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Aim To study the effects of extracellular acidosis on articular chondrocytes pyroptosis and its possible mechanisms. Methods Primary articular chondrocytes were incubated in different pH and NAC. The expression of proinflammatory cytokines IL-1β, IL-18, ASC, NLRP3, caspase-1 were detected by Western blot and real-time PCR. The state of pyropto-sis was identified by AO/EB staining and LDH con-tents. The expression of ROS was observed by DCFH-DA, and ELISA was used to detect the IL-1β,IL-18 in cultured supernatants. Results Compared with the normal cell, extracellular acidosis could increase the expression of IL-1β, IL-18, ASC, NLRP3 and caspase-1 , upregulate the fluorescence intensity of in-tercellular ROS, accompanied with the promoted release of LDH. Moreover, it is observed that extra-cellular acidosis could also induce chondrocytes death by AO/EB staining. NAC,the scavenger of ROS could inhibit these effects of extracellular acidosis on chon-drocytes. Conclusion Extracellular acidosis may in-duce chondrocyte pyroptosis via upregulating the intra-cellular ROS content.
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Aim To explore the proteomics mechanism of the differentiation induction effect of 4-amino-2-trif-luoromethyl-phenyl retinate(ATPR)on human leukemi-a K562 cells. Methods Human leukemia K562 cells were incubated with the same concentration (1 × 10 - 6 mol·L - 1 ) of ATPR or ATRA for 48 hours. The total cell proteins were collected, purified and digested by trypsin, solid phase extraction, and the peptides were detected by ESI-LC-MS / MS. The difference of the pro-tein expression between the cells treated with ATPR and ATRA was compared by using the Discoverer Pro-teome 1. 2 software, and the molecular function, the biological process and other information of those pro-teins were analyzed based on the DAVID, KEGG, STRING databases. Results 120 specific proteins were identified only in the ATPR group, 143 only in the ATRA group, and 422 other proteins in both groups. Results of DAVID analysis showed that ATPR-induced specific proteins were mainly involved in 39 biological processes of proteins and macromolecules metabolism, protein transport and localization and so on. Results of KEGG analysis revealed that ATPR-in-duced proteins participated in signal pathways, mainly metabolic pathways, PI3K-Akt signal pathway, TGF-beta signal pathway and other pathways in cancer. String protein interaction network analysis displayed that ATPR-induced proteins, like EIF3A, EIF6, RPL3, RPL8, RPL13, RPL7A, RPL21, RPS3, RPS14, NACA, BTF3, NHP2L1, PPP2CA proteins had direct interactions with more than or equal to 10 associated proteins. Conclusion The differentiation induction effect of ATPR on K562 cells might be as-cribed to the ATPR-induced proteins interaction net-work and the specific central proteins it induced, which are involved in the regulation of cell prolifera-tion, differentiation and apoptosis.
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Nesfatin-1, discovered in 2006 by Oh-I as an 82-ami-no-acid peptide derived from the precursor protein nucleobindin2 (NUCB2), has been identified to play an important role in the regulation of food intake and energy metabolism. Recently, it has also been found that Nesfatin-1 might be associated with the pathogenesis of depression. This article reviewed the advances in related studies on Nesfatin-1 at home and abroad, which should throw light in expliciting the physiological function of Nesfatin-1 and understanding the neurobiological mechanism of depression.
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Aim To investigate the effect of 4-Amino- 2-Trifluoromethyl-Phenyl Retinate on human breast cancer cells MDA-MB-231 and the possible mecha-nisms. Method Human breast cancer MDA-MB-231 cells were incubated with different concentrations of ATPR in vitro. MTT assay was performed to measure the proliferation of MDA-MB-231 . Cell growth curves were made by counting cells and morphologic changes were observed by Wright-Giemsa staining. The differ-entiation marker mucin-1 ( MUC-1 ) was measured by enzyme linked immunosorbent assay ( ELISA ) . Cell cycle was examined by Flow cytometry ( FCM ) . The expression of retinoic acid receptors ( RARs) and reti-noid X receptors ( RXRs ) were detected by Western blot and Quantitative real-time PCR (q-RT-PCR),re-spectively. Results Compared with solvent group, ATPR could inhibit the proliferation of MDA-MB-231 cells in a time-and dose dependent manner and induce the maturing and normality of morphology. The express of MUC-1 was significantly decreased, and the progres of cell cycle was blocked in the G0/G1-phase. The ex-pression of RARγ was decreased. Conclusions AT-PR could inhibit proliferation and induce differention of MDA-MB-231cells, it′s associated with RARγ.
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OBJECTIVE:To provide empirical reference for protecting the supply of clinical drugs. METHODS:Literature re-view was combined with the conditions of drugs that lost the bidding in the centralized bidding in Anhui province to determine the short drugs need to investigate. Questionnaire was adopted to investigate the situations and reasons of short drugs in 5 third-grade class-A hospitals of 5 areas in Anhui province from Nov. 2013 to Oct. 2014. RESULTS:A total of 5 questionnaires were sent out, and 5 were received with effective response rate of 100%. There were totally 54 short drugs in the 5 third-grade class-A hospitals, including the most serious shortage of drugs for neurocirculatory system,accounting for 20.37%. Shortage was mainly due to the low price of drugs,accounting for 48.15%,and insufficient supply,less suffering patients/low dosage and other reasons. CON-CLUSIONS:In view of the shortage of drugs,government departments should improve the drug pricing and bidding policy,pro-duction enterprises should enhance the enterprise production and development capabilities,business companies should optimize the distribution pattern of network and medical institutions should establish drug management system with a clear division of power and responsibility to relieve the drug shortages.
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Aim To observe the effect of BAPTA-AM on extracellular acid-induced autophagy in rat articular chondrocytes and its possible mechanisms.Methods Rat articular chondrocytes were isolated from Sprague-Dawley rats and incubated with different pH medium. The states of autophagy were examined by acridine or-ange (AO ) staining .Moreover,the expressions of LC3 ,Beclin-1 ,ULK1 ,CaMKKβ,AMPK and mTOR were detected using Western blot or quantitative real-time PCR (qRT-PCR ). Intracellular calcium ([Ca2+]i )was analyzed by a Ca2+-imaging method. Results Compared with pH 6.0 group,BAPTA-AM could significantly decrease the activation of autophagyinduced by acid exposure,and the expressions of autophagy markers including LC3 Ⅱ,Beclin1 and ULK1were also decreased,accompanied with reduced acidinduced [Ca2 +]i influx,decreased proteins expressionof CaMKKβand phosphorylatedAMPK,and increasedphosphorylation of mTOR.Conclusion BAPTAAMcan significantly restrain acidinduced autophagy in ratarticular chondrocytes,the mechanism of which may beassociated with decreased Ca2 + influx.
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Aim TostudytheroleofASIC1aonthe matrix turnover and MAPK expression of the rat articu-lar chondrocytes with extracellular acidosis. Methods ArticularchondrocyteswereisolatedfromSprague-Dawley rats, and their phenotype was determined by toluidine blue and immunocytochemical staining. The GAG content of cell culture supernatant was deter-mined by dimethyl-methylene blue spectrophotometric assay, while Hyp content by chloramine T assay. ELISA assay was used to measure MMP-2 , TIMP-2 content. Furthermore, the ERK1/2, p38 MAPK phos-phorylation protein expression levels were tested by Westernblotassay.Results ASIC1acontributedto the effect of GAG, Hyp and TIMP-2 levels reduction induced by extracellular acidification, while the effect of MMP-2 was weaker. Moreover, ASIC1a could in-crease the ERK1/2 , p38 MAPK phosphorylation pro-teinexpressionlevels.Conclusion ASIC1acould regulate rat articular chondrocytes matrix turnover via ERK1/2 and p38 MAPK signaling pathway, and there-by inhibit the rat articular cartilage damage induced by acidosis.
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Aim To develop a sensitive,specific and accurate method for quantifying a novel derivate of all-trans-retinoic acid, 4-amino-2-trifluoromethyl-phenyl retinate (ATPR)in rat tissues to investigate the tissue distribution of ATPR in rats.Methods Sprague-Daw-ley (SD)rats were killed by exsanguination at 2,4,7 h after a single intragastric administration with one dose of ATPR (20 mg·kg-1 )or at 5 min,1 h,5 h after a single intravenous administration with one dose of AT-PR (7 mg·kg-1 ).The concentration of ATPR in the tissues was determined by high performance liquid chromatography (HPLC)method.Results After the rats were administrated intragastrically, the highest concentration of ATPR was observed in intestine,fol-lowed by liver,spleen and lung,while the distribution in heart,kidney,fat and brain was very little.Howev-er,the highest concentration of ATPR was in liver after given intravenously,followed by spleen and lung,and very low in heart,kidney,intestines,fat and brain. Conclusion The distribution of ATPR is higher in liv-er after administrated both intragastrically and intrave-nously,suggesting the potential anti-proliferation and differentiation inducing effects of ATPR targeting at liv-er cancer.
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Objective To develop an LC-MS/MS method for determining the concentration of wogonin in dog plasma and investigate the pharmacokinetics and bioavailability by different administrations of wogonin in Beagle's dogs. Methods LC-MS/MS was employed in determining the concentration of wogonin with the selected ion monitoring model after liquid-liquid extraction with ethyl acetate of dog plasma samples. The lower limit of quantification was 0.105 μg/L. Target ions were at m/z 285.0→270.0 for wogonin and 373.3→305.3 for finasteride. In a randomized, self-control, and cross-over study, six male Beagle's dogs were treated with different administration methods in three test periods. Pharmacokinetic parameters were calculated with DAS software (Ver. 2.0). Results The calibration curve was linear in the range of 0.105-107.36 μg/L for wogonin in dog plasma samples. The main pharmacokinetic parameters of ig administration (native drug of 15 mg/kg and solution preparation of 5 mg/kg) and iv route were as follows: Cmax (2.5 ± 1.1), (7.9 ± 3.3), and (6838.7 ± 1322.1) μg/L, tmax (0.7 ± 0.3) and (0.3 ± 0.2) h for the both former, AUC0-1 (7.1 ± 2.0), (21.0 ± 3.2), and (629.7 ±111.8) μg·h/L. The absolute bioavailability of native and solution of wogonin were (0.59 ± 0.35)% and (3.65 ± 2.00)%, respectively. Conclusion The validated method is convenient, sensitive, and specific, and the improvement of wogonin solubility could remarkably increase the absolute bioavailability.
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Objective The fingerprint method based on high-performance liquid chromatography for the quality control of bidens bipinnata L were firstly described and optimized. Methods HPLC system was used to obtain the fingerprint spectrum of bidens bipinnata L. The chromatography condition was ODS C18 column (4. 6mm×255mm,25μm),mobile phase Methanol-0. 2% HAc,and flow speed 1. 0ml ? min-1, detection wavelength 363nm.Results In this experiment,the standard fingerprint had been established and the similitude degree of 10 batches bidens bipinnata L. has also been obtained(0.941～0.999). Conclusion The method was accurate,reliable,good reproducibility, strong characteristic and could be used for the standard of general judgement and the quality estimation about the bidens bipinnata L.
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Acid-sensing ion channels(ASICs) are a novel class of ligand-gated cation channels activated by extracellular acidification and belong to the epithelial sodium channels(DEG/ENaC) superfamily.Their biological functions have recently been found not only in the central nervous system but also relevant to the physiology and pathology of non-neuronal tissues such as taste buds, cardiovascular system and bones.This review concerns the latest research on the expression and functions of ASICs in non-neuronal tissues so as to promote the understanding of their physiological and pathological functions.
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Aim To investigate the proliferation of HSCs stimulated by exogenous TGF-β1 (transforming growth factor betal),observe the effect of TFB(total flavonoids of Bidens Bipinnata L.)on smad2/7,typeⅠcollagen mRNA and protein expression of HSCs and study the protective effect and molecular mechanism of TFB on hepatic fibrosis.Methods HSCs were isolated with collagenase Ⅳ perfusion in situ and density gradient centrifugation. The effect of TFB on cell proliferation was observed by MTT colormetric assay. The auto-secretion of TGF-β1 and synthesis of type Ⅰ collagen were measured by enzyme-linked immuneadsordent assay (ELISA).Moreover,the expression of smad2/7, typeⅠcollagen mRNA and protein was measured by semi-quantitative RT-PCR and Western blot methods respectively.Results TFB could markedly inhibit the proliferation of HSCs of liver fibrosis rats stimulated by TGF-β1 and production of TGF-β1 and type Ⅰ collagen.In addition,TFB treatment could significantly down-regulate smad2 and type Ⅰ collagen mRNA expression and up-regulated smad7 mRNA expression of HSCs Smad2 protein expression of HSCs stimulated by TGF-β1 was also down-regulated by TFB.Conclusion TFB has the protective effect against hepatic fibrosis by inhibiting the activation of TGF-β1 signaling pathway and suppressing the HSC proliferation.
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Objective To study the expression and significance of acid-sensing ion channels(ASICs)in rat articular cartilage with adjuvant arthritis. Methods Complete Freund's adjuvant(CFA) was prepared by suspending heat-killed Bacillus Calmette Guerin(BCG) in liquid paraffin at 10 mg/ml. CFA-induced arthritis was developed by injection of 100 μl CFA emulsion intradermally into the right hind paw. The morphological changes of articular tissues was observed by light microscope; RT-PCR and immunoblotting analyses were used to detect ASICs in rat articular cartilage with adjuvant arthritis. Results RT-PCR and western blot showed that ASIC1a, ASIC2a and ASIC3 were present in the articular cartilage of normal and model group, the ASICs mRNA levels in the model group were higher than in the normal group detected by semiquantitative analysis (P＜0.01), ASICs protein levels in model group were higher than those in the normal group (P＜0.01) when examined by immunoblotting. Conclusion The results show that the expression of ASICs in AA articular cartilage is enhanced and it may be related with articular cartilage breakdown.
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Objective:To investigate the influencing factors and medication compliance of hypertension.Method: 3 grade-Ⅲand 15 grade-Ⅱgeneral hospitals in Anhui Province were selected by random sampling.732 outpatients with a hypertension course of 6 months or above were interviewed during July and October of 2004 with the medication compliance schedule.All data were input into computers with the software Epi info 6.04,and analyzed with SPSS 11.5.Result:①The medication compliance schedule had high reliability and validity,which could be used in the surveys of medication compliance of hypertension.②The medication compliance rate of the 732 patients was only 46.4%.③Trust in medical pro- fessionals,more attention paid to the disease,more knowledge of non-compliance harm,satisfactory family income,and longer consulting time were the common advancing factors in improving the medication compliance,but a longer duration of the medication regimen,a larger number of preparations and drugs,and higher frequency of taking medicine were the bloc- king factors.Conclusion:The medication compliance of hypertensives was generally not good,and it was influenced by many factors.It is necessary that medical professionals,patients,families and society all take part in its improvement.
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Objectives To study the effect of total flavones of Bidens pilosa L (TFB) on cytokines production in liver fibrosis rats. Methods Rat liver fibrosis was induced by CCl450%, 0.1 ml?(100 g)-1 body weight twice a week for 18 weeks. TFB (160,80,40 mg?kg-1) was used daily via gastrogavage at 9 week. Levels of TNF-? and IL-1? in serum were determinate by radioimmunoassay. Liver samples were collected after experiments and stained by immuninochemistry of TGF-?1 and NF-?B. Moreover TGF-?1 mRNA expression in liver tissue was detected by RT-PCR technology. Results TFB (160, 80 mg?kg-1) could significantly reduce serum TNF-? and IL-1? contents; TFB(160, 80, 40 mg?kg-1) could effectively prevent the expression of NF-?B,as was TGF-?1 of TFB(160, 80 mg?kg-1). Moreover TFB (160, 80 mg?kg-1) could significantly reduce TGF-?1 mRNA in liver fibrosis rats. Conclusion TFB had protective effect on liver fibrosis by its inhibition of cytokine production.
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Aim To explore the effect of 4-amino-2-trifluoromethyl-phenyl retinate(ATPR)on proliferation,differentiation activity in K562 cell line,and to research the mechanisms.Methods Cell proliferation was assessed by MTT assay.Cell differentiation index was analyzed by NBT reduction test.Morphologic changes were observed by Wright's staining in the light microscope. Cell cycle was determined by FCM.The mRNA expression of Cyclin D1,Cyclin E,CDK2,CDK4,CDK6,P21cip1,P27kip1,P57kip2,PCNA mRNA were detected by RT-PCR.While the protein expression of cyclin D1 and CDK4 was detected by Western blot.Results The growth of K562 cells was inhibited in a dose-dependent manner.NBT reduction test indicated that the ATPR could induce differentiation of K562 cells and increase the positive cell ratio.Morphologic changes were observed after Wright's staining using inverted phase contrast microscope.The proportion of cells in G0/G1 phase increased while S phase cells decreased.Cell cycle progression was blocked in the G1 phase.The expression of Cyclin E,cyclin D1,CDK2,CDK4,CDK6 mRNA decreased,while PCNA,P21 cip1,P27 kip1 change was not obvious,but P57 (kip2) mRNA expression was increased.Cyclin D1 and CDK4 protein expressions were reduced as well.Conclusions ATPR inhibits the growth of K562 cells and induces differentiation.P57 kip2 plays a key role in differentiation.Moreover,high level of P57kip2 is regulated via inhibiting its degradation through reducing proteasome-dependent proteolysis,and ATPR plays a role in cell cycle arrest.
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Aim To explore the therapeutic effect of total flavones of Bidens Bipinnata L (TFB) on liver fibrosis in rats and its mechanism. Methods The model of rat liver fibrosis was adopted which was induced by CCl4 injection. The effects of TFB were observed on the levels of serum HA,PCⅢ,CIV and Hyp in rats liver fibrosis,and on liver histopathological changes as well as collagen hyperplasia formation in liver tissue. The apoptosis of HSC were detected by double-staining of ?-smooth muscle actin (?-SMA) and TUNEL. The study in vitro was carried out on the culture of isolated hepatic stellate cells. Cell proliferation was detected with MTT assay. Cell apoptosis was detected by electron microscopy and flow cytometry. Results TFB can significantly reduce serum HA,PCⅢ,CⅣ and Hyp contents in liver fibrosis of rats,improve the liver pathologic injury,reduce collagen hyperplasia in liver of liver fibrosis rats,inhibit the activation and proliferation of HSC,and promote the apoptosis of HSC. In addition TFB could significantly inhibit the proliferation and increased the apoptosis of isolated and cultured HSC compared with the control group. Conclusions TFB has a significant therapeutic effect on the liver fibrosis rats,probably its inhibition of proliferation and stimulation of activated HSC apoptosis may be an important mechanism of its therapeutical effect against liver fibrosis.
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OBJECTIVE:To explore a reimbursement mechanism for adverse drug reactions(ADR)suitable to the condition of China.METHODS:Based on the condition in China,we evaluated the feasibility of applying insurance mechanism into ADR reimbursement from aspects of the development of insurance industry and pharmaceutical industry as well as the characteristics of ADR etc.Meanwhile we put forward some suggestions on the construction and operation of the ADR liability insurance.RESULTS & CONCLUSIONS:ADR liability insurance can help guarantee patients' legitimate rights of life health,settle medical disputes,facilitate the development of pharmaceutical industry and promote ADR reporting and management work.To carry out ADR liability insurance,issues like insurance rate,national policy support and evolving of ADR monitor network should be taken into consideration.
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AIM To observe the effects of rhIL 10 on psoriatic models and immune function in mice. METHODS The anti psoriatic effect of rhIL 10 was studied by using the models of vaginal epithelium and tail scales of mouse. ConA induced proliferation and production of IL 2 and IFN ? by T lymphocytes, LPS induced production of IL 1 and TNF ? by the peritoneal macrophages and the activity of NK cells, LPS induced proliferation of B lymphocyte were determined. RESULTS The result showed that rhIL 10 (5, 20 and 80 ?g?kg -1 ?d -1 ,sc) could significantly inhibit the mitosis of vaginal epithelial cells in mice treated with estrogen. No abnormal changes of nucleolus and organelle were found under electronic microscope. rhIL 10 facilitates development of granular cell layers in mice tail scales. There were for 2 to 3 granular cell layers and bigger granular cells in scales under electronic microscope. CONCLUSION The anti psoriatic effect of rhIL 10 is related to inhibiting hyperplasia of epidermal cells and promoting the formation of granular cells and possessing anti inflammatory and immunomodulatory activity.
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Neovascularization is one of the early pathologic changes of rheumatoid arthritis(RA)synovium and is the major players in the promotion and perpetuation of pannus.Endostatin has antiangiogenic effect via inhibition of endothelial cell proliferation,migration and induction of apoptosis.The inhibition of angiogenesis in synovium by endostatin appears to be a promising means for the future treatment of RA.Also,endostatin has a therapeutic effect on RA by the inhibition of proliferation and induction of apoptosis in fibroblast-like synoviocytes.