Dual-state emission (DSE) is an emerging phenomenon wherein organic luminescent molecules display bright emissions in both molecularly isolated and packed states, addressing the challenge associated with the traditional paradigm of dyes with mono-state emission. This study presents the design and synthesis of two unsymmetrical triads, TPCA and TPCT, featuring a D-π-A-D′ electronic structure by integrating phenothiazines, triphenylamines, and cyanostilbene. Photophysical assessments reveal that both molecules serve as robust DSEgens, exhibiting strong emissions in both solution and solid phases. TPCA displays ΦTHF 53.2% and Φsolids 43.2%, while TPCT exhibits ΦTHF 49.6% and Φsolids 37.5%. However, due to differences in molecular conformation and packing, they diverge in solid-state emission wavelengths and mechanofluorochromic behavior. In the solid state, TPCA emits strong red fluorescence, contrasting with TPCT, which emits orange fluorescence. Furthermore, TPCA demonstrates significant mechanofluorochromism (MFC), shifting from yellow to yellow-red upon mechanical grinding, while TPCT exhibits negligible MFC owing to conformational distinctions. As robust and low-toxic bioimaging agents, both TPCA and TPCT prove highly effective for lipid-droplet imaging studies. This research contributes valuable insights to the evolving field of DSE materials, elucidating the promising applications and mechanisms governing their versatile emission behaviors. © 2024 Elsevier B.V.

Background: PD-1/PD-L1 inhibitors are approved treatments for patients with esophageal squamous cell carcinoma (ESCC). The present investigation aspired to explore the interrelation between molecular phenotype and PD-L1 expression in ESCC.Methods: PD-L1 testing and targeted next-generation sequencing (NGS) were performed on tumoral tissues from 139 ESCC patients. Tumor-infiltrating lymphocytes (TILs) were scrutinized using a tyramide signal amplification system combined with immunohistochemistry.Results: Among enrolled patients, 36.7% displayed high PD-L1 expression (combined positive score [CPS] >= 10). BRCA1 and NF1 gene mutations were significantly associated with high PD-L1 expression (p < .05) while TGF beta pathway alterations were linked to low PD-L1 expression (p = .02). High copy number instability (CNI) and copy number alterations (CNA) were correlated with low PD-L1 expression. Patients with CDKN2A deletion exhibited higher PD-L1 expression. Varying types of TILs were observed across different PD-L1 expression groups. The ratio of CD8(+)PD-L1(+) T cells and CD8(+)PD-1(+) T cells to CD8(+) T cells remained comparable in both tumoral and stromal regions, but the ratio of CD68(+)PD-L1(+) macrophages to CD68(+) macrophages was higher than the ratio of CD68(+)PD-1(+) macrophages to CD68(+) macrophages. CPS was significantly correlated with PD-L1(+) lymphocytes and CD68(+) macrophages in the tumoral region. CD8(+) T cell infiltration was positively correlated with PD-1(+) cells in both tumoral and stromal regions.Conclusion: In this study, we presented the prevalence rates of PD-L1 expression in Chinese ESCC patients. The association of genetic profiles with PD-L1 expression levels also provide the clue that genomic phenotype may interact with the immunologic phenotype in ESCC.

Cysticercosis pisiformis, a highly prevalent parasitic disease worldwide, causes significant economic losses in the rabbit breeding industry. Previous investigations have identified a novel microRNA, designated as novel-miR1, within the serum of rabbit infected with Cysticercus pisiformis. In the present study, we found that C. pisiformis-derived novel-miR1 was released into the rabbit serum via exosomes. Through computational analysis using TargetScan, miRanda, and PITA, a total of 634 target genes of novel-miR1 were predicted. To elucidate the functional role of novel-miR1, a dual-luciferase reporter assay was utilized and demonstrated that novel-miR1 targets rabbit Toll-like receptor 2 (TLR2). Rabbit peripheral blood lymphocytes (PBLCs) were transfected with novel-miR1 mimic and mimic NC, and the in vitro experiments confirmed that novel-miR1 suppressed the expression of pro-inflammatory cytokines such as TNF-alpha, IL-1 beta, and IL-6 through the nuclear factor kappa B (NF-kappa B) pathway. In vivo experiments demonstrated that novel-miR1 was significantly upregulated during the 1-3 months following infection with C. pisiformis in rabbits. Notably, this upregulation coincided with a downregulation of TLR2, P65, pP65, TNF-alpha, IL-1 beta, and IL-6 in PBLCs. Collectively, these results indicate that the novel-miR1 derived from C. pisiformis inhibited the rabbits' immune response by suppressing the NF-kappa B-mediated immune response. This immune modulation facilitates parasite invasion, survival, and establishment of a persistent infection.

Clonorchis sinensis is a zoonotic parasite associated with liver fibrosis and cholangiocarcinoma development. The role of toll-like receptors (TLRs) in C. sinensis infection has not yet been fully elucidated. Here, the TLR3 signaling pathway, cytokine expression and liver fibrosis were examined in C. sinensis-infected wildtype (WT) and TLR3(-/-) mice. Polyinosinic-polycytidylic acid (Poly (I:C)) was used to treat C. sinensis infections. The results showed that TLR3 deficiency caused severe clonorchiasis with increased parasite burden, exacerbated proinflammatory cytokine expression and liver lesions, promoted the TGF-beta 1/Smad2/3 pathway and myofibroblast activation, exacerbated liver fibrosis (compared to WT mice). Poly (I:C) intervention increased the body weight, decreased mouse mortality and parasite burden, reduced liver inflammation, and alleviated C. sinensis-induced liver fibrosis. Furthermore, C. sinensis extracellular vesicles (CsEVs) promote the production of IL-6, TNF in WT biliary epithelial cells (BECs) via p38/ERK pathway, compared with control group, while TLR3 deletion induced much higher levels of IL-6 and TNF in TLR3(-/-) BECs than that in WT BECs. Taken together, TLR3 inhibit IL-6 and TNF production via p38/ERK signaling pathway, a phenomenon that resulted in the alleviation of C. sinensis-induced liver fibrosis. Poly (I:C) is a potential treatment for clonorchiasis.

Author summaryClonorchis sinensis is one of the most prevalent and harmful food-borne parasitic diseases in Asian countries and is closely associated with cholangitis, cholecystitis, biliary fibrosis and cholangiocarcinoma. However, the pathogenesis of biliary fibrosis caused by C.sinensis has not been fully elucidated yet. In this study, we established a liver fibrosis model using TLR2-deficient mice and TLR2-normal mice to investigate the role and regulatory pathways of TLR2 in regulating biliary fibrosis induced by C. sinensis. A new role of TLR2 in aggravating C. sinensis-induced parasitic liver fibrosis was identified. C. sinensis ESPs activated TLR2-mediated AKT and p38 pathways to increase the production of IL-6 in mouse BECs in vitro. This result supported the finding that TLR2 triggered TGF-beta 1/Smad2/3 via promoting the production of IL-6 played crucial roles in activating myofibroblasts, which exacerbated liver fibrosis caused by C. sinensis. This study provides a useful mouse model to study the potential mechanisms of parasite-induced liver fibrosis, and explores the role of TLR2 deficiency in liver fibrosis, providing important reference information for understanding liver fibrosis caused by C. sinensis.Clonorchis sinensis is an important food-borne zoonotic parasite which has been linked to biliary fibrosis and cholangiocarcinoma. However, the details of the pathogenesis of C. sinensis were unclear. To explore the role and regulatory mechanism of toll-like receptor 2 (TLR2) in C. sinensis-induced biliary fibrosis, we established the C. sinensis-infected C57BL/6 mouse model with TLR2(-/-) and wild type (WT) mice. The mortality rate, liver lesions, TLR2 and TGF-beta 1 expression, phosphorylation of Smad2/3, AKT, p38, ERK and p65, and cytokine productions were analyzed. Furthermore, similar parameters were examined in mouse biliary epithelial cells (BECs) co-cultured with C. sinensis excretory/secretory proteins (ESPs). The results showed that TLR2 expression was enhanced significantly in C. sinensis-infected WT mice and mouse BECs. C. sinensis-infected TLR2(-/-) mice exhibited an increased weight and a decreased mortality rate; significantly alleviated liver lesions and biliary fibrosis, reduced numbers of myofibroblasts; decreased expression of TGF-beta 1 and phosphorylation level of AKT, p38 and Smad2/3; significantly decreased production of IL-6, TNF-alpha and IL-4, while increased production of IFN-gamma compared with C. sinensis-infected WT mice. Furthermore, C. sinensis ESPs could activate TLR2-mediated AKT and p38 pathways to increase the production of IL-6 in mouse BECs. In conclusion, these data indicate that C. sinensis infection activated TGF-beta 1-Smad2/3 through TLR2-mediated AKT and p38 pathways to promote IL-6 production, which resulted in myofibroblast activation and aggravating biliary fibrosis in mice.

为了深入研究猪带绦虫丝氨酸蛋白酶抑制分子（Ts-serpin-2）对THP-1细胞炎性细胞因子分泌的调节作用，本实验通过RTPCR从猪带绦虫成虫扩增获得Ts-serpin-2（Ts M＿000807300）的编码序列CDS，利用毕赤酵母表达Ts-serpin-2重组蛋白。利用发色底物特异性反应检测Ts-serpin-2蛋白对猪胰蛋白酶、胰弹性蛋白酶、牛α-糜蛋白酶、猪胃蛋白酶、木瓜酶、凝血酶和组织蛋白酶G的抑制效果。重组蛋白Ts-serpin-2处理THP-1细胞24 h，应用q RT-PCR和ELISA方法检测各炎性细胞因子差异表达水平。结果显示，真核酵母表达的重组蛋白Ts-serpin-2分子量约为48 kDa，具有蛋白酶抑制活性，对牛α-糜蛋白酶、猪胰弹性蛋白酶及猪胰蛋白酶的活性均有明显的抑制作用。重组蛋白Ts-serpin-2可抑制LPS活化的THP-1细胞促炎性细胞因子IL-1β、IL-6、IL-12α、IFN-γ和i NOS2m RNA的转录水平，促进IL-4刺激的THP-1细胞抗炎性细胞因子IL-10的表达。ELISA结果显示，Ts-serpin-2能够抑制巨噬细胞分泌致炎因子TNF-α、IL-6和IFN-γ，刺激IL-10的分泌，与q RT-PCR检测结果基本一致。上述研究结果表明猪带绦虫Ts-serpin-2可通过调节宿主巨噬细胞分泌的炎性因子影响虫体入侵过程中宿主的免疫应答。

目的探究旋毛虫感染早期如何诱导肠道病理变化及免疫调节相关细胞因子表达情况。方法通过苏木素伊红染色方法观察旋毛虫感染BALB/c鼠早期4个关键时间节点肠道病理学变化,采用电化学发光免疫分析法（Meso Scale Discovery,MSD）检测感染早期肠系膜淋巴结相关细胞因子表达情况。结果肠道病理学结果表明,肠粘膜增厚,嗜酸性粒细胞浸润,数量明显增多。MSD结果显示,感染后6 h肠系膜淋巴结Th1型细胞因子（IL-2）表达水平显著降低,其他细胞因子无显著变化;感染后3 d至6 d,Th1型细胞因子（IL-2、IFN-γ）和Th2型细胞因子（IL-4、IL-10）表达水平均显著升高,呈Th1/Th2混合型免疫应答。结论旋毛虫感染后6 h至6 d肠道炎症随感染时间延长加重,感染后6 h Th1型免疫应答受到抑制,随后旋毛虫通过诱导机体产生以Th2型为主的混合型免疫应答,实现旋毛虫在机体内的长期寄生。

近年来通过高通量测序技术发现了一类新型分节段黄病毒—荆门病毒属（Jingmenvirus）,主要成员包括Jingmen tick virus、Mogiana tick virus及Guaico Culex virus。该类病毒基因组分为4或5个节段,其中有2个节段来源于黄病毒的非结构蛋白,其他节段则表达病毒结构蛋白。该类病毒可感染蜱、蚊、牛、猴等动物,并且在中国、巴西、美国、乌干达地区都有分布。本研究在我国东北地区蜱咬伤病人血液中分离鉴定了一种新型分节段黄病毒-阿龙山病毒（Alongshan virus,ALSV）,并开展了人及动物流行病学研究。1.阿龙山病毒的分离鉴定:2017年4月采集哨点医院蜱咬伤病人血样进行宏病毒组学分析,发现1例病人血样中含有类似荆门蜱病毒（Jingmen tick virus,JMTV）的核酸序列,同源性57.4–74.5%。将血样接种Vero细胞,5–7天后可见轻微细胞病变;电镜负染观察,可见直径80–100 nm的带囊膜的近球形病毒粒子,切片观察可见胞质中有病毒颗粒。病毒基因全长为11350 bp,分为4个节段。由于病人发现于内蒙古阿龙山地区,因此命名为阿龙山病毒（ALSV）。2.阿龙山病毒基因组生物信息学分析:同源性分析发现ALSV与JMTV氨基酸同源性为23.7–80.9%;蛋白质组学分析ALSV存在类似黄病毒NS5和NS2b-NS3片段的NSP1和NSP2非结构蛋白,以及VP1囊膜蛋白、VP2核衣壳蛋白、VP3膜蛋白三个结构蛋白。进化分析发现ALSV与JMTV处于黄病毒属、瘟病毒属、肝炎病毒属之间,单独形成一个分枝,且分别处于不同分枝,表明ALSV属于分节段的荆门病毒属（Jingmenvirus）但是区别于JMTV的分节段黄病毒新种。3.阿龙山病毒在蜱咬伤病人流行病学分析:2017年4–8月采集的374份蜱咬伤住院病人血样中共检出86例ALSV阳性病例,患者主要集中在内蒙古东部及黑龙江大兴安岭地区,男性居多（73.3%）,多为野外工作者（97.7%）,有近期蜱叮咬史（95.3%）。病人发病潜伏期为3–7天,以头痛（80.2%）、发烧（77.9%）、疲乏（59.3%）为主要症状,住院10-14天可痊愈。4.阿龙山病毒在动物的流行病学分析:采集内蒙古东部地区牛羊血清各240份,检测发现牛羊ALSV ELISA抗体阳性率为4.6–9.2%;中和抗体阳性率为1.7–4.2%,说明牛羊存在ALSV的既往感染。RT-qPCR检测牛羊ALSV阳性率为26.3–27.5%,表明牛羊存在ALSV近期感染,可能为ALSV的储存宿主。在黑龙江和内蒙古采集蜱1105只,通过巢式PCR在全沟硬蜱中检出ALSV,黑龙江感染率3.7%,内蒙古6.5%,表明全沟硬蜱可能为ALSV的传播媒介。遗传进化分析显示牛、羊、蜱及人源ALSV同源性在98%以上,且位于进化树同一分枝,表明各宿主源ALSV在进化上密切相关。本研究发现了一种新型分节段黄病毒—阿龙山病毒,是世界上发现的第一个感染人的分节段黄病毒,对于蜱传疾病的防控具有重要指导意义,同时也为蜱传病研究提供了新的方向。

The objective of this study is to detect the number of different subsets of TFH and B cells in renal transplant recipients (RTR) with antibody-mediated acute rejection (AMR), acute rejection (AR), chronic rejection (CR), or transplant stable (TS). The present study was a prospective study. The numbers of ICOS +, PD-1+ and IL-21+ TFH, CD86+, CD38+, CD27+, and IgD- B cells in 21 patients with end-stage renal disease (ESRD) and post-transplant times were measured by flow cytometry. The level of serum IL-21 was detected by ELISA. The numbers of circulating CD4+CXCR5+, CD4+CXCR5+ICOS+, CD4+CXCR5+PD-1+, CD4+CXCR5+IL-21+ TFH, CD19+CD86+, and CD19 +CD86+CD38+ B cells as well as the level of serum IL-21 in the AMR, AR, and CR groups at post-transplantation were significantly higher than those at pre-transplantation. In contrast, the number of circulating CD19+CD27+IgD B cells was significantly increased in the TS groups in respect to the other groups. Moreover, the numbers of circulating CD4+CXCR5+IL-21+ TFH cells, CD19+CD86+CD38+ B cells as well as the level of serum IL-21 were positive related to the level of serum Cr while showing negative correlated with the values of eGFR in the AMR groups at post-transplantation for 4 and 12 weeks. Circulating TFH cells may be a biomarker in RTR with AMR, which can promote the differentiation of B cells into plasma cells by activating B cells, thereby promoting disease progression.

Minks and brown rats are reservoir hosts for many endoparasites including those of the genus Trichinella, a group of parasite nematodes with a worldwide distribution. However, little is known about the prevalence of Trichinella sp. infection in the American mink (Neovison vison) and rats (Rattus norvegicus) in China. Therefore, we aimed to examine the prevalence of Trichinella sp. infection in farmed minks in Weihai city, Shandong province, China and infer the possible route for Trichinella transmission to farmed American minks. In total, 289 muscle samples from minks and 102 carcasses of rats were collected from Weihai City. The appearance of Trichinella sp. was examined using the pooled artificial HCl-pepsin digestion method. The results showed that muscle larvae were detected in 20 of 289 minks (6.92%) and 2 of 102 synanthropic rats (1.96%). The larval density of Trichinella sp. in mink samples ranged from 0.025 to 0.815 larvae per gram (lpg), while the average larval burden in rats was 0.17 lpg. The isolates derived from minks and rats were identified at the species level using multiplex polymerase chain reaction (PCR), which revealed that the size of the two PCR products matched that of T. spiralis at 173 bp. Furthermore, sequence analysis showed 100% identity of the 5S rDNA inter-gene spacer regions of the two isolates to that of T. spiralis. This study presents a novel report of T. spiralis-mediated infection in minks and synanthropic rats in China. We highlight the vulnerability of farmed minks to Trichinella infection through exposure to synanthropic rats, which may raise a public health concern of potential zoonotic risks for domestic animals.