Hepatocellular carcinoma (HCC) is one of the most aggressive types of cancer. Although it has a high mortality rate, there is currently no effective treatment for HCC. Lenvatinib has traditionally been used as the first-line treatment for advanced HCC (aHCC); however, resistance to this therapy is common. It can be difficult to select effective second-line drugs to overcome lenvatinib resistance when treating aHCC. For patients with aHCC, poor treatment efficacy can result in patients missing the optimal treatment window and can lead to an irreversible situation. Lenalidomide has begun to be used to treat HCC; however, to the best of our knowledge, its efficacy in patients with lenvatinib-resistant HCC remains to be reported on in the literature. The present case report, to the best of our knowledge, describes the first case in the literature of a patient with lenvatinib-resistant aHCC who achieved a partial response after the treatment regimen was switched to lenalidomide. The present case report provides a promising novel route for the treatment of lenvatinib-resistant HCC.

In the article cited above, the nomenclature for a novel long noncoding RNA (lncRHL, regulator of hyperlipidemia) was similar to that used for a novel long noncoding RNA in the 2021 Cell Proliferation article below (lnc-RHL, regulator of hepatic lineages).

Objective:To investigate the effects of long non-coding RNA Hox transcript antisense RNA (lncRNA HOTAIR) on proliferation and migration ability of pancreatic cancer cells by targeting microRNA-520g-3p (microRNA-520g, miR-520g-3p).Methods:Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-520g-3p and HOTAIR in pancreatic cancer cells. HOTAIR small interfering RNA was transfected into SW1990 pancreatic cancer cells. The normal control (NC) group, siHOTAIR-1 group and siHOTAIR-2 group were set up. Cell counting kit-8 (CCK-8) assay was used to detect cell proliferation ability. The scratch healing experiment was used to detect cell migration ability. The dual luciferase reporter gene experiment was used to verify the relationship between miR-520g-3p and HOTAIR targeting regulation. The quantitative data between the two groups were compared by t test. Results:qRT-PCR showed that the level of HOTAIR mRNA in SW1990 was significantly higher than that in human normal pancreatic ductal epithelial cells human pancreatic duct epithelial (HPDE) (3.08±0.33 vs. 1.57±0.17, t=29.830, P<0.01). The expression of miR-520g-3p in SW1990 was significantly lower than that in HPDE (0.93±0.11 vs. 2.63±0.24, t=22.410, P<0.01). HOTAIR small interfering RNA was transfected into SW1990, and at72 h, A value in the NC group was significantly higher than that in siHOTAIR-1 group and siHOTAIR-2 group (3.52±0.99 vs. 1.92±0.23, 2.36±0.58, t=6.340, 5.150, P<0.01). At 48 h after down-regulating the HOTAIR expression, the percentage of wound healing area in the NC group was significantly higher than that in the siHOTAIR-1 group and the siHOTAIR-2 group [(85.37±9.82)% vs. (50.56±5.82)%, (35.28±6.86)%, t=7.760, 14.750, P<0.01]. The dual luciferase reporter gene indicated that miR-520g-3p could significantly reduce the luciferase activity of wild-type HOTAIR vector. After down-regulating the expression level of miR-520g-3p, the proliferation and migration ability of SW1990 cells was significantly increased as compared with that in the control group ( t=13.190, 12.480, P<0.01). Conclusion:HOTAIR can influence the proliferation and migration ability of pancreatic cancer cells by targeting and negatively regulating miR-520g-3p.

Intrahepatic cholangiocarcinoma (ICC) is a highly lethal hepatobiliary neoplasm, which originates from the bile ducts proximal to the second-order division. ICC can be anatomically divided into two subtypes: the large duct type (mucin-production ICC, muc-ICC) and the small duct type (mixed-ICC) origins from hepatic progenitor cells (HPCs). The immunoreactivity of S100P and neural cell adhesion molecule (NCAM) are useful biomarkers to distinguish the two subtypes. In this study, we report a difficult-to-diagnose case of metastatic retroperitoneal tumor of occult hepatolithiasis-associated ICC. Besides, this case was both positive for S100P and NCAM, considered as a rare muc-ICC with the HPCs features. Tumor whole exome sequencing detection results by Genetron (China) revealed that there were 41 gene mutations in this patient. The SMAD4-p.His530ThrfsTer47 and KRAS-p.Gly12Val mutation might promote the occurrence and distant metastasis of the tumor.

Dysregulation of hepatic VLDL secretion contributes to the pathogenesis of metabolic diseases, such as nonalcoholic fatty liver disease (NAFLD) and hyperlipidemia. Accumulating evidence has suggested that long noncoding RNAs (lncRNAs) had malfunctioning roles in the pathogenesis of NAFLD. However, the function of lncRNAs in controlling hepatic VLDL secretion remains largely unillustrated. Here, we identified a novel lncRNA, lncRNA regulator of hyperlipidemia (lncRHL), which was liver-enriched, downregulated on high-fat diet feeding, and inhibited by oleic acid treatment in primary hepatocytes. With genetic manipulation in mice and primary hepatocytes, depletion of lncRHL induces hepatic VLDL secretion accompanied by decreased hepatic lipid contents. Conversely, lncRHL restoration reduces VLDL secretion with increased lipid deposition in hepatocytes. Mechanistic analyses indicate that lncRHL binds directly to heterogeneous nuclear ribonuclear protein U (hnRNPU), and thereby enhances its stability, and that hnRNPU can transcriptional activate Bmal1, leading to inhibition of VLDL secretion in hepatocytes. lncRHL deficiency accelerates the protein degradation of hnRNPU and suppresses the transcription of Bmal1, which in turn activates VLDL secretion in hepatocytes. With results taken together, we conclude that lncRHL is a novel suppressor of hepatic VLDL secretion. Activating the lncRHL/hnRNPU/BMAL1/MTTP axis represents a potential strategy for the maintenance of intrahepatic and plasma lipid homeostasis.

Objective:To detect the expression of receptor for activated C kinase 1 (RACK1) in cholangiocarcinoma cells, and to explore its effect on the biological phenotype of cholangiocarcinoma cells.Methods:Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) and Western blotting were used to detect the expression level of RACK1 in human normal bile duct epithelial cells HIBEC, 2 cholangiocarcinoma cell lines RBE and HUH28. The t test was used to analyze the difference in the RACK1 expression. Lentivirus was used to construct a stable low-expression strain of RACK1, which was divided into sh-RACK1-1, sh-RACK1-2 and negative control (NC-RACK1) group. The transfection efficiency was tested by qPCR. The invasion and migration abilities of cholangiocarcinoma cells were tested by Transwell experiment. T test was used to analyze the migration and invasion ability of cholangiocarcinoma cells after knocking down RACK1. Results:The qPCR results showed that the expression level of RACK1 in RBE and HUH28 cholangiocarcinoma cell lines (3.45±0.92, 2.96±0.33) was significantly higher than that in HIBEC cells (1.73±0.58, t=21.965, 24.153, P<0.01). Western blotting showed that the expression level of RACK1 in RBE and HUH28 cells was significantly higher than that in HIBEC cells. The results of Transwell migration experiment suggested that the number of cells penetrating the membrane in the sh-RACK1-1 and RBE RACK1-2 groups in RBE cells was [(261.89±8.65), (231.07±9.73)/field of view], significantly less than that in the NC-RACK1 group [(430.19±10.15)/field of view, t=30.684, 24.380, P<0.01]. The number of cells penetrating the membrane in the sh-RACK1-1 and sh-RACK1-2 groups in HUH28 cells [(253.57±7.28)/field of view, (210.11±6.09)/field of view] was significantly less than in the NC-RACK1 group [(424.78±8.03)/field of view] ( t=35.442, 36.781, P<0.01). The results of the invasion experiment showed that the number of cells in the sh-RACK1-1 and sh-RACK1-2 groups in RBE cells was [(52.15±1.43) and (60.58±2.01)/field of view] was significantly lower than that in NC-RACK1 group [(125.39±4.03)/field of view, t=9.935, 12.566, P<0.05]. The number of cells passing through the membrane in the sh-RACK1-1 and sh-RACK1-2 groups [(60.91±1.30) and (51.19±0.96)/field of view] in HUH28 cells was less than in the NC-RACK1 group [(150.88±7.28)/field of view] ( t=13.554, 15.328, P<0.01). Conclusion:RACK1 is up-regulated in cholangiocarcinoma cells. The down-regulation of the RACK1 expression can inhibit the migration and invasion of cholangiocarcinoma cells. RACK1 can promote the occurrence and development of cholangiocarcinoma.

Objective:To detect the expression of GINS complex subunit 4 (GINS4) protein in hepatocellular carcinoma tissues and observe its effect on the invasion and proliferation of hepatocellular carcinoma cells.Methods:From September 2019 to December 2020 in our hospital, 60 patients with hepatocellular carcinoma were collected during surgery and hepatocellular carcinoma tissues and corresponding para-cancerous tissue specimens were selected as the observation group and the control group. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blotting was used to detect the expression of GINS4 in hepatocellular carcinoma tissues, use specific short hairpin RNA (shRNA) to construct low-expressing stable hepatocellular carcinoma cell lines, and divide them into experimental groups (shRNA group) and the control group (NC group); Transwell tested its ability to invade HepG2 cells; colony formation test and cell counting kit (CCK-8) test tested its proliferation ability on HepG2 cells. Comparison between groups is by paired sample t test. Results:RT-qPCR showed that the expression of GINS4 in hepatocellular carcinoma tissues (2.89±0.11) was higher than that in adjacent tissues (1.34±0.15), and the difference was statistically significant ( t=8.56, P<0.01). Western blot showed that the expression of GINS4 in hepatocellular carcinoma tissue was (1.54±0.76) times higher than that in adjacent tissues, and the difference was statistically significant ( t=2.67, P<0.05). After shRNA transfection of HepG2 cells, GINS4 in the experimental group The expression level was (1.88±0.34) times lower than that of the control group, and the difference was statistically significant ( t=7.43, P<0.05); the Transwell experiment showed that the number of perforated cells in the experimental group was (56.33±3.51) compared with the control group (90.42±1.53) cells were significantly reduced, and the difference was statistically significant ( t=3.34, P<0.01); the colony formation experiment showed that the number of cell clusters in the experimental group (83.33±7.37) was significantly less than that in the control group (345.00±16.64). The difference was statistically significant ( t=15.28, P<0.01); CCK-8 experiment showed that the 3 d absorbance of the experimental group was (1.43±0.11) lower than that of the control group (2.15±0.15), and the difference was statistically significant ( t=6.52, P<0.001). Conclusion:The expression of GINS4 in hepatocellular carcinoma tissues is higher than that in adjacent tissues. Reducing the expression of GINS4 can significantly inhibit the invasion and proliferation of hepatocellular carcinoma cells.

Objective:To investigate the application of nano-fluorescent probes in hepatocellular carcinoma (HCC) blood supply molecular imaging in rats and pave the foundation of double blood supply treatment in HCC.Methods:The HCC models of 12 rats were established by diethylnitrosamine (DEN) chemical induction. After 15 weeks, the magnetic resonance imaging (MRI) and pathological findings were observed. Different nano-fluorescent probes were applied through hepatic artery and portal vein to observe the blood supply molecular images of HCC tissues.Results:A total of 4 rats died at 13-14 weeks, with a survival rate of 66.67% at 15 weeks and the tumor formation rate of 100%. The pathological sections were consistent with the signs of HCC. The arterial 1, 1′-dioctadecy1-3, 3, 3′, 3′-tetramethylindocarbocyanine perchlorate (DiI) fluorescence intensity in tumor of molecular fluorescence imaging [(2.99±0.41)×10 7] was significantly greater than the surrounding liver tissue [(1.06±0.22)×10 7, t=-9.410, P<0.05]; and fluorescence intensity of Alexa fluor 647 in tumor of portal vein [(1.30±0.29)×10 7] was significantly weaker than that of peripheral liver tissue [(2.76±0.38)×10 7, t=7.480, P<0.05]. The slide scan showed that arterial blood supply in tumor tissue was higher than the surrounding liver tissue, and the portal vein blood supply was found in the parenchyma and surrounding areas of the liver tumor. Conclusion:The DEN induction is a mature and stable method to establish the HCC model, and the nano-fluorescent probe can observe the double blood supply for HCC in the microscopic level.

Background: Methylprednisolone (MP) is a synthetic corticosteroid with potent anti-inflammatory and antioxidant properties used as therapy for a variety of diseases. The underlying mechanism of MP to reduce acute pancreatitis still needs to be elucidated.Methods: Twenty-four male C57BL/6 mice (6-8 weeks) were used to establish SAP mouse model by administering an intraperitoneal injection of Cae and LPS. Amylase expression levels of serum and PLF were measured with an amylase assay kit. The concentrations of IL-1β and TNF-α in the serum and PLF were detected by ELISA. The level of pancreatic and lung tissue damage and inflammation was assessed by H&E staining and immunofluorescence staining. Western blot and qPCR were used to detect the expression levels of NLRP3, IL-1β and TNF-αin vivo and in vitro.Results: In this study, we found MP, used in the early phase of SAP, decreased the levels of IL-1β and TNF-α in serum and peritoneal lavage fluids (PLF), reduced the level of serum amylase and the expression of MPO in lung tissue, attenuated the pathological injury of the pancreas and lungs in a dose-dependent manner. The expression of NLRP3 and IL-1β in pancreas and lungs was down-regulated significantly depending on the MP concentration. In vitro, MP reduced the levels of IL-1β and TNF-α by down-regulating the expression of NLRP3, IL-1β and p-NF-κB in isolated peritoneal macrophages. Conclusion: MP can attenuate the injury of pancreas and lungs, and the inflammatory response in SAP mice by down-regulating the activation of NF-κB and the NLRP3 inflammasome.

Various surgical methods impact the prognosis of patients with hepatocellular carcinoma (HCC) differently. However, clinical guidelines remain inconsistent and the relative importance of predictors of survival outcomes requires further evaluation. The present study aimed to rank the importance of predictive factors that impact the survival outcomes of patients with HCC and to compare the prognosis associated with different surgical methods based on data obtained from the Surveillance, Epidemiology and End Results database. To achieve these aims, the present study used a random forest (RF) model to detect important predictive factors associated with survival outcomes in patients with HCC. Cox regression analysis was used to compare different surgery methods. The variables included in the Cox regression model were selected based on the Gini index calculated by the RF model. Using the RF model, the present study demonstrated that surgery method, tumor size and age were the first, second and third most important factors associated with HCC prognosis, respectively. Overall, patients who underwent local tumor destruction [(hazard ratio (HR)=0.48; 95% confidence interval (CI), 0.45-0.51; P<0.001)], wedge or segmental resection (HR, 0.31; 95% CI, 0.29-0.33; P<0.001), lobectomy (HR, 0.29, 95% CI, 0.27-0.31; P<0.001) or liver transplantation (HR, 0.16; 95% CI, 0.14-0.17; P<0.001) demonstrated improved overall survival time compared with those treated with surgery, with a gradual decreasing trend observed in HRs. The present study demonstrated that the surgical method used is the most important predictor of the survival outcomes of patients with HCC. Liver transplantation resulted in the best prognosis for patients with HCC, except for those with undifferentiated tumors or distant metastasis.

Objective To compare the safety and efficacy of laparoscopic technique and open surgery in the treatment of HCC in the right lobe of the liver.Methods A retrospective analysis of data from 163 patients with HCC in the right lobe of the Henan Provincial People's Hospital (Henan Provincial People's Hospital) from January 2017 to January 2019, in which the tumor site was located in the upper right lobe of the liver (group A) 58 For example, 105 patients were located in other parts of the right lobe of the liver (group B). Group A underwent laparoscopic hepatectomy in 23 cases, open abdominal hepatectomy in 35 cases; group B underwent laparoscopic hepatectomy in 45 cases and open abdominal hepatectomy in 60 cases. Groups A and B were compared intra-group, and the laparoscopic group in groups A and B were compared between groups to determine the safety and effectiveness of perioperative laparoscopic resection of right lobe tumor.Results In group A, the difference in tumor diameter, intraoperative blood loss, postoperative gastrointestinal tract time, first day postoperative drainage, postoperative extubation time, and postoperative hospital stay in the LH group compared with the OH group Statistically significant (P<0.05); operative time, hepatic occlusion time, negative rate of tumor margin, postoperative ALT/TBIL and postoperative complications were not statistically significant (P>0.05). In group B, there was a statistically significant difference in gastrointestinal tract time, first-day drainage, postoperative extubation time, and postoperative hospital stay between the LH group and the OH group (P<0.05); The operation time, hepatic occlusion time, intraoperative blood loss, tumor margin negative rate, postoperative ALT/TBIL and postoperative complications were not statistically significant (P>0.05). The LH between A and B was compared. The tumor diameter and operation time were statistically significant (P<0.05). There was no significant difference between the two groups (P>0.05).Conclusion The safety and efficacy of laparoscopic technique in the treatment of HCC in the right lobe of the liver is worthwhile. However, at this stage, compared with other parts of the right lobe of the liver, laparoscopic resection of the upper right lobe of the liver is still a difficult point in the resection of laparoscopic liver cancer. Laparoscopic resection is performed on the selective HCC in the upper right lobe of the liver. Perioperative safety is safe. Sexuality and efficacy are also positive.

Objective To determine whether G protein-coupled receptor 30 (GPR30) agonist G-1 can inhibit the activation of nucleotide oligomerization domain-like receptor family,pyrin domain containing 3 (NLRP3) inflammasomes and the maturation of interleukin-1β (IL-1β).Methods The peritoneal macrophages were extracted from C57BL/6,and then pretreated with 10,30,50 μmol/L G-1.After induction with 50 ng/ml lipopolysaccharide (LPS),macrophages were collected.Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was used to detect the mRNA expression of NLRP3,apoptosis-related microparticle protein (ASC),and IL-1β,and the expression levels of NLRP3,ASC and pro-cysteinyl aspartate-specific protease (Caspase)-1 proteins were detected by Western blotting.The macrophages were pretreated with 1,5,15,30,50 μmol/L G-1,and cells were induced with 50 ng/ml LPS and 5 mmol/L adenosine triphosphate (ATP).The levels of IL-1β and TNF-α in supernatants were determined by enzyme linked immunosorbent assay (ELISA).Results There was no significant difference in the expression of NLRP3,ASC and IL-1β mRNA in macrophages between LPS group and G-1 plus LPS group.There was no significant difference in the expression of NLRP3,ASC and pro-Caspase-1 proteins between LPS group and G-1 plus LPS group.The ELISA results showed that with the increase of G-1 concentration,the content of IL-1β (3 062.42 ± 12.10,1 709.29 ± 25.31,653.27 ± 35.66,144.23 ± 4.18,98.40 ± 2.02) decreased gradually in G-1 plus LPS group as compared with the LPS + ATP group (2 714.73 ± 107.05),and the difference was statisti cally significant when G-1 concentration ≥5 μmol/L (P ＜0.05),while the level of TNF-α in G-1 plus LPS group [(361.93 ± 10.66),(394.62 ± 20.46),(444.17 ± 14.80),(444.73 ± 19.21) and (366.41 ±27.04)] showed no significant difference from the LPS + ATP group (412.38 ± 1.59) (P ＞ 0.05).Conclusion GPR30 agonist G-1 can specifically inhibit the activation of NLRP3 inflammasome and the maturation of IL-1β.

Objective To compare the clinical effects of laparoscopic liver partial resection and open hepatectomy at different stages on liver disease during perioperative period and evaluate its safety . Methods Sevnty-seven patients with partial hepatectomy admitted to People 's Hospital of Zhengzhou University from 2014 to 2017 were retrospectively analyzed .According to the operation method , the patients were divided into laparoscopic group and open surgery group .In laparoscopic group , the study period was from January 2014 to December 2015, and the proficiency period was from January 2016 to December 2017.The operation time, intraoperative bleeding volume, leison diameter, liver function, fasting time, hospital stay and complications were compared .Results In learning laparoscopic group , most of the resected lesions were located in superficial part of the hepatic marginal segment (Ⅱ、Ⅲ、Ⅴ、Ⅵsegment);and in skilled laparoscopic group , the resected lesions could be located in all parts of the liver.The length of hospitalization in learning laparoscopic group [(6.6 ±2.5) d] was shorter than that in open surgery group [ (10.7 ±2.3) d] , and the difference was significant ( P<0.01) .There were no significant differences in operation time , intraoperative bleeding volume , lesion diameter , liver function index , fasting time or complications between open surgery group and learning laparoscopic group ( P>0.05 ) .The operation time of skilled laparoscopic group [ ( 104.6 ±32.4 ) min ] was shorter than that of open surgery group [ ( 140.9 ±32.5 ) min ] , the intraoperative bleeding volume [ ( 139.3 ± 43.5) ml] was less than that of open surgery group [(175.6 ±56.3) ml], the fasting time [(1.5 ± 0.6) d] was shorter than that of open surgery group [(2.0 ±0.5) d], and the hospitalization time [(6.5 ±2.1) d] was shorter than that of open surgery group [(10.8 ±1.8) d], and the differences were significant ( P<0.05) .There was no significant difference in the diameter of lesions , liver function or complications after surgery between skilled laparoscopy group and open surgery group ( P>0.05 ) . Conclusions After the curve learning period , the indications of laparoscopic surgery in People 's Hospital of Zhengzhou University are gradually expanded .The operation time of laparoscopic partial hepatectomy is significantly shortened , the amount of bleeding during the operation is reduced , and the recovery is fast .The safety and effectiveness of laparoscopic partial hepatectomy is gradually prominent .

目的 探讨白蛋白-胆红素(albumin-bilirubin,ALBI)评分与吲哚菁绿(indocyanine green,ICG)试验在预测巴塞罗那肝癌分期系统(Barcelona Clinic Liver Cancer,BCLC)分期B期肝癌患者肝切除术后肝功能衰竭(posthepatectomy liver failure,PHLF)中的价值.方法 179例BCLC分期B期肝癌患者,术后5d发生PHLF 49例为PHLF组,未发生PHLF 130例为对照组.记录2组大范围肝切除术比率及术前ALBI评分、ICG试验15 min代谢率(ICG test at 15 min,ICG-R 15)、凝血酶原时间(prothrombin time,PT)等指标,多因素logistic回归分析BCLC分期B期肝癌患者发生PHLF的危险因素,绘制ROC曲线评估ALBI评分、ICG-R15预测PHLF发生的效能.结果 观察组大范围肝切除术比率(53.17％)、术前ALBI评分[(-2.31±0.75)分]、ICG-R15[(9.20(5.45,14.55)％]较对照组[16.15％、(-2.83±0.63)分、5.90(3.70,8.48)％]高(P＜0.05),PT[(16.18±3.36)s]较对照组[(13.53±1.56)s]延长(P＜0.05);多因素logistic回归分析结果显示,PT延长(OR=1.690,95％CI:1.355～2.108,P＜0.001)、大范围肝切除术(OR=6.946,95％CI:2.816～17.130,P＜0.001)、ALBI＞-1.39分(OR=3.009,95％CI:1.322～7.264,P=0.009)是BCLC分期B期肝癌患者发生PHLF的危险因素;ROC曲线显示,ALBI评分(最佳截断值为-2.387分)联合PT(最佳截断值为15.550 s)预测小范围肝切除术患者发生PHLF的AUC[0.839(95％CI:0.742～0.936,P＜0.001)]高于ICG-R15(最佳截断值为7.950％)联合PT[0.771(95％CI:0.642～0.900,P＜0.001)](P＜0.05).结论 术前ALBI评分＞-1.39分是BCLC分期B期肝癌患者发生PHLF的危险因素,ALBI评分联合PT预测小范围肝切除术BCLC分期B期肝癌患者发生PHLF的效能优于ICG-R15联合PT.

Objective To explore the expression and clinical significance of long non-coding RNA (lncR)-MEG3 in patients with acute pancreatitis.Methods In 63 patients with acute pancreatitis,21 patients with severe acute pancreatitis (SAP group) and 42 patients with mild acute pancreatitis (MAP group) were detected the levels of serum lncR-MEG3,C-reactive protein (CRP) and D-Dimer by real-time RT-PCR.The diagnostic efficiacies of single or joint detection of lncR-MEG3,CRP and D-Dimer were analyzed by ROC.Results The level of lncR-MEG3 was significantly lower and the levels of CRP and D-Dimer were significantly higher in SAP group (3.46 (2.53,5.64),119.7(113.3,129.3) mg/L,2.95(2.39,3.75) mg/L) than those in MAP group (7.12 (6.54,7.93),63.1(39.2,70.3) mg/L,1.17 (0.84,1.53) mg/L) (P＜0.05).When the optimal cut-off values of lncR-MEG3,CRP and D-Dimer were 6.34(0.86,0.67),74.2(0.86,0.81) and 2.34(0.81,0.86),the AUCs for the diagnosis of SAP were 0.797(95％CI:0.673-0.921,P＜0.05),0.814 (95％CI:0.707-0.921,P＜0.05) and 0.846 (95％CI:0.734-0.958,P＜0.05),the sensitivities were 86％,86％ and 81％,and the specificities were 67％,81％ and 86％,respectively.The AUC of joint detection of lncR-MEG3,CRP and D-Dimer for the diagnosis of SAP was 0.908 (95％CI:0.838-0.979,P＜0.05),the sensitivity was 91％ and the specificity was 86％.The AUC of joint detection was significantly higher than that of single detection of lncR-MEG3,CRP and D-Dimer (P ＜ 0.05).There were no significant differences in the sensitivity or specificity between the joint detection and single detection (P＞ 0.05).Conclusion Serum lncR-MEG3,CRP and D-Dimer levels can reflect the severity of acute pancreatitis,lncR-MEG3 is probably a potential biochemical marker for the differential diagnosis of MAP and SAP.

Objective To study the effects of puerarin on the proliferation and apoptosis of human cholangiocarcinoma (RBE) cells and the expression of related genes.Methods RBE cells cultured were treated with puerarin at different concentrations (0,100,250,500,1 000,1 500 μg/ml).And then methyl thiazol tetrazolium (MTr) assay was used to measure cell viability.Flow cytometry (FCM) was used to detect the apoptosis of RBE cells treated with puerarin at (500,1 000 μg/ml) concentrations.The expression levels of B cell lymphoma/leukemia-2 (bcl-2) and bcl-2 associated X protein (bax) genes were detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR).Western blotting was used to detect the expression of apoptosis-related proteins.Results MTT assay revealed that puerarin (500,1 000,1 500 pg/ml) inhibited the proliferation of RBE cells in a time-and dose-dependent manner,with inhibition rate at 12,24 and 48 h being (8.14 ± 1.85) ％,(12.57 ± 3.10)％ and (18.26 ±2.74)％ (P=0.000,0.005,0.003),(22.38±2.77)％,(29.14 ±3.85)％and (37.43±2.11)％ (P=0.005,0.007,0.002),and (35.47 ±3.75)％,(40.81 ±2.52)％ and (52.29 ± 3.98) ％ (P =0.007,0.000,0.002) respectively.After RBE cells were treated with puerarin at (500,1 000 μg/ml) for 24 h,the apoptosis rate [(14.21 ± 2.53)％ and (22.87 ± 2.74)％] was increased greatly as compared with the control group [(3.246 ± 2.25) ％] (P =0.013,0.008).The expression of bax was up-regulated (2.37 ± 0.17 and 3.78 ± 0.41) times as compared with the control group (0.98 ± 0.04) (P =0.005,0.003),and the expression of bcl-2 was down-regulated (1.57 ± 0.14 and 2.86 ±0.55) times as compared with the control group (0.99 ±0.03) (P =0.002 and 0.008).The expression levels of Cleaved-Caspase-8,Cleaved-Caspase-9 and bax protein were increased (P =0.015,0.003 and 0.008) and those of bcl-2 protein were decreased (P =0.021).Conclusion Puerarin can inhibit the proliferation of RBE cells and promote the apoptosis.

The rapid production and release of a large number of inflammatory cytokines can cause excessive local and systemic inflammation in severe acute pancreatitis (SAP) andmultiple organ dysfunction syndrome (MODS), especially pancreatitis-associated acute lung injury (P-ALI), which is themain causeof early death in patientswith SAP.TheNLRP3 inflammasome plays an important role in the maturation of IL-1β and the inflammatory cascade. Here, we established a model of SAP using wild-type (NLRP3) and NLRP3 knockout (NLRP3) mice by intraperitoneal injections of caerulein (Cae) and lipopolysaccharide (LPS). Pathological injury to the pancreas and lungs, the inflammatory response, and neutrophil infiltration were significantly mitigated in NLRP3 mice. Furthermore, INF-39, anNLRP3 inflammasome inhibitor, could reduce the severity of SAP and P-ALI in a dose-dependentmanner. Our results suggested that SAP and P-ALI were alleviated by NLRP3 deficiency in mice, and thus, reducing NLRP3 expression may mitigate SAP-associated inflammation and P-ALI.

AIM To elucidate the underlying mechanism that microRNA-22 (miR-22) promotes the apoptosis of rat pancreatic acinar cells (AR42J) and the elements that regulate the expression of miR-22. METHODS One hundred nanomoles per liter of caerulein (Cae) was administrated to induce the apoptosis of AR42J cells and the apoptosis rate was detected by flow cytometry analysis. An amylase assay kit was used to measure the amylase expression level in the supernatant. Quantitative real-time PCR (qRT-PCR) was adopted to measure miR-22 expression. We used online tools to predict the potential transcription promoter of miR-22 and the binding sites, which was further identified by using luciferase reporter analysis, chromatin immunoprecipitation (ChIP) and ChIP-qPCR assays. Then, a mimic of miR-22, Nr3c1 plasmid encoding the glucocorticoid receptor (GR), and si-Nr3c1 were used to transfect AR42J cells, respectively. The mRNA expression of miR-22, Nr3c1, and Erb-b2 receptor tyrosine kinase 3 (ErbB3) was confirmed by qRT-PCR and the apoptosis rate of AR42J cells was detected by flow cytometry analysis. Western blot was used to detect the expression of ErbB3, GR, PI3k, PI3k-p85α, Akt, p-Akt, Bad, Bax, Bcl-xl, Bcl-2, and cleaved caspase3. RESULTS After inducing apoptosis of AR42J cells in vitro, the expression of miR-22 was significantly increased by 2.20 ± 0.26 and 4.19 ± 0.54 times, respectively, at 3 h and 6 h in comparison with the control group. As revealed by qRT-PCR assay, the expression of miR-22 was 78.25 ± 6.61 times higher in the miR-22 mimic group relative to the miRNA control group, accompanied with an obviously increased acinar cell apoptosis rate (32.53 ± 1.15 vs 18.07 ± 0.89, P = 0.0006). The upregulation of miR-22 could suppress its target gene, ErbB3, and the phosphorylation of PI3k and Akt. Furthermore, we predicted the potential transcription promoter of miR-22 and the binding sites using online tools. Luciferase reporter analysis and site-directed mutagenesis indicated that the binding site (GACAGCCATGTACA) of the GR, which is encoded by the Nr3c1 gene. Downregulation of the expression of GR could upregulate the expression of miR-22, which further promoted the apoptosis of AR42J cells. CONCLUSION GR transcriptionally represses the expression of miR-22, which further promotes the apoptosis of pancreatic acinar cells by downregulating the downstream signaling pathway.