3.Exploration of the mechanism of xinfeng capsule in the treatment of ankylosing spondylitis based on B and T lymphocyte attenuator and oxidative stress.
Ya-Jun QI ; Jian LIU ; Li ZHENG ; Yun-Xiang CAO ; Lei WAN
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(1):25-32
OBJECTIVETo explore changes of B and T lymphocyte attenuator (BTLA), superoxide dismutase (SOD), catalase (CAT), total antioxidant capacity (TAOC), reactive oxygen species (ROS), reactive nitrogen species (RNS), malondialdehyde (MDA) in ankylosing spondylitis (AS) patients, and the effect of Xinfeng Capsule (XFC) on them.
METHODSTotally 120 AS patients were assigned to two groups according to random digit table method, the XFC group (3 XFC pills each time, 3 times a day) and the SASP group (4 SASP tablets each time, twice a day), 60 in each group. All patients were treated for 3 months. Another 60 healthy subjects were recruited as a healthy control group. The expression frequency and activation levels of BTLA were detected using flow cytometry. Serum oxidative stress indices (such as SOD and CAT, TAOC, ROS, RNS, MDA) and contents of cytokines [tumor necrosis factor α (TNF-α), IL-1β, IL-4, and IL-10] were detected using enzyme-linked immunoassay (ELISA). Erythrocyte sedimentation rate (ESR) was detected using Westergren method. High-sensitivity C-reactive protein (Hs-CRP) was detected using HITACHI 7060 type automatic biochemical analyzer. Clinical efficacies of ASAS 20 and BASDAI50 were assessed using VAS. Correlation analysis between scoring for quality of life and BTLA expression frequency was performed.
RESULTS(1) Clinical efficacies of ASAS 20 and BASDAI50 were significantly better in the XFC group than in the SASP group (P < 0.01). (2) Compared with the healthy control group, BTLA expressions in the peripheral blood of AS patients decreased significantly (P <0. 05); SOD, CAT, and TAOC values significantly decreased (P < 0.01, P < 0.05); ROS, RNS, and MDA values significantly increased (P < 0.01, P < 0.05); TNF-α, IL-1β, ESR, and Hs-CRP values significantly increased (P < 0.01); IL-4 and IL-10 values decreased significantly (P < 0.01, P < 0.05). (3) Compared with pre-treatment in the same group, BTLA/CD19 + B, BTLA/CD24 + B, SOD, TAOC, IL-4, SF-36 [physical functioning (PF), social functioning (SF), role limitation due to physical problems (RP), role limitation due to emotional problems (RE), body pain (BP), mental health (MH), vitality (VT), general health (GH)] were significantly elevated; ROS, MDA, TNF-α, ESR, Hs- CRP, VAS, BASDAI and BASFI, BAS-G were significantly lower in the peripheral blood of the two groups after treatment (P < 0.01, P < 0.05). Better effect was shown in the XFC group in elevating BTLA/CD19+ B, BTLA/CD24 + B, SOD, TAOC, IL-10, BP, MH, VT, and SF; and lowering ROS, IL-1β, MDA, TNF-α, ESR, Hs-CRP, VAS, BASDAI, BASFI, and BAS-G (P < 0.01, P < 0.05). (4) Pearson correlation analysis showed, BTLA/CD19 + B expression of the peripheral blood was positively correlated with SOD, CAT, TAOC, IL-4, IL-10, GH, RP, BP, and SF (r = 0.431, 0.325, 0.318, 0.316, 0.348, 0.314, 0.358, 0.318, 0.326, respectively, P < 0.05, P < 0.01), while it was negative correlated with ROS, MDA, TNF-α, IL-1β, ESR, VAS, and BASDAI (r = -0.342, -0.368, -0.334, -0.354, -0.324, -0.372, -0.342, respectively, P < 0.05, P < 0.01). BTLA/CD24 B expression of the peripheral blood was positively correlated with SOD, TAOC, IL-4, IL-10, GH, RP, BP, SF, RE, MH, VT (r = 0.358, 0.352, 0.372, 0.436, 0.435, 0.326, 0.352, 0.345, 0.326, 0.343, 0.332, respectively, P < 0.05, P < 0.01), while it was negative correlated with ROS, RNS, MDA, ESR, Hs-CRP, VAS, BASDAI, and BASFI (r = -0.447, -0.336, -0.405, -0. 395, -0. 358, -0.436, -0.338, -0.425, respectively, P < 0.05, P < 0.01).
CONCLUSIONXFC could improve BTLA expression in the peripheral blood of AS patients, negatively regulate activation and proliferation of B cells, and reduce abnormal immune responses and oxidative stress injury, thereby effectively alleviating joint stiffness and pain.
B-Lymphocytes ; physiology ; Blood Sedimentation ; C-Reactive Protein ; metabolism ; Capsules ; Catalase ; metabolism ; Cytokines ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Flow Cytometry ; Humans ; Interleukin-10 ; metabolism ; Interleukin-1beta ; metabolism ; Interleukin-4 ; metabolism ; Malondialdehyde ; metabolism ; Oxidative Stress ; Quality of Life ; Reactive Oxygen Species ; Spondylitis, Ankylosing ; drug therapy ; Superoxide Dismutase ; metabolism ; T-Lymphocytes ; physiology ; Tumor Necrosis Factor-alpha ; metabolism
4.Establishment and evaluation of methods for determinating cystic fibrosis transmembrane conductance regulator quantitatively.
Feng QIU ; Jie ZENG ; Kun LI ; Ai-jun CHEN ; Wan-xiang XU ; Ya NI
Chinese Journal of Applied Physiology 2015;31(2):154-157
OBJECTIVETo establish and evaluate a BA-ELISA method for the quantitative detection of cystic fibrosis transmembrane conductance regulator (CFTR) protein.
METHODSWe deliberately selected three tables of CFTR and made the synthetic peptide be expressed in E. coli, then used the antigen to immunize rabbits to obtain the anti-CFTR polyclonal serum. After that, 96 well plates were coated with the purified antibody against CFTR. The antigen CFTR which was extracted from human sperm was detected by anti-CFTR antibody labeled with biotin, horseradish peroxidase conjugated avidin, and the substrate. The concentrations of two kinds of antibodies and the experiment parameters were optimized. Thereby, the double antibody sandwich BA-ELISA method for the quantitative detection of CFTR protein was established. Furthermore, the reproducibility, specificity and so on were evaluated by clinical specimens of sperm.
RESULTSThe optimal concentration of coated anti-CFTR IgG was 4 µg/ml, while the biotin labeled anti-CFTR IgG was 10 µg/ml; the optimal blocking buffer was 1% BSA-PBST, the optimal time of the reaction between antigen and antibody was 60 min, the optimal chromogenic time was 15 min, the intra-assay and inter-assay coefficient were 2.16%-9.23% and 2.29%-11.71% respectively; The lowest detectable limit was 0.15 ng/ml; the standard curve had a good linear correlation of R2 = 0.962.
CONCLUSIONThe BA-ELISA method for the quantitative detection of CTFR protein is successfully established, and it is demonstrated that the method has strong specificity, high sensitivity and good reproducibility. It provides the basis and evidence of the further application of the method.
Animals ; Antibodies ; Cystic Fibrosis Transmembrane Conductance Regulator ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli ; Humans ; Peptides ; Rabbits ; Reproducibility of Results ; Sensitivity and Specificity
5.Comparative analysis of seven marine biological source of mineral drugs.
Wei SI ; Ru-na A ; Shang-rong LI ; Jing-Xian ZHANG ; Wan-ying WU ; Ya-jun CUI
China Journal of Chinese Materia Medica 2014;39(17):3321-3325
The marine biological source of mineral drugs recorded in Chinese Pharmacopoeia (2010 version) mainly including pearl, nacre, clam shell, common oyster shell, ark shell, cuttle bone, and sea-ear shell are widely used in clinical. Calcium carbonate and a small amount of protein are the main components in this type of drugs. In this paper, a systematical and comparable study were carried out by determination of calcium carbonate by EDTA titration method, the crystal of calcium carbonate by X-Ray powder diffraction and the total amino acids (TAAs) of the hydrolyzed samples by ultraviolet spectrophotometry method. As a result, the crystal structure is calcite for common oyster shell, mixture of calcite and aragonite for nacre and sea-ear shell, aragonite for the other drugs. The content of calcium carbonate ranged from 86% to 96%. Cuttle bone has the highest amount of TAAs among the seven drugs which reached 1.7% while clam shell has the lowest content of 0.16% on average. In conclusion, an effective method was developed for the quality control of marine mineral drugs by comprehensive analysis of calcium carbonate and TAAs in the seven marine mineral drugs.
Amino Acids
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analysis
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chemistry
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Animal Shells
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chemistry
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Animals
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Calcium Carbonate
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analysis
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chemistry
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Crystallization
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Edetic Acid
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chemistry
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Mollusca
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chemistry
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classification
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Pharmaceutical Preparations
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analysis
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chemistry
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standards
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Quality Control
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Reproducibility of Results
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Seawater
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Species Specificity
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Spectrophotometry, Ultraviolet
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X-Ray Diffraction
6.Shugan Yiyang Capsules for the treatment of asthenospermia: A clinical study.
Ya-lei SHI ; Min-jian ZHANG ; Wan-jun CHENG ; Yang-fan OU
National Journal of Andrology 2015;21(7):634-636
OBJECTIVETo observe the clinical efficacy of Shugan Yiyang Capsules in the treatment of asthenospermia and its action mechanisms.
METHODSWe randomly assigned 135 asthenospermia patients to groups A (n = 47), B (n = 45), and C (n = 43) to be treated with Shugan Yiyang Capsules, oral levocarnitine, or combination of the two. We observed sperm quality and the level of α-glucosidase in the seminal plasma before and after medication.
RESULTSThe total effectiveness rate was 70.21% in group A (markedly effective in 16 cases and effective in 17), 68.89% in group B (markedly effective in 15 cases and effective in 16), and 83.72% in group C (markedly effective in 16 cases and effective in 20), significantly higher in C than in A and B (P < 0.05). Both sperm quality and the level of α-glucosidase in the seminal plasma were improved in the three groups of patients, most obviously in group C.
CONCLUSIONShugan Yiyang Capsules can be used for the treatment of asthenospermia, and its effect can be enhanced in combination with oral levocarnitine.
Asthenozoospermia ; drug therapy ; enzymology ; Biomedical Research ; Capsules ; Carnitine ; administration & dosage ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Semen ; enzymology ; Spermatozoa ; alpha-Glucosidases ; analysis
7.Relations of synovial angiogenesis and PTEN/PI3K/AKT signaling pathway in rats with adjuvant arthritis.
Xiao-jun ZHANG ; Jian LIU ; Lei WAN ; Yue SUN ; Fang WANG ; Ya-jun QI ; Chuan-bing HUANG
China Journal of Orthopaedics and Traumatology 2015;28(1):71-74
OBJECTIVETo observe the change of PTEN/PI3K/AKT pathway hypoxia-inducible factor (HIF-1α), vascular endothelial growth factor (VEGF) in rats with adjuvant arthritis and to explore the mechanism of neovasculization in rheumatoid arthritis.
METHODSThirty rats were randomly divided into normal control group and model control group. The model control group were established the model of adjuvant arthritis using Freund's complete adjuvant. At 19 days after modeling, the expression of microvascular density (MVD), HIF-1α, VEGF were detected by ELISA assay and PTEN, PI3K, AKT were detected by Werstern Blotting.
RESULTSCompared with the normal control group, paw swelling, arthritic index were increased, and the expression of MVD, VEGF, HIF-1α of serum, PI3K, AKT of synovial tissue were significantly increased, PTEN was significantly decreased in model control group. PI3K, HIF-1α were positively correlated with MVD; VEGF, AKT were positively correlated with paw swelling; PTEN was negatively correlated with the arthritis index; HIF-1α was positively correlated with VEGF; PI3K was positively correlated with AKT, PTEN was negatively correlated with PI3K, AKT, VEGF.
CONCLUSIONImbalance of PTEN/PI3K/AKT pathway in rats with adjuvant arthritis is one of the mechanisms of synovial neovasculization.
Animals ; Arthritis, Experimental ; physiopathology ; Hypoxia-Inducible Factor 1, alpha Subunit ; physiology ; Neovascularization, Pathologic ; etiology ; PTEN Phosphohydrolase ; physiology ; Phosphatidylinositol 3-Kinases ; physiology ; Proto-Oncogene Proteins c-akt ; physiology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; physiology
8.Hysteroscopic treatment of women with previous cesarean scar defect.
Da-bao XU ; Ya-qiong HE ; Hui LIU ; Ya-jun WAN ; Min XUE
Journal of Southern Medical University 2010;30(2):394-396
OBJECTIVETo investigate the effects of hysteroscopic treatment of women with previous cesarean scar defect (PCSD).
METHODSFrom May 2006 to October 2008, 12 patients with PCSD were diagnosed and treated hysteroscopically in our hospital, all of them were successful followed-up for one year postoperatively, and their clinical data were analyzed.
RESULTSAll 12 hysteroscopic procedures were completed successfully, and there were no surgical complications. Nine patients with longer periods and 1 patient with intermenstrual spotting preoperatively remained asymptomatic after hysteroscopic surgery, and 1 patient with longer periods and infertility experienced normal periods, while remained infertility, and the remaining 1 patient complaining postcoital bleeding preoperatively had recurrence of the bleeding.
CONCLUSIONHysteroscopic surgery of women with PCSD was minimally invasive and effective.
Adult ; Cesarean Section ; adverse effects ; Cicatrix ; etiology ; surgery ; Female ; Follow-Up Studies ; Humans ; Hysteroscopy ; Postoperative Complications ; surgery ; Uterine Diseases ; surgery
10.Rapid detection of alpha-globin gene αααtriplets with droplet digital PCR.
Xiao-Qian GONG ; Xue-Huang YANG ; Lin-Li QIAO ; Ya-Jun CHENG ; Wan-Jun ZHOU
Journal of Southern Medical University 2017;37(9):1265-1269
OBJECTIVETo establish a rapid method for detection of alpha-globin gene αααbased on droplet digital PCR (ddPCR) technique.
METHODSThe differential sequence between the X1 and Y1 box of α1 gene was selected as the amplicon of the target gene with β-actin as the reference gene. The specific primers and TaqMan probes were designed, and then a quantitative method for detecting the copy number was established based on ddPCR technique. The sensitivity and accuracy of the method were evaluated by detecting 28 samples of known genotypes and 60 clinical samples.
RESULTSThe ddPCR-based method accurately identified the genotypes of all the 28 samples with known genotypes and detected 5 cases of αα/αααfrom the 60 clinical samples, and the results were verified by MLPA. The sensitivity and accuracy of this method were both 100% for detecting alpha-globin gene ααα.
CONCLUSIONThis ddPCR-based method for detecting αααtriplet can be applied for population screening and in routine clinical molecular diagnosis with simple operation, rapid analysis and accurate results.