Calcium-binding ability of soy protein hydrolysates

This present study investigated the ability of various soy protein hydrolysates (SPHs) in binding calcium. It was demonstrated that the amount of Ca-bound depended greatly on the SPHs obtained using different proteases, which included: neutrase, flavourzyme, protease M and pepsin. The maximum level of Ca-bound (66.9 mg/g) occurred when protease M was used to hydrolyze soy protein. Peptide fragments exhibiting high Ca-binding capacity had molecular weights of either 14.4 or 8–9 kDa. The level of Ca-bound increased linearly with the increment of carboxyl content in SPHs, and further deamidation on SPHs from protease M improved Ca-binding of the hydrolysate.

S100 calcium binding protein A6 and associated long noncoding ribonucleic acids as biomarkers in the diagnosis and staging of primary biliary cholangitis

BACKGROUND Primary biliary cholangitis(PBC)is a chronic and slowly progressing cholestatic disease,which causes damage to the small intrahepatic bile duct by immunoregulation,and may lead to cholestasis,liver fibrosis,cirrhosis and,eventually,liver failure.AIM To explore the potential diagnosis and staging value of plasma S100 calcium binding protein A6(S100A6)messenger ribonucleic acid(mRNA),LINC00312,LINC00472,and LINC01257 in primary biliary cholangitis.METHODS A total of 145 PBC patients and 110 healthy controls(HCs)were enrolled.Among them,80 PBC patients and 60 HCs were used as the training set,and 65 PBC patients and 50 HCs were used as the validation set.The relative expression levels of plasma S100A6 mRNA,long noncoding ribonucleic acids LINC00312,LINC00472 and LINC01257 were analyzed using quantitative reverse transcription-polymerase chain reaction.The bile duct ligation(BDL)mouse model was used to simulate PBC.Then double immunofluorescence was conducted to verify the overexpression of S100A6 protein in intrahepatic bile duct cells of BDL mice.Human intrahepatic biliary epithelial cells were treated with glycochenodeoxycholate to simulate the cholestatic environment of intrahepatic biliary epithelial cells in PBC.RESULTS The expression of S100A6 protein in intrahepatic bile duct cells was up-regulated in the BDL mouse model compared with sham mice.The relative expression levels of plasma S100A6 mRNA,log10 LINC00472 and LINC01257 were upregulated while LINC00312 was down-regulated in plasma of PBC patients compared with HCs(3.01±1.04 vs 2.09±0.87,P<0.0001;2.46±1.03 vs 1.77±0.84,P<0.0001;3.49±1.64 vs 2.37±0.96,P<0.0001;1.70±0.33 vs 2.07±0.53,P<0.0001,respectively).The relative expression levels of S100A6 mRNA,LINC00472 and LINC01257 were up-regulated and LINC00312 was down-regulated in human intrahepatic biliary epithelial cells treated with glycochenodeoxycholate compared with control(2.97±0.43 vs 1.09±0.08,P=0.0018;2.70±0.26 vs 1.10±0.10,P=0.0006;2.23±0.21 vs 1.10±0.10,P=0.0011;1.20±0.04 vs 3.03±0.15,P<0.0001,respectively).The mean expression of S100A6 in the advanced stage(III and IV)of PBC was up-regulated compared to that in HCs and the early stage(II)(3.38±0.71 vs 2.09±0.87,P<0.0001;3.38±0.71 vs 2.57±1.21,P=0.0003,respectively);and in the early stage(II),it was higher than that in HCs(2.57±1.21 vs 2.09±0.87,P=0.03).The mean expression of LINC00312 in the advanced stage was lower than that in the early stage and HCs(1.39±0.29 vs 1.56±0.33,P=0.01;1.39±0.29 vs 2.07±0.53,P<0.0001,respectively);in addition,the mean expression of LINC00312 in the early stage was lower than that in HCs(1.56±0.33 vs 2.07±0.53,P<0.0001).The mean expression of log10 LINC00472 in the advanced stage was higher than those in the early stage and HCs(2.99±0.87 vs 1.81±0.83,P<0.0001;2.99±0.87 vs 1.77±0.84,P<0.0001,respectively).The mean expression of LINC01257 in both the early stage and advanced stage were up-regulated compared with HCs(3.88±1.55 vs 2.37±0.96,P<0.0001;3.57±1.79 vs 2.37±0.96,P<0.0001,respectively).The areas under the curves(AUC)for S100A6,LINC00312,log10 LINC00472 and LINC01257 in PBC diagnosis were 0.759,0.7292,0.6942 and 0.7158,respectively.Furthermore,the AUC for these four genes in PBC staging were 0.666,0.661,0.839 and 0.5549,respectively.The expression levels of S100A6 mRNA,log10 LINC00472,and LINC01257 in plasma of PBC patients were decreased(2.35±1.02 vs 3.06±1.04,P=0.0018;1.99±0.83 vs 2.33±0.96,P=0.036;2.84±0.92 vs 3.69±1.54,P=0.0006),and the expression level of LINC00312 was increased(1.95±0.35 vs 1.73±0.32,P=0.0007)after treatment compared with before treatment using the paired t-test.Relative expression of S100A6 mRNA was positively correlated with log10 LINC00472(r=0.683,P<0.0001);serum level of collagen type IV was positively correlated with the relative expression of log10 LINC00472(r=0.482,P<0.0001);relative expression of S100A6 mRNA was positively correlated with the serum level of collagen type IV(r=0.732,P<0.0001).The AUC for the four biomarkers obtained in the validation set were close to the training set.CONCLUSION These four genes may potentially act as novel biomarkers for the diagnosis of PBC.Moreover,LINC00472 acts as a potential biomarker for staging in PBC.

Interaction of calcium- and integrin-binding protein 1 with integrin <i>α</i>11 and its possible involvement in pulmonary fibrosis

Integrin α11 (ITGA11) is one of the collagen-binding integrin α chains;however, its biological significance remains unknown. To determine the functions of ITGA11, we performed a yeast two-hybrid screen using the cytoplasmic domain of ITGA11 as bait and transformed an EGY48 yeast strain with the bait-containing plasmid using the plasmid from a human lung fibroblast cDNA library. This screen identified calcium- and integrin-binding protein 1 (CIB1) as prey. Recombinant ITGA11 and CIB1 were expressed in mammalian cells and used in coimmunoprecipitation experiments, which showed that full-length ITGA11 and CIB1 are also associated in vivo. Over-expression of CIB1 in the human lung myofibroblast MRC-5 cells decreased the expression of α-smooth muscle actin and fibronectin. Using a mouse model of pulmonary fibrosis (bleomycin-treatment), we detected elevated expression of CIB1 in lung tissues compared with controls. These data suggest that CIB1 may regulate pulmonary fibrosis in concert with IT-GA11.

Knockdown of polypyrimidine tract binding protein facilitates motor function recovery after spinal cord injury

After spinal cord injury(SCI),a fibroblast-and microglia-mediated fibrotic scar is formed in the lesion core,and a glial scar is formed around the fibrotic scar as a res ult of the activation and proliferation of astrocytes.Simultaneously,a large number of neuro ns are lost in the injured area.Regulating the dense glial scar and re plenishing neurons in the injured area are essential for SCI repair.Polypyrimidine tra ct binding protein(PTB),known as an RNA-binding protein,plays a key role in neurogenesis.Here,we utilized short hairpin RNAs(shRNAs)and antisense oligonucleotides(ASOs)to knock down PTB expression.We found that reactive spinal astrocytes from mice were directly reprogrammed into motoneuron-like cells by PTB downregulation in vitro.In a mouse model of compressioninduced SCI,adeno-associated viral shRNA-mediated PTB knockdown replenished motoneuron-like cells around the injured area.Basso Mouse Scale scores and forced swim,inclined plate,cold allodynia,and hot plate tests showed that PTB knockdown promoted motor function recovery in mice but did not improve sensory perception after SCI.Furthermore,ASO-mediated PTB knockdown improved motor function resto ration by not only replenishing motoneuron-like cells around the injured area but also by modestly reducing the density of the glial scar without disrupting its overall structure.Together,these findings suggest that PTB knockdown may be a promising therapeutic strategy to promote motor function recovery during spinal cord repair.

Novel Treatment Approach in Schizophrenia: Substitution of Glial Binding Proteins

In chronic schizophrenia, synaptic information processing is unbalanced, as shown in a model of glial-neuronal synaptic units, called tripartite synapses. The glial component of the synapse exerts a modifying function in neurotransmission since the astrocyte activated by neurotransmitters produces gliotransmitters that negatively feedback to the presynapse. It is hypothesized that in schizophrenia nonfunctional astrocytic receptors cannot be activated, thus losing their modulating function. This causes a generalization of information processing in the neuronal networks such that the brain is unable to distinguish between subjects and objects in the environment. Delusions, hallucinations and cognitive impairment occur on the behavioral level. In a model of a cholinergic tripartite synapse, it is shown that glial binding proteins modify neurotransmission by occupancy with cognate neurotransmitters temporarily turning off neurotransmission on the presynapse. Most recently, glial binding proteins have been engineered. It is proposed that the substitution of glial binding proteins may balance synaptic information processing in schizophrenia since these proteins exert a modulatory function comparable to functional astrocytic receptors. Rap- id technical developments may enable this novel treatment approach in schizophrenia.

Endoplasmic reticulum stress transducer old astrocyte specifically induced substance contributes to astrogliosis after spinal cord injury

Old astrocyte specifically induced substance （OASIS） is an endoplasmic reticulum （ER） stress transducer specifically expressed in astrocytes and osteoblasts. OASIS regulates the differentiation of neural precursor cells into astrocytes in the central nervous system. This study aimed to elucidate the involvement of ER stress responses stimulated via OASIS in astrogliosis following spinal cord injury. In a mouse model of spinal cord contusion injury, OASIS mRNA and protein expression were evaluated at days 7 and 14. A significant increase in OASIS mRNA on day 7 and an increase in protein on days 7 and 14 was observed in injured spinal cords. Immunostaining on day 7 revealed co-localization of OASIS and astrocytes in the periphery of the injury site. Furthermore, anti-OASIS small interfering RNA （siRNA） was injected at the injury sites on day 5 to elucidate the function of OASIS. Treatment with anti-OASIS siRNA caused a significant decrease in OASIS mRNA on day 7 and protein on days 7 and 14, and was associated with the inhibition of astrogliosis and hindlimb motor function recovery. Results of our study show that OASIS expression synchronizes with astrogliosis and is functionally associated with astrogliosis after spinal cord injury.

Edaravone protects against oxygen-glucose-serum deprivation/restoration-induced apoptosis in spinal cord astrocytes by inhibiting integrated stress response

We previously found that oxygen-glucose-serum deprivation/restoration（OGSD/R） induces apoptosis of spinal cord astrocytes, possibly via caspase-12 and the integrated stress response, which involves protein kinase R-like endoplasmic reticulum kinase（PERK）, eukaryotic initiation factor 2-alpha（eIF2α） and activating transcription factor 4（ATF4）. We hypothesized that edaravone, a low molecular weight, lipophilic free radical scavenger, would reduce OGSD/R-induced apoptosis of spinal cord astrocytes. To test this, we established primary cultures of rat astrocytes, and exposed them to 8 hours/6 hours of OGSD/R with or without edaravone（0.1, 1, 10, 100 μM） treatment. We found that 100 μM of edaravone significantly suppressed astrocyte apoptosis and inhibited the release of reactive oxygen species. It also inhibited the activation of caspase-12 and caspase-3, and reduced the expression of homologous CCAAT/enhancer binding protein, phosphorylated（p）-PERK, p-eIF2α, and ATF4. These results point to a new use of an established drug in the prevention of OGSD/R-mediated spinal cord astrocyte apoptosis via the integrated stress response.

Immortalized hippocampal astrocytes from 3xTg-AD mice,a new model to study disease-related astrocytic dysfunction:a comparative review

Alzheimer's disease(AD)is characterized by complex etiology,long-lasting pathogenesis,and celltype-specific alterations.Currently,there is no cure for AD,emphasizing the urgent need for a comprehensive understanding of cell-specific pathology.Astrocytes,principal homeostatic cells of the central nervous system,are key players in the pathogenesis of neurodegenerative diseases,including AD.Cellular models greatly facilitate the investigation of cell-specific pathological alterations and the dissection of molecular mechanisms and pathways.Tumor-derived and immortalized astrocytic cell lines,alongside the emerging technology of adult induced pluripotent stem cells,are widely used to study cellular dysfunction in AD.Surprisingly,no stable cell lines were available from genetic mouse AD models.Recently,we established immortalized hippocampal astroglial cell lines from amyloid-βprecursor protein/presenilin-1/Tau triple-transgenic(3xTg)-AD mice(denominated as wild type(WT)-and 3Tg-iAstro cells)using retrovirus-mediated transduction of simian virus 40 large T-antigen and propagation without clonal selection,thereby maintaining natural heterogeneity of primary cultures.Several groups have successfully used 3Tg-iAstro cells for single-cell and omics approaches to study astrocytic AD-related alterations of calcium signaling,mitochondrial dysfunctions,disproteostasis,altered homeostatic and signaling support to neurons,and blood-brain barrier models.Here we provide a comparative overview of the most used models to study astrocytes in vitro,such as primary culture,tumor-derived cell lines,immortalized astroglial cell lines,and induced pluripotent stem cell-derived astrocytes.We conclude that immortalized WT-and 3Tg-iAstro cells provide a noncompetitive but complementary,low-cost,easy-to-handle,and versatile cellular model for dissection of astrocyte-specific AD-related alterations and preclinical drug discovery.

电针对大鼠局灶性脑缺血脑内Calbindin-D_(28k)表达的影响

目的 :探讨钙结合蛋白家族成员Calbindin D2 8k与电针抗缺血性脑损伤的关系。方法 :用改良线栓法制备SD大鼠大脑中动脉 (MCA)阻塞 再灌注模型 ,应用TTC及HE组织化学染色观察缺血及电针后梗塞灶的变化 ,并应用免疫组织化学方法对大鼠局灶性脑缺血组织内Calbindin D2 8k免疫阳性细胞的分布及电针对该分布的影响进行观察。结果 :电针可明显改善局灶性脑缺血所造成的神经缺损体征 ;但缺血组及电针组Calbindin D2 8k免疫阳性细胞在梗塞灶中心区几乎检测不出 ,缺血组在半影区与对照组相比表达上调 (P <0 0 5 ) ;电针组Calbindin D2 8k在半影区的表达与缺血组之间差异无显著性意义 (P >0 0 5 )。结论 :局灶性脑缺血对Calbindin D2 8k在半影区反应性表达增强可能用以缓冲胞内过多的游离钙 ;电针在脑缺血时对神经元具有保护效应 ,但可能并不是通过调整Calbindin D2

人胎小脑内Calbind in和Parvalbum in的免疫组织化学研究(英文)

既往研究显示的哺乳动物小脑单位和小脑缝隙，在人类称之为Ｐｕｒｋｉｎｊｅ 细胞群和缝隙。进一步的研究证实，一种钙结合蛋白Ｃａｌｂｉｎｄｉｎ（ＣＢ）在啮齿动物小脑的发育过程中可作为Ｐｕｒｋｉｎｊｅ 细胞群的短时标志物。为了观察人胎Ｐｕｒｋｉｎｊｅ 细胞群和小脑缝隙的发育变化，并试图证实在小脑Ｐｕｒｋｉｎｊｅ 细胞的发育过程中钙结合蛋白的表达是否有差别，本文选用胎龄１３、１５、２０、２６、３２和３９ 周的小脑分别作ＣＢ和Ｐａｒｖａｌｂｕｍ ｉｎ（ＰＶ）的免疫组织化学反应，ＣＢ和ＰＶ 均显示内外方向的空间分布顺序，并且除１３ 周ＣＢ有表达而ＰＶ 没有表达外，其它胎龄均显示了相似的分布模式。在第１３ 周，ＣＢ阳性的Ｐｕｒｋｉｎｊｅ 细胞前体仅见于小脑蚓，而小脑半球则不存在，阳性细胞体和轴突组合成前后方向的细胞群，与非免疫反应性的条带交替出现。至第１５ 周，小脑蚓和半球的Ｐｕｒｋｉｎｊｅ 细胞均可被ＣＢ和ＰＶ 标记。ＣＢ和ＰＶ 阳性胞体和轴突以非免疫反应的缝隙为界，在小脑蚓和蚓旁组织组成纵向的细胞群。小脑叶片内Ｐｕｒｋｉｎｊｅ 细胞群和缝隙在前后和背腹方向上排列一致。小脑半球内的Ｐｕｒｋｉｎｊｅ 细胞缺乏这种条带状结构。从２４周起。

大鼠食管肌间神经元nNOS与calbindin的共存及其与衰老关系的研究(英文)

本实验通过观察一氧化氮合酶(nNOS)与细胞内钙结合蛋白(calbindin, CB)共存情况,旨在研究大鼠食管内NO类神经元的神经化学特性并试图寻找出引起食管NO类神经元在衰老过程中丢失的可能原因.取自不同年龄组及种系(Wistar和S-D)大鼠的食管组织,制成肌间神经丛铺片,经nNOS与CB免疫组化双重染色后在荧光显微镜下观察.结果显示,nNOS与CB的免疫阳性反应物均见于大鼠食管肌间神经元胞体内.根据二者共存情况,大鼠食管内nNOS免疫阳性神经元可分为两大亚类:nNOS+/CB+及nNOS+/CB-.而且我们首次发现在大鼠食管腹腔段含有大量nNOS+/CB+神经元,约占NO类神经元总体的30%～40%,与食管其它部位相比有显著差异(P<0.001).另外,大鼠食管腹腔段绝大多数CB免疫阳性神经元同时也为nNOS免疫阳性(约占95%～100%).上述特征均见于本实验所观察的所有不同年龄及不同种系的大鼠食管内.在大鼠衰老过程中,nNOS+/CB+神经元的相对百分比的变化具有种系特异性.线形回归实验分析表明:年轻SD大鼠食管内nNOS+/CB+神经元的百分比含量与衰老过程中NO类神经元丢失数量的百分比之间存在显著负相关性(correlation coefficient 0.99; P<0.05).结果提示在S-D大鼠衰老过程中其食管内NO类神经元的丢失可能与其细胞内钙结合蛋白含量的变化有关.但nNOS/CB共存神经元在分布上有明显差异的生物学意义尚有待于进一步研究.

大鼠下丘脑内Calbindin-D28K和NADPH-d的共存

应用免疫细胞化学方法结合NADPH－d组织化学技术对大鼠下丘脑内calbindin－D28K和NADPH－d的共存进行了实验研究。结果发现：在视上核、下丘脑室旁核大细胞部可见到较多的calbindin－D28K／NADPH－d双重反应阳性神经元．这些神经元分别占以上两核团神经元总数的52．8％和42．5％；而在下丘脑的其它部位如下丘脑外侧区、下丘脑腹内侧核、穹隆周核和内侧视前区等处也可见到少量的双重反应神经元、本文的研究结果表明calbindin－D28K和NADPH－d共存于下丘脑的单个神经元内。

Calbindin D28k对帕金森病模型小鼠纹状体凋亡相关蛋白表达的影响

目的观察1-甲基4-苯基1,2,3,6四氢吡啶(MPTP)致帕金森病(PD)模型小鼠黑质多巴胺能神经细胞过表达Calbindin D28k(CB)时,纹状体细胞抗损伤作用机制。方法选择C57BL/6小鼠连续5 d腹腔注射MPTP,构建成功PD模型小鼠30只,随机分为模型组,人类免疫缺陷病毒(HIV)Ⅰ组(注射HIV Ⅰ)和CB-HIV-Ⅰ组(注射CB-HIV-Ⅰ),每组10只,连续6周对各组小鼠行为学检测,Western blot法检测各组小鼠CB,Bcl 2和Bax的表达变化。结果与模型组和HIV-Ⅰ组比较,CB-HIV-Ⅰ组小鼠各时间点移动格子次数,第1、2、5和6周站立次数,第6周时游泳和悬挂时间,差异有统计学意义(P<0.05,P<0.01);CB-HIV Ⅰ组小鼠中脑黑质中CB的表达量显著升高(P<0.05),纹状体细胞中Bcl-2的表达量亦明显升高(P<0.01),而Bax的表达量明显降低(P<0.01)。模型组和HIV-Ⅰ组上述指标差异无统计学意义(P>0.05)。结论黑质多巴胺能神经细胞过表达CB时,纹状体细胞Bcl-2/Bax表达上调,提示其与纹状体细胞抗凋亡能力增强有关。

Targeting GATA1 and p2x7r Locus Binding in Spinal Astrocytes Suppresses Chronic Visceral Pain by Promoting DNA Demethylation

Irritable bowel syndrome is a gastrointestinal disorder of unknown etiology characterized by widespread,chronic abdominal pain associated with altered bowel movements.Increasing amounts of evidence indicate that injury and inflammation during the neonatal period have long-term effects on tissue structure and function in the adult that may predispose to gastrointestinal diseases.In this study we aimed to investigate how the epigenetic regulation of DNA demethylation of the p2x7r locus guided by the transcription factor GATA binding protein 1(GATA1)in spinal astrocytes affects chronic visceral pain in adult rats with neonatal colonic inflammation(NCI).The spinal GATA1 targeting to DNA demethylation of p2x7r locus in these rats was assessed by assessing GATA1 function with luciferase assay,chromatin immunoprecipitation,patch clamp,and interference in vitro and in vivo.In addition,a decoy oligodeoxynucleotide was designed and applied to determine the influence of GATA1 on the DNA methylation of a p2x7r CpG island.We showed that NCI caused the induction of GATA1,Ten-eleven translocation 3(TET3),and purinergic receptors(P2X7Rs)in astrocytes of the spinal dorsal horn,and demonstrated that inhibiting these molecules markedly increased the pain threshold,inhibited the activation of astrocytes,and decreased the spinal sEPSC frequency.NCI also markedly demethylated the p2x7r locus in a manner dependent on the enhancement of both a GATA1–TET3 physical interaction and GATA1 binding at the p2x7r promoter.Importantly,we showed that demethylation of the p2x7r locus(and the attendant increase in P2X7R expression)was reversed upon knockdown of GATA1 or TET3 expression,and demonstrated that a decoy oligodeoxynucleotide that selectively blocked the GATA1 binding site increased the methylation of a CpG island in the p2x7r promoter.These results demonstrate that chronic visceral pain is mediated synergistically by GATA1 and TET3 via a DNA-demethylation mechanism that controls p2x7r transcription in spinal dorsal horn astrocytes,and provide a potential therapeutic strategy by targeting GATA1 and p2x7r locus binding.

Calciomics:prediction and analysis of EF-hand calcium binding proteins by protein engineering

Ca2+ plays a pivotal role in the physiology and biochemistry of prokaryotic and mammalian organisms.Viruses also utilize the universal Ca2+ signal to create a specific cellular environment to achieve coexistence with the host,and to propagate.In this paper we first describe our development of a grafting approach to understand site-specific Ca2+ binding properties of EF-hand proteins with a helix-loop-helix Ca2+ binding motif,then summarize our prediction and identification of EF-hand Ca2+ binding sites on a genome-wide scale in bacteria and virus,and next report the application of the grafting approach to probe the metal binding capability of predicted EF-hand motifs within the streptococcal hemoprotein receptor(Shr) of Streptococcus pyrogenes and the nonstructural protein 1(nsP1) of Sindbis virus.When methods such as the grafting approach are developed in conjunction with prediction algorithms we are better able to probe continuous Ca2+-binding sites that have been previously underrepresented due to the limitation of conventional methodology.

Diagnostic and Predictive Levels of Calcium-binding Protein A8 and Tumor Necrosis Factor Receptor-associated Factor 6 in Sepsis-associated Encephalopathy： A Prospective Observational Study

Background： Despite its high prevalence, morbidity, and mortality, sepsis-associated encephalopathy （SAE） is still poorly understood. The aim of this prospective and observational study was to investigate the clinical significance of calcium-binding protein A8 （S 100AS） in serum and tumor necrosis factor receptor-associated factor 6 （TRAF6） in peripheral blood mononuclear cells （PBMCs） in diagnosing SAE and predicting its prognosis. Methods： Data of septic patients were collected within 24 h after Intensive Care Unit admission fi-om July 2014 to March 2015. Healthy medical personnel served as the control group. SAE was defined as cerebral dysfhnction in the presence of sepsis that fulfilled the exclusion criteria. The biochemical indicators, Glasgow Coma Scale, Acute Physiology and Chronic Health Evaluation score II, TRAF6 in PBMC, serum S 100A8, S 10013, and neuron-specific enolase were evaluated in SAE patients afresh. TRAF6 and S 100A8 were also measured in the control group. Results： Of the 57 enrolled patients, 29 were diagnosed with SAE. The S 100A8 and TRAF6 concentrations in SAE patients were both significantly higher than that in no-encephalopathy （NE） patients, and higher in NE than that in controls （3.74 ± 3.13 vs. 1.08 ± 0.75 vs. 0.37 ± 0.14 ng/ml, P 〈 0.01 ; 3.18 ± 1.55 vs. 1.02 ± 0.63 vs. 0.47 ± 0.10, P 〈 0.01）. S 100A8 levels of 1.93 ng/ml were diagnostic of SAE with 92.90% specificity and 69.00% sensitivity in the receiver operating characteristic （ROC） curve, and the area under the curve was 0.86 （95% confidence interval [CI]： 0.76-0.95）. TRAF6-relative levels of 1.44 were diagnostic of SAE with 85.70% specificity and 86.20% sensitivity, and the area under the curve was 0.94 （95% CI： 0.88-0.99）. In addition, S 100A8 levels of 2.41 ng/ml predicted 28-day mortality of SAE with 90.00% specificity and 73.70% sensitivity in the ROC curve, and the area under the curve was 0.88. TRAF6 relative levels of 2.94 predicted 28-day mortality of SAE with 80.00% specificity and 68.40% sensitivity, and the area under the curve was 0.77. Compared with TRAF6, the specificity of serum S 100A8 in diagnosing SAE and predicting mortality was higher, although the sensitivity was low. In contrast, the TRAF6 had higher sensitivity for diagnosis. Conclusions： Peripheral blood levels of S 100A8 and TRAF6 in SAE patients were elevated and might be related to the severity of SAE and predict the outcome of SAE. The efficacy and specificity of S 100A8 for SAE diagnosis were superior, despite its weak sensitivity. S100A8 might be a better biomarker for diagnosis of SAE and predicting prognosis.

Glial fibrillary acidic protein levels are associated with global histone H4 acetylation after spinal cord injury in rats

Emerging evidence has suggested global histone H4 acetylation status plays an important role in neural plasticity. For instance, the imbalance of this epigenetic marker has been hypothesized as a key factor for the development and progression of several neurological diseases. Likewise, astrocytic reactivity-a wellknown process that markedly influences the tissue remodeling after a central nervous system injury-is crucial for tissue remodeling after spinal cord injury（SCI）. However, the linkage between the above-mentioned mechanisms after SCI remains poorly understood. We sought to investigate the relation between both glial fibrillary acidic protein（GFAP） and S100 calcium-binding protein B（S100B）（astrocytic reactivity classical markers） and global histone H4 acetylation levels. Sixty-one male Wistar rats（aged ~3 months） were divided into the following groups： sham; 6 hours post-SCI; 24 hours post-SCI; 48 hours post-SCI; 72 hours post-SCI; and 7 days post-SCI. The results suggested that GFAP, but not S100B was associated with global histone H4 acetylation levels. Moreover, global histone H4 acetylation levels exhibited a complex pattern after SCI, encompassing at least three clearly defined phases（first phase： no changes in the 6, 24 and 48 hours post-SCI groups; second phase： increased levels in the 72 hours post-SCI group; and a third phase： return to levels similar to control in the 7 days post-SCI group）. Overall, these findings suggest global H4 acetylation levels exhibit distinct patterns of expression during the first week post-SCI, which may be associated with GFAP levels in the perilesional tissue. Current data encourage studies using H4 acetylation as a possible biomarker for tissue remodeling after spinal cord injury.

Co-existence of calcium-binding proteins and γ-aminobutyric acid or glycine in neurons of the rat medullary dorsal horn

Background We investigated the co-expression of calb indin-D28k (CB), calretinin (CR) and parvalbumin (PV, a combination of the three is referred to as CaBPs) with γ-aminobutyric acid (GABA) or glycine in neurons of the rat medullary dorsal horn (MDH).Methods Immunofluorescence histochemical double-staining for CaBPs and GABA or glycine was performed on the sections from rat MDH. Results CB-, CR-, PV-, GABA- and glycine-like immunoreactive (LI) neurons were differentially observed in all layers of the MDH, but particularly in lamina Ⅱ. Neurons that exhibited immunoreactivity for both CaBPs and GABA or glycine were also observed mainly in lamina Ⅱ. A few of them were found in laminae I and III. The percentages of neurons which co-expressed CB/GABA or CB/glycine out of the total numbers of CB- and GABA-LI neurons or CB- and glycine-LI neurons were 5.3% and 12.1% or 4.1% and 10.0%, respectively. The ratios of CR/GABA or CR/glycine co-existing neurons out of the total numbers of CR- and GABA-LI neurons or CR- and glycine-LI neurons were 5.8% and 7.6% or 4.4% and 7.1%, respectively. The rates of PV/GABA or PV/glycine co-localized neurons out of the total numbers of PV- and GABA-LI neurons or PV- and glycine-LI neurons were 11.1% and 5.1% or 9.9% and 5.1%, respectively. Conclusion The results indicate that some neurons in the MDH contain both CaBPs and GABA or glycine.

DISTRIBUTION OF PARVALBUMIN, CALBINDIN-D28 AND CALRETININ IMMUNOREACTIVE NEURONS AND FIBERS IN THE MONKEY BASAL GANGLIA

Objective To investigate the cellular localization of parvalbumin (PV), calbindin-D28k (CB) and calretinin (CR) in the monkey basal ganglia.Methods Immunocytochemical technique was used to detect PV,CB and CR immunoreactivity in the basal ganglia. Results In the striatum, CB labeled medium-sized spiny projection neurons whereas PV and CR marked two separate classes of aspiny interneurons. The striatal matrix compartment was markedly enriched with CB while striatal patches displayed a CR-rich neuropil. In the pallidum, virtually all neurons contained PV but none express CB. CR occured only in a small subpopulation of large and small pallidal neurons. In the subthalamic nucleus, there existed a multitude of PV-positive cells and fibers but the number of CR and CB-positive neuronal elements was small. In the substantia nigra / ventral tegmental area complex, CB and CR occured principally in dopaminergic neurons of the dorsal tier of the pars compacta and in those of the ventral tegmental area. PV was strickly confined to the GABAergic neurons of the pars reticular and lateralis. CB-rich fibers abounded in the pars reticular and lateralis, while CR-positive axons were confined to the pars compacta. Conclusion CB and PV were distributed according to a strikingly complementary pattern in primate basal ganglia, and the use of CB and PV immunocytochemistry may be considered as an excellent tool to define distinct chemoarchitectonic and functional domains within the complex organization of the basal ganglia. CR was less ubiquitous but occured in small basal ganglia components where it labeled distinct subsets of neurons. Such highly specific patterns of distribution indicate that CB, PV and CR may work in synery within primate basal ganglia.

血清S100A11、S100A14、CA125与ⅡB~Ⅳ期上皮性卵巢癌患者减瘤术结局的关系及其预测价值分析

目的 探讨ⅡB~Ⅳ期上皮性卵巢癌(EOC)患者血清S100钙结合蛋白A(S100A)11、S100A14和糖类抗原125(CA125)水平与肿瘤细胞减灭术(简称减瘤术)结局的关系及其预测价值。方法 选取2018年6月至2021年6月于该院接受初始减瘤术的ⅡB~Ⅳ期的124例EOC患者作为EOC组及70例卵巢良性病变患者作为病例对照组,并选取同期于该院体检的体检健康者70例作为健康对照组。检测各组血清S100A11、S100A14、CA125水平。根据EOC患者减瘤术结局是否满意,分为满意组(70例)和不满意组(54例)。分析ⅡB~Ⅳ期EOC患者减瘤术结局不满意的影响因素,并评估血清S100A11、S100A14、CA125预测ⅡB~Ⅳ期EOC患者减瘤术结局不满意的临床价值。结果 EOC组血清S100A11、S100A14、CA125水平高于病例对照组及健康对照组,差异有统计学意义(P<0.05)。不满意组美国麻醉医师协会(ASA)分级Ⅲ级、腹水、国际妇产科联盟(FIGO)分期ⅢC~Ⅳ期比例及血清CA125、S100A11、S100A14水平均高于满意组,差异有统计学意义(P<0.05)。FIGO分期ⅢC~Ⅳ期及血清CA125、S100A11、S100A14水平升高是影响EOC患者减瘤术结局不满意的独立危险因素(P<0.05)。血清CA125、S100A11、S100A14及三者联合预测EOC患者减瘤术结局不满意的曲线下面积(95%CI)分别为0.727(0.521~0.910)、0.747(0.507~0.961)、0.755(0.553~0.954)、0.825(0.743~0.913),三者联合检测预测减瘤术结局不满意的价值优于血清CA125、S100A11、S100A14单独检测。结论 ⅡB~Ⅳ期EOC患者血清S100A11、S100A14、CA125水平升高,与减瘤术结局不满意有关,血清CA125、S100A11、S100A14联合检测对减瘤术结局不满意具有较高的预测价值。