SsdchA is a novel secretory cellobiohydrolase driving pathogenicity in Sclerotinia sclerotiorum

The necrotrophic fungus, Sclerotinia sclerotiorum, employs an array of cell wall-degrading enzymes(CWDEs), including cellulase, to dismantle host cell walls. However, the molecular mechanisms through which S. sclerotiorum degrades cellulose remain elusive. Here, we unveil a novel secretory cellobiohydrolase, SsdchA, characterized by a signal peptide and a Glyco_hydro_7(GH7) domain. SsdchA exhibits a robust expression of during early infection stages. Interestingly, colony morphology and growth rates remain unaffected across the wild-type, SsdchA deletion strains and SsdchA overexpression strains on potato dextrose agar(PDA) medium. Nevertheless, the pathogenicity and cellobiohydrolase activity decreased in the SsdchA deletion strains, but enhanced in the SsdchA overexpression strains. Moreover,the heterologous expression of SsdchA in Arabidopsis thaliana leads to reduced cellulose content and heightened susceptibility to S. sclerotiorum. Collectively, our data underscore the pivotal role of the novel cellobiohydrolase SsdchA in the pathogenicity of S. sclerotiorum.

Paracrine and endocrine actions of bone——the functions of secretory proteins from osteoblasts, osteocytes, and osteoclasts

The skeleton is a dynamic organ that is constantly remodeled. Proteins secreted from bone cells, namely osteoblasts, osteocytes,and osteoclasts exert regulation on osteoblastogenesis, osteclastogenesis, and angiogenesis in a paracrine manner. Osteoblasts secrete a range of different molecules including RANKL/OPG, M-CSF, SEMA3A, WNT5A, and WNT16 that regulate osteoclastogenesis. Osteoblasts also produce VEGFA that stimulates osteoblastogenesis and angiogenesis. Osteocytes produce sclerostin（SOST） that inhibits osteoblast differentiation and promotes osteoclast differentiation. Osteoclasts secrete factors including BMP6, CTHRC1, EFNB2, S1P, WNT10B, SEMA4D, and CT-1 that act on osteoblasts and osteocytes, and thereby influencea A osteogenesis. Osteoclast precursors produce the angiogenic factor PDGF-BB to promote the formation of Type H vessels, which then stimulate osteoblastogenesis. Besides, the evidences over the past decades show that at least three hormones or ＂osteokines＂from bone cells have endocrine functions. FGF23 is produced by osteoblasts and osteocytes and can regulate phosphate metabolism. Osteocalcin（OCN） secreted by osteoblasts regulates systemic glucose and energy metabolism, reproduction, and cognition. Lipocalin-2（LCN2） is secreted by osteoblasts and can influence energy metabolism by suppressing appetite in the brain.We review the recent progresses in the paracrine and endocrine functions of the secretory proteins of osteoblasts, osteocytes, and osteoclasts, revealing connections of the skeleton with other tissues and providing added insights into the pathogenesis of degenerative diseases affecting multiple organs and the drug discovery process.

Participation of epididymal cysteine-rich secretory proteins in sperm-egg fusion and their potential use for male fertility regulation

Rat protein DE is an androgen-dependent cysteine-rich secretory protein （CRISP） synthesized by proximal epididymal regions. DE, also known as CRISP-1, is localized on the equatorial segment of acrosome-reacted spermatozoa and participates in gamete fusion through binding to egg complementary sites. Immunization of rats with DE inhibits fertility and sperm fusion ability, suggesting that DE represents a good epididymal contraceptive target. Recombinant DE fragments and synthetic peptides revealed that DE binds to the egg via a 12-amino acid region of an evolutionarily conserved motif, Signature 2 （S2）. The ability of other CRISP to bind to the rat egg was correlated with their S2 amino acid sequences. Although testicular protein Tpx- 1 （CRISP-2） was capable of binding to rodent eggs, human epididymal AEG-related protein （ARP） and helothermine （from lizard saliva） were not. The S2 region presented only two substitutions in Tpx-1 and four in ARP and helothermine, compared with the DE S2, suggesting that this amino acid sequence was relevant for egg interaction. Studies with Tpx- 1 and anti-Tpx- 1 revealed the participation of this protein in gamete fusion through binding to complementary sites in the egg. In competition studies, DE reduced binding of Tpx- 1 dose-dependently, indicating that both CRISP share the egg complementary sites. That anti-DE and anti-Tpx-1 inhibit sperm-egg fusion while recognizing only the corresponding proteins, suggests functional cooperation between these homologous CRISP to ensure fertilization success. These results increase our understanding of the molecular mechanisms of gamete fusion and contribute to the development of new and safer fertility regulating methods. （Asian J Androl 2007 July; 9： 528-532）

Expression of serum human epididymal secretory protein E4 at low grade and high grade serous carcinomas

Objective:To investigate the value of serum human epididymis protein 4(HE4) in differential diagnosis of patients with low-grade serous(LGSC) and high-grade serous carcinoma(HGSC) serous ovarian cancer.Methods:LGSC and HGSC serous ovarian cancer were diagnosed by the two-tier grade system,serum levels of HE4 and carbohydrate antigen 12S(CA125) were measured by ELBA and radioisotope method,respectively in 60 serous ovarian cancer patients. HE4 and TPS3 protein in cancer tissue were measured by immunohistochemical method. Results:The difference in density of HE4 and TP53 protein was significant between LGSC and HGSC tissue,while serum CA12S did not show significant difference between different serum samples.There was significant difference in serum HE4 levels between LGSC and HGSC and the result was different within FIGO(Ⅰ+Ⅱ) stage,suggesting HE4 was not a reliable biomarker for the discrimination between LGSC and HCSC.HE4 had potential as a biomarker for the discrimination between LGSC and HGSC but the role in early diagnosis was limited.Conclusions:HE4 may be a reliable marker for differential diagnosis of LGSC and HGSC.But its role in early diagnosis of LGSC and HGSC need to be confirmed from the perspective of two-tier grade system.

Gene expression profiling defined pathways correlated with fibroblast cell proliferation induced by Opisthorchis viverrini excretory /secretory product

AIM： To investigate the mechanism of fibroblast cell proliferation stimulated by the Opisthorchis viverrini excretory/secretory （ES） product. METHODS： NIH-3T3, mouse fibroblast cells were treated with O. viverrini ES product by non-contact co-cultured with the adult parasites. Total RNA from NIH-3T3 treated and untreated with O. viverrini was extracted, reverse transcribed and hybridized with the mouse 15K complementary DNA （cDNA） array. The result was analyzed by ArrayVision version 5 and GeneSpring version 5 softwares. After normalization, the ratios of gene expression of parasite treated to untreated NIH-3T3 cells of 2-and more-fold upregulated was defined as the differentially expressed genes. The expression levels of the signal transduction genes were validated by semiquantitative SYBR-based real-time RT-PCR. RESULTS： Among a total of 15 000 genes/ESTs, 239 genes with established cell proliferation-related function were 2 fold-and more-up-regulated by O. viverrini ES product compared to those in cells without exposure to the parasitic product. These genes were classified into groups including energy and metabolism, signal transduction, protein synthesis and translation, matrix and structural protein, transcription control, cell cycle and DNA replication. Moreover, the expressions of serinethreonine kinase receptor, receptor tyrosine kinase and collagen production-related genes were up-regulated by O.viverrini ES product, The expression level of signal transduction genes, pkC, pdgfra, jak 1, eps 8, tgfβ 1/4,strap and h ras measured by real-time RT-PCR confirmed their expression levels to those obtained from cDNA array. However, only the up-regulated expression of pkC,eps 8 and tgfβ3 1/4 which are the downstream signaling molecules of either epidermal growth factor （EGF） or transforming growth factor-β （TGF-β） showed statistical significance （P 〈 0.05）.CONCLUSION： O. viverrini ES product stimulates the significant changes of gene expression in several functional categories and these mainly include transcripts related to cell proliferation. The TGF-β and EGF signal transduction pathways are indicated as the possible pathways of O. viverrini-driven cell proliferation.

Secretory Transactivating Transcription-apoptin fusion protein induces apoptosis in hepatocellular carcinoma HepG2 cells

AIM:To determine whether SP-TAT-apoptin induces apoptosis and also maintains its tumor cell specificity. METHODS:In this study,we designed a secretory protein by adding a secretory signal peptide(SP) to the N terminus of Transactivating Transcription(TAT)-apoptin(SP-TAT-apoptin),to test the hypothesis that it gains an additive bystander effect as an anti-cancer therapy. We used an artificial human secretory SP whose amino acid sequence and corresponding cDNA sequence were generated by the SP hidden Markov model. RESULTS:In human liver carcinoma HepG2 cells,SP-TAT-apoptin expression showed a diffuse pattern in the early phase after transfection. After 48 h,however,it translocated into the nuclear compartment and caused massive apoptotic cell death,as determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay and annexin-V binding assay. SP-TAT-apoptin did not,however,cause any cell death in non-malignant human umbilical vein endothelial cells(HUVECs). Most importantly,the conditioned medium from Chinese hamster ovary(CHO) cells transfected with SP-TAT-apoptin also induced significant cell deathin HepG2 cells,but not in HUVECs. CONCLUSION:The data demonstrated that SP-TAT-apoptin induces apoptosis only in malignant cells,and its secretory property might greatly increase its potency once it is delivered in vivo for cancer therapy.

Research advances of secretory proteins in malignant tumors

Secretory proteins in tumor tissues are important components of the tumor microenvironment.Secretory proteins act on tumor cells or stromal cells or mediate interactions between tumor cells and stromal cells,thereby affecting tumor progression and clinical treatment efficacy.In this paper,recent research advances in secretory proteins in malignant tumors are reviewed.

Structure and function of epididymal protein cysteine-rich secretory protein-1

Cysteine-rich secretory protein-1 （CRISP-1） is a glycoprotein secreted by the epididymal epithelium. It is a member of a large family of proteins characterized by two conserved domains and a set of 16 conserved cysteine residues. In mammals, CRISP-1 inhibits sperm-egg fusion and can suppress sperm capacitation. The molecular mechanism of action of the mammalian CRISP proteins remains unknown, but certain non-mammalian CRISP proteins can block ion channels. In the rat, CRISP-1 comprises two forms referred to as Proteins D and E. Recent work in our laboratory demonstrates that the D form of CRISP-1 associates transiently with the sperm surface, whereas the E form binds tightly. When the spermatozoa are washed, the E form of CRISP-1 persists on the sperm surface after all D form has dissociated. Cross-linking studies demonstrate different protein-protein interaction patterns for D and E, although no binding partners for either protein have yet been identified. Mass spectrometric analyses revealed a potential post-translational modification on the E form that is not present on the D form. This is the only discernable difference between Proteins D and E, and presumably is responsible for the difference in behavior of these two forms of rat CRISP- 1. These studies demonstrate that the more abundant D form interacts with spermatozoa transiently, possibly with a specific receptor on the sperm surface, consistent with a capacitation-suppressing function during sperm transit and storage in the epididymis, and also confirm a tightly bound population of the E form that could act in the female reproductive tract. （Asian J Androl 2007 July; 9： 508-514）

Quantitative Screening of Secretory Protein Genes in <i>Candidatus</i>Liberibacter Asiaticus

Huanglongbing (HLB) is the most destructive disease of citrus worldwide. The disease is caused by Candidatus Liberibacter spp., which is vectored by the psyllids Diaphorina citri Kuwayama and Trioza erytreae. Secretory proteins are important in bacterial pathogenesis and structure components. Some of them are expressed at a high level. To obtain the highly-expressed secretory protein genes (SPGs) for antiserum preparation, six candidate SPGs were chosen from Candidatus Liberibacter asiaticus by bioinformatic analysis and were further tested by qPCR and RT-qPCR methods, respectively. The result showed that two SPGs, 408 and pap (both are Flp pilus assembly protein genes), have relative high amounts of DNA and RNA transcripts of early HLB-infected green orange leaves. The 408 and pap genes were further constructed into the plant expression vector pCAMBIA1300 (GV1300: GFP) and expressed in tobacco leaf epidermal cells for subcellular localization analysis. The transient expression results indicated that the 408 protein is located in the nuclei and cytoplasm of tobacco leaf cells. However, the pap protein is located in the cytoplasm of tobacco leaf cells, which may help the pathogen invade into plant cells. This research is an important foundation for the preparation of the antiserum against Candidatus Liberibacter asiaticus and the early detection of HLB disease.

Distribution of secretory inhibitor of platelet microbicidal protein among anaerobic bacteria isolated from stool of children with diarrhea

AIM： To study the secretory inhibitor of platelet microbicidal protein （SIPMP） phenotypes of faecal anaerobic isolates from patients with diarrhea.METHODS： Faecal isolates of anaerobic bacteria （B. fragilis, n = 42; B. longum, n = 70; A. israelii, n = 21; E. lentum, n = 12） from children with diarrhea were tested. SIPMP production was tested by inhibition of platelet microbicidal protein （PMP） bioactivity against B. subtilis and was expressed as percentage of inhibition of PMP bactericidal activity.RESULTS： Among anaerobic isolates 80% of B. longum strains, 85.7% of A. israeli/strains, 50% of E. lentum strains and 92.86% of B. fragilis strains were SIPMP-positive. The isolated anaerobic organisms demonstrated SIPMP production at a mean level of 13.8% ± 0.7%, 14.7% ± 1.8%, 3.9% ± 0.9% （P 〈 0.05） and 26.8% ± 7.5% （P 〈 0.05） for bifidobacteria, A. israelii, E. lentum and B. fragilis, respectively.CONCLUSION： Data from the present study may have significant implications in understanding the pathogenesis of microecological disorders in the intestine, as well as for future improvement in the prevention and therapy of anaerobe-associated infections.

PnSCR82,a small cysteine-rich secretory protein of Phytophthora nicotianae,can enhance defense responses in plants

A number of plant pathogenic species of Phytophthora are known to produce different classes of secretory proteins during interactions with their hosts.Although several small cysteine-rich(SCR)secretory proteins,conserved in oomycete pathogens,have been identified in Phytophthora,their specific involvement in these interactions remains unknown.In this study,an SCR effector encoded by Pn SCR82 in P.nicotianae was identified and shown to have similarities to P.cactorum phytotoxic protein,Pc F(Phytophthora cactorum Fragaria).Agroinfection with potato virus X vector,Pn SCR82,was capable of inducing plant hypersensitive cell death in Nicotiana benthamiana and Solanum lycopersicum.Real-time PCR results indicated that transiently expressed Pn SCR82 in N.benthamiana leaves activated the jasmonate,salicylic acid and ethylene signaling pathways.Transient expression of Pn SCR82 enhanced plant resisitance to P.capsici.In summary,our results demonstrated that P.nicotianae Pn SCR82 elicits defensive responses in N.benthamiana and may potentially play a significant role in future crop protection programs.

Analysis of Diagnostic Efficiency of Excretory-Secretory Antigens for <i>Trichinella spiralis</i>and <i>Trichinella nativa</i>in ELISA

The comparative studies of diagnostic efficiency of excretory-secretory antigens of Trichinella spiralis and Trichinella nativa were performed using blood sera of rats from Wistar line experimentally infected with Arctic trichinellae. For animal infection and antigen preparation Trichinella from muscles of wild carnivorous mammals from Arctic regions of Russia were used. When antigen from T. nativa larvae was used to analyze titers of sera of rats experimentally infected with Arctic Trichinella, a significant increase in efficacy ofELISAwas detected. E.g., sera of rats infected with trichinellae from ringed seals retained in ELISA with T. nativa antigen values higher than diagnostic level at titers of 1:6400 - 1:12800, while titer of those same sera when using T. spiralis antigen was no higher than 1:200 - 1:400.

Correlation of serum mesothelin and human epididymis secretory protein 4 contents with cellular infiltrative growth in patients with ovarian cancer

Objective: To study the correlation of serum mesothelin and human epididymis secretory protein 4 contents with cellular infiltrative growth in patients with ovarian cancer. Methods:Patients with ovarian cancer who underwent surgical resection in West Coast Medical Center of the Affiliated Hospital of Qingdao University between June 2013 and December 2016 were selected as the ovarian cancer group of the study, and healthy women who received physical examination in China University of Petroleum (East China) Hospital during the same period were selected as the control group of the study. Serum was collected from two groups of subjects to detect serum mesothelin and human epididymis secretory protein 4 contents, and the ovarian cancer lesions and adjacent lesions were collected from ovarian cancer group to detect the expression of proliferation, apoptosis and invasion genes. Results: Serum mesothelin and human epididymis secretory protein 4 contents of ovarian cancer group were significantly higher than those of control group;FUNDC1, LSD1, TNFAIP8, CXCR4, β-catenin, CD44, PELP1, Slug, ZEB1 and Snail mRNA expression in ovarian cancer lesions were significantly higher than those in adjacent lesions and positively correlated with serum mesothelin and human epididymis secretory protein 4 contents while MFN2, PTEN, FN14 and E-cadherin mRNA expression were significantly lower than those in adjacent lesions and negatively correlated with serum mesothelin and human epididymis secretory protein 4 contents. Conclusion: Serum mesothelin and human epididymis secretory protein 4 contents abnormally increase in patients with ovarian cancer and are associated with the infiltrative growth of cancer cells.

冠心病患者血清VCAM-1、miR-145、Gal-3、SFRP5水平变化

目的探讨冠心病患者血清血管细胞黏附分子-1(VCAM-1)、miR-145、半乳糖凝集素-3(Gal-3)、分泌型卷曲蛋白5(SFRP5)水平变化及意义。方法选取冠心病患者80例为冠心病组,另收集健康志愿者40名为健康对照组。采集清晨空腹肘静脉血,酶联免疫吸附法测定血清VCAM-1、Gal-3、SFRP5浓度;RT-PCR检测血清miR-145表达水平。根据冠脉造影检查诊断的病变支数分为单支病变、双支病变、多支病变。收集两组受试者人口学特征及冠心病患者实验室指标;进行1 a随访,记录不良预后发生情况(病情加重再入院、死亡)。结果冠心病组患者血清VCAM-1、Gal-3水平明显高于健康对照组,miR-145、SFRP5水平明显低于健康对照组(均P<0.01)。急性心肌梗死组患者血清VCAM-1、Gal-3水平明显高于不稳定型心绞痛组、稳定型心绞痛组患者,血清miR-145、SFRP5水平明显低于不稳定型心绞痛组、稳定型心绞痛组患者(均P<0.05)。多支病变患者血清VCAM-1、Gal-3水平明显高于双支病变和单支病变患者,血清miR-145、SFRP5水平明显低于双支病变和单支病变患者(均P<0.05)。预后不良组患者血清VCAM-1、Gal-3水平明显高于预后良好组患者,miR-145、SFRP5水平明显低于预后良好组患者(均P<0.01)。VCAM-1、miR-145、Gal-3、SFRP5水平是冠心病患者预后的独立影响因素;ROC曲线分析显示,血清VCAM-1、miR-145、Gal-3、SFRP5水平联合检测对冠心病患者预后具有较高的预测价值(AUC=0.928)。结论冠心病患者血清VCAM-1、Gal-3水平高表达,miR-145、SFRP5水平低表达,且与冠心病分类、冠脉病变支数密切相关,联合检测对预后具有较高的预测价值。

血清SDC-1、HDAC6、CC16水平联合检测对慢性阻塞性肺疾病患者预后不良的预测价值

目的:探讨血清多配体蛋白聚糖1(SDC-1)、组蛋白去乙酰化酶6(HDAC6)、克拉拉细胞分泌蛋白16(CC16)水平联合检测对慢性阻塞性肺疾病(COPD)患者预后不良的预测价值。方法:选取2020年11月至2023年11月该院收治的147例COPD患者进行横断面研究,设为研究组;选取同期于该院体检的147名健康者设为对照组。比较两组、不同COPD病情程度患者、不同预后COPD患者血清SDC-1、HDAC6、CC16水平;采用受试者工作特征(ROC)曲线分析入院时血清SDC-1、HDAC6、CC16水平单项及联合检测预测COPD患者预后不良的价值。结果:研究组血清SDC-1、HDAC6水平均高于对照组,血清CC16水平低于对照组,差异有统计学意义(P<0.05)。重度COPD患者血清SDC-1、HDAC6水平高于中重度、中度、轻度患者,且中重度高于中度、轻度患者,中度高于轻度患者;重度COPD患者血清CC16水平低于中重度、中度、轻度患者,且中重度低于中度、轻度患者,中度低于轻度患者,差异均有统计学意义(P<0.05)。预后不良COPD患者入院时血清SDC-1、HDAC6水平高于预后良好患者,血清CC16水平低于预后良好患者,差异均有统计学意义(P<0.05)。ROC曲线分析结果显示,入院时血清SDC-1、HDAC6、CC16水平联合检测预测COPD患者预后不良的曲线下面积(AUC)为0.938,高于三者单项检测诊断(AUC=0.774、0.771、0.716)。结论:入院时血清SDC-1、HDAC6、CC16水平联合检测预测COPD患者预后不良的价值高于三者单项检测。

日本血吸虫成虫和虫卵排泄分泌抗原对Ⅰ型糖尿病模型小鼠的影响

目的观察日本血吸虫成虫排泄分泌抗原(adult-worm excretory-secretory protein,AES)和虫卵排泄分泌抗原(excretory-secretory antigen of eggs,ESA)对Ⅰ型糖尿病小鼠的影响,并初步探讨其作用机制。方法将24只雄性昆明鼠随机分为空白对照组、PBS组、AES组和ESA组,每组6只。空白对照组不作任何处理,后3组给予链脲佐菌素(streptozotocin,STZ)溶液注射制备Ⅰ型糖尿病模型,建模成功后分别用PBS、AES、ESA腹部皮下多点免疫,1周免疫1次,持续免疫4周。免疫干预1周后开始检测小鼠血糖,持续检测4周;免疫干预4周后采用ELISA法测定小鼠血清中IL-4和IFN-γ水平变化,采用HE染色法检测小鼠胰腺组织病理变化。结果与PBS组相比,AES组和ESA组小鼠血糖水平从免疫后第2周起出现下降趋势,且ESA组小鼠血糖水平低于AES组,免疫后第2、3、4周小鼠血糖水平组间差异有统计学意义(F=1214.00、1055.00、683.64,P均<0.05)。各组小鼠血清IL-4和IFN-γ水平组间差异有统计学意义(F=146.84、21.11,P均<0.05),ESA组小鼠血清中IL-4水平[(61.45±6.14)pg/mL]高于PBS组[(21.96±3.98)pg/mL]和AES组[(49.31±3.19)pg/mL],IFN-γ水平[(129.48±36.75)pg/mL]低于PBS组[(316.43±66.38)pg/mL]和AES组[(212.09±70.89)pg/mL],差异均有统计学意义(P均<0.05)。PBS组小鼠的胰岛萎缩,数量减少,空泡样变性;ESA组小鼠有轻微胰岛萎缩;AES组介于PBS组和ESA组之间。结论日本血吸虫ESA对Ⅰ型糖尿病小鼠有一定保护作用,其机制可能是ESA刺激机体导致Th1反应抑制和Th2反应增强,表现为IL-4升高和IFN-γ下降,在一定程度上减轻小鼠胰腺组织的病理损伤。

抗唾液腺蛋白1抗体联合抗腮腺分泌蛋白抗体对干燥综合征的诊断价值

目的:探讨抗唾液腺蛋白1(salivary protein 1, SP1)抗体联合抗腮腺分泌蛋白(parotid secretory protein, PSP)抗体在干燥综合征(Sj9gren’s syndrome, SS)诊断中的价值。方法:收集2020年1月至2022年12月就诊于河北医科大学第二医院风湿免疫科门诊及住院部的原发性SS(primary SS, pSS)患者60例,其他自身免疫病伴有口干和(或)眼干症状的患者30例为疾病对照组,体检中心的健康体检者30例为健康对照组,留取血清备用,同时记录一般资料、临床表现、实验室指标及其它辅助检查。采用2016年美国风湿病学会(American College of Rheumatology, ACR)/欧洲抗风湿病联盟(European League against Rheumatism, EULAR)干燥综合征分类标准作为pSS诊断金标准。采用化学发光免疫分析法检测免疫球蛋白G(immunoglobulin G,IgG)型抗SP1抗体和抗PSP抗体。用受试者工作特征(receiver operating characteristic, ROC)曲线评估抗SP1抗体和抗PSP抗体诊断pSS的准确性,并比较pSS组中抗SP1抗体和抗PSP抗体阳性患者与阴性患者的临床特征。采用t检验及Mann-Whitney U检验、方差分析、Kruskal-Wallis检验、卡方检验或Fisher确切概率法、Spearman相关分析进行统计学分析。结果:3组之间年龄(F=1.406,P=0.495)、性别(χ~2=2.105,P=0.349)差异无统计学意义。抗SP1抗体(H=16.73,P<0.001)和抗PSP抗体(H=26.09,P<0.001)在3组之间表达水平差异有统计学意义。组间比较发现,抗SP1抗体和抗PSP抗体表达水平在pSS和健康对照组之间差异均有统计学意义(P<0.001),抗PSP抗体表达水平在疾病对照组和健康对照组之间差异有统计学意义(P=0.009)。在pSS组、疾病对照组、健康对照组中,抗SP1抗体阳性率分别为58.33%vs. 40.00%vs.13.33%,差异有统计学意义(P<0.001);抗PSP抗体阳性率分别为75.00%vs. 56.17%vs.16.67%,差异有统计学意义(P<0.001)。抗SP1抗体、抗PSP抗体的曲线下面积分别为0.688(P<0.001)、0.720 (P<0.001),敏感性分别为58.33%(35/60)和75.00%(45/60),特异性分别为70.00%(42/60)和63.33%(38/60),阳性预测值分别为66.04%(35/53)和67.16%(45/67),阴性预测值分别为54.55%(42/77)和71.70%(38/53)。13例pSS患者中抗干燥综合征A (Sj9gren’s syndrome A, SSA,包括SSA52和SSA60)抗体和抗干燥综合征B (Sj9gren’s syndrome B, SSB)抗体均阴性,其中11例抗SP1抗体和抗PSP抗体均阳性,1例抗SP1抗体单独阳性,1例抗PSP抗体单独阳性。在pSS患者中,分别对抗SP1抗体和抗PSP抗体阳性与阴性组临床特征进行比较,抗SP1抗体阳性较阴性患者病程短(Z=-2.277,P=0.023);抗PSP抗体阳性较阴性患者比较:年龄相对较轻(t=2.598,P<0.05),类风湿因子(rheumatoid factor, RF)阳性率较高(P=0.002), IgG水平相对较高(t=3.806,P=0.003);对pSS患者抗SP1抗体和抗PSP抗体的相关性进行分析,发现二者之间存在明显的相关性(r=0.801,P<0.001)。结论:抗SP1抗体和抗PSP抗体在SS诊断中价值均较高,其中抗SP1抗体有助于pSS的早期诊断;两种抗体联合检测有助于抗SSA抗体和抗SSB抗体阴性pSS患者的早期诊断。

膝骨关节炎患者SFRP与玻璃酸钠治疗效果的关系

目的探讨膝骨关节炎(knee osteoarthritis,KOA)患者分泌型卷曲相关蛋白(secretory curl related protein,SFRP)水平与玻璃酸钠治疗效果的关系。方法选取2021年5月至2023年6月温州市中医院收治的KOA患者89例纳入观察组,所有患者均采用玻璃酸钠治疗,根据治疗效果分为显效/好转组和效果差组;同期选取67名健康体检者纳入对照组。采用酶联免疫吸附试验测定各组纳入者的SFRP1、SFRP2和SFRP5水平,并对KOA患者玻璃酸钠治疗效果进行多因素Logistic回归分析;绘制受试者操作特征曲线(receiver operating characteristic curve,ROC曲线),分析SFRP水平在KOA患者治疗效果中的评价效能。结果观察组患者的SFRP1、SFRP2及SFRP5水平低于对照组(P<0.05);89例KOA患者经玻璃酸钠治疗后,67例治疗效果良好(75.28%),显效/好转组患者的SFRP1、SFRP2及SFRP5水平高于效果差组(P<0.05);多因素分析结果表明,SFRP1、SFRP2、SFRP5水平及西安大略和麦克马斯特大学骨关节炎指数均为KOA患者治疗效果的影响因素(P<0.05);ROC曲线结果表明,SFRP1、SFRP2联合SFRP5对KOA患者治疗效果评价的敏感度和特异性高于单一指标(P<0.05)。结论SFRP1、SFRP2和SFRP5在KOA患者中呈低表达,其表达水平与患者玻璃酸钠治疗效果存在紧密联系,不同指标联合测定可提高预测的敏感度和特异性。

卵巢-附件影像报告和数据系统分类联合血清学检查对卵巢-附件肿瘤的诊断价值

目的探讨卵巢-附件影像报告和数据系统(O-RADS)分类联合血清糖类抗原125(CA125)及人附睾分泌蛋白4(HE4)对卵巢-附件肿瘤的诊断价值。方法回顾性分析安徽医科大学附属亳州医院(亳州市人民医院)收治并病理确诊的卵巢-附件肿瘤患者90例,以病理结果为金标准,分析超声O-RADS分类、血清CA125、HE4对卵巢-附件恶性肿瘤的检出率。运用受试者工作特征(ROC)曲线分析O-RADS分类、CA125、HE4及三者联合应用对卵巢-附件肿瘤的诊断效能。结果90例卵巢-附件肿瘤中,良性61例,恶性29例。O-RADS分类中2、3、4、5类对恶性肿瘤的检出率依次增高,与良性肿瘤比较,差异有统计学意义(P<0.001),血清CA125、HE4对恶性肿瘤的检出率分别为72.00%、70.37%,与良性肿瘤比较,差异有统计学意义(P均<0.001)。O-RADS分类、CA125、HE4及三者联合诊断卵巢-附件肿瘤的曲线下面积(AUC)分别为0.783、0.753、0.762、0.909,联合诊断AUC高于任一单独诊断技术(P均<0.05)。结论超声O-RADS分类联合血清CA125、HE4有助于提高卵巢-附件肿瘤的诊断效能。

金黄色葡萄球菌巨噬细胞胞内生存关键分泌蛋白筛选体系的构建

目的建立金黄色葡萄球菌(Staphylococcus aureus,S.aureus)巨噬细胞胞内感染状态下分泌蛋白高通量筛选体系,探索金葡菌胞内生存所需关键分泌蛋白。方法利用课题组建立的金葡菌分泌蛋白真核系统表达载体库,通过DNA转染表达技术,得到能够在细胞内表达金葡菌分泌蛋白的RAW264.7巨噬细胞阵列。金葡菌感染RAW264.7后,清除胞外菌,观察金葡菌胞内生存情况。最后通过构建相应分泌蛋白的过表达及敲除金葡菌验证筛选结果。结果通过探索RAW264.7质粒转染剂量、金葡菌感染复数(multiplicity of infection,MOI)以及感染时间,确立筛选体系的最优质粒转染剂量为1.0μg/孔、MOI为1.0、感染时间为4 h。筛选结果结合相应分泌蛋白过表达和敲除菌株验证,成功筛选出hypothetical protein、Serine protease E等分泌蛋白具有促进胞内金葡菌生存功能。结论成功构建金葡菌巨噬细胞胞内生存关键分泌蛋白筛选体系。