Objective To explore the role of eNOS and NF-?B in the cultured endothelial cells apoptosis induced by TNF-? and Ang II.Methods The cultured endothelial cells were treated with TNF-? and Ang II in absence and presence of PDTC(an inhibitor of NF-?B);the mRNA of eNOS was measured by RT-PCR,the protein of eNOS and I?B? were assessed by Western-blot,the activity of NF-?B was evaluated by EMSA,and the apoptosis of cells was detected with Tunel.Results The mRNA and protein of eNOS were significantly decreased in the endothelial cells induced by TNF-? and AngII(P

Objective To investigate the relation between activator protein-1(AP-1)and coronary atheroselerotic changes and the potential role of AP-1 in the stabilization of atherosclerotic plaques in patients with coronary heart disease(CHD).Method 142 patients were included in this study and divided into CHD group(107)and control group(35)according to coronary angiography(CAG).The CHD group was further divided into a stable angina pectoris(SAP)group(32)and all acute coronary syndrome(ACS)group (75)according to the clinical manifestations.In addition,the CHD group was divided into A type group,B type group and C type group according to the standard of ACC/AHA coronary change in 1988.Meanwhile,the CHD group was further divided into light stenosis group,moderate stenosis group and severe stenosis group according to the degree of coronary lesion.The lysate of cells was obtained through lysis of the leucocyts from peripheral blood with cell lysis buffer.The amount of Phospho.c-Jun in lysate was measured with enzyme-linked immunosorbent assay(ELISA).The results were demonstrated with absorbance,which reflects the amount of AP-1.Results The main coronary changes in the SAP group were A type(68.7%)and the changes were mainly of light degree(53.1%);the main coronary changes in the ACS group were B type(52.0%)or C type(37.3%)and the changes were mainly of heavy degree(66.7%).The absorbance of Phospho-c-Jun in CHD group was significantly higher than that in the control subjects (1.43±0.33 vs 0.71±0.13,P＜0.001).The absorbance of Phospho-c-Jun in the ACS group was significantly higher than that in the SAP group(1.56±0.28 vs 1.14±0.25,P＜0.001).The absorbance of Phospho-c-Jun increased gradually from A type group to C type group(1.18±0.27 vs 1.42±0.26 vs 1.71±0.27,P＜0.001)and from light stenosis group to severe stenosis group(1.09±0.20 vs 1.37±0.26 ys 1.60±0.29,P＜0.001).Conclusion There is a significant relationship between AP-1 and coronary atherosclerotic changes.AP-1 may be a factor that can predict coronary arteriosclerotic progression and stability of the plaque.

Objective To investigate the relationship between the plasma adiponectin concentration and coronary arteriosclerosis change in patient with coronary heart disease(CHD).Method 142 patients were divided into CHD group and control group according to the Coronary Angiography(CAG).CHD group were further divided into stable angina pectoris(SAP)subgroup and acute coronary syndrome (ACS)subgroup according to the clinical property.According to the type of coronary change,CHD group wag divided into A type group, B type group and C type group,meanwhile according to the degree of coronary lesion,CHD group was divided into light stenosis group, moderate stenosis group and severe stenosis group.The plasma adiponectin concentration was measured by ELISA.Results The plasma adiponectin concentration in CHD group was significant lower than that in control group.The plasma adiponectin concentration in ACS subgroup was significant lower than that in SAP subgroup.The plasma adiponectin concentration decreased gradually from A type group to C type group and from light stenosis group to severe stenosis group(P＜0.001).Conclusions Adiponectin is a negative regulatory factor of coronary atherosclerosis,and Hypoadiponectin may be used to predict the change of coronary arteriosclerosis and the stability of plaque.

Objective To observe the efficacy and adverse reaction of ozone therapy combined with gemcitabine and cisplatin (GP) regimen in patients with advanced non-small cell lung cancer.Methods Fifty-five patients with advanced non-small cell lung cancer were enrolled and allocated to treatment group (28 cases) and control group (27 cases).The patients in treatment group received ozone therapy combined with GP regimen,and the patients in control group received GP regimen only.The efficacy,quality of life,adverse reaction and cellular immune function after treatment was compared between two groups.Results There was no significant difference in the efficacy between two groups (P ＞ 0.05).The quality of life after treatment in treatment group was better than that in control group (P ＜ 0.05).The liver function damage in treatment group was lower than that in control group (P ＜ 0.05).The cellular immune function in treatment group was stronger than that in control group (P ＜ 0.05).Conclusion Ozone therapy combined with GP regimen can effectively alleviate adverse induced by GP regimen chemotherapy and significantly improve the quality of life in patients with advanced non-small cell lung cancer.

AIM:The purpose of the present study was to investigate the effect of interleukin-10(IL-10)on the proliferation and calcineurin(CaN)activity in cultured cardiac fibroblasts(CFs)induced by arginine vasopressin(AVP).METHODS:The CFs of left ventricle in neonatal Sprague-Dawley rats were isolated and cultured by trypsin digestion and selective plating technique.Then the proliferation rates of cells were determined by using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay(A490 value).Cell cycle distribution was determined with flowcytometry technique.The CaN activity was measured by ultra-violet spectrophotography.RESULTS:(1)MTT colorimetry showed that 10-7 mol/L AVP significantly increased A490 value of CFs in comparison with control group(P

Objective To investigate the cellular and humoral immune responses and protective effect induced by co-immunization with two multi-epitope combinant antigens. Methods Mice were co-im-munized with the muhi-epitope HCV-T and HCV-E1 antigens three times. Sera antibodies IgG, IgG1 and IgG2a were tested by ELISA. Spleens from BALB/c mice immunized were removed 10 days after the last im-munization. CTL activity was assessed using LDH cytotoxicity assay kit. IFN-γ- and IL-4-secreting cells were quantified using ELISPOT kit. Two weeks after the final immunization, the mice were challenged sub-cutaneously(s, c. ) at the back with 106 SP2/0-NS3 cells, and protective effect was observed. For therapy, 106 SP2/0-NS3 cells were implanted into the back of BALB/c mice. Seven days later, mice were immuniza-tion three times. Therapy effect was observed. Results Co-immunization with HCV-T and HCV-E1 induced high tiers of HCV-El-specific IgG, IgG1 and IgG2a antibodies, and high level of CTL activity. Synergistic effect in frequencies of both specific IFN-γ-secreting cells and IL-4-secreting cells was observed in mice co-immunized. Prophylactic as well as therapeutic administration of mT + mE1 in mice led to protecting mice against SP2/0-NS3 cells. These results suggested that mT + mE1 was potential as a prophylactic as well as therapeutic HCV vaccine. Conclusion Co-immunization with HCV-T + HCV-EI induced protective humor-al and cellular immune response. HCV-T + HCV-E1 was potential as a recombinant HCV vaccine.

We constructed different N-terminal truncated variants based on Bacillus acidopullulyticus pullulanase 3D structure (PDB code 2WAN), and studied the effects of truncated mutation on soluble expression, enzymatic properties, and application in saccharification. Upon expression, the variants of X45 domain deletion existed as inclusion bodies, whereas deletion of CBM41 domain had an effective effect on soluble expression level. The variants that lack of CBM41 (M1), lack of X25 (M3), and lack both of CBM41 and X25 (M5) had the same optimal pH (5.0) and optimal temperature (60 degrees C) with the wild-type pullulanase (WT). The K(m) of M1 and M5 were 1.42 mg/mL and 1.85 mg/mL, respectively, 2.4- and 3.1-fold higher than that of the WT. k(cat)/K(m) value of M5 was 40% lower than that of the WT. Substrate specificity results show that the enzymes exhibited greater activity with the low-molecular-weight dextrin than with high-molecular-weight soluble starch. When pullulanases were added to the saccharification reaction system, the dextrose equivalent of the WT, M1, M3, and M5 were 93.6%, 94.7%, 94.5%, and93.1%, respectively. These results indicate that the deletion of CBM41 domain and/or X25 domain did not affect the practical application in starch saccharification process. Furthermore, low-molecular-weight variants facilitate the heterologous expression. Truncated variants may be more suitable for industrial production than the WT.
Bacillus ; enzymology ; Glycoside Hydrolases ; metabolism ; Molecular Weight ; Protein Conformation ; Sequence Deletion ; Substrate Specificity ; Temperature

Objective To explore the effect of preoperative administration of glycopyrronium bromide on postoperative cognitive dysfunction(POCD)in elderly patients undergoing laparoscopic inguinal hernia repair surgery.Methods 74 elderly patients underwent laparoscopic inguinal hernia repair surgery under general anesthesia with a laryngeal mask were randomly divided into control group(group C,25 cases),low-dose glycopyrronium bromide group(0.2 mg,group L,24 cases),medium-dose glycopyrronium bromide group(0.4 mg,group M,25 cases).Blood was collected from patients 1 d before surgery and 3 d after surgery,and serum levels of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),brain function related factors[5-hydroxytryptamine(5-HT),brain-derived neurotrophic factor(BDMF)]were measured by enzyme linked immunosorbent assay(ELISA).The Montreal Cognitive Assessment Scale(MoCA)was used to assess the cognitive function of the patients.Airway secretion scores,the incidence of arrhythmia,dry mouth and dysuria were recorded.Results There was no significant difference in the incidence of POCD among the three groups of patients(P＞0.05);At 3 d postoperatively,5-HT and BDNF levels in group L were higher than those in group C,the BDNF level in group L was higher than that in group M,there were statistically significant(P＜0.05);At 3 days after surgery,the levels of serum IL-1β,IL-6 and TNF-α in group C were significantly higher than those at 1 day before surgery(P＜0.05),the levels of serum IL-6 and TNF-α in group M were significantly higher than those at 1 day before surgery(P＜0.05),there were no significant differences in the levels of serum IL-1β,IL-6 and TNF-α in group L compared with those at 1 day before surgery(P＞0.05),there were no significant differences in the levels of serum IL-1β,IL-6 and TNF-α among the three groups at 3 days after surgery(P＞0.05);Postoperative airway secretion was significantly reduced in group L and group M compared with group C(P＜0.05),and there was no significant difference between group L and group M(P＞0.05).The incidence of adverse reactions(dry mouth and dysuria)in group M was significantly higher than that in group L and group C(P＜0.05).Conclusion Low-dose(0.2 mg)glycopyrronium bromide is more appropriately recommended as a preoperative medication for elderly patients undergoing laparoscopic inguinal hernia repair surgery,and increased dosage is associated with an increase in adverse reactions and POCD.

Objective:The purpose of this study is to use the next-generation sequencing (NGS) technology platform to detect the methylation rate of phosphatase and tensin homolog deleted on chromosome ten ( PTEN) promoter region in hepatocellular carcinoma (HCC) tissue samples, and to analyze the clinical significance of its correlation with the prognosis of patients receiving sorafenib treatment. Methods:The 52 pairs of tumor tissue and para-cancerous tissue samples from HCC patients treated with sorafenib alone, which were collected and preserved in the Liver Tumor Diagnosis and Research Center of the former 302 Hospital of the People′s Liberation Army by the National Natural Science Foundation of China Youth Project with the project batch number 81702986 in 2018, were extracted total DNA from the samples. Then the DNA samples were treated with bisulfite and specific primers were designed to amplify the PTEN promoter region. Finally, the amplified products were analyzed by second-generation sequencing. In the analysis of clinical significance of PTEN methylation, log-rank statistical analysis was used to calculate whether there was a statistical difference in survival between the patient groups. Results:The methylation rate of PTEN promoter region in tumor tissues (29.17%±9.58%) was significantly higher than that in paracancer tissues (4.17%±2.86%)( t=19.970, P<0.05). At the same time, in HCC tissues, the methylation rate of the PTEN promoter region is negatively correlated with its expression ( F=47.270, P<0.000 1; Y=-1 800× X+38.03), and the PTEN methylation rate is negatively correlated with the prognosis of patients receiving the molecularly targeted drug Sorafenib (χ2=4.313, P<0.05). Conclusion:This study successfully established a new method for detecting methylation in the promoter region of PTEN, and the methylation rate of PTEN can be used as one of the targets of HCC diagnosis and targeted therapy.

Objective:The purpose of this study is to use the next-generation sequencing (NGS) technology platform to detect the methylation rate of phosphatase and tensin homolog deleted on chromosome ten ( PTEN) promoter region in hepatocellular carcinoma (HCC) tissue samples, and to analyze the clinical significance of its correlation with the prognosis of patients receiving sorafenib treatment. Methods:The 52 pairs of tumor tissue and para-cancerous tissue samples from HCC patients treated with sorafenib alone, which were collected and preserved in the Liver Tumor Diagnosis and Research Center of the former 302 Hospital of the People′s Liberation Army by the National Natural Science Foundation of China Youth Project with the project batch number 81702986 in 2018, were extracted total DNA from the samples. Then the DNA samples were treated with bisulfite and specific primers were designed to amplify the PTEN promoter region. Finally, the amplified products were analyzed by second-generation sequencing. In the analysis of clinical significance of PTEN methylation, log-rank statistical analysis was used to calculate whether there was a statistical difference in survival between the patient groups. Results:The methylation rate of PTEN promoter region in tumor tissues (29.17%±9.58%) was significantly higher than that in paracancer tissues (4.17%±2.86%)( t=19.970, P<0.05). At the same time, in HCC tissues, the methylation rate of the PTEN promoter region is negatively correlated with its expression ( F=47.270, P<0.000 1; Y=-1 800× X+38.03), and the PTEN methylation rate is negatively correlated with the prognosis of patients receiving the molecularly targeted drug Sorafenib (χ2=4.313, P<0.05). Conclusion:This study successfully established a new method for detecting methylation in the promoter region of PTEN, and the methylation rate of PTEN can be used as one of the targets of HCC diagnosis and targeted therapy.