Objective To study the expression of ubiquitin-activating enzyme E1 (UBE1) and Bct-2 in diffuse large B cell lymphoma (DLBCL) and its clinicopathological relevance. Methods Tumor tissues were collected from 80 DLBCL patients, and tissues from 30 patientswith reactive lymphoid hyperplasia (RH) were taken as controls. Immunohistochemistry detection (EnVision method) was used to detect the expressions of CD10, Bct-6 and MUM1 inDLBCL tissues for immunology typing of GCB or ABC. The expression of UBE1 and Bct-2 proteins in DLBCL tissues and RH tissues was assayed by immunohistochemical staining, and its correlation with patients' clinical pathological indices were analyzed. We also analyzed the correlation of UBE1 expression with Bct-2 expression in DLBCL tissues. Results The positive rates of UBE1 were 65. 0% (52/80) in DLBCL tissues and 13. 3%(4/30) in RH tissues (P<0. 05). The positive rates of UBE1 were 53. 3% (16/30) for GCB type DLBCL and 72. 0%(36/50) for ABC type (P<0. 05). The positive rates of Bct-2 were 51. 3% (41/80) in DLBCL tissues and 10. 0% (3/30) in RH tissues (P<0. 05). The positive rates of Bcl-2 were 36. 7%(11/30) for GCB type DLBCL and 60. 0% (30/50) for ABC type (P<0. 05). The positive rate of UBE1 was correlated with age and the extranodal involvement site (P<0. 05). The positiverate of Bcl-2 was correlated with age, Ann Arborstage, extranodal involvement site and international prognostic index (P<0. 05). UBE1expression was positively correlated with Bcl-2 in ABC type DLBCL (r =0.582, P = 0. 001). Conclusion UBE1 and Bcl-2 proteins are highly expressed in ABC type DLBCL tissues, which probably plays a role in the development and progression of DLBCL. High expression of UBE1 and Bcl-2 may indicate poor prognosis of DLBCL patients.

AIM: To investigate the effects of agkis-trodon halys venom anti-tumor component (AHVAC-) on the biological behavior of gastric cancer MKN-28 cells. METHODS: Gastric cancer MKN-28 cells were treated with the experimental concentrations (5, 10, 15 μg/mL) of AHAVC- for 24 h. Cell proliferation and toxicity assay (cell counting kit-8, CCK-8) was used to detect the inhibition rates of the cells in different concentrations of AHVAC-. The migration ability of the cells was evaluated by wound-healing and Transwell assay. The apoptosis were observed by laser confocal microscopy with annexin V-mCherry/DAPI double staining, and the apoptosis rates were analyzed by flow cytometry with annexin V-FITC/PI double fluorescence staining. The protein level of Caspease-3 was determined by Western blot. RESULTS: Compared with normal control group, the results of AHVAC- concentration groups showed that with the increase of AHVAC- concentration, the proliferative activity of MN-28 cells decreased gradually (P<0.01), the cell migration ability decreased gradually (P<0.01), and the cell apoptosis rate increased (P<0.05). The expression of apoptosis-related protein Caspease-3 was up-regulated (P<0.01). CONCLUSION: AHVAC- inhibits proliferation and migration of gastric cancer MSN-28 cells and induces apoptosis.

Objective To screen the liver cell proteins interacting with the novel protein encoded by the 2.2 kb singly spliced variant of hepatitis B virus genome. Methods The splicing-specific gene TPss generated by the 2.2 kb singly spliced variant of HBV genome was amplified by PCR and cloned into the bait vector pGBKT7. After exclusion of self-activatian capacities of TPss protein, a two-hybrid library screening was performed using a pre-transformed human liver cDNA library to screen the liver cell proteins interacting with TPss. Mammalian two-hybrid assay was also done to further confirm the interactions between the bait and prey proteins in Huh7 and HepG2 hepatacytes. Results TPss gene with the size of 336 bp was successfully amplified and cloned, and the TPss protein expressed well in AHI09 yeast cells. Four liver cell proteins interacting with TPss, i. e. , cathepsin B, epoxide hydrolase 1, cathepsin D and fibrinogen gamma chain, were screened by yeast two-hybrid assay and further confirmed by mammalian two-hybrid assay. Conclusion TPss could interact with liver cell proteins.

Objective: To analyze CT and MRI manifestations of mucinous cystic neoplasms (MCN) and cyst-forming intraductal papillary neoplasms of the bile duct (IPNB). Methods: Clinicopathological and imaging data of 25 MCN patients (MCN group) and 16 cyst-forming IPNB patients (IPNB group, including 10 invasive IPNB patients and 6 noninvasive IPNB patients) confirmed pathologically were retrospectively analyzed. Results: There were significant differences of bile duct dilation, communication with lesions and bile ducts, mural nodules between MCN group and IPNB group (all P<0.001).No difference was found in lesions diameter, location, shapes, intracystic hemorrhage nor ADC value between the two groups (all P>0.05). Only lesion diameter was significantly different between invasive and noninvasive IPNB (P=0.032). Conclusion: CT and MRI features of MCN and cyst-forming IPNB are similar. Dilated bile ducts, communication between dilated bile ducts and mural nodules in lesions have high value in differential diagnosis of these two diseases.

Objective: To explore the diagnostic value of MRI in evaluating early aggressiveness of low grade central chondrosarcoma in long bones. Methods: Data of 11 patients with histopathologically proved WHO grade central chondrosarcoma in long bones were reviewed. All patients underwent conventional MRI within 2 weeks before operation. MRI findings of intracortical and intramedullary infiltration of chondrosarcoma were observed. Compared with surgical pathology, Kappa test was used to evaluate the consistency of MRI with pathology in diagnosis of early invasiveness of chondrosarcoma. The sensitivity, specificity and accuracy of MRI in diagnosis of intracortical and intramedullary infiltration of chondrosarcoma were calculated. Results: MRI showed 9 cases of chondrosarcoma with intracortical infiltration, among them 6 cases showed different degrees of scallop-like depression in the inner bone cortex, 3 cases had different forms of worm-like destruction in the inner bone cortex. Four cases of intramedullary infiltration showed blurred tumor edges and abnormal signals in the bone marrow around the tumors. MRI diagnosis of early aggressiveness of intramedullary and intracortical infiltration had good consistency with histopathological findings (Kappa=0.441, 0.621, both P<0.05). The sensitivity of MRI in diagnosis of intracortical and intramedullary infiltration was 90.00% (9/10) and 60.00% (3/5), specificity was 100% (1/1) and 83.33% (5/6), accuracy was 90.91% (10/11) and 72.73% (8/11), respectively. Conclusion: MRI plays an important role in detecting early aggressiveness of central chondrosarcoma in long bones.

Objective: To explore the value of DWI and dynamic contrast enhanced-MRI (DCE-MRI) quantitative parameters in evaluating the depth of myometrial invasion in patients with endometrial cancer. Methods Data of 45 patients with endometrioid adenocarcinoma confirmed by surgery and pathology were retrospectively analyzed. All patients underwent routine MRI, DWI and DCE-MRI 1-2 weeks before surgery. The patients were divided into no or superficial myometrial invasion group (n=25) and deep myometrial invasion group (n=20) according to the pathological results. The differences of ADC value and DCE-MRI quantitative parameters (Ktrans, Kep, Ve) were compared between the two groups. ROC curve was used to evaluate the diagnostic efficacy for depth of myometrial invasion in endometrial carcinoma. Results: Ktrans value of deep myometrial invasion group was higher than that of no or superficial myometrial invasion group (P=0.016). There was no significant difference in ADC value, Kep nor Ve between the two groups (all P>0.05). ROC curve analysis showed that the area under the curve of Ktrans was 0.735 (P=0.007). Taken Ktrans=0.355/min as the threshold, the sensitivity was 80.0%, and the specificity was 60.0% in diagnosis of depth of myometrial invasion in endometrial carcinoma. Conclusion: Ktrans may be useful for evaluating the depth of myometrial invasion of endometrial carcinoma.

Thyroid cancer is one of the most common malignant tumors. After standardized surgery, selective

OBJECTIVE: To investigate the role of long non-coding RNA ZEB1-AS1 in cerebral ischemia/reperfusion injury (CI/RI). METHODS: We detected the temporal changes of ZEB1-AS1 and HMGB1 expression using qPCR and Western blotting in SD rats following CI/RI induced by middle cerebral artery occlusion (MCAO). The rat models of CI/RI were subjected to injections of vectors for ZEB1-AS1 overexpression or knockdown into the lateral ventricle, and the changes in cognitive function, brain water content, blood-brain barrier integrity, and IL-1β and TNF-α levels in the cerebrospinal fluid (CSF) and serum were observed. Neuronal loss and cell apoptosis in the cortex of the rat models were detected by FJC and TUNEL methods, and HMGB1 and TLR-4 expressions were analyzed with Western blotting. We also examined the effects of ZEB1-AS1 knockdown on apoptosis and expressions of HMGB1 and TLR-4 in SH-SY5Y cells with oxygen-glucose deprivation/reoxygenation (OGD/R). RESULTS: In CI/RI rats, the expressions of ZEB1-AS1 and HMGB1 in the brain tissue increased progressively with the extension of reperfusion time, reaching the peak levels at 24 h followed by a gradual decline. ZEB1-AS1 overexpression significantly aggravated icognitive impairment and increased brain water content, albumin content in the CSF, and IL-1β and TNF-α levels in the CSF and serum in CI/RI rats (P < 0.05), while ZEB1-AS1 knockdown produced the opposite effects (P < 0.05 or 0.01). ZEB1-AS1 overexpression obviously increased the number of FJC-positive neurons in the cortex and enhanced the expressions of HMGB1 and TLR-4 in the rat models (P < 0.01); ZEB1-AS1 knockdown significantly reduced the number of FJC-positive neurons and lowered HMGB1 and TLR-4 expressions (P < 0.01). In SH-SY5Y cells with OGD/R, ZEB1-AS1 knockdown significantly suppressed cell apoptosis and lowered the expressions of HMGB1 and TLR-4 (P < 0.01). CONCLUSION ZEB1-AS1 overexpression aggravates CI/RI in rats through the HMGB1/TLR-4 signaling axis.
Animals ; Cell Line, Tumor ; Cell Proliferation/genetics* ; HMGB1 Protein/metabolism* ; Humans ; Infarction, Middle Cerebral Artery ; Neuroblastoma ; RNA, Long Noncoding/metabolism* ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; Toll-Like Receptor 4/metabolism* ; Tumor Necrosis Factor-alpha ; Water

Network pharmacology and liver fibrosis(LF) model in vitro were used to analyze the underly mechanism of anti-liver fibrosis effect that induced by Piperis Longi Fructus and its major active compounds. TCMSP and TCMIP were used to search for the chemical constituents of Piperis Longi Fructus, as well as the oral bioavailability(OB), drug-likeness(DL), intercellular permeability of intestinal epithelial cells(Caco-2) and Drug-likeness grading were set as limiting conditions. The related target genes of Piperis Longi Fructus were queried by TCMSP database, while related targets of LF were screened by GeneCards databases. Interaction network was constructed using Cytoscape 3.7.1. These above data were imported into STRING database for PPI network analysis. Enrichment of gene ontology(GO) and pathway analysis(KEGG) within Bioconductor database were utilized to note functions of related targets of Piperis Longi Fructus. Finally, the core targets and pathways were preliminarily verified by in vitro experiments. The effects of piperlongumine(PL), the major active component of Piperis Longi Fructus, on proliferation of rat liver stellate cells(HSC-T6) and expression of α smooth muscle actin(α-SMA) and collagen Ⅰ were investigated. The major factors TNF-α of tumor necrosis factor(TNF) pathway and NF-κB p65, IL-6 protein expressions of LF process were examined. A total of 12 active compounds such as PL were obtained by analyzing the bioavailability and drug-like properties, which inferred to 48 targets. The functional enrichment analysis of GO obtained 1 240 GO items, mainly involving in process of biology and molecular function. A total of 99 signaling pathways were enriched in the KEGG pathway enrichment analysis, including TNF signaling pathway, cGMP-PKG signaling pathway, calcium signaling pathways. CCK-8 assay showed that PL inhibited proliferation of HSC-T6 induced by transforming growth factor-β1(TGF-β1). Western blot analysis found that treated with PL suppressed the protein expressions of α-SMA, collagen Ⅰ, TNF-α and p65 in HSC-T6. Enzyme linked immunosorbent assay(ELISA) showed that PL inhibited the expressions of TNF-α and IL-6 in the cluture supertant of HSC-T6 cells. In conclusion, PL could play an anti-liver fibrosis role by regulating TNF/NF-κB signaling pathway. This study provided the mechanism basis of anti-LF effects induced by Piperis Longi Fructus and its major active compounds, which might help for the further study of the mechanism and key targets of Piperis Longi Fructus.
Animals ; Caco-2 Cells ; Hepatic Stellate Cells/metabolism* ; Humans ; Liver Cirrhosis/genetics* ; NF-kappa B/metabolism* ; Rats ; Signal Transduction

Objective To investigate the effects of the novel protein,TPds encoded by the 2.2 kb doubly spliced variant of hepatitis B virus(HBV)genome on the regulatory elements of HBV.Methods HBV promoters and enhancers were amplified by PCR and respectively cloned into the plasmid pGL3-basic to construct the recombinant firefly luciferase reporter vectors including pGL3-BCP(harboring basal core promoter),pGL3-CP1601(core promoter with enhancer Ⅱ),pGL3-XP(minimal X gene promoter),pGL3 XP1071(X gene promoter with enhancer Ⅰ),pGL3-SP1(promoter of large surface antigen)and pGL3-SP2 (promoter of middle surface antigen).The recombinant reporter plasmids were respectively co-transfected with either the pcDNA3.1/HisC-TPds(TPd8 expression plasmid)or pcDNA3.1/HisC(empty control)into Huh7 hepatocytes by FuGENE6 transfection reagent.Cells were lysed 48 h post transfection,the intracellulax luciferase activities were monitored and calculated by SPSS11.5 software.Results As compared with the empty control of pcDNA3.1/HisC.the intracellular luciferase activities were elevated when the Huh7 hepatocytes were co-transfected by pcDNA3.1/HisC-TPds with one of the recombinant reporter plasmids as pGL3-CP1601,pGL3-XP1071,pGL3-SP1,or pGL3-SP2,while no changes with pGL3-BCP or pGL3-XP.Conduslon TPds could transactivate the HBV regulatory elements a8 promoter SP1,SP2.and enhancer Ⅰ,Ⅱ,while no influences on basal core promoter and minimal X gene promoter.