Background: Previous studies reported that N-myc downstream-regulated gene 1 (NDRG1) was upregulated in various cancer tissues and decreased expression of miR-188-3p and miR-133b could suppress cell proliferation, metastasis, and invasion and induce apoptosis of cancer cells. However, the molecular mechanism of NRDG1 involved in hepatocellular carcinoma (HCC) tumorigenesis is still unknown. Methods: The expressions of miR-188-3p, miR-133b, and NRDG1 in HCC tissues and cells were quantified by qRT-PCR and Western blot. MTT assay and transwell invasion assay were performed to evaluate cell growth and cell migration, respectively. Luciferase reporter assay were performed to determine whether miR-188-3p and miR-133b could directly bind to NRDG1 in HCC cells. Results: The results showed that NRDG1 was upregulated and these 2 microRNAs were downregulated in HCC tissues. NRDG1 was negatively correlated with miR-188-3p and miR-133b in HCC tissues. MiR-188-3p and miR-133b were demonstrated to directly bind to 3′UTR of NRDG1 and inhibit its expression. Upregulation of miR-188-3p and miR-133b reduced NRDG1 expression in hepatocellular carcinoma cell lines, which consequently inhibited cell growth and cell migration. Conclusions: Our finding suggested that miR-188-3p and miR-133b exert a suppressive effect on hepatocellular carcinoma proliferation, invasion, and migration through downregulation of NDRG1.

ANXA1, which can bind phospholipid in a calcium dependent manner, is reported to play a pivotal role in tumor progression. However, the role and mechanism of ANXA1 involved in the occurrence and development of malignant glioma are still not well studied. Therefore, we explored the effects of ANXA1 on normal astrocytes and glioma cell proliferation, apoptosis, migration and invasion and the underlying mechanisms. We found that ANXA1 was markedly up-regulated in glioma cell lines and glioma tissues. Down-regulation of ANXA1 inhibited normal astrocytes and glioma cell proliferation and induced the cell apoptosis, which suggested that the consequences of loss of Annexin 1 are not specific to the tumor cells. Furthermore, the siRNA-ANXA1 treatment significantly reduced tumor growth rate and tumor weight. Moreover, decreasing ANXA1 expression caused G2/M phase arrest by repressing expression levels of cdc25C, cdc2 and cyclin B1. Interestingly, ANXA1 did not affect the expressions of β-catenin, GSK-3β and NF-κB, the key signaling molecules associated with cancer progression. However, siRNA-ANXA1 was found to negatively regulate phosphorylation of AKT and the expression and activity of MMP2/-9. Finally, the decrease of cell proliferation and invasiveness induced by ANXA1 down-regulation was partially reversed by combined treatment with AKT agonist insulin-like growth factor-1 (IGF-1). Meanwhile, the inhibition of glioma cell proliferation and invasiveness induced by ANXA1 down-regulation was further enhanced by combined treatment with AKT inhibitor LY294002. In summary, these findings demonstrate that ANXA1 regulates proliferation, migration and invasion of glioma cells via PI3K/AKT signaling pathway.

Objective:To investigate the effect of microRNA (miRNA, miR)-188-3p on N-myc downstream regulated gene 1(NDRG1) and its role in the proliferation activity of hepatocarcinoma cells (HCC).Methods:Collected 21 cases of HCC tissue specimens and 13 cases of adjacent cancer tissues that were surgically resected and pathologically confirmed by the Department of Hepatobiliary and Pancreatic Surgery, Wuhan Central Hospital from February 2018 to October 2019, HepG2 cells were cultured in vitro. In this study, the miR-188-3p was predicted with bioinformatics and identified with dual luciferase report system. Expression of miR-188-3p and NDRG1 was determined with real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting after transfection of miR-188-3p. The proliferation of HepG2 cells was detected in vitro by CCK-8 assay. Results:MiR-188-3p can inhibit NDRG1 expression by binding to 3’untranslated regions (3’UTR) of NDRG1 mRNA. miR-188-3p can negatively control the mRNA and protein expression of NDRG1 (0.56±0.04 vs.1.12±0.13, t=10.777, P<0.01、0.43±0.07 vs. 0.88±0.15, t=9.123, P<0.05). The proliferation of HepG2 cells was negatively controlled by miR-188-3p(0.76±0.07 vs. 1.26±0.19, t=7.509, P<0.05). Conclusion:MiR-188-3p could effectively inhibit the proliferation of HCC through targting NDRG1, suggesting miR-188-3p could serve as an innovative therapeutic strategy in terms of HCC.

目的 了解中小学教师骨质疏松症的患病率和影响因素,为针对性采取预防措施提供参考.方法 检测1 075例武汉市江岸区中小学教师的骨密度,采用SPSS 16.0软件包进行统计分析.结果 男性中小学教师身高、体重、BMI均明显高于女性(P＜0.05);40～49年龄组与50岁及以上年龄组男女中小学教师脚跟骨、腰椎总骨密度均明显低于29岁及以下年龄组(P＜0.05),50岁及以上年龄组男女中小学教师脚跟骨、腰椎总骨密度明显低于30～39年龄组(P＜0.05);随着男女中小学教师年龄逐渐增长,骨量流失和骨质疏松症发生率明显升高(P＜0.05),且女性中小学教师骨量流失和骨质疏松症发生率明显高于男性中小学教师(P＜0.05);经多因素Logistic回归分析发现,年龄为脚跟骨、腰椎总骨密度的负相关因素,身高、体重、BMI为脚跟骨、腰椎总骨密度的正相关因素.结论 随着年龄增长,中小学教师脚跟骨、腰椎总骨密度明显下降,骨量流失和骨质疏松症发生率明显升高,且女性表现尤为明显,建议采取适宜运动锻炼和合理饮食配伍等多方面综合措施,早期防治骨质疏松症的发生.

Objective To evaluate the effect of Annexin-A1 mimetic peptide Ac2-26 on activation of astrocytes in rats.Methods The primarily cultured astrocytes from the cortex of fetal Sprague-Dawley rats after 4 passages were divined into 4 groups (n =24 each) using a random number table method:control group (C group),LPS group,LPS+scramble peptide group (LPS+Src group) and LPS+Ac2-26 group.LPS was added to LPS group with the final concentration of 1 mg/ml.LPS at the final concentration of 1 mg/ml and scramble peptide at the final concentration of 3.3 mmol/L were added to LPS+Src group.LPS at the final concentration of 1 mg/ml and Ac2-26 at the final concentration of 3.3 mmol/L were added to LPS+Ac2-26 group.After 24-h incubation,the cell survival rate was measured by CCK-8 assay,the migration was determined by Transwell assay,the concentrations of tumor necrosis factor-alpha (TNF-α),interleukin-1 beta (IL-1β),monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-1 a (MIP-1a) in the supernatant were measured (by enzyme-linked immunosorbent assay),and the expression of glial fibrillary acidic protein (GFAP),extracellular signal-regulated kinase (ERK),phosphorylated ERK (p-ERK),c-Jun N-terminal kinase (JNK),phosphorylated JNK (p-JNK),p38 mitogen-activated protein kinase (p38MAPK),and phosphorylated p38MAPK (p-p38MAPK) in astrocytes was detected by Western blot.Results Compared with group C,the expression of GFAP was significantly up-regulated,and the cell mobility,concentrations of TNF-α,IL-1β,MCP-1 and MIP-1α in the supernatant,p-ERK/ERK ratio,p-JNK/JNK ratio and p-p38MAPK/p38MAPK ratio were increased (P＜0.05),and no siguificant change was found in the cell survival rate in group LPS (P＞0.05).Compared with group LPS,the expression of GFAP was significantly down-regulated,and the cell mobility,concentrations of TNF-α,IL-1β,MCP-1 and MIP-1α in the supernatant,p-JNK/JNK ratio and p-p38MAPK/p38MAPK ratio were decreased in group LPS+Ac2-26 (P＜0.05),and no significant change was found in the parameters mentioned above in group LPS+Src (P＞0.05).Conclusion Ac2-26 can inhibit activation of astrocytes and produces anti-inflammatory effect in rats.

Objective To explore the expression and clinical significance of the long non-coding RNA anti-differentiation antagonizing noncoding RNA (DANCR) in the patients with intrahepatic metastasis of hepatocellular carcinoma (HCC).Methods The tumor tissues,pre-and post-operative plasma samples of 24 HCC intrahepatic metastasis patients undergoing resection,the tumor tissues and plasma samples of 24 HCC patients,and the plasma samples of 24 healthy volunteers were collected from the Central Hospital of Wuhan between Jan.2015 and Oct.2017.The expression of DANCR mRNA level was detected by quantitative real-time quantitative polymerase chain reaction (PCR) in the tumor tissues and plasma samples.The relationship of DANCR mRNA expression with clinicopathological characteristics was analyzed.Results The expression levels of DANCR mRNA in the tumor tissues (2.85 ± 0.23 vs.1.63 ±0.16,P＜0.01) and the plasma (0.043 ±0.007 vs.0.018 ±0.005,P＜0.01) of the patients with intrahepatic metastasis of HCC were significantly higher than those of HCC patients.In addition,the DANCR mRNA expression was significantly lower after surgical resection than that in patients with intrahepatic metastasis of HCC before surgery (0.035 ±0.006 vs.0.012 ±0.004,P ＜0.05).The expression of DANCR mRNA in tumor tissues of patients with HCC intrahepatic metastasis was correlated with the size of primary tumor (P ＜ 0.05),chronic hepatitis B history (P ＜ 0.05) and Child-Pugh grade (P ＜ 0.05).Conclusion DANCR expression is closely related to the development of HCC.

Overexpression of long noncoding RNA (lncRNA) H19 has been observed in various cancers, which indicates that H19 exert important roles in the progression of carcinogenesis. MiR-326 has been reported to play tumor suppressive roles in multiple tumors. Recently, the competing endogenous RNA (ceRNA) hypothesis has implied that lncRNAs might function as molecular sponges for microRNAs in various cancers. However, the roles of H19/miR-326 in human hepatocellular carcinoma (HCC) still remain unclear. The aim of our study was to determine H19/miR-326 expression in HCC cells and investigate their roles in HCC development. We found that H19 was significantly elevated and miR-326 was decreased in HCC cells including Hep3B, HepG2, MHCC-97L, SK-hep1, Hun7, SMCC-7721 compared with LO2 cells, respectively. In the subsequent experiments, we observed that inhibition of H19 can repress HCC cell growth, migration, and invasion in vitro. H19 downregulation can increase miR-326 expression in HCC cells. Meanwhile, miR-326 mimics can also inhibit HCC progression, whereas miR-326 inhibitors exhibited a reverse phenomenon by modulating H19 expression. In addition, a negative association between H19 and miR-326 was predicted and confirmed. Furthermore, the transcription factor TWIST1 has been recognized as a significant regulator in tumor progression. Here, by performing bioinformatics analysis, TWIST1 was identified as a downstream target of miR-326. The findings of our study implied that lncRNA H19 can serve as a ceRNA to sponge miR-326 and modulate TWIST1 levels in HCC pathogenesis. Taken these together, these findings indicated that H19/miR-326/TWIST1 axis was involved in HCC development and can indicate a novel HCC target.

Purpose: To further observe and verify the effect of nonalcoholic fatty liver disease (NAFLD) on the response to antiviral therapy in patients with chronic hepatitis B (CHB) and investigate the relationship between the virologic response and insulin resistance.Patients and methods: A retrospective study was adopted and 61 NAFLD patients with HBeAg-positive CHB were included as the observation group (group A), and 64 patients with simple CHB were included as the control group (group B).Results: After 12 weeks of treatment with entecavir, the total virologic response rate in group A was statistically significantly lower than that in group B (P<0.05). During weeks 24-96, the difference was not statistically significant (P>0.05). In weeks 48 and 96, there was no significant difference in the HBeAg seroconversion rates between the two groups (P>0.05). In weeks 12 and 24, there was also no significant difference in the alanine transaminase (ALT) normalization rate between the two groups (P>0.05). Then, in weeks 48 and 96, the ALT normalization rate of group A was obviously lower than that of group B (P<0.05). Group A patients were divided into group A1 (<= M) and group A2 (>M) according to the median value (M=2.79) of the baseline homeostatic model assessment method insulin resistance levels. In weeks 48 and 96, the ALT normalization rate of group A1 was significantly higher than that of group A2 (P<0.05). The correlation coefficient (r) of the baseline homeostatic model assessment method insulin resistance level and the severity of fatty liver in group A was 0.426 (P=0.001).Conclusion: NAFLD cannot affect the long-term total virologic response rate and HBeAg seroconversion rate in CHB patients treated with entecavir but can reduce the long-term biochemical response rate, which has a positive correlation with the severity of fatty liver and the insulin resistance index.

Objective To investigate the clinicopathologic significance of microRNA (miRNA,miR)-34a expression in hepatocellular carcinomas (HCC).Methods Real-time quantitative polymerasc chain reaction (Real-time PCR) was used to quantify miR-34a expression in 59 pairs of HCC tissues and matched normal hepatic tissues.The association of miR-34a expression with the clinicopathologic features was evaluated.Results The expression of miR-34a was significantly downregulated in HCC tissue samples (0.52 ±0.21) compared to matched normal tissues (5.57 ± 1.48,P ＜0.05).MiR-34a was highly expressed in HCC with a low pathological grade (Ⅰ-Ⅱ:0.69 ± 0.22) but markedly decreased in grade (Ⅲ-Ⅳ:0.32 ±0.12) tumors (P ＜0.01).In addition,there was significant difference in the expression of miR-34a between patients with TNM stages Ⅰ-Ⅱ (0.72 ± 0.19) and those with stage Ⅲ-Ⅳ (0.35 ± 0.13) HCC (P ＜ 0.01).Conclusion The expression of miR-34a is closely related to the development and progression of HCC.

Objective To investigate the effects of Annexin-A1 ( Anxa1 ) gene silencing induced by siRNA on the growth and migration of microglial BV-2 cells and its possible mechanisms.Methods A synthesized siRNA duplex targeting Anxa1 gene was transfected into BV-2 cells.The efficiency of siRNA-in-duced Anxa1 gene silencing was evaluated on both mRNA and protein levels by using reverse-transcription PCR and Western blot assay.MTT assay was performed to measure the proliferation of BV-2 cells with si-lenced expression of Anxa1 gene.Flow cytometry with Annexin V-FITC/PI double staining was used to de-tect the apoptosis rate of BV-2 cells.Transwell chambers were used to analyze the effects of siRNA-induced Anxa1 gene silencing on the migration of BV-2 cells.Western blot assay was performed to detect the expres-sion of signaling proteins related to cell cycle and migration.Results Compared with the siRNA negative control ( siRNA-NC) group, the inhibitory rates of siRNA-induced Anxa1 gene silencing on the proliferation of BV-2 cells were significantly increased at the time points of 24 h, 48 h and 72 h after intervention [(16.9 ±2.1)%, (23.1±3.6)%and (42.4±1.7)%vs (1.35±0.5)%, (2.06±0.7)% and (8.65±0.9)%, P<0.05 ].The apoptosis rate of BV-2 cells transfected with Anxa1 siRNA was (18.4±2.1)%, which was significantly elevated as compared with that of the siRNA-NC group (5.2±0.3)%and control group (4.3±0.2)%.Cell migration of the Anxa1 siRNA transfected BV-2 cells was inhibited remarkably at 48 h as com-pared with that of the siRNA-NC group (28.7±5.2 vs 173.4±11.4, P<0.01).Moreover, the suppressed expression of Cyclin D1 protein and activation of p38 and JNK signaling pathways were induced by silenced expression of Anxa1 gene in BV-2 cells.Conclusion The growth and migration of BV-2 cells were signifi-cantly inhibited by silencing the expression of Anxa1 gene with siRNA, the possible mechanisms might be associated with the suppressed expression of Cyclin D1protein and the activation of p38 and JNK signaling pathways.

目的：探讨miR－218与宫颈癌侵袭转移和不良预后因素之间的关系。方法用qRT－PCR的方法检测112例宫颈癌患者和21例正常对照的宫颈组织和外周血中miR－218的表达水平，所得数据结合临床资料进行相关分析。结果 miR－218在宫颈癌组织的表达水平明显低于宫颈正常上皮组织，差异有统计学意义（ P＜0.001）。 miR－218的表达水平与宫颈癌FIGO分期、淋巴结转移、深间质浸润、阴道壁累及、脉管浸润有关。结论miR－218在宫颈癌组织中表达下调，且与宫颈癌侵袭转移和不良预后因素密切相关，提示miR－218在宫颈癌的侵袭转移机制中发挥了重要作用。

Objective To investigate the effect of microRNA-34a (miR-34a) on X-linked inhibitor of apoptosis protein (XIAP) and its role in the proliferation activity and cell apoptosis of hepatocarcinoma cells.Methods Synthetic miR-34a mimic and its negative control were transfected into human hepatocarcinoma SMMC7721 cells to a final concentration of 200 nmol/L by liposome method.After transfection,the cell counting kit-8 (CCK-8) was used to measure the cell proliferation activity.The cell apoptosis was determined by flow cytometry.The expression of XIAP mRNA and protein was detected by real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) and Western blotting respectively.Results The transfection efficiency mediated by cationic liposome was greater than 85％.The inhibitory rate of cell proliferation in SMMC7721 cells was (39.5 ± 10.5)％,(43.5 ± 2.8)％ and (49.6 ± 0.3) ％ at 24,48 and 72 h respectively after transfection with miR-34a mimic,which was significantly increased as compared with the control cells.Moreover,the cell apoptosis in experimental group (6.98 ± 0.13)％ was higher than that in negative control group (4.89 ± 0.62)％ and simple liposome group (4.02±0.21)％ at 72 h.Whereas the expression of XIAP protein was decreased in SMMC7721 cells transfected with miR-34a mimic at 72 h after transfection.Conclusion miR-34a suppresses the proliferation activity of hepatocarcinoma cells by down-regulating XIAP gene expression.

>胃癌是常见的恶性肿瘤之一,其预后尚不理想。DKK-1(dickkopf-1)是Wnt信号通路的一种拮抗剂,但DKK-1在胃癌发生中的生物学效应尚未阐明,研究旨在通过检测胃癌患者血清及组织中DKK-1的表达,探讨其在胃癌诊断中的意义。一、资料与方法1.对象:选取本院2012年3月至2013年10月,拟手术切除胃癌患者78例,术前未接受放、化疗。男50例,女28

目的 探讨胰腺癌组织中表皮生长因子受体(EGFR)、基质金属蛋白酶9(MMP-9)的表达及其与胰腺癌临床病理特征的关系.方法 应用免疫组织化学SP法检测44例胰腺癌、相应癌旁胰腺组织、13例慢性胰腺炎和7例正常胰腺组织中EGFR、MMP-9的表达,并分析其与临床病理特征的相关性.结果 正常胰腺组织中均无EGFR及MMP-9的表达.慢性胰腺炎组织中EGFR、MMP-9的阳性表达率分别为23.1％(3/13)和15.4％(2/13).胰腺癌组织中EGFR、MMP-9的阳性表达率分别为61.4％(27/44)和54.5％(24/44);相应癌旁组织中的阳性表达率分别为34.1％(15/44)和29.5％(13/44).胰腺癌组织中的EGFR及MMP-9阳性表达率均显著高于相应癌旁组织(P＜0.05);胰腺癌组织及癌旁组织中EGFR及MMP-9的表达均显著高于慢性胰腺炎组织和正常胰腺组织(P均＜0.05).EGFR和MMP-9的表达与胰腺癌临床分期、分化程度、血管侵犯及淋巴结转移相关.结论 胰腺癌组织中EGFR的表达与MMP-9表达密切相关,EGFR的表达与MMP-9的表达水平可作为了解胰腺癌生物学行为和判断预后的指标.

Objective To investigate the dysregulation of microRNA-500 (miR-500) in the development process from hepatitis C liver cirrhosis to hepatocellular carcinoma so as to clarify the role of change of miR-500 expression in the development process from the hepatitis C to liver cirrhosis and hepatocellular carcinoma.Methods From Nov.2012 to Oct.2013,8 cases of hepatitis C liver cirrhosis and 8 cases of hepatitis C virus infection-related liver cancer were selected as the research objects,and 8 cases of the healthy check-up served as the control group.The expression level of miR-500 in the plasma of the above three groups was tested by using real-time quantitative polymerase chain reaction (Real-time PCR).Results The expression level of miR-500 in liver cirrhosis group was 5.72 ± 2.28,higher than 0.95 ± 0.48 in the control group,and that in liver cancer group was 18.62 ± 8.13,higher than in the control group and liver cirrhosis group.Conclusion The miR-500 may be involved in the development process from chronic hepatitis C liver cirrhosis to liver cancer.

Objective To explore the role of microRNA-181b (miR-181b) in the pathogenesis of human hepatocellular carcinoma (HCC).Methods We detected the expressions of miR-181 b by using real-time quantitative polymerase chain reaction (Real-time PCR) in LO2 (normal human liver cell line),HepG2 and SMMC7721 (human HCC cell lines),normal liver tissues,para-cancerous tissue and cancerous tissues surgically resected from 35 cases of HCC.Cell Counting Kit-8 (CCK-8) assay and growth curve were used to asses the effect of miR-181b inhibitor on proliferation of HCC cells.Results The expression of miR-181b in HepG2 and SMMC7721 cells was significantly higher than in LO2 cells (0.582 ± 0.032 and 0.716 ±0.045 vs.0.255 ±0.039,P ＜0.05),and that in cancerous tissue was significantly higher than in normal liver tissues and para-cancerous tissues (0.636 ± 0.038 vs.0.156-± 0.005 and 0.167 ± 0.023,P ＜0.05).miR-181b inhibitor could reduce the proliferative ability of HCC cells.Conclusion These results indicate that the high expression of miR-181b is probably involved in HCC pathogenesis.Downregulation of miR-181b expression can inhibit the proliferation of HCC cells.miR-181b may act as a molecular tar-get for HCC diagnosis and targeted therapy.

Objective To investigate the regulatory effect ofmicroRNA-181b (miR-181b) on deleted in liver cancer-1 (DLC-1) and its role in hepatocarcinogenesis.Methods miR-181b and DLC-1 mRNA expression levels in normal tissue,paraneoplastic tissue,liver cancer tissue and liver cancer cells were detected by using real-time fluorescent quantitative polymerase chain reaction (FQ-PCR).DLC-1 protein expression was detected by using Western blotting.miR-181b inhibitor was transfected into SMMC7721 cells and the expression changes of DLC-1 mRNA and protein were examined by using FQ-PCR and Western blotting respectively.Results miR-181b was up-regulated in hepatocellular carcinoma and adjacent tissues as compared to normal liver tissues and the expression of miR-181 b was also increased in SMMC7721 cells as compared to fetal liver cells LO2,whereas the expression levels of DLC-1 mRNA and protein were decreased in paraneoplastic and HCC tissues compared with normal liver tissues.The expression of miR-181b and DLC-1 was negatively correlated.No obvious difference was found in the DLC-1 mRNA expression after miR-181b inhibitor transfection.Conclusion The DLC-1 abnormal low-expression may play a dynamic role in hepatocarcinogenesis due to the miR-181 b up-regulation.

目的探讨基质金属蛋白酶-9(MMP-9)在前列腺癌发生、发展中的作用。方法采用RT-PCR法检测MMP-9在52份前列腺癌和18份前列腺增生(BPH)组织中的表达,分析其与前列腺癌临床病理特征的关系。结果前列腺癌组织中MMP-9的表达明显高于BPH组织,P<0.01。MMP-9表达随着前列腺癌分化程度的降低而升高,随临床分期的增高而升高,P均<0.05。结论 MMP-9过表达可促进前列腺癌的发生与发展;其水平变化可为前列腺癌的临床诊断、治疗及预后判断提供依据。