SMAD4 regulates the progression of cholangiocarcinoma by modulating the expression of STING1.

SMAD4 is a tumour suppressor and an important regulator of tumour immune scape which is downregulated in cholangiocarcinoma (CCA). STING1 is a vital sensing factor of abnormal DNA; however, the correlation between SMAD4 and STING1 and the role of the SMAD4-STING1 interaction in the progression of CCA have not yet been evaluated. Public database was analysed to reveal the expression of SMAD4 and STING1. A cohort comprising 50 iCCA, 113 pCCA and 119 dCCA patients was assembled for the study. Immunohistochemistry was employed to evaluate the expression levels of STING1 and SMAD4. In vitro transwell and CCK8 assays, along with luciferase reporter assay, were conducted to analyse the potential regulatory mechanisms of SMAD4 on the expression of STING1. Expression of SMAD4 and STING1 were downregulated in CCA tumours and STING1 expression correlated with SMAD4 expression. The overexpression of SMAD4 was found to suppress the migration, invasion and proliferation capabilities of CCA cells; whereas, the knockdown of SMAD4 enhanced these abilities. Furthermore, it was observed that SMAD4 translocated into the nucleus following TGF-ß1 stimulation. Knockdown of SMAD4 resulted in the inhibition of STING1 transcriptional activity, whereas the overexpression of SMAD4 promoted the transcriptional activity of STING1. Clinically, low STING1 and SMAD4 expression indicated poor prognosis in CCA, and simultaneously low expression of STING1 and SMAD4 predicts poorer patient survival. SMAD4 regulates the expression of STING1 through its transcription regulating function. Dual low expression of STING1 and SMAD4 had more power in predicting patient survival. These results indicate that SMAD4-silenced CCA may downregulate its STING1 expression to adapt to the immune system.

Case report: Obstructive jaundice caused by biliary cystadenoma.

Biliary cystadenoma (also called mucinous cystic neoplasm with low-grade intraepithelial neoplasia) is a rare cystic tumor that arises from the biliary epithelium. The cause of biliary cystadenoma is still unclear. Jaundice is a rare presentation of intrahepatic biliary cystadenoma, which can lead to a diagnostic dilemma. Herein, we present a case of intrahepatic biliary cystadenoma that primarily exhibited as jaundice. A 56-year-old woman has suffered from yellow staining of her skin and sclera for more than 1 month. She had a poor appetite and mild epigastric pain. Laboratory examination showed elevated levels of total bilirubin and elevated carbohydrate antigen 19-9 (CA19-9). A contrast-enhanced computed tomography of the abdomen showed a 7.4 * 5.3-cm, oval, low-density lesion in the left liver parenchyma with a clear boundary and visible septa. The common bile duct was obviously dilated with wall thickening. On magnetic resonance imaging, the lesion in the liver showed a multilocular cystic, unenhanced long T2 signal. There was local thickening of the common bile duct wall with short T2-like filling defects and high signal intensity on diffusion-weighted imaging (DWI). The patient had no history of other malignant tumors and adjuvant therapy such as radiotherapy and chemotherapy. She was clinically suspected of having either biliary cystadenoma or a malignancy; hence, resection was performed. Macroscopically, the excised tissue specimen showed a polypoid mass in the common bile duct, which extended along the bile duct to the intrahepatic bile duct. There was a cystic and solid mass in the left liver with yellow turbid fluid, which was associated with the polypoid mass in the common bile duct. Histopathology suggests mucinous cystadenoma of the liver and hilar bile duct. The differential diagnosis of biliary cystadenoma and treatment selection have been discussed.

Advances in molecular and cell therapy for immunotherapy of cholangiocarcinoma.

Cholangiocarcinoma (CCA) is a highly malignant tumor of the hepatobiliary system that has failed to respond to many traditional therapies to a certain extent, including surgery, chemotherapy and radiotherapy. In recent years, the new therapeutic schemes based on immunology have fundamentally changed the systemic treatment of various malignant tumors to a certain extent. In view of the immunogenicity of CCA, during the occurrence and development of CCA, some immunosuppressive substances are released from cells and immunosuppressive microenvironment is formed to promote the escape immune response of its own cells, thus enhancing the malignancy of the tumor and reducing the sensitivity of the tumor to drugs. Some immunotherapy regimens for cholangiocarcinoma have produced good clinical effects. Immunotherapy has more precise characteristics and less adverse reactions compared with traditional treatment approaches. However, due to the unique immune characteristics of CCA, some patients with CCA may not benefit in the long term or not benefit at all after current immunotherapy. At present, the immunotherapy of CCA that have been clinically studied mainly include molecular therapy and cell therapy. In this article, we generalized and summarized the current status of immunotherapy strategies including molecular therapy and cell therapy in CCA in clinical studies, and we outlined our understanding of how to enhance the clinical application of these immunotherapy strategies.

Research progress of bile biomarkers and their immunoregulatory role in biliary tract cancers.

Biliary tract cancers (BTCs), including cholangiocarcinoma and gallbladder carcinoma, originate from the biliary epithelium and have a poor prognosis. Surgery is the only choice for cure in the early stage of disease. However, most patients are diagnosed in the advanced stage and lose the chance for surgery. Early diagnosis could significantly improve the prognosis of patients. Bile has complex components and is in direct contact with biliary tract tumors. Bile components are closely related to the occurrence and development of biliary tract tumors and may be applied as biomarkers for BTCs. Meanwhile, arising evidence has confirmed the immunoregulatory role of bile components. In this review, we aim to summarize and discuss the relationship between bile components and biliary tract cancers and their ability as biomarkers for BTCs, highlighting the role of bile components in regulating immune response, and their promising application prospects.

Analysis of Key Genes for Slow Transit Constipation Based on RNA Sequencing.

Purpose: This study aims to identify key genes in slow transit constipation (STC). We also sought to explore the potential link between STC and colorectal cancer. Patients and Methods: mRNA expression profiles were obtained by RNA sequencing, and differentially expressed genes were identified. Functional enrichment analysis and a protein-protein interaction (PPI) network was explored, and differentially expressed genes common to STC and colorectal cancer were examined. Analysis of the effect of constipation and colorectal cancer common genes on the overall survival of colorectal cancer patients based on GEPIA database. Results: Functional enrichment showed that significantly different genes are related to lymphocyte chemotaxis, positive regulation of inflammatory response, cellular response to tumor necrosis factor, extracellular region, extracellular space and chemokine activity. The hub gene for STC was found in the PPI network. In addition, AQP8 and CFD were common differential genes for STC and colorectal cancer. AQP8 affects overall survival in patients with colorectal cancer. Conclusion: Our findings will contribute to understanding the pathology of STC at the molecular level, with the first discovery that AQP8 may be a hub gene in the transition from STC to colorectal cancer.

A Severe Cervical Spondylotic Myelopathy Patient Treated with Integrative Korean Medicine Including Acupuncture, Bee Venom Pharmacopuncture and Herbal Medicine: a case report.

Cervical spondylotic myelopathy (CSM) is common in elderly people and severe CSM patients are recommended to receive surgery. However, in some cases, surgery may fail to improve the patients' symptoms. An 80-year-old man diagnosed with CSM complained of right hemiplegia and right arm and leg pain with the presence of a Foley catheter, despite treatment with laminectomy and laminoplasty. Acupuncture, bee venom pharmacopuncture, and herbal medicine were administered for 129 days. As a result, manual muscle testing (MMT) and the Modified Barthel Index (MBI) improved, the pain in his right arm and leg decreased, and he was able to urinate by himself. This case report implies that integrative Korean medicine (IKM) can be an option for patients suffering from muscular weakness resulting from myelopathy.

Human gut-microbiome-derived propionate coordinates proteasomal degradation via HECTD2 upregulation to target EHMT2 in colorectal cancer.

The human microbiome plays an essential role in the human immune system, food digestion, and protection from harmful bacteria by colonizing the human intestine. Recently, although the human microbiome affects colorectal cancer (CRC) treatment, the mode of action between the microbiome and CRC remains unclear. This study showed that propionate suppressed CRC growth by promoting the proteasomal degradation of euchromatic histone-lysine N-methyltransferase 2 (EHMT2) through HECT domain E3 ubiquitin protein ligase 2 (HECTD2) upregulation. In addition, EHMT2 downregulation reduced the H3K9me2 level on the promoter region of tumor necrosis factor α-induced protein 1 (TNFAIP1) as a novel direct target of EHMT2. Subsequently, TNFAIP1 upregulation induced the apoptosis of CRC cells. Furthermore, using Bacteroides thetaiotaomicron culture medium, we confirmed EHMT2 downregulation via upregulation of HECTD2 and TNFAIP1 upregulation. Finally, we observed the synergistic effect of propionate and an EHMT2 inhibitor (BIX01294) in 3D spheroid culture models. Thus, we suggest the anticancer effects of propionate and EHMT2 as therapeutic targets for colon cancer treatment and may provide the possibility for the synergistic effects of an EHMT2 inhibitor and microbiome in CRC treatment.

Prognostic impact of serum soluble PD-1 and ADV score for living donor liver transplantation in patients with previously untreated hepatocellular carcinoma.

PURPOSE: The programmed death protein 1 (PD-1) pathway is the critical mechanism in development of hepatocellular carcinoma (HCC). The present study analyzed the prognostic impact of pretransplant serum soluble PD-1 (sPD-1) concentration and α-FP-des-γ-carboxyprothrombin-tumor volume (ADV) score in patients with previously untreated HCC undergone liver transplantation (LT). METHODS: This retrospective single-center study enrolled 100 patients with HCC who underwent living donor LT from 2010 to 2016. Concentrations of sPD-1 were measured in stored serum samples. RESULTS: Receiver operating characteristic curve analysis of 2-year tumor recurrence resulted in an sPD-1 cutoff of 177.1 µg/mL, which was associated with higher rates of tumor recurrence (P = 0.022), but not with overall patient survival (P = 0.460). The derived cutoff for pretransplant ADV score was 5.4log, which was associated with higher tumor recurrence rate (P < 0.001) and lower overall patient survival rate (P < 0.001). Both sPD-1 of >177.1 µg/mL (hazard ratio [HR], 2.26; P = 0.020) and pretransplant ADV score of >5.4log (HR, 3.56; P < 0.001) were independent risk factors for posttransplant HCC recurrence. The combination of these 2 factors enabled the stratification of patients into 3 groups, with groups having 0, 1, and 2 risk factors differing significantly in the prognosis of tumor recurrence (P < 0.001) and overall patient survival (P = 0.006). CONCLUSION: Both sPD-1 concentration and ADV score have prognostic impacts in patients who underwent LT for untreated HCCs. These factors, both individually and combined, can help in predicting posttransplant prognosis.

[Investigation and analysis of perioperative nutritional status in patients with malignant oral and maxillofacial neoplasms complicated with diabetes mellitus].

PURPOSE: To analyze the nutritional status of patients with malignant oral and maxillofacial neoplasms complicated with diabetes mellitus during perioperative period. METHODS: Sixty-four patients with malignant oral and maxillofacial neoplasms complicated with diabetes mellitus were collected. Fasting venous blood of patients was extracted on the morning of the day before surgery and then at the 1st, 3rd and 7th day after surgery, respectively. The hemoglobin (Hb), total protein (TP), pre-albumin (PA), lymphocyte count (L), creatinine (Scr), uric acid (UA), urea nitrogen (BUN), glomerular filtration rate (EGFR_EPI_C), white blood cells (WBC) of the patients were detected. The intake of enteral nutrition on the 7th day after operation was investigated. The correlation between nutritional status and the length of stay was analyzed. SPSS 23.0 software package was used for statistical analysis. RESULTS: Compared with the indicators before surgery, the values of TP, ALB, PA, Hb were decreased significantly after surgery in all groups(P<0.05). The level of TP on postoperative day 7 was significantly higher than that on postoperative day 1 and postoperative day 3(P<0.05). The PA level on the third day after surgery was significantly lower than that on the first day after surgery(P<0.05). The Hb level on the 7th and 3rd day after surgery was significantly lower than that on the 1st day after surgery (P<0.05). The level of L decreased significantly in each group after surgery compared with the level before surgery(P<0.05), and gradually increased in each group after surgery, with significant difference among each two groups (P<0.05). Compared with preoperative value, blood Scr was significantly decreased in all groups after surgery (P<0.05), the UA level after surgery was significantly lower than the preoperative level in all groups(P<0.05), and at the 3rd day after surgery was significantly lower than at the 1st day after surgery(P<0.05). After surgery, the eGFR_EPI_c level was significantly higher than the level before surgery in all groups (P<0.05), and at the 7th day after surgery was significantly higher than at the 1st day after surgery (P<0.05). The level of WBC in all patients after surgery was significantly higher than that before surgery(P<0.05), and the level at the 3rd and 7th day after surgery was significantly higher than that at the 1st day after surgery(P<0.05). At the 7th day after surgery, the energy and protein intakes of the patients were significantly lower than the recommendations. There was positive correlation between preoperative BMI and TP, ALB levels at the 1st postoperative day and the TP level at 3rd postoperative day(P<0.05). There was no direct correlation between preoperative BMI and the length of postoperative hospital stay (P>0.05). The length of postoperative hospital stay was negatively correlated with the age and negatively with TP and ALB levels at the 1st postoperative day(P<0.05). CONCLUSIONS: Early postoperative nutritional status of patients with malignant oral and maxillofacial neoplasms complicated with diabetes mellitus decreased significantly. The energy and protein intakes of the patients are significantly lower than the recommendations. The length of postoperative hospital stay is negatively correlated with early postoperative nutritional status and age.

A novel recombinase polymerase amplification assay for rapid detection of epidemic fowl adenovirus.

Fowl adenovirus (FAdV) has posed a grave threat to the health of poultry, and the sudden outbreak highlights the importance of the new rapid diagnostic method for the control and prevention of transmission. Hence, in the present study, a novel recombinase polymerase amplification (RPA) assay, which was suitable for all 12 serotypes (FAdV-1 to 8a and 8b to 11) had been successfully launched to detect FAdV. Also, the entire amplification process could be completed in the isothermal condition when temperature ranged from 26 to 42°C within no more than 14 min, which was remarkably superior to endpoint polymerase chain reaction (98 min) with the same detecting sensitivity (as low as 0.1 fg viral DNA), avoiding sophisticated thermal cyclers with simple operation. Additionally, the same primers did not produce positive reactions with other viruses tested, demonstrating that the specificity of the RPA assay was acceptable. Moreover, this developed method could be efficiently used in the diagnosis of FAdV references and epidemic strains from different avian origins, thus making it a rapid, reliable, and point-of-care FAdV diagnostics tool, as well as an alternative to endpoint PCR.

Cereblon Deletion Ameliorates Lipopolysaccharide-induced Proinflammatory Cytokines through 5'-Adenosine Monophosphate-Activated Protein Kinase/Heme Oxygenase-1 Activation in ARPE-19 Cells.

Cereblon (CRBN), a negative modulator of AMP-activated protein kinase (AMPK), is highly expressed in the retina. We confirmed the expression of CRBN in ARPE-19 human retinal cells by Western blotting. We also demonstrated that CRBN knock-down (KD) could effectively downregulate IL-6 and MCP-1 protein and gene expression in LPS-stimulated ARPE-19 cells. Additionally, CRBN KD increased the phosphorylation of AMPK/acetyl-coenzyme A carboxylase (ACC) and the expression of heme oxygenase-1 (HO-1) in ARPE-19 cells. Furthermore, CRBN KD significantly reduced LPS-induced nuclear translocation of NF-κB p65 and activation of NF-κB promoter activity. However, these processes could be inactivated by compound C (inhibitor of AMPK) and zinc protoporphyrin-1 (ZnPP-1; inhibitor of HO-1). In conclusion, compound C and ZnPP-1 can rescue LPS-induced levels of proinflammatory cytokines (IL-6 and MCP-1) in CRBN KD ARPE-19 cells. Our data demonstrate that CRBN deficiency negatively regulates proinflammatory cytokines via the activation of AMPK/HO-1 in the retina.

[Correlation between nutritional status before radiotherapy and dietary intake in patients with oral cancers].

PURPOSE: To explore the correlation between dietary nutrient intake and PG-SGA score in patients with oral cancers before radiotherapy. METHODS: Sixty-five patients with oral cancers treated in Shanghai Ninth People's Hospital were selected. The 72-hour dietary survey method was used to understand the food intake of the patients. PG-SGA was used to make quantitative scoring of the nutritional status of the patients. SPSS 23.0 software package was used for data analysis. RESULTS: The number of severe malnutrition cases in male patients was significantly higher than that in female patients (P＜0.05). Energy, fat and fat energy ratio, carbohydrate and carbohydrate energy ratio from diet in good nutrition/mild malnutrition group, moderate malnutrition, severe malnutrition patients were over reference intake of dietary nutrients for Chinese residents; protein intake and protein energy ratio were greater than reference intake of nutrients for Chinese people. There was no significant difference among the three groups. The selenium intake of the well-nourished/mildly malnourished, moderately malnourished and severely malnourished groups was 67.15, 81.04 and 81.59 µg; vitamin E was 27.81, 30.88, 26.40 mg α-TE; vitamin C was 150.19, 159.81, 183.71 mg; retinol was 904.65, 1401.51, and 1373.81 µg RAE, respectively. Niacin was 12.97, 18.76 and 14.27 mg NE, respectively, reaching or exceeding the reference intake. There was no significant difference among the three groups. In male patients, dietary energy and niacin intake were negatively correlated with PG-SGA score (P<0.05). CONCLUSIONS: Patients with oral cancers have a high incidence of malnutrition before radiotherapy, and the average intake of dietary energy, protein, fat, carbohydrates and micronutrients such as selenium, vitamin E, vitamin C, retinol and niacin reached or exceeded the reference intake. Energy and niacin intake were negatively correlated with PG-SGA score in male patients.

[Clinical application of magnifying colonoscopy in diagnosis and treatment of colorectal benign neoplastic lesions].

OBJECTIVE: To evaluate the clinical value of magnifying endoscopy in diagnosis and treatment of colorectal benign neoplastic lesions. METHODS: Seventy-eight colorectal lesions in 61 patients were examined with magnifying colonoscopy after indigo carmine dyeingìand pit pattern diagnosis was made for every lesion according to Kudos classification to differentiate neoplastic lesions from non-neoplastic lesions. The lesions were resected by endoscopic polypectomy and mucosectomy or surgical treatment. The diagnoses made by magnifying colonoscopy were compared with pathologic results. RESULTS: The diagnostic sensitivity of magnifying endoscopy for neoplastic lesions was 98.4% and specificity was 85.7%. The overall accuracy for adenoma and early colorectal cancer was 96.2%. 89.7% of adenomatous lesions were treated by endoscopic resection. CONCLUSION: The magnifying colonoscopy can provide instantaneous and accurate diagnosis of neoplastic lesions in colon and rectum,as well as minimally invasive treatment.