Objective To investigate whether homocysteine(Hcy) induces apoptosis of endothelial cells via a pathway involving caspases3 and whether simvastatin antagonizes the proapoptotic effects of Hcy by regulating c-IAP. Methods Human umbilical vein endothelial cells(HUVEC) were treated with Hcy,with or without simvastatin,for 24 h.Cell apoptosis was evaluated by Annexin V staining and flow cytometery,as well as TUNEL.The mRNA and protein levels of caspase3,c-IAP-1 and c-IAP-2 were analyzed by RT-PCR and Western blot,respectively. Results Treatment with both low(0.5 mmol/L) and high(3.0 mmol/L) concentrations of Hcy-induced HUVEC apoptosis was accompanied by an increased level of caspase3 expression and activation,together with decreased c-IAP-1 and c-IAP-2 level.Simvastatin upregulated c-IAP-1 and c-IAP-2 expression while attenuated Hcy-induced apoptosis and caspase3 activation. Conclusion Hcy may induce HUVEC apoptosis via a pathway involving caspase3,which can be partially antagonized by simvastatin,possibly through upregulated c-IAP-1 and c-IAP-2 expression.

Objective - （ ） To investigate the effects of nuclear factor erythroid 2 related factor 2 NRF2 on the oxidative stress （ ） Methods ） ，， induced by trichloromethane TCM in human normal hepatocyte L02 cells. i L02 cells were stimulated with 1 2 ， ， ， （ ）， 4 8 12 16 and 20 mmol/L TCM solution dissolved in dimethyl sulfoxide and the control group and blank group were set ， - ， up. After culturing for 24 hours the cell viability was detected by CCK 8 colorimetric method and the concentration of TCM ） -， - stimulation was screened. ii L02 cells in logarithmic growth phase were randomly divided into control group and low medium - ， ， ， and high dose groups. After 24 hours of exposure to 0 4 8 and 12 mmol/L TCM the cells were collected. The activity of （ ）， （ ）， （ - ） （ ） superoxide dismutase SOD catalase CAT glutathione peroxidase GSH Px and the level of malondialdehyde MDA NRF2， - （HO-1）， were detected by colorimetric analysis. The mRNA expression levels of heme oxygenase 1 glutamate cysteine （GCLC） （） （NQO1） - ligase catalytic subunit and NAD P H quinone dehydrogenase 1 were detected by real time fluorescence ， - ， polymerase chain reaction. The protein levels of NRF2 HO 1 GCLC and NQO1 were detected by Western blotting.Results ） ， ， ， ， i When the concentration of TCM was 4 8 12 16 and 20 mmol/L the survival rate of L02 cells decreased （ P ） ， ， significantly compared with the control group all <0.05 . The concentration of 0 4 8 and 12 mmol/L were selected as the ） ， - stimulation doses for subsequent experiments. ii Compared with the control group the activities of SOD and GSH Px in L02 （ P ） （ P ）， - cells in the three doses groups decreased all <0.05 and the levels of MAD increased all <0.05 with a dose effect - （P ）， relationship. The CAT activity of L02 cells in the medium dose group was lower than that in the control group <0.05 and the - （ P ） CAT activity of L02 cells in the high dose group was lower than that in the others three groups all <0.05 . Compared with the ， NRF2 - （P ），NRF2 control group the relative expression levels of mRNA in L02 cells in the low dose group decreased <0.05 - （P ）， NRF2 mRNA in L02 cells in the medium dose group increased <0.05 mRNA and NRF2 protein expression in L02 cells in （ P ） HO-1，GCLC， NQO1 ， the highdose group increased both <0.05 . The relative expression level of mRNA and GCLC NQO1 （ P ） protein expression in L02 cells in the three doses groups increased compared with the control group all <0.05 . The relative NRF2 - - - expression level of mRNA in L02 cells in the high dose group was higher than that in the low and medium dose groups （ P ）， - （P ）， both <0.05 and the relative expression of NRF2 protein was higher than that in the low dose group <0.05 but the HO-1 GCLC - - （ relative expression levels of and mRNA and HO 1 protein level were lower than those in the medium dose group all P ）Conclusion - <0.05 . TCM exposure can inhibit the proliferation of L02 cells by inducing oxidative stress with a dose effect ， relationship. In this process the antioxidant mechanism mediated by NRF2 was activated. The expression of antioxidant defense ， - ， and detoxification related target genes downstream of NRF2 signaling pathway was activated and the expression of HO 1 - GCLC and NQO1 was up regulated to alleviate the oxidative damage caused by TCM.

To explore the antibacterial activity and mechanism of total alkaloids and berberine from Coptidis Rhizoma on Aeromonas hydrophila, and determine the effect of total alkaloids and berberine from Coptidis Rhizoma on minimum inhibitory concentrations, permeability and fluidity of cell membrane, conformation of membrane proteins and virulence factors of A. hydrophila. The results showed that both total alkaloids and berberine from Coptidis Rhizoma had antibacterial activities on A. hydrophila, with minimum inhibitory concentrations of 62.5 and 125 mg · L(-1), respectively. Total alkaloids and berberine from Coptidis Rhizoma could increase the fluidity of membrane, change the conformation of membrane porteins and increase the permeability of bacteria membrane by 24.52% and 19.66%, respectively. Besides, total alkaloids and berberine from Coptidis Rhizoma significantly decreased the hemolysis of exotoxin and the mRNA expressions of aerA and hlyA (P < 0.05, P < 0.01), the secretion of endotoxin and the mRNA expression of LpxC (P < 0.05, P < 0.01). The results suggested that the antibacterial activity of total alkaloids and berberine from Coptidis Rhizoma on A. hydrophila may be related to the bacteria membrane injury. They inhibited the bacterial growth by increasing membrane lipid fluidity and changing conformation of membrane proteins, and reduced the secretion of virulence factors of A. hydrophila to weaken the pathogenicity.
Aeromonas hydrophila ; drug effects ; genetics ; metabolism ; Alkaloids ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Bacterial Proteins ; genetics ; metabolism ; Bacterial Toxins ; biosynthesis ; Berberine ; pharmacology ; Cell Membrane ; drug effects ; genetics ; metabolism ; Coptis ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Membrane Fluidity ; drug effects ; Rhizome ; chemistry

OBJECTIVETo investigate the effects of PLK1 gene silence by short hairpin RNA (shRNA) on PLK1 expression and apoptosis in K562 cells, and explore the role of PLK1 in the pathogenesis of leukemia.

METHODSThe shRNA fragment targeting at 1416-1436 bp of PLK1 mRNA was synthesized and cloned into pEGFP-H1 vector, named as pEGFP-H1/PLK1. The empty control, pEGFP-H1 and pEGFP-H1/PLK1 were transfected into K562 cells respectively via electroporation. 24 h or 48 h after transfection, gene and protein expression of PLK1 in the cells were assayed by RT-PCR and Western blot analysis respectively, cells viability by MTT assay, caspase-3 activity by colorimetry, cell cycle and apoptosis by FACS.

RESULTS24 and 48 h after transfection, PLK1 expression in K562 cells was 1.25 +/- 0.07 for control group, 0.52 +/- 0.04 and 0.25 +/- 0.02 for pEGFP-H1/PLK1 group, and 1.24 +/- 0.08 and 1.23 +/- 0.09 for pEGFP-H1 group respectively. The alteration status of PLK1 protein levels were similar to that of PLK mRNA levels. The apoptosis rate was (8.3 +/- 0.6)% in control group, (8.7 +/- 0.7)% in pEGFP-H1 group and (49.7 +/- 3.8)% and (82.3 +/- 6.9)% in pEGFP-H1/PKLK1 group at 24 and 48 h, respectively. In addition, cell fraction at G(2)/M phase was increased obviously compared with control and pEGFP-H1-transfected group.

CONCLUSIONThe constructed shRNA can remarkably inhibit PLK1 expression and transfected K562 cell proliferation, increase apoptosis and block cell-cycle, suggesting that PLK1 play important roles in apoptosis and cell-cycle control of leukemia cells.

Apoptosis ; genetics ; Cell Cycle ; Cell Cycle Proteins ; genetics ; metabolism ; Cell Proliferation ; Genetic Vectors ; Humans ; K562 Cells ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; genetics ; Transfection

OBJECTIVETo explore factors affecting occupational adaptability in nurses for offering basis to increase their occupational adaptability.

METHODSFive hundred and forty-five nurses were investigated with work ability index questionnaire and occupational stress instruments.

RESULTSThere were many risk factors affecting occupational adaptability in nurses. The main variables that influenced occupational adaptability included work-overtime, mental load, social support, physical environment, and job hazards. The social support was the factor increasing the occupational adaptability of the nurses (P < 0.01, OR = 0.912). Five factors including work overtime, mental load, social support, physical environment and job hazards were introduced in the Logistic equation. The established functions were: Logit (P) = -11.357 + 1.011x(1) + 0.335x(2) - 0.076x(3) + 0.260x(4) + 0.129x(5).

CONCLUSIONThere are many risk factors affecting occupational adaptability in nurses. Relevant measures should be taken to promote the occupational adaptability in nurses to reduce the risk factors.

Adaptation, Psychological ; Adolescent ; Adult ; Humans ; Logistic Models ; Middle Aged ; Nurses ; psychology ; Occupational Health ; Risk Factors ; Social Support ; Stress, Psychological ; epidemiology ; prevention & control ; Surveys and Questionnaires ; Workload ; psychology

OBJECTIVETo investigate the influence of enteral administration of carbachol on the intestinal dysfunction of both severely burn patients and rabbits with partial intestinal ischemia/reperfusion (I/R) injury.

METHODSSeventy-five white rabbits were inflicted with I/R injury and randomized into intestinal I/R (I, n=25), carbachol [C, n=25, with 3g/L carbachol (3 mg/kg) injection into duodenum 1 h after SMA occlusion] and sham operation (SO, n=25, with SMA isolation but no occlusion) groups, and 5 other as normal controls. The blood flow of intestinal mucosa was detected before and after SMA occlusion or admission of carbachol. Changes in diamine oxidase (DAO), D-lactate, xylopyranose absorption, blue dextran discharging time were measured at 2, 4, 6, 8, 24, 48, 72 h after SMA occlusion. In addition, eight severe burn patients with TBSA of 84 +/- 12% were enrolled in the study, and carbachol (15 microg/kg) was administered to patients when abdominal distension or bowel sound was lower than 2 times/min, then the number of abdominal distension and bowel sounds per minute were observed.

RESULTSThe blood flow in intestinal mucosa of rabbits without SMA occlusion was (102 +/- 5) PU, reduced to (48 +/- 6) PU after SMA occlusion, and increased to (77 +/- 3) PU after injection of carbachol. The plasma DAO activity and D-lactic acid content in I group began to increase 4 hours after SMA occlusion, and they reached the peak 24 hours after SMA occlusion (4.63 +/- 0.27 U/ml, 7.9 +/- 2.4 mg/L) , after that they decreased gradually, but still higher than the normal value (0.89 +/- 0.14 U/ml, 2.0 +/- 1.1 mg/L, P < 0.05). In carbachol group, data showed the same trends as that in intestine I/R group with lower values, while no obvious changes were in sham operation group (P > 0.05). The content of D-lactic decreased dramatically 2 hours after D-lactic administration in both I and C groups, increased 6 hours after SMA occlusion, then decreased gradually, but it in C group was always higher than normal values, and little fluctuation was in sham operation group. There was no blue dextran discharge 2 hours after SMA occlusion. The discharging distance increased 6 hours later, but it was obviously shorter than the normal value 24 hrs after operation (P < 0.05) , then it returned to normal 48 to 72 hrs after operation. In the C group, blue dextran discharge was found immediately after its injection, with obvious increase in the discharging distance to peak value (43 +/- 6 cm) 6 hours after injury, and returning to normal (28 +/- 3 cm) gradually. In severe burned patients, the bowel sounds was (1.6 +/- 1.1) per minutes before carbachol administration, then increased dramatically to (6.9 +/- 1.7) per minutes 10 mins after administration, reached to a higher level 30 minutes after administration (8.3 +/- 2.4 ) times/min, and it maintained to (6.1 +/- 1.3) times/min 1 hour after administration. Abdominal distension was ameliorated 2 hours after carbachol administration, six patients were able to defecate.

CONCLUSIONEnteral administration of Carbachol can increase the blood flow of intestine mucosa, help to improve the movement, absorption and barrier functions of intestine, and ameliorate intestinal dysfunction in patients with severe burns.

Adolescent ; Adult ; Animals ; Burns ; drug therapy ; physiopathology ; Carbachol ; therapeutic use ; Disease Models, Animal ; Female ; Humans ; Intestinal Mucosa ; blood supply ; metabolism ; Intestines ; physiopathology ; Male ; Rabbits ; Reperfusion Injury ; drug therapy ; physiopathology

BACKGROUNDPseudoaneurysms (PAs) are common vascular abnormalities predominantly arising from a disruption in the integrity of the arterial wall. The potential complications of PAs are usually unpredictable and carry high rates of morbidity and mortality. This paper presents our experience with various treatment strategies for PAs.

METHODSFifty-four patients with 55 PAs were diagnosed by non-invasive imaging examination. The etiology of PAs included trauma (33/55), infection (5/55), iatrogenic (6/55), and idiopathic (11/55). Different procedures including ultrasound (US)-guided compression, endovascular treatment, and surgery were performed depending on the location of PAs, size of the sac and neck, and characteristics of the donor artery. The methods of endovascular treatment included embolization of parent artery, the PA sac, or implantation of a stent-graft. Follow-up was performed using US or CT and ranged from 1 day to 24 months (average 16.7 months).

RESULTSIn all 54 patients, 3 patients with superficial PAs were treated by US-guided compression, while 44 patients with 45 PAs located in the head and neck (n = 20), viscera (n = 10) or extremities (n = 15) were treated by endovascular treatment. Nine patients with PAs located in the head and neck (n = 2) or extremities (n = 7) were treated by surgery. Among them, one patient underwent endovascular treatment combined with surgery and 1 was treated by surgery after unsuccessful US-guided compression. In the 3 patients treated with US-guided compression, 2 were successfully treated while the remaining patient required additional surgery. Primary technical success of endovascular management was 97.7% (43/44) and the cure rate was 95.5% (42/44). In the surgery group, 4 patients recovered well, 1 patient was cured by endovascular treatment combined with surgery, 2 cases underwent amputation, 1 patient died of multi-organ failure and 1 patient was paralysed.

CONCLUSIONSMinimally invasive interventional techniques are established treatment methods for PA with favorable success rates and minimal morbidity. The therapeutic options should be tailored to the location, size and rupture risk of PA, condition of the donor artery and existing comorbidity.

Adult ; Aneurysm, False ; diagnosis ; etiology ; therapy ; Embolization, Therapeutic ; Female ; Humans ; Male ; Middle Aged

OBJECTIVETo investigate the effects and mechanisms of Curcumol beta-cyclodextrin Compound (CbetaC) on the proliferation and apoptosis of esophageal carcinoma cell line TE-1.

METHODSThe CbetaC was prepared by saturated solution and confirmed by infrared absorption spectroscopy. The effects of CbetaC (at 25, 50, 100 mg/L) on the proliferation of human esophageal carcinoma cell line TE-1 in vitro was analyzed by MTT assay. The cell cycles and apoptosis were detected by flow cytometer. The relative expression of survivin mRNA was detected by real-time fluorescent quantitative PCR and calculated by the 2(-deltaCt) method. The protein expression of survivin was measured by Western blot.

RESULTSCompared with the control group, results of MTT showed that CbetaC at each dose significantly inhibited the proliferation of TE-1 cells in a dose-dependent manner (P < 0.05). The results of flow cytometry showed that CbetaC resulted in the cell cycle arrest at G0/G1 and G2/M phase, and promoted the cell apoptosis. Besides, when compared with the control group, the protein and mRNA expressions of survivin obviously decreased in each CbetaC group (P < 0.05).

CONCLUSIONSCbetaC could inhibit the proliferation of esophageal carcinoma cell TE-1 and promote the apoptosis. Its inhibition on the survivin expression was correlated with its inhibition on the malignant phenotypes of esophageal carcinoma cells.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Esophageal Neoplasms ; pathology ; Humans ; Sesquiterpenes ; pharmacology ; beta-Cyclodextrins ; pharmacology

OBJECTIVETo evaluate the viral pathogen of pneumonia in children.

METHODSA total of 13 642 cases of children pneumonia in 3 years were enrolled in this study. Antigens of viral pathogen in respiratory excretion, including respiratory syncytial virus (RSV), type 1, 2 and 3 parainfluenza virus, type A and B influenza virus, and adenovirus were detected by direct immunofluorescence method.

RESULTSViral pneumonia accounted for 34.3% of all cases, including 25.8% cases of RSV, 4.7% of parainfluenza virus, 2.4% of type A influenza virus, 0.2% of type B influenza virus and 1.3% of adenovirus. Coinfection was found in 20 cases, in which 17 cases (85%) were infected with RSV and another virus. Positive rates of RSV in children < or = 1 year, 1 to 3 years, and >3 years were 33.1%, 19.7% and 5.1% with a significant difference (chi(2)(trend)=763.4, P < 0.001). The positive rate of adenovirus in children < or =1 year (0.7%) was significantly lower than that in children aged 1 to 3 years and in children >3 years (2.3% and 2.5%) (all P<0.01). The positive rate of type A influenza virus in children aged 1 to 3 years was higher than that in children < or =1 year (chi(2)=18.2, P<0.01). Type 1 parainfluenza virus was found in 1.2% children aged 1 to 3 years with most prevalence (P<0.05). Infection rates of type 3 parainfluenza in children < or =1 year, 1 to 3 years, and >3 years were 4.7%, 3.2% and 1.4% respectively with a significant difference (chi(2)(trend)=52.4, P<0.01). Although there were some differences of infection rate of RSV in different years, it tended to increase from November to next April with a highest rate of 62.8%. Type 3 parainfluenza virus and Type A influenza virus were almost sporadic while type A influenza virus was epidemic in August 2003 with an infection rate of 15.7%.

CONCLUSIONThe highest infection rate of viral pathogen of pneumonia in children is RSV and the follows are parainfluenza, influenza and adenovirus in turn.

Adenoviridae ; isolation & purification ; Adenovirus Infections, Human ; virology ; Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Orthomyxoviridae ; isolation & purification ; Orthomyxoviridae Infections ; virology ; Paramyxoviridae ; isolation & purification ; Paramyxoviridae Infections ; virology ; Pneumonia, Viral ; virology ; Respiratory Syncytial Virus Infections ; virology ; Respiratory Syncytial Viruses ; isolation & purification

To compare the efficacy of various irrigants (citric acid, ethylenediaminetetraacetic acid (EDTA) and NaOCl) and techniques in removing Ca(OH)2 in two types of curved root canal systems, simulated root canals with specific curvatures were used to investigate the effects of different irrigants and instruments on Ca(OH)2 removal. The optimal methods were verified on extracted human teeth. Simulated root canals were assigned to one of two groups based on the irrigation solution:10% citric acid or 2.5%NaOCl. Each group was divided into four subgroups according to the technique used to remove Ca(OH)2. The percentage of Ca(OH)2 removal in different sections of root canals was calculated. On the basis of the results obtained for the simulated canals, 10%citric acid and 17% EDTA were applied to remove Ca(OH)2 from the extracted human teeth with curved root canal systems. The teeth were scanned by micro computed tomography to calculate the percentage of Ca(OH)2 removal in the canals. In simulated root canals, we found that 10% citric acid removed more Ca(OH)2 than 2.5% NaOCl in the 0–1 mm group from the apex level (P<0.05). Ultrasonic and EndoActivator activation significantly removed more Ca(OH)2 than a size 30 K file in the apical third (P<0.05). However, there were no significant differences in any sections of the canals for 10%citric acid or 17%EDTA in removing Ca(OH)2 in extracted human teeth. We concluded that it was effective to remove residual Ca(OH)2 using the decalcifying solution with EndoActivator or Passive Ultrasonic Irrigation in a curved root canal system. A protocol for Ca(OH)2 removal was provided based on the conclusions of this study and the methods recommended in previous studies.