BACKGROUND:Microsatellite instable-high (MSI-H) gastric cancer (GC) represents a distinct subgroup. However, controversy exists regarding the role of MSI in GCs, and the factors leading to internal prognostic differences among MSI-H GCs are rarely studied. METHODS:We identified 53 MSI-H cases from 941 consecutive GCs and conducted a detailed investigation of the clinical significance, clinicopathological correlations, and prognostic indicators of MSI-H GCs. RESULTS:Compared to MSI-low (MSI-L)/microsatellite stable (MSS) GCs, the MSI-H cohort was characterized by older age, female predominance, antral location, fewer lymph node (LN) metastases (H&E), and earlier tumor stage, but was also associated with larger tumor size, poor differentiation, and a high incidence of isolated tumor cell clusters (ITC) in negative LNs. ITC was then found to be correlated with tumor volume, Lauren subtype, pT stage, LN status (H&E), and lymphovascular invasion, with tumor size identified as an independent risk factor. Regarding prognosis, MSI-H GCs did not show longer survival time compared to MSI-L/MSS cases overall and in Stage Ⅲ-Ⅳ, but exhibited shorter survival time in Stage Ⅰ-Ⅱ. Moreover, in addition to age, pN stage, and distant metastasis, ITC and PD-L1 expression influenced survival in MSI-H GCs. ITC was confirmed as an independent unfavorable prognostic factor, while PD-L1 expression on interstitial immune cells independently predicted a favorable outcome. CONCLUSIONS:Our results suggest that MSI-H GC represents a peculiar clinicopathological entity with frequent occurrence of ITC in negative LNs. ITC and PD-L1 are crucial prognostic indicators for MSI-H patients.

Background Previously, only six cases of mixed neuroendocrine-non-neuroendocrine neoplasm (MiNENs) with squamous cell carcinoma (SCC) component have been described in the colorectum, and the molecular landscape of MiNENs is also poorly understood. Herein, we present a unique case in which the SCC developed as a component of a MiNEN in the rectum. Case presentation The patient was firstly diagnosed as rectal small cell neuroendocrine carcinoma (SCNEC) covered by tubulovillous adenoma, and then mixed SCNEC and SCC in the same site 6 months later. Representative samples from the three histologic subtypes were then sent for next-generation sequencing (NGS) separately. Multiple liver metastases occurred in the following month after the last surgery. The patient died of ketoacidosis 1 year after initial diagnosis of the tumor. Conclusion This is the first report of this exceedingly rare tumor type to include NGS of the 3 separate morphological entities. Our findings may expedite the understanding of combined tumors in the colorectum.

Objective To explore the clinical features and risk factors of preoperative anemia in patients with gastric cancer.Methods The clinical data of 218gastric cancer patients who received gastric cancer surgery in the Second Affiliated Hospital of Nanjing Medical University from January 2015 to December 2018 were retrospectively analyzed.Patients were divided into the anemia group(n = 84)and non-anemia group(n =134)based on hemoglobin levels.The gender,age,hypertension,diabetes,body mass index(BMI),carcinoembryonic antigen(CEA),cancer antigen 199(CA199),cancer antigen 724(CA724),tumor location,tumor size,clinical stage and lymph node metastasis were collected.The Logistic regression analysis was performed to analyze the risk factors of preoperative anemia in patients with gastric cancer.Results Among 218 patients with gastric cancer,the incidence of preoperative anemia was 38.53%,including 51 cases(60.7%)of mild anemia and 50 cases(59.5%)of normocytic anemia.There were no statistically signifi-cant differences in BMI,hypertension,diabetes,CEA,CA199,CA724,tumor location,and lymph node metastasis between the two groups(P>0.05),while there were statistically significant differences in the terms of gender,age,tumor size,and clinical stage be-tween the two groups(P<0.05).Multivariate Logistic regression analysis showed that gender(P = 0.002),age(P = 0.023),tumor size(P =0.001),and clinical stage(P =0.003)were independent risk factors for preoperative anemia in patients with gastric cancer.Conclusion Gender,age,tumor size and clinical stage are independent risk factors for preoperative anemia in patients with gastric canc-er,which is helpful for clinicians to take some intervention strategies for patients.

目的 探讨胃癌淋巴结转移风险预测模型的构建及评价.方法 回顾分析244例胃癌患者临床资料,根据是否发生淋巴结转移将患者分为转移组(n=171)和无转移组(n=73).采用Logistic回归分析胃癌淋巴结转移的危险因素,构建胃癌淋巴结转移的预测模型,采用受试者工作特征(ROC)曲线评价预测模型的价值.结果 Logistic回归分析发现,浸润深度、临床分期是胃癌淋巴结转移的独立危险因素(P<0.05).建立回归模型Logit(P)=-1.833+3.121×浸润深度(肌层)+2.799×浸润深度(穿透浆膜层)+5.468×临床分期,预测模型的ROC曲线下面积为0.958(95％CI:0.925～0.980,P<0.001),约登指数:0.815,cut-off值为0.715,敏感度和特异性分别为84.2％和97.3％.结论 浸润深度、临床分期是胃癌患者淋巴结转移的独立危险因素,该模型有助于临床预测胃癌患者是否发生淋巴结转移.

Mycobacterium tuberculosis 6-kDa early secretory antigenic target (ESAT-6) is a dominant target antigen for cell-mediated immunity in the early phase of tuberculosis. The fms-like tyrosine kinase 3 ligand (FL) that induces potent immune response has been used as an adjuvant in vaccine development. In this study, a new recombinant plasmid (pIRES-epitope-peptides-FL) encoding three T cell epitopes of ESAT-6 and FL was constructed, and the immunogenicity of the DNA vaccine was assessed in C57BL/6 mice immunized with the plasmid DNA vaccine. Additionally, a strategy of intramuscular injection with the DNA vaccine (prime) and intranasal administration of the epitope peptides (boost) was employed to induce higher immune reaction of the mice. The results showed that mice vaccinated with the recombinant plasmid DNA vaccine and boosted with the peptides not only increased the levels of Th1 cytokines (IFN-γ and IL-12), the number of IFN-γ(+) T cells and activities of cytotoxic T lymphocytes as well as IgG, but also enhanced protection against Mycobacterium tuberculosis challenge. In conclusion, these data indicate that the novel recombinant pIRES-epitope-peptides-FL plasmid is a useful DNA vaccine for preventing Mycobacterium tuberculosis infection.

肝样腺癌是一种发生于肝外的具有肝细胞癌样分化特点的高度侵袭性肿瘤，易转移，预后差。原发于胆管的肝样腺癌罕见，临床诊断中易被忽视。本文报道1例原发于胆总管并伴有肝转移的肝样腺癌，通过对其临床病理学特征及免疫组织化学特点进行分析并复习相关文献，总结其诊断及鉴别诊断要点。.

In addition to hepatoid adenocarcinoma (HAC), gastric adenocarcinoma with enteroblastic differentiation (GAED) and common adenocarcinoma (COM) could also show hepatoid differentiation, which presents a poor prognosis. To elucidate the histogenesis and development of gastric cancer with hepatoid differentiation, we identified 55 cases by histological morphology and a panel of markers, including α-fetoprotein (AFP), Glypican 3 (GPC3) and SALL4, then clinicopathological parameters, pathomorphological characteristics, mucin phenotypes, molecular features, Immunoscore and survival analysis were assessed. A mixture of three types (COM + GAED + HAC) was most commonly observed in the same case, and typical transitions between each histological subtype were frequently seen. Hyaline globule and pink amorphous substance were often present. HER2 was amplified in 21.8% of cases. All the tumors showed intestinal phenotype (69.1%) and mixed gastric/intestinal phenotype (30.9%) and were all defined to chromosomal instable (CIN)/genomically stable (GS) group. Considering that 83.6% cases presented TP53 gene mutation phenotype and 61.8% cases showed ≥10% aberrant E-cadherin expression, the precise molecule classification is ambiguous. Survival analysis showed that patients with high SALL4 expression, high preoperative serum AFP level, or low Immunoscore had a significantly poor overall survival (OS). Moreover, SALL4, HER2, and Immunoscore had an independent influence on OS. In conclusion, we suggest that the development of gastric adenocarcinoma with hepatoid differentiation might a continuously progressive profile: from intestinal-type COM adenocarcinoma to GAED and then HAC. CIN/GS subtypes might be where they belonged. SALL4, HER2, and Immunoscore may be potential therapeutic targets.

肝样腺癌属于特殊类型的胃癌,具有肝细胞癌样分化的组织学特点,通常伴血清AFP升高.胃肝样腺癌临床特征主要表现为老年男性多发,侵袭性强,多有脉管侵犯和肝脏转移,预后较差.该文旨在回顾性分析胃肝样腺癌的临床病理学特征、形态学表现、免疫表型和分子学特点,对胃肝样腺癌病理学发展进行综述.

目的 探讨非典型官颈腺细胞(AGC)在宫颈细胞学中的应用情况.方法 2012年1月～2019年8月选取南京医科大学第二附属医院病理科经液基薄层细胞检测检查的AGC病例34例,与其组织学结果进行对照并分析.结果 宫颈腺上皮异型增生7例(20.59％),宫颈腺癌2例(5.88％),子宫内膜腺癌4例(11.76％),高度鳞状上皮内病变累及腺体9例(26.47％),低分化鳞状细胞癌2例(5.88％),宫颈管内膜慢性炎7例(20.59％),子宫内膜息肉2例(5.88％),宫颈腺上皮微腺型增生1例(2.94％).根据病变类型,分为腺上皮病变组(13例)和鳞状上皮病变组(11例),两组年龄构成比较,差异有统计学意义(P＜0.05).结论 AGC在官颈组织学对照中,若为良性或者癌前病变,多数为官颈管腺上皮异形增生以及较大比例的需要鉴别的高度鳞状上皮内病变累及腺体;若为恶性,则多数为宫颈腺癌或子宫内膜腺癌及需要鉴别的低分化鳞癌.此外,不同病变类型可能与年龄有关系,40岁以上是腺上皮病变的高发年龄段.

Mesangioproliferative glomerulonephritis (MsPGN) is characterized by the proliferation of glomerular mesangial cells (GMCs) and accumulation of extracellular matrix (ECM), followed by glomerulosclerosis and renal failure of patients. Although our previous studies have demonstrated that sublytic C5b‐9 complex formed on the GMC membrane could trigger GMC proliferation and ECM expansion of rat Thy‐1 nephritis (Thy‐1N) as an animal model of MsPGN, their mechanisms are still not fully elucidated. In the present studies, we found that the levels of response gene to complement 32 (RGC‐32), myeloid zinc finger 1 (MZF1), phosphorylated extracellular signal‐regulated kinase 5 (phosphorylated ERK5, p‐ERK5), F‐box only protein 28 (FBXO28) and TNF receptor‐associated factor 6 (TRAF6) were all markedly up‐regulated both in the renal tissues of rats with Thy‐1N (in vivo) and in the GMCs upon sublytic C5b‐9 stimulation (in vitro). Further in vitro experiments revealed that up‐regulated FBXO28 and TRAF6 could form protein complex binding to ERK5 and enhance ERK5 K63‐ubiquitination and subsequent phosphorylation. Subsequently, ERK5 activation contributed to MZF1 expression and MZF1‐dependent RGC‐32 up‐regulation, finally resulting in GMC proliferative response. Furthermore, the MZF1‐binding element within RGC‐32 promoter and the functions of FBXO28 domains were identified. Additionally, knockdown of renal FBXO28, TRAF6, ERK5, MZF1 and RGC‐32 genes respectively markedly reduced GMC proliferation and ECM production in Thy‐1N rats. Together, these findings indicate that sublytic C5b‐9 induces GMC proliferative changes in rat Thy‐1N through ERK5/MZF1/RGC‐32 axis activated by the FBXO28‐TRAF6 complex, which might provide a new insight into MsPGN pathogenesis.

Sublytic C5b-9 formation on glomerular mesangial cells in rat Thy-1 nephritis (Thy-1N), a model of human mesangioproliferative glomerulonephritis, is accompanied by the production of proinflammatory cytokines, but the relationship between sublytic C5b-9 and cytokine synthesis and the underlying mechanism remains unclear. To explore the problems mentioned above, in this study, we first examined the levels of proinflammatory ILs (e.g., IL-23 and IL-36a) as well as transcription factor (KLF4) and coactivator (PCAF) in the renal tissues of Thy-1N rats and in the glomerular mesangial cell line (HBZY-1) stimulated by sublytic C5b-9. Then, we further determined the role of KLF4 and PCAF in sublytic C5b-9-induced IL-23 and IL-36a production as well as the related mechanism. Our results showed that the levels of KLF4, PCAF, IL-23, and IL-36a were obviously elevated. Mechanistic investigation revealed that sublytic C5b-9 stimulation could increase IL-23 and IL-36a synthesis through KLF4 and PCAF upregulation, and KLF4 and PCAF could form a complex, binding to the IL-23 or IL-36a promoter in a KLF4-dependent manner, causing gene transcription. Importantly, KLF4 acetylation by PCAF contributed to sublytic C5b-9-induced IL-23 and IL-36a transcription. Besides, the KLF4 binding regions on IL-23 or IL-36a promoters and the KLF4 lysine site acetylated by PCAF were identified. Furthermore, silencing renal KLF4 or PCAF gene could significantly inhibit IL-23 or IL-36a secretion and tissue damage of Thy-1N rats. Collectively, these findings implicate that the KLF4/PCAF interaction and KLF4 acetylation by PCAF play a pivotal role in the sublytic C5b-9-mediated IL-23 and IL-36a production of Thy-1N rats.

目的:研究转录因子KLF4调控sublytic C5b-9刺激肾小球系膜细胞（glomerular mesangial cell,GMC）诱导促炎因子白介素-23（IL-23）生成的作用。方法:构建KLF4的短发夹状小干扰RNA（shKLF4）及过表达质粒（pIRES2-KLF4）。将pIRES2-KLF4或shKLF4转染GMC后再行sublytic C5b-9刺激,用qPCR、Western blot和ELISA法检查沉默或过表达KLF4基因后对GMC产生IL-23的影响。此外,构建IL-23基因近端启动子全长质粒,行荧光素酶报告基因实验测定沉默或过表达KLF4基因后对IL-23启动子活性的影响。结果:（1）Sublytic C5b-9刺激GMC后能明显促进IL-23的生成,而沉默KLF4基因后,由sublytic C5b-9诱导GMC产生的IL-23明显减少,但过表达KLF4后IL-23的水平则显著增加。（2）Sublytic C5b-9刺激GMC能显著上调IL-23的启动子活性,而沉默KLF4基因后由sublytic C5b-9诱导的IL-23启动子活性明显降低,但过表达KLF4后又能显著升高IL-23的启动子活性。结论:Sublytic C5b-9刺激诱导IL-23的生成可通过其刺激上调转录因子KLF4的表达而实现。

BACKGROUND/AIMS:The activation of complement system and the formation of C5b-9 complex have been confirmed in the glomeruli of patients with mesangioproliferative glomerulonephritis (MsPGN). However, the role and mechanism of C5b-9-induced injury in glomerular mesangial cell (GMC) are poorly understood. Rat Thy-1N is an animal model for studying MsPGN. It has been revealed that the attack of C5b-9 to the GMC in rat Thy-1N is sublytic, and sublytic C5b-9 can cause GMC apoptosis, but the underlying mechanism is not fully elucidated. To explore the role and regulatory mechanism of C5b-9 in MsPGN lesion, we used rat Thy-1N model and first detected the change of microRNA (miRNA) profiles both in Thy-1N rat renal tissues (in vivo) and in the cultured GMCs with sublytic C5b-9 stimulation (in vitro). Then we determined the effect of miR-3546, which increased both in vivo and in vitro, on GMC apoptosis upon sublytic C5b-9 as well as the involved mechanism.METHODS:Rat Thy-1N model was established and GMCs were treated with sublytic C5b-9. The rat renal cortex and the stimulated GMCs were obtained for miRNA microarray detection. Subsequently, the increased miRNAs were verified by real-time PCR. Meanwhile, to ascertain the ability of some miRNAs to upregulate cleaved caspase 3 and induce GMC apoptosis, the corresponding miRNA mimics were transfected into GMCs, followed by western blotting (WB) and flow cytometry mesurement. Thereafter, the miR-3546-targeted gene (SOX4) was predicted using bioinformatics approaches, and SOX4 expression in Thy-1N tissues and in the GMCs upon sublytic C5b-9 stimulation or miR-3546 mimic/inhibitor transfection were detected using real-time PCR and WB. To prove that miR-3546 can affect SOX4 gene transcription and SOX4 can regulate survivin expression, dual luciferase reporter assay, real-time PCR, WB and chromatin immunoprecipitation (ChIP) assays were performed. Furthermore, the role of miR-3546/SOX4/survivin axis in the GMC apoptosis induced by sublytic C5b-9 was examined using WB and flow cytometry.RESULTS:Compared with normal renal tissues and untreated GMCs, there were 43 and 62 upregulated miRNAs (> 2-fold) in Thy-1N tissues and sublytic C5b-9-stimulated GMCs respectively. A total of 17 miRNAs were increased both in vivo and in vitro, 11 of which were validated by real-time PCR. Among them, miR-3546 could markedly promote GMC apoptosis and inhibit SOX4 or survivin expression in response to sublytic C5b-9, and either SOX4 or survivin overexpression markedly rescued the GMC apoptosis mediated by miR-3546 mimic. Additionally, SOX4 overexpression could reverse the survivin suppression by miR-3546 mimic, and SOX4 could bind to survivin promoter (-1,278 to -853 nt) and activate survivin gene transcription.CONCLUSION:MiR-3546/ SOX4/survivin axis has a promoting role in the GMC apoptosis triggered by sublytic C5b-9, and our findings may provide a new insight into the pathogenesis of rat Thy-1N and human MsPGN.

OBJECTIVE:By constructing the severe burns model in rat, we explored the effects of different doses of Ulinastatin (UTI) on protecting myocardium from oxidative stress and inflammatory reaction.MATERIALS AND METHODS:The severe burns model in rat was first constructed. Burned rats were intervened with different doses of UTI. Contents of cardiac troponin I (cTnI), Interleukin-1 (IL-1), Interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in rat serum and heart homogenate were detected by enzyme-linked immunosorbent assay (ELISA). Activities of SOD (superoxide dismutase), CAT (catalase), GSH-Px (glutathione peroxidase), and MDA (malondialdehyde) were detected by commercial kits. The inflammation and pathological changes in rat heart were observed by HE (Hematoxylin-Eosin) staining. Protein expressions of Cox-2, iNOS, NF-κB, Nrf2, and HO-1 in rat myocardium were detected by Western blot.RESULTS:Higher levels of cTnI, IL-1, IL-6, and TNF-α were found in model group than those of control group (p<0.05). Besides, decreased contents of cTnI, IL-1, IL-6, and TNF-α were observed in both UTI 50 ku/kg group and UTI 100 ku/kg group compared with those of model group (p<0.05). Decreased activities of SOD, CAT, and GSH-Px, as well as increased MDA level were observed in model group than those of control group (p<0.05). However, UTI treatment remarkably elevated SOD, CAT, and GSH-Px activities, whereas downregulated MDA level in burned rats (p<0.05). Abundant infiltration of inflammatory cells was found in the rat's myocardium of model group, which was alleviated in UTI group in a dose-dependent manner. Upregulated Cox-2, iNOS, and NF-κB, as well as downregulated Nrf2 and HO-1 were found in model group compared with those of control group (p<0.05). UTI pretreatment remarkably reversed the above-mentioned trends.CONCLUSIONS:Ulinastatin alleviates myocardial injury induced by severe burns. It exerts a protective role in myocardium via inhibiting oxidative stress and inflammatory response.

目的:鉴定亚溶解型C5b-9刺激大鼠肾小球系膜细胞(GMC)诱导Kruppel样因子4(KLF4)与P300/CBP相关因子(PCAF)相互结合以及结合部位.方法:以亚溶解型C5b-9刺激GMC,应用免疫沉淀(IP)和免疫印迹(IB)法检测KLF4与PCAF相互结合情况.另构建KLF4、PCAF过表达和KLF4、PCAF不同结构域的质粒,将质粒转染GMC,应用IP和IB鉴定KLF4与PCAF的结合部位.结果与结论:亚溶解型C5b-9刺激GMC后能够诱导KLF4与PCAF相互结合,KLF4结合于PCAF转录结合结构域,而PCAF则结合在KLF4的转录激活结构域.

Non-small cell lung cancer (NSCLC) is the most common type of lung cancer, and multiple evidence has confirmed that C5a production is elevated in NSCLC microenvironment. Although NSCLC cell proliferation induced by C5a has been reported, the involved mechanism has not been elucidated. In this study, we examined the proliferation-related genes (i.e., KLF5, GCN5, and GDF15) and C5a receptor (C5aR) expression in tumor tissues as well as C5a concentration in plasma of NSCLC patients, and then determined the roles of KLF5, GCN5, and GDF15 in C5a-triggered NSCLC cell proliferation and the related mechanism both in vitro and in vivo. Our results found that the expression of KLF5, GCN5, GDF15, C5aR, and C5a was significantly upregulated in NSCLC patients. Mechanistic exploration in vitro revealed that C5a could facilitate A549 cell proliferation through increasing KLF5, GCN5, and GDF15 expression. Besides, KLF5 and GCN5 could form a complex, binding to GDF15 promoter in a KLF5-dependent manner and leading to GDF15 gene transcription. More importantly, GCN5-mediated KLF5 acetylation contributing to GDF15 gene transcription and cell proliferation upon C5a stimulation, the region (−103 to +58 nt) of GDF15 promoter which KLF5 could bind to, and two new KLF5 lysine sites (K335 and K391) acetylated by GCN5 were identified for the first time. Furthermore, our experiment in vivo demonstrated that the growth of xenograft tumors in BALB/c nude mice was greatly suppressed by the silence of KLF5, GCN5, or GDF15. Collectively, these findings disclose that C5a-driven KLF5–GCN5–GDF15 axis had a critical role in NSCLC proliferation and might serve as targets for NSCLC therapy.

Objective:To construct rat p300 short hairpin RNA (shRNA) eukaryotic expression vector,and investigate the effect of silencing p300 gene on the apoptosis and activating transcription factor 3 (ATF3) acetylation in rat glomerular messangial cells (GMCs) stimulated by sublytic C5b-9.Methods:Three kinds of shRNAs targeting p300 gene were synthesized and cloned into eukaryotic expression vector pGCsi-U6/Neo/GFP/shRNA.The recombinant plasmids were transfected into cultured GMCs by NeonTM transfection system.P300 protein in the transfected cells was detected by Western blot to find out the optimal shRNA against p300 gene.The cell apoptosis was measured by flow cytometry and the acetylation of ATF3 was evaluated by Western blot combined with immunoprecipitation (IP) assays.Results:It was verified by nucleotide sequencing that the constructed p300 shRNAs were correct.Western blot assay showed that the p300 shRNA-2 was able to silence the target gene most effectively.Knockdown of p300 by shRNA in the GMCs reduced the number of GMC apoptosis as well as the level of ATF3 acetylation induced by sublytic C5b-9.Conclusion:The rat eukaryotic expression vector p300 shRNA was successfully constructed.It was preliminarily confirmed that p300 could promote GMC apoptosis triggered by sublytic C5b-9 through p300-regulated ATF3 acetylation.

The apoptosis of glomerular mesangial cells (GMCs) in the early phase of rat Thy-1 nephritis (Thy-1N), a model of human mesangioproliferative glomerulonephritis (MsPGN), is primarily triggered by sublytic C5b-9. However, the mechanism of GMC apoptosis induced by sublytic C5b-9 remains unclear. In this study, we demonstrate that expressions of TNFR1-associated death domain–containing protein (TRADD) and IFN regulatory factor–1 (IRF-1) were simultaneously upregulated in the renal tissue of Thy-1N rats (in vivo) and in GMCs under sublytic C5b-9 stimulation (in vitro). In vitro, TRADD was confirmed to be a downstream gene of IRF-1, because IRF-1 could bind to TRADD gene promoter to promote its transcription, leading to caspase 8 activation and GMC apoptosis. Increased phosphorylation of p38 MAPK was verified to contribute to IRF-1 and TRADD production and caspase 8 activation, as well as to GMC apoptosis induced by sublytic C5b-9. Furthermore, phosphorylation of MEK kinase 2 (MEKK2) mediated p38 MAPK activation. More importantly, three sites (Ser153/164/239) of MEKK2 phosphorylation were identified and demonstrated to be necessary for p38 MAPK activation. In addition, silencing of renal MEKK2, IRF-1, and TRADD genes or inhibition of p38 MAPK activation in vivo had obvious inhibitory effects on GMC apoptosis, secondary proliferation, and urinary protein secretion in rats with Thy-1N. Collectively, these findings indicate that the cascade axis of MEKK2–p38 MAPK–IRF-1–TRADD–caspase 8 may play an important role in GMC apoptosis following exposure to sublytic C5b-9 in rat Thy-1N.