Adenocarcinoma developing from gastric heterotopic pancreas: a case report and short review.

Heterotopic pancreas is a relatively rare condition that may be associated to clinical complaints or signs. Here, we report a case of gastric heterotopic pancreas assictaed to ductal adenocarcinoma. Obstructive jaundice was the initial symptom prompting medical intervention. A 73-year-old male patient presented with yellow staining of the skin and sclera, and dull epigastric pain. Contrast-enhanced computed tomography showed stenosis of the extrahepatic distal bile duct and mass lesions of the antrum. The patient underwent tumor resection, distal gastrectomy (Billroth II), and common bile duct exploration. Postoperative pathological examination revealed an adenocarcinoma located in the wall of the gastric antrum. Immunohistochemical results suggested that the tumor originated from the pancreas. Heterologous pancreatic tissue and a dilated pancreatic duct were found in the tumor. These findings suggest malignant transformation of the gastric heterotopic pancreas. Of note, jaundice as clinical complaint for adenocarcinoma associated to gastric heterotopic pancreas.

Role of connexin 32 in the directional differentiation of induced pluripotent stem cells into hepatocytes.

This study aimed to explore the role of connexin 32 (Cx32) in the directional differentiation of induced pluripotent stem cells (iPSCs) into hepatocytes. Urine-derived epithelial cells were collected from the fresh urine of a healthy donor and transducted with reprogramming plasmid mixture to generate iPSCs. The iPSCs were then directionally differentiated into hepatocytes. During the differentiation, the upregulated and downregulated groups were treated with vitamin K2 (VK2) and 2-aminoethoxyboronate diphenylester (2-APB) to increase and inhibit Cx32 expression, respectively. The control group was not treated with the regulatory factor. Expression of Cx32 and hepatocyte-specific markers, including AFP, hepatocyte nuclear factor 4α (HNF-4α), albumin (ALB) and cytokeratin 18 (CK18) were detected. It indicated that Cx32 expression was not observed in iPSCs, but gradually increased during the process of hepatic differentiation from iPSCs. Upregulation of Cx32 expression by VK2 treatment promoted hepatocyte maturation and enhanced the expression of the aforementioned hepatic specific markers, whereas downregulation of Cx32 expression by 2-APB treatment had the opposite effects. In conclusion, urine-derived iPSCs could be directionally differentiated into hepatocytes. Up-regulation of Cx32 improves the efficiency and maturity of differentiation of iPSCs into hepatocytes, and Cx32 may be a promoting factor during the process of hepatic differentiation from iPSCs.

Relationship between postoperative biliary stricture and clinical characteristics of patients with benign and malignant biliary diseases.

Introduction: Postoperative biliary stricture (POBS) is one of the common complications of biliary surgery. Previous literature on risk factors of POBS was scarce, and the classification of POBS in benign and malignant biliary diseases was incomplete. Aim: To analyze clinicopathological factors of POBS in usual biliary diseases, and to facilitate preoperative diagnosis of biliary stricture. Material and methods: A retrospective analysis was made on the clinical data of 2228 patients who underwent biliary surgery in our hospital from July 2010 to June 2022. With the inclusion and exclusion criteria, the clinicopathological factors for POBS were classified, and data analysis was conducted. Results: Benign diseases with age ≥ 60 years (p = 0.034), diabetes (p = 0.001), common bile duct diameter < 0.8 cm (p = 0.034), Mirizzi syndrome (p = 0.001), seniority of surgeons < 25 years (p = 0.001), and operation time for the first 6 years (p = 0.015) are more likely to evolve into POBS; malignant diseases with conjugated bilirubin ≥ 6.8 µmol/l (p = 0.042), alkaline phosphatase ≥ 125 U/l (p = 0.042), γ-glutamyl transferase ≥ 50 U/l (p = 0.047), diabetes (p = 0.038), and seniority of surgeons < 25 years (p = 0.008) are prone to POBS. Different surgical approaches affect the incidence of POBS (χ2 = 9.717, p = 0.034). The choice of surgical site is important for the incidence of POBS in malignant diseases (χ2 = 7.935, p = 0.041). Conclusions: Surgeons need to identify risk factors, conduct patient visits and assessments preoperatively, standardize the operation in order to avoid structural damage, and reduce the occurrence of POBS.

Cancer-associated fibroblasts suppress ferroptosis and induce gemcitabine resistance in pancreatic cancer cells by secreting exosome-derived ACSL4-targeting miRNAs.

BACKGROUND: Pancreatic cancer continues to be one of the world's most lethal cancers. Chemotherapy resistance in patients with advanced pancreatic cancer often accompany with dismal prognosis, highlighting the need to investigate mechanisms of drug resistance and develop therapies to overcome chemoresistance. METHODS: This research was filed with the Chinese Clinical Trial Registry (ChiCTR2200061320). In order to isolate primary normal fibroblasts (NFs) and cancer-associated fibroblasts (CAFs) samples of pancreatic ductal adenocarcinoma (PDAC) and paracancerous pancreatic tissue from individuals diagnosed with PDAC were obtained. The exosomes were obtained using ultracentrifugation, and their characteristics were determined by Western blotting, nanoparticle tracking analysis, and transmission electron microscopy. CAF-derived miRNAs were analyzed by RT-qPCR and high-throughput sequencing. Gemcitabine (GEM) was employed to promote ferroptosis, and ferroptosis levels were determined by monitoring lipid reactive oxygen species (ROS), cell survival, and intracellular Fe2+ concentrations. To assess in vivo tumor response to GEM therapy, a xenograft tumor mouse model was utilized. RESULTS: Exosomes derived from CAFs in PDAC did not exhibit innate GEM resistance. CAFs promoted chemoresistance in PDAC cells following GEM treatment by secreting exosomes, and maintaining signaling communication with cancer cells. Mechanistically, miR-3173-5p derived from CAF exosomes sponged ACSL4 and inhibited ferroptosis after uptake by cancer cells. CONCLUSION: This work demonstrates a novel mode of acquired chemoresistance in PDAC and identifies the miR-3173-5p/ACSL4 pathway as a promising treatment target for GEM-resistant pancreatic cancer.

Acute diffuse peritonitis secondary to a seminal vesicle abscess: A case report.

BACKGROUND: Seminal vesicle abscess (SVA) is the manifestation of a relatively rare urinary system infection. In response to urinary system inflammation, an abscess forms in special locations. However, acute diffuse peritonitis (ADP) induced by SVA is unusual. CASE SUMMARY: We report a case of a left SVA in a male patient complicated with pelvic abscess, ADP, multiple organ dysfunction syndrome, infectious shock, bacteremia, and acute appendiceal extraserous suppurative inflammation as a result of a long-term indwelling urinary catheter. The patient received a course of morinidazole + cefminol antibiotics but showed no obvious relief, so the perineal SVA underwent puncture drainage and abdominal abscess drainage + appendectomy was performed. The operations were successful. After the operation, anti-infection, anti-shock, and nutritional support treatments were continued and various laboratory indicators were regularly reviewed. The patient was discharged from the hospital after recovery. This disease is a challenge for the clinician because of the unusual spreading path of the abscess. Moreover, appropriate intervention and adequate drainage of abdominal and pelvic lesions are necessary, especially when the primary focus cannot be determined. CONCLUSION: The etiology of ADP varies, but acute peritonitis secondary to SVA is very rare. In this patient, the left SVA not only affected the adjacent prostate and bladder but also spread retrogradely through the vas deferens, forming a pelvic abscess in the loose tissues of the extraperitoneal fascia layer. Inflammation involving the peritoneal layer led to ascites and pus accumulation in the abdominal cavity, and appendix involvement led to extraserous suppurative inflammation. In clinical practice, surgeons need to consider the results of various laboratory tests and imaging examinations to make comprehensive judgments involving the diagnosis and treatment plan.

Improving the Therapeutic Efficacy of Sorafenib for Hepatocellular Carcinoma by Repurposing Disulfiram.

Background: Sorafenib, a kinase inhibitor, is a standard treatment for advanced hepatocellular carcinoma (HCC) but provides only a limited survival benefit. Disulfiram (DSF), a drug for treating alcoholism and a chelator of copper (Cu), forms a complex with Cu (DSF/Cu). DSF/Cu is a potent inducer of autophagic apoptosis of cancer stem cells, which can demonstrate drug resistance. Thus, we hypothesized that DSF/Cu could increase the sensitivity of HCC cells to sorafenib by targeting hepatic cancer stem cells. Methods: The synergistic effect of DSF/Cu and sorafenib on human HCC cell lines was assessed by cell viability MTT assay. Changes in stemness gene expression in HCC cells were investigated by assessing the presence of hepatic cancer stem cells (HCSCs) (defined as ALDH+ cells) using flow cytometry, sphere formation ability as an index of in vitro tumorigenicity, and expression of stemness gene-encoded proteins by western blot. Autophagic apoptosis and the ERK signaling pathway were also assessed by western blot. Most importantly, the in vivo anti-tumor efficacy of DSF/Cu and sorafenib was tested using orthotopic HCC xenografts in mice. Results: Compared with sorafenib alone, DSF/Cu + sorafenib synergistically inhibited proliferation of all HCC cell lines, decreased the stemness of HCC cells, and increased the autophagy and apoptosis of HCC cells. The mechanism by which DSF/Cu mediated these phenomena with sorafenib was sustained activation of the ERK pathway. The combination of DSF/Cu (formed with endogenous Cu2+) and sorafenib was significantly more effective than sorafenib alone in inhibiting the growth of orthotopic HCC xenografts in mice. This in vivo anti-tumor efficacy was associated with decreased stemness in treated HCC tumors. Conclusions: DSF/Cu and sorafenib can synergistically and effectively treat HCC by targeting HCSCs in vitro and in vivo. Our data provide a foundation for clinical translation.

The role of non-coding RNAs in ferroptosis regulation.

Ferroptosis is a newly recognized form of cell death that is distinct from apoptosis, necrosis, autophagy in morphology, biochemistry, and heredity. The basic process of ferroptosis involves disordered permeability of plasma membrane, which is caused by abnormal accumulation of lipids and reactive oxygen species (ROS). The regulatory mechanism of ferroptosis is important due to its involvement in tumor progression, neurotoxicity, neurodegenerative diseases, acute renal failure, and ischemia-reperfusion injury. Recent studies have shown that in ferroptosis metabolism, non-coding RNAs (ncRNAs) can interfere with multiple signaling pathways at both the pre-transcriptional and post-transcriptional levels. Despite great progress, current research on the mechanism of ncRNAs and ferroptosis remains insufficient. This review provides an overview of the main mechanisms and targets of ferroptosis and focuses on the mechanisms of non-coding RNA regulation. Analyzing the deficiencies in current research may provide ideas for future studies to investigate.

Asymptomatic gastric adenomyoma and heterotopic pancreas in a patient with pancreatic cancer: A case report and review of the literature.

BACKGROUND: Gastric adenomyoma (GA) is a rare submucosal benign neoplasm that occurs mostly in the gastric antrum and is often misdiagnosed. No standard treatment has been established for this disease in cases of malignancy. CASE SUMMARY: A 75-year-old woman with a 10-year history of hypertension was admitted to the Emergency Department of our hospital complaining of paroxysmal exacerbation of acute abdominal pain for 1 d with no apparent cause. Enhanced computed tomography and magnetic resonance imaging indicated a mass in the caudal pancreas, cholecystitis, and cholecystic polypus. Gastrointestinal endoscopy showed a mass arising from the gastric antrum. Due to the imaging findings, pancreatic cancer (PC), gastric lesion, cholecystitis, and cholecystic polypus were our primary consideration. Radical pancreatectomy, splenectomy, and cholecystectomy were performed successfully, and the gastric tumor was locally resected. Postoperative paraffin specimens confirmed the diagnosis of caudal PC, GA, and heterotopic pancreas (HP). Unfortunately, the patient died 13 mo later due to PC metastases to the liver, lung, and adrenal glands. CONCLUSION: GA is a rare benign disease, especially when occurring with HP. It may stem from the same origin as HP. This is the first case report to date of a patient suffering from the simultaneous occurrence of GA, HP, and PC. GA is a lesion that can mimic other benign or malignant gastrointestinal diseases; thus, a definitive diagnosis depends on postoperative pathological biopsy. Although GA and HP are both benign lesions, they should be resected because there is a chance of malignancy. Additional research should be conducted to better understand these submucosal lesions.

Rupture and hemorrhage of a seminoma mixed with yolk sac tumors in 46XY partial gonadal dysgenesis: a case report and literature review.

BACKGROUND: 46XY partial gonadal dysgenesis (PGD) is a rare subtype of disorder of sex development (DSD). 46YY PGD is a congenital disease with atypical chromosomal, gonadal, or anatomical sex development. The patient in this case report had male and female genitalia simultaneously. We created a flowchart of the differential diagnosis for clinicians. CASE PRESENTATION: A 41-year-old male was admitted to the hospital complaining of lower quadrant abdominal pain for 1 day. Physical examination revealed that his penis size was normal, but a urethral orifice was located in the perineum area between the scrotum and anus. One small testicle was in the left scrotum, but no testicle was present on the right. The patient's abdomen was bulging, and he had lower abdominal pain. According to the emergency CT scan, a lesion (74*65 mm) was found in the right pelvis between the bladder and rectum. The lesion showed an unclear boundary and hematocele appearance. The lesion was removed by emergency surgery, and the pathology report indicated a mixed germ cell tumor with a seminoma and yolk sac tumors. CONCLUSION: This article is a case report of germ cell tumors in 46XY PGD patients. The literature review summarizes the clinical diagnosis, and a flowchart is provided for physicians in future practice. The importance of this report is that it will help acquaint physicians with this rare disease and make the right initial clinical decision quickly through the use of this flowchart. However, the variants of special subtypes of 46XY DSD are myriad, and all the diagnoses could not be covered in one flowchart.

TRPM2 promotes pancreatic cancer by PKC/MAPK pathway.

The mechanism of pancreatic cancer (PA) is not fully understanded. In our last report, TRPM2 plays a promising role in pancreatic cancer. However, the mechanism of TRPM2 is still unknown in this dismal disease. This study was designed to investigate the role and mechanism of TRPM2 in pancreatic cancer. TRPM2 overexpressed and siRNA plasmid were created and transfected with pancreatic cancer cell line (BxPC-3) to construct the cell model. We employed CCK-8, Transwell, scratch wound, and nude mice tumor-bearing model to investigate the role of TRPM2 in pancreatic cancer. Besides, we collected the clinical data, tumor tissue sample (TT) and para-tumor sample (TP) from the pancreatic cancer patients treated in our hospital. We analyzed the mechanism of TRPM2 in pancreatic cancer by transcriptome analysis, western blot, and PCR. We blocked the downstream PKC/MEK pathway of TRPM2 to investigate the mechanism of TRPM2 in pancreatic cancer by CCK8, scratch wound healing, and transwell assays. Overexpressed TRPM2 could promote pancreatic cancer in proliferation, migration, and invasion ability in no matter the cell model or nude mice tumor-bearing model. TRPM2 level is highly negative correlated to the overall survival and progression-free survival time in PA patients, however, it is significantly increased in PA tissue as the tumor stage increases. The transcriptome analysis, GSEA analysis, western-blot, and PCR results indicate TRPM2 is highly correlated with PKC/MAPK pathways. The experiments of PKC/MEK inhibitors added to TRPM2 overexpressed BxPC-3 cell showed that significant inhibition of PA cells happened in CCK8, transwell, and wound-healing assay. TRPM2 may directly activate PKCα by calcium or indirectly activate PKCε and PKCδ by increased DAG in PA, which promote PA by downstream MAPK/MEK pathway activation.

Human Mesenchymal Stem Cell-Derived Exosomal microRNA-143 Promotes Apoptosis and Suppresses Cell Growth in Pancreatic Cancer via Target Gene Regulation.

BACKGROUND: This study aimed to explore the regulatory mechanism of hsa-miR-143-3p and lncRNA RP11-363N22.3-functioning upstream of KRAS-in exosomes derived from human mesenchymal stem cells (hMSCs) in pancreatic cancer. METHODS: Western blotting and quantitative PCR were used to determine gene expression. In vitro, cell proliferation, apoptosis, and cell cycle and invasion were evaluated using CCK-8 assay, flow cytometry, and transwell assays, respectively. In vivo, the effect of hsa-miR143-3p was investigated using a tumorigenesis test in nude mice. The association between hsa-miR-143-3p and lncRNA RP11-363N22.3 was investigated using the dual-luciferase assay. RESULTS: hsa-miR-143-3p expression significantly increased in hMSC exosomes than in those in human pancreatic cancer cell line (CFPAC-1) exosomes. In vitro, compared to the MOCK (CFPAC-1 only) group, cell proliferation and invasion were inhibited and apoptosis was induced in the inhibitor NC (CFPAC-1 + MSC-hsa-miR-3p inhibitor NC) group, while these changes were reversed in the inhibitor (CFPAC-1 + MSC-hsa-miR-3p inhibitor) group. The expression of lncRNA RP11-363N22.3 and genes related to miR-143 significantly decreased in the inhibitor NC group compared to the MOCK group, and increased in the inhibitor group compared to inhibitor NC group. A targeted combinatorial effect was observed between lncRNA RP11-363N22.3 and hsa-miR-143-3p. In vivo, the tumor volume of the mimics (CFPAC-1 + MSC-hsa-miR-143-3p mimics) group was smaller than that of the mimics NC (CFPAC-1 + MSC-hsa-miR-143-3p mimics NC) and MOCK groups. H&E staining showed that there were no obvious pathological changes in MOCK and mimic NC groups, while cell necrosis was seen in some regions in mimic groups. CONCLUSION: hsa-miR-143-3p may promote apoptosis and suppress cell growth and invasion in pancreatic cancer.

A FAK Inhibitor Boosts Anti-PD1 Immunotherapy in a Hepatocellular Carcinoma Mouse Model.

Anti-PD-1/PD-L1 immunotherapy has limited efficacy in hepatocellular carcinoma (HCC) and does not benefit all patients. A FAK inhibitor (VS-4718) has been reported to improve the microenvironment in some tumors. This study aimed to investigate the effect of the combination of the FAK inhibitor VS4718 and anti-PD1 for the treatment of HCC in a mouse model and its possible mechanism of action. The expression of FAK and infiltrated immune cells in human HCC from the data of TCGA were analyzed. A primary murine HCC model was established via protooncogene (c-Met/ß-catenin) transfection. The pathological characteristics of tumors were examined after the mice were treated with VS4718 and/or anti-PD1 therapy. This study revealed that FAK is highly expressed in human HCC and is associated with poor prognosis of OS (overall survival) and PFS (progress free survival) in HCC patients. Immune cell infiltration (CD8+ T, Tregs, M0, M2, CAFs and MDSCs) was correlated with FAK expression. In the experimental HCC model, the combination of a FAK inhibitor VS4718 and an anti-PD1 antibody had a better effect than monotherapy against HCC. VS4718 reduced the number of Tregs and macrophages but increased the number of CD8+ T cells in HCC mice. Notably, FAK inhibitor promoted the expression of PD-L1 in HCC. This study suggested that combination of the FAK inhibitor VS4718 and anti-PD1 could be a potential therapy for HCC by improving the immune environment, reducing liver fibrosis and simultaneously preventing PD1 from binding to the increased PD-L1 induced by FAK inhibitor VS4718.

Synchronous hepatocellular carcinoma and gallbladder adenocarcinoma with neuroendocrine differentiation: a case report and literature review.

BACKGROUND: Double primary cancers have a low incidence rate, and synchronous hepatocellular carcinoma and gallbladder adenocarcinoma are rarely reported. Here, we report such a case- the 12th case of synchronous double primary cancers featuring HCC and GC, but the first case of neuroendocrine differentiation in the gallbladder. CASE PRESENTATION: A 77-year-old female was admitted to the hospital complaining of weakness and inappetence for six months. Contrast-enhanced computed tomography (CT) of the abdomen indicated an 11 cm space-occupying lesion in the right lobe of the liver. Later, magnetic resonance imaging showed a high possibility of a massive hepatoma, and multiple gallstones were also seen. After transhepatic arterial chemoembolization, a repeat abdominal CT showed obvious local nodular thickening in the gallbladder wall. Finally, resection of the right lobe of the liver and cholecystectomy were performed. During an approximately 2-year follow-up, the patient recovered uneventfully without recurrence or metastasis. CONCLUSION: The disease in this case is rare and lacked typical radiological features. More precise and advanced diagnostic techniques are needed to obtain a clear diagnosis and refine treatment strategies. The management strategy should always be curative, even in the presence of multiple malignancies.

Anticancer effects of miR-124 delivered by BM-MSC derived exosomes on cell proliferation, epithelial mesenchymal transition, and chemotherapy sensitivity of pancreatic cancer cells.

OBJECTIVE: This study aims to explore the roles of miR-124 in pancreatic tumor and potential vehicles. RESULTS: The miR-124 expression levels decreased in pancreatic adenocarcinoma tissues and cancer cell lines AsPC-1, PANC1, BxPC-3 and SW1990. Furthermore, the elevated expression of miR-124 in AsPC-1 and PANC1 via miR-124 mimic transfection-induced apoptosis, metastasis and epithelial mesenchymal transition was suppressed, and the EZH2 overexpression partly reversed the protective effects of miR-124 against pancreatic tumors. In addition, the expression of miR-124 was detected in exosomes extracted from miR-124-transfected BM-MSCs, and these exosomes delivered miR-124 into pancreatic cancer cells, and presented the anti-tumor effects in vitro and in vivo. CONCLUSION: MiR-124-carried BM-MSC-derived exosomes have potential applications for the treatment of pancreatic tumors. METHODS: The expression of miR-124 and EZH2 was determined in both pancreatic cancer tissues and cell lines. miR-124 or EZH2 was overexpressed in AsPC-1 and PANC1 cells. Then, the effects on cell viability. apoptosis, invasion, migration and epithelial mesenchymal transition were evaluated. Afterwards, the roles of miR-124 on the expression and function of EZH2 in pancreatic tumors were determined by dual luciferase reporter assay. Subsequently, miR-124 was transfected to bone marrow mesenchymal stromal cells (BM-MSCs), and the BM-MSCs derived exosomes were isolated and co-cultured with AsPC-1 and PANC1 cells, or injected into pancreatic cancer tumor-bearing mice.

Contrast-Enhanced Ultrasound Assessment of Renal Parenchymal Perfusion in Patients with Atherosclerotic Renal Artery Stenosis to Predict Renal Function Improvement After Revascularization.

BACKGROUND: Identifying patients with atherosclerotic renal artery stenosis (ARAS) who will be improved in renal function after percutaneous transluminal renal artery stenting (PTRAS) is crucial since most patients show no worthwhile benefit of PTRAS. Although the assessment of renal parenchymal perfusion is useful for the identification, few studies predict the renal functional improvement by evaluating the characteristics of renal perfusion. OBJECTIVE: The aim of this study was to assess the renal parenchymal perfusion in ARAS patients with contrast-enhanced ultrasonography (CEUS) and predict the benefits of renal function after PTRAS utilizing time-intensity curve (TIC) parameters. METHODS: Thirty-eight kidneys in 30 ARAS patients received PTRAS in this study. They were divided into moderate stenosis group (n=25) and severe stenosis group (n=13) and mild dysfunction group (n=14) and moderate dysfunction group (n=24) according to the degree of renal stenosis and radioisotope glomerular filtration rate (rGFR). The baseline assessment of renal function and renal parenchymal perfusion were performed for all patients. rGFR was repeated to evaluate the renal outcome at 4 months after PTRAS. The outcome of PTRAS was classified as improved, stable, or deteriorated compared to the baseline. Time-intensity curve (TIC) parameters obtained from CEUS were analyzed to evaluate the predictive accuracy. RESULTS: TIC parameters (AUC and PI) were positively correlated with renal function (r=0.617, 0.663; P<0.05) but weakly and negatively correlated with the stenosis (r=-0.360, -0.435; P<0.05). Baseline rGFR was not accurate in predicting improved renal function after PTRAS (0.670). The accuracy of the combined prediction model of baseline AUC and PI (0.889) was higher than the individual indicators (baseline AUC: 0.855 and PI: 0.782). CONCLUSION: CEUS could accurately assess renal parenchymal perfusion and identify ARAS patients with potential benefit after PTRAS. The combination of TIC parameters (AUC and PI) is valuable in the prediction of improved renal function after PTRAS.

Value of early diagnosis of sepsis complicated with acute kidney injury by renal contrast-enhanced ultrasound.

BACKGROUND: The incidence of acute kidney injury (AKI) in patients with sepsis is high, and the prognosis of patients with septic AKI is poor. The early diagnosis and treatment of septic AKI is of great significance in improving the prognosis of patients with sepsis. AIM: To explore the value of contrast-enhanced ultrasound (CEUS), serum creatinine (Scr), and other indicators in the early diagnosis of septic AKI. METHODS: Ninety patients with sepsis during hospitalization at Tongji Hospital of Tongji University were recruited as subjects. Each patient was recorded with relevant basic data, clinical indicators, and CEUS results. The patients were divided into AKI group and non-AKI group according to the results of renal function diagnosis after 48 h. On the 7th day, the renal function of the non-AKI group was re-evaluated and the patients were further divided into AKI subgroup and non-AKI subgroup. The differences of the indicators in different groups were compared, and the diagnostic value of each indicator and their combination for septic AKI was analyzed. RESULTS: Systemic inflammatory response score (2.58 ± 0.75), blood lactic acid (3.01 ± 1.33 mmol/L), Scr (141.82 ± 27.19 µmol/L), blood urea nitrogen (4.41 ± 0.81mmol/L), and rise time (10.23 ± 2.63 s) in the AKI group were higher than those in the non-AKI group. Peak intensity (PI) (10.78 ± 3.98 dB) and wash in slope (WIS) (1.07 ± 0.53 dB/s) were lower than those in the non-AKI group. The differences were statistically significant (P < 0.05). The PI (12.83 ± 3.77 dB) and WIS (1.22 ± 0.68 dB/s) in the AKI subgroup were lower than those in the non-AKI subgroup, and the differences were statistically significant (P < 0.05). The area under curve (AUC) of Scr for the diagnosis of septic AKI was 0.825 with a sensitivity of 56.76% and a specificity of 100%. The AUCs of WIS and PI (0.928 and 0.912) were higher than those of Scr. Their sensitivities were 100%, but the specificities were 71.70% and 75.47%. The AUC of the combination of three indicators for the diagnosis of septic AKI was 0.943, which was significantly higher than the AUC diagnosed by each single indicator. The sensitivity was 94.59%, and the specificity was 81.13%. CONCLUSION: The combination of Scr, PI, and WIS can improve the diagnostic accuracy of septic AKI. PI and WIS are expected to predict the occurrence of early septic AKI.

Roles and mechanisms of action of HNF­4α in the hepatic differentiation of WB­F344 cells.

Hepatocyte nuclear factor 4 α (HNF­4α) is a nuclear receptor and mediates hepatic genes. WB­F344 liver epithelial cells can differentiate into hepatocytes. The present study aimed to examine the roles and mechanisms of action of HNF­4α on the hepatic differentiation of WB­F344 cells. WB­F344 cells were divided into a normal cell group (WB­F344), empty vector group (PLKO), and gene silencing group (PLKO­SH). The expression levels of HNF­4α were measured using reverse transcription­quantitative polymerase chain reaction analysis. Proliferation of the cells was determined using a Cell Counting kit­8 assay. Based on western blot analysis, the protein levels of α­fetoprotein (AFP), albumin (ALB) and cytokeratin 19 (CK19) were determined. The positive cell rates of the three groups were assessed using periodic acid­Schiff (PAS) staining. Following construction of an RNA­sequencing library, differentially expressed genes (DEGs) between the HNF­4α­silenced and normal samples were screened using the limma package and enrichment analysis was conducted using the DAVID tool. Protein­protein interaction (PPI) and microRNA­targeted regulatory networks were constructed in Cytoscape software. The PLKO­SH group exhibited a lower mRNA level of HNF­4α, higher protein level of AFP, lower protein levels of ALB and CK19, increased cell proliferation, and a lower PAS­positive cell rate. The HNF­4α­silenced and normal samples differed in 499 DEGs. In the PPI network, matrix metallopeptidase 9 (MMP9), early growth response 1 (EGR1), SMAD family member 2 (SMAD2), and RAS­related C3 botulinum substrate 2 (RAC2) were key nodes. HNF­4α may promote the differentiation of WB­F344 cells into hepatocytes by targeting MMP9, EGR1, SMAD2 and RAC2.

Caspase-3 suppresses diethylnitrosamine-induced hepatocyte death, compensatory proliferation and hepatocarcinogenesis through inhibiting p38 activation.

It is critical to understand the molecular mechanisms of hepatocarcinogenesis in order to prevent or treat hepatocellular carcinoma (HCC). The development of HCC is commonly associated with hepatocyte death and compensatory proliferation. However, the role of Caspase-3, a key apoptotic executor, in hepatocarcinogenesis is unknown. In this study, we used Caspase-3-deficient mice to examine the role of Caspase-3 in hepatocarcinogenesis in a chemical (diethylnitrosamine, DEN)-induced HCC model. We found that Caspase-3 deficiency significantly increased DEN-induced HCC. Unexpectedly, Caspase-3 deficiency increased apoptosis induced by DEN and the subsequent compensatory proliferation. Intriguingly, we discovered that Caspase-3 deficiency increased the activation of p38 with and without DEN treatment. Moreover, we demonstrated that TNFα and IL1α stimulated increased activation of p38 in Caspase-3 KO hepatocytes compared with wild-type hepatocytes. Finally, we found that inhibition of p38 by SB202190 abrogated enhanced hepatocyte death, compensatory proliferation and HCC induced by DEN in Caspase-3-deficient mice. Overall, our data suggest that Caspase-3 inhibits chemical-induced hepatocarcinogenesis by suppressing p38 activation and hepatocyte death.

An improved method for increasing the efficiency of gene transfection and transduction.

Transfection and transduction using lentivirus has gained attention in biomedical research. To date, how to reach the maximum transfection and viral transduction efficiency is still challenging. Here we compared the transfection and viral transduction efficiency using commercially available transfection reagents including FuGENE 6, Lipofectamine 2000 and Lipofectamine 3000 in different cell lines and primary cultured cells. Enhanced green fluorescent protein (EGFP) was clearly seen in Eppendorf tubes from harvested cells using Lipofectamine 3000 without using a microscope and UV activation. Strong expression of EGFP was observed in HEK293 cells, mouse primary cortical neurons and human umbilical vein endothelial cells (HUVECs) using confocal microscopy. Western blot showed the strongest EGFP expression using cell lysates from Lipofectamine 3000 transfected HEK293 cells and transduced HUVECs compared with Lipofectamine 2000 or FuGENE 6 reagents. Using Cx43 shRNA lentivirus combined with Lipofectamine 3000 transfection reagent, we can achieve about 90% Cx43 knockdown efficacy in HUVECs. Therefore, our results suggest that a much higher transfection and viral transduction efficiency can be attained by using Lipofectamine 3000 transfection reagent.

TRPM2 promotes the proliferation and invasion of pancreatic ductal adenocarcinoma.

The aim of the present study was to investigate transient receptor potential cation channel subfamily M member 2 (TRPM2), a promising therapeutic target and biomarker for pancreatic ductal adenocarcinoma (PDAC) prognosis, in addition to determining its effects regarding tumor progression and invasion. PDAC is a fatal disease with a poor prognosis, and its associated pathogenic molecular mechanisms remain to be determined. In the present study, combined analysis using genomic and transcriptomic data from two PDAC studies was performed to discover a survival­associated biomarker of PDAC. Survival analysis for genes mutated in ≥10 patients was performed using a Kaplan­Meier curve and tested for significance using a log­rank test. Furthermore, gene­expression correlation analysis was performed to determine the genes with the strongest correlations to TRPM2. In addition, a Cell Counting Kit­8 assay, a scratch wound­healing assay and a Transwell assay were performed in the present study to investigate the proliferative, invasive and metastatic ability of PANC­1 cells in TRPM2­overexpressing and downregulated groups. The mutated TRPM2 gene had a strong negative correlation with patient survival probability compared with the normal control group (P=1.06x10­4). Expression of TRPM2 was strongly correlated with expression of probable phospholipid­transporting ATPase IM, γ­parvin, tudor domain containing 9, Toll­like receptor 7 and Scm­like with four MBT domains protein 2 according to the criterion of a correlation coefficient >0.5. Furthermore, the results of the present study demonstrated that the TRPM2 overexpression in a PDAC cell line (PANC­1) promoted cell proliferation, invasion and metastatic ability. TRPM2 represents a potential therapeutic target and prognostic marker for patients with PDAC. TRPM2 regulates cell proliferation, invasion and migration; however, the underlying mechanism requires further investigation in future studies.