Objective To investigate the expression and clinical significance of liver tissue hepatitis B virus covalently closed circular DNA ( HBV cccDNA) and three serum markers ( including serum HBV DNA, HBsAg, HBeAg) for chronic hepatitis B antiviral treatment.Methods 80 patients with chronic hepatitis B were collected, they were treated with telbivudine tablets,and the liver tissue HBVcccDNA and serum HBV DNA expression were detected by fluorescence quantitative PCR( FQ-PCR) .Serum HBsAg,HBeAg expressions were detected by enzyme linked immunosorbent assay ( ELISA) before treatment,12 weeks,28 weeks,44 weeks,52 weeks after treatment. Then,the correlation of liver tissue HBVcccDNA with serum markers was analyzed.Results Liver tissue HBVcccD-NA,serum DNA HBV,serum HBsAg and serum HBeAg were significantly decreased with the time of treatment,the differences were statistically significant(F=3.786,4.785,3.806,3.452,P=0.034,0.009,0.031,0.042),and the liver tissue HBVcccDNA, serum DNA HBV and serum HBeAg before treatment compared with after treatment had statistically significant differences(all P<0.05).And the differences of serum HBsAg in the treatment of 44 and 52 weeks compared with before treatment were statistically significant(all P<0.05).The results of correlation analy-sis showed that the expression of HBVcccDNA was positively correlated with serum HBV DNA and HBeAg ( r =0.674,0.672,P=0.015,0.036),and had no significant correlation with serum HBsAg expression(r=0.125,P=0.142 ) .Conclusion The expressions of serum HBV DNA and HBeAg could reflect HBVcccDNA expression of liver tissue,and the detection method is simple and non-invasive,which is worthy of recommendation in clinical.

Objective To investigate the effects of Gynostemma Pentaphyllum on serum markers of liver fibrosis in cholesterol-induced non-alcoholic fatty liver disease (NAFLD) in rabbits. Methods Forty adult male white rabbits were randomly divided into a normal control group, a model group, a treatment group and a simvastatin group, with 10 rabbits in each group. NAFLD was induced with a high-cholesterol diet. After modeling, the rabbits in the treatment group were intragastrically administrated with Gynostemma Pentaphyllum 5 mg/(kg?d), the simvastatin group with simvastatin 5 mg/(kg?d), and the model and normal control groups with the equal volume of distilled water for 9 weeks. The serum levels of total cholesterol (TC) and triglyceride (TG), the serum inflammatory maker C-reactive protein (CRP), the serum markers of liver fibrosis such as hyaluronic acid and laminin (LN), and the TG level in the liver tissue were detected. Results Compared with the normal control group, the serum levels of TC (60.50 ± 9.77 mg/L vs.1.30 ± 0.44 mg/L), TG (1.72 ± 0.61 mmol/L vs. 0.85 ± 0.39 mmol/L), CRP (256.79 ± 30.78 mg/L vs. 8.71 ± 1.41 mg/L), HA (798.8 ± 69.4 ng/ml vs. 121.2 ± 6.8 ng/ml),LN (964.8 ± 62.8 ng/ml vs. 142.4 ± 12.2 ng/ml) in the model group were increased significantly (all P<0.01). Compared with the model group, the serum levels of TC (36.44 ± 6.57 mmol/L vs. 60.50 ± 9.77 mmol/L), TG (1.31 ± 0.39 mmol/L vs. 1.72 ± 0.61 mmol/L), CRP (68.77 ± 10.78 mg/L vs. 256.79 ± 30.78 mg/L), HA (420.8 ± 60.2 ng/ml vs. 798.8 ± 69.4 ng/ml), LN (449.8 ± 56.6 ng/ml vs. 964.8 ± 62.8 ng/ml) and the TG level in the liver tissue (0.52 ± 0.10 mmol/L vs. 0.77 ± 0.08 mmol/L) in the treatment group were decreased significantly (all P<0.01). The TG level in the liver tissue in the treatment group was significantly lower than that in the simvastatin group (0.52 ± 0.10 mmol/L vs. 0.59 ± 0.09 mmol/L;P<0.05). There were no significant difference in the serum levels of TC, TG,CRP,HA and LN between the treatment group and the simvastatin group (all P>0.05). Conclusions Gynostemma Pentaphyllum may regulate lipid metabolism, alleviate inflammation, and decrease serum markers of liver fibrosis, and might protect against liver fibrosis in rabbits with NAFLD.

Objective To observe the influence ofgypenosides on hydrogen sulfide in liver tissue and plasma of rat with type 2 diabetes mellitus and nonalcoholic fatty liver disease.Methods 58 SPF male SD rats,with body mass 220～250 g,were randomly divided into a blank control group (group N,n=7),and a NAFLD and T2DM model group (Group M,n=51).Group N was fed with ordinary diet in the first four weeks,group M was fed with diets of high fat and sugar,injected with 40 mg/kg STZ overnight,and the same diets for the next four weeks.The rat model with T2DM and NAFLD was build.NAFLD and T2DM model group were divided into three groups:a high dose GPS group (JH,n=9) injected with 1 g/kg · d-1 GPS,a low dose GPS group (JL,n=9) injected with 0.5 g/kg · d-1 GPS,and a model group (M,n=9) injected with the same volume of water,and high fat diet at the same time.The treatment period was six weeks,and the experiment period was fourteen weeks.TG,TC,BS,and H2S in the plasma of rat were tested,and H2S in the liver tissue of the rat was tested.Results ①The changes of H2S in plasma:group JH [(4.30±0.43) μmol/L] and JL [(3.83 ±0.47) μmol/L] was lower than group M [(2.67 ± 0.41) μmol/L],there was a significant difference.②The changes of H2S in the liver tissue:group JH [(333.52±37.94) pmol/min/mg/protein] and JL [(275.81 ±36.07)pmol/min/mg/protein] was lower than group M [(237.8± 33.05) pmol/min/mg/protein],there was a significant difference.③BS levels:group JH(10.86±3.46)mmol/L,group JL (14.78±3.39)mmol/L,group M(18.84±4.24) mmol/L,group JH and JL was lower than group M,there was a significant difference (P＜0.01).④The plasma TG level:group N (0.96±0.09) mmol/L,group JH (2.82± 0.66) mmol/L,group JL (1.83± 0.56) mmol/L,group M (3.97 ± 0.64) mmol/L.group JH and JL was lower than group M,there was a significant difference (P＜0.01).Conclusion Gypenoside can reduce the blood sugar,triglycerides,and total cholesterol in rat with with type 2 diabetes mellitus and nonalcohol fatty liver disease.H2S concentrations in plasma and liver tissue of the rats with T2DM and NAFLD were increased by GPS,showing dose dependence.Gypenosides can also improve metabolism of blood glucose and lipid in rats with T2DM and NAFLD.

Objective To study the protein expressions of Fas-associated death domain protein (FADD) and caspase-8 in rats with intracerebral hemorrhage ,and the effects of erythropoietin tp reveal the mechanism of neu-m-protection by EPO. Methods 126 male SD rats were randomly divided into three groups: Sham-operated group, intracerebral hemorrhage group, and EPO group. Each group was divided into seven subgroups according to the differ-ent time points (3,6,12,24,48,72 h and 7 d). The model of intracerebral hemorrage was established in rats by in-tracerebral injection of autogenous blood. The protein expressions of FADD and caspas-8 in rats tissue around the hemorrhagic and the normal brain tissue were detected by immunohistochemistry. Results The protein expressions of FADD and caspase-8 were increased [(4.66±0.46 ) and ( 15.89±1.81)] at 3 h after intracerebral hemorrhage, and peaked at 48 h [ (35.88±4.24 ) and (45.04±3.99)], the expressions of FADD and caspas-8 in the region around hematoma in EPO group significantly decreased compared with model group[ (3.92±0.64) and (28.24±1.90), (13.32±2.01 ) and (35.08±2.82)] at 3 h and 48 h. Conclusion The protein expressions of FADD and easpase-8 are markedly increased after intracerebral hemorrhage. EPO can protect the neurons by signifi-cantly reducing the expressions of FADD and caspase-8.

Brucella is an intracellular pathogen that invades a host and settles in its immune cells; however, the mechanism of its intracellular survival is unclear. Modification of small ubiquitin-related modifier (SUMO) occurs in many cellular activities. E2 conjugating enzyme 9 (Ubc9) is the only reported ubiquitin-conjugating enzyme that links the SUMO molecule with a target protein. Brucella's intracellular survival mechanism has not been studied with respect to SUMO-related proteins and Ubc9. Therefore, to investigate the relationship between Brucella melitensis 16M and SUMO, we constructed plasmids and cells lines suitable for overexpression and knockdown of SUMO1 and Ubc9 genes. Brucella 16M activated SUMO1/Ubc9 expression in a time-dependent manner, and Brucella 16M intracellular survival was inhibited by SUMO1/Ubc9 overexpression and promoted by SUMO1/Ubc9 depletion. In macrophages, Brucella 16M-dependent apoptosis and immune factors were induced by SUMO1/Ubc9 overexpression and restricted by SUMO1/Ubc9 depletion. We noted no effect on the expressions of SUMO1 and Ubc9 in B. melitensis 16M lipopolysaccharide-prestimulated mouse RAW264.7 macrophages. Additionally, intracellular survival of the 16M△VirB2 mutant was lower than that of Brucella 16M (p < 0.05). VirB2 can affect expression levels of Ubc9, thereby increasing intracellular survival of Brucella in macrophages at the late stage of infection. Collectively, our results demonstrate that B. melitensis 16M may use the VirB IV secretion system of Brucella to interact with SUMO-related proteins during infection of host cells, which interferes with SUMO function and promotes pathogen survival in host cells.
Animals ; Apoptosis ; Brucella melitensis ; Brucella ; Immunologic Factors ; Macrophages ; Mice ; Plasmids

BACKGROUND:In recent years,the incidence of hyperuricemia caused by purine metabolism disorders has been increasing,which can induce inflammatory responses and lead to renal injury. OBJECTIVE:To explore the role and mechanism of solute carrier family 2 member 12(SLC2A12)in hyperuricemia-related renal injury. METHODS:Renal tubular cells(HK2 cells)were divided into five groups:HK2 group,HK2+uric acid group,HK2+uric acid+NC group,HK2+uric acid+siSLC2A12 group,and HK2+uric acid+siSLC2A12+MK-2206 group.HK2 cells were treated with uric acid and transfected with siRNA SLC2A12,followed by MK-2206 treatment to inhibit AKT expression.Cell proliferation was detected by CCK-8 assay.Apoptosis was detected by TUNEL assay.qRT-PCR and western blot assay were used to detect fibrogenic factors as well as activation of the AKT/FOXO3a pathway.The concentrations of inflammatory cytokines were measured by enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION:(1)Uric acid treatment inhibited cell proliferation and promoted cell apoptosis in the HK2+uric acid group compared with the HK2 group.The proliferative ability of cells in the HK2+uric acid+siSLC2A12 group was further decreased and apoptotic cells were further increased compared with the HK2 group.Compared with the HK2+uric acid+siSLC2A12 group,the HK2+uric acid+siSLC2A12+MK-2206 group showed an increase in cell proliferation and a decrease in apoptotic cells.(2)Compared with the HK2 group,the connective tissue growth factor(CTGF),α-smooth muscle actin(α-SMA)and transforming growth factor beta(TGF-β)expressions increased in the HK2+uric acid group;CTGF,α-SMA and TGF-β expression further increased in the HK2+uric acid+siSLC2A12 group.Compared with the HK2+uric acid+siSLC2A12 group,the CTGF,α-SMA and TGF-β expressions decreased.(3)Compared with the HK2 group,the expression of p-AKT,FOXO3a,and p-FOXO3a elevated in the HK2+uric acid group;the expression of p-AKT further increased,while the expression of FOXO3a and p-FOXO3a decreased in the HK2+uric acid+siSLC2A12 group.Compared with the HK2+uric acid+siSLC2A12 group,p-AKT expression decreased;FOXO3a and p-FOXO3a expression increased in the HK2+uric acid+siSLC2A12+MK-2206 group.(4)Compared with the HK2 group,interleukin-6,interleukin-1 β,and tumor necrosis factor α levels increased in the HK2+uric acid group;interleukin-6,interleukin-1 β,and tumor necrosis factor α levels further increased in the HK2+uric acid+siSLC2A12 group.Compared with the HK2+uric acid+siSLC2A12 group,interleukin-6,interleukin-1 β,and tumor necrosis factor α levels diminished in the HK2+uric acid+siSLC2A12+MK-2206 group.(5)These findings indicate that SLC2A12 may protect against hyperuricemia-induced renal injury by counteracting uric acid-induced tubular fibrosis and inflammation through activation of the FOXO3a pathway.

The rapidly developing resistance of cancers to chemotherapy agents and the severe cytotoxicity of such agents to normal cells are major stumbling blocks in current cancer treatments. Most current chemotherapy agents have significant cytotoxicity, which leads to devastating adverse effects and results in a substandard quality of life, including increased daily morbidity and premature mortality. The death receptor of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) can sidestep p53-dependent pathways to induce tumor cell apoptosis without damaging most normal cells. However, various cancer cells can develop resistance to TRAIL-induced apoptosis via different pathways. Therefore, it is critical to find an efficient TRAIL sensitizer to reverse the resistance of tumor cells to TRAIL, and to reinforce TRAIL's ability to induce tumor cell apoptosis. In recent years, traditional Chinese medicines and their active ingredients have shown great potential to trigger apoptotic cell death in TRAIL-resistant cancer cell lines. This review aims to collate information about Chinese medicines that can effectively reverse the resistance of tumor cells to TRAIL and enhance TRAIL's ability to induce apoptosis. We explore the therapeutic potential of TRAIL and provide new ideas for the development of TRAIL therapy and the generation of new anti-cancer drugs for human cancer treatment. This study involved an extensive review of studies obtained from literature searches of electronic databases such as Google Scholar and PubMed. "TRAIL sensitize" and "Chinese medicine" were the search keywords. We then isolated newly published studies on the mechanisms of TRAIL-induced apoptosis. The name of each plant was validated using certified databases such as The Plant List. This study indicates that TRAIL can be combined with different Chinese medicine components through intrinsic or extrinsic pathways to promote cancer cell apoptosis. It also demonstrates that the active ingredients of traditional Chinese medicines enhance the sensitivity of cancer cells to TRAIL-mediated apoptosis. This provides useful information regarding traditional Chinese medicine treatment, the development of TRAIL-based therapies, and the treatment of cancer.

BACKGROUNDNon-small cell lung cancer (NSCLC) is one of the most common malignant tumors. Despite the advances in therapy over the years, its mortality remains high. The aim of this study was to evaluate the expression of small ubiquitin-like modifier (SUMO) proteases 1 (SENP1) in NSCLC tissues and its role in the regulation of vascular endothelial growth factor (VEGF) expression. We also investigated the association between the expression level of SENP1 and the clinicopathological features and survival of the patients.

METHODSA SENP1 small interfering RNA (siRNA) was constructed and transfected into the NSCLC cells. VEGF gene expression was analyzed by real-time polymerase chain reaction (RT-PCR). Immunohistochemistry staining was used to assess the expression of SENP1 in 100 NSCLC patients and its association with the clinicopathological features and survival was analyzed.

RESULTSVEGF expression was significantly higher in NSCLC tissues than in normal lung tissues. Inhibition of SENP1 by siRNA was associated with decreased VEGF expression. SENP1 was over-expressed in 55 of the 100 NSCLC samples (55%) and was associated with a moderate and low histological tumor grade (3.6%, 38.2%, and 58.2% in high, moderate and low differentiated tumors, respectively, P = 0.046), higher T stage (10.9% in T1, and 89.1% in T2 and T3 tumor samples, P < 0.001) and TNM stage (10.9% in stage I, and 89.1% in stages II and III tumor samples, P < 0.001). The rate of lymph node metastasis was significantly higher in the SENP1 over-expression group (76.4%) than that in the SENP1 low expression group (33.3%, P < 0.001). Sixty three patients received postoperative chemotherapy, including 34 with SENP1 over-expression and 29 with SENP1 low expression. Among the 34 patients with SENP1 over-expression, 22 (64.7%) patients developed recurrence or metastasis, significantly higher than those in the low expression group 27.6% (8/29) (P = 0.005). Multivariate Cox regression analysis showed that lymph node metastasis (P = 0.015), TNM stage (P = 0.001), and SENP1 expression level (P = 0.002) were independent prognostic factors for the survival of NSCLC patients.

CONCLUSIONSSENP1 may be a promising predictor of survival, a predictive factor of chemo-sensitivity for NSCLC patients, and potentially a desirable drug target for lung carcinoma target therapy.

Antineoplastic Agents ; therapeutic use ; Blotting, Western ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; genetics ; metabolism ; Cell Line, Tumor ; Cysteine Endopeptidases ; Endopeptidases ; genetics ; metabolism ; Female ; Humans ; Immunohistochemistry ; In Vitro Techniques ; Lung Neoplasms ; drug therapy ; genetics ; metabolism ; Male ; Reverse Transcriptase Polymerase Chain Reaction