Biliary hemorrhage caused by a malignant small round cell tumor in the common bile duct: A case report.

BACKGROUND: Malignant small round cell tumor (MSRCT) metastasis to the common bile duct associated with recurrent biliary hemorrhage is extremely rare. Thus far, there have been no reports of metastatic small round cell tumors of the common bile duct. CASE SUMMARY: Herein, we report the case of a 77-year-old female patient with an MSRCT in the common bile duct. The patient was admitted to hospital due to gastrointestinal hemorrhage and abdominal pain. We found a neoplasm in the common bile duct with active bleeding through a spyglass. We performed biopsy through the spyglass and placed a metal stent to stop bleeding. The pathological result suggested that it was an MSRCT metastasized from the back to the common bile duct. Later, we found using fluorescence in situ hybridization that the SS18 gene break test was negative, ruling out the diagnosis of synovial sarcoma. CONCLUSION: MSRCT is a group of tumors with similar cell morphology and diffuse histological structure. Complete tumor resection results in improved survival in patients with MSRCT. Roux-en-Y cholangiojejunostomy was performed. After excision of the common bile duct tumor, the patient felt that the abdominal pain improved and hemorrhage disappeared. The patient underwent routine fecal examination one month after surgery, indicating a negative fecal occult blood test. On May 22, 2023, the patient was reexamined by abdominal computed tomography, and no abdominal space occupying lesions or abdominal lymphadenopathy was found.

Sec1 regulates intestinal mucosal immunity in a mouse model of inflammatory bowel disease.

Inflammatory bowel disease (IBD) is a common immune-mediated condition with its molecular pathogenesis remaining to be fully elucidated. This study aimed to deepen our understanding of the role of FUT2 in human IBD, by studying a new surrogate gene Sec1, a neighboring gene of Fut2 and Fut1 that co-encodes the α 1,2 fucosyltransferase in mice. CRISPR/Cas9 was used to prepare Sec1 knockout (Sec1-/-) mice. IBD was induced in mice using 3% w/v dextran sulphate sodium. Small interfering RNA (siRNA) was employed to silence Sec1 in murine colon cancer cell lines CT26.WT and CMT93. IBD-related symptoms, colonic immune responses, proliferation and apoptosis of colon epithelial cells were assessed respectively to determine the role of Sec1 in mouse IBD. Impact of Sec1 on the expression of death receptor 5 (DR5) and other apoptosis-associated proteins were determined. Sec1 knockout was found to be associated with deterioration of IBD in mice and elevated immune responses in the colonic mucosa. Silencing Sec1 in CT26.WT and CMT93 cells led to greater secretion of inflammatory cytokines IL-1ß, IL-6 and TNF-α. Cell counting kit 8 (CCK8) assay, flow cytometry and TUNEL detection suggested that Sec1 expression promoted the proliferation of colon epithelial cells, inhibited cell apoptosis, reduced cell arrest in G0/G1 phase and facilitated repair of inflammatory injury. Over-expression of DR5 and several apoptosis-related effector proteins was noticed in Sec1-/- mice and Sec1-silenced CT26.WT and CMT93 cells, supporting a suppressive role of Sec1 in cell apoptosis. Our results depicted important regulatory roles of Sec1 in mouse IBD, further reflecting the importance of FUT2 in the pathogenesis of human IBD.

Sodium butyrate exerts antioxidant stress effects and attenuates Aß25-35-induced cytotoxicity in PC12 cells.

Alzheimer's disease (AD), a common neurodegenerative disease, is characterised by the deposition of amyloid-ß (Aß) plaques and neurofibrillary tangles. An increasing number of studies have demonstrated that Aß causes neuronal damage and mitochondrial dysfunction. Herein, we evaluated the neuroprotective effect of sodium butyrate (NaB) against Aß induced neurotoxicity in PC12 cells. The results revealed that 3 mM of NaB promoted the expression of angiotensin-converting enzyme and brain-derived neurotrophic factor, which exert a neuroprotective effect by activating G protein-coupled receptors. Moreover, NaB could significantly improve mitochondrial dysfunction caused by Aß. In conclusion, NaB protected PC12 cells from Aß-induced cell damage, highlighting the potential of NaB in AD treatment.

Influence of TRAIL Deficiency on Th17 Cells and Colonic Microbiota in Experimental Colitis Mouse Model.

BACKGROUND: The abnormalities of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) are implicated in various autoimmune disorders and tumors. This study investigated the influence of TRAIL deficiency on Th17 cells and colonic microbiota in experimental colitis mouse model. METHODS: Mice were randomly divided into 4 groups: wild-type, TRAIL gene knock-out (TRAIL-/-), wild-type colitis and TRAIL-/- colitis groups. Colitis was induced by oral administration of 3.5% dextran sulfate sodium (DSS) for 7 consecutive days. Mice were given scores for disease severity both clinically and histopathologically. Th17 cells in peripheral blood and mesenteric lymph nodes (MLNs) were assessed using flow cytometry. The expression levels of Th17 cell markers IL-17A and ROR-γt were evaluated by quantitative real-time polymerase chain reaction. The colonic samples were also analyzed for microbiota profile by 16s-rDNA gene sequencing on variable V4 region. RESULTS: Compared with wild-type counterparts, TRAIL-/- mice developed more severe colitis after DSS treatment. Colitis TRAIL-/- mice had increased proportion of Th17 cells and elevated mRNA expression levels of IL-17A and ROR-γt in peripheral blood and MLNs compared with colitis wild-type mice. In contrast to colitis wild-type mice, the composition of colonic microbiota was shifted in colitis TRAIL-/- mice, and was characterized by increased alpha diversity, increased TM7, deferribacteres and tenericutes, and decreased proteobacteria at the phylum level. CONCLUSIONS: These findings suggested that TRAIL deficiency not only aggravated DSS-induced colitis, but also led to enhanced Th17 cell response and altered colonic microbiota composition.

Combined toxic effects of CBNPs and Pb on rat alveolar macrophage apoptosis and autophagy flux.

Carbon black (CB) and heavy metals are the main components of Particulate Matter (PM). Although the individual toxicities of CB and heavy metals have been extensively studied, the combined toxicity is much less understood. In this study, we choose the nano carbon black (CBNPs) and Pb2+ to simulate fine particles in the atmosphere and study the combined toxic effect on rat alveolar macrophages. The data showed that CBNPs could adsorb Pb2+ to form CBNPs-Pb2+ complex and displayed an altered physical properties by particle characterization. CBNPs-Pb2+ synergistically induced rat alveolar macrophages apoptosis and blocked autophagy flux compared with CBNPs and Pb2+ individually. Consistent with this, CBNPs-Pb2+ could impair the mitochondrial membrane potential (MMP), activate apoptotic signaling pathways, inhibit lysosomal function.

Low-Dose Radiation Promotes the Proliferation and Migration of AGE-Treated Endothelial Progenitor Cells Derived from Bone Marrow via Activating SDF-1/CXCR4/ERK Signaling Pathway.

Low-dose radiation (LDR) has been confirmed to mobilize bone marrow-derived endothelial progenitor cells (EPCs) and promote diabetic wound healing. But it is unclear whether LDR acts directly on EPCs and promotes their proliferation and migration. Given the key role of advanced glycosylation end products (AGE) in the pathogenesis of diabetes, we used AGE to induce EPC damage. We then investigated the effect of LDR on the proliferation and migration of AGE-treated EPCs and explored the underlying mechanisms. EPCs cultured in vitro were treated with different concentrations of AGE, and the cells were then exposed to different low doses and treated with a specific antagonist for CXCR4, AMD3100 (1 lmol/l). The proliferation and migration abilities of EPCs were detected using the CCK-8 and wound healing assays, respectively. The mRNA and protein expression of SDF-1 and CXCR4 in AGE-treated EPCs were measured using qPCR and Western blot analysis, respectively. The expressions of ERK and phosphorylated ERK (pERK) were detected using Western blot analysis. The results showed that 200 mg/l and 400 mg/l AGE had an inhibitory effect on the proliferation of EPCs, and this inhibitory effect was exerted in a dose- and time-dependent manner. AGE significantly reduced the migration ability of EPCs cultured in vitro. After the cells received either 50 or 75 mGy low-dose irradiation, the proliferation of EPCs and AGE-treated EPCs was clearly increased; in addition, LDR also enhanced cell migration ability, but this enhancement was counteracted by AMD3100. Results from qPCR and Western blot analysis showed that LDR increased the mRNA and protein expression of SDF-1/ CXCR4. LDR also upregulated pERK expression in EPCs and AGE-treated EPCs, but LDR-induced upregulation of pERK expression was inhibited by AMD3100. These findings indicate that LDR can directly activate the SDF-1/CXCR4 biological axis and downstream ERK signaling pathway, and promote the proliferation and migration abilities of EPCs by increasing the expression of SDF-1, CXCR4 and pERK in EPCs.

Epigallocatechin-3-gallate protects against 1,3-dichloro-2-propanol-induced lipid accumulation in C57BL/6J mice.

AIMS: Chloropropanol is a contaminant produced during food processing, and 1,3-dichloro-2-propanol (1,3-DCP) is one of the most-studied and most common chloropropanol-related food contaminants. Epigallocatechin-3-gallate (EGCG) is the most abundant ester catechin in tea polyphenols. We studied the potential therapeutic effect of EGCG on 1,3-DCP-induced lipid accumulation in the liver of mice, and determined the related regulatory mechanisms. MATERIALS AND METHODS: The effects of EGCG were investigated in 6-8-week-old adult male C57BL/6J mice that were given 1,3-DCP (1 mg/kg bw/day; i.g.) for 6 weeks. EGCG (10, 31.6 and 100 mg/kg bw/day i.g.) was administered daily in the 1,3-DCP-treated mice for 10 days. Total cholesterol (TC) and triglyceride (TG) were measured in serum and liver. For histological examination, HE staining and oil red O experiments were performed. Western blot and quantitative RT-PCR were subsequently used to study the molecular mechanisms. KEY FINDINGS: Increasing concentrations of EGCG significantly lowered TC and TG levels compared with those of the model group. Furthermore, EGCG dramatically increased expression of cAMP, P-PKA and P-CREBP, -AMPKα (Tr172), LKB1, P-ACC (Ser79) and lowered expression of CD36, SREBP-2, HMGCR, SREBP-1, GPAT in 1,3-DCP-treated mice livers. Quantitative RT-PCR experiments showed that EGCG regulated gene transcription of AMPK, SREBF-2, HMGCR and SREBP-1c. SIGNIFICANCE: These data suggested that EGCG intervention restored 1,3-DCP-altered protein levels and reduced hepatic lipid levels to normal. The mechanism was mediated by the AMPK and PKA pathways. EGCG may be developed as a candidate natural agent for the treatment of 1,3-DCP-induced lipid accumulation.

Urinary kidney injury molecule­1 as an early diagnostic biomarker of obstructive acute kidney injury and development of a rapid detection method.

The aim of the present study was to investigate whether urinary kidney injury molecule­1 (KIM­1) presents a suitable early diagnostic biomarker of obstructive nephropathy­induced acute kidney injury (AKI), and to develop a rapid detection method for urinary KIM­1. Obstructive AKI was induced in an experimental rat model by a unilateral ureteral obstruction (UUO) operation. Macro­ and micromorphological kidney alterations were determined by visual observation and hematoxylin and eosin (HE) staining, respectively. Kidney functions were evaluated by detecting urea nitrogen and creatinine levels in rat urine and blood. Urinary KIM­1 levels were measured using an enzyme­linked immunosorbent assay, and the protein expression levels of KIM­1, α­smooth muscle actin (α­SMA) and vimentin in kidney tissues were detected using immunohistochemical assays. In order to measure KIM­1 levels, colloidal gold immunochromatographic strips were developed based on the colloidal gold immunochromatographic assay. The results indicated that KIM­1 levels were significantly higher in the UUO group when compared with the Sham group. KIM­1 levels in the urine and kidney tissues exhibited a time­dependent increase, together with increasing obstructive AKI in the UUO group. In addition, KIM­1 levels were demonstrated to be a more sensitive biomarker of early obstructive AKI, when compared with α­SMA and vimentin. A colloidal gold­based immunochromatographic strip was developed, whereby the detection of urinary KIM­1 could be completed within 5­10 min. In conclusion, results of the present study demonstrated that urinary KIM­1 may be a valuable biomarker for the early diagnosis of obstructive AKI, and the use of a colloidal gold immunochromatographic strip may be a promising method for the rapid detection of urinary KIM­1.

Downregulation of microRNA-637 Increases Risk of Hypoxia-Induced Pulmonary Hypertension by Modulating Expression of Cyclin Dependent Kinase 6 (CDK6) in Pulmonary Smooth Muscle Cells.

BACKGROUND The objective of this study was to investigate the molecular mechanism by which miR-637 interferes with the expression of CDK6, which contributes to the development of pulmonary hypertension (PH) with chronic obstructive pulmonary disease (COPD). MATERIAL AND METHODS We used an online miRNA database to identify CDK6 as a virtual target of miR-637, and validated the hypothesis using luciferase assay. Furthermore, we transfected SMCs with miR-637 mimics and inhibitor, and expression of CDK6 was determined using Western blot and real-time PCR. RESULTS In this study, we identified CDK6 as a target of miR-637 in smooth muscle cells (SMCs), and determined the expression of miR-637 in SMCs from PH patients with COPD and normal controls. We also identified the exact miR-637 binding site in the 3'UTR of CDK6 by using a luciferase reporter system. The mRNA and protein expression levels of CDK6 in SMCs from PH patients with COPD were clearly upregulated compared with the normal controls. Cells exposed to hypoxia also showed notably increased CKD6 mRNA and protein expression levels, and when treated with miR-637 or CDK6 siRNA, this increase in CKD6 expression was clearly attenuated. Additionally, cell viability and cell cycle analysis showed that hypoxia markedly increased viability of SMCs by causing an accumulation in S phase, which was relieved by the introduction of miR-637 or CDK6 siRNA. CONCLUSIONS Our study proved that the CDK6 gene is a target of miR-637, and demonstrated the regulatory association between miR-637 and CDK6, suggesting a possible therapeutic target for PH, especially in patients with COPD.

Knockdown of toll-like receptor 4 inhibits human NSCLC cancer cell growth and inflammatory cytokine secretion in vitro and in vivo.

Toll-like receptor 4 (TLR4)-mediated signaling has been implicated in tumor cell invasion, survival and metastasis in several types of cancers. However, the expression of TLR4 in patients with non-small cell lung cancer (NSCLC) and its biological function in the development and progression of NSCLC have not been elucidated to date. Here, we sought to characterize the expression of TLR4 in patients with NSCLC and to investigate the biological roles of TLR4 in lung metastasis, cell invasion and survival. In this study, we found that TLR4 expression was elevated in most patients with NSCLC, and its expression levels correlated with key pathological characteristics, including tumor differentiation, stage and metastasis. Our data also showed that downregulation of TLR4 expression using an RNA silencing approach in A549 tumor cells significantly suppressed cell proliferation, cell migration and cell invasion, and induced tumor apoptosis in vitro, and suppressed tumor growth in vivo. In addition, we also found that downregulation of TLR4 expression significantly decreased cell TNF-α and IL-6 levels. Furthermore, we found that knockdown of TLR4 was able to significantly suppress constitutive phosphorylation of Akt and PI3K, which may contribute to the inhibition of tumor growth. These data suggest that TLR4 plays an important role in tumorigenic properties of human NSCLC, and that RNA interference-directed targeting of TLR4 could be used as a potential anticancer therapeutic target in NSCLC.

[Effects of acupotomy, electroacupuncture or round-sharp acupuncture needle interventions on expression of Bcl-2, Bax, Caspase-3 proteins of rectus femoris in rabbits with knee ostarthritis].

OBJECTIVE: To observe the effect of acupotomy, electroacupuncture (EA) or round-sharp acupuncture needle intervention on the expression of Bcl-2,Bax and Caspase-3 proteins in the rectus femoris in rabbits with knee ostarthritis (KOA), so as to explore their mechanisms underlying improvement of braking-induced joint damage from the cellular apoptosis. METHODS: Forty-five New Zealand rabbits were equally and randomized into control group, model group, acupotomy (AP) group, EA group and round-sharp acupuncture needle (RSAN) group (n = 9 in each group). The knee-joint injury model was established by fixing the left knee joint in extention position with plaster bandage. EA (2 Hz/100 Hz, 3 mA, 20 min each time) was applied to the left "Yanglingquan" (GB 34)- "Yinlingquan" (SP 9) and left "Neixiyan" (EX-LE 4)- "Waixiyan"(ST 35) for rabbits in the EA group. The EA treatment was given once daily, 3 times a week, 3 weeks in total. For rabbits of the AP group, a needle-knife was held to insert into the front edge of the midpoint, the starting point and the stopping point of the left medial collateral ligamen, lateral collateral ligament and the patellar ligament of the knee to make a loosening manipulation for 5 times in a session of treatment, once a week, 3 times altogether. For rabbits of the RSAN group, a round-sharp needle was performed in the same way to the needle-knife including the stimulation point, the manipulation method and treatment sessions. At the end of the experiment, the left rectus femoris was taken out for detecting the expression of Bcl-2, Bax and Caspase-3 proteins with Western blot. RESULTS: In comparison with the control group, the passive range of motion (PROM) level was significantly decreased 4, 8 and 12 weeks after modeling (P < 0.01), and the expression levels of Bax and Caspase-3 proteins in the rectus femoris were considerably upregulated in the model group (P < 0.05), while the ratio of Bcl-2/Bax was notably down-regulated (P < 0.05) in the model group. Compared with the model group, the PROM level at week 12 after modeling in the AP, EA and RSAN groups were significantly increased (P < 0.01); while Bax and Caspase-3 expression levels in both AP and RSAN groups were considerably downregulated (P < 0.05). No significant differences were found among the five groups in Bcl-2 expression levels (P > 0.05), and between the EA and model groups in Bax and Caspase-3 expression levels and the ratio of Bcl-2/Bax (P > 0.05). CONCLUSION: AP, RSAN and EA interventions are effective in improving the knee-joint motion range in KOA rabbits, and this effect of both AP and RSAN is closely associated with their actions in lowering the expression of Bax and Caspase-3 proteins of the rectus femoris and in raising ratio of Bcl-2/Bax protein (reducing muscular cellular apoptosis). The mechanism of EA intervention in improving PROM may be different.