Perinatal complications,such as asphyxia,can cause brain injuries that are often associated with subsequent neurological deficits,such as cerebral palsy or mental retardation.The mechanisms of perinatal brain injury a...Perinatal complications,such as asphyxia,can cause brain injuries that are often associated with subsequent neurological deficits,such as cerebral palsy or mental retardation.The mechanisms of perinatal brain injury are not fully understood,but mitochondria play a prominent role not only due to their central function in metabolism but also because many proteins with apoptosis-related functions are located in the mitochondrion.Among these proteins,apoptosis-inducing factor has already been shown to be an important factor involved in neuronal cell death upon hypoxia-ischemia,but a better understanding of the mechanisms behind these processes is required for the development of more effective treatments during the early stages of perinatal brain injury.In this review,we focus on the molecular mechanisms of hypoxic-ischemic encephalopathy,specifically on the importance of apoptosis-inducing factor.The relevance of apoptosis-inducing factor is based not only because it participates in the caspase-independent apoptotic pathway but also because it plays a crucial role in mitochondrial energetic functionality,especially with regard to the maintenance of electron transport during oxidative phosphorylation and in oxidative stress,acting as a free radical scavenger.We also discuss all the different apoptosis-inducing factor isoforms discovered,focusing especially on apoptosis-inducing factor 2,which is only expressed in the brain and the functions of which are starting now to be clarified.Finally,we summarized the interaction of apoptosis-inducing factor with several proteins that are crucial for both apoptosis-inducing factor functions(prosurvival and pro-apoptotic)and that are highly important in order to develop promising therapeutic targets for improving outcomes after perinatal brain injury.展开更多
The higher frequency of varicocele in men with infertility has drawn attention and resulted in increased research at the molecular level towards treatments. The aim of this study was to investigate the role of tumor n...The higher frequency of varicocele in men with infertility has drawn attention and resulted in increased research at the molecular level towards treatments. The aim of this study was to investigate the role of tumor necrosis factor （TNF）-related apoptosis-inducing ligand （TRAIL） and its receptors in varicocele-induced testicular dysfunction in an experimental rat model. The rats were divided into three groups： control, sham and varicocele. Varicoceles in rats were induced by partial ligation of the left renal vein and left testes. The rats were analyzed 13 weeks after surgery. The degree of DNA fragmentation within cells in the testis was determined using terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling （TUNEL） assay. Tubule degeneration was evaluated using the Johnsen score. The expression of TRAIL and its receptors was detected by immunohistochemical and Western blotting techniques. The apoptotic index, Johnsen score and the expression of TRAIL and TRAIL receptors were examined. The data are presented as the mean-.＋s.d, and were analyzed using computer software. The KruskaI-Wallis and Dunn＇s multiple comparison tests were used in the statistical analyses. The germ cell apoptotic index was increased in rats with varicoceles when compared with the sham and control groups （P=0.0031）. The Johnsen score was significantly decreased in the varicocele group when compared with the sham and control groups （P〈O.O001）. Immunohistochemical and Western blotting analyses showed that after varicocele induction, the expression of TRAIL-R1 and TRAIL-R4 in germ cells was increased and the expression of TRAIL-R2 was decreased. There are no significant differences among the groups in terms of TRAIL and TRAIL-R3 receptor expression. The results of this study indicate that TRAIL and its receptors may have a potential role in the pathogenesis of varicocele-induced testicular dysfunction.展开更多
Objective To investigate whether desferoxamine （DFO） preconditioning can induce tolerance against cerebral ischemia and its effect on the expression of hypoxia inducible factor 1 α （HIF- 1α） and erythropoietin ...Objective To investigate whether desferoxamine （DFO） preconditioning can induce tolerance against cerebral ischemia and its effect on the expression of hypoxia inducible factor 1 α （HIF- 1α） and erythropoietin （EPO） in vivo and in vitro. Methods Rat model of cerebral ischemia was established by middle cerebral artery occlusion with or without DFO administration. Infarct size was examined by TTC staining, and the neurological severity score was evaluated according to published method. Cortical neurons were cultured under ischemia stress which was mimicked by oxygen-glucose deprivation （OGD）, and the neuron damage was assessed by MTT assay. Immunofluorescent staining was employed to detect the expressions of HIF-1 and EPO. Results The protective effect induced by DFO （decreasing the infarction volume and ameliorating the neurological function） appeared at 2 d after administration ofDFO （post-DFO）, lasted until 7 d and disappeared at 14 d （P 〈 0.05）; the most effective action was observed at 3 d post-DFO. DFO induced tolerance of cultured neurons against OGD： neuronal viability was increased 23%, 34%, 40%, 48% and 56% at 8 h, 12 h, 24 h, 36 h, and 48 h, respectively, post-DFO （P 〈 0.05）. Immunofluorescent staining found that HIF-1 α and EPO were upregulated in the neurons of rat brain at 3 d and 7 d post-DFO; increase of HIF-1 α and EPO appeared in cultured cortex neurons at 36 h and 48 h post-DFO. Conclusion DFO induced tolerance against focal cerebral ischemia in rats, and exerted protective effect on OGD cultured cortical neurons. DFO significant induced the expression of HIF- 1 α and EPO both in vivo and in vitro. DFO preconditioning can protect against cerebral ischemia, which may be associated with the synthesis of HIF- 1 α and EPO.展开更多
AIM:To study the effects of Helicobacter pylori(H. pylori)tumor necrosis factor-α(TNF)inducing protein (Tip-α)on cytokine expression and its mechanism. METHODS:We cloned Tip-αfrom the H.pylori strain 26695,transfor...AIM:To study the effects of Helicobacter pylori(H. pylori)tumor necrosis factor-α(TNF)inducing protein (Tip-α)on cytokine expression and its mechanism. METHODS:We cloned Tip-αfrom the H.pylori strain 26695,transformed Escherichia coli with an expression plasmid,and then confirmed the expression product by Western blotting.Using different concentrations of Tip-αthat affected SGC7901 and GES-1 cells at different times,we assessed cytokine levels using enzyme-linked immunosorbent assay.We blocked SGC7901 cells with pyrrolidine dithiocarbamate(PDTC),a specific inhibitor of nuclear factorκB(NF-κB).We then detected interleukin(IL)-1βand TNF-αlevels in SGC7901 cells. RESULTS:Western blot analysis using an anti-Tip-α antibody revealed a 23-kDa protein,which indicated that recombinant Tip-αprotein was recombined successfully.The levels of IL-1β,IL-8 and TNF-αwere sig-nificantly higher following Tip-αinterference,whether GES-1 cells or SGC-7901 cells were used(P<0.05).However,the levels of cytokines(including IL-1β,IL-8 and TNF-α)secreted by SGC-7901 cells were greater than those secreted by GES-1 cells following treatment with Tip-αat the same concentration and for the same duration(P<0.05).After blocking NF-κB with PDTC, the cells(GES-1 cells and SGC-7901 cells)underwent interference with Tip-α.We found that IL-1βand TNF-αlevels were significantly decreased compared to cells that only underwent Tip-αinterference(P<0.05). CONCLUSION:Tip-αplays an important role in cyto-kine expression through NF-κB.展开更多
OBJECTIVE:To examine the efficacy of Qinghuayin(清化饮,QHY)in rat chronic atrophic gastritis(CAG)models and explored the molecular mechanism of QHY in treating CAG.METHODS:In total,65 Wistar rats were randomly divided...OBJECTIVE:To examine the efficacy of Qinghuayin(清化饮,QHY)in rat chronic atrophic gastritis(CAG)models and explored the molecular mechanism of QHY in treating CAG.METHODS:In total,65 Wistar rats were randomly divided into the control(n=10)and CAG groups(n=55).CAG model rats were further divided into five groups:model(n=10),vitacoenzyme(n=10),low-dose QHY(n=10),medium-dose QHY(n=10),and high-dose QHY groups(n=10).We analyzed histopathological changes using hematoxylin and eosin staining and measured interleukin(IL)-6 and IL-8 levels in serum using enzyme-linked immunosorbent assay(ELISA)(Boster Bio,Pleasanton,USA).In addition,gastrin(GAS),pepsinogen I(PGI),and PGII expressions were evaluated using ELISA.The protein and m RNA expression of toll-like receptor 4(TLR4)and toll or interleukin-1 receptor domaincontaining adaptor inducing interferon-β(TRIF)was detected by Western blotting and quantitative reverse transcription-polymerase chain reaction,respectively.RESULTS:Our results revealed that histopathological changes in CAG model rates could be restored by low-,medium-,and high-dose QHY.The changes in GAS and PGI/II expression demonstrated that QHY improved CAG.Serum IL-6 and IL-levels were decreased by QHY administration.TLR4 and TRIF were upregulated at the m RNA and protein levels in the model group but downregulated by QHY administration.CONCLUSION:We concluded that QHY could effectively improve the histopathological changes of the gastric mucosa induced by CAG in rats.The therapeutic mechanism of QHY may be related to inhibition of the inflammatory factors IL-6 and IL-8 and suppression of TLR4/TRIF m RNA and protein expression.展开更多
Acupuncture has been broadly used as a major therapeutic tool for at least 2,100 years in East Asia. Acupuncture is used for improving the pathological condition of remote organs, such as stomach, liver, brain, etc. b...Acupuncture has been broadly used as a major therapeutic tool for at least 2,100 years in East Asia. Acupuncture is used for improving the pathological condition of remote organs, such as stomach, liver, brain, etc. by stimulating skin and muscle with fine metal needles through meridians which have been regarded as pathways of Qi within the body （Ernst, 2006）. Physicians in East Asia have tried to treat many kinds of diseases by acupuncture and it was considerably success- ful. Various diseases for which acupuncture is known to be effective, are neurological diseases, including sequelae of stroke, Alzheimer＇s disease （AD）, Parkinson＇s disease （PD）, and so on （Nam et al., 2013）.展开更多
Immunoglobulin G4-related sialadenitis(IgG4-RS)is an immune-mediated fibro-inflammatory disease and the pathogenesis is still not fully understood.The aim of this study was to explore the role and mechanism of interle...Immunoglobulin G4-related sialadenitis(IgG4-RS)is an immune-mediated fibro-inflammatory disease and the pathogenesis is still not fully understood.The aim of this study was to explore the role and mechanism of interleukin-13(IL-13)in the cellular senescence during the progress of IgG4-RS.We found that the expression of IL-13 and IL-13 receptorα1(IL-13Rα1)as well as the number of senescent cells were significantly higher in the submandibular glands(SMGs)of IgG4-RS patients.IL-13 directly induced senescence as shown by the elevated activity of senescence-associatedβ-galactosidase(SA-β-gal),the decreased cell proliferation,and the upregulation of senescence markers(p53 and p16)and senescence-associated secretory phenotype(SASP)factors(IL-1βand IL-6)in SMG-C6 cells.Mechanistically,IL-13 increased the level of phosphorylated signal transducer and activator of transcription 6(p-STAT6)and mitochondrial-reactive oxygen species(mt ROS),while decreased the mitochondrial membrane potential,ATP level,and the expression and activity of superoxide dismutase 2(SOD2).Notably,the IL-13-induced cellular senescence and mitochondrial dysfunction could be inhibited by pretreatment with either STAT6 inhibitor AS1517499 or mitochondria-targeted ROS scavenger Mito TEMPO.Moreover,IL-13 increased the interaction between p-STAT6 and c AMP-response element binding protein(CREB)-binding protein(CBP)and decreased the transcriptional activity of CREB on SOD2.Taken together,our findings revealed a critical role of IL-13 in the induction of salivary gland epithelial cell senescence through the elevated mitochondrial oxidative stress in a STAT6–CREB–SOD2-dependent pathway in IgG4-RS.展开更多
OBJECTIVE:To observe the effects of moxibustion at bilateral Feishu(BL13)and Xinshu(BL15)combined with benazepril on myocardial cells apoptosis index,the expression levels of apoptosis-related proteins cytochrome c(Cy...OBJECTIVE:To observe the effects of moxibustion at bilateral Feishu(BL13)and Xinshu(BL15)combined with benazepril on myocardial cells apoptosis index,the expression levels of apoptosis-related proteins cytochrome c(Cyt-C)and apoptosis-inducing factor(AIF)in chronic heart failure(CHF)rats.METHODS:Sixty-five rats were randomly divided into normal group(n=10)and model-I group(n=55).After modeling,CHF rats in model-I group were divided into model group,moxibustion group,benazepril group,moxibustion plus benazepril group(abbreviated as aibei group,the same below),10 rats in each group.Echocardiogram index was examined by echocardiography.Hemodynamic indices were measured by rat cardiac function meter.Serum B-type brain natriuretic peptide(BNP)was detected by enzymelinked immunosorbent assay.Myocardial cells apoptosis index was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling staining.Pathological changes of myocardial tissues were observed by hematoxylin and eosin staining.The expression levels of Cyt-C and AIF in myocardial tissues were detected by Western blot.RESULTS:Compared with normal group,ejection fraction and left ventricular diameter shortening rate in model-Ⅰgroup were significantly reduced,myocardial cells of rats in model group exhibited unclear transverse striations,cells swellings and vacuoles,cardiac functions were deteriorated,serum BNP level,myocardial cells apoptosis index,and the expression levels of Cyt-C and AIF were significantly increased.Compared with model group,myocardial cells of rats in moxibustion group,benazepril group,and aibei group were dyed more evenly,muscle fibers were arranged relatively neatly,cardiac functions were improved,serum BNP level,myocardial cells apoptosis index,and the expression levels of Cyt-C and AIF were significantly decreased.Compared with aibei group,cardiac functions were worsened,myocardial cells apoptosis index,and the expression levels of Cyt-C and AIF were increased.CONCLUSION:Moxibustion at bilateral Feishu(BL13)and Xinshu(BL15)combined with benazepril could improve CHF better than moxibustion at bilateral Feishu(BL13)and Xinshu(BL15)or benazepril alone.The mechanisms might be that they can inhibit the expressions of Cyt-C and AIF,and inhibit the apoptosis of cardiomyocytes.展开更多
The corneal crosslinking （CXL） with riboflavin and ultraviolet-A （UVA） is a new therapy method to successfully treat infectious keratitis in clinical practice. However, there are rare reports on the complications ...The corneal crosslinking （CXL） with riboflavin and ultraviolet-A （UVA） is a new therapy method to successfully treat infectious keratitis in clinical practice. However, there are rare reports on the complications of CXL such as the secondary keratitis. The diverse clinical outcomes on keratitis have highlighted the necessity to further evaluate the efficacy and complications of CXL. We reviewed the positive and negative reports on UVA/riboflavin related with keratitis and provided our opinion on the therapeutic and side effect of UVA/riboflavin crosslinking on keratitis.展开更多
Programmed cell death （PCD） signaling pathways are import- ant contributors to acute neurological insults such as hypox- ic-ischemic brain damage, traumatic brain injury, stroke etc. The pathogenesis of all these di...Programmed cell death （PCD） signaling pathways are import- ant contributors to acute neurological insults such as hypox- ic-ischemic brain damage, traumatic brain injury, stroke etc. The pathogenesis of all these diseases is closely linked with ab- erration of apoptotic cell death pathways. Mitochondria play a crucial role during PCD, acting as both sensors of death signals, and as initiators of biochemical path- ways, which cause cell death （Bras et al., 2005）. Cytochrome c was the firstly identified apoptogenic factor released from mitochondria into the cytosol, where it induces apoptosome formation through the activation of caspases. Other proteins, such as apoptosis inducing factor （AIF）, have been subsequently identified as mitochondrial released factors. AIF contributes to apoptotic nuclear DNA damage （Bras et al., 2005）. in a caspase-independent way展开更多
It was found that the growth of malignant tumour in mice was inhibited and the ability of immune cell’s dissolving cancer cells was enhanced by ultralow frequency (ULF) pulsed gradient magnetic field. The DNA content...It was found that the growth of malignant tumour in mice was inhibited and the ability of immune cell’s dissolving cancer cells was enhanced by ultralow frequency (ULF) pulsed gradient magnetic field. The DNA contents of nuclei decreased which indicated that magnetic field can block DNA replication and mitosis of cancer cells. It was observed that magnetic field inhibited the cancer cell’s metabolism, lowered its malignancy, and restrained its rapid and heteromorphic growth. The morphology properties of Programmed Cell Death (PCD) of the cancer cells of the treated group by magnetic field was observed for the first time. The heterochromatin condensed and coagulated together along the nuclear membrane; the endoplasmic reticulums expanded and fused with the cellular membrane; many apoptotic bodies which were packed by the cellular membrane appeared and were devoured by the lymphocytes and plasma.展开更多
Since the welded constructions produce easily stress corrosion cracking (SCC) or fatigue disruption in corrosive medium or under ripple load, two methods inducing compressive stress on structural surfaces by anti-weld...Since the welded constructions produce easily stress corrosion cracking (SCC) or fatigue disruption in corrosive medium or under ripple load, two methods inducing compressive stress on structural surfaces by anti-welding-heating treatment (AWHT) and explosion treatment (ET) are presented. The results show that they are good ways to resisting SCC on the welded SS400 steel or other components.展开更多
During our 4-year work in The Republic of Yemen, the authors used acupuncture therapy to restore consciousness and induce resuscitation plus scalp acupuncture in 80 cases of acute apoplexy, with 63 cases cured, accoun...During our 4-year work in The Republic of Yemen, the authors used acupuncture therapy to restore consciousness and induce resuscitation plus scalp acupuncture in 80 cases of acute apoplexy, with 63 cases cured, accounting for 78.75%. This therapy is obviously superior to the traditional principle of point selection in acupuncture used in 25 cases as to curative effect, treatment course and other aspects. There is an obvious statistical difference (P【 0.005).展开更多
The effect of formed CH_3NH_3 at the heterojunction on properties of CH_3NH_3PbI_3 material is investigated based on experiment and theoretical calculation. Our calculation results show that the giant dielectric const...The effect of formed CH_3NH_3 at the heterojunction on properties of CH_3NH_3PbI_3 material is investigated based on experiment and theoretical calculation. Our calculation results show that the giant dielectric constant, anomalous hysteresis and long-lasting polarization for CH_3NH_3PbI_3 originate from the formed CH_3NH_3 at the heterojunction. It is found that the induced weak EPS by the reorientation of CH_3NH_3 sub-group along the built-in electric field enables us to effectively increase the ordering of entire lead-halide framework. In addition, the heterojunction has an advantage of channel separation between carrier transport and electron diffusion. These properties of the heterojunction are the main origin of the high efficiency of CH_3NH_3PbI_3 solar cells.展开更多
BACKGROUND： Previous studies have reported that statins are less toxic to the human body and have greater antitumor activity; however, few studies have addressed the antitumor effect of statins combined with tumor ne...BACKGROUND： Previous studies have reported that statins are less toxic to the human body and have greater antitumor activity; however, few studies have addressed the antitumor effect of statins combined with tumor necrosis factor-related apoptosis inducing ligand （TRAIL）. OBJECTIVE： To explore the effect of TRAIL combined with mevastatin on the proliferation and apoptotic cell death of a human glioma cell line SWO-38, and to study its mechanism of action. DESIGN, TIME AND SETTING： An in vitro control experiment was performed at the Central Laboratory of the Third Hospital Affiliated to Sun Yat-sen University, between January and April 2009. MATERIALS： The human SWO-38 cell line was provided by Cell Research, Department of Animal Experimental Center of Sun Yat-sen University; human recombinant soluble TRAIL by R＆D, USA; and mevastatin by Sigma, USA. METHODS： SWO-38 cells were separately incubated in TRAIL （100, 200, 300, 400, and 500 tJg/L） and mevastatin （5, 10, 20, 30, and 40 pmol/L） for 72 hours. In addition, SWO-38 cells were incubated in TRAIL （300 μg/L）, mevastatin （30 μmol/L）, and a solution containing both TRAIL and mevastatin for 12, 24, 48 and 72 hours. MAIN OUTCOME MEASURES： Cell proliferation was detected using methyl thiazolyl tetrazolium assay; cell apoptosis was observed using Hoechst 33258 staining and fluorescence microscopy and was measured using Annexin V/propidium iodide flow cytometry; TRAIL R1/DR4 and TRAIL R2/DR5 protein expressions levels were measured using indirect immunofluorescence staining combined with flow cytometry in the recombinant soluble TRAIL （rsTRAIL, 300 tJg/L）, mevastatin （30 IJmol/L） and combination groups; TRAIL R1/DR4 and TRAIL R2/DR5 mRNA expression was detected using real-time polymerase chain reaction. RESULTS： rsTRAIL, mevastatin and their combination inhibited tumor proliferation in a time- and dose-dependent manner. The proliferation inhibitory rate and apoptosis rate of human SWO-38 cells in the combined group were significantly greater than the rsTRAIL or mevastatin alone group （P 〈 0.01）. TRAIL R1/DR4 and TRAIL R2/DR5 protein and mRNA expressions were increased in the combination group compared with mevastatin or rsTRAIL alone after 72 hours （P 〈 0.01）. CONCLUSION： Both rsTRAIL and mevastatin inhibit the proliferation and apoptosis of the human glioma cell line SWO-38, while their combination enhances the anti-tumor effect. The mechanism of action possibly correlates to the upregulation of TRAIL R1/DR4 and TRAIL R2/DR5 mRNA expression by mevastatin, thereby enhancing the cell sensitivity to rsTRAIL.展开更多
BACKGROUND： Receptors for tumor necrosis factor related apoptosis inducing ligand （TRAIL） include death receptor 4, death receptor 5, decoy receptor 1, and decoy receptor 2. Activation of death receptor 4 and 5 sel...BACKGROUND： Receptors for tumor necrosis factor related apoptosis inducing ligand （TRAIL） include death receptor 4, death receptor 5, decoy receptor 1, and decoy receptor 2. Activation of death receptor 4 and 5 selectively kills tumor cells. OBJECTIVE： To detect TRAIL receptor expression in glioblastoma by immunohistochemistry and RT-PCR, and to compare this expression to that in normal brain tissue. DESIGN： Observational analysis. SETTING： Department of Pathology, the First Affiliated Hospital of Zhengzhou University; Henan Tumor Pathology Key Laboratory. PARTICIPANTS： Twenty-five patients （17 males and 8 females） who received glioblastoma resection were selected from the Fifth Affiliated Hospital of Zhengzhou University, between September 2003 to June 2004. All glioblastoma samples were diagnosed pathologically. Twenty patients （12 males and 8 females） with craniocerebral injury who received normal brain tissue resection were selected in the same time period. There were no significant differences in sex and age between glioblastoma patients or between craniocerebral injury patients （P 〉 0.05）. All patients and appropriate relatives provided informed consent, and this study was approved by the local research ethics committee. METHODS： Polyclonal antibody against TRAIL receptors and an immunohistochemical kit （batch number： 200502） were purchased from Boster Company, Wuhan. Immunohistochemistry： Expression of death receptor 4, death receptor 5, decoy receptor l, and decoy receptor 2 were observed in both glioblastoma and normal brain tissue. The experiment was performed according to the kit instructions, and positive staining was brown-yellow. Assessment： There were no positive signals （-）; weakly positive signals, positive cells 〈 25% （＋）; weakly positive signals, positive cells 25%-50% （＋＋）; strongly positive signals, positive cells 50%-75% （＋＋＋）; strongly positive signals, positive cells 〉 75% （＋＋＋＋）. Evaluation： Expression levels of TRAIL receptors were estimated in both normal brain tissue and glioblastoma. Expression of decoy receptor 1 and decoy receptor 2 mRNA in glioblastoma were detected by reverse transcription polymerase chain reaction, and expression of decoy receptor in glioblastoma was estimated. MAIN OUTCOME MEASURES： Comparison of death receptor and decoy receptor protein expression between glioblastoma and normal brain tissue; decoy receptor mRNA expression in glioblastoma. RESULTS： Death receptor protein expression was strongly positive （＋＋＋） in glioblastoma, while it was weakly positive （＋, ＋＋） in normal brain tissue. Therefore, expression rate of death receptor protein in the glioblastoma was significantly higher than that in the normal brain tissue （.~ 2 = 18.48, 23.03, P 〈 0.01）. Decoy receptor protein expression in the glioblastoma was significantly lower than that in the normal brain tissue （ x2 = 6.65, 18.76, P 〈 0.01）. The level of decoy receptor mRNA expression in glioblastoma was significantly higher than those of protein expression （ x 2 = 9.82, 10.09, P〈 0.01）. CONCLUSION： High expression of death receptor and low expression of decoy receptor are frequently observed in glioblastoma, suggesting that TRAIL receptor genes show an anti-tumor and expressive response during the initiation and development of the tumor. There are significant differences in decoy receptor expression between normal brain tissue and glioblastoma, suggesting that the restricted expression of decoy receptor in glioblastoma is regulated at the post-transcriptional level.展开更多
BACKGROUND： Under induction of retinoic acid （RA）, bone marrow stromal cells （BMSCs） can differentiate into nerve cells or neuron-like cells, which do not survive for a long time, so those are restricted to an ap...BACKGROUND： Under induction of retinoic acid （RA）, bone marrow stromal cells （BMSCs） can differentiate into nerve cells or neuron-like cells, which do not survive for a long time, so those are restricted to an application. Other neurotrophic factors can also differentiate into neuronal cells through inducing BMSCs; especially, brain-derived neurotrophic factor （BDNF） can delay natural death of neurons and play a key role in survival and growth of neurons. The combination of them is beneficial for differentiation of BMSCs. OBJECTIVE： To investigate the effects of BDNF combining with RA on inducing differentiation of BMSCs to nerve cells of adult rats and compare the results between common medium group and single BDNF group. DESIGN： Randomized controlled animal study SETTING： Department of Neurology, Affiliated Hospital of Xuzhou Medical College MATERIALS： The experiment was carried out in the Clinical Neurological Laboratory of Xuzhou Medical College from September 2003 to April 2005. A total of 24 SD rats, of either gender, 2 months old, weighing 130-150 g, were provided by Experimental Animal Center of Xuzhou Medical College [certification： SYXK （su） 2002-0038]. Materials and reagents： low-glucose DMEM medium, bovine serum, BDNF, RA, trypsin, separating medium of lymphocyte, monoclonal antibody of mouse-anti-nestin, neuro-specific enolase, glial fibrillary acidic protein （GFAP） antibody, SABC kit, and diaminobenzidine （DAB） color agent. All these mentioned above were mainly provided by SIGMA Company, GIBCO Company and Boshide Company. METHODS： Bone marrow of SD rats was selected for density gradient centrifugation. BMSCs were undertaken primary culture and subculture; and then, those cells were induced respectively in various mediums in total of 3 groups, including control group （primary culture）, BDNF group （20 μg/L BDNF） and BDNF＋RA group （20 μg/L BDNF plus 20 μg/L RA）. On the 3^rd and the 7^th days after induction, BMSCs were stained immunocytochemically with nestin （sign of nerve stem cells）, neuron-specific enolase （NSE, sign of diagnosing neurons） and GFAP （diagnosing astrocyte）, and evaluated cellular property. MAIN OUTCOME MEASURES ： Induction and differentiation in vitro of BMSCs in 3 groups RESULTS： （1） Induction and differentiation of BMSCs： Seven days after induction, cells having 2 or more apophyses were observed. Soma shaped like angle or erose form, which were similar to neurons and glial cells having strong refraction. （2） Results of immunocytochemical detection： Three days after induction, rate of positive cells in BDNF＋RA group was higher than that in BDNF group and control group [（86.15±4.58）%, （65.43±4.23）%, （4.18±1.09）%, P 〈 0.01]. Seven days after induction, rate of positive cells was lower in BDNF group and BDNF＋RA group than that in both groups at 3 days after induction [（31.12±3.18）%, （29.35±2.69）%, P 〈 0.01]; however, amounts of positive cells of NSE and GFAP were higher than those at 3 days after induction （P 〈 0.01）; meanwhile, the amount in BDNF＋RA group was remarkably higher than that in BDNF group （P 〈 0.01）. CONCLUSION： Combination of BDNF and RA can cooperate differentiation of BMSCs into neurons and astrocyte, and the effect is superior to single usage of BDNF.展开更多
Background The incidence of contrast induced nephropathy （CIN） is increasing while patients are more and more frequently undertaking coronary angiography and percutaneous coronary intervention （PCI）. CIN correlate...Background The incidence of contrast induced nephropathy （CIN） is increasing while patients are more and more frequently undertaking coronary angiography and percutaneous coronary intervention （PCI）. CIN correlates with the later cardiovascular events, the rising mortality risks and the increasing one-year target vessel revascularization. At present, few articles reported on whether the incidence of CIN induced by isotonic and hypotonic contrast agent after PCI is different or not. Objectives To investigate the different effect of isotonic and hypotonic contrast agent in inducing CIN in patients with coronary artery disease after PCI. Methods We enrolled 355 patients with coronary artery disease who undertook PCI from January 2007 to December 2008 as subjects. Renal functions of all 355 patients were normal. Patients were randomly divided into isotonic group and hypotonic group. Concentrations of serum creatinine （SCr） were measured before, 48～72 hours and 7 days （if needed） after PCI. The glomerular filtration rate （eGFR） was calculated according to MDRD formula. The incidence of CIN was defined as the concentration of SCr ≥0.5 mg/dL（44.2 μmol/L）. Hemodialysis rates and mortality were recorded in the hospital. Results There was no significant difference in the basline eGFR （79.52±5.28 vs 81.03±6.09, P0.05）, dosages of contrast agent （125.68±15.88 mL vs 123.51±16.38 mL, P0.05）, eGFR of 48-72 hours after PCI （70.26±9.48 vs 69.06±9.59, P0.05） and incidences of CIN （5.56% vs 5.78%, P0.05） between the two groups. eGFR and concentrations of SCr 7 days after PCI showed no significant difference between the two groups （P0.05）. No patient was dead or needed hemodialysis in hospital. Conclusions The effect of isotonic and hypotonic contrast agent in inducing CIN in patients with coronary artery disease after PCI has no difference.展开更多
Bone morphogenetic protein (BMP)is an efficient bone-inducing protein whichmainly exists in bone matrix.The implants of Bovine BMP (bBMP)extracted frombovine bone matrix can induce differentiation of mesenchymal cells...Bone morphogenetic protein (BMP)is an efficient bone-inducing protein whichmainly exists in bone matrix.The implants of Bovine BMP (bBMP)extracted frombovine bone matrix can induce differentiation of mesenchymal cells of mice muscle in-to chondroand osteo-cytes and form new bone further.In order to evaluate im-plantable biomaterial that possesses not only osteogenic ability,but also stengtn,展开更多
基金the Swedish Research Council(2018-02667)the National Natural Science Foundation of China(31761133015,U1704281,81901335)+3 种基金the Swedish Childhood Cancer Foundation(PR2018-0082)Swedish Governmental Grants to Scientists Working in Health Care(ALFGBG-717791)the Swedish Brain Foundation(FO2018-0034)the Chinese Scholarship Council to TL(201707040025)and to YX(201507040082)。
文摘Perinatal complications,such as asphyxia,can cause brain injuries that are often associated with subsequent neurological deficits,such as cerebral palsy or mental retardation.The mechanisms of perinatal brain injury are not fully understood,but mitochondria play a prominent role not only due to their central function in metabolism but also because many proteins with apoptosis-related functions are located in the mitochondrion.Among these proteins,apoptosis-inducing factor has already been shown to be an important factor involved in neuronal cell death upon hypoxia-ischemia,but a better understanding of the mechanisms behind these processes is required for the development of more effective treatments during the early stages of perinatal brain injury.In this review,we focus on the molecular mechanisms of hypoxic-ischemic encephalopathy,specifically on the importance of apoptosis-inducing factor.The relevance of apoptosis-inducing factor is based not only because it participates in the caspase-independent apoptotic pathway but also because it plays a crucial role in mitochondrial energetic functionality,especially with regard to the maintenance of electron transport during oxidative phosphorylation and in oxidative stress,acting as a free radical scavenger.We also discuss all the different apoptosis-inducing factor isoforms discovered,focusing especially on apoptosis-inducing factor 2,which is only expressed in the brain and the functions of which are starting now to be clarified.Finally,we summarized the interaction of apoptosis-inducing factor with several proteins that are crucial for both apoptosis-inducing factor functions(prosurvival and pro-apoptotic)and that are highly important in order to develop promising therapeutic targets for improving outcomes after perinatal brain injury.
文摘The higher frequency of varicocele in men with infertility has drawn attention and resulted in increased research at the molecular level towards treatments. The aim of this study was to investigate the role of tumor necrosis factor （TNF）-related apoptosis-inducing ligand （TRAIL） and its receptors in varicocele-induced testicular dysfunction in an experimental rat model. The rats were divided into three groups： control, sham and varicocele. Varicoceles in rats were induced by partial ligation of the left renal vein and left testes. The rats were analyzed 13 weeks after surgery. The degree of DNA fragmentation within cells in the testis was determined using terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling （TUNEL） assay. Tubule degeneration was evaluated using the Johnsen score. The expression of TRAIL and its receptors was detected by immunohistochemical and Western blotting techniques. The apoptotic index, Johnsen score and the expression of TRAIL and TRAIL receptors were examined. The data are presented as the mean-.＋s.d, and were analyzed using computer software. The KruskaI-Wallis and Dunn＇s multiple comparison tests were used in the statistical analyses. The germ cell apoptotic index was increased in rats with varicoceles when compared with the sham and control groups （P=0.0031）. The Johnsen score was significantly decreased in the varicocele group when compared with the sham and control groups （P〈O.O001）. Immunohistochemical and Western blotting analyses showed that after varicocele induction, the expression of TRAIL-R1 and TRAIL-R4 in germ cells was increased and the expression of TRAIL-R2 was decreased. There are no significant differences among the groups in terms of TRAIL and TRAIL-R3 receptor expression. The results of this study indicate that TRAIL and its receptors may have a potential role in the pathogenesis of varicocele-induced testicular dysfunction.
文摘Objective To investigate whether desferoxamine （DFO） preconditioning can induce tolerance against cerebral ischemia and its effect on the expression of hypoxia inducible factor 1 α （HIF- 1α） and erythropoietin （EPO） in vivo and in vitro. Methods Rat model of cerebral ischemia was established by middle cerebral artery occlusion with or without DFO administration. Infarct size was examined by TTC staining, and the neurological severity score was evaluated according to published method. Cortical neurons were cultured under ischemia stress which was mimicked by oxygen-glucose deprivation （OGD）, and the neuron damage was assessed by MTT assay. Immunofluorescent staining was employed to detect the expressions of HIF-1 and EPO. Results The protective effect induced by DFO （decreasing the infarction volume and ameliorating the neurological function） appeared at 2 d after administration ofDFO （post-DFO）, lasted until 7 d and disappeared at 14 d （P 〈 0.05）; the most effective action was observed at 3 d post-DFO. DFO induced tolerance of cultured neurons against OGD： neuronal viability was increased 23%, 34%, 40%, 48% and 56% at 8 h, 12 h, 24 h, 36 h, and 48 h, respectively, post-DFO （P 〈 0.05）. Immunofluorescent staining found that HIF-1 α and EPO were upregulated in the neurons of rat brain at 3 d and 7 d post-DFO; increase of HIF-1 α and EPO appeared in cultured cortex neurons at 36 h and 48 h post-DFO. Conclusion DFO induced tolerance against focal cerebral ischemia in rats, and exerted protective effect on OGD cultured cortical neurons. DFO significant induced the expression of HIF- 1 α and EPO both in vivo and in vitro. DFO preconditioning can protect against cerebral ischemia, which may be associated with the synthesis of HIF- 1 α and EPO.
文摘AIM:To study the effects of Helicobacter pylori(H. pylori)tumor necrosis factor-α(TNF)inducing protein (Tip-α)on cytokine expression and its mechanism. METHODS:We cloned Tip-αfrom the H.pylori strain 26695,transformed Escherichia coli with an expression plasmid,and then confirmed the expression product by Western blotting.Using different concentrations of Tip-αthat affected SGC7901 and GES-1 cells at different times,we assessed cytokine levels using enzyme-linked immunosorbent assay.We blocked SGC7901 cells with pyrrolidine dithiocarbamate(PDTC),a specific inhibitor of nuclear factorκB(NF-κB).We then detected interleukin(IL)-1βand TNF-αlevels in SGC7901 cells. RESULTS:Western blot analysis using an anti-Tip-α antibody revealed a 23-kDa protein,which indicated that recombinant Tip-αprotein was recombined successfully.The levels of IL-1β,IL-8 and TNF-αwere sig-nificantly higher following Tip-αinterference,whether GES-1 cells or SGC-7901 cells were used(P<0.05).However,the levels of cytokines(including IL-1β,IL-8 and TNF-α)secreted by SGC-7901 cells were greater than those secreted by GES-1 cells following treatment with Tip-αat the same concentration and for the same duration(P<0.05).After blocking NF-κB with PDTC, the cells(GES-1 cells and SGC-7901 cells)underwent interference with Tip-α.We found that IL-1βand TNF-αlevels were significantly decreased compared to cells that only underwent Tip-αinterference(P<0.05). CONCLUSION:Tip-αplays an important role in cyto-kine expression through NF-κB.
基金Supported by Natural Science Foundation of Fujian Province(Based on NLRP3 Inflammatory Body/Caspase-1-mediated Gastric Epithelial Cell Death to Explore the Molecular Mechanism of Qinghua Decoction in the Treatment of Chronic Atrophic Gastritis,No.2020J01253)。
文摘OBJECTIVE:To examine the efficacy of Qinghuayin(清化饮,QHY)in rat chronic atrophic gastritis(CAG)models and explored the molecular mechanism of QHY in treating CAG.METHODS:In total,65 Wistar rats were randomly divided into the control(n=10)and CAG groups(n=55).CAG model rats were further divided into five groups:model(n=10),vitacoenzyme(n=10),low-dose QHY(n=10),medium-dose QHY(n=10),and high-dose QHY groups(n=10).We analyzed histopathological changes using hematoxylin and eosin staining and measured interleukin(IL)-6 and IL-8 levels in serum using enzyme-linked immunosorbent assay(ELISA)(Boster Bio,Pleasanton,USA).In addition,gastrin(GAS),pepsinogen I(PGI),and PGII expressions were evaluated using ELISA.The protein and m RNA expression of toll-like receptor 4(TLR4)and toll or interleukin-1 receptor domaincontaining adaptor inducing interferon-β(TRIF)was detected by Western blotting and quantitative reverse transcription-polymerase chain reaction,respectively.RESULTS:Our results revealed that histopathological changes in CAG model rates could be restored by low-,medium-,and high-dose QHY.The changes in GAS and PGI/II expression demonstrated that QHY improved CAG.Serum IL-6 and IL-levels were decreased by QHY administration.TLR4 and TRIF were upregulated at the m RNA and protein levels in the model group but downregulated by QHY administration.CONCLUSION:We concluded that QHY could effectively improve the histopathological changes of the gastric mucosa induced by CAG in rats.The therapeutic mechanism of QHY may be related to inhibition of the inflammatory factors IL-6 and IL-8 and suppression of TLR4/TRIF m RNA and protein expression.
基金supported by the project of National Junior Research Fellowship,which National Research Foundation of Korea conducts from 2012
文摘Acupuncture has been broadly used as a major therapeutic tool for at least 2,100 years in East Asia. Acupuncture is used for improving the pathological condition of remote organs, such as stomach, liver, brain, etc. by stimulating skin and muscle with fine metal needles through meridians which have been regarded as pathways of Qi within the body （Ernst, 2006）. Physicians in East Asia have tried to treat many kinds of diseases by acupuncture and it was considerably success- ful. Various diseases for which acupuncture is known to be effective, are neurological diseases, including sequelae of stroke, Alzheimer＇s disease （AD）, Parkinson＇s disease （PD）, and so on （Nam et al., 2013）.
基金supported by the National Natural Science Foundation of China(Nos.81974151,31972908,81991500,and 81991502)。
文摘Immunoglobulin G4-related sialadenitis(IgG4-RS)is an immune-mediated fibro-inflammatory disease and the pathogenesis is still not fully understood.The aim of this study was to explore the role and mechanism of interleukin-13(IL-13)in the cellular senescence during the progress of IgG4-RS.We found that the expression of IL-13 and IL-13 receptorα1(IL-13Rα1)as well as the number of senescent cells were significantly higher in the submandibular glands(SMGs)of IgG4-RS patients.IL-13 directly induced senescence as shown by the elevated activity of senescence-associatedβ-galactosidase(SA-β-gal),the decreased cell proliferation,and the upregulation of senescence markers(p53 and p16)and senescence-associated secretory phenotype(SASP)factors(IL-1βand IL-6)in SMG-C6 cells.Mechanistically,IL-13 increased the level of phosphorylated signal transducer and activator of transcription 6(p-STAT6)and mitochondrial-reactive oxygen species(mt ROS),while decreased the mitochondrial membrane potential,ATP level,and the expression and activity of superoxide dismutase 2(SOD2).Notably,the IL-13-induced cellular senescence and mitochondrial dysfunction could be inhibited by pretreatment with either STAT6 inhibitor AS1517499 or mitochondria-targeted ROS scavenger Mito TEMPO.Moreover,IL-13 increased the interaction between p-STAT6 and c AMP-response element binding protein(CREB)-binding protein(CBP)and decreased the transcriptional activity of CREB on SOD2.Taken together,our findings revealed a critical role of IL-13 in the induction of salivary gland epithelial cell senescence through the elevated mitochondrial oxidative stress in a STAT6–CREB–SOD2-dependent pathway in IgG4-RS.
基金Supported by National Natural Science Foundation of China:Mechanism of Protective Effects of Moxibustion on Regulating m TOR Signaling Pathway and Inhibiting Autophagy in Rats(No.81574084)Key Research and Development Program of Anhui Province:Mechanism and Clinical Application of Moxibustion in that Treatment of Chronic Heart Failure(No.202004j07020045)+1 种基金Open Fund Project of Key Laboratory of Xin’an Medical Education Ministry:Effect of Moxibustion on Myocardial Cell Energy Metabolism in Chronic Heart Failure Patients Based on Xin’an Physician’s Theory of Strengthening the Foundation and Cultivating the Original(No.2020xayx07)Key Natural Science Projects of Anhui Provincial Education Department:Mechanism of Moxibustion Against CHF Fibrosis Based on mi R-21/PTEN/m TOR Signaling Pathway-mediated Regulation of Autophagy in Myocardial Cells by circ PAN3(No.KJ2021A0570)。
文摘OBJECTIVE:To observe the effects of moxibustion at bilateral Feishu(BL13)and Xinshu(BL15)combined with benazepril on myocardial cells apoptosis index,the expression levels of apoptosis-related proteins cytochrome c(Cyt-C)and apoptosis-inducing factor(AIF)in chronic heart failure(CHF)rats.METHODS:Sixty-five rats were randomly divided into normal group(n=10)and model-I group(n=55).After modeling,CHF rats in model-I group were divided into model group,moxibustion group,benazepril group,moxibustion plus benazepril group(abbreviated as aibei group,the same below),10 rats in each group.Echocardiogram index was examined by echocardiography.Hemodynamic indices were measured by rat cardiac function meter.Serum B-type brain natriuretic peptide(BNP)was detected by enzymelinked immunosorbent assay.Myocardial cells apoptosis index was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling staining.Pathological changes of myocardial tissues were observed by hematoxylin and eosin staining.The expression levels of Cyt-C and AIF in myocardial tissues were detected by Western blot.RESULTS:Compared with normal group,ejection fraction and left ventricular diameter shortening rate in model-Ⅰgroup were significantly reduced,myocardial cells of rats in model group exhibited unclear transverse striations,cells swellings and vacuoles,cardiac functions were deteriorated,serum BNP level,myocardial cells apoptosis index,and the expression levels of Cyt-C and AIF were significantly increased.Compared with model group,myocardial cells of rats in moxibustion group,benazepril group,and aibei group were dyed more evenly,muscle fibers were arranged relatively neatly,cardiac functions were improved,serum BNP level,myocardial cells apoptosis index,and the expression levels of Cyt-C and AIF were significantly decreased.Compared with aibei group,cardiac functions were worsened,myocardial cells apoptosis index,and the expression levels of Cyt-C and AIF were increased.CONCLUSION:Moxibustion at bilateral Feishu(BL13)and Xinshu(BL15)combined with benazepril could improve CHF better than moxibustion at bilateral Feishu(BL13)and Xinshu(BL15)or benazepril alone.The mechanisms might be that they can inhibit the expressions of Cyt-C and AIF,and inhibit the apoptosis of cardiomyocytes.
文摘The corneal crosslinking （CXL） with riboflavin and ultraviolet-A （UVA） is a new therapy method to successfully treat infectious keratitis in clinical practice. However, there are rare reports on the complications of CXL such as the secondary keratitis. The diverse clinical outcomes on keratitis have highlighted the necessity to further evaluate the efficacy and complications of CXL. We reviewed the positive and negative reports on UVA/riboflavin related with keratitis and provided our opinion on the therapeutic and side effect of UVA/riboflavin crosslinking on keratitis.
文摘Programmed cell death （PCD） signaling pathways are import- ant contributors to acute neurological insults such as hypox- ic-ischemic brain damage, traumatic brain injury, stroke etc. The pathogenesis of all these diseases is closely linked with ab- erration of apoptotic cell death pathways. Mitochondria play a crucial role during PCD, acting as both sensors of death signals, and as initiators of biochemical path- ways, which cause cell death （Bras et al., 2005）. Cytochrome c was the firstly identified apoptogenic factor released from mitochondria into the cytosol, where it induces apoptosome formation through the activation of caspases. Other proteins, such as apoptosis inducing factor （AIF）, have been subsequently identified as mitochondrial released factors. AIF contributes to apoptotic nuclear DNA damage （Bras et al., 2005）. in a caspase-independent way
文摘It was found that the growth of malignant tumour in mice was inhibited and the ability of immune cell’s dissolving cancer cells was enhanced by ultralow frequency (ULF) pulsed gradient magnetic field. The DNA contents of nuclei decreased which indicated that magnetic field can block DNA replication and mitosis of cancer cells. It was observed that magnetic field inhibited the cancer cell’s metabolism, lowered its malignancy, and restrained its rapid and heteromorphic growth. The morphology properties of Programmed Cell Death (PCD) of the cancer cells of the treated group by magnetic field was observed for the first time. The heterochromatin condensed and coagulated together along the nuclear membrane; the endoplasmic reticulums expanded and fused with the cellular membrane; many apoptotic bodies which were packed by the cellular membrane appeared and were devoured by the lymphocytes and plasma.
文摘Since the welded constructions produce easily stress corrosion cracking (SCC) or fatigue disruption in corrosive medium or under ripple load, two methods inducing compressive stress on structural surfaces by anti-welding-heating treatment (AWHT) and explosion treatment (ET) are presented. The results show that they are good ways to resisting SCC on the welded SS400 steel or other components.
文摘During our 4-year work in The Republic of Yemen, the authors used acupuncture therapy to restore consciousness and induce resuscitation plus scalp acupuncture in 80 cases of acute apoplexy, with 63 cases cured, accounting for 78.75%. This therapy is obviously superior to the traditional principle of point selection in acupuncture used in 25 cases as to curative effect, treatment course and other aspects. There is an obvious statistical difference (P【 0.005).
基金Supported by the National Natural Science Foundation of China under Grant No 61875235the Ph.D. Programs Foundation of Ministry of Education of China under Grant No 20130009110008the Beijing Municipal Education Commission Project under Grant No KM201210015008
文摘The effect of formed CH_3NH_3 at the heterojunction on properties of CH_3NH_3PbI_3 material is investigated based on experiment and theoretical calculation. Our calculation results show that the giant dielectric constant, anomalous hysteresis and long-lasting polarization for CH_3NH_3PbI_3 originate from the formed CH_3NH_3 at the heterojunction. It is found that the induced weak EPS by the reorientation of CH_3NH_3 sub-group along the built-in electric field enables us to effectively increase the ordering of entire lead-halide framework. In addition, the heterojunction has an advantage of channel separation between carrier transport and electron diffusion. These properties of the heterojunction are the main origin of the high efficiency of CH_3NH_3PbI_3 solar cells.
基金the National Natural Science Foundation of China, No. 30772537
文摘BACKGROUND： Previous studies have reported that statins are less toxic to the human body and have greater antitumor activity; however, few studies have addressed the antitumor effect of statins combined with tumor necrosis factor-related apoptosis inducing ligand （TRAIL）. OBJECTIVE： To explore the effect of TRAIL combined with mevastatin on the proliferation and apoptotic cell death of a human glioma cell line SWO-38, and to study its mechanism of action. DESIGN, TIME AND SETTING： An in vitro control experiment was performed at the Central Laboratory of the Third Hospital Affiliated to Sun Yat-sen University, between January and April 2009. MATERIALS： The human SWO-38 cell line was provided by Cell Research, Department of Animal Experimental Center of Sun Yat-sen University; human recombinant soluble TRAIL by R＆D, USA; and mevastatin by Sigma, USA. METHODS： SWO-38 cells were separately incubated in TRAIL （100, 200, 300, 400, and 500 tJg/L） and mevastatin （5, 10, 20, 30, and 40 pmol/L） for 72 hours. In addition, SWO-38 cells were incubated in TRAIL （300 μg/L）, mevastatin （30 μmol/L）, and a solution containing both TRAIL and mevastatin for 12, 24, 48 and 72 hours. MAIN OUTCOME MEASURES： Cell proliferation was detected using methyl thiazolyl tetrazolium assay; cell apoptosis was observed using Hoechst 33258 staining and fluorescence microscopy and was measured using Annexin V/propidium iodide flow cytometry; TRAIL R1/DR4 and TRAIL R2/DR5 protein expressions levels were measured using indirect immunofluorescence staining combined with flow cytometry in the recombinant soluble TRAIL （rsTRAIL, 300 tJg/L）, mevastatin （30 IJmol/L） and combination groups; TRAIL R1/DR4 and TRAIL R2/DR5 mRNA expression was detected using real-time polymerase chain reaction. RESULTS： rsTRAIL, mevastatin and their combination inhibited tumor proliferation in a time- and dose-dependent manner. The proliferation inhibitory rate and apoptosis rate of human SWO-38 cells in the combined group were significantly greater than the rsTRAIL or mevastatin alone group （P 〈 0.01）. TRAIL R1/DR4 and TRAIL R2/DR5 protein and mRNA expressions were increased in the combination group compared with mevastatin or rsTRAIL alone after 72 hours （P 〈 0.01）. CONCLUSION： Both rsTRAIL and mevastatin inhibit the proliferation and apoptosis of the human glioma cell line SWO-38, while their combination enhances the anti-tumor effect. The mechanism of action possibly correlates to the upregulation of TRAIL R1/DR4 and TRAIL R2/DR5 mRNA expression by mevastatin, thereby enhancing the cell sensitivity to rsTRAIL.
基金Key Program of Tenth Five-Year Plan and the 211 Key Subject Construction Foundation, No. 2002-2
文摘BACKGROUND： Receptors for tumor necrosis factor related apoptosis inducing ligand （TRAIL） include death receptor 4, death receptor 5, decoy receptor 1, and decoy receptor 2. Activation of death receptor 4 and 5 selectively kills tumor cells. OBJECTIVE： To detect TRAIL receptor expression in glioblastoma by immunohistochemistry and RT-PCR, and to compare this expression to that in normal brain tissue. DESIGN： Observational analysis. SETTING： Department of Pathology, the First Affiliated Hospital of Zhengzhou University; Henan Tumor Pathology Key Laboratory. PARTICIPANTS： Twenty-five patients （17 males and 8 females） who received glioblastoma resection were selected from the Fifth Affiliated Hospital of Zhengzhou University, between September 2003 to June 2004. All glioblastoma samples were diagnosed pathologically. Twenty patients （12 males and 8 females） with craniocerebral injury who received normal brain tissue resection were selected in the same time period. There were no significant differences in sex and age between glioblastoma patients or between craniocerebral injury patients （P 〉 0.05）. All patients and appropriate relatives provided informed consent, and this study was approved by the local research ethics committee. METHODS： Polyclonal antibody against TRAIL receptors and an immunohistochemical kit （batch number： 200502） were purchased from Boster Company, Wuhan. Immunohistochemistry： Expression of death receptor 4, death receptor 5, decoy receptor l, and decoy receptor 2 were observed in both glioblastoma and normal brain tissue. The experiment was performed according to the kit instructions, and positive staining was brown-yellow. Assessment： There were no positive signals （-）; weakly positive signals, positive cells 〈 25% （＋）; weakly positive signals, positive cells 25%-50% （＋＋）; strongly positive signals, positive cells 50%-75% （＋＋＋）; strongly positive signals, positive cells 〉 75% （＋＋＋＋）. Evaluation： Expression levels of TRAIL receptors were estimated in both normal brain tissue and glioblastoma. Expression of decoy receptor 1 and decoy receptor 2 mRNA in glioblastoma were detected by reverse transcription polymerase chain reaction, and expression of decoy receptor in glioblastoma was estimated. MAIN OUTCOME MEASURES： Comparison of death receptor and decoy receptor protein expression between glioblastoma and normal brain tissue; decoy receptor mRNA expression in glioblastoma. RESULTS： Death receptor protein expression was strongly positive （＋＋＋） in glioblastoma, while it was weakly positive （＋, ＋＋） in normal brain tissue. Therefore, expression rate of death receptor protein in the glioblastoma was significantly higher than that in the normal brain tissue （.~ 2 = 18.48, 23.03, P 〈 0.01）. Decoy receptor protein expression in the glioblastoma was significantly lower than that in the normal brain tissue （ x2 = 6.65, 18.76, P 〈 0.01）. The level of decoy receptor mRNA expression in glioblastoma was significantly higher than those of protein expression （ x 2 = 9.82, 10.09, P〈 0.01）. CONCLUSION： High expression of death receptor and low expression of decoy receptor are frequently observed in glioblastoma, suggesting that TRAIL receptor genes show an anti-tumor and expressive response during the initiation and development of the tumor. There are significant differences in decoy receptor expression between normal brain tissue and glioblastoma, suggesting that the restricted expression of decoy receptor in glioblastoma is regulated at the post-transcriptional level.
文摘BACKGROUND： Under induction of retinoic acid （RA）, bone marrow stromal cells （BMSCs） can differentiate into nerve cells or neuron-like cells, which do not survive for a long time, so those are restricted to an application. Other neurotrophic factors can also differentiate into neuronal cells through inducing BMSCs; especially, brain-derived neurotrophic factor （BDNF） can delay natural death of neurons and play a key role in survival and growth of neurons. The combination of them is beneficial for differentiation of BMSCs. OBJECTIVE： To investigate the effects of BDNF combining with RA on inducing differentiation of BMSCs to nerve cells of adult rats and compare the results between common medium group and single BDNF group. DESIGN： Randomized controlled animal study SETTING： Department of Neurology, Affiliated Hospital of Xuzhou Medical College MATERIALS： The experiment was carried out in the Clinical Neurological Laboratory of Xuzhou Medical College from September 2003 to April 2005. A total of 24 SD rats, of either gender, 2 months old, weighing 130-150 g, were provided by Experimental Animal Center of Xuzhou Medical College [certification： SYXK （su） 2002-0038]. Materials and reagents： low-glucose DMEM medium, bovine serum, BDNF, RA, trypsin, separating medium of lymphocyte, monoclonal antibody of mouse-anti-nestin, neuro-specific enolase, glial fibrillary acidic protein （GFAP） antibody, SABC kit, and diaminobenzidine （DAB） color agent. All these mentioned above were mainly provided by SIGMA Company, GIBCO Company and Boshide Company. METHODS： Bone marrow of SD rats was selected for density gradient centrifugation. BMSCs were undertaken primary culture and subculture; and then, those cells were induced respectively in various mediums in total of 3 groups, including control group （primary culture）, BDNF group （20 μg/L BDNF） and BDNF＋RA group （20 μg/L BDNF plus 20 μg/L RA）. On the 3^rd and the 7^th days after induction, BMSCs were stained immunocytochemically with nestin （sign of nerve stem cells）, neuron-specific enolase （NSE, sign of diagnosing neurons） and GFAP （diagnosing astrocyte）, and evaluated cellular property. MAIN OUTCOME MEASURES ： Induction and differentiation in vitro of BMSCs in 3 groups RESULTS： （1） Induction and differentiation of BMSCs： Seven days after induction, cells having 2 or more apophyses were observed. Soma shaped like angle or erose form, which were similar to neurons and glial cells having strong refraction. （2） Results of immunocytochemical detection： Three days after induction, rate of positive cells in BDNF＋RA group was higher than that in BDNF group and control group [（86.15±4.58）%, （65.43±4.23）%, （4.18±1.09）%, P 〈 0.01]. Seven days after induction, rate of positive cells was lower in BDNF group and BDNF＋RA group than that in both groups at 3 days after induction [（31.12±3.18）%, （29.35±2.69）%, P 〈 0.01]; however, amounts of positive cells of NSE and GFAP were higher than those at 3 days after induction （P 〈 0.01）; meanwhile, the amount in BDNF＋RA group was remarkably higher than that in BDNF group （P 〈 0.01）. CONCLUSION： Combination of BDNF and RA can cooperate differentiation of BMSCs into neurons and astrocyte, and the effect is superior to single usage of BDNF.
文摘Background The incidence of contrast induced nephropathy （CIN） is increasing while patients are more and more frequently undertaking coronary angiography and percutaneous coronary intervention （PCI）. CIN correlates with the later cardiovascular events, the rising mortality risks and the increasing one-year target vessel revascularization. At present, few articles reported on whether the incidence of CIN induced by isotonic and hypotonic contrast agent after PCI is different or not. Objectives To investigate the different effect of isotonic and hypotonic contrast agent in inducing CIN in patients with coronary artery disease after PCI. Methods We enrolled 355 patients with coronary artery disease who undertook PCI from January 2007 to December 2008 as subjects. Renal functions of all 355 patients were normal. Patients were randomly divided into isotonic group and hypotonic group. Concentrations of serum creatinine （SCr） were measured before, 48～72 hours and 7 days （if needed） after PCI. The glomerular filtration rate （eGFR） was calculated according to MDRD formula. The incidence of CIN was defined as the concentration of SCr ≥0.5 mg/dL（44.2 μmol/L）. Hemodialysis rates and mortality were recorded in the hospital. Results There was no significant difference in the basline eGFR （79.52±5.28 vs 81.03±6.09, P0.05）, dosages of contrast agent （125.68±15.88 mL vs 123.51±16.38 mL, P0.05）, eGFR of 48-72 hours after PCI （70.26±9.48 vs 69.06±9.59, P0.05） and incidences of CIN （5.56% vs 5.78%, P0.05） between the two groups. eGFR and concentrations of SCr 7 days after PCI showed no significant difference between the two groups （P0.05）. No patient was dead or needed hemodialysis in hospital. Conclusions The effect of isotonic and hypotonic contrast agent in inducing CIN in patients with coronary artery disease after PCI has no difference.
文摘Bone morphogenetic protein (BMP)is an efficient bone-inducing protein whichmainly exists in bone matrix.The implants of Bovine BMP (bBMP)extracted frombovine bone matrix can induce differentiation of mesenchymal cells of mice muscle in-to chondroand osteo-cytes and form new bone further.In order to evaluate im-plantable biomaterial that possesses not only osteogenic ability,but also stengtn,
