Investigating the expression profiles of cysteine string proteins(CSPs)in cochlear tissue

Objective:This study aims to explore the expression patterns of cysteine string protein alpha(CSPα)and cysteine string protein beta(CSPβ)in the mammalian inner ear,with an emphasis on their temporal dynamics during the developmental stages of C57BL/6 mice.Methods:We utilized immunofluorescence staining to assess the localization and distribution of CSPαand CSPβwithin the inner ears of C57BL/6 mice and miniature pigs.Additionally,this method facilitated the investigation of their temporal expression profiles.Results:In adult C57BL/6 mice and miniature pigs,CSPαand CSPβwere identified in the cytoplasm of inner hair cells and spiral ganglion cells,yet were absent in outer hair cells.Both proteins were found to colocalize with Ctbp2 on the basal side of the cytoplasm in inner hair cells’basilar membrane.Expression of CSPαwas observed at the nerve fiber termini at the basilar membrane’s base of inner and outer hair cells 10 days postnatally in C57BL/6 mice.Notably,expression of both CSPαand CSPβin the cytoplasm of inner hair cells emerged on the 12th day post-birth,aligning with the timeline for registering cochlear potentials.The expression levels of both proteins increased with age,but were consistently absent in outer hair cells.Contrastingly,expression of CSPαand CSPβwas present in the cytoplasm of inner hair cells in miniature pigs as early as one day post-birth,yet remained absent in the three rows of outer hair cells.Conclusion:CSPαand CSPβexhibit predominant and specific expression in inner hair cells and spiral ganglion cells.A unique expression pattern was observed for CSPα,which was also present at the nerve fiber endings of both inner and outer hair cells.The developmental expression trajectory of CSPαand CSPβin mouse inner hair cells is characterized by an initial absence,followed by a gradual increase.Moreover,the timing of expression onset between mice and miniature pigs indicates distinct temporal dynamics,suggesting a potential role in auditory development.

Tumor-related factor complement Clq/TNF-related protein 6 affects the development of digestive system tumors through the phosphatidylinositol 3-kinase pathway

In this editorial,we review the work of Razali et al published in World J Gas-troenterology,with a particular focus on the effect of rs10889677 variation in the phosphatidylinositol 3-kinase(PI3K)pathway and buparlisib on colitis-associated cancer.The role of PI3K in promoting cancer progression has been widely recognized,as it is involved in regulating the survival,differentiation,and prolif-eration of cancer cells.The complement Clq/TNF-related protein 6(CTRP6)is a newer tumor-associated factor.Recent studies have revealed the pro-tumor effect of CTRP6 in gastric cancer,hepatocellular carcinoma,colorectal cancer,and other gastrointestinal tumors through the PI3K pathway.This article attempts to reveal the mechanism through which the CTRP6 affects the development of digestive system tumors through the PI3K pathway by summarizing recent research.

Effects of realgar on stress proteins, inflammatory mediators, and complement in brain tissue and serum of rats with inflammatory brain injury

BACKGROUND：The Chinese herbal compound realgar exerts detoxification effects as an adjuvant. It is suggested that realgar exerts detoxification via the following pathways： in the pathological state, realgar corrects the oxidative stress state by increasing stress levels, activating some endogenous protective factors and antagonizing the excessive release of inflammatory factors, as well as inhibiting complement activation. OBJECTIVE： To observe the changes in stress proteins, inflammatory mediators, and complement in the brain tissue and serum of rats with inflammatory brain injury, which have been treated with the Chinese herbal compound Angong Niuhuang, and to compare the efficacy of Angong Niuhuang with that of realgar, to verify the mechanism of action of realgar. DESIGN, TIME AND SETTING： Randomized, controlled, cytological experiment, performed in the Institute of Clinical Pharmacology, Guangzhou University of Traditional Chinese Medicine in March 2006. MATERIALS： Thirty-six healthy, male, Sprague Dawley rats received 250 U/kg Bordetella pertussis via the common carotid artery within 15 seconds to induce inflammatory brain injury. Reagents and kits were as follows： Realgar and Angong Niuhuang powder （Foshan Second Pharmaceutical Factory, China）, Bordetella pertussis diagnostic antigen （National Institute for the Control of Pharmaceutical and Biological Products, China）, heat shock protein 70 （HSP70） enzyme-labeled immunosorbent assay （ELISA） kit （Stressgen, USA）, tumor necrosis factor-α （TNF-α） ELISA kit （Biosource, USA）, nitric oxide synthase （NOS） kit, Coomassie brilliant blue protein kit （Nanjing Jiancheng Bioengineering Co.,Ltd., China）, and complements C3 and C4 （Shanghai Kehua Dongling Diagnositic Products Co.,Ltd., China）. METHODS： Thirty-six rats were randomly and evenly divided into the following six groups： normal control, model, high-, middle-, and low-dose realgar-treated, and Angong Niuhuang-treated groups. At one hour prior to establishing the model, rats in the high-, middle-, and low-dose realgar-treated groups were administered 300, 150, and 75 mg/kg realgar, respectively; while rats in the Angong Niuhuang group received 1.5 g/kg Angong Niuhuang powder; In the model group, rats were administered Bordetalla pertussis only. MAIN OUTCOME MEASURES： Two hours after the administration of Bordetalla pertussis, the HSP70 level in the brain tissue and serum levels of interleukin-1β （IL-1β）, interleukin-6 （IL-6）, and TNF-α were determined by ELISA tests, hemooxygenase-1 （HO-1） activity in the brain tissue and serum was determined by dual-wavelength spectrophotometry, NOS and inducible nitric oxide synthase （iNOS） activities in the brain tissue were measured by the Bradford assay method, and serum levels of complement C3 and C4 were determined by immunoturbidimetry. RESULTS： In the high-dose realgar and Angong Niuhuang groups, the HSP70 level in the brain tissue of rats with inflammatory brain injury was increased significantly （P 〈 0.01）. In the realgar-treated groups, HO-1 activity in the brain tissue and serum was dose-dependently enhanced with increasing realgar doses. However, no significant difference existed between the realgar groups and the model group （P 〉 0.05）. In the Angong Niuhuang group, HO-1 activity in the brain tissue and serum was increased （P 〈 0.05）. In the realgar and Angong Niuhuang groups, serum levels of IL-1β, IL-6, and TNF-α were significantly decreased; the serum IL-1β level recovered to the normal level and serum levels of IL-6 and TNF-α dose-dependently decreased in the realgar groups. NOS activity in the brain tissue was lower in the high-dose realgar group than in the model group （P 〈 0.05）. iNOS activity was significantly lower in the middle- and high-dose realgar groups than in the model group （P 〈 0.01）. NOS and iNOS activities were significantly lower in the Angong Niuhuang group than in the model group （P 〈 0.01）. The serum C3 level was significantly decreased in the middle-dose realgar and Angong Niuhuang groups （P 〈 0.01）. CONCLUSION： At certain doses, realgar is able to correct the oxidative stress state, by inducing and activating endogenous protective factors, such as HSP70, and inhibiting the excessive release of inflammatory mediators, such as IL-1β, IL-6, and TNF-α. However, it remains unclear whether realgarinhibits the activation of C3 and C4.

The Synergistic Effect of HT and Complement Regulatory Proteins in Resisting the Immunological Rejection of Heterogenic Transplantation

Objective：To establish the polytransgenic mice expressing the human HT and complement regulatory proteins （CRPs） and discuss their ability to resist the hyperacute rejection （HAR） and delayed xenograft rejection （DXR） of heterogenic transplantation. Methods ：Transgenic mice were produced by microinjection to construct gene for human HT, delay acceleration factor （DAF） and/or CD59 into the male pronucleus of zygote. PCR and Southern blot were used to screen the positive trarisgenic mice. Flow cytometry （FCM） was used to detect the expression of HT, ct-Gal and DAF or CD59 on the PBMCs of transgenic mice. The survival time and function of the heart of transgenic mice were determined by a modified Langendorff cardiac perfusion apparatus： The change of proteinosis on IgM,IgG, C3c and C9 from different cardiac vascular iendothelial cells of transgenic mice were detected by immunohistochemistry. Results：HT, DAF or CD59 were highly expressed on the positive transgenic mice by FCM. The deposition of IgM,IgG,C3c or C9 in the cardiac vascular endothelial cells of the positive transgenic mice were de- creased. The survival time and function of the heart of the co-transgenic mice with AB serum perfusion were significantly longer and higher than that of the single HT positive transgenic mice（P 〈0.05）. Conclusion ：The mice co-expressing HT/DAF or HT/CD59 could resist the HAR,which was better than those expressing HT alone. It is feasible to use HT and CRPs co-transgenic methods to resist the HAR and DXR.

Myocardial Infarction and Intracerebral Hemorrhage in a Chinese Population: Relationship with Lipoproteins and Adipokines

BACKGROUND: Adipokines and inflammatory factors play an important role in disease progression. Two cardiovascular diseases which have important contributions to mortality and morbidity in China are in-tracerebral hemorrhage (ICH) and myocardial infarction (MI). Acylation stimulating protein has been shown in North American populations to have strong associations with risk factors for MI. Complement C3 (C3) a component of the innate complement immune system is the precursor protein to ASP;C3 has been impli-cated in the pathogenesis of ICH. OBJECTIVE: In this case-control study we examined the association be-tween BMI, lipoproteins adiponectin, C3 and ASP) in a Chinese population. METHODS AND RESULTS: Three groups of subjects were studied: ICH group (N = 41), MI group (N = 60) and a control group (N = 44). There was no difference in BMI for either ICH or MI compared to controls (Control: 22.3 ± 0.3 kg/m2;ICH: 21.3 ± 0.4 vs MI: 22.5 ± 0.2, ICH and MI versus control pNS). The ICH group had lower LDL-C (Control: 3.21 ± 0.13 mmol/L;ICH: 2.54 ± 0.13;MI: 2.99 ± 0.13;ICH vs control p < 0.05), total cholesterol (Control: 5.06 ± 0.16 mmol/L;ICH: 4.40 ± 0.15;MI: 4.51 ± 0.14;ICH and MI vs control p < 0.05),, HDL-C (Control: 1.34 ± 0.05 mmol/L;ICH: 1.22 ± 0.06;MI: 0.95 ± 0.04;ICH and MI vs control p < 0.05), and C3 (Control: 2.58 ? 0.21 g/L;ICH: 1.85 ? 0.19;MI: 2.87 ? 0.16;ICH vs control p < 0.05), and higher TG (Control: 1.10 ± 0.07 mmol/L;ICH: 1.77 ± 0.17;MI: 1.61 ± 0.10, ICH and MI vs control p < 0.05), compared to the controls. The MI group had lower total cholesterol and HDL-C and higher TG and ASP (Control: 33.70 ? 2.07 nM;ICH: 35.10 ? 2.33;MI: 41.50 ? 1.81;MI vs control p < 0.05) compared to control. CONCLUSION: Chinese men and women who had an MI displayed elevated ASP unrelated to an increase in the precursor protein, C3. Chinese men and women with ICH had ASP levels similar to controls yet lower C3 suggesting that C3, and the regulation of C3 conversion to ASP may be important in ICH disease pathology.

Association of C-reactive protein and complement factor H gene polymorphisms with risk of lupus nephritis in Chinese population

BACKGROUND Complement overactivation is a major driver of lupus nephritis(LN).Impaired interactions of C-reactive protein(CRP)with complement factor H(CFH)have been shown as a pathogenic mechanism that contributes to the overactivation of complement in LN.However,genetic variations of neither CRP nor CFH show consistent influences on the risk of LN.AIM To examine whether genetic variations of CRP and CFH in combination can improve the risk stratification in Chinese population.METHODS We genotyped six CRP single nucleotide polymorphisms(SNPs)(rs1205,rs3093062,rs2794521,rs1800947,rs3093077,and rs1130864)and three CFH SNPs(rs482934,rs1061170,and rs1061147)in 270 LN patients and 303 healthy subjects.RESULTS No linkage was found among CRP and CFH SNPs,indicating lack of genetic interactions between the two genes.Moreover,CRP and CFH SNPs,neither individually nor in combination,are associated with the risk or clinical manifestations of LN.Given the unambiguous pathogenic roles of the two genes.CONCLUSION These findings suggest that the biological effects of most genetic variations of CRP and CFH on their expressions or activities are not sufficient to influence the disease course of LN.

Phosphorylation and biosynthesis of high molecular weight proteins of tumor nuclear matrix

Our previous studies showed a predominance of high molecular weight protein group in tumor nuclear matrices. Contrary to normal cells, proteins of this group are preferentially phosphorylated. Phosphoproteins of hepatoma nuclear matrix are selectively subjected to rapid proteolysis. By alkali treatment and a monoclonal antibody against phosphotyrosyl residue the presence of two high molecular weight bands of phosphotyrosyl-containing proteins was detected in nuclear matrices of tumor but not of normal liver cells. High molecular weight protein group of tumor nuclear matrices revealed also a rapid turnover and preferential incorporation of labeled amino acids selectively inhibited by chloramphenicol.

Tea polyphenols increase X-ray repair cross-complementing protein 1 and apurinic/apyrimidinic endonuclease/redox factor-1 expression in the hippocampus of rats during cerebral ischemia/reperfusion injury

Recent studies have shown that tea polyphenols can cross the blood-brain barrier, inhibit apoptosis and play a neuroprotective role against cerebral ischemia. Furthermore, tea polyphenols can decrease DNA damage caused by free radicals. We hypothesized that tea polyphenols repair DNA damage and inhibit neuronal apoptosis during global cerebral ischemia/reperfusion. To test this hypothesis, we employed a rat model of global cerebral ischemia/reperfusion. We demonstrated that intraperitoneal injection of tea polyphenols immediately after reperfusion significantly reduced apoptosis in the hippocampal CA1 region; this effect started 6 hours following reperfusion. Immunohistochemical staining showed that tea polyphenols could reverse the ischemia/reperfusion-induced reduction in the expression of DNA repair proteins, X-ray repair cross-complementing protein 1 and apudnic/apyrimidinic endonuclease/redox factor-1 starting at 2 hours. Both effects lasted at least 72 hours. These experimental findings suggest that tea polyphenols promote DNA damage repair and protect against apoptosis in the brain.

类风湿关节炎患者血清MBL、MASP-2、HsCRP与C_3水平的相关性

目的研究类风湿关节炎(RA)患者血清中甘露糖结合凝集素(MBL)/MBL相关的丝氨酸蛋白酶-2(MASP-2)、Hs CRP和C3水平的相关性。方法收集50例RA患者(活动期和非活动期各25例)和40例健康对照者空腹静脉血,用酶联免疫吸附试验和免疫比浊试验分别检测血清MBL、MASP-2、Hs CRP和C3的水平。结果 RA组活动期和非活动期患者血清中MBL水平均显著低于健康对照组(10.05±2.14 ng/m L vs 26.97±11.78 ng/m L;14.79±3.93 ng/m L vs 26.97±11.78 ng/m L,P<0.05);RA组活动期和非活动期患者血清中MASP-2水平均显著低于健康对照组(75.90±9.21 ng/m L vs 251.78±34.50 ng/m L;107.8±14.90 ng/m L vs251.78±34.50 ng/m L)(P<0.05);RA组活动期和非活动期患者血清中Hs CRP水平显著高于健康对照组(40.53±34.81 mg/L vs1.42±1.70 mg/L;2.97±2.51 mg/L vs 1.42±1.70 mg/L,P<0.05),C3水平3组无差异;双变量Pearson相关性分析,RA患者组MBL与MASP-2含量均呈显著正相关(r=0.550,P=0.001)、与Hs CRP含量呈低度负性相关(r=-0.323,P=0.022);与C3含量无相关(r=0.022,P=0.882)。RA组MASP-2与Hs CRP含量呈低度负性相关(r=-0.453,P=0.001);与C3含量无相关(r=0.049,P=0.738)。经ROC曲线分析,Hs CRP曲线下面积最大(0.844,P=0.001),MASP-2和MBL曲线下面积较小(0.025,P=0.001;0.266,P=0.001),Hs CRP(84%)对RA诊断的敏感性高于MBL(10%)、MASP-2(40%)。结论 MBL与MASP-2水平与Hs CRP呈负相关关系,RA患者关节的炎症损伤程度越重,则血清MBL与MASP-2水平越低,提示MBL与MASP-2参与RA病理损伤过程,但二者的敏感性均较低,不能做为RA诊断及疾病活动程度评价的独立指标。

无水奶油和BL-41的比例对淡奶油稳定性的影响

研究了无水奶油和BL-41两种油脂之间不同的比例对淡奶油粒径分布、界面蛋白含量、脂肪部分聚结率及表观粘度的影响,并在此基础上探讨了其作用机理。研究结果表明,随着BL-41比例的不断增大,淡奶油的上层粒径d3,2、脂肪部分聚结率和表观粘度呈先增大后减小的趋势,在无水奶油∶BL-41为17.5∶17.5时达到最大值,而界面蛋白含量则先降低后升高,在无水奶油∶BL-41为17.5∶17.5时达到最小值。此外,随着储存时间的延长,上层粒径d3,2、脂肪部分聚结率和表观粘度均逐渐增大,而界面蛋白含量则逐渐降低。

布鲁氏菌鞭毛钩蛋白FlgE和融合蛋白BLS-FlgE的克隆、表达及免疫诊断研究

目的分别构建含目的基因flgE和bls-flgE的原核表达载体,表达纯化重组蛋白,并对其免疫诊断价值进行初步评价。方法从NCBI得到布鲁氏菌鞭毛钩蛋白基因flgE和2,4二氧四氢蝶啶合酶基因(bls)序列,设计引物,利用PCR技术扩增得到目的基因,与载体pET30a连接,转化E.coli BL21感受态细胞,IPTG诱导蛋白表达,SDS-PAGE及Western blot鉴定,使用带His标签的蛋白纯化试剂盒纯化蛋白。纯化后的蛋白建立ELISA法检测布病阳性血清和健康人血清,收集灵敏度和特异度数据。结果成功克隆了FlgE鞭毛钩蛋白和BLS-FlgE融合蛋白,经过优化表达条件,目的蛋白获得较大量的表达,免疫印迹结果显示可被阳性病人血清识别。以FlgE蛋白构建ELISA法检测得到灵敏度和特异度分别为70.83%和41.67%,以BLS-FlgE蛋白构建ELISA法检测得到灵敏度和特异度分别为68.75%和52.08%。结论成功表达布鲁氏菌鞭毛钩蛋白FlgE和融合蛋白BLS-FlgE,分别作为诊断抗原总体符合率低,免疫诊断价值较低。

Analysis of immunoglobulin, complements and CRP levels in serum of captive northern pig-tailed macaques （Macaca leonina）

The northem pig-tailed macaque （NPM, Macaca leonina） has become a widely used animal model in biomedical research. In this study, we measured serum immunoglobulin IgG, IgM, IgA, complement C3, C4 and CRP levels in 3-11 year old captive northem pig-tailed macaques using HITACHI 7600-20 automated chemistry analyzer in order to determine the influences of age and gender on these items. The results showed that serum IgA, IgM, C3 and C4 levels were not correlated with age （P〉0.05）, while serum IgG levels increased progressively with age （r=0.202; P=0.045）. Serum IgG, IgA, IgM and C3 levels were higher in females than in males （P〈0.05）. Moreover, serum C3 concentration was both positively and strongly correlated with that of C4 （r=0.700; P〈0.0001）. This study provides basic serum immunoglobulin and complement data of captive northem pig-tailed macaques, which may prove useful for future breeding efforts and biomedical research.

Complements are involved in alcoholic fatty liver disease, hepatitis and fibrosis

The complement system is a key component of the body's immune system. When abnormally activated, this system can induce inflammation and damage to normal tissues and participate in the development and progression of a variety of diseases. In the past, many scholars believed that alcoholic liver disease(ALD) is induced by the stress of ethanol on liver cells, including oxidative stress and dysfunction of mitochondria and protease bodies, causing hepatocyte injury and apoptosis. Recent studies have shown that complement activation is also involved in the genesis and development of ALD. This review focuses on the roles of complement activation in ALD and of therapeutic intervention in complement-activation pathways. We intend to provide new ideas on the diagnosis and treatment of ALD.

Protein corona mediated liposomal drug delivery for bacterial infection management

Liposomes have been widely investigated as a class of promising antibiotic delivery systems for the treatment of life-threatening bacterial infections.However,the inevitable formation of protein corona on the liposomal surface can heavily impact in vivo performance.A better understanding of the effects of protein corona on liposomal behavior can significantly improve antibacterial liposomal drug development.Here,the critical role of protein corona in mediating liposome-bacteria interactions was elucidated.Adsorption of negatively charged protein on cationic liposome weakened electrostatic attractionenhanced liposomal binding to the bacteria.Cumulative complement deposition on anionic liposome composed of phosphatidylglycerol(DSPG s Lip)contributed to a superior binding affinity of DSPG s Lip to planktonic bacteria and biofilms,which was exploited to enhance bacteria-targeted drug delivery.In both S.aureus-related osteomyelitis and pneumonia mice models,DSPG s Lip was demonstrated as a promising antibiotic nanocarrier for managing MRSA infection,indicating the benefits of lipid composition-based protein corona modulation in liposomal antibiotic delivery for bacterial infection treatment.

Cell cycle and complement inhibitors may be specific for treatment of spinal cord injury in aged and young mice: transcriptomic analyses

Previous studies have reported age-specific pathological and functional outcomes in young and aged patients suffering spinal cord injury,but the mechanisms remain poorly understood. In this study, we examined mice with spinal cord injury. Gene expression profiles from the Gene Expression Omnibus database （accession number GSE93561） were used, including spinal cord samples from 3 young injured mice （2–3-months old, induced by Impactor at Th9 level） and 3 control mice （2–3-months old, no treatment）, as well as 2 aged injured mice （15–18-months old, induced by Impactor at Th9 level） and 2 control mice （15–18-months old, no treatment）. Differentially expressed genes （DEGs） in spinal cord tissue from injured and control mice were identified using the Linear Models for Microarray data method,with a threshold of adjusted P 〈 0.05 and ｜logFC（fold change）｜ 〉 1.5. Protein–protein interaction networks were constructed using data from the STRING database, followed by module analysis by Cytoscape software to screen crucial genes. Kyoto encyclopedia of genes and genomes pathway and Gene Ontology enrichment analyses were performed to investigate the underlying functions of DEGs using Database for Annotation, Visualization and Integrated Discovery. Consequently, 1,604 and 1,153 DEGs were identified between injured and normal control mice in spinal cord tissue of aged and young mice, respectively. Furthermore, a Venn diagram showed that 960 DEGs were shared among aged and young mice, while 644 and 193 DEGs were specific to aged and young mice, respectively. Functional enrichment indicates that shared DEGs are involved in osteoclast differentiation, extracellular matrix–receptor interaction, nuclear factor-kappa B signaling pathway, and focal adhesion. Unique genes for aged and young injured groups were involved in the cell cycle （upregulation of PLK1） and complement （upregulation of C3） activation, respectively. These findings were confirmed by functional analysis of genes in modules （common, 4; aged, 2; young, 1） screened from protein–protein interaction networks. Accordingly, cell cycle and complement inhibitors may be specific treatments for spinal cord injury in aged and young mice, respectively.

Neuronal effects of SP600125 pretreatment in a rat model of cerebral ischemia/reperfusion injury Inhibited down-regulation of DNA repair protein

BACKGROUND： Recent studies have shown that the selective inhibitor of c-Jun N-terminal kinases （JNKs） signaling pathway, SP600125, exhibits neuronal protective effects in a rat model of brain ischemia/reperfusion. OBJECTIVE： To determine the mechanisms of neuroprotective effects of SP600125 in a rat model of brain ischemia/reperfusion, and determine the role of the JNK signaling pathway in SP600125-induced effects. DESIGN, TIME AND SETTING： A randomized, controlled, animal experiment was performed at the Animal Experiment Center, Medical School of Xi＇an Jiaotong University from June 2007 to September 2008. MATERIALS： SP600125 was provided by Biosource, USA; rabbit anti-phospho-JNK （Thr183/Tyr185） polyclonal antibody from Cell Signaling Technology, USA; rabbit anti-X-ray repair cross-complementing protein 1 （XRCC1） and anti-Ku70 polyclonal antibodies from Santa Cruz Biotechnology, USA; and TUNEL kit from Beijing Huamei Biology, China. METHODS： A total of 108 male, 4-month-old, Sprague Dawley rats were randomly assigned to three groups, with 36 rats per group. The sham operation group and ischemia/reperfusion group （I/R group） were intracerebroventricularly injected with 10 μL 1% DMSO. The SP600125-treated group （pre-SP group） was given 10 μL SP600125 （3 μg/μL）. Thirty minutes later, brain ischemia was induced in the I/R and pre-SP groups using the four-vessel occlusion method. Specifically, whole brain ischemia was induced for 6 minutes, and the clips were released to restore carotid artery blood flow. Rats from each group were observed at 2, 6, 12, 24, 48, and 72 hours, with 6 rats for each time point. The sham operation group was treated with the same surgical exposure procedures, with exception of occlusion of the carotid artery. MAIN OUTCOME MEASURES： Hematoxylin-eosin staining was used to observe neuronal survival in the hippocampal CA1 region, TUNEL was used to detect apoptosis in the hippocampal CA1 region, and immunohistochemistry was used to detect expression of phospho-JNK, XRCC1, and Ku70. RESULTS： Following brain ischemia/reperfusion, neuronal survival significantly decreased, and the number of apoptotic cells significantly increased （P 〈 0.01）. Compared with the I/R group, neuronal survival significantly increased in the pre-SP group, and the number of apoptotic cells significantly decreased （P 〈 0.01）. Expression of phospho-JNK increased, and XRCC1 and Ku70 significantly decreased （P 〈 0.05） following ischemia/reperfusion. Compared with the I/R group, expression of phospho-JNK decreased, and XRCC1 and Ku70 significantly increased in the pre-SP group （P 〈 0.05）. Correlation analysis revealed an inverse correlation between phospho-JNK gray value and XRCC1 and Ku70 gray values in the hippocampal CA1 region （r = -0.983, -0.953, P 〈 0.01）. CONCLUSION： SP600125 treatment decreased apoptosis induced by global brain ischemia/reperfusion in the rat hippocampal CA1 region. Results suggested that the neuroprotective effects were due to inhibited phosphorylation of JNK and reduced down-regulation of XRCC1 and Ku70.

Expression of CD55, CD59, CD46 and CD35 in Peripheral Blood Cells from Rheumatoid Arthritis Patients

Evidences point that complement activation plays a role in rheumatoid arthritis (RA). In this context, expression of the CReg was investigated on white cells from peripheral blood of 30 RA patients and 30 healthy controls. Using flow cytometric analyses the relative fluorescence intensities (MFI) of Cregs were determined. CD59 MFI was significantly increased in RA cells comparing to controls, respectively: lymphocytes 36.8 versus 27.07;monocytes 32.0 versus 21.37;and granulocytes: 84.6 versus 66.1 (p 0.05). Interestingly, no difference was observed on the MFI to CD55, CD46 and CD35 in these cells. These data indicate an overexpression of CD59 in all the peripheral blood cells of RA patients, perhaps due to an increased synthesis for compensatory mechanisms because of complement activation, inflammatory status or other factors associated with the disease.

Development of Split-Protein Systems: From Binary to Ternary System

Tens of thousands of protein-protein interactions (PPIs) have been found in human cells and many of these macromolecular partnerships could determine the cell growth and death. Thus there is a need to develop the methods to catalogue these macromolecules by detecting their interactions, modifications, and cellular locations. It will be helpful for scientists to compare the difference between a diseased cellular state and its normal state and to find the potential therapy treatment to intervene this status. One technology called split-protein reassembly or protein fragment complementation has been developed in the last two decades. This technology makes use of appropriate fragmentation of some protein reporters and the refolding of these reports could be detected by their function to confirm the interaction of interest. This system has been set up in cell-free systems, E. coli, yeast, mammalian cells, plants and live animals. Herein, I present the development in fluorescence- and bioluminescence-based split-protein biosensors in both binary and ternary systems. In addition, some people developed the split-protein system by combining it with chemical inducer of dimerization strategy (CID). This has been applied for identifying the enzyme inhibitors and regulating the activity of protein kinases and phosphatases. With effort from many laboratories from the world, a variety of split-protein systems have been developed for studying the PPI in vitro and in vivo, monitoring the biological process, and controlling the activity of the enzyme of interest.

Effects of interferon α-2b on liver function, complement level and oxidative stress in patients with hepatitis B

Objective:To observe the effect of interferonα-2b treatment on liver function,liver fibrosis,complement protein and oxidative stress in patients with hepatitis B.Methods:A total of 100 patients with hepatitis B in our hospital were randomly divided into the control group and the observation group,with 50 cases in each group.After admission,patients in the control group were treated with entecavir,while patients in the observation group were treated with interferonα-2b combined with entecavir.The levels of serum total bilirubin(TBil),aspartate aminotransferase(AST),alanine aminotransferase(ALT),type III procollagen(PCIII),type IV collagen(CIV),complement C3 protein(C3),complement C4 protein(C4),malondialdehyde(MDA),superoxide dismutase(SOD)and nitric oxide(NO)were compared between the two groups before and after treatment.Results:After treatment,the levels of TBil,AST,ALT,PCIII,CIV,MDA and NO in serum of patients with hepatitis B in both groups were significantly lower than those before treatment,and the levels of C3,C4 and SOD were significantly higher than those before treatment(P<0.05).After treatment,the levels of TBil,AST,ALT,PC III,C IV,MDA and NO in serum of patients in the observation group were significantly lower than those in the control group,while the levels of C3,C4 and SOD in serum of patients in the observation group were significantly higher than those in the control group(P<0.05).Conclusions:The combination of interferonα-2b and entecavir has a good curative effect on hepatitis B.It can significantly improve the liver function and immune function of patients,delay the process of liver fibrosis and reduce oxidative stress injury.It is worthy of clinical promotion.