1.Antibacterial and nematicidal activity of endophyte ThF-11 of Tripterygium hypoglaucum (Levl.)Hutch
Xi ZHENG ; Guohong LI ; Xin WANG ; Yan QI ; Chunping WAN
Chinese Pharmacological Bulletin 2014;(12):1709-1712,1713
Aim To investigate the chemical compo-nents and antibacterial and nematicidal activity of en-doparasitic fungi ThF-1 1 of Tripterygium hypoglaucum (Levl.)Hutch.Methods The endophyte ThF-1 1 was isolated,purified and identified based on fungal ITS sequence.The chemical components were isolated and identified with column chromatography and NMR spec-trometer,respectively.The nematicidal and antibacte-rial activity was investigated by biological active meth-od and inhibition zone method.Results The endo-phyte ThF-1 1 was identified as Penicillium simplicissi-mum,the compounds were identified as Penicillic acid [1 ],2,4-dimethoxy-6-methylphenol [2]and 5 (6)-dihydropenicillic acid [3 ] based on spectral data. Compound [1 ]showed significant activity against Cae-norhabditis elegans in a dose-dependent manner,and had antibacterial activity to Escherichia coli,Bacillus cereus,Bacillus subtilis, Staphylococcus aureus and Pseudomonas aeruginosa.Conclusion These com-pounds are firstly isolated from endophyte ThF-1 1 of T. hypoglaucum (Levl.)Hutch.Compound [1 ]shows strong nematicidal activity and antibacterial activity, and is worth further studying and developing.
2.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
3.The kinetics of in vitro lysozyme deposition on rigid gas-permeable contact lens for a short period
Jin ZHANG ; Ziyi WAN ; Kaihui NAN ; Qi ZHENG ; Jia QU ; Tingfei XI
Chinese Journal of Tissue Engineering Research 2010;14(3):473-476
BACKGROUND: The contact lenses were easily contaminated by adsorbing components from the tear film, particularly protein after wearing for a period of time. Lysozyme adsorption dynamics of fluorosilicone acrylate contact lenses has been studied in order to further improve data of protein adsorption, reduce adsorbing amount of surface protein, and prevent surface contamination of contact lenses.OBJECTIVE: To investigate the adsorption dynamics of fluorosilicone acrylate contact lenses to lysozyme in vitro. METHODS: A stock solution of lysozyme was prepared in Hanks balanced salt solution (2.0 g/L, solution Ⅰ) and different trifluoroacetic acid (TFA) concentrations were prepared. Recovery experiment, the contact lenses were placed in shaking incubator at 37 ℃ for varying time intervals. After incubation there was a single rinsing in Hanks balanced salt solution. Contact lenses in control group were placed in diluted water, and contact lenses in the other group were placed in different concentrations of TFA. For deposition, FSA contact lenses in experimental group were placed in shaking incubator at 37 ℃ for varying time intervals. After incubation there was a single rinsing in Hanks balanced salt solution. Then FSA contact lenses were immersed in 0.2% TFA solution. The amount of lysozyme was assayed with BCA method.RESULTS AND CONCLUSION: Lysozyme which attached to fluorosilicone acrylate contact lenses could be resolved by TFA, and the recovery was influenced by the immersed time and the concentration of TFA. The optimal time was 1 hour, and the optimum concentration was 0.2%. The adsorption dynamics of lysozyme on FSA contact lenses was a second-phased process, i.e., lysozyme adsorption increased rapidly during 10 minutes-1 hour, reached a plateau at 1 hour, stably adsorbed during 1-24 hours, and reached a saturation of 0.349 mg/cm~2. The recovery of lysozyme was lower at 10 and 30 minutes, but reached 90%-100% while the time of incubation was between 40 minutes and 24 hours.
4.Cytocompatibility of nano-hydroxyapatite/bacterial cellulose nanocomposites and its residues
Qi ZHENG ; Tingfei XI ; Yanmei CHEN ; Zhaoxu WANG ; Yizao WAN ; Chuan GAO
Chinese Journal of Tissue Engineering Research 2010;14(3):405-409
BACKGROUND: The nano-hydroxyapatite/bacterial cellulose (nHAP/BC) nanocomposites has a good prospect of application in bone tissue engineering, and the bone tissue engineered materials and its degradation products Should have excellent compatibility. This study further assessed DAN synthesis cycle using flow cytometry on the basis of evaluating cell compatibility by metabolic 3-(4, 5-dim ethylthiazo 1-2-y 1) -2, 5-Dipheny 1-2H-tetrazolium (MTT) assay. OBJECTIVE: To evaluate the cytocompatibility of a new-pattern nHAP/BC nanocomposites and its residues. METHODS: Effects of nHAP/BC nanocomposites and its residues on morphclogicel changes in osteoblasts were observed using in vitro cell culture method. Effects of nHAP/BC nanocomposites and its residues on osteoblast growth and prclifera'don were evaluated by MTT assay. Cell cycle phase changes were detected using flow cytometry to evaluate matsdal effects on cell proliferation on molecular levels. RESULTS AND CONCLUSION: The nHAP/BC nanocomposites and its residues had neither remarkable effects on cell morphology, nor significant inhibition on osteoblast growth and proliferation. Test of MTT cytotoxicity showed that the average cell proliferation rate was over 80% after treated with the material and its residues, with the cytotoxity grade of 1 (non-toxic). Flow cytometry indicated that the rate of G_0/G_1 was reduced, and the rates of S, G2/M were increased, and the synthesis of DNA was increased, the cellular growth and repair in osteoblasts was accelerated. These indicated that nHAP/BC nanocomposites have good cytocompatibility, and it will be safe and prospected scaffolds in bone tissue engineering.
5.Exploring mechanism of cardiac function decreasing in patients with ankylosing spondylitis based on Keap1-Nrf2-ARE signaling pathway
Yajun QI ; Jian LIU ; Li ZHENG ; Lei WAN ; Yunxiang CAO ; Yue SUN ; Fang WANG
Chinese Journal of Immunology 2014;(5):654-661
Objective:To explore the mechanism of cardiac function decreasing in patients with ankylosing spondylitis based on Keap1-Nrf2-ARE signaling pathway .Methods:Experiment group included 140 ankylosing spondylitis ( AS) patients and control group included 60 healthy individuals .Echocardiography ( UCG) was used to detect cardiac function parameters .Enzyme-linked immunosor-bent assay(ELISA) was adopted to determine serum proteins ,oxidative stress indicators and cytokines .Erythrocyte sedimentation and plasma C reactive protein ,immunoglobulin were detected with Westergren method and Hitachi 7060 automatic biochemical analyzer re-spectively.Results:(1)Compared with the normal control group , character of AS group in active stage:cardiac function parameters(E peak,EF,E/A),antioxidant indicators(SOD,CAT,TAOC),cytokines(IL-4,IL-10)decreased (P<0.01 or P<0.05 all).A peak, Keap1,Nrf2,oxidation index(ROS,RNS,MDA),cytokines(IL-1,TNF-α),inflammatory indicators(ESR,CRP)increased(P<0.01 or P<0.05 all).(2)Compared with the normal control group, character of AS group in stable stage: cardiac function parameters(E peak,E/A),Keap1,antioxidant indicators(SOD,CAT,TAOC),cytokines(IL-4,IL-10)decreased (P<0.01 or P<0.05 all).A peak, Nrf2,oxidation index(ROS,RNS,MDA),cytokines(TNF-α),inflammatory indicators(ESR,CRP) increased(P<0.01 or P<0.05 all).(3)Compared with AS group in stable stage , character of AS group in active stage:cardiac function parameters(E peak,EF,E/A),antioxidant indicators(SOD,TAOC),cytokines(IL-10)decreased (P<0.01 or P<0.05 all).Keap1,Nrf2,oxidation index(ROS, RNS,MDA),cytokines(IL-1β,TNF-α),inflammatory indicators(ESR,CRP),clinical evaluation indicators (VAS,BASDAI,BASFI, BASMI,BAS-G)increased(P<0.01 or P<0.05 all).(4)Compared with AS patients with normal Keap1 or Nrf2 of peripheral blood, A peak increased while E peak and E/A decreased in AS patients with abnormal Keap1 or Nrf2 of peripheral blood (P<0.01 or P<0.05).(5)Spearman correlation analysis showed:AS parameters of cardiac function in patients with E peak and antioxidant indicators (SOD,CAT,TAOC),cytokines(IL-4,IL-10)showed significant positive correlation.E peak and Keap1, Nrf2,oxidation index(ROS, RNS,MDA),cytokines(IL-1β,TNF-α),inflammatory indicators(ESR,CRP),clinical evaluation indicators (VAS,BASDAI,BASFI, BASMI,BAS-G),IgG,course of the disease were significantly negatively correlated .A peak and Nrf2,oxidation index ( ROS,RNS, MDA) ,cytokines ( IL-1β,TNF-α) ,clinical evaluation indicators , Palpitation showed significant positive correlation .A peak and antiox-idant indicators(SOD,CAT,TAOC),cytokines(IL-4,IL-10)were significantly negatively correlated .E/A and Keap1,antioxidant indi-cators(SOD,CAT,TAOC),cytokines(IL-4,IL-10)showed significant positive correlation.E/A and oxidation index(ROS,RNS, MDA) ,cytokines ( TNF-α) ,cxcinical evaluation indicators ,course of the disease were significantly negatively correlated .EF and course of the disease were significantly negatively correlated .FS and Keap1 showed significant negative correlation .Conclusion:Cardiac func-tion decreasing in patients with ankylosing spondylitis is 33.56%.The characteristic performance is that E peak , E/A decreased while A peak increased.E peak,E/A and antioxidant indicators ,anti-inflammatory cytokines show significant positive correlation .E peak,E/A and Keap1,oxidation index,pro-inflammatory cytokines,inflammatory indicators are significantly negatively correlated .A peak and Keap1,oxidation index ,pro-inflammatory cytokines show significant positive correlation .A peak and antioxidant indicators ,anti-inflam-matory cytokines are significantly negatively correlated .The rising of Keap1 expression cause activate the Keap 1-Nrf2-ARE signaling pathway .After the signaling pathway is activated ,the body's antioxidant ability decreased , pro-inflammatory cytokines imbalanced , in-flammation index increased .When the body is long be in unbalance condition , the number of abnormal immune complex deposition in-creased.In the end,the result is that AS is caused and cardiac function is decreased .
6.Change of Capillary Pericapillary Cells in Rats with Myocardial Infarction and Effect of Supplementing Qi and Activating Blood Circulation Herbs
Kun HUANG ; Dandan YANG ; Shuwen GUO ; Qing SUN ; Lu ZHANG ; Xin QI ; Ting WAN ; Chenglong ZHENG
Chinese Journal of Information on Traditional Chinese Medicine 2013;(7):38-41,42
Objective To observe the change of capillary pericapillary cells in rats with myocardial infarction and the influence of supplementing qi and activating blood circulation herbs, and explore its mechanism of improving myocardial perfusion. Methods The rat model was established by ligaturing the left anterior descending coronary artery. On the base of ECG evaluation, successfully modeled rats were randomly divided into the model group, group treated with supplementing qi and activating blood circulation Chinese medicine (activating blood and supplementing qi group), group treated with Perindopril (Perindopril group), group treated with Tongxinluo Capsules (Tongxinluo group). The sham-operation group was taken as the control. There were totally 5 groups. The model group and the sham-operation group were treated with normal saline. The changes of myocardial capillary density (MCD) and number of pericapillary cells on the 7th, 28th day after medicinal administration were observed. Results On the 7th and 28th day, the MCD decreased significantly and the number of capillary pericapillary cells increased significantly in the model group compared with the sham-operation group (P<0.01). The MCD increased significantly in the activating blood and supplementing qi group, Perindopril group and Tongxinluo group compared with the model group (P<0.01). The number of pericapillary cells decreased significantly in the activating blood and supplementing qi group, Perindopril group and Tongxinluo group compared with the model group (P<0.01). Conclusion The supplementing qi and activating blood circulation herbs can improve regional myocardial blood supply by decreasing the number of pericytes and promoting regeneration of capillary.
7.CT and MRI features of dermatofibrosarcoma protuberans
Xiaotao ZHENG ; Qi WAN ; Xinchun LI ; Qiao ZOU ; Yingshi DENG ; Yudong YU ; Yingying BAO ; Qiang LEI
Chinese Journal of Medical Imaging Technology 2017;33(4):586-589
Objective To investigate CT and MRI features of dermatofibrosarcoma protuberans (DFSP).Methods Totally 16 patients with DFSP confirmed by pathology were enrolled.Tumor morphology,CT and MRI imaging appearance (11 cases underwent plain and enhanced CT,5 cases underwent plain and enhanced MRI) were analyzed retrospectively.Results DFSP usually occurred in the skin of truck,head and neck,protruding from the skin surface in different extent.Some lesions even suspended out of the skin.The lesions were divided into nodular type (n=12) and diffuse type (n=4) according to their morphological appearance.The tumors usually demonstrated as iso-density or slightly low density solid mass compared to muscle on CT.On MRI,it usually demonstrated as low signal on T1WI and high signal on T2WI.Tumor blood supply was rich,and it usually showed progressively moderate to strong enhancement.The signs within DFSP include hanging sign (n=2),skin tail sign (n=6),fascia tail sign (n=l),fat tail sign (n=4).Conclusion DFSP can be characterized by nodular or diffuse lesions,the manifestations of different form are slightly different,but still have a certain characteristic.
8.Effects of ING5 gene on the malignant phenotype of breast cancer Bcap-37 cells
Yang SONG ; Yizeng WAN ; Shupeng ZHAO ; Fengjie QI ; Lei FANG ; Jicheng WU ; Shuai SHI ; Huachuan ZHENG
Medical Journal of Chinese People's Liberation Army 2017;42(1):12-16
Objective To investigate the effects of inhibitor of growth 5 (ING5) gene on the proliferation, apoptosis, migration and cell cycle of human breast cancer Bcap-37 cells.Methods The eukaryotic ING5-expressing plasmid and GFP-empty plasmid were steadily transfected in Bcap-37 cells, the expression of green fluorescent protein was measured with fluorescence microscopy, and the high expression of ING5 was measured by real time-PCR. Bcap-37-ING5 cells served as the experimental group, Bcap-37-GFP cells as the mock group and Bcap-37 as the control group. The effects of ING5 on the proliferation were detected by MTT, the cell cycle and apoptosis were detected by Flow cytometry, and the cell migration was detected by cell wound scratch assay and Transwell experiment.Results Bcap-37 cell lines steadily expressing ING5 protein with GFP-tag were acquired by stable transfection. ING5 over-expression inhibited the proliferation and led to G2 arrest of Bcap-37 cells, increased cells apoptosis and decreased the cell migration ability (P<0.05).Conclusion ING5 over-expression may have reverse effect for malignant phenotype of breast cancer cells, and may be employed to indicate the biomarker of prognosis of breast cancer patients and regarded as a target of gene therapy.
9.Carbon monoxide inhalation protects against limb ischemia/reperfusion injury.
Jing LI ; Zhen-long ZHU ; Wan-zheng QI ; Zhong-li SHI
Chinese Journal of Applied Physiology 2010;26(1):33-36
OBJECTIVETo study the protective effect of carbon monoxide(CO) inhalation in the limb ischemia/reperfusion (I/R) injury of rats.
METHODSForty-four Sprague-Dawley rats were randomly divided into three groups: S, I/R and RC groups. I/R injury model was made by the occlusion of the femoral artery for four hours and the reperfusion for forty-eight hours. RC group was exposed to medical air mixed CO (the volume fraction of CO: 0.05%) during limb reperfusion in rats. The other two groups were exposed to the routine air. Gross morphology of the ischemic limb, wet-to-dry weight ratio (W/D), and skeletal muscle histopathology were observed. The apoptosis index and expression levels of Bax and Bcl-2 in the muscle were assessed with Flow Cytometry. The activities of serum lactate dehydrogenase (LDH) and creatine kinase (CK) were tested by Automatic Biochemical Analyzer.
RESULTSCompared to I/R group, W/D, serum LDH and CK activities, the apoptosis index and Bax expression level in the muscle were all significantly decreased, the Bcl-2 expression level was significantly increased, gross morphology of the ischemic limb and muscle histopathology were obviously improved in RC group.
CONCLUSIONInhaling exogenous CO can attenuate limb I/R injury.
Administration, Inhalation ; Animals ; Carbon Monoxide ; pharmacology ; Creatine Kinase ; metabolism ; Extremities ; blood supply ; Femoral Artery ; Ischemia ; physiopathology ; L-Lactate Dehydrogenase ; metabolism ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; bcl-2-Associated X Protein ; metabolism
10.Effects of carbon monoxide inhalation on severe limb ischemia/reperfusion injury and its damages lead to shock.
Jin ZHAO ; Zhong-Li SHI ; Wan-Zheng QI ; Zhen-Long ZHU
Chinese Journal of Applied Physiology 2011;27(2):149-153
OBJECTIVETo study the protective effect of carbon monoxide (CO) inhalation on the serious limb ischemia/reperfusion (I/R) injury, and which effects caused to shock in rats.
METHODS36 SD rats were randomly divided into I/R, I/R + CO (RC), sham operation (S) groups. I/R injury models were made by the occlusion of the femoral artery for 8 h and the reperfusion for 12 h, 10 d. Before reperfusion of 2 h, RC group started to breathe medical air containing CO (the volume fraction of CO: 0.075%) continuously, until after reperfusion for 4 h, a total of inhalation 6 h. S, I/R groups exposed to air, breathe freely. Caudal artery pressures (CAP), ten days survival rate, serum lactate dehydrogenase (LDH) and creatine kinase (CK) activity, limb wet - to - dry weight ratio (W/D) and the pathologic changes of limb were observed.
RESULTSOnce the reperfusion started, the CAP decreased rapidly in I/R group, and the mean reduced to(5.3259 +/- 0.3832) kPa when reperfusion for 8 h. Compared to I/R group, the CAP decreased slower and smaller in RC group, moreover, its mean reduced to (8.3300 +/- 0.4224) kPa when reperfusion for 8 h. The 10 d survival rate in I/R group was that 8 rats died all between reperfusion for 13 - 20 h. Only 1 rat died in RC group and the other 7 rats were still alive when reperfusion for 10 d. Compared to I/R group, the pathological features of the ischemic limb were significant ly improved, and the figures of W/D, serum LDH and CK value were remarkable lower in RC group (P < 0.05).
CONCLUSIONInhaling exogenous low-dose CO has a reverse regulation in the blood pressure decline caused by serious limb I/R injury in rats. And at the same time, it can effectively prevent the occurrence of shock, reduce physical damage, significantly increase the survival rate of animals.
Administration, Inhalation ; Animals ; Carbon Monoxide ; administration & dosage ; pharmacology ; Creatine Kinase ; blood ; Extremities ; blood supply ; L-Lactate Dehydrogenase ; blood ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; physiopathology ; prevention & control ; Shock ; etiology ; prevention & control