Exosomes secreted by M2 macrophages promote cancer stemness of hepatocellular carcinoma via the miR-27a-3p/TXNIP pathways.

OBJECTIVE: Accumulating evidence has suggested that microRNAs (miRNAs) derived from M2 macrophage-derived exosomes (M2 exosomes) can regulate the progression of hepatocellular carcinoma (HCC). Nevertheless, the effect of miR-27a-3p derived from M2 exosomes on HCC has not been reported. We aim to explore the role of M2 exosomal miR-27a-3p in the cancer stemness of HCC via regulating thioredoxin-interacting protein (TXNIP). METHODS: Exosomes were extracted from transfected M2 macrophages and were then co-cultured with HCC cells. Expression of miR-27a-3p and TXNIP, stemness, proliferation, drug resistance, migration, invasion and in vivo tumorigenicity of HCC cells were determined to assess the role of M2 exosomal miR-27a-3p in HCC. The binding relationship between miR-27a-3p and TXNIP was detected. RESULTS: MiR-27a-3p was upregulated and TXNIP was downregulated in HCC cells, and M2 exosomes further upregulated miR-27a-3p. The upregulated M2 exosomal miR-27a-3p promoted stemness, proliferation, drug resistance, migration, invasion and in vivo tumorigenicity of HCC cells. TXNIP was confirmed as a target gene of miR-27a-3p. CONCLUSION: M2 macrophages-derived exosomal miR-27a-3p promotes cancer stemness of HCC via downregulating TXNIP.

Downregulation of serum exosomal miR-320d predicts poor prognosis in hepatocellular carcinoma.

BACKGROUND: MicroRNAs (miRNAs) is a class of functional regulator of tumorigenesis of human cancer including hepatocellular carcinoma (HCC). However, the potential clinical significance of serum exosomal miR-320d in HCC has not been elucidated. METHODS: Real-time reverse transcription PCR was used to detect the expression pattern of serum exosomal miR-320d in patients with HCC, and the correlation between the deregulation of serum exosomal miR-320d and the clinical outcome of HCC was explored. The biological function of exosomal miR-320d in HCC was also investigated. RESULTS: Our results showed that the expression levels of exosomal miR-320d were remarkably reduced in the serum samples of HCC patients and the culture medium of HCC cell lines compared with their respective controls. Serum exosomal miR-320d could differentiate the HCC patients from healthy controls with high accuracy. In addition, its level was remarkably increased in the HCC patients who had received surgical treatment. Moreover, reduced serum exosomal miR-320d was associated with advanced tumor stage, positive lymph node metastasis, and poorly differentiated tumors. HCC patients with lower serum exosomal miR-320d had shorter overall and disease-free survival. Low serum exosomal miR-320d was identified to be an independent unfavorable prognostic factor for HCC. Finally, overexpression of miR-320d inhibited the proliferation and invasion of HCC cells, and BMI1 was demonstrated to be a direct target of miR-320d. CONCLUSION: Taken together, serum exosomal miR-320d could be a potential non-invasive biomarker for the diagnosis and prognosis of HCC.

Correlation between the glucose level and the development of acute pancreatitis.

OBJECTIVE: To investigate the correlation between the level of glucose in serum and the development of acute pancreatitis (AP). METHODS: Data of 153 AP cases were collected, in which there were 130 patients with mild AP (MAP), 4 with moderate-severe AP (MSAP) and 19 with severe AP (SAP). At the time of admission, following indexes of patients were recorded: glucose, APACHE II score, TNF-α and C-reaction protein (CRP). RESULTS: At the time of admission, the levels of glucose in serum and APACHE II scores in the MSAP and SAP groups were significantly higher than those in the MAP group, but after treatment, the level of glucose in serum was recovered in 95.8% of the patients in the MAP group, while this digit in the SAP group remained to be 68.4%; in the SAP group, the levels of TNF-α and CRP in patients with sustained hypertension were significantly higher than those with non-persistent hypertension; in terms of the length of stay in hospital, the SAP group was shorter than that in the non-treatment group, and the difference had statistical significance (p < 0.05). Moreover, we found that the level of glucose in serum was positively correlated with the APACHE II scores, TNF-α and CRP. CONCLUSION: Glucose level in serum can be used as one of the indicators for evaluating the severity and development of AP in clinical practice.

LncRNA NEAT1-MicroRNA-140 axis exacerbates nonalcoholic fatty liver through interrupting AMPK/SREBP-1 signaling.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is a severe liver disease, which influences the health of people worldwide. However, the mechanism modulating the pathogenesis of NAFLD remains elusive. It was reported that nuclear enriched abundant transcript 1 (NEAT1) and microRNA-140 (miR-140) could regulate lipogenesis, but whether they could influence NAFLD are still unknown. METHODS: HepG2 cells were treated by free fatty acids (FFA) to establish the model of NAFLD in vitro, and C57 mice were treated by high-fat diet to establish the model of NAFLD in vivo. Cell transfection was applied to regulate the expression of NEAT1 and miR-140. Western blotting and qRT-PCR were applied for measuring expression of protein and mRNA, respectively. HE staining and Oil Red O staining were used for observing liver tissues. RESULTS: NEAT1 and miR-140 are upregulated in hepacytes under the NAFLD conditions. NEAT1 directly binds to miR-140 and acts synergistically with miR-140 to exacerbate the progression of NAFLD. Reciprocally, silence of miR-140 or NEAT1 alleviates the severity of NAFLD. The mechanistical study shows that the axis of NEAT1-miR-140 inactivates AMPK/SREBP-1 signaling during the NAFLD. . CONCLUSION: The NEAT1-miR-140 axis play a crucial role in modulation of NAFLD via inactivation of AMPK/SREBP1 signaling. This study may provide a novel insight for the treatment of NAFLD.

Vibrio vulnificus VvhA induces Th1 and Tfh cells to proliferate against Vibrio vulnificus in a mouse model of infection.

AIM: To characterize the roles of VvhA in host's acquired immune response to Vibrio vulnificus infection. MATERIALS & METHODS: The recombinant VvhA fusion protein was used to immunize mice and the anti-VvhA polyclonal antibody was produced in vivo for prophylactic and therapeutic efficacy assay. The roles of VvhA in T helper (Th) cells differentiation were analyzed by vvhA-deleted mutant during the early phase of infection, while the ratio of Th2 and T follicular helper (Tfh) cells were examined in VvhA immunization. RESULTS: Anti-VvhA antibody exhibited neutralization activity against V. vulnificus. Wild-type strain induced higher level of Th1 cells than the mutant, and the concentrations of IgG2a and IFN-γ were increased during the early phase of infection. The spontaneous development of Tfh was observed in immunized model, and the serum IL-21 was increased. CONCLUSION: V. vulnificus VvhA elicited cellular and humoral immune responses by Th1 and Tfh cells to provide protection against VvhA.

Helicobacter pylori with East Asian-type cagPAI genes is more virulent than strains with Western-type in some cagPAI genes

Abstract The severity of Helicobacter pylori-related disease is correlated with the presence and integrity of a cag pathogenicity island (cagPAI). cagPAI genotype may have a modifying effect on the pathogenic potential of the infecting strain. After analyzing the sequences of cagPAI genes, some strains with the East Asian-type cagPAI genes were selected for further analysis to examine the association between the diversity of the cagPAI genes and the virulence of H. pylori. The results showed that gastric mucosal inflammatory cell infiltration was significantly higher in patients with East Asian-type cagPAI genes H. pylori strain compared with mosaicism cagPAI genes H. pylori strain (p < 0.05). H. pylori strains with the East Asian-type cagPAI genes were closely associated with IL-8 secretion in vitro and in vivo compared with H. pylori strains with the mosaicism cagPAI genes (p < 0.01). H. pylori strains with East Asian-type cagPAI genes are able to strongly translocate CagA to host cells. These results suggest that H. pylori strains with East Asian-type cagPAI genes are more virulent than the strains of cagPAI gene/genes that are Western type.

Helicobacter pylori with East Asian-type cagPAI genes is more virulent than strains with Western-type in some cagPAI genes.

The severity of Helicobacter pylori-related disease is correlated with the presence and integrity of a cag pathogenicity island (cagPAI). cagPAI genotype may have a modifying effect on the pathogenic potential of the infecting strain. After analyzing the sequences of cagPAI genes, some strains with the East Asian-type cagPAI genes were selected for further analysis to examine the association between the diversity of the cagPAI genes and the virulence of H. pylori. The results showed that gastric mucosal inflammatory cell infiltration was significantly higher in patients with East Asian-type cagPAI genes H. pylori strain compared with mosaicism cagPAI genes H. pylori strain (p<0.05). H. pylori strains with the East Asian-type cagPAI genes were closely associated with IL-8 secretion in vitro and in vivo compared with H. pylori strains with the mosaicism cagPAI genes (p<0.01). H. pylori strains with East Asian-type cagPAI genes are able to strongly translocate CagA to host cells. These results suggest that H. pylori strains with East Asian-type cagPAI genes are more virulent than the strains of cagPAI gene/genes that are Western type.

Concurrent Percutaneous Transhepatic Papillary Balloon Dilatation Combined with Laparoscopic Cholecystectomy for the Treatment of Gallstones with Common Bile Duct Stones.

OBJECTIVE: To evaluate the clinical effectiveness and safety of percutaneous transhepatic papillary balloon dilatation (PPBD) combined with laparoscopic cholecystectomy (LC) for the treatment of gallstones with common bile duct stones. MATERIALS AND METHODS: From February 2012 to January 2013, 65 patients (31 males and 34 females) who had gallstones with common bile duct stones were divided into two groups: endoscopic papillary balloon dilatation (EPBD)+LC (n = 40 cases) and PPBD+LC (n = 25 cases). The maximum transverse diameter of common bile duct stoned was 15 mm. White blood cell count and serum amylase level were measured at 12 hours and 72 hours after the operation, and operation time, time of gastrointestinal function recovery, and hospitalization time were analyzed. RESULTS: No case in either group was converted to open surgery. The stones were successfully pushed out into the duodenum in all patients of both groups. One case of postoperative pancreatitis (2.5%) and 5 cases of hyperamylasemia (12.5%) occurred among the 40 patients in the EPBD+LC group. In contrast, no case of postoperative pancreatitis and 2 cases of hyperamylasemia (8%) occurred among the 25 patients in the PPBD+LC group. The serum amylase level at 12 hours postoperatively was statistically significantly different (307 ± 39.94 IU/L in the EPBD+LC group and 193 ± 30.78 IU/L in the PPBD+LC group; P < .05). There were no significant differences between the two groups in operative time, postoperative gastrointestinal function recovery time, or hospital stay time (P > .05). During follow-up of 2 years, 1 case of recurrence for common bile duct stones was discovered and was treated successfully with EPBD repeatedly. CONCLUSIONS: PPBD combined with LC simultaneously is an alternative effective and safe procedure for the treatment of gallstones and bile duct stones, especially for those patients who are unsuitable for endoscopic treatment.

[Clinical study of trans-T tube duodenal papilla balloon dilatation: a novel procedure for postsurgical residual common bile duct stone].

OBJECTIVE: To investigate the safety and efficacy of trans-T tube duodenal papilla balloon dilatation for the removal of residual common bile duct stone after choledocholithotomy and T tube drainage. METHODS: Thirteen cases with residual common bile duct stone treated with trans-T tube duodenal papilla balloon dilatation in our department from June 2010 to April 2012 were analyzed retrospectively. Record CA19-9, bilirubin and albumin before the procedure, 1 week and 1 month later. Check if immediate complications occurs, including hemorrhage, perforation, pancreatitis and cholangitis. During the following up for 2 years, stone recurrence, reflux cholangitis and other long-term complications were observed. Analyze the changes of indicators between preoperative and postoperative and the correlation. RESULTS: All of the 13 patients were treated successfully. 11 patients underwent one procedure, and 2 patients received twice or more times of procedures. CA19-9 decreased significantly 1 week later and 1 month later compared with those before the procedure, while ALB was opposite. Incidences of biliary tract infection and hemorrhage were 1 and 1 respectively. No severe complications occurred, including perforation of gastrointestinal or biliary tract. Incidences of recurrent stone and reflux cholangeitis were 2 and 1 in two years after the procedure. CONCLUSIONS: Trans-T tube duodenal papilla balloon dilatation is a safe and effective procedure for patients with recurrent common bile duct stone after choledocholithotomy and T-tube drainage. For the cases which could not be handled with endoscopic procedures, it provides a new therapeutic approach.

[Clinical study of percutaneous transhepatic balloon dilation:a novel procedure for common bile duct stone].

OBJECTIVE: To investigate the safety and efficacy of percutaneous transhepatic balloon dilation for the removal of common bile duct stone. METHODS: Sixty-eight cases with common bile duct stone treated with percutaneous transhepatic balloon dilation in our department from July 2008 to April 2011 were analyzed retrospectively.Record CA19-9, total bilirubin, indirect bilirubin and albumin before the procedure, 1 week and 1 month later. Check if immediate complications occurs, including hemorrhage, perforation, pancreatitis and cholangitis. During the following up for 2 years, stone recurrence, reflux cholangitis and other long-term complications were observed. Analyze the changes of indicators between preoperative and postoperative and the correlation. RESULTS: All of the 68 patients were treated successfully.53 patients underwent one procedure, and 15 patients received twice or more times of procedures. CA19-9, TBIL and IBIL decreased significantly 1 week later and 1 month later compared with those before the procedure, while ALB was opposite.Incidences of biliary tract infection, hemorrhage, and pancreatitis were 4.4% (3/68), 2.9% (2/68) and 1.5% (1/68) respectively.No severe complications occurred, including perforation of gastrointestinal or biliary tract.Incidences of recurrent stone and reflux cholangitis were 14.7% (10/68) and 5.9% (4/68) in two years after the procedure. CONCLUSION: Percutaneous transhepatic balloon dilation is a safe and effective procedure for patients with common bile duct stone. For the cases which could not be handled with endoscopic procedures, it provides a new therapeutic approach.

The role of leptin receptor gene polymorphisms in determining the susceptibility and prognosis of NSCLC in Chinese patients.

AIM: Although the role of genetic polymorphisms of leptin receptor (LEPR) gene in several cancers has been documented, the association between polymorphisms of LEPR gene and lung cancer remains unknown. METHOD: We recruited 744 patients histologically diagnosed as non-small cell lung cancer (NSCLC) and 832 controls in this study. Polymorphism analysis of LEPR gene was performed by PCR-restriction fragment length polymorphisms. RESULTS: The Arg/Arg genotype and Arg allele frequency of the Gln223Arg in LEPR gene were significantly prevalent in NSCLC subjects than in controls (P < 0.05). The odd ratio (OR) for NSCLC in Arg/Arg genotype carriers was 3.12 (95% CI: 2.25-4.56, P = 0.0023, with Gln/Gln as reference). There were no significant differences in the genotype distributions and allele frequencies of Lys109Arg and Lys656Asn in LEPR gene between NSCLC cases and controls (All P > 0.05). The Arg/Arg carriers had higher cancer grade and higher TNM stage. Kaplan-Mier curve showed the Arg/Arg carriers had a poor prognosis than those with Gln/Arg and Gln/Gln genotype carriers. Cox proportional hazards regression models showed the hazard ratio (HR) for death associated with Arg/Arg genotype was 3.43 (95% CI: 2.45-5.92, compared with Gln/Gln carriers, P = 0.002). The other two SNPs of LEPR gene did not show this trend in the evaluation of their role in determining the prognosis of NSCLC subjects. CONCLUSION: The results suggest the polymorphisms of Gln223Arg, rather than Lys109Arg and Lys656Asn, may be used as a molecular marker for progression and prognosis of NSCLC.

[Treatment analysis of 26 patients with breast ductal carcinoma in situ].

OBJECTIVE: To study the appropriate surgical treatment for breast ductal carcinoma in situ (DCIS). METHODS: Twenty-six such patients treated between 1992 and 2001 were retrospectively analyzed. Among them, 3 patients were treated by simple mastectomy, 23 patients by mastectomy and axillary lymph node dissection, 8 patients by chemotherapy and one patient by radiotherapy after operation. Median follow-up was 42 m (rang 12 - 112 m). RESULTS: Except 3 of these 26 patients lost in follow-up and 1 patient died from diabetes mellitus, all the other 22 patients survived over 5 years. All lymph nodes dissected from 23 patients were negative. After surgery, 3 patients developed lymph edema of the arm. CONCLUSION: DCIS, lacking the potential of metastasis, is not invasive. Conservative breast surgery without lymph node dissection is feasible for most DCIS patients.