Exosome Marker Proteins of Tumor-Associated Fibroblasts and Exosome-Derived miR-92a-3p Act as Potential Biomarkers for Liver Cancer.

Many recent studies have shown that microRNAs (miRNAs) in exosomes can be absorbed by nearby or distant cells, and the abnormal expression of these exosomal miRNAs is associated with most pathological progresses. In this study, we explored the diagnostic value of exosome marker proteins and exosome-derived miR-92a-3p in liver cancer. The clinicopathological data of 60 patients with liver cancer admitted to Tanghan Gongren Hospital from October 2017 to October 2019 were collected. Tumor tissue and adjacent tissue were collected during surgery. Quantitative reverse transcription polymerase chain reaction and Western blot were used to detect the expression levels of miR-92a-3p in exosomes of fibroblasts and tumor tissue, and exosome marker proteins. In liver cancer tissue and fibroblast exosomes, the expression of miR-92a-3p was significantly increased. The receiver operator characteristic curve of the expression level of miR-92a-3p in exosomes and tissue showed that the area under the curve was 0.906 and 0.911, respectively. HSP70 and CD63 were highly expressed in the tissue of liver cancer and fibroblast exosomes. miR-92a-3p was positively correlated with HSP70 and CD63 in the exosomes of liver cancer fibroblasts. In addition, miR-92a-3p and exosome marker proteins (HSP70 and CD63) were highly expressed in tumors with a diameter greater than 5 cm, and were higher in liver cancer patients with BCLC stage B/C. Tumor fibroblast-derived exosome marker proteins and miR-92a-3p have good diagnostic value in liver cancer, indicating that they may be new diagnostic markers for liver cancer.

Identification and construction of a 13-gene risk model for prognosis prediction in hepatocellular carcinoma patients.

We attempted to screen out the feature genes associated with the prognosis of hepatocellular carcinoma (HCC) patients through bioinformatics methods, to generate a risk model to predict the survival rate of patients. Gene expression information of HCC was accessed from GEO database, and differentially expressed genes (DEGs) were obtained through the joint analysis of multi-chip. Functional and pathway enrichment analyses of DEGs indicated that the enrichment was mainly displayed in biological processes such as nuclear division. Based on TCGA-LIHC data set, univariate, LASSO, and multivariate Cox regression analyses were conducted on the DEGs. Then, 13 feature genes were screened for the risk model. Also, the hub genes were examined in our collected clinical samples and GEPIA database. The performance of the risk model was validated by Kaplan-Meier survival analysis and receiver operation characteristic (ROC) curves. While its universality was verified in GSE76427 and ICGC (LIRI-JP) validation cohorts. Besides, through combining patients' clinical features (age, gender, T staging, and stage) and risk scores, univariate and multivariate Cox regression analyses revealed that the risk score was an effective independent prognostic factor. Finally, a nomogram was implemented for 3-year and 5-year overall survival prediction of patients. Our findings aid precision prediction for prognosis of HCC patients.

miR-9-5p facilitates hepatocellular carcinoma cell proliferation, migration and invasion by targeting ESR1.

The study aimed to explore the relationship between miR-9-5p and ESR1, and clarify the underlying functional mechanism in the occurrence and development of hepatocellular carcinoma (HCC). Expression data including miRNAs and mRNAs of HCC downloaded from TCGA database were processed for differential analysis, and corresponding clinical data were collected for survival analysis to identify the target miRNA miR-9-5p. Bioinformatics databases were applied for predicting downstream target mRNAs of miR-9-5p. qRT-PCR was used to evaluate expression of miR-9-5p. Western blot was used to detect protein expression of ESR1. MTT, wound healing assay and Transwell assay were used to detect HCC cell proliferation, migration and invasion, respectively. Dual-luciferase reporter gene assay was used to identify the targeting relationship between miR-9-5p and ESR1. Research suggested that miR-9-5p was highly expressed in HCC cells but ESR1 was poorly expressed. Overexpression of miR-9-5p could improve the proliferation, invasion and migration of cells. Dual-luciferase reporter assay showed that ESR1 was the downstream target of miR-9-5p in HCC. Overexpression of miR-9-5p markedly reduced ESR1 mRNA and protein levels in HCC cells, whereas inhibition of miR-9-5p expression produced the contrary results. Silencing ESR1 could noticeably reverse the effect of miR-9-5p knockdown on the proliferation, migration and invasion of HCC cells. As an oncogene, miR-9-5p fostered the proliferation, migration and invasion of HCC cells by targeting and inhibiting ESR1 expression.

Hydrogen sulphide exacerbates acute pancreatitis by over-activating autophagy via AMPK/mTOR pathway.

Previously, we have shown that hydrogen sulphide (H2 S) might be pro-inflammatory during acute pancreatitis (AP) through inhibiting apoptosis and subsequently favouring a predominance of necrosis over apoptosis. In this study, we sought to investigate the detrimental effects of H2 S during AP specifically with regard to its regulation on the impaired autophagy. The incubated levels of H2 S were artificially intervened by an administration of sodium hydrosulphide (NaHS) or DL-propargylglycine (PAG) after AP induction. Accumulation of autophagic vacuoles and pre-mature activation of trypsinogen within acini, which indicate the impairment of autophagy during AP, were both exacerbated by treatment with NaHS but attenuated by treatment with PAG. The regulation that H2 S exerted on the impaired autophagy during AP was further attributed to over-activation of autophagy rather than hampered autophagosome-lysosome fusion. To elucidate the molecular mechanism that underlies H2 S-mediated over-activation of autophagy during AP, we evaluated phosphorylations of AMP-activated protein kinase (AMPK), AKT and mammalian target of rapamycin (mTOR). Furthermore, Compound C (CC) was introduced to determine the involvement of mTOR signalling by evaluating phosphorylations of downstream effecters including p70 S6 kinase (P70S6k) and UNC-51-Like kinase 1 (ULK1). Our findings suggested that H2 S exacerbated taurocholate-induced AP by over-activating autophagy via activation of AMPK and subsequently, inhibition of mTOR. Thus, an active suppression of H2 S to restore over-activated autophagy might be a promising therapeutic approach against AP-related injuries.

Necroptosis: a potential, promising target and switch in acute pancreatitis.

Pancreatic acinar cell death is the major pathophysiological change in early acute pancreatitis (AP), and the death modalities are important factors determining its progression and prognosis. During AP, acinar cells undergo two major modes of death, including necrosis and apoptosis. Acinar necrosis can lead to intensely local and systemic inflammatory responses, which both induce and aggravate the lesion. Necrosis has long been considered an unregulated, and passive cell death process. Since the effective interventions of necrosis are difficult to perform, its relevant studies have not received adequate attention. Necroptosis is a newly discovered cell death modality characterized by both necrosis and apoptosis, i.e., it is actively regulated by special genes, while has the typical morphological features of necrosis. Currently, necroptosis is gradually becoming an important topic in the fields of inflammatory diseases. The preliminary results from necroptosis in AP have confirmed the existence of acinar cell necroptosis, which may be a potential target for effectively regulating inflammatory injuries and improving its outcomes; however, the functional changes and mechanisms of necroptosis still require further investigation. This article reviewed the progress of necroptosis in AP to provide a reference for deeply understanding the pathogenic mechanisms of AP and identifying new therapeutic targets.

Acinar cell cystadenoma of the pancreas: A retrospective analysis of ten-year experience from a single academic institution.

BACKGROUND/OBJECTIVE: Acinar cell cystadenoma (ACA), also referred to as "acinar cystic transformation", is a rare and newly recognized cystic lesion of the pancreas displaying apparent acinar cell differentiation with benign outcomes. We summarized our experience with the diagnosis, clinicopathologic feature, treatment and prognosis of ACA to provide a reference for the disposal of this uncommon condition. METHODS: We retrospectively analyzed the clinical data from eight patients with ACA treated in our hospital between March, 2005 and January, 2015. RESULTS: Among eight patients, five of whom were female and the age at diagnosis ranged from 33 to 67 years (mean, 49.8 y). The most commonly clinical symptom was abdominal pain. Eight lesions were unifocal and either unilocular (n = 5) or multilocular (n = 3) with average size of 10.5 cm (range, 5.1-19.7 cm). All the patients were treated surgically and a definite diagnosis of ACA was obtained by the histopathological, histochemical and immunohistochemical tests. The length of stay range was from 11 to 17 days and there were no perioperative deaths. At a median follow-up of 57.3 months, all the patients were alive and there was no evidence of recurrence, distant metastasis or malignant transformation. CONCLUSIONS: Appropriately preoperative differential diagnosis of ACA remains challenging and the final result is usually gained by the histopathology and immunohistochemistry. Although the origin of ACA is still contradictory, surgery is actively advocated as the most effective method for relieving the symptoms and preventing the tumor from local extension or malignant transformation so as to obtain an optimal long-term survival.

m5C RNA methyltransferase-related gene NSUN4 stimulates malignant progression of hepatocellular carcinoma and can be a prognostic marker.

Hepatocellular carcinoma (HCC) is a cancer with relatively high mortality, yet little attention has been devoted for related prognostic biomarkers. This study analyzed differential expression of m5C RNA methyltransferase-related genes in normal samples and tumors samples in TCGA-LIHC using Wilcoxon test. K-means consensus clustering analysis was implemented to subdivide samples. Independent prognostic factors were screened by univariate and multivariate Cox regression analyses. KEGG pathway enrichment analysis was performed on the screened independent prognostic factor using GSEA tools. qPCR was conducted to test mRNA expression of key m5C RNA methyltransferase-related genes in tissues and cells. There were 7 m5C RNA methyltransferase-related genes (NOP2, NSUN4, etc.) differentially expressed in HCC tumor tissues. HCC samples were classified into 3 subgroups through clustering analysis according to the expression mode of m5C RNA methyltransferase-related genes. It was also discovered that patients in different subgroups presented significant differences in survival rate and distribution of grade. Additionally, NOP2, NSUN4 and NSUN5 expression notable varied in different grades. Through regression analyses combined with various clinical pathological factors, it was displayed that NSUN4 could work as an independent prognostic factor. KEGG analysis showed that NSUN4 mainly enriched in signaling pathways involved in ADHERENS JUNCTION, RNA DEGRADATION, MTOR SIGNALING PATHWAY, COMPLEMENT and COAGULATION CASCADES. As examined by qPCR, NSUN4 was conspicuously upregulated in HCC patient's tissues and cells. Altogether, our study preliminarily developed a novel biomarker that could be independently used in prognosis of HCC, which may provide a new direction for the study of related molecular mechanism or treatment regimen.

MiR-18a-5p Facilitates Progression of Hepatocellular Carcinoma by Targeting CPEB3.

Objective: To explore the function of the miR-18a-5p/CPEB3 axis in regulating the occurrence of hepatocellular carcinoma (HCC). Methods: Differentially expressed miRNAs and mRNAs were acquired by bioinformatics analysis. qRT-PCR was used for miR-18a-5p and CPEB3 mRNA expression detection. Cell functional assays were implemented to examine the biological functions of HCC cells. The binding relationship between miR-18a-5p and CPEB3 was verified by a dual luciferase assay. Results: In HCC, miR-18a-5p was remarkably highly expressed, while CPEB3 was markedly lowly expressed. HCC cell progression was facilitated after cells transfecting miR-18a-5p mimic, whereas silencing miR-18a-5p caused the opposite result. Overexpressing CPEB3 could restore promoting effect of miR-18a-5p on the growth of HCC cells. Conclusion: Oncogene miR-18a-5p accelerates malignant phenotype by suppressing CPEB3. MiR-18a-5p/CPEB3 axis in HCC identified in this study provides a new target for HCC treatment.

mir-182-5p Regulates Cell Growth of Liver Cancer via Targeting RCAN1.

Regulator of calcineurin 1 (RCAN1) is an endogenous protein that is involved in the regulation of the occurrence and progression of a variety of cancers, but currently, people know little about its potential mechanism. This study investigated the function and mechanism of RCAN1 and miR-182-5p in liver cancer cells. In this study, reliable data demonstrated that RCAN1 suppressed cell proliferation, migration, invasion, and cell cycle progression of liver cancer. Additionally, the expression of RCAN1 was noted to be regulated by its upstream regulator miR-182-5p, and miR-182-5p was prominently highly expressed in liver cancer cells. Based on this, it was further proved through cell experiments that miR-182-5p facilitated cell growth of liver cancer through RCAN1 downregulation, showing that RCAN1 may be a fresh biomarker and target for diagnosis and treatment of liver cancer.

MiR-92a-3p Promotes the Malignant Progression of Hepatocellular Carcinoma by Mediating the PI3K/AKT/mTOR Signaling Pathway.

BACKGROUND: As one of the most common cancers globally, hepatocellular carcinoma (HCC) usually has a poor prognosis. Many HCC patients are usually diagnosed at advanced stages. Therefore, new potential biomarkers for the diagnosis and prognosis of HCC are urgently needed. More and more studies have shown that miR-92a-3p can regulate the occurrence and development of various cancers, but its clinical significance and molecular mechanism in HCC are still elusive. Here, we tried to clarify the regulatory mechanism of miR-92a-3p in HCC. METHODS: In this study, we conducted qRT-PCR and revealed that miR-92a-3p was notably upregulated in HCC cells. MTT, flow cytometry, wound healing, Transwell invasion assays and western blot were conducted to uncover that overexpressed miR-92a-3p could boost the proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of HCC cells while inhibiting cell apoptosis. In addition, the proteins associated with the PI3K/AKT/mTOR pathway were also detected by western blot. RESULTS: It was suggested that miR-92a-3p could activate the PI3K/AKT/mTOR signaling pathway. CONCLUSION: These results suggest that miR-92a-3p plays a tumor-promoting role in HCC and may be a potential biomarker for the diagnosis and prognosis of HCC.

MiR-452-5p mediates the proliferation, migration and invasion of hepatocellular carcinoma cells via targeting COLEC10.

Objective: This study explored the potential function of miR-452-5p in hepatocellular carcinoma (HCC) and clarified the mechanism underlying HCC progression. Materials & methods: Real-time quantitative PCR was used to detect miR-452-5p and COLEC10 mRNA expression in HCC, western blot was performed to test COLEC10 protein expression. The regulatory mechanism of miR-452-5p/COLEC10 in HCC cells was explored using CCK-8, wound healing assay, Transwell and dual-luciferase reporter assay. Results: MiR-452-5p was greatly upregulated in HCC cells, and it served as an oncogene playing an active role in HCC cell proliferation, migration and invasion. COLEC10 was identified as the target of miR-452-5p in HCC attenuating the promoting effect of miR-452-5p on HCC cells upon overexpression. Conclusion: MiR-452-5p can promote the progression of HCC via targeting COLEC10.

Inhibition of RIPK1-dependent regulated acinar cell necrosis provides protection against acute pancreatitis via the RIPK1/NF-κB/AQP8 pathway.

Currently, preliminary results have confirmed the existence of receptor-interacting protein kinase 3 (RIPK3) and mixed lineage kinase domain-like protein (MLKL)-dependent necroptosis of pancreatic acinar cells during early acute pancreatitis (AP), which might be a potential target for the effective regulation of necroinflammatory injury. However, the exact effect of receptor-interacting protein kinase 1 (RIPK1)-dependent regulated acinar cell necrosis on AP is still uncertain. In our study, we first explored the changes in the degree of local and systemic inflammation in AP rats when the activation of acinar cell RIPK1 was inhibited. The RIPK1 inhibitor Nec-1 was used to treat rats, and the levels of related inflammatory markers, necrosis indicators and apoptotic indicators were measured. Changes in pancreatic nuclear factor κB (NF-κB) and aquaporin 8 (AQP8) expression were noted. Next, the expression of AQP8 in AR42J cells was inhibited, and the degree of cell necrosis and inflammatory damage was found to be significantly reduced. Most importantly, we demonstrated that the RIPK1/NF-ĸB/AQP8 axis might be a potential regulatory pathway mediating RIPK1-dependent regulated acinar cell necrosis in early AP. Finally, we used the NF-κB inhibitor PDTC and Nec-1 to treat rats in different groups and measured the degree of pathological pancreatic injury, the activation of RIPK1, and the expression of NF-κB and AQP8. In summary, we hypothesized that there might be a RIPK1/NF-ĸB/AQP8 pathway controlling RIPK1-dependent regulated necrosis of acinar cells in AP, which might be a promising therapeutic target against AP-related injury.

Acute pancreatitis induced by etoposide-lobaplatin combination chemotherapy used for the treatment of lung cancer: A case report and literature review.

RATIONALE: Drug-induced pancreatitis (DIP) is a rare type of pancreatitis that is not usually observed in the clinical practice. It is generally difficult to distinguish from acute pancreatitis (AP) induced by other causes. PATIENT CONCERNS: Here, we report a 62-year-old Chinese female patient with "small cell lung cancer" as the initial presentation. Because the patient could not bear the surgical treatment, the chemotherapy composed of lobaplatin and etoposide was performed. Three days later, the patient displayed sudden abdominal pain, distension, nausea, and vomiting without obvious inducements. Laboratory tests showed that the levels of serum and urine amylase were enhanced; abdominal computed tomography (CT) result showed the enlargement of the pancreas, peripancreatic effusion, and a rough edge, which suggested the diagnosis of AP. The patient had no history of biliary tract disease, alcoholism, binge overeating, hyperlipidemia, and hereditary pancreatitis. DIAGNOSES: The patient was diagnosed with DIP. INTERVENTIONS: The chemotherapy was stopped at once and we performed fluid resuscitation, pain alleviation, prophylactic antibiotics, and nutritional support, etc on the patient. Later, the patient's clinical symptoms were obviously relieved, and she recovered successfully. OUTCOMES: The chemotherapy was continued, but later, the patient showed abdominal pain, distension, nausea, and vomiting again. The levels of serum amylase and urine amylase were enhanced again. Further imaging examination strongly indicated the recurrence of AP. LESSONS: We should raise awareness of the clinicians regarding DIP, thereby enabling its timely diagnosis and accurate treatment, as well as promoting the rational and safe use of drugs.

Anatomic Characteristics, Identification, and Protection of the Nonrecurrent Laryngeal Nerve during Thyroidectomy.

Objective We aimed to investigate the anatomical features and variation pattern of the nonrecurrent laryngeal nerve (NRLN), summarize the methods for identifying the NRLN before and during thyroidectomy, and share experiences regarding preventing and treating its injury. Study Design Retrospective case data analysis. Setting First Affiliated Hospital of Harbin Medical University. Subjects and Methods Between January 2002 and May 2016, 7392 patients underwent thyroidectomy in our hospital. Of them, 28 patients with NRLN were identified, and their clinical data were retrospectively analyzed. Results This study included 7392 patients in which the recurrent laryngeal nerves (RLNs) were routinely identified during surgery. The presence of NRLN was intraoperatively confirmed in 28 patients. All the NRLNs were located on the right side and its overall incidence was 0.37%. Five of the NRLNs were classified as type I, 19 as type IIa, and 4 as type IIb. Of the 28 cases, 4 NRLNs were injured during surgery, in which primary end-to-end anastomosis or local seal with corticosteroid injection was performed as a remedy. In the 4 patients with NRLN injury, 2 presented with postoperative hoarseness that indicated vocal cord paralysis confirmed by laryngoscope; the other 2 patients' voices had no significant changes. Conclusion The NRLN, which is rare in clinical practice and predominantly right-sided, is anatomically more complex and variant at a higher risk of surgical injury. The key factors to accurately identify NRLN and to effectively prevent its injury include careful interpretation of auxiliary examination results before surgery, raising awareness of its presence, meticulous dissection, and routine exposure of the RLN during surgery.

External Versus Internal Pancreatic Duct Drainage for the Early Efficacy After Pancreaticoduodenectomy: A Retrospectively Comparative Study.

PURPOSE: To compare the early efficacy of external versus internal pancreatic duct drainage after pancreaticoduodenectomy (PD), providing clinical evidence for selecting the optimal approach to pancreatic duct drainage. MATERIAL AND METHODS: The clinical data of 395 consecutive patients undergoing PD from 2006 to 2013 were analyzed retrospectively. All the patients were divided into external and internal drainage group. Intraoperative blood loss, surgery duration, postoperative hospitalization duration, mortality rate, PF, and other complications were compared between the two groups. The perioperative relative risk factors that might induce PF were analyzed. RESULTS: External drainage significantly reduced the incidences of post-PD PF, delayed gastric emptying, abdominal infection, bowel obstruction, overall complications, and shortened the healing time of PF (p < .05). The univariate analysis showed that the pancreatic duct drainage method, body mass index (BMI), preoperative serum bilirubin level, perioperative blood transfusion, pancreaticojejunostomy approach, pancreatic texture, pancreatic duct diameter, and primary disease differed markedly between the two groups (p < .05). A multivariate analysis revealed that BMI ≥ 25 kg/m(2), internal pancreatic duct drainage, pancreatic duct diameter <3 mm, soft pancreatic texture, and ampullary disease were independent risk factors for PF. CONCLUSIONS: External pancreatic duct drainage can effectively reduce the morbidity of PF and overall complications after PD.