Objective To compare the difference in quercetin against oxidative stress response in mouse and in NIH-3T3 cells before and after H2O2 treatment,to explore the underlying mechanism for the quercetin antioxidant.Methods The cultured NIH-3T3 cells were randomly divided into 4 groups: quercetin(Q) pre-protective group(Qb) firstly treated with quercetin for 24 h followed by incubation with H2O2 for 30 min;post-protective group(Qa) treated with H2O2 for 30 min followed by incubation with quercetin for 24 h;H2O2 group(H2O2) after exposure to H2O2 for 30 min,incubated with DMEM medium and the control group(C) only cultured with DMEM medium.The survival rate and apoptotic rate were detected respectively with MTT and TUNEL in NIH-3T3 cell sus-pension samples.The expression of cyclin D1,PTEN,NF-?B,HSP-70,BCl-2,BAX and caspase-3 were examined with immunocytochemistry and immunoblotting.Besides,20 Wistar rats were divided into control group and experimental group,the latter was given with quercetin in the doze of 0.13 mmol/kg.The levels of T-AOC,SOD,GSH-Px,GSH,MDA,NOS and NO2-/NO3-were detected both in the cleaved NIH-3T3 cells and in the plasma from both experimental and control animals prior to and post-1 h,2 h and after 24 h.Results When the Qb group was compared with H2O2 or Qa group,the survival rate was higher and the apoptotic rate was lower.When the H2O2 group was compared with C group,the expression of cyclin D1、PTEN or BCl-2 was down-regulated;while that of BAX、HSP-70、NF-?B or caspase-3 was up-regulated;the level of T-AOC,SOD,GSH-Px or GSH was decreased;that of NOS、NO2-/NO3-or MDA enhanced in the cleft NIH-3T3 cells.When the plasma level of the anti-oxidative enzyme system prior to-compared with post-1h and 2h-treatment with Q,the level of T-AOC,SOD,GSH-Px and GSH,especially the former two,were higher;MDA,lower;NOS or NO2-/NO3-promoted.However,the above parameters basically became normal 24 h after treatment with Q.Conclusion Quercetin down-regulates the promoted expression of HSP70,NOS,NO2-/NO3-and NF-?B etc.in H2O2-treatment NIH-3T3 cells.Qb could reverse the H2O2 damage effects more markedly.Moreover,the quercetin exerts anti-oxidant protective effect through modulating the anti-oxidative enzyme system both in vivo and in vitro.However,based on the cell heterogeneity in none-or pre/post-H2O2-treatment state,a difference in quercetin antioxidant response is noted.

Objective To study the expression of SGK1 in T lymphocytes from pediatric asthma,and the effect of SGK1 on the differentiation of T cells,also to explore the function of SGK1 regulating the differen-tiation of T subset in pediatric asthma. Methods Twenty-eight children with asthma were recruited in Xi′an children′s hospital and divided into moderate group and severe group according to diagnostic guideline of asth-ma. The serum levels of IL-4,IL-13 and IL-17A were analyzed by ELISA. The CD4 +T cells from PBMC and na?ve T cells were selected using magnetic beads. Na?ve T cells were differentiated in vitro under cytokines. SGK1 expression were analyzed with Real-time PCR. The ability of Th2 and Th17 on secreting IL-4 and IL-17A were detected after SGK1 was inhibited by siRNA. In vivo,shRNA-SGK1 Na?ve T cells were transferred into the mice asthma models by intravenous injection. The airway inflammation were observed in shRNA-SGK1 Na?ve T models. Results Compared with healthy children,the serum levels of IL-4、IL-13 and IL-17A increased signifi-cantly in the children with asthma. Importantly,the levels of these three cytokines were much higher with the de-velopment of asthma. SGK1 were up-regulated remarkably in CD4 +T cells from the children with asthma and were positively correlated with IL-13 and IL-17A. Besides,SGK1 expression increased in the differentiated Th2 and Th17 in vitro,but had no change in the differentiated Th1. The levels of IL-4 and IL-17A associated with Th2 and Th17 decreased after SGK1 was inhibited by siRNA. Similarly,In vivo,the serum levels of IL-13 and IL-17A and airway inflammation were reduced in shRNA-SGK1 Na?ve T models. Conclusion The over-expres-sion of SGK1 in pediatric asthma enhances the asthma progress by promoting the differentiation of T subsets.

Tumor associate macrophages ( TAMs) play a significant role in the interaction of tumor inflam-mative microenvironment and tumor cells .TAMs originate from monocytic precursors ,recruiting into tumor tissue by colony stimulating factor ( CSF) .This review summarized that TAMs promote tumor progression and metastasis though angiogenesis ,lymphogenesis , immunosuppression , matrix remodeling and affecting cancer stem cells .The article pointed that targeting TAMs is a new strategy for future tumor therapy .

Objective To explore the effects of natural avocado oil and olive oil on the proliferation and differentiation of HaCaT cells. Methods MTT assay was performed to determine the optimal work concentration of avocado oil and olive oil. Cultured HaCaT cells were divided into 3 groups, i.e., avocado oil group treated with avocado oil of 3% (v/v), olive oil group treated with olive oil of 3% (v/v), and control group without any treatment. Immunocytochemistry and immuno-dot-blot method were used to detect the expressions of c-myc, mitogen-activated protein kinase ( MAPK ), nuclear factor ( NF)-κB, filaggrin, involucrin and keratin10 in HaCaT cells. Results As immunocytochemistry showed, the mean grey values (staining intensity) of c-myc,MAPK, and NF-κB in HaCaT cells were 131.4 ± 6.6,136.3 ± 4.5 and 134.3 ± 5.2 respectively in the avocado oil group, 121.1 ± 4.5, 107.9 ± 7.3 and 106.4 ± 5.4 respectively in the olive oil group, significantly higher than that in the control group (101.9 ± 8.9,91.4 ± 5.1 and 94.3 ± 7.0, respectively, all P＜ 0.05), and the avocado oil group was higher than the olive oil group in all the above parameters (all P ＜ 0.05). Increased expressions of filaggrin, involucrin and keratin 10 were observed in the avocado oil group and olive oil group compared with the control group (all P＜ 0.05), and in the olive oil group than in the avocado oil group (all P＜ 0.05).The mean grey values of these proteins obtained by immunocytochemisty were significantly correlated with those obtained by immuno-dot-blot method in avocado oil group (r = 0.94, P ＜ 0.01 ) and olive oil group (r=0.97, P ＜ 0.01 ). Conclusions Certain concentrations of avocado oil and olive oil can promote the proliferation and differentiation of HaCaT cells; avocado oil is more capable to accelerate their growth and proliferation, and olive oil to enhance their differentiation.

BACKGROUND: Exercise has been proved to accelerate the proliferation of intervertebral disc cells and extracellular matrix production in healthy rats. For the degenerative intervertebral disc, whether exercise also has positive effects on its cell proliferation, extracellular matrix production or pain relief remains unclear. OBJECTIVE: To investigate the effect of exercise on the extracellular matrix production in a rat model of intervertebral disc degeneration.METHODS: A rat model of intervertebral disc degeneration was prepared by Freund's complete adjuvant injection into the intervertebral disc at L5-6 levels. Then, the model rats were allowed to have a rest for 2 weeks. All rats were then randomly divided into exercise and control groups. Rats in the exercise group were forced to run every day, while the controls allowed free activities in the cage. The behavioral tests were performed at 7, 14, 28, 42, 56 and 70 days after modeling; meanwhile, the intervertebral disc samples were collected used for alcian blue staining and immunohistochemical staining to detect the levels of proteoglycan, aggrecan and collagen type Ⅱ in the intervertebral disc cells, respectively.RESULTS AND CONCLUSION: Vocalization threshold on the rat back of punctured disc was significantly decreased, while grooming and wet-dog shaking were significantly increased at 7 days after modeling compared with the baseline (P < 0.05), suggesting that Freund's complete adjuvant injection successfully induces disc degeneration, hyperalgesia and abnormal behaviors. Further, the vocalization threshold and wet-dog shaking in the exercise group showed significant improvement compared with the control group after 14 days of exercise (P < 0.05), while the grooming was significantly reduced until the 28th day (P < 0.01), indicating that exercise can alleviate pain caused by disc degeneration in model rats. At 21 days after modeling, the levels of proteoglycan, aggrecan and collagen type Ⅱ in the nucleus pulposus and annulus fibrosus were significantly decreased compared with the baseline (P < 0.01), indicating the occurrence of disc degeneration. After 14 days of training, the levels of proteoglycan, aggrecan, and collagen type Ⅱ in the nucleus pulposus and annulus fibrosus in the exercise group were significantly increased compared with the control group (P < 0.01). Moreover, after 8-week exercise, the level of proteoglycan in the nucleus pulposus and annulus fibrosus in the exercise group was increased by 4-5 times compared with the control group, and levels of aggrecan and collagen type Ⅱ in the nucleus pulposus in the exercise group also was increased by 3-4 times compared with the control group. To conclude, exercise can promote extracellular matrix increased by production by increasing the levels of proteoglycan, aggrecan, and collagen type II in the degenerative intervertebral disc.

Objective:To investigate the effects of bezafibrate (BF) on the activation,proliferation and differentiation of CD4+T cells from primary biliary cirrhosis ( PBC) patients and to elucidate the mechanisms for the immunosuppressive effects of BF and to further provide experience basis for BF target therapy PBC.Methods:PBMCs were isolated from PBC patients then CD 4+T cells were selected by MACS, and stimulated with anti-CD3, anti-CD28, in the presence of different concentration of BF.The cytokines were measured by ELISA,and the activation,proliferation and differentiation of CD4+T cells were analyzed by flow cytometry.Results:(1) BF could inhibit the activation of CD 4+T cells in PBC patients.(2) BF could inhibit the proliferation of CD 4+T cells in PBC patients in a dose-dependent manner (P<0.05).(3)BF could down-regulation IFN-γand IL-17 production of CD4+T cells in a dose-dependent manner ( P<0.05 ).Conclusion: BF could inhibit immune responses of PBC patients by suppressing CD 4+T cells activation;proliferation and cytokine production.

Objective To study the profile of peripheral blood cortisol concentration in patients with hepatitis B virus (HBV)-related acute on chronic liver failure (ACLF) and its association with disease severity and predictive value on prognosis.Methods Forty-five patients with HBV-related ACLF,including 15 patients in early-stage,15 in medium stage and 15 in end-stage; and 15 patients with severe chronic hepatitis B (CHB) were also enrolled.Peripheral blood cortisol concentration was tested by chemiluminesence immunoassay.SPSS version 18.0 was used to compare peripheral blood cortisol concentration among different groups and analyze its correlation with prothrombin activity (PTA),total bilirubin (TBil),albumin (Alb),alanine aminotransferase (ALT),aspartate aminotransferase (AST),HBV DNA,international normalized ratio (INR) and model for end-stage liver disease (MELD) scores.The independent risk factors of prognosis in patients with HBV-related ACLF were determined by performing Logistic regression analysis.Results The concentrations of peripheral blood cortisol in severe CHB group,early stage ACLF group,medium stage ACLF group and end-stage ACLF group were (595.6±114.0),(496.0±87.2),(303.9±81.1),and (183.8±71.8) nmol/L,respectively.A decreasing trend was observed and the differences among groups were statistically significant (F =63.93,P ＜ 0.01).Peripheral blood cortisol concentration were significantly different among subgroups of patients with MELD＜30,followed by 30-40 and ＞40(F 9.01,P＜0.01).Peripheral blood cortisol concentration in patients with ACLF was positively correlated with PTA level (r=0.83,P＜0.01),and inversely correlated with TBil (r =-0.34,P＜0.05),MELDscore (r =-0.60,P＜0.01),AST/ALT (r =-0.35,P＜0.05),and INR (r=-0.59,P＜0.01).Association with Alb,ALT,AST and HBV DNA was not observed.According to multivariate Logistic regression analysis,MELD score,cortisol level,Alb and total cholesterol were independent risk factors of prognosis for patients with ACLF.Serum cortisol concentration in survival group of HBV-related ACLF was higher than that in death group,while the death group exhibited a gradually decreasing trend.Conclusions The peripheral blood cortisol concentration decreases in patients with HBV-related ACLF,which is related to the degree of liver dysfunction,disease severity and prognosis of patients.Moreover,the lower level of cortisol concentration indicates poor short-term prognosis in patients with HBV related ACLF.

Objective:To explore the impact of completion rates of 3-hour and 6-hour sepsis bundle therapy on prognosis of patients with septic shock in Prefecture-level grade A hospitals, and analyze the risk factors for prognosis.Methods:A retrospective analysis was conducted to patients with septic shock in the intensive care unit (ICU) of Liaocheng People's Hospital, Shandong Province from January 1, 2020 to December 31, 2021. The data of gender, age, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ), sequential organ failure assessment (SOFA), sites of infection, pathogenic microorganisms, completion rates of 3-hour and 6-hour sepsis bundle therapy, 28-day prognosis were collected. Logistic regression analysis was used to identify risk factors for patients' mortality at 28-day.Results:① Among 159 patients with septic shock, 93 survived and 66 died with 28-day. There were no significant differences in gender and age between the survival group and death group. Compared with the survival group, APACHE Ⅱ score and SOFA score were significantly higher in the death group [APACHE Ⅱ score: 26.85±5.04 vs. 20.67±4.29, SOFA score: 12.86±3.02 vs. 9.37±2.51, both P < 0.05]. ② Sites of infection in the 159 patients: 47 cases were abdominal infection (29.6%), 36 case were bloodstream infection (22.6%), 31 cases were pulmonary infection (19.5%), 16 cases were soft tissue infection (10.1%), 13 cases were urinary tract infection (8.2%), 12 cases were biliary tract infection (7.5%), and 4 cases were other sites infection (2.5%). Pathogens were found in 128 cases and the positive rate was 80.5%, including 90 cases of Gram-negative (G -) bacilli (56.6%), 27 cases of Gram-positive (G +) cocci (17.0%) and 11 cases of fungi (6.9%). The top three pathogenic bacteria were Escherichia coli (49 cases, 30.8%), Klebsiella pneumoniae (21 cases, 13.2%) and Staphylococcus aureus (15 cases, 9.4%). The differences were not statistically significant. ③ Among the 159 patients, 101 cases completed 3-hour sepsis bundle therapy (63.5%), including 67 cases (72.0%) in survival group and 34 cases (51.5%) in death group; 106 cases completed 6-hour sepsis bundle therapy (66.7%), including 70 cases (75.3%) in survival group and 36 cases (54.5%) in death group. The differences between the two groups were statistically significant (all P < 0.05). ④ The factors (APACHE Ⅱ score, SOFA score and completion rate of 3-hour and 6-hour sepsis bundle therapy) affecting the prognosis in the univariate analysis were included in the binary Logistic regression analysis, and the results showed that the APACHE Ⅱ score, SOFA score, completion rate of 3-hour sepsis bundle therapy were independent risk factors affecting mortality within 28-day [odds ratio ( OR) was 1.216, 1.303, 0.402, all P < 0.05]. Conclusions:The higher APACHE Ⅱ score and SOFA score in septic shock, the worse the prognosis. Improving the completion rates of 3-hour and 6-hour bundle therapy especially the completion rate of 3-hour bundle therapy can reduce the mortality of patients and improve the prognosis.

During the prevention and control of coronavirus disease 2019 (COVID-19) epidemic, our hospital was a designated hospital for the treatment of COVID-19, and established the infection control system in the department of anesthesiology according to the clinical practice. The relevant staff of our hospital strictly followed the rules and procedures for operation, and no staff members were found to be infected by emergency surgical anesthesia performed in COVID-19 patients. The experience is summarized as follows: timely establishment and unified command of the emergency infection control management team in the department of anesthesiology; scientific formulation and dynamic improvement in comprehensive, systematic and feasible infection control strategies, norms and procedures; clear division of responsibilities and adequate management and supervision of infection control management team members; carrying out strict training of infection control, establishing a good awareness of infection control in the operating room, and implementing standard infection control procedures from the medical staff of the department to the cleaning staff.

OBJECTIVETo study the molecular mechanism of tetramethylpyrazine to induce human promyelocytic HL-60 leukemia cells differentiation.

METHODThe cell proliferation was determined by MTT. The differentiation of the cells was detected by NBT reduction test. Cellular morphology was observed by Wright's staining. Cell cycle distribution and the distribution of CD11b, CD14 were detected by flow cytometry. Then RT-PCR and Western blot assay were employed to detect the expressions of c-myc, p27, CDK2 and cyclinE1 in HL-60 cells after exposure to TMP.

RESULTTMP inhibited the proliferation in a dose and time dependent manner. TMP at the concentration of 200 mg x L(-1) to 300 mg x L(-1) induced unterminal differentiation of HL-60 cell and synergistically blocked the cell cycle progression of HL-60 cells in G0/G1 phase. The expression of c-myc was down-regulated as well as the protein expression of cyclin E and CDK2, while the mRNA and protein expression of P27 were remarkably up-regulated.

CONCLUSIONSmall doses of TMP induces differentiation of HL-60 cells throughout the cell cyde, as detected by a slower rate of accumulation in G0/G1, possibly by regulating the expression and activity of G1/S phase-related molecules.

Cell Cycle Proteins ; genetics ; metabolism ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Gene Expression Regulation, Leukemic ; drug effects ; HL-60 Cells ; Humans ; Leukemia, Promyelocytic, Acute ; drug therapy ; genetics ; metabolism ; physiopathology ; Pyrazines ; pharmacology