Silencing of microRNA-375 affects immune function in mice with liver failure by upregulating astrocyte elevated gene-1 through reducing apoptosis of Kupffer cells.

This study aims to investigate how microRNA-375 (miR-375) improves immune function by regulating liver macrophages (Kupffer cells) in mice with liver failure. Forty mice were divided into ConA-1h, ConA-3h, ConA-6h, and control groups, with 10 mice in each group. Mice models of liver failure were established by injecting concanavalin A (ConA) solution via the tail veins of mice, and then primary Kupffer cells were isolated and cultured. Reverse transcription quantitative polymerase chain reaction, Western blot analysis, and enzyme-linked immunosorbent assay were conducted to examine the expressions of miR-375, astrocyte elevated gene-1 (AEG-1), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and IL-1ß in Kupffer cells of mice with liver failure as well as after silencing of miR-375. Flow cytometry was used to determine cell apoptosis. During the liver failure process, miR-375, IL-6, TNF-α, and IL-1ß expressions were increased over time, while AEG-1 expression decreased over time in the control, ConA-1h, ConA-3h, and ConA-6h groups. Opposite alternations were observed after silencing of miR-375. Dual-luciferase reporter gene assay showed that AEG-1 was a target gene of miR-375. Flow cytometry determination showed that the ratio of apoptotic Kupffer cells decreased after silencing of miR-375. Overexpression of AEG-1 could rescue the suppression of IL-6, TNF-α, and IL-1ß expressions in Kupffer cells after the short-term induction of ConA and further inhibit cell apoptosis. Our study provides evidence that miR-375 could regulate Kupffer cells to improve immune function in mice with liver failure.

New-onset hyperglycemia immediately after liver transplantation: A national survey from China Liver Transplant Registry.

BACKGROUND: New-onset hyperglycemia (NOH) is a common phenomenon after liver transplantation (LT), but its impact on clinical outcomes has not yet been fully assessed. We aimed to evaluate the etiology and prognosis of NOH within 1 month after LT. METHODS: The data of 3339 adult patients who underwent primary LT from donation after citizen death between January 2010 and June 2016 were extracted from China Liver Transplant Registry database and analyzed. NOH was defined as fasting blood glucose ≥7.0 mmol/L confirmed on at least two occasions within the first post-transplant month with or without hypoglycemic agent. RESULTS: Of 3339 liver recipients, 1416 (42.4%) developed NOH. Recipients with NOH had higher incidence of post-transplant complications such as graft and kidney failure, infection, biliary stricture, cholangitis, and tumor recurrence in a glucose concentration-dependent manner as compared to non-NOH recipients (P < 0.05). The independent risk factors of NOH were donor warm ischemic time >10 min, cold ischemic time >10 h, anhepatic time >60 min, recipient model for end-stage liver disease score >30, moderate ascites and corticosteroid usage (P < 0.05). Liver enzymes (alanine aminotransferase and gamma-glutamyltranspeptidase) on post-transplant day 7 significantly correlated with NOH (P < 0.001). CONCLUSIONS: NOH leads to increased morbidity and mortality in liver recipients. Close surveillance and tight control of blood glucose are desiderated immediately following LT particularly in those with delayed graft function and receiving corticosteroid. Strategic targeting graft ischemic injury may help maintain glucose homeostasis.

Value of Two-Dimensional Shear Wave Elastography for Assessing Acute Liver Congestion in a Bama Mini-Pig Model.

BACKGROUND: To date, liver congestion is one of the most significant clinical diseases. However, few studies have profoundly investigated the development, pathology, and prognosis of the important problems associated with acute hepatic congestion. AIMS: To explore the value of noninvasive two-dimensional shear wave elastography (2D-SWE) for assessing acute liver congestion in an animal model. METHODS: Six healthy Bama mini-pigs were used for this research and randomly divided into the experimental group and control group. We measured the basal liver stiffness (LS) by 2D-SWE and then clamped the inferior vena cava (IVC). LS was measured after 1, 5, 10, and 15 min. We reopened the IVC of experimental group pigs and detected the LS again. All pigs were killed and obtained for a pathological microscopic examination. RESULTS: LS was distinctly increased from 7.03 ± 0.48 to 17.18 ± 3.40 kPa (p < 0.01) within 15 min and reversed to almost normal values of 7.59 ± 0.77 kPa (p < 0.01) within 5 min. In addition, two-dimensional ultrasound images demonstrated the interesting phenomenon of spontaneous echo contrast. Most importantly, the pathologic results of experimental group pigs showed the central veins of the hepatic lobules and hepatic sinusoids were enlarged and filled with numerous erythrocytes; central lobular hepatocytic necrosis and edema were noted. CONCLUSIONS: In conclusion, 2D-SWE is a valuable, reliable, and quantitative approach to successfully assess acute liver congestion, and it is well consistent with histopathological characteristics. Besides, acute liver congestion is an important factor influencing LS that increases LS in a reversible way.

Rupture of hepatic hemangioma with hemoperitoneum due to spontaneous gallbladder perforation: A unique case report.

INTRODUCTION: Hemangiomas are common benign tumors of the liver. Spontaneous rupture is a rare complication, occurring most commonly in giant hemangiomas. Rupture of a hemangioma with hemoperitoneum is a serious development and can be fatal if not managed promptly.The present study reports the unique case of a man who experienced rupture and hemorrhage of a hepatic hemangioma (HH) due to perforation of the gallbladder fundus. After en block resection of the hemangioma and gallbladder using the Pringle maneuver, the patient made an uneventful recovery without complications.To our knowledge, spontaneous rupture of HH secondary to gallbladder perforation has not been reported in the literature. This case highlights a unique, rare cause of ruptured HH and the need to consider appropriate treatment for some hemangiomas to avoid this potentially fatal complication. CONCLUSION: The current case may provide additional support for treatment of HH due to the potential for spontaneous rupture. For patients with ruptured HH, enucleation with the Pringle maneuver is recommended.

Risk factors of choledocholithiasis formation after liver transplantation.

Systematic study of risk factors for biliary stone post-liver transplantation is rarely performed. To investigate the risk factor of choledocholithiasis formation after liver transplantation, we conducted a case-control study. Fourteen patients were selected into a study group. The stones of the bile duct of the patients were confirmed and treated successfully by endoscopic retrograde cholangiopancreatography. For univariate analysis, we selected carefully some potential risk factors such as cold ischemia time, warm ischemia time, and biliary stricture. The results revealed that cold ischemia time and biliary stenosis were significant predictors. But multivariate analysis revealed that only biliary stenosis was a significant risk factor. In conclusion, biliary stenosis is a risk factor of bile duct stones formation after liver transplantation. Endoscopic retrograde cholangiopancreatography is effective and safe in the diagnosis or treatment of bile duct stones after liver transplantation.

Chronic cholecystitis with hilar bile duct stricture mimicking gallbladder carcinoma on positron emission tomography: A case report.

Thickening of the gallbladder wall is observed in patients with gallbladder carcinoma, as well as in those with chronic cholecystitis. It is difficult to distinguish between benign and malignant gallbladder wall thickening with conventional diagnostic imaging techniques, such as abdominal ultrasonography (US), computed tomography (CT) and magnetic resonance imaging (MRI), particularly in patients with bile duct strictures. Currently, the fluorine-18 2-fluorodeoxyglucose positron emission tomography/CT (F-18 FDG PET/CT) scan is widely used in the differentiation of cholecystitis from gallbladder carcinoma. However, the F-18 FDG PET/CT scan may also be responsible for false-positive diagnosis. This case report focuses on a 74-year-old male who presented with thickening of the gallbladder wall and hilar bile duct stricture, originally misdiagnosed as gallbladder carcinoma by US and MRI. F-18 FDG PET/CT also demonstrated increased activity. This case was ultimately proven to be chronic cholecystitis by postoperative pathological examination and it is presented in order to emphasize the significance of considering the possibility of false-positive diagnosis by PET/CT, as a result of inflammatory lesions. Therefore, PET/CT should not be considered the gold standard for the discrimination between benign and malignant gallbladder wall thickening.

Genome-wide analysis of aberrant DNA methylation for identification of potential biomarkers in colorectal cancer patients.

BACKGROUND: Colorectal cancer is one of the leading causes of mortality worldwide. Genome wide analysis studies have identified sequence mutations causing loss-of-function that are associated with disease occurrence and severity. Epigenetic modifications, such DNA methylation, have also been implicated in many cancers but have yet to be examined in the East Asian population of colorectal cancer patients. METHODS: Biopsies of tumors and matched non-cancerous tissue types were obtained and genomic DNA was isolated and subjected to the bisulphite conversion method for comparative DNA methylation analysis on the Illumina Infinium HumanMethylation27 BeadChip. RESULTS: Totals of 258 and 74 genes were found to be hyper- and hypo-methylated as compared to the individual's matched control tissue. Interestingly, three genes that exhibited hypermethylation in their promoter regions, CMTM2, ECRG4, and SH3GL3, were shown to be significantly associated with colorectal cancer in previous studies. Using heatmap cluster analysis, eight hypermethylated and 10 hypomethylated genes were identified as significantly differentially methylated genes in the tumour tissues. CONCLUSIONS: Genome-wide methylation profiling facilitates rapid and simultaneous analysis of cancerous cells which may help to identify methylation markers with high sensitivity and specificity for diagnosis and prognosis. Our results show the promise of the microarray technology in identification of potential methylation biomarkers for colorectal cancers.

Radical resection and outcome for malignant tumors of the pancreatic body and tail.

AIM: To analyze the factors influencing radical (R0) resection rate and surgical outcome for malignant tumor of the pancreatic body and tail. METHODS: The clinical and operative data and follow-up results of 214 pancreatic body and tail cancer patients were analyzed retrospectively. RESULTS: One hundred and twenty/214 pancreatic body and tail cancer patients underwent surgical treatment; the overall resection rate was 59.2% (71/120), and the R0 resection rate was 40.8% (49/120). Compared with non-R0 treatment, the patients receiving an R0 resection had smaller size tumor (P<0.01), cystadenocarcinoma (P<0.01), less lymph node metastasis (P<0.01), less peri-pancreatic organ involvement (P<0.01) and earlier stage disease (P<0.01). The overall 1-, 3- and 5-year survival rates for pancreatic body and tail cancer patients were 12.7% (25/197), 7.6% (15/197) and 2.5% (5/197), respectively, and ductal adenocarcinoma patients had worse survival rates [15.0% (9/60), 6.7% (4/60) and 1.7% (1/60), respectively] than cystadenocarcinoma patients [53.8% (21/39), 28.2% (11/39) and 10.3% (4/39)] (P<0.01). Moreover, the 1-, 3- and 5-year overall survival rates in patients with R0 resection were 55.3% (26/47), 31.9% (15/47) and 10.6% (5/47), respectively, significantly better than those in patients with palliative resection [9.5% (2/21), 0 and 0] and in patients with bypass or laparotomy [1.2% (1/81), 0 and 0] (P<0.01). CONCLUSION: Early diagnosis is crucial for increasing the radical resection rate, and radical resection plays an important role in improving survival for pancreatic body and tail cancer patients.

The value of serum alpha-fetoprotein in predicting tumor recurrence after liver transplantation for hepatocellular carcinoma.

BACKGROUND: Many patients with hepatocellular carcinoma (HCC) who undergo liver transplantation (LT) subsequently develop tumor recurrence; this is the main factor affecting long-term survival after LT. Factors associated with tumor recurrence should be determined to improve the outcome of LT. The purpose of the study was to evaluate the value of alpha-fetoprotein (AFP) in forecasting tumor recurrence after LT for patients with HCC. METHODS: AFP data before and after LT for 97 patients with HCC who underwent LT in our center were analyzed retrospectively. RESULTS: The mean follow-up time was 17.1 +/- 2.1 months for all 97 patients, overall tumor recurrence rate was 32.9% (32/97), and mean recurrence time was 7.2 +/- 3.2 months. The most common tumor recurrence sites were liver, lung, skeleton, and other sites. Pre-transplant AFP levels >400 ng/ml were associated with higher tumor recurrence. Post-transplant AFP levels not decreasing to

[Factors influencing the long-term survival in pancreatic carcinoma patients after radical resection].

OBJECTIVE: To investigate the factors influencing the long-term survival of pancreatic carcinoma patients after radical resection. METHODS: The data of 184 pancreatic carcinoma patients with radical resection were analyzed retrospectively. Analysis of the prognostic factors influencing the long-term survival was performed using Cox proportional hazard regression model. RESULTS: The overall 1-, 3- and 5-year survival rates in this group were 61.7%, 29.0% and 14.3%, respectively. They were 78.0%, 38.4% and 25.7%, respectively, for the patients with a tumor < 3 cm in diameter, significantly better than those with a tumor >or= 3 cm (52.8%, 22.7% and 7.2%, respectively, P < 0.05). Moreover, the 1-, 3- and 5-year survival rates were 67.6%, 30.5% and 17.4%, respectively, in the patients without lymph node involvement, much longer than that in those with lymph node metastasis (37.1%, 20.6% and 0, respectively, P < 0.05). Multivariate analysis by Cox proportional hazard regression model revealed that the tumor size (P < 0.05) and lymph node metastasis (P < 0.01) significantly influenced the long-term survival of the patients. CONCLUSION: Tumor size and lymph node metastasis are significant factors influencing the long-term survival of pancreatic carcinoma patients with radical resection. Therefore, early diagnosis and radical resection are the key points to improve treatment outcome.

Pretreatment of hypertonic saline can increase endogenous interleukin 10 release to attenuate hepatic ischemia reperfusion injury.

Ischemia-reperfusion (I/R) injury of the liver occurs in many clinical cases. Many steps are associated with hepatic I/R injury, including the release of many inflammatory molecules and infiltration of neutrophils into the liver. Recent studies revealed that hypertonic saline (HTS) has a strong anti-inflammatory effect and can inhibit a varity of neutrophil functions. So pretreatment with HTS may attenuate the liver injury associated with I/R. In this study, rats were divided into three groups: the sham group (S group), hepatic I/R group (I/R group), and HTS pretreatment group (HTS group). Serum ALT and myeloperoxidase (MPO) activity were determined. Serum tumor necrosis factor alpha (TNF-alpha) and interleukin 10 (IL-10) were determined by enzyme-linked immunosorbent assay (ELISA). Reverse transcription polymerase chain reaction analysis was used to assess the mRNA expressions of TNF-alpha and IL-10. Protein expressions of TNF-alpha, IL-10, STAT3, and phosphorylated STAT3 were analyzed by Western blot. Results showed that HTS pretreatment can augment the release of endogenous IL-10 by activating STAT3 in the process of hepatic I/R injury. Serum ALT levels, MPO activity in liver, generation of TNF-alpha, and infiltration of neutrophils in liver were inhibited in the HTS group. So we concluded that HTS pretreatment attenuates hepatic I/R injury by increasing the release of endogenous IL-10.

[Prevention of central pontine myelinolysis in rats by early treatment with dexamethasone].

OBJECTIVE: To explore the effect and mechanism of dexamethasone (DEX) in the prevention of central pontine myelinolysis (CPM) in rats. METHODS: Hyponatremia was induced in rat by subcutaneous injection of Vasopressin Tannate and intraperitoneal injection of 2.5% dextrose in water for 3 d, the rats of Group A received a bolus of 1 mol/L NaCl (2 ml/kg) and DEX (5 mg/kg) simultaneously at the 4th day; the rats of Group B were treated with DEX after 24 h of the injection of 1 mol/L NaCl; the rats in Group C received a bolus of 1 mol/L NaCl and saline simultaneously; Group D was the control group. The demyelinative lesions were evaluated by myelin staining. The Evans blue (EB) contents of brain were detected to evaluate the blood-brain-barrier permeability after rapid correction of hyponatremia. The expression of inducible nitric oxide synthase (iNOS) in brains was evaluated by Western blotting. RESULT: CPM was induced successfully in rats. The EB contents of Group A, B and C had no significant difference at 0 h after injection of hypertonic saline compared with Group D. The EB contents of Group C began to increase significantly at 6 h after injection of hypertonic saline, peaked at 24 h; the expression of iNOS in brains began to increase after 3 h after the rapid correction of hyponatremia. The rate of morbidity in Group C was 66.7%. The demyelinative lesions were rarely seen in Group A, the EB contents of brain decreased significantly compared with Group C at the same time point (P<0.05), the iNOS expression was also inhibited. DEX could not prevent the attack of CPM at Group B, the rate of morbidity (75%) had no significant difference compared with Group C (P>0.05). CONCLUSION: Early treatment with DEX can protect blood-brain-barrier and inhibit the expression of iNOS to prevent the attack of CPM.

[Effects of hypertonic saline on expression of heme oxygenase enzyme-1 in hepatic ischemia/reperfusion injury rats].

OBJECTIVE: To explore the effect of the pretreatment with hypertonic saline (HTS) in the hepatic ischemia/reperfusion (I/R) injury. METHODS: Twenty-five SD rats were randomly divided into sham operation group, heme oxygenase-1 (HO-1) blocker ZnPP group, I/R group, HTS pretreatment group and ZnPP intervention group (n=5). The rat model of partial hepatic I/R injury was reproduced by isolating the portal venous and hepatic arterial branches to the left and median hepatic lobes, and they were occluded with a microvascular clamp for 30 minutes, followed by reperfusion. In HTS pretreatment group, the rats received 4 ml/kg volume of HTS (7.5%) intravenous 1 hour before the occlusion of the vessels. The rats were sacrificed 6 hours after reperfusion. The levels of alanine aminotransferase (ALT) and tumor necrosis factor-alpha (TNF-alpha) in serum, liver myeloperoxidase (MPO) activity and liver endothelin-1 (ET-1) were determined. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to examine the expression of HO-1 in the liver. The pathological changes in the liver, including hepatic sinusoid, were evaluated in hematoxylin and eosin (HE)-stained sections and transmission electron microscopy (TEM) of liver specimens. The effect of HTS pretreatment was also assessed in the rats pretreated with ZnPP. RESULTS: The levels of serum ALT and TNF-alpha, ET-1 and MPO activity in hepatic tissues increased after hepatic I/R injury (all P<0.01), and expression of HO-1 mRNA and protein were also increased. RT-PCR and Western-blot revealed that the expression of HO-1 in the liver was upgraded significantly, and the ALT level, serum TNF-alpha, liver MPO activity and liver ET-1 were suppressed significantly after I/R injury in the HTS pretreatment group. In this group, there were moderate swelling of hepatocytes and mild neutrophils infiltration in the liver. The hepatic microcirculatory dysfunction was ameliorated. All these findings showed that HTS pretreatment produced the effect of prevention on hepatic I/R injury. However, the adjunctive infusion of ZnPP abrogated the beneficial effects. CONCLUSION: Pretreatment of HTS has the effect of the prevention of hepatic I/R injury by promotion of the expression of HO-1.

[The possible causes of central pontine myelinolysis after liver transplantation].

OBJECTIVE: To sum up the clinical characteristics of patients with central pontine myelinolysis (CPM) after orthotopic liver transplantation (OLT) and to document the possible causes on CPM. METHODS: 142 patients' data with OLT between January 1999 to May 2003 were analyzed retrospectively. The following risk factors during preoperation were analyzed between patients with and without CPM: primary liver disease, preoperative serum sodium level, magnesium level and plasma osmolality, fluctuation degree of serum sodium concentration, and immunosuppressive drugs level etc. RESULTS: A total of 13 (9.2%) neurologic symptoms appeared in 142 patients post operation, including 5 cases (3.5%) with CPM and 8 cases (5.6%) with cerebral hemorrhage or infarct. 2 patients who developed CPM after OLT had hyponatremia history before operation (serum sodium <130 mmol/L), and the mean serum sodium level was (130.6 +/- 5.54) mmol/L. The serum sodium level was significantly lower in CPM than that of patients without neurologic complication or with cerebral hemorrhage/infarct (P <0.05). The rises of serum sodium perioperative 48 h after OLT in patients with CPM was significantly greater than that in patients with cerebral hemorrhage/infarct or no neurologic complication (19.5 +/- 6.54) mmol/L, (10.1 +/- 6.43) mmol/L, (4.5 +/- 4.34) mmol/L, respectively, (P < 0.05). Plasma osmolality increased greatly postoperatively in patients with CPM. Hypomagnesemia was noted in all patients perioperatively, but there was not significant difference among groups. The duration of operation in CPM was longer than in others (492 +/- 190.05) min (P <0.05). Cyclosporin A (CsA) levels were normal in all patients, but there was significant difference between patients with and without neurologic complication (P <0.05). CONCLUSIONS: CPM may be more prevalent following liver transplantation. Although the diagnosis of CPM after OLT can be made by complete neurologic evaluation including magnetic resonance imaging (MRI) of the head, the mortality is still very high. The occurrence of CPM may be associated with hyponatremia, rapid rise of serum sodium concentration, postoperative increase of plasma osmolality, the duration of operation and high CsA levels.

Possible causes of central pontine myelinolysis after liver transplantation.

AIM: To sum up the clinical characteristics of patients with central pontine myelinolysis (CPM) after orthotopic liver transplantation (OLT) and to document the possible causes of CPM. METHODS: Data of 142 patients undergoing OLT between January 1999 to May 2003 were analyzed retrospectively. Following risk factors during perioperation were analyzed in patients with and without CPM: primary liver disease, preoperative serum sodium level, magnesium level and plasma osmolality, fluctuation degree of serum sodium concentration, and immunosuppressive drug level, etc. RESULTS: A total of 13 (9.2%) neurologic symptoms appeared in 142 patients post-operation including 5 cases (3.5%) with CPM and 8 cases (5.6%) with cerebral hemorrhage or infarct. Two patients developing CPM after OLT had a hyponatremia history before operation (serum sodium<130 mmol/L), their mean serum sodium level was 130.6 +/- 5.54 mmol/L. The serum sodium level was significantly lower in CPM patients than in patients without neurologic complications or with cerebral hemorrhage/infarct (P<0.05). The increase in serum sodiumduring perioperative 48 h after OLT in patients with CPM was significantly greater than that in patients with cerebral hemorrhage/infarct but without neurologic complications (19.5 +/- 6.54 mmol/L, 10.1 +/- 6.43 mmol/L, 4.5 +/- 4.34 mmol/L, respectively, P<0.05). Plasma osmolality was greatly increased postoperation in patients with CPM. Hypomagnesemia was noted in all patients perioperation, but there were no significant differences between groups. The duration of operation on patients with CPM was longer than that on others (492 +/- 190.05 min, P<0.05). Cyclosporin A (CsA) levels were normal in all patients, but there were significant differences between patients with or without neurologic complications (P<0.05). CONCLUSION: CPM may be more prevalent following liver transplantation. Although the diagnosis of CPM after OLT can be made by overall neurologic evaluations including magnetic resonance imaging (MRI) of the head, the mortality is still very high. The occurance of CPM may be associated with such factors as hyponatremia, rapid rise of serum sodium concentration, plasma osmolality increase postoperation, the duration of operation, and high CsA levels.