Candida glabrata is an opportunistic pathogen with increasing clinical significance due to its innate antifungal resistance. This study reveals the critical role of the phospholipid flippase Drs2 (P4-ATPase) in maintaining fungal pathogenicity and stress adaptation. The DRS2 deletion mutant exhibited impaired growth, heightened drug susceptibility, and defective stress responses, including compromised oxidative stress tolerance and autophagy. Importantly, the mutant showed reduced survival in macrophages while eliciting stronger proinflammatory responses, along with attenuated virulence in a Galleria mellonella infection model. Further analysis demonstrated diminished biofilm formation and impaired metabolic flexibility under nutrient-limited conditions. Through transcriptomic profiling, we identified Drs2 as a key regulator of MAPK signaling pathways, cell wall integrity, and oxidative stress responses. These findings collectively establish Drs2 as a central coordinator of membrane homeostasis that critically links cellular stress adaptation to virulence in C. glabrata. These findings identify Drs2 as a promising target for antifungal intervention and provide new insight into the biological functions of lipid flippases in fungal pathogens.

BACKGROUND:This study evaluated the performance of the simultaneous amplification and testing (SAT) assay for detecting common Candida species in vaginal specimens compared with culture (gold standard) at the Obstetrics and Gynecology Hospital of Fudan University from December, 2024 to January, 2025. METHODS:Speimens were analyzed by both Candida culture (with MALDI-TOF MS identification) and the SAT-Candida assay. Discordant results were further confirmed by real-time polymerase chain reaction (RT-PCR) and bidirectional sequencing. A comparative analysis (kappa coefficient) was conducted as well. We assessed the limit of detection, technical specificity, repeatability and the clinical diagnostic effectiveness, including sensitivity, specificity, diagnostic accuracy, positive predictive value (PPV), and negative predictive value (NPV) of SAT-Candida assay. RESULTS:In our study, among the 472 initially collected specimens, 5 were excluded because the presence of the Candida species were outside the detection spectrum of the SAT-Candida assay. The ages of the rest 467 cases ranged from 13 to 77 years old, and by comparison with SAT-Candida assay and Candida culture, 444 concordant results and 23 discordant results were discovered. After the reconfirmation, the SAT-Candida assay presented a overall sensitivity of 98.7%, specificity of 97.8%, PPV of 97.9%, NPV of 98.7%, and diagnostic accuracy of 98.3% in detecting Candida species. The kappa value between Candida culture and SAT-Candida assay in detecting Candida species was 0.91. CONCLUSIONS:This SAT-Candida assay is acute, highly sensitive, and specific, which can be applied as an optimal diagnostic tool for detecting and identifying common Candida species from vaginal samples of vulvovaginal candidiasis (VVC) suspected patients.

BackgroundInvasive candidiasis (IC) is an increasingly common, expensive, and potentially fatal infection. However, IC caused by multiple Candida species is rarely reported in China. Herein, we revealed a complex IC caused by multiple Candida species, comprising the rare C. norvegensis, C. albicans, C. glabrata, and C. tropicalis. The resistance mechanism of azole and echinocandin resistance were explored further.MethodsThe isolates were confirmed using internal transcribed spacer (ITS) sequencing. The resistance mechanisms were investigated using PCR-based sequencing, quantitative real-time reverse transcription PCR, and rhodamine 6G efflux quantification.ResultsAntifungal susceptibility testing showed this complex infection was associated with cross-resistance to azole and echinocandin drugs. For C. glabrata, the acquired echinocandin resistance was likely caused by a novel mutational pattern (1,3-beta-D-glucan synthase subunits FKS1-S629P and FKS2-W1497stop) while the acquired azole resistance in C. glabrata RJ05 was related to complex mechanisms including enhanced efflux activity, pleiotropic drug resistance 1 (PDR1) mutation, and increased expression of Candida drug resistance 1 (CDR1) and CDR2. Additionally, the azole resistance of C. tropicalis was caused by two lanosterol 14-alpha demethylase (ERG11) mutations: Y132F and S154F.ConclusionOur study revealed a case of clinically complex, multiple Candida invasive infections, further uncovering the resistance mechanisms to azoles and echinocandins. These findings provide valuable references for the diagnosis and treatment of invasive candidiasis (IC) in clinical practice.

IntroductionCandida glabrata has emerged as a fungal pathogen with high infection and mortality rates, and its primary virulence factors are related to adhesion and biofilm formation. These virulence factors in C.glabrata are primarily mediated by epithelial adhesins (Epas), most of which are encoded in subtelomeric regions and regulated by subtelomeric silencing mechanisms. The transcription factor Mss11, known for its regulatory role in adhesion, biofilm formation, and filamentous growth in Saccharomyces cerevisiae and Candida albicans, has also been implicated in the expression of EPA6, suggesting its potential influence on C.glabrata virulence. The present study aims to determine the regulatory role of Mss11 in the virulence of C. glabrata.MethodsIn this work, a Δmss11 null mutant and its complemented strain were constructed from a C.glabrata standard strain. The impact of the transcription factor Mss11 on the virulence of C.glabrata was investigated through a series of phenotypic experiments, including the microbial adhesion to hydrocarbons (MATH) test, adherence assay, biofilm assay, scanning electron microscopy and Galleria mellonella virulence assay. Furthermore, transcriptome sequencing, quantitative reverse transcription polymerase chain reaction (RT-qPCR), and chromatin immunoprecipitation sequencing (ChIP-seq) were employed to investigate the molecular mechanisms behind the regulation of Mss11.ResultsIn C.glabrata, the loss of MSS11 led to a significant reduction in several virulence factors including cell surface hydrophobicity, epithelial cell adhesion, and biofilm formation. These observations were consistent with the decreased virulence of the Δmss11 mutant observed in the Galleria mellonella infection model. Further exploration demonstrated that Mss11 modulates C. glabrata virulence by regulating EPA1 and EPA6 expression. It binds to the upstream regions of EPA1 and EPA6, as well as the promoter regions of the subtelomeric silencing-related genes SIR4, RIF1, and RAP1, indicating the dual regulatory role of Mss11.ConclusionMss11 plays a crucial role in C. glabrata adhesion and biofilm formation, and thus has a broad influence on virulence. This regulation is achieved by regulating the expression of EPA1 and EPA6 through both promoter-specific regulation and subtelomeric silencing.

BackgroundThe increasing incidence and high mortality rate of Candida glabrata infection in ICU patients is an important issue. Therefore, it is imperative to investigate the antifungal susceptibility profiles and epidemiological characteristics in local regions.MethodsHerein, antifungal susceptibility testing was conducted to determine the minimum inhibitory concentrations (MICs) of eight antifungal drugs. Multilocus sequence typing (MLST) was used to study the strain genotype, geographical distribution, and susceptibility to antifungal agents among C. glabrata isolates. The mechanism of echinocandin resistance was explored by sequencing the FKS1 and FKS2 genes (encoding 1,3-β-D-glucan synthases) of echinocandin-resistant C. glabrata strains. Moreover, we further investigated the clinical manifestations and the various risk factors of patients infected with C. glabrata in the ICU.ResultsWe selected 234 C. glabrata isolates from 234 patients in the ICU randomly for the follow-up study. Cross-resistance was found among the ICU C. glabrata isolates. Analysis using MLST showed that the genetic diversity among the C. glabrata isolates was low. Furthermore, sequence type showed no correlation with the antifungal resistance profiles, but was associated with geographical distribution. We also revealed novel mutations in FKS1 (S629P) and FKS2 (W1497stop) that mediated high-level echinocandin resistance (MIC >8 µg/mL). More than 14 days’ stay in ICU (P=0.007), Acute Physiology and Chronic Health Evaluation II (APACHE-II) score (P=0.024), prior antifungal exposure (P=0.039) and lung disease (P=0.036) were significantly associated with antifungal resistant/non-wild-type C. glabrata infection.ConclusionOur study shed light on the antifungal susceptibility, molecular epidemiology, and clinical risk factors of C. glabrata in the ICU of a Chinese Tertiary Hospital. Importantly, we revealed the molecular mechanism of echinocandin resistance. These results highlight the significance of continued surveillance in ICUs and provide data support for the treatment of C. glabrata in clinics.

目的 研究血清卷曲螺旋蛋白49(coiled-coil domain containing protein 49,CCDC49)对胃癌诊断的价值.方法 选取胃癌组患者和慢性胃炎组患者共288例,其中胃癌患者147例,慢性胃炎患者141例.采用双抗体夹心酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)测定血清CCDC49水平,统计分析血清蛋白CCDC49的诊断灵敏度、特异性,评估血清CCDC49在胃癌诊断中的临床价值.结果 胃癌组的CCDC49水平为0.251(0.124～1.103)ng/mL,慢性胃炎组的CCDC49水平为0.158(0.080～0.386)ng/mL.萎缩性胃炎组、浅表性胃炎组的CCDC49水平分别为0.164(0.094～0.553)ng/mL和0.133(0.034～0.237)ng/mL.胃癌组血清CCDC49水平高于慢性胃炎组,差异有统计学意义(P<0.05);胃癌组CCDC49水平与浅表性胃炎组差异有统计学意义(P<0.05);胃癌组CCDC49水平与萎缩性胃炎组差异无统计学意义(P>0.05);萎缩性胃炎组CCDC49水平与浅表性胃炎组差异无统计学意义(P>0.05).血清CCDC49的ROC曲线下面积为0.667,95％CI为0.555～0.779.当血清CCDC49水平为0.554ng/mL时,诊断胃癌的灵敏度和特异性分别为42.55％和82.93％.结论 血清CCDC49具有胃癌诊断的潜能,并优于血清CA19-9.

脂质组分在双分子膜中不对称分布是广泛存在于真核细胞中的现象,有助于膜系统的出芽和融合,从而促进胞吞胞吐、蛋白分选运输等生理过程.多种蛋白参与维持脂质不对称,其中P4型ATP酶是重要的脂质转运体.P4型ATP酶又称脂质翻转酶,主要介导氨基磷脂的转位.在酿酒酵母、新生隐球菌、白念珠菌和构巢曲霉中,多种P4型ATP酶亚基的缺失导致蛋白运输失常,对唑类药物的敏感性升高,细胞壁完整性受损,毒力减低,在巨噬细胞中生存能力减弱等表型,提示脂质翻转酶的重要性及其作为抗真菌靶点的潜力.

BACKGROUND:As highly-conserved types of lipid flippases among fungi, P4-ATPases play a significant role in various cellular processes. Cdc50 acts as the regulatory subunit of flippases, forming heterodimers with Drs2 to translocate aminophospholipids. Cdc50 homologs have been reported to be implicated in protein trafficking, drug susceptibility, and virulence in Saccharomyces cerevisiae, Candida albicans and Cryptococcus neoformans. It is likely that Cdc50 has an extensive influence on fungal cellular processes. The present study aimed to determine the function of Cdc50 in Candida glabrata by constructing a Δcdc50 null mutant and its complemented strain.RESULTS:In Candida glabrata, the loss of Cdc50 led to difficulty in yeast budding, probably caused by actin depolarization. The Δcdc50 mutant also showed hypersensitivity to azoles, caspofungin, and cell wall stressors. Further experiments indicated hyperactivation of the cell wall integrity pathway in the Δcdc50 mutant, which elevated the major cell wall contents. An increase in exposure of β-(1,3)-glucan and chitin on the cell surface was also observed through flow cytometry. Interestingly, we observed a decrease in the phagocytosis rate when the Δcdc50 mutant was co-incubated with THP-1 macrophages. The Δcdc50 mutant also exhibited weakened virulence in nematode survival tests.CONCLUSION:The results suggested that the lipid flippase subunit Cdc50 is implicated in yeast budding and cell wall integrity in C. glabrata, and thus have a broad influence on drug susceptibility and virulence. This work highlights the importance of lipid flippase, and offers potential targets for new drug research.

侵袭性真菌感染(invasive fungal infection, IFI)已经成为危害人类健康的严重威胁之一.尽管IFI的流行病学在过去20年中发生了变化,广谱抗真菌预防措施降低了传统危险因素患者的发病率,提高了患者的生存率,但由于一些稀有的真菌病原体以及部分耐药菌株出现了对现有药物的抗药性,因此新型抗真菌药物的研发迫在眉 睫 [1-2].本文列举了目前已有的新型抗真菌药物,总结其作用靶点机制,并对其研发进展进行综述.

结直肠癌(CRC)是第三大常见恶性肿瘤,也是全球癌症相关死亡的主要原因之一,每四分之一的CRC患者在诊断时都会伴随转移.因此,早发现、早诊断、早治疗是降低死亡率的关键.非侵袭性的生物标志物,因其安全、廉价、易于测量,是结直肠癌的早期筛查及辅助诊断的重要手段.本综述旨在讨论最近结直肠癌患者血液和粪便样本的新型诊断生物标志物研究进展.

引言 由于免疫功能低下的患者的数量增加和抗生素的广泛使用,真菌感染已成为医院中的重要问题,目前真菌引起的感染已影响到全世界数百万人口,根据统计调查,每年约有140万人死于真菌感染[1];最新的全球监视和多中心研究表明,超过90％的侵入性真菌感染(invasive fungal infec-tions,IFI)的病因是由念珠菌属(Candida spp.)感染引起的比[2-3].其中,光滑念珠菌(Candida gla-brata)占所有病例的近15％,且其占比有稳步上升的趋势,是仅次于白念珠菌(Candida albicans)的第二大常见的念珠菌病的病原体[6].

目的 对LiCA500全自动光激化学发光免疫分析系统及配套甲状腺项目检测试剂盒(化学发光法)进行临床分析性能验证.方法 依据国家卫生行业标准(WS/T492-2016)和美国临床实验室标准化协会(CLSI)EP系列文件的要求,对LiCA500全自动光激化学发光免疫分析系统检测抗甲状腺过氧化物酶抗体(thyroid peroxidase antibody,TPOAb)、抗甲状腺球蛋白抗体(thyroglobulin antibody,TGAb)和甲状腺球蛋白(thyroglobulin,TG)的精密度、准确度、线性范围和参考区间进行验证,并与厂商声明的性能标准进行比较.对40份血清进行检测,并与传统的放射免疫法检测结果进行比对.结果 LiCA500全自动光激化学发光免疫分析系统检测TPOAb、TGAb和TG的批内不精密度变异系数(CV)均≤8％,总不精密度均≤1/3 TEa;校准品检测均值与靶值的偏倚均≤1/2 TEa;线性回归方程斜率在0.992～1.010之间,R2均≥0.975;正常人血清样本的检测结果均在参考区间内;与放射免疫法检测结果进行线性回归分析,R2均≥0.95.结论 LiCA500全自动光激化学发光免疫分析仪检测TPOAb、TGAb和TG的分析性能符合要求,可用于临床上甲状腺疾病的诊断.

Background The Candida glabrata does not develop into a pathogenic hiphal form; however, it has become the second most common pathogen of fungal infections in humans, partly because of its adhesion ability and virulence. Objectives The present study aimed to determine whether Flo8, a transcription factor that plays an important role in the virulence and drug resistance in Candida albicans, has a similar role in C. glabrata. Methods We constructed FLO8 null strains of a C. glabrata standard strain and eight clinical strains from different sources, and a FLO8 complemented strain. Real-time quantitative PCR, biofilm formation assays, hydrophobicity tests, adhesion tests, Caenorhabditis elegans survival assay, and drug-susceptibility were then performed. Results Compared with the wild-type strains, the biofilm formation, hydrophobicity, adhesion, and virulence of the FLO8-deficient strains decreased, accompanied by decreased expression of EPA1, EPA6, and EPA7. On the other hand, it showed no changes in antifungal drug resistance, although the expression levels of CDR1, CDR2, and SNQ2 increased after FLO8 deletion. Conclusions These results indicated that Flo8 is involved in the adhesion and virulence of C. glabrata, with FLO8 deletion leading to decreased expression of EPA1, EPA6, and EPA7 and decreased biofilm formation, hydrophobicity, adhesion, and virulence.

目的:探讨多项指标联合检测在糖尿病肾病早期诊断中的临床意义.方法:随机选取2019年9月10日—19日本院诊治的150名2型糖尿病患者为观察对象,根据计算白蛋白(ACR)分为糖尿病正常蛋白尿(D M组)88名、早期糖尿病肾病组(EDN组)54名、临床糖尿病肾病组(CDN组)8名.同时选取同期50名体检健康者作为正常对照组,采用全自动生化仪测定血清胱抑素-C(CYS-C),肾小球滤过率(GFR)、尿转铁蛋白(TRF)、尿免疫球蛋白G(IGG)、尿α1-微球蛋白(α1-MG)及N-乙酰-β-D-氨基葡萄糖苷酶(NAG)的含量.结果:糖尿病各组的血清CYS-C,UTRF,UIGG,Uα1-MG尿NAG水平明显高于对照组,GFR低于对照组(P<0.05).CDN组血清CYS-C,UTRF,UIGG,Uα1-MG及尿NAG水平均高于DM组和EDN组,GFR水平低于DM组和EDN组(P<0.05);EDN组血清CYS-C,UTRF,UIGG,Uα1-MG及尿NAG水平高于DM组,GFR水平低于DM组(P<0.05).早期糖尿病肾病组联合检测阳性率显著高于各指标单项检测.结论:血CYS-C,GFR,尿TRF,UIGG,Uα1-MG及NAG联合检测能够迅速地判断早期糖尿病肾病,值得临床应用.

胃癌是全球4大最常见的恶性肿瘤之一,病死率居前7位,可见于胃窦、胃大小弯以及前后壁等区域.我国属胃癌高发地区,发病率及死亡率均占各类恶性肿瘤前列,但早期诊断率仅约10％.早期缺乏特异性症状与体征,晚期预后差,这对人类健康的影响与危害极大,形势相当严峻.早期根治性手术可以延长胃癌患者总生存期,如何提高胃癌的诊断效率,做到早发现早治疗,本文拟对血清学标志物在胃癌诊断中的临床价值做一综述.

目的 探讨血液透析后血清肾功能检测与血浆肾功能检测结果的相关性.方法 选择进行血液透析的患者200例,在血液透析后,分别同时抽血放入含有分离胶的普通试管(血清)及含有肝素锂抗凝剂的抗凝管(血浆),同批次进行检测,行相关性分析,以确定能否进行替代或换算.结果 血清CRE、URE、UA和血浆CRE、URE、UA检验结果比较差异无统计学意义(P>0.05).Pearson双侧相关性分析显示,血清CRE、URE、UA和血浆CRE、URE、UA均为极强正相关(P<0.0001).结论 血液透析后血清肾功能检测与血浆肾功能检测结果比较差异无统计学意义,两种检验方法结果具有显著的相关性,且两种方法的一致性好,临床实验室可以接受.

Objective: To investigate the performance of serum fibulin-1 (FBLN1) detection in diagnosis and prognosis prediction of colorectal cancer. Methods: A total of 81 patients with colorectal cancer(CRC group), 50 patients with benign colorectal polyps (polyps group) and 50 healthy individuals (healthy group) were enrolled. Serum levels of FBLN1 were detected by enzyme-linked immunosorbent assay(ELISA), CEA was also measured, and difference between groups was analyzed. Results: Similar to serum CEA, serum levels of FBLN1 were significantly higher in patients with colorectal cancer [(12.98±7.82) ng/mL] than in patients with benign colorectal polyps [(7.54±1.23) ng/mL] (P<0.05) and healthy individuals [(6.89±2.03) ng/mL] (P<0.05), respectively. No significant difference were found in FBLN1 level between patients with benign colorectal polyps and healthy individuals. The serum level of FBLN1 in patients with low differentiated CRC[ (22.23±18.51) ng/mL] was higher than that in patients with medium differentiated CRC [(11.65±13.28) ng/mL] (P<0.05) and patients with high differentiated CRC[(9.12±10.17) ng/mL] (P<0.05), respectively. The serum level of FBLN1 in CRC patients with lymph node metastasis[(15.26±6.98) ng/mL] was higher than that in patients with no lymph node metastasis [(8.58±4.27) ng/mL (P<0.05). The detection of FBLN1 for diagnosing CRC had a greater AUC(0.852) than CEA,with sensitivity of 82.7% and specificity of 90.0%. Conclusions: Serum FBLN1 can serve as a biomarker for diagnosis and prognosis prediction of CRC.

血清淀粉样蛋白A(SAA)属于急性时相反应蛋白,在感染急性期、炎症反应、组织损伤时迅速升高,在恢复期迅速下降.SAA与细菌或病毒感染、动脉粥样硬化、肿瘤、移植排斥反应、类风湿性关节炎、淀粉样病变等密切相关.文章主要就淀粉样蛋白(AA)超家族的结构、功能、检测方法及SAA在各种疾病中的作用机制及应用进展作一综述.

目的 探究血清三叶因子3(trefoil factor 3,TFF3)水平在结直肠癌诊断中的价值.方法 收集180例受试者(89例结直肠癌患者,44例结直肠息肉患者和47例健康对照者)血清,采用酶联免疫吸附法测定血清TFF3水平.将结直肠癌组血清TFF3水平与对照组作对比,绘制ROC曲线,评估血清TFF3作为肿瘤标志物对结直肠癌的诊断价值.结果 结直肠癌患者血清TFF3水平[12.35(9.72～13.92)ng/mL]显著高于结直肠息肉组(3.18±0.18ng/mL,P<0.05)和健康对照组(3.14±0.11ng/mL,P<0.05);与健康对照组相比,结直肠息肉组患者血清TFF3水平的升高差异无统计学意义(P>0.05).血清TFF3诊断结直肠癌的ROC曲线下面积为0.889,在cut-off值为4.6149ng/mL时,血清TFF3对结直肠癌的诊断灵敏度和特异性分别为73.7％和91.8％.淋巴结转移和分化程度低的结直肠癌患者,其血清TFF3水平显著升高(P<0.05).结论 本研究显示血清TFF3是一个能够有效诊断结直肠癌的血清标志物,具有一定的临床诊断价值.

目的 构建白念珠菌TFP1基因敲除株,并初步分析白念珠菌TFP1基因与钙调神经磷酸酶通路相关基因的关系.方法 通过融合PCR将白念珠菌TFP1基因上下游和营养缺陷筛选标记连接组成基因敲除组件,再运用醋酸锂转染法将一条基因敲除组件转入白念珠菌工程菌SN152,在营养缺陷平板上筛选出TFP1+/-菌株,再运用同样的方法敲除第2条等位基因.采用实时荧光定量PCR的方法检测TFP1-/-菌株、TFP1+/-菌株以及SN152菌株RTA2和CRZ1基因的表达量.结果 成功构建白念珠菌TFP1双等位基因敲除株,并且双等位基因敲除株中RTA2和CRZ1基因表达量均下降.结论 敲除白念珠菌TFP1基因影响钙调神经磷酸酶通路相关基因RTA2和CRZ1的表达.