types of neoplastic tissues. The highest ratio of positive expressing cells was detected in hepatocellular carcinomas (92.1%), which was significantly higher than that in cholangioadeno carcinomas (65.3%) and biliary cystadenocarcinomas (51.9%). Low or no expression level was detected in papillary adenoma cases. DNA-PKcs expression of invasive adenomas and adeno-carcinomas (61.2%) was significantly higher than that of non-invasive adenomas and adeno-carcinomas (30.4%). There was no expression observed in the normal tissues adjacent to the tumors.
CONCLUSIONDNA-PKcs is expressed in hepato- and cholangio-neoplasms and its variable level of expression is associated with the types of the tumor and their degree of malignancy and invasiveness. DNA-PKcs could be recognized as a new biomarker for liver neoplasm.
Adenocarcinoma ; enzymology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, Nuclear ; biosynthesis ; genetics ; Bile Duct Neoplasms ; enzymology ; Bile Ducts, Intrahepatic ; enzymology ; Biomarkers, Tumor ; biosynthesis ; genetics ; Carcinoma, Hepatocellular ; enzymology ; DNA-Activated Protein Kinase ; biosynthesis ; genetics ; DNA-Binding Proteins ; biosynthesis ; genetics ; Female ; Humans ; Ku Autoantigen ; Liver Neoplasms ; enzymology ; Male ; Middle Aged
5.Application of real-time quantitative PCR in selection of transfected cell strains for transgenic overexpression.
Shao-Yan HU ; Zi-Xing CHEN ; Ye ZHAO ; Wei-Ying GU ; Jian-Nong CEN ; Jun QIAN
Journal of Experimental Hematology 2005;13(6):1062-1066
To explore the feasibility of real-time quantitative PCR (QRT-PCR) for selecting cell strains which overexpress a certain transgene, expression level of RbAp46 was detected in transfected cell strains by using optimal real-time PCR with SYBR Green I. Meanwhile, semi-quantitative RT-PCR and Western blot were performed to compare with the QRT-PCR. The results showed that values of RbAp46(N) were 2064.42 +/- 253.47, 860.94 +/- 291.07, 234.456 +/- 31.08, 18.17 +/- 5.14 and 1.46 +/- 0.54 in K562/RbAp46, K562/CMV, SHG44/RbAp46 monoclone, SHG44/RbAp46 multiclone and SHG44/CMV, respectively. The results were consistent with that determined by semi-quantitative RT-PCR and Western blot. It is concluded that QRT-PCR provides a highly efficient and reproducible method for selection of transfected cell subclones at different level of transgene expression.
Blotting, Western
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Carrier Proteins
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genetics
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metabolism
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Gene Expression Regulation, Neoplastic
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Humans
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K562 Cells
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Nuclear Proteins
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genetics
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metabolism
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Organic Chemicals
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chemistry
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RNA, Neoplasm
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metabolism
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Retinoblastoma-Binding Protein 7
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Reverse Transcriptase Polymerase Chain Reaction
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methods
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Transfection
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Transgenes
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genetics
6.Studies on chemical constituents from roots of Cudrania cochinchinensis.
Ying-Hong WANG ; Zi-Ming FENG ; Jian-Shang JIANG ; Pei-Cheng ZHANG
China Journal of Chinese Materia Medica 2007;32(5):406-409
OBJECTIVETo study the chemical constituents from the roots of Cudrania cochinchinensis.
METHODThe chemical constituents of C. cochinchinensis were isolated and purified by silica gel, polyamide column chromatography, Their structures were identified on the basis of spectroscopic data.
RESULTSix compounds were isolated and identified. Their structures were established as 3, 5, 7, 4"-tetrahydroxyflavanone-7-O-(6"-acetyl)-glucoside (1), 3, 5, 7, 4'-tetra hydroxyflavanone-7-O-glucoside (2), 2', 4', 5, 7-tetrahydroxy-6-prenyldihydroflavanone (3), 5, 7, 4'-trihysdroxy-6-prenylisoflavanone (4), 1, 3, 5, 6-tetrahydroxyxanthone (5), stilbene-2, 4, 3', 5'-tetraol (6).
CONCLUSIONCompound 1 was isolated from this genus, while compounds 4 and 5 were isolated from this plant for the first time.
Flavanones ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Moraceae ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Xanthones ; chemistry ; isolation & purification
7.Study on DNA methylation status of WT1 gene in its promoter region in hematologic malignancy cell lines.
Ye ZHAO ; Zi-xing CHEN ; Shao-yan HU ; Jian-nong CEN ; Wei-ying GU
Chinese Journal of Hematology 2005;26(9):517-520
OBJECTIVETo study the DNA methylation status of WT1 gene promoter region in hematologic malignancy cell lines and its correlation with WT1 gene expression.
METHODS1. RT-PCR and methylation-specific PCR were performed for detecting WT1 gene expression and DNA methylation status in its promoter region in 8226, HL-60, Jurkat, K562, KG-1, NB4, Raji, SHI-1, U266 and U937 cell lines. 2. Treatment of U937 cells with 5-aza-CdR, a demethylation inducing agent and the changes in WT1 gene expression level and its promoter region methylation status were determined.
RESULTS1. HL-60, K562, KG-1, NB4 and SHI-1 cells showed higher levels while 8226, Jurkat, Raji, U266 and U937 cells showed extremely low levels of WT1 expression. DNA hypermethylation in WT1 gene promoter region was identified in 8226, Jurkat, Raji, U266 and U937 cells. 2. The WT1 gene expression in U937 was enhanced after treatment with 5-aza-CdR accompanied with the decrease of methylated and the increase of unmethylated levels in its promoter region.
CONCLUSIONModulation of the DNA methylation status in WT1 promoter region is one of the epigenetic mechanisms for regulating its expression.
Cell Line, Tumor ; DNA Methylation ; Gene Expression Regulation, Neoplastic ; Hematologic Neoplasms ; genetics ; Humans ; Promoter Regions, Genetic ; genetics ; WT1 Proteins ; genetics
8.Detection of RbAp46 expression in bone marrow cells of leukemia patients by real-time quantitative RT-PCR.
Shao-yan HU ; Zi-xing CHEN ; Wei-ying GU ; Jian-nong CEN ; Ye ZHAO ; Min GU
Chinese Journal of Hematology 2005;26(7):417-420
OBJECTIVETo investigate retinoblastoma (Rb) associated protein 46 (RbAp46) gene expression levels in bone marrow (BM) cells of leukemia patients.
METHODSReal-time quantitative reverse polymerase chain reaction (QRT-PCR) method was used for detecting RbAp46 expression levels in BM cells of 140 patients with acute leukemia (AL), 13 with chronic myelogenous leukemia in chronic phase (CML-CP), 7 with CML in blast crisis (CML-BC) and 32 with non-leukemic disorders.
RESULTSThe M-Estimators of RbAp46 were higher in 98 newly diagnosed ALs and 5 relapsed ALs than in 28 ALs in complete remission (CR) and 32 non-leukemic controls (178.23 and 213.65 vs 85.89 and 88.08, respectively). No statistic difference was found between the CR group and control group, or between the newly diagnosed group and relapsed group. The M-Estimators of RbAp46 in patients with CML-CP was 58.27, similar to that in control, but much lower than that in CML-BC (173.24). Among 98 newly diagnosed ALs, the M-Estimators of RbAp46 in M(3) and M(4) were the lowest in all of the subtypes. Furthermore, the RbAp46 expression levels were not correlated with the expression of the fusion genes of bcr/abl, PML-RARalpha, and multidrug resistant gene (mdr1), but were positively correlated with Wilms' tumor gene (WT1) expression levels and negatively with AML1/ETO fusion gene expression.
CONCLUSIONRbAp46 expression levels in ALs and CML-BC were strikingly higher than that in non-leukemias and CML-CP, and might participate in leukemogenesis.
Adolescent ; Adult ; Aged ; Carrier Proteins ; genetics ; metabolism ; Child ; Female ; Gene Expression ; Humans ; Leukemia ; genetics ; metabolism ; Male ; Middle Aged ; Nuclear Proteins ; genetics ; metabolism ; Retinoblastoma-Binding Protein 7 ; Reverse Transcriptase Polymerase Chain Reaction ; methods
9.Low intensity anticoagulation therapy after mechanical heart valve replacement.
Li DONG ; Ying-kang SHI ; Zi-pu TIAN ; Jian-yang MA ; Xi WANG ; Jun YI
Chinese Journal of Surgery 2003;41(4):250-252
OBJECTIVETo investigate the proper anticoagulation intensity in patients after mechanical heart valve replacement in china.
METHODSThe anticoagulation therapy intensity and the complications in 480 patients after mechanical heart valve replacement were studied.
RESULTSThe follow-up rate was 89.17%, the total patient-years (Pty) was 2,110.04 years, the mean oral import warfarin dosage was (2.81 +/- 0.95) mg/day, and native warfarin dosage (2.38 +/- 0.46) mg/day. The mean PTR value of 2 116 samples was 1.43 +/- 0.26, and the INR value of 1 195 samples was 1.63 +/- 0.49. The total hemorrhage rate was 4.60% Pty, and the hemorrhage death rate was 0.38% Pty. The PTR and INR values were higher in the hemorrhage group than in the no-hemorrhage group. (t = 1.816, P < 0.05; t = 2.407, P < 0.01). The thromboembolism rate was 0.66% Pty, and the thromboembolism death rate was 0.05% Pty. There were 15 pregnancies in 14 women patients and no malformed newborns were found.
CONCLUSIONSThe most important complication of anticoagulation therapy after mechanical heart valve replacement is hemorrhage in china; The proper anticoagulation intensities of this group are INR 1.5-2.0 and PTR 1.3-1.5. It is beneficial to adopt the low intensity anticoagulation therapy for decreasing the death rate from hemorrhage, protecting pregnant women and newborns from hemorrhage and malformation, and improving the life qualities of the patients.
Adolescent ; Adult ; Aged ; Anticoagulants ; administration & dosage ; adverse effects ; therapeutic use ; Female ; Follow-Up Studies ; Heart Valve Prosthesis Implantation ; adverse effects ; methods ; Hemorrhage ; chemically induced ; prevention & control ; Humans ; Male ; Middle Aged ; Postoperative Care ; Postoperative Complications ; etiology ; prevention & control ; Pregnancy ; Thromboembolism ; chemically induced ; prevention & control ; Warfarin ; administration & dosage ; adverse effects ; therapeutic use ; Young Adult