Objective To investigate the distribution of arterial stenosis in patients with ischemic cerebrovas-cular disease and its risk factors .Methods 224 patients with ischemic cerebrovascular disease were divided into four groups according to DSA results .Patients showed no stenosis or mild stenosis were selected as control group ( 43 ca-ses),the other patients were divided into pure extracranial stenosis group (41 cases),simple intracranial stenosis group (93 cases) and extracranial stenosis group (47 cases).The results of laboratory test were analyzed .The ques-tionnaire was designed and the indicators including name ,age,long-term smoking,drinking,hypertension and diabetes were collected .Results Of all the subjects , there were 181 cases with artery stenosis .Single factor results showed that gender,age,long-term smoking,long-term drinking,hypertension,diabetes,high homocysteine,high level of lyso-phosphatidic acid were the independent risk factors of cerebral artery stenosis (χ2/t =8.744, 5.562, 10.736, 11.032,9.812,10.002,9.083,2.576,all P<0.05).Multivariate analysis showed that high homocysteine ,long-term smoking and drinking were the risk factors of simple intracranial artery stenosis .High homocysteine and high fibrino-gen were risk factors of simple extracranial stenosis .Age,hypertension,long-term smoking and drinking ,high homocys-teine and fibrinogen were risk factors of intracranial and extracranial stenosis .Conclusion In patients with ischemic cerebrovascular diseases ,the simple intracranial artery stenosis is most common .The incidence of cerebral artery ste-nosis has age characteristic ,with the increase of age ,the incidence rate of intracranial and extracranial artery stenosis is rising.High homocysteine,long-term smoking and drinking,hypertension,diabetes,high lysophosphatidic acid are independent risk factors of cerebral artery stenosis .

BACKGROUND:Angiotensin II receptor antagonists have been found to exerct a stronger protective effect on bone than angiotensin converting enzyme inhibitors. OBJECTIVE:To investigate the effect of different concentrations of irbesartan (angiotensin II receptor antagonist) on the differentiation and mineralization of mouse preosteoblasts. METHODS:Mouse preosteoblast cel lines MC3T3-E1 in logarithmic phase were selected and cultured in the osteogenic induction medium containing 0 (control group), 0.001, 0.01, 0.1 mmol/L irbesartan, respectively. Ten days later, the cel differentiation was observed by alkaline phosphatase staining. The mineralization was observed by alizarin red staining after 21 days of culture. mRNA expressions of osteocalcin, alkaline phosphatase and Runt-associated transcription factor 2 in osteoblasts were detected by real-time PCR at 1, 4, 7, 14 and 21 days of culture. RESULTS AND CONCLUSION:The activity of alkaline phosphatase in al the irbesartan groups (0, 0.001, 0.01, 0.1) was higher than that in the control group (P<0.05), which was the most obvious in 0.01 mmol/L. The number and area of calcium nodules in each irbesartan group were significantly higher than those in the control group (P<0.05), especial y in 0.01 mmol/L. Compared with the control group, 0.01 mmol/L irbesartan significantly upregulated the mRNA expressions of osteocalcin, alkaline phosphatase and Runt-associated transcription factor 2 (P<0.05). These results suggest that 0.01 mmol/L irbesartan significantly promotes the differentiation and mineralization of osteoblasts.

Objective:To investigate the mechanisms of TWS119 induced CCR5 expression in hunman γδT cells. Methods:After treatment with various concentrations of TWS119 for 48h, the expression of CCR5 in γδT cells were detected by flow cytometry. The p-STAT3 and GAPDH expression were examined by Western blot analysis. Results: TWS119 could upregulate the expression of CCR5 in dose dependent manner. Western blot analysis revealed that TWS119 inhibit phosphorylation of STAT3,but had no significant impact on GAPDH. In addition, pretreatment of γδT cells with 0. 5 μmol/L STAT3 specific phosphorylation inhibitor Stattic could upregulate the expression of CCR5 and enhance the TWS119 induced CCR5 expression. Conclusion: TWS119 could upregulate CCR5 expression of γδT cells by inhibiting STAT3 phosphorylation in vitro.

Objective:To investigate the anti-tumor effect of Hyperoside.Methods: Human γδT cells were amplified by isopentenyl pyrophosphate from peripheral blood cells.The proliferation capacity of γδT cells was measured with CCK-8 assay after treated with different concentrations of Hyperoside.Cytotoxicity of γδT cells was detected with LDH assay , and the expression of granzyme,perforin CD107a and IFN-γonγδT cells were measured by flow cytometry before and after treatment.Results: Hyperoside could significantly stimulate the proliferation of γδT cells at the concentration of 3.13-12.5 μg/ml.Cytotoxicity and expression of granzyme,perforin and IFN-γofγδT cells were increased after treatment.Conclusion:Hyperoside could enhance cytotoxicity of humanγδT cells through up-regulation of granzyme ,perforin CD107 a and IFN-γexpression.

Objective To discuss the clinical value of magnetic resonance diffusion-weighted imaging (MR-DWI) in distinguishing tumor remnants from tumor necrosis of pancreatic carcinoma after cryoablation treatment.Methods Conventional MRI T1WI,T2WI scan,DWI sequence and dynamic enhanced MRI scan were performed in 26 patients with pancreatic carcinoma who were received cryoablation treatment.The changes in MRI signals after cryoablation treatment were recorded.The apparent diffusion coefficient (ADC) values of the normal pancreas,preoperative tumor tissue,postoperative remnants and necrosis tissue were calculated,and the results were compared.The correlation between the ADC values and the size of the tumor was evaluated,and the differences in ADC values among the tumors that had different diameter,location and staging were statistically analyzed.Results Of the 26 patients,complete necrosis of tumor was obtained in 16.The necrotic tumor tissue displayed low-signal on T1WI,high-signal on T2WI and low-signal on DWI,with no enhancement on dynamic enhanced imaging.Active residual tumor tissue was detected in 9 patients,among them the residual tumor diameter ＞5 cm was seen in 7 patients;the residual rate was 34.6％.ADC values of the following tissue,from low to high in order,were preoperative pancreatic tumor tissue (1.022± 0.126)x10-3 mm2/s,postoperative residual tumor tissue (1.130±0.155)x10-3 mm2/s,normal pancreatic tissue (1.924±-0.124)×10-3 mm2/s and postoperative necrosis tissue (2.312-±0.214)×10-3 mm2/s.No statistically significant difference in ADC values existed between preoperative pancreatic tumor tissue and postoperative residual tumor tissue (P=0.452),while statistically significant difference in ADC values existed between normal pancreatic tissue and postoperative necrosis tissue (P＜0.001).The ADC values of pancreatic tumor tissue bore a negative correlation with the tumor size (R=-0.43,P=0.027 2),while the ADC values lacked the relationship to the tumor location as well as to the tumor staging (P=0.738 8 and P=0.089 5 respectively).Conclusion MR-DWI can effectively distinguish the residual tumor tissue from the necrotic tumor tissue of pancreatic carcinoma after cryoablation treatment,which provides reliable basis for further clinical diagnosis and treatment.

Objective:To investigate the effect of glycogen synthase kinase-3β inhibitor TWS119 on hunman γδT cells growth and phenotypic characteristics. Methods:Using γδT medium to cultivate human peripheral blood γδT cells in vitro. After co-cultured with different concentrations of glycogen synthase kinase-3β( GSK-3β) inhibitor 4, 6-disubstituted pyrrolopyrimidine ( TWS119 ) for indicated time,growth curve and Wnt/β-catenin activation of in each group were determined by CCK-8 and Western blot assays. The CD62L and CD45RA expression theγδT cells were detected using flow cytometry. Results:Wnt/β-catenin pathway ofγδT cells could activate by TWS119. In the first group,TWS119 could upregulate the expression of CD62L and CD45RA in dose dependent manner while inhibit the growth and ratio of γδT cells. In the second group,TWS119 could promote the growth and ratio of γδT cells with the increase in concentration from 0 μmol/L to 4. 0 μmol/L and decreased thereafter. Besides,TWS119 could promote the expression of CD62L in a dose-dependent and had no effect on the CD45RA. Conclusion: Human γδT cells isolated from peripheral blood treated with TWS119 gave rise to two subsets of CD45RA+CD62L+γδT cells and CD62L+γδT cells.

Objective:To investigate the effect of 2-deoxy-D-glucose (2-DG) on hunmanγδT cells on the expression of CCR5 and killing function in vitro.Methods:UsingγδT medium to cultivate peripheral blood mononuclear cell( PBMCs) in vitro.After co-cultured with various concentrations of 2-DG for 48 h,the expression of CCR5 and killing activities of γδT cells for each group were detected by flow cytometry and CCK-8 methods.Results: 2-DG could not promote the growth of γδT cells with the increase in concentration from 0 μmol/L to 1.0 μmol/L and decreased thereafter.The certain concentration ( 0-2.0 μmol/L ) of 2-DG could upregulate the expression of CCR5 in dose dependent manner.Besides,at 0.5μmol/L and 1.0μmol/L of 2-DG could increase the ex-pression of CD107a and perforin and have no effect on the granzyme B.Conclusion: Human γδT cells isolated from peripheral blood treated with 2-DG could promote the expression of CCR5 and increase the killing activities at certain concentration in vitro.

Objective To investigate the effect of hyperoside on proliferation and killing activity of NK cells against pancreatic cancer PANC1 cells in vitro,and explore its potential mechanism.Methods Peripheral blood mononuclear cells of healthy donors were isolated,NK cells were induced with medium contained with IL-2 and different concentrations of hyperoside (0.3,1.6,8,40 and 200 μg/ml) for 12 days.Cell viability was observed by trypan blue staining.Phenotype and perforin,granzyme B expression of NK cells were detected by flow cytometry.Killing activity of NK cells against PANC1 cells were analyzed with lactate dehydrogenase (LDH) releasing method.Results The proportion of NK cells in control group and experimental group treated with different concentration of hyperoside both reached about 80％,respectively.The proliferation of CDs-CD56 + NK cells treated by hyperoside at 0.3,1.6 and 8 μg/ml was (93.76 ±8.77),(106.67 ± 12.35) and (118.50 ± 11.51) times,respectively,which were significantly higher than (73.70 ± 9.43) times of the control group.The expressions of perforin in NK cells treated with hyperoside at 1.6,8 and 40 μg/ml were significantly higher than those of the control group [(82.34 ± 2.90) ％,(89.15 ±3.54) ％,(81.78 ± 2.81)％ vs (72.93 ± 2.06)％].The expressions of granzyme B in NK cells treated with hyperoside at 1.6 and 8 μg/ml were significantly higher than those of the control group [(87.30 ± 1.70) ％,(92.16 ±3.05)％ vs (82.35 ±2.73)％].The killing activity of NK cells against PANC1 cells treated by hyperoside at 1.6 and 8 μg/ml was significantly higher than those of the control group [(63.18 ± 3.77)％,(65.34 ± 4.97) ％ vs (52.16 ± 5.48) ％].The differences were statistically significant (all P ＜ 0.05).Conclusions Hyperoside could promote the proliferation of NK cells at certain concentrations and maybe enhance the killing effect against pancreatic cancer PANC1 cells through up-regulating the expression of perforin and granzyme B in NK cells.

Objective To explore the safety and short?term efficacy of irreversible electroporation (IRE)ablation which is a novel ablation technology in unresectable hepatic neoplasms. Methods Patients with pathologically diagnosed as liver cancer or liver metastases were prospectively enrolled. The patients were not suitable for surgery with PS score ≤ 2. Exclusion criteria included who was not tolerate general anesthesia, severe liver and kidney dysfunction, and with cardiac pacemaker. A total of 16 patients were included in this study. There was 12 males and 4 females, aged 40 to 86 years with mean age (60 ± 10)y. Ultrasound and CT guided percutaneous IRE ablation was performed. Perioperative hemodynamic changes were reviewed. Liver and kindey function before and 7 d after ablation was compare by t test. The adverse reactions within 30 d after ablation treatment were recorded. CT and MR scans within 1 month were performed and the 30 d curative effect was evaluated by the modified RECIST criteria. Results All patients received IRE treatment successfully, and some patients experienced adverse reactions within 30 days after ablation, including abdominal pain in 7 cases, peritoneal effusion in 5 cases, hydrothorax in 4 cases, fever in 3 cases, cough, nausea and vomiting in 2 cases, biliary tract infection and thrombocytopenia in 1 case. After symptomatic treatment, these symptoms were improved. Severe complications, such as massive haemorrhage and bile leakage didn't occur. At 30 days after ablation, the curative effects were evaluated. Complete response (CR) was achieved in 1 patient , partial response (PR) was achieved in 12 patients, stable disease (SD) was in 2 patients , and progressive disease(PD) was 1 patients . The tumor relief rate (complete response+partial response) was 81.3%. Conclusions IRE ablation in the treatment of unresectable hepatic malignant tumor could have many advantages, including high safety, mild adverse reactions, and short?term efficacy. However, its long?term effect still need further observation.

Penicillium decumbens T. is an important filamentous fungus for the production of cellulases to effectively degrade lignocellulose for second generation biofuel production. In order to enhance the capability of Penicillium decumbens to produce cellulases, we constructed a creB (a deubiquitinating enzyme encoding gene) deletion cassette, and generated a creB knockout strain with homologous double crossover recombination. This mutation resulted in a detectable decrease of carbon catabolite repression (CCR) effect. The filter paper activity, endoglucanase activity, xylanase activity and exoglucanase activity of the deltacreB strain increased by 1.8, 1.71, 2.06 and 2.04 fold, respectively, when comparing with the parent strain Ku-39. A 2.68 fold increase of extracellular protein concentration was also observed. These results suggest that the deletion of creB results in CCR derepression. These data also suggest that CREB influences cellulase production of Penicillium decumbens. In generation, this study provides information that can be helpful for constructing cellulase hyper-producing strain.
Cellulase ; biosynthesis ; Endopeptidases ; genetics ; metabolism ; Gene Knockout Techniques ; Lignin ; metabolism ; Mutant Proteins ; metabolism ; Penicillium ; enzymology ; genetics ; Recombination, Genetic ; Ubiquitinated Proteins ; genetics ; Ubiquitination