Effects of oxymatrine on the expression and release of high mobility group box 1 in pancreatic acinar cells stimulated by hydrogen peroxide / 中华胰腺病杂志

Objective@#To observe the effects of oxymatrine(OM) on the expression and release of high mobility group box 1(HMGB1) in rat pancreatic acinar cell line AR42J stimulated by hydrogen peroxide.@*Methods@#MTT method was used to detect the effects of H2O2 in different concentrations on the survival of AR42J cells. AR42J cells cultured in vitro were divided into control group, H2O2 group and H2O2+ OM group. An equal volume of H2O2(final concentration 0.16 mmol/L) was added in H2O2 group and H2O2+ OM group, respectively, while an equal volume of triple distilled water was added in control group. In H2O2+ OM group, OM(final concentration 0.5 g/L)was added 0.5 h before the addition of H2O2, and cell samples and supernatant were collected after 24 h culture. The expression of HMGB1 protein was detected by Western blotting, the level of HMGB1 protein in cell supernatant was detected by enzyme-linked immunosorbent assay, and the intracellular distribution of HMGB1 protein was detected by immunofluorescence.@*Results@#In the H2O2 group, the expression of HMGB1, the secretion of HMGB1 in the supernatant and the proportion of cytoplasmic HMGB1 in the total HMGB1 were significantly higher than those in the control group[1.04±0.04 vs 0.69±0.02, (4.84±0.13)μg/L vs (2.68±0.07)μg/L, (35.7±2.5)% vs (10.7±1.9)%], and all the differences were statistically significant (all P<0.05). After OM intervention, HMGB1 protein expression, secretion and cytoplasmic proportion were [0.82±0.02, (3.97±0.10)μg/L and (27.3±1.7)%], respectively, which were obviously lower than those in the H2O2 group, and all the differences were statistically significant (all P<0.05).@*Conclusions@#H2O2 can induce the expression and release of HMGB1 in rat pancreatic acinar cells; OM treatment could alleviate the severity of oxidative stress injuries induced by H2O2 in AR42J cells.

Expression and clinical significance of CCDC34 in hepatocellular carcinoma: An analysis based on The Cancer Genome Atlas database / 临床肝胆病杂志

ObjectiveTo investigate the expression and clinical value of coiled-coil domain-containing protein 34 (CCDC34) in hepatocellular carcinoma (HCC), and to predict the role of CCDC34 in the development and progression of HCC. MethodsThe datasets of HCC were downloaded from The Cancer Genome Atlas (TCGA) to obtain the expression profile and clinical information of the CCDC34 gene. The bioinformatics method was used to analyze the expression of CCDC34 in HCC, its correlation with clinicopathological parameters, and its influence on prognosis. The gene set enrichment analysis (GSEA) was used to predict the possible pathways regulated by the CCDC34 gene in HCC. The independent samples t-test and the paired t-test were used for comparison of continuous data between two groups; the Kaplan-Meier method and the log-rank test were used for survival analysis; the Cox proportional-hazards regression model analysis was used to investigate the influencing factors for prognosis. P＜0.01 was the standard for judging significant enrichment in GSEA, and the false discovery rate was ＜0.05. ResultsIn TCGA database, CCDC34 was highly expressed in tumor tissue, and there was a significant difference in the expression of CCDC34 between patients with different TNM stages and tumor grades (t=2.118 and 3.622, P=0.035 and P＜0.001). The patients with high expression of CCDC34 had a significantly shorter overall survival time than those with low expression (χ2=21.716, P＜005). The multivariate Cox regression analysis showed that the expression of CCDC34 （HR＝2.287,95%CI:1.312-3.987）and TNM stage(HR=1.943,95%CI:1.101-3.429) were independent risk factors for the overall survival time of patients with HCC (all P＜0.05). The enrichment of 8 pathway gene sets, including base excision repair and spliceosome, was observed in the samples with high expression of CCDC34 (P＜0.01, FDR ＜0.05). ConclusionCCDC34 may play a vital role in the development and progression of HCC and thus become a new prognostic indicator and a potential therapeutic target.

Expression and clinical significance of centromere protein A in pancreatic cancer based on GEO and TCGA database analysis / 中华胰腺病杂志

Objective:To study the expression and clinical value of centromere protein A (CENP-A) in pancreatic cancer (PC), and to predict the role of CENP-A in the development and progression of PC.Methods:The expression profile of CENP-A mRNA and clinicopathological data were obtained from PC data set downloaded from the gene expression omnibus (GEO) and the cancer genome atlas (TCGA), which included a microarray data set of 45 patients with PC (GSE28735) and a microarray data set of 177 patients with PC. The correlation between CENP-A gene expression in PC tissue and the clinicopathological indices as well as the effects on the prognosis of PC was analyzed by the bioinformatics method. The potential pathway regulated by CENP-A in PC was predicted by gene set enrichment analysis (GSEA). Meanwhile, the proteins related to CENP-A were screened out by combining with the protein-protein interaction (PPI) network of CENP-A in STRING database, and the correlation between CENP-A and expression of its related proteins was analyzed.Results:The expression level of CENP-A mRNA in PC tissue was greatly higher than that in adjacent tissue, and the difference was statistically significant (4.492±0.361 vs 3.431±0.405, P=0.001). The expression level of CENP-A in PC tissue was significantly correlated with TNM stage and tumor grade ( P<0.05). The overall survival of patients with high CENP-A expression was significantly lower than that of patients with low CENP-A expression ( P<0.05). Cell cycle and DNA replication pathways were enriched in PC tissue with high CENP-A expression ( P<0.01, false discovery rate<0.05). CENP-A in PC tissue was strongly correlated with CDK1 ( r=0.861, P<0.001) and BUB1 ( r=0.825, P<0.001) in PC tissue through the PPI network analysis. Conclusions:CENP-A gene overexpression in PC tissue was related to its malignant biological behaviour, and its mechanism may be achieved by the synergistic action with CDK1 and BUB1.

Effects of oxymatrine on the expression and release of high mobility group box 1 in pancreatic acinar cells stimulated by hydrogen peroxide / 中华胰腺病杂志

Objective:To observe the effects of oxymatrine(OM) on the expression and release of high mobility group box 1(HMGB1) in rat pancreatic acinar cell line AR42J stimulated by hydrogen peroxide.Methods:MTT method was used to detect the effects of H 2O 2 in different concentrations on the survival of AR42J cells. AR42J cells cultured in vitro were divided into control group, H 2O 2 group and H 2O 2+ OM group. An equal volume of H 2O 2(final concentration 0.16 mmol/L) was added in H 2O 2 group and H 2O 2+ OM group, respectively, while an equal volume of triple distilled water was added in control group. In H 2O 2+ OM group, OM(final concentration 0.5 g/L)was added 0.5 h before the addition of H 2O 2, and cell samples and supernatant were collected after 24 h culture. The expression of HMGB1 protein was detected by Western blotting, the level of HMGB1 protein in cell supernatant was detected by enzyme-linked immunosorbent assay, and the intracellular distribution of HMGB1 protein was detected by immunofluorescence. Results:In the H 2O 2 group, the expression of HMGB1, the secretion of HMGB1 in the supernatant and the proportion of cytoplasmic HMGB1 in the total HMGB1 were significantly higher than those in the control group[1.04±0.04 vs 0.69±0.02, (4.84±0.13)μg/L vs (2.68±0.07)μg/L, (35.7±2.5)% vs (10.7±1.9)%], and all the differences were statistically significant (all P<0.05). After OM intervention, HMGB1 protein expression, secretion and cytoplasmic proportion were [0.82±0.02, (3.97±0.10)μg/L and (27.3±1.7)%], respectively, which were obviously lower than those in the H 2O 2 group, and all the differences were statistically significant (all P<0.05). Conclusions:H 2O 2 can induce the expression and release of HMGB1 in rat pancreatic acinar cells; OM treatment could alleviate the severity of oxidative stress injuries induced by H 2O 2 in AR42J cells.

Research progress of TLR4 signaling pathway related miRNAs in inflammatory response / 天津医药

Toll-like receptor 4 (TLR4) plays an important role in inflammation and immune response.MicroRNAs (miRNAs) are involved in the regulation of TLR4 signaling pathway in multiple levels and various molecules,which play an important role in inflammatory reaction.A variety of miRNAs are involved in the regulation of TLR4 signaling pathway,and the TLR4 signaling pathway can induce a variety of miRNAs.Chronic diseases such as diabetes,Alzheimer's disease and cardiovascular disease are closely related to inflammatory response.The regulatory role of TLR4 signaling related miRNAs has attracted much attention in inflammatory diseases.In this review,the research progress of TLR4 signaling pathway related miRNAs in the regulation of inflammatory response is summarized,which provides a new research direction for the clinical diagnosis and treatment of inflammatory response related diseases.

The influence of LPS on the protein expression of related molecules in Smads and ERK1/2 signal pathway in LTC-14 cells / 中华胰腺病杂志

Objective To explore the influence of LPS treatment on related molecules in Smads and ERK1/2 signal pathway in pancreatic stellate cell line LTC-14.Methods LTC-14 cells were cultured in vitro, and were treated with LPS at different dose in different time points.Protein expressions of related molecules in Smads pathway and ERK1/2 pathway and α-SMA in LTC-14 Cells were examined by Western blot.Results On Treated LTC-14 cells by 0, 1, 5, 10, 20 and 50 mg/L LPS,protein expressions of Smad3 were 0.15±0.02, 0.37±0.02, 0.44±0.01, 0.46±0.02, 0.372±0.01 and 0.24±0.03;expressions of Smad7 were 0.79±0.05, 0.84±0.02, 0.55±0.03, 0.45±0.03, 0.34±0.02 and 0.92±0.07;p-ERK1/2 levels were 0.48±0.05, 0.74±0.03, 0.72±0.04, 0.89±0.02, 0.81±0.02 and 0.72±0.03;p-cPLA2 levels were 0.15±0.03, 0.30±0.01, 0.31±0.01, 0.30±0.02, 0.28±0.03 and 0.32±0.02;α-SMA levels were 0.56±0.06, 0.62±0.06, 0.54±0.04, 1.03±0.11, 1.39±0.08 and 1.28±0.10.The changes of protein expressions before and after LPS treatment were obvious (all P<0.01).The protein expressions of ERK1/2 were 0.56±0.03, 0.57±0.02, 0.53±0.02, 0.58±0.02, 0.59±0.05 and 0.55±0.04, which did not change obviously along with increased LPS dosages.LTC-14 cells treated with 10 mg/L LPS for 0, 1, 3, 6 and 9 h,the expressions of Smad3 were 0.69±0.05, 0.68±0.07, 1.02±0.14, 1.82±0.0 and 2.04±0.11,those of Smad7 were 2.77±0.10, 1.37±0.08, 1.45±0.14, 0.78±0.09 and 0.63±0.06,those of p-ERK1/2 were 0.16±0.03, 0.32±0.05, 0.79±0.03, 1.50±0.07 and 1.77±0.04,those of p-cPLA2 were 0.15±0.04, 0.32±0.06, 0.63±0.04, 0.95±0.04 and 1.49±0.10,those of α-SMA were 0.84±0.03, 1.26±0.21, 1.81±0.19, 4.28±0.26 and 4.37±0.15, all of which changed obviously as the treatment time increased (P<0.05 or 0.01).The expressions of ERK1/2 were 0.75±0.03, 0.72±0.02, 0.80±0.04, 0.74±0.03 and 0.85±0.09, which did not change obviously as the treatment time increased.Conclusions LPS could upregulate the expression of α-SMA in a time-and dose-dependent way, and activate intracellular Smads and ERK1/2 inflammatory pathways, which may be the potential molecular mechanism of the development of chronic pancreatitis.

Effects of all-trans retinoic acid on sensitivity of gastric cancer cells to ra-diotherapy / 中国病理生理杂志

AIM:To investigate the effect of all-trans retinoic acid ( ATRA) on the viability of gastric cancer cell SGC-7901 and the sensitivity to radiotherapy .METHODS:MTT assay was used to examine the cell viability .Radio-sensitivity and cell cycle were determined by colony formation assay and flow cytometry , respectively .The mRNA levels of Bax, Bcl-2, survivin and NF-κB in the cells were measured by RT-qPCR.RESULTS: ATRA inhibited the viability of SGC-7901 cells in a concentration-dependent manner .The maximal inhibition was at concentration of 8 μmol/L.Colony formation assay revealed that the combination of ATRA with X-ray treatment significantly reduced the values of D 0 and Dq, and shifted down the fitting survival curve , as compared with radiotherapy alone .Moreover , ATAR markedly decreased the percentage of G2/M phase in the SGC-7901 cells (P<0.05).In addition, following ATRA treatment, the mRNA levels of Bcl-2 and survivin were decreased (P<0.05), whereas the mRNA levels of Bax and NF-κB were increased (P<0.05). CONCLUSION:ATRA enhances the sensitivity of SGC-7901 cells to radiotherapy , inhibits G2/M arrest and regulates the mRNA expression of Bax , Bcl-2, survivin and NF-κB.

Oxymatrine suppressed the activation of pancreatic stellate cells and p38-MAPK mRNA expression in vitro / 中华胰腺病杂志

Objective To clarify whether oxymatrine ( OM) could suppress the activation of pancreatic stellate cells ( PSC) and explore the potential molecular mechanism .Methods The proliferation of PSC line LTC 14 being activated by TGF-β1 with OM treatment at different concentrations (OM group) was measured. SOD level was determined by ELISA and p 38-MAPK mRNA was determined by real-time PCR.Results The proliferation of PSC in the control group , 0.1, 0.5, 1, 2, 5 g/L OM group was (1.51 ±0.08), (1.50 ± 0.07), (1.15 ±0.04), (1.15 ±0.04), (1.08 ±0.06), and (1.08 ±0.10), respectively.The level of the control group was lower than the groups where the concentration of OM reached or exceeded 0.5mg/ml ( all P=0.000).SOD level of LTC 14 cells in the control group, TGF-β1 group, 0.5 and 1 g/L OM group was (0.087 ±0.005), (0.073 ± 0.004), (0.085 ± 0.010), and (0.086 ± 0.007), respectively. No statistically significant difference existed among the groups (P=0.095).The p38-MAPK mRNA expression of PSC in the control group, TGF-β1 group, 0.5, and 1 g/L OM group was (1.000 ±0.000), (1.979 ± 0.505), (0.606 ±0.111), and (0.303 ±0.159), respectively.The p38-MAPK mRNA level of TGF-β1 group was higher than that of the control group (P=0.002), and that of 0.5 mg/ml OM group and 1 mg/ml OM group was lower that of TGF-β1 group ( P=0.000 ) , while no statistical difference was found between 0.5 mg/ml OM group and 1 mg/ml OM group.Conclusions OM could suppress the activation of PSC in vitro and the suppression of p38-MAPK mRNA expression may be involved .

Blockade of CLC-3 chloride channel inhibited the viability and invasion of colorectal cancer cells / 中国肿瘤临床

Objective:To examine the expression of CLC-3 in colorectal tissues and the effect of CLC-3 on the viability and invasion of colorectal cancer (CRC) SW480 and SW620 cells. Methods:The mRNA levels of CLC-3 in CRC cell lines were determined by RT-PCR. CLC-3 expression was inhibited by adding DIDS or NPPB to the CRC cells. Subsequently, cell viability and invasion were assessed by CCK-8 assay and Transwell assay, respectively. In addition, the effects of DIDS and NPPB on the Wnt orβ-catenin signaling pathways were de-termined by Western blot analysis. Results:The mRNA level of CLC-3 was remarkably increased in the CRC tissues compared with that in normal colorectal tissues (P<0.05) and was positively correlated with the T stage of CRC. The blockade of CLC-3 inhibited the viability and invasion of CRC cells (P<0.05). The expression ofβ-catenin, C-myc, cyclin D1, Ki-67, and survivin were evidently reduced by the in-hibition of CLC-3 (P<0.05). Conclusion:The inhibition of CLC-3 decreases the cell viability and invasion of CRC cells by reducing the ex-pression of the proteins related to the Wnt orβ-catenin signaling pathway.

STAT3 signaling in pathogenesis of pancreatitis / 中国病理生理杂志

Signal transducer and activator of transcription 3 (STAT3), an acute-phase response protein, is ac-tivated to over-express by cytokines .STAT3 also acts as a transcriptional factor to regulate the expression of cytokines .O-ver-expression of cytokines is accompanied by STAT 3 activation and over-expression in acute pancreatitis .Meanwhile , the proliferation of pancreatic stellate cells in chronic pancreatitis is mediated by STAT 3.In this review, the research progress in STAT3 function is summarized to elaborate its potential role in the pathogenesis of pancreatitis .

Design of artifact-free head holder for small animal imaging / 中国医学装备

Objective:To design a kind of head holder for small animal imaging without artifact. Methods: The head holder was designed for small animal imaging according to rodent head stereotaxic apparatus with lightweight and easy molding material (polymethyl methacrylate). The holder can be easily made with ordinary lathe and can be easily operated in small animal imaging device. Results:After the trial in small animal PET-CT, the holder can effectively solve the problems in small animal imaging such as the artifact caused by tiny displacement, tilt head position, deviating from the central field of view and respiratory depression. Conclusion: The small animal imaging device was designed in simple structure to overcome the shortcomings of the existing technology. The operation of the device is simple, which is worthy of popularization head holder of application.