Background and purpose:Previous studies have shown that Bubl was a critical component of the spindle checkpoint.Paclitaxel sensitivity was considered to be dependent on the functionality of this spindle checkpoint.This study investigated the effects of pEGFP-Bubl-shRNA plasmid stably transfected into the cell cycle and its sensitivity in human ovarian cancer cell line A2780.Methods:After the pEGFP-Bubl-shRNA plasmid and empty plasmids were constructed.they were transfected into A2780 cells by the Lipofectamine 2000~(TM).The nontransfected cells were the control.RT-PCR and Western blotting were used to determine the target gene and protein expression.The rate of proliferation inhibition was tested by an MTT assay,apoptosis and cell cycles were determined by flow cytometry,and the mitotic index was determined bv Hoechst33342 dye.Results:RT-PCR and Western blotting showed that pEGFP-Bubl-shRNA/A2780 group displayed a low expression of Bubl compared to the A2780 and pEGFP-Cl/A2780 group(P＜0.05).The sensitivity of the pEGFP-Bubl-shRNA/A2780 group was significantly lower than the non-transfccted and pEGFP-Cl/A2780 cells(P＜0.05).Flow cytometry revealed that the rates of G2/M phase and apoptosis were significantly lower in the pEGFP-Bubl-shRNA/A2780 group than those in the control group (P＜0.05).Conclusion:Bubl plays an important role in the paclitaxel treatment.A down-regulation of Bubl could reduce the drug sensitivity and rate of G_2/M cells in human ovarian cancer cell line A2780.

Dust ; Humans ; Silicon Dioxide ; adverse effects ; Silicosis ; etiology

Objective:To assess the prevalence of depressive symptoms in type 2 diabetic patients with hypertension.Methods:A systematic electronic search was conducted in CNKI,Wanfang database,SinoMed,Pubmed and Embase.Literatures that reported the depressive symptoms status of type 2 diabetic patients with hypertension were included.All patients were divided into case group and control group,according to with or without hypertension.Meta-analysis was performed with Statal2.0 software after data extraction and the quality of studies assessment.Results:A total of 11 articles involving 8031 type 2 diabetic patients were included in the meta-analysis,including 4711 patients in case group and 3320 patients in control group.The pooled prevalence of depressive symptoms in case group was 43.5％ (95％ CI:26.1％-60.8％).The pooled prevalence of depressive symptoms in control group was 34.0％ (95％ CI:19.6％-48.5％).The pooled odds ratio was 1.32 (95％ CI:1.05-1.67,P ＜0.05).Conclusion:Hypertension increases the risk of developing depressive symptoms in type 2 diabetic patients.Therefore,medical staff should pay more attention to the mental health of this population.

Objective To investigate changes in plasma levels of D-dimer in patients with prostate hyperplasia(BPH)complicat-ed with acute urinary retention(AUR),and to explore its clinical significance.Methods 102 cases of patients with prostate hyper-plasia complicated with acute urinary retention,treated in this hospital from July 2013 to December 2014,were selected and divided into the simple BPH group(56 cases)and BPH complicated with AUR(BPH+AUR)group.The plasma levels of D-dimer of these patients were detected and comparatively analysed.Results Compared with the simple BPH group,the plasma levels of D-dimer of patients in the BPH+AUR group was increased,had statistically significant difference.(P <0.01).Conclusion The plasma levels of D-dimer in patients with BPH complicated with AUR increase significantly,which indicates that patients are with secondary in-creased fibrinolytic activity.Measures should be taken to relieve urinary retention as soon as possible,so as to protect the vascular endothelial function of patients and reduce the incidence of complications.

Objective:To explore the validity of the effective arterial elastance (Ea) before and after fluid challenge in evaluating the fluid challenge in septic shock patients.Methods:A retrospective study was conducted in the medical intensive care unit (MICU) of Peking Union Medical College Hospital from October 2016 to October 2020. 116 septic shock patients were enrolled. All patients received fluid challenge by 500 mL Gelatin or normal saline under invasive hemodynamic monitoring. Heart rate (HR), mean arterial pressure (MAP), cardiac output (CO) and other hemodynamic variables were collected at 10 minutes before and immediately after fluid challenge. An increase in CO greater than 10% after fluid challenge was defined as the positive preload responsiveness, as well as the definition of positive pressure responsiveness was an increase in MAP greater than 10%. Receiver operating characteristic curves (ROC curves) were established to evaluate the predictive abilities of baseline Ea and other arterial load indices in detecting the preload responders and pressure responders. The correlation of the baseline Ea with CO changes after fluid challenge as well as MAP changes were tested by Pearson correlation analysis. Patients with positive preload responsiveness were divided into two groups according to the pressure responsiveness. The changes in Ea and other arterial load indices were analyzed.Results:A total of 116 patients were finally analyzed. Sixty-three patients were preload responders and 53 patients were preload non-responders. There was no significant difference in demographics and baseline physical variables between the two groups. Ea in preload responders was higher than that in preload non-responders (mmHg/mL: 2.51±1.08 vs. 1.87±0.68, P < 0.01). ROC curve analysis showed that the baseline Ea could predict the preload responsiveness at an area under ROC curve (AUC) = 0.71 [95% confidence interval (95% CI) was 0.62-0.81, P < 0.001]. The cut-off value was 1.97 mmHg/mL with a sensitivity of 71.4% and a specificity of 60.4%. The baseline Ea did not present the predictive ability to detect the pressure responders and pressure non-responders (AUC = 0.52, 95% CI was 0.41-0.63, P = 0.73). Pearson correlation analysis showed that the changes in CO after fluid challenge was moderately correlated to the baseline Ea ( r = 0.47, P < 0.001), meanwhile a weak positive correlation between the changes in MAP and baseline Ea was found ( r = 0.20, P = 0.03). In preload responders, 27 (42.9%) of 63 patients were pressure responders and 36 (57.1%) patients were pressure non-responders. No statistical difference was found in the baseline Ea or other arterial load indices between the two groups. Fluid challenge decreased Ea both in pressure non-responders and pressure responders (mmHg/mL: 2.13±0.94 vs. 2.51±1.08, P < 0.01; 2.47±1.18 vs. 2.69±1.30, P < 0.05). Moreover, the changes in CO and changes in MAP were strongly correlated with the changes in Ea ( r values were -0.50 and 0.58, respectively, both P < 0.001). Conclusions:The Ea > 1.97 mmHg/mL before fluid challenge could predict fluid responsiveness in septic shock patients. The baseline Ea was not able to predict the subsequent changes in arterial pressure through fluid challenge. A significant decrease in Ea inducing by fluid administration explained why patients increased their CO without improving blood pressure.

OBJECTIVETo determine whether a novel compound, 3-(4-bromophenyl)-2-(ethyl sulfonyl)-6-methylquinoxaline 1, 4-dioxide (Q39), induces apoptosis in human leukemia cell line k562 in hypoxic environment.

METHODSMTT assay was used to determine the 50% inhibitory concentrations (IC50). Flow cytometry and DAPI staining were employed to determine the apoptosis; JC-1 staining was used to determine mitochondria membrane potential (DeltaPsim); Western-blotting was used to determine protein expression of procaspase-3, cleaved caspase-3, PARP, Bax, Bcl-2 and HIF-1alpha.

RESULTSIn hypoxic environment, Q39 exerted higher antiproliferative activity in K562 cells, and the IC50 value was (0.21+/- 0.05) micromol/L. The apoptotic phenomenon was observed at 6 h after cells exposed to Q39, and apoptotic body emerged as exposure time increased. After K562 cells were incubated with Q39 for 0, 6, 12 and 24 h, the ratio of apoptotic cells was 2.8%, 3.2%, 5.9% and 19.2%, respectively. By fluorescence stain assay, an significant Delta Psim loss in K562 cells induced by Q39 was shown in a time-dependent manner. Western blot assay demonstrated that Q39 decreased the protein expression of Bcl-2, procaspase-3, and HIF-1alpha, meanwhile increased protein expression of Bax and cleaved caspase-3, and induced the cleavage of PARP.

CONCLUSIONSThe novel compound Q39 exhibits great anticancer activity against K562 cells in hypoxic environment. Q39 can downregulate the protein expression of HIF-1alpha, and regulate the apoptosis-related protein expression to cause a drop of DeltaPsim, suggesting that mitochondria and HIF-1alpha pathway might be involved in the antiproliferative effect of Q39.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Caspase 3 ; metabolism ; Cell Hypoxia ; Flow Cytometry ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; K562 Cells ; Membrane Potential, Mitochondrial ; drug effects ; Poly(ADP-ribose) Polymerases ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Quinoxalines ; pharmacology ; bcl-2-Associated X Protein ; metabolism

Objective To investigate the neurotoxic effects of different concentrations of tetracaine and ropivacaine on the brachial plexus nerve in rats.Methods Forty-eight male Sprague-Dawley rats,weighing 410-430 g,were randomly divided into 8 groups (n =6 each):normal saline group (group NS),0.25％,0.50％ and 1.00％ tetracaine groups (groups T1-3 ),and 0.25％,0.50％,1.00％ and 2.00％ ropivacaine groups (groups R1-4 ).The rats received injection of normal saline 1.0 ml,0.25％,0.50％ and 1.00％ tetracaine 0.5 ml,0.25％,0.50％,and 1.00％ ropivacaine 1.0 ml and 2.00％ ropivacaine 0.5 ml in groups NS,T1-3 and R1-4 respectively through one side of the axillary sheath.The other side of the axillary sheath served as control side.Five days later,compound action potential and nerve conduction velocity (NCV) of the brachial plexus nerve were measured.Tne brachial plexus nerve was obtained as the specimen for microscopic examination with light and transmission electron microscope.Results Compared with the control side and group NS,the compound action potential and NCV of the brachial plexus nerve were significantly decreased in groups T2,3 and R3,4 ( P ＜ 0.05 ).The compound action potential and NCV of the brachial plexus nerve were gradually decreased with the increasing concentrations of tetracaine in groups T1 3 ( P ＜ 0.05 ).The compound action potential and NCV of the brachial plexus nerve were significantly decreased in group R4 as compared with groups R1-3 (P ＜ 0.05).The microscopic examination showed that the pathologic changes were more severe in groups T2,3 and R3,4 than those on the control side and than in group NS.Conclusion 0.50％ and 1.00％ tetracaine,and 1.00％ and 2.00％ ropivacaine can result in pathologic damage to the brachial plexus nerve in rats and the degree of damage is related to the concentration.

ObjectiveTo investigate the incidence of bacterial meningitis in patients who underwent intracranial neurosurgery and to explore the pathogen distribution and related risk factors.MethodsThe patients were selected by cluster systematic sampling method from the patient population who underwent intracranial neurosurgery for at least one time in Huashan Hospital affiliated to Fudan University in 2008.Chi-square test and Logistic regression model were used to analyze the data.ResultsA total of 1165 patients were included in this study.Seventy-nine of them were diagnosed with bacterial meningitis,while eight of them bad positive results for bacterial culture,including 3 cases of Acinetobacter baumannii,and 1 case each of Acinetobacter lwoffii,Klebsiella pneumoniae,Staphylococcus epidermidis,Enterococcus and Streptococcus intermedius. The common neurosurgical diseases with high incidence of postoperative bacterial meningitis were neurilemmoma 15.85％(13/82),glioma 12.21％ (21/172) and hydrocephalus 10.34％ (3/29).Logistic regression analysis identified male,implant,enteral nutrition,external ventricular drainage and wound negative pressure drainage as independent risk factors for postoperative bacterial meningitis.ConclusionsPostoperative central nervous system infection is the most common and severe complication after the intracranial neurosurgery.The results highlight the importance of improving pathogen detection rate and enhancing the prevention in high risk patient populations.

Recent evidence has suggested that Akt2 plays an important role in the protection of cells from paclitaxel (PTX)-induced apoptosis and control of the cell cycle. In addition, some scholars suggested that the PTX sensitivity depends on a functional spindle assembly checkpoint. In the present study, we investigated the role of the Akt2/Bub1 cross-talking in apoptosis and cell cycle after exposure of the A2780 ovarian cancer cells to paclitaxel (PTX). Recombinant expression plasmid WT-Akt2 was transfected into A2780 cells by lipofectamine2000, and then the expression level of Akt2 gene was detected by using RT-PCR and Western blotting. Cell apoptosis and cell cycle were detected by flow cytometry and Hoechst 33342 staining after treatment with PTX. Moreover, we compared the expression level of Bub1 in different groups by Western blotting. Our study showed that up-regulation of Akt2 contributed to A2780 ovarian cancer cells overriding PTX-induced G(2)/M arrest, and inhibited Bub1 expression. Our findings might shed light on the molecular mechanism of PTX-induced resistance in ovarian cancer and help develop novel anti-neoplastic strategies.

Objective To study the early protective effect of NADPH on human umbilical vein endothelial cells (HUVECs) and the expression of occludin and MMP9 induced by oxygen glucose deprivation and reoxygenation (OGD/R). Methods HUVECs were cultured and divided into blank control group, OGD/R group and OGD/R+NADPH 20 μmol/L group. The proliferation of HUVECs after treatment was detected by CCK-8 assay. The cytotoxicity was detected by LDH release assay. The morphological changes of HUVECs were observed by inverted microscope. Superoxide dismutase (SOD MDA) activity and malondialdehyde (MDA) were detected by commercially available kit. The expressions of occludin and MMP9 were detected by Western blot assay. Results Compared with the OGD/R, NADPH enhanced the cell viability significantly (P<0.05), reduced the release of LDH (P<0.05), promoted the maintance of HUVECs morphology, reduced MDA generation (P<0.05) and increased SOD activity (P<0.05). Following OGD/R,the treatment of NADPH can inhibit MMP9 level (P<0.05) and promote the recovery of occludin level (P<0.05). Conclusion NADPH can protect HUVECs from the damage induced by OGD/R by reducing oxidative stress and regulating the expressions of MMP9 and occludin.