3.Expression of cytokines and suppressor of cytokine signaling in peripheral blood mononuclear cells derived dendritic cells from primary biliary cirrhosis patients
Yunheng ZHOU ; Hongxia MA ; Guangya CAO ; Guanghua RONG ; Renqian ZHONG
Chinese Journal of Laboratory Medicine 2012;35(3):216-220
Objective To study the role of suppressor of cytokine signaling ( SOCS ) in the pathogenesis of primary biliary cirrhosis( PBC),the levels of SOCS protein and the changes of function of dendritic cell(DC) were respectively observed from PBC patients.MethodsThe study population consisted of 10 patients of PBC and 8 healthy controls.Phenotypic analysis of cultured peripheral blood mononuclear cells (PBMC)-derived DC was performed by flow cytometry (FCM),such as CD83,CD86 and human leukocyte antigen DR (HLA-DR).The levels of interleukin-10 (IL-10),interferon-γ( IFN-γ) and IL-12 in culture supematant of DC were measured by enzyme linked immunosorbent assay ( ELISA ).The protein levels of SOCS1 and SOCS3 were detected by Western blot ( WB ).The features of changes in these parameters were analyzed between the two groups.ResultsThe expression of CD83,CD86 and HLA-DR in PBC patients were ( 79.4 ± 4.8 ) %,( 86.5 ± 6.3 ) % and (90.0 ± 3.5 ) %,which were significantly higher than those in healthy control group[ (68.3 ±4.1 )%,(74.2 ±6.3)% and (83.6 ±7.6)% ],respectively (t =5.340,4.120,2.514,P <0.05).The levels of IL-12 and IFN-γin PBC patients were (53.5 ± 11.1)and (32.0 ±9.0) ng/L,which were significantly higher than those in healthy control group[ (32.1 ± 10.7) and (15.4 ± 8.1 ) ng/L; t =4.123,3.818,P < 0.01 ].There were not any significant difference of IL-10 level between PBC patients [ (7.0 ± 4.6) ng/L ] and the healthy controls [ ( 5.8 ± 4.2) ng/L; t =0.563,P > 0.05 ].The proteins levels of SOCS1 and SOCS3 in PBMC-derived DC from PBC group were decreased significantly than those in healthy control group.ConclusionsThe results suggest that the PBMC-derived DC in PBC patients has greater ability of potent maturation and antigen presentation function.The decreased expression of SOCS levels may be associated with the excessive immunological reaction and the breakdown of self-tolerance.
4.Research on the effect of protection against ventilator-induced lung injury via regulation of caveolin-1/heme oxygenase-1 signaling
Rong ZHONG ; Jun XIAO ; Zhihui YU ; Ji ZHOU ; Chunguang DAI
Chinese Critical Care Medicine 2015;(7):568-573
ObjectiveTo determine whether the inhibition of caveolin-1 tyrosine residues 14 (Cav-1-Y14) phosphorylation with protein tyrosine kinase inhibitors (PP2) will upregulate heme oxygenase-1 (HO-1) activity to protect against ventilation induced lung injury in vivo of an animal model.Methods Fifty-four male Sprague-Dawley (SD) rats were randomly divided into nine groups (eachn = 6). Group A served as normal control group, in which rats did not receive ventilation but tracheotomy. Groups B1 and B2 received lung protective ventilation respectively for 1 hour or 2 hours. Groups C1 and C2 received high tidal volume (40 mL/kg) ventilation for 1 hour or 2 hours, respectively. The group D1 or D2 also received high tidal volume ventilation for 1 hour or 2 hour respectively, but they were given PP2 1 hour before high tidal volume ventilation. The groups E1 and E2 also received high tidal volume ventilation respectively for 1 hour or 2 hours, but tyrosine kinase inhibitor PP2 and HO-1 inhibitor zinc protoporphyrinⅨ(ZnPPⅨ) were given to animals 18 hours before high tidal volume ventilation. All the animals were sacrificed after ventilation, and the specimens of lung tissues and bronchoalveolar lavage fluid (BALF) were harvested. Then the changes in pathology of lung tissue was observed, and diffuse alveolar damage scores (DAD) were calculated, myeloperoxidase (MPO) activity was measured by colorimetric analysis, lung wet/dry ratio (W/D) was estimated. The expressions of phosphorylated caveolin-1 (P-Cav-1-Y14), caveolin-1 (Cav-1) and HO-1 were determined by Western Blot. The expressions of high mobility group B1 (HMGB1) and advanced glycation end product receptor (RAGE) in lung tissues were assayed with immunohistochemistry staining. The levels of tumor necrosis factor-α(TNF-α) in BALF were measured by enzyme linked immunosorbent assay (ELISA).Results There was no significant difference in all the parameters between group A and groups B. Compared with group B1, DAD score, W/D ratio, the activity of MPO and the concentration of TNF-α in BALF in group C1 were significantly increased [DAD score:7.97±0.59 vs. 0.55±0.13, W/D ratio: 5.70±1.61 vs. 5.04±0.63, MPO (U/g): 1.82±0.14 vs. 0.77±0.26, TNF-α(ng/L): 370.10±29.61 vs. 54.38±8.18, allP< 0.05], and the injury in ventilation 2 hours group was more serious than that in ventilation 1 hour group. Compared with groups C, all the parameters in groups D were significantly decreased. The parameters in groups E were significantly higher than those in groups A, B, and D, but no significant difference was found as compared with groups C. Compared with groups B, the protein expressions of Cav-1 and P-Cav-1-Y14 (gray value) in groups C were significantly increased (1 hour: 1.49±0.02 vs. 1.26±0.13, 1.34±0.02 vs. 0.87±0.04;2 hours: 1.58±0.02 vs. 1.27±0.27, 1.31±0.01 vs. 0.95±0.02, allP< 0.05), and the expression of HO-1 protein (gray value) was significantly decreased (1 hour: 0.59±0.02 vs. 1.10±0.01, 2 hours: 0.49±0.01 vs. 1.20±0.02, both P< 0.05). No significant difference in Cav-1 protein expression between groups D as well as groups E and groups C. The protein expression of P-Cav-1-Y14 in groups D and E was significantly lower than that in groups C. The protein expression of HO-1 in groups D was significantly higher than that in groups C, but the phenomenon was not found in groups E as compared with groups C. Compared with group A, the positive expression of HMGB1 and RAGE in lung tissue in groups C and E was significantly increased, but no significant difference was found between groups B as well as groups D and group A.Conclusion Cav-1-Y14 phosphorylation is the key factor for ventilator induced lung injury, which can not only lead to a decrease in vascular barrier function, but also inhibit the activity of HO-1 enzyme, thus further aggravates inflammatory injury of the lung as induced by mechanical ventilation.
5.Regulation of Nrf2 pathway to protect ventilator induce lung injury in vivo via inhibition of caveolin ;phosphorylation
Rong ZHONG ; Jun XIAO ; Chunguang DAI ; Zhihui YU ; Ji ZHOU
Chinese Critical Care Medicine 2016;28(6):547-552
Objective To investigate whether the inhibition of caveolin-1 (Cav-1) phosphorylation will regulate effectively nuclear factor-erythroid 2-related factor (Nrf2) signal pathway and downstream effector molecules and protest against ventilation induced lung injury (VILI) in an animal model in vivo. Methods Ninety male Sprague-Dawley (SD) rats were randomly divided into nine groups (each n = 10): sham group in which rats did not receive ventilation but received tracheotomy; lung protective ventilation (PV) for 1 hour or 2 hours group; mechanical ventilation (MV) at high volume tidal (VT, 40 mL/kg) for 1 hour or 2 hours group; protein tyrosine kinase inhibitor PP2 or rosiglitazone (Rsg) pretreatment + high VT ventilation for 1 hour or 2 hours groups. The two pretreatment groups were given intraperitoneal injection PP2 15 mg/kg or intragastric administration of Rsg 5 mg/kg 1 hour before ventilation respectively. The rats were sacrificed after model reproduction, and bronchoalveolar lavage fluid (BALF) was collected. Pulmonary vascular permeability was measured by Evans blue (EB). The levels of tumor necrosis factor-α (TNF-α), activator protein-1 (AP-1), nuclear factor-κB (NF-κB), and interleukin-8 (IL-8) in BALF were determined by enzyme linked immunosorbent assay (ELISA). Then the lung tissues were collected, the lung wet/dry ratio (W/D) was calculated, the changes in pathology was observed with light microscope, and myeloperoxidase (MPO) activity was determined by colorimetric analysis. Nrf2 mRNA was determined by reverse transcription-polymerase chain reaction (RT-PCR). The expressions of Cav-1 tyrosine residues 14 phosphorylation (pCav-1-Y14), Cav-1, peroxisome proliferators-activated receptor γ (PPARγ) and claudin-5 as well as Nrf2 in cytoplasm and nucleus were determined by Western Blot. The positive expressions of PPARγ and claudin-5 in lung tissues were assayed with immunohistochemistry staining. Results There were no obvious pathological changes in the lung tissue in sham group and PV groups, and there were no significant differences in all the parameters between the two groups either. However, the injury in lung tissue was severe in the high VT groups in which W/D ratio, EB contents, MPO activity, and TNF-α, AP-1, IL-8, NF-κB levels in BALF as well as the protein expressions of Cav-1 and pCav-1-Y14 were significantly higher than those of sham group and PV groups, and the protein expressions of PPARγ and claudin-5 were significant lower than those of sham group and PV groups with a dose-dependent manner; but Nrf2 expressions in cytoplasm and nucleus did not show a statistical increase. After pretreatment of PP2 or Rsg, W/D ratio, MPO activity, EB contents, TNF-α, AP-1, IL-8, and NF-κB in BALF were significantly decreased as compared with those of high VT group, and RT-PCR showed significant up-regulation of Nrf2 mRNA in lung tissues too. Moreover, there was a statistically significant increase in expressed Nrf2 proteins in nucleus in PP2 or Rsg groups as compared with those of high VT groups [Nrf2 in nucleus (gray value): 0.61±0.06, 0.56±0.06 vs. 0.31±0.02 at 1 hour, 0.38±0.06, 0.43±0.07 vs. 0.22±0.03 at 2 hours; all P < 0.05], but no significant difference was found in the expression of Nrf2 protein in the cytoplasm among all groups. The protein expressions of pCav-1-Y14 in PP2 pretreatment groups were significantly lower than those of high VT groups (gray value: 0.89±0.04 vs. 1.48±0.02 at 1 hour, 0.86±0.02 vs. 1.31±0.01 at 2 hours; both P < 0.05); but expressed PPARγ proteins and expressed claudin-5 proteins in PP2 or Rsg pretreatment groups were significantly higher than those of high VT groups [PPARγ (gray value): 0.34±0.07, 0.42±0.13 vs. 0.17±0.07 at 1 hour, 0.38±0.09, 0.33±0.07 vs. 0.16±0.03 at 2 hours; claudin-5 (gray value): 0.33±0.05, 0.38±0.07 vs. 0.14±0.03 at 1 hour; 0.30±0.06, 0.31±0.04 vs. 0.17±0.04 at 2 hours; all P < 0.05]. Conclusions The inhibition of Cav-1-Y14 phosphorylation can increase the expression of Nrf2 in the nucleus, then result in an increase in the protein expressions of PPARγ and claudin-5 of its effector molecules. This effect can reduce the inflammation and capillary permeability of lung tissue in the model of VILI.
6.Analysis of varieties and standards of labiatae medicinal plants used in Tibetan medicine.
Jun-wei HE ; Lan CAO ; Hua-rong ZHOU ; Wei-hong ZHONG ; Guo-yue ZHONG
China Journal of Chinese Materia Medica 2015;40(7):1419-1424
In this paper, an analysis was made on the varieties and standards of labiatae medicinal plants used in Tibetan medicine. The results showed 71 species of labiatae plants in 21 genera (including varieties) recorded in relevant literatures, involving 44 varieties of medicinal materials. Specifically, seven species (9.9%) were intersected with traditional Chinese medicines (TCM), 19 varieties (43%) were recorded in Chinese medicinal material standards at all levels, and 27 species (38%) were source plants. In Tibetan medicine standards and literatures, there are great differences between Tibetan names and translated Chinese names and among varieties of source plants. Apart from a few of varieties intersected with traditional Chinese medicines had complete standards and regulations in Chinese Pharmacopoeia, most of species only had characters, microscopic, physical and chemical identifications in Standards Issued by Ministry of Health-Tibetan Medicine, Tibetan Medicine Standard and local standards. Therefore, the Tibetan medicinal material variety-source specification and quality standard system shall be promoted on the basis of literatures research, investigations for resources and current applications and modern pharmaceutical studies.
Drugs, Chinese Herbal
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chemistry
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pharmacology
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Humans
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Lamiaceae
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chemistry
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classification
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Medicine, Tibetan Traditional
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standards
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Phytotherapy
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standards
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Plants, Medicinal
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chemistry
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classification
7.Alterations and clinical significance of peripheral blood T lymphocyte subsets in patients with septic acute kidney injury
Rong TANG ; Yong ZHONG ; Zhou XIAO ; Ting MENG ; Xiaozhao LI ; Xiangcheng XIAO ; Qiaoling ZHOU
Journal of Chinese Physician 2015;(3):324-327
Objective To investigate the alterations of peripheral blood T lymphocyte subsets in patients with sepsis and septic acute kidney injury, and explore the clinical significance.Methods Fifty-five patients with sepsis and forty-three patients with septic acute kidney injury were enrolled in this study. At the same period, thirty healthy subjects were enrolled as the control group.T lymphocyte subsets inclu-ding CD3 +T, CD4 +T, CD8 +T cells, and CD4 +T/CD8 +T in peripheral blood were detected by flow cy-tometry, and acute physiology and chronic health evaluation Ⅱ( APACHE Ⅱ) were graded within twenty-four hours after admission.Then, correlation of the APACHEⅡscores and T lymphocyte subsets was ana-lyzed.Results In the septic acute kidney injury group, peripheral blood CD3 +T, CD4 +T cell percenta-ges, and CD4 +T/CD8 +T ratio were significantly lower than those in the control group and the sepsis group (all P <0.05).In the septic acute kidney injury group with stage 3, CD3+T, CD4 +T cell percentages, and CD4 +T/CD8 +T ratio in the patients were significantly lower than those in stage 1 and stage 2 ( all P <0.05).In the septic acute kidney injury group,CD3 +T, CD4 +T cell percentages, and CD4 +T/CD8 +T ra-tio were significantly lower in dead group than those in survival group (all P <0.05).APACHEⅡscores in patients with sepsis were significantly negatively correlated with peripheral blood CD4 +T cell percentages and CD4 +T/CD8 +T ratio ( r =-0.645,-0.492, allP <0.05).Conclusions There are varying de-grees of cellular immune imbalance in patients with sepsis and septic acute kidney injury, characterized by decline of circulating CD3 +T, CD4 +T cell percentages, and CD4 +T/CD8 +T ratio.CD4 +T cell percenta-ges and CD4 +T/CD8 +T ratio are closely related to the severity and prognosis of septic acute kidney injury.
9.Clinical application of anterolateral femoral skin flap pedicled with high site cutaneous branches of the transverse branch of lateral circumflex femoral artery
Pei-Ji WANG ; Qi-Rong DONG ; Jian-Zhong QIN ; Kai-Long ZHOU ; Bo JIANG ; Xian-Zhong ZHANG
Chinese Journal of Microsurgery 2000;0(02):-
Objective To report the clinical effect of free transplanting for soft tissue defects pedieled with high site cutaneous branches of the transverse branch of lateral cirumflex femoral artery.Methods Cu- taneous branches of the descending branch of lateral circumflex femoral artery were found small or abscent in 7 patients.The anterolateral femoral skin flap was pedicled with high site cutaneous branches of the transverse branch to repair the soft tissue defects of the arm,hand,leg and foot,rather than with the descending ones. The size of the flap ranged from 15 cm?6 cm to 28 cm?13 cm,with part muscle valve,iliotibal tract and lat- eral femoral cutaneous nerve.The fractures were performed with internal or external fixation.Results All of the anterolateral femoral skin flap survived well postoperatively in the 7 cases and had good appearance and sensation at one stage.The function of the repaired extremities recovered well.Conclusion The anterolat- eral femoral skin flap pedicled with high site cutaneous branches of the transuvrse branch of lateral circumflex femoral artery has many advantages of good blood supply and large size.The flap was secluding,and can be taken with some muscle and lateral femoral cutaneous nerve.When cutaneous branches of the descending branch of lateral circumflex femoral artery is small or abscent,the anterolateral femoral skin flap with high site cutaneous branches of the transverse branch of lateral circumflex femoral artery is an optimal alternative.
10.Can PDA patient in early stage of Eisenmenger syndrome accept interventional surgery ?
Xiaoke SHANG ; Liang ZHONG ; Gangcheng ZHANG ; Ting PENG ; Dingyang LI ; Rong LU ; Xiaoxian DENG ; Hongmei ZHOU
Chinese Journal of Interventional Cardiology 2014;(9):553-557
Objective To explore whether the PDA patients in the early stage of Eisenmenger syndrome can accept interventional surgery. Methods Three patients were choose from the “National Multicenter Prospective Registration Study on Pulmonary Artery Hypertension”, Clinical assessment, as well as examinations and registrations associated with PAH, was conducted in accordance with relevant provisions of the registration study within 1 wk prior to surgery. The way of the interventional treatment were right heart catheterization and pulmonary vasodilator testing (a capsule of iloprost solution for inhalation), and measurement of the pulmonary arterial pressure, descending aortic pressure, ratio of pulmonary to systemic blood flow, ratio of pulmonary to systemic blood pressure, pulmonary capillary wedge pressure [mean≤15 mmHg(1 mmHg=0.133 kPa)], before and after testing. After clear the results of pulmonary vasodilator test all patients underwent transcatheter closure testing, if it was positive,the amplatzer was released if her family members’ approved;Conversely, the amplatzer was quickly withdrawn with negative results, and symptomatic treatment was conducted based on the patient’s clinical symptoms. Results The first cases pulmonary vasodilator test was negative, but positive in transcatheter closure testing. Satisfaction immediate effect of surgery, and the occluder was released with her family members’ approval. patients of Example 2 and Example 3 were negative in pulmonary vasodilator testing and transcatheter closure testing, can not released the occluder. Conclusions Some Patents Ductus Arteriosus (PDA) patients in early stage of Eisenmenger syndrome is feasible to accept interventional therapy , try plugging test is may be an important indicator of prognostic assessment.