Objective To observe the changes of peripheral blood interleukin-2(IL-2) ,interleukin-4(IL-4) ,interleukin-10(IL-10) ,tumor necrosis factor-alpha(TNF-α) and interferon gamma (IFN-γ) in chronic hepatitis B(CHB) patients with positive e-anti-gen(HBeAg) treated by Bushenqingtou Decoction .Methods 60 cases of CHB with positive HBeAg were randomly divided into the treatment group and the control group ,30 cases in each group .The CHB treatment group received Bushenqingtou Decoction with the treatment course of 48 week ,while the CHB control group did not received the medication therapy .In the beginning and at 48 weeks of treatment ,the blood in the two CHB groups were collected for detecting HBV DNA ,IL-2 ,IL-4 ,IL-10 ,TNF-αand IFN-γ. Results Compared with before treatments ,the levels of IL-2 ,TNF-αand IFN-γafter treatments in the CHB treatment group were obviously increased ,while the levels of HBV DNA ,IL-4 and IL-10 were remarkably decreased (P<0 .01);however ,there were no statistical differences of the various indexes after the treatments in the CHB control group (P>0 .05) .Conclusion Bushenqingtou Decoction could inhibit the replication of HBV DNA in the CHB patients with positive HBeAg and improve the body immune func-tion .

OBJECTIVE: To investigate the change in serum visfatin level after laparoscopic Roux-en-Y gastric bypass surgery in patients with Type 2 diabetes mellitus (T2DM) and to explore the relationship between visfatin insulin resistance and diabetes. METHODS: Thirty-three patients with Type 2 diabetes were studied before and after the gastric bypass surgery. The level of fasting serum visfatin was measured by enzyme-linked immunosorbent assay. Fasting plasma glucose (FPG), glycated hemoglobin (HbA1c) and fasting insulin (FINS) were measured before and after the gastric bypass surgery. RESULTS: Compared with before the operation, the indicators of HbA1c, FINS, and insulin resistance index (HOMA-IR) were decreased after the laparoscopic Roux-en-Y gastric bypass surgery. The body mass index (BMI) [(24.53 ± 0.62) kg/m² vs (26.71 ± 0.69) kg/m2] was decreased, with significant difference (P<0.001). The serum visfatin level [(9.79 ± 0.64) ng/mL] was significantly lower than before the operation [(38.24 ± 5.32) ng/mL], with significant difference (P<0.001). CONCLUSION Serum level of visfatin is decreased in T2DM patients who undergo gastric bypass surgry, reflecting an improvement in insulin resistance and diabetes.
Diabetes Mellitus, Type 2 ; blood ; surgery ; Female ; Gastric Bypass ; Humans ; Insulin Resistance ; Laparoscopy ; Male ; Middle Aged ; Nicotinamide Phosphoribosyltransferase ; blood ; Postoperative Period

Objective To investigate the diagnosis and differential diagnosis of dermatofibrosarcoma protuberans (DFSP). Methods Totally, 50 patients with DFSP visiting the Institute of Dermatology, Chinese Academy of Medical Sciences from 1998 to 2014 were enrolled. The clinical manifestations, histopathological and immunohistochemical features, treatment and prognosis of DFSP were retrospectively reviewed. Results The average age at onset of DFSP was (29.5 ± 15.9)years in the 50 patients, with a mean disease duration of 9.57 years. Skin lesions most frequently occurred on the trunk(n = 33, 66.0%), followed by the extremities, head and neck. DFSP was characterized by atrophic patches or plaques in 13 cases (26.0%), multiple nodules varying in size and arising on atrophic plaques or patches in 30 cases (60.0%), single or multiple nodules arising on normal skin in 7 cases (14.0%). Histologically, the tumor consisted of uniform infiltrative spindle cells arranged in a storiform or cartwheel pattern. In addition, the tumor cells expressed CD34 and vimentin. Twenty patients experienced recurrence at the primary site after resection of skin lesions with a recurrence rate of 43.5%. No distant metastasis or death occurred in these patients. Conclusions DFSP usually has various skin manifestations, is easily misdiagnosed, and can be confirmed based on histopathological and immunohistochemical findings. Local recurrence of DFSP is common, and may occur for many times after surgical excision, but lymphatic and distant metastases are rare.

Aim To explore the protective effects of LPPC ( procyanidins extracted from the litchi pericarp) on cardiomyocyte apoptosis in septic rats and its mech-anisms. Methods The rats were randomly divided in-to 5 groups, and were given orally the drug for two weeks continuously. The control group ( control) and sepsis model group ( LPS ) were given distilled water once a day. LPPC low, medium and high dose groups were given LPPC 50 , 100 , 200 mg · kg-1 · d-1 re-spectively which were prepared freshly every day. After the treatment, sepsis animal models were established. Except for the control group, other groups were injec-ted LPS (lipopolysacchride, 10mg·kg-1) intraperito-neally to induce acute sepsis model. 4hrs later, rat se-rum was collected, isoenzyme ( CK-MB ) , lactate de-hydrogenase ( LDH ) and activity of aspertate amin-otransferase ( AST/GOT) were detected. Then rat car-diac tissue was obtained and cardiac tissue malondial-dehyde ( MDA ) , total antioxidant capacity ( T-AOC ) and reduced glutathione ( GSH ) content were deter-mined. TUNEL staining was performed to analyze the apoptosis of myocardial cells. Cleaved caspase-3 and TNF alpha protein expressions were analyzed by West-ern blot. Results Compared with the control group ( control) , serum of sepsis model group rats CK-MB, LDH, AST/GOT and cardiac tissue MDA content were significantly increased (P<0. 01). At the same time, the activity of cardiac tissue T-AOC and GSH de-creased obviously ( P<0. 01 ) . The apoptotic myocar-dial cells increased significantly ( P<0. 01 ) , and the expression level of cleaved caspase-3 and TNF alpha decreased obviously ( P <0. 01 ) . LPPC pretreatment significantly decreased the serum CK-MB, LDH, AST/GOT and tissue MDA content, increased tissue T AOC and GSH activity, attenuated apoptosis of rat myocardi-al cells significantly, and decreased expression level of cleaved caspase-3 and TNF alpha. Conclusion LPPC pretreatment can significantly attenuate rat myocardial cell apoptosis induced by sepsis, and the underlying mechanisms may be related to its anti-oxidative effects.

Aim To study the mechanisms of the pro-tective effect of procyanidin B2 ( PCB2 ) on the myocar-dial cell apoptosis induced by lipopolysaccharide ( LPS) . Methods Using the primary culture rat myo-cardial cells, myocardial cell injury model was induced by LPS. PCB2 low, medium and high dose groups, were cultured with 6. 25 , 12. 5 , 25. 0 μmol · L-1 PCB2 respectively in DMEM medium for 24 h continu-ously. Myocardial cell survival rate was determined by MTT colorimetric method. Cardiacmyocyte NOX activi-ty was determined by lucigen chemiluminescence meth-od . Western blot analysis was used to detect myocardi-al NADPH oxidase p47phox expression. TUNEL method was used to detect apoptosis and flow cytometry was used to determine the content of myocardial cells ROS. Results Compared with control group, the cell dam-age induced by LPS group myocardial cell survival rate significantly decreased ( P <0. 01 ) , and myocardial cell NOX activity, p47phox expression, apoptotic cell number and ROS content were significantly increased (P<0. 01). PCB2 low, medium and high dose groups cell survival rates were significantly elevated, myocar-dial cell NOX activity and p47phox expression, apoptotic cell number and the ROS content decreased significant-ly in a dose-dependent manner ( P <0. 01 ) . Conclu-sion PCB2 protects myocardial cell apoptosis induced by LPS via inhibiting the expression of NADPH oxidase activation, p47phox expression and reactive oxygen spe-cies generation.

Objective:To investigate the feasibility of using shear wave dispersion (SWD) imaging in evaluating acute graft-versus-host disease (aGVHD) of the liver.Methods:A total of 42 Wistar rats were used as receptors and 10 Fischer 344 rats were used as donors for bone marrow transplantation to establish aGVHD models. Six rats were randomly selected every week for clinical observation and scoring. Then, ultrasonic SWD was performed to obtain shear wave speed (SWS) and shear wave dispersion slope (SWDS). Then, the histological characteristics were used as a reference standard, and the rats were divided into two groups: the group without aGVHD (nGVHD group) and the group with aGVHD. The differences in the clinical score and SWD values between the two groups were compared, the meaningful indexes were screened by binary Logistic regression analysis, and the joint prediction parameters were obtained. The ROC curve was plotted and the diagnostic efficiency was compared. The correlations between SWS, SWDS, clinical score and pathological grade were analyzed.Results:Clinical score, SWS, and SWDS in aGVHD group were higher than those in the nGVHD group (all P<0.05). The correlation between SWDS and pathological grade ( r=0.774, P<0.001) was higher than those between SWS, clinical score and pathological grade ( r=0.579, P=0.005; r=0.452, P=0.034). Logistic regression showed that SWDS was a significant indicator. In addition, the AUC values determined by SWDS and predictive parameters were (0.859, 0.886), which were significantly higher than the AUC of the clinical score (0.760, P<0.05). Conclusions:SWD imaging technology may become a promising method to evaluate hepatic aGVHD.

Objective To construct a recombinant lentiviral vector carrying the casitas B-lineage lymphoma (Cbl)-b short hairpin RNA (shRNA),and to evaluate its effect on the biological behavior of A375 melanoma cells in vitro.Methods Three specific shRNAs targeting Cbl-b gene and a negative control shRNA were designed and synthesized,and recombinant lentiviral vectors were constructed.A375 cells were divided into 5 groups to be transfected with 3 kinds of lentiviral vector expressing Cbl-b genespecific shRNAs (CBLB-shRNA-1 group,CBLB-shRNA-2 group and CBLB-shRNA-3 group),a lentiviral vector containing negative control shRNA (negative control group),and an empty vector (blank control group).Real-time fluorescence-based quantitative PCR and Western blot analysis were performed to determine the silencing efficiency at 72 hours after transfection.Cell counting kit-8 (CCK-8)assay was conducted to evaluate cellular proliferative activity at 24,48,72 and 96 hours after transfection,flow cytometry to detect cell apoptosis and cell cycle at 72 hours after transfection,and Transwell invasion assay to assess cellular invasive activity at 72 hours after transfection.Results Three recombinant lentiviral vectors containing Cbl-b shRNA were constructed successfully.As Western blot analysis revealed,the CBLB-shRNA-3 showed the highest silencing efficiency.CCK-8 assay indicated that the proliferative activity of A375 cells was significantly lower in the CBLB-shRNA-3 group than in the negative control group and blank control group at 72 and 96 hours after transfection(all P ＜ 0.01).Flow cytometry showed that the apoptosis rate of A375 cells was significantly higher in the CBLB-shRNA-3 group (22.73％ ± 6.58％) than in the negative control group (6.08％ ± 1.35％,P ＜ 0.01) and blank control group (6.34％ ± 1.07％,P ＜ 0.01).The CBLB-shRNA-3 group showed a significantly higher proportion of A375 cells at G1 phase,but a significantly lower proportion of A375 cells at S phase compared with the negative control group and blank control group(all P ＜ 0.01).Transwell assay showed that there were significant differences in the number of A375 cells crossing the artificial basement membrane (matrigel) at 72 hours after transfection among the negative control group,blank control group and CBLB-shRNA-3 group (76.60 ± 1.82,73.20 ± 3.83,19.60 ± 1.14,respectively;F =794.50,P ＜ 0.01).Conclusions A recombinant CBLB-shRNA-3-expressing lentiviral vector which can efficiently silence Cbl-b gene has been successfully constructed.It can inhibit the proliferation,cell cycle progression and invasive activity of A375 cells,but promote the apoptosis of A375 cells.

Objective To analyze differentially expressed proteins in A375 melanoma cells before and after short hairpin RNA (shRNA)-mediated Cbl-b gene silencing.Methods The label-free quantitative proteomics approach was performed to identify differentially expressed proteins between A375 cells transfected with lentiviral vectors containing Cbl-b shRNA (Cbl-b shRNA group) and those with control lentiviral vectors (control group).Then,the properties of differentially expressed proteins were analyzed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genome (KEGG) enrichment analysis.Western blot analysis was conducted to determine the expression of differential proteins (EphA2 and GSK3β) and phosphorylated protein kinase (p-AKT) after shRNA-mediated Cbl-b gene silencing.Statistical analysis was carried out by t test of two independent-samples for comparison of protein expression abundance between the two groups with SPSS 23.0 software.Additionally,the results of GO and KEGG enrichment analysis were analyzed by Fisher's exact test.Results A total of 3 449 proteins were identified and quantified,and 74 of them were differentially expressed between the Cbl-b shRNA group and control group.Compared with the control group,52 proteins were up-regulated and 22 were down-regulated in the Cbl-b shRNA group.GO enrichment analysis of differential proteins revealed that the top five significantly enriched biological processes were integrin-mediated cell adhesion,single-organism metabolic process,regulation of integrin-mediated cell adhesion,regulation of protein-targeting mitochondria and nucleic acid metabolic process.The top five significantly enriched molecular functions included DNA binding,2-iron,2-sulfur cluster binding,signaling receptor activity,cadherin binding and cell adhesion molecule binding.The top five significantly enriched cell components included nucleosome,DNA packaging complex,photoreceptor connecting cilium,DNA-protein complex and extracellular region part.KEGG enrichment analysis demonstrated that the top five significantly enriched melanoma-related signaling pathways were folate biosynthesis,axon guidance,extracellular matrix-receptor interaction,adherens junction and Wnt signaling pathways.As Western blot analysis revealed,the Cbl-b shRNA group showed lower protein expression of EphA2 (0.369),but higher protein expression of GSK3β (3.524) compared with the control group (1),which were consistent with the results of proteomics analysis.Additionally,the protein expression of p-AKT was down-regulated in Cbl-b shRNA group (0.453) compared with the control group (1).Conclusion Cbl-b may be involved in the occurrence of melanoma through a variety of biological pathways,and the EphA2/PI3K/AKT signaling pathway may be one important pathway.