Objective To investigate the effects of stress intensity and the expected duration of stress on the inhibition ability of individual responses to stress. Methods A total of 60 cases of hospitalized patients in respiratory department were selected in the study,including 31 male cases and 29 female cases. Incorporated patients were divided into the high-stress group and the low-stress group ( 30 cases in each group) according to whether the patient accepted a invasive examination or not. Then,within each group,pa-tients were further randomly sub-divided into the acute expectation group and the chronic expectation group ( 15 cases in each group) in the form of a lottery. Detection risk disclosure was conducted at 2 hours and at 24 hours before the examination. Visual analogue scale ( VAS) and stop-signal task were used to detect the level of psychological fear and the inhibition ability of individual responses to stress of each group following informing of the detection risk,and the comparative analysis was conducted afterwards. Results ( 1) The score of psychological fear in the high-stress group was significantly increased when compared to the low-stress group ((3.90±2.71) vs (0.80±1.24)),showing statistical difference (F(1,58)=30.16, P<0.01);addi-tionally,there was no statistical difference in the score of psychological fear in subjects between the acute and chronic expectation group ((2.60±2.90) vs (2.10±2.41);F(1,58)=0.785, P>0.05);meanwhile,no statisti-cal difference of the interaction between stress intensity and the expected duration of stress on the level of psychological fear (F(1,58)=0.031, P>0.05). (2) As for stop-signal task,the signal execution error rate of the high-stress group was significantly increased than that in the low-stress group ((9.40±5.80)%vs (8.30± 12.60)%),and the statistical difference was significant (P<0.01).Signal execution responses duration was obviously prolonged in the acute expectation group than that in the chronic expectation group ((677.25±201. 26)ms vs (588.24±127.10)ms),and the difference was statistically significant (P<0.05); meanwhile,stop-signal error rate at 400 ms was significantly decreased ((57.00±26.00)% vs (70.00±23.80)%),and the difference was statistically significant (P<0.05). There was no statistical significance in the interaction be-tween stress intensity and the expected duration of stress (P>0.05) . Conclusion There is no interaction be-tween the effect of the stress intensity and the expected duration of stress on the inhibition ability of individu-al responses to stress. The stress intensity is more important than the expected duration of stress to exert more important influence in the inhibition ability of individual responses to stress.

ObjectiveTo establish the urinary proteome profile of the metabolic syndrome ( MetS ) patients,compare the different urinary proteins between the MetS patients and the normal individuals,and analyze the function of the different proteins,so as to explore the pathogenesis of MetS.MethodsOvernight urine were collected from normal controls (n =6) and MetS patients ( n =6).Acetone precipitation method was used to precipitate proteins of urine.Intra-group proteins were mixed together,identified by reversed phase liquid chromatography-mass spectrometry/mass spectrometry and quantified relatively using spectral counting method.The functions of differential proteins were analyzed using Panther.ResultsA total of 807 and 630 proteins were identified respectively in normal controls and MetS patients.Comparing MetS patients with normal controls,sixty different proteins were found,of which 23 proteins were up-regulated and 37 proteins were down-regulated in MetS patients.In the up-regulated proteins,plasminogen was involved in the plasminogen activation cascade and isoform of alphaenolase,phosphoglycerate kinase 1 and fructose-bisphosphate aldolase B down-regulated in MetS patients were involved in the process of glycolysis and fructose metabolism.ConclusionsThe urinary proteome profile of patients with MetS was established by reversed phase liquid chromatography-mass spectrometry/mass spectrometry.Different proteins between MetS patients and normal people were identified.The plasminogen activation cascade,glycolysis and fructose metabolism play key roles in the pathogenesis of MetS.

(0.05)).There were 5 cases(4.7%) of upper limb edema and functional handicap in PAL group,and 12 cases(11.7%) of upper limb edema and functional handicap in TAL group,there was a statistical difference between PAL group and TAL group(P

Objective To construct fusion gene and prokaryofie expression plasmid encoding the Rv1884c and Rv0867c genes in Mycobacterium tuberculosis (M.tb).The fusion protein wsg expressed efficienfly in E.coli cells.Methods The Rv1884c and Rv0867c genes were amplified by polymerase chain reactions (PCR) with specific primers from genomic DNA of M.tb H37Rv strain,and cloned into pGEX-4Tland pUC19 vectors.Rv1884c and Rv0867c were subcloned into the expression vector pGEX-4T-1 and pPRO-EXHT followed by DNA sequencing.The plasmids were transformed into E.coli DH5α and induced to produce GST-fused Rv1884c and His-fused Rv0867c fusion protein.The protein molecular weight and expression format was analyzed by SDS-PAGE.Results The recombinant expressive vectors pGEX-4T-1Rv1884c and pPRO-EXHT-Rv0867c were constructed.The DH5α strains of E.coli with recombinant plasmid showed high level of Rv1884c and Rv0867c gene expressions after IPTG induction.The SDS-PAGE showed that the plasmids expressed Rv1884c and Rv0867c fusion proteins with molecule weight of 45 000 and 80 000.The recombinant protein accounted for 18.3%and 23.7%of total bacteria protein.The expressed proteins could be purified via GSTrao FF and Ni2+-NTA system kits in denatured condition.Conclusions Mycobacterium tuberculosis Rv1884e and Rv0867c genes have been cloned and expressed Successfully in E.coli DH5α.The results lay a basis for further study of fast culfivation in Mycobacterium tuberculosis and investigation of their activities and functions.

Objective To investigate the expression of protein interacting with Cα kinase 1 (PICK1) and its significance in colorectal adenocarcinoma.Methods The expression of PICK1 were detected by immunohistochemistry and Western blot methods in tissues of colorectal adenocarcinoma,colorectal adenoma,colorectal polyp and adjacent tissues.The correlation between PICK1 and clinical pathological data was explored.Results Immunohistochemical assay showed that the positive ratio of PICK1 protein was 72.5 ％ (74/102),and overexpressed in 31 cases (30.4 ％,31/102) with colorectal adenocarcinoma.The ratio of high expression level of PICK1 in colorectal adenoma tissues (22.2 ％,8/36) was significantly higher than that in the adjacent tissues (0,0/40) (P ＜ 0.05).The ratio of high expression level of PICK1 in colorectal polyp tissues (5.6 ％,2/36) was no statistically difference compared with that of the adjacent tissues (P ＞ 0.05).Western blot analysis revealed that the expression of PICK1 in colorectal adenocarcinoma (1.10±0.04) was significantly higher than that in the adjacent tissues (0.75±0.07) (P ＜ 0.05).The result showed significant correlation with the tumor location,the degree of differentiation,depth of invasion,TNM stages and lymph metastasis (all P ＜ 0.05).However,there is no correlation was revealed between PICK1 expression and the patients age,gender (both P ＞ 0.05).Conclusion The expression of PICK1 may be associated with the tumorgenesis,progression,invasion and metastasis of colorectal adenocarcinoma,which contributes to the prognosis of patients.

Objective To evaluate the effects of penehyclidine hydroehloride as an atropine alternative on angioearpy and glandular secretions when premedieated in ketamine complex total intravenous anesthesia(TIVA)in children.Methods Forty patients aged 3-10 years undergoing ketamine and propofol complex TIVA were randomly divided into two groups.Penehyclidine hydrochloride(group P,n=20)or atropine(group A,n=20)was premedicated intramuscularly 30 min before anesthesia.Heart rate(HR),mean arterial pressure(MAP),breath rate(R)and the amount of saliva secretion(SS)were recorded before premedication(0 min),10 min,20 min,30 min,60 min and 150 min after.Results (1)SS reduced significantly 20 min,30 min and 60 min after premedication in both groups(P＜0.01),and in 150 min,it was still in a significantly reduced level in group P(P＜0.01),which was significantly lower than that in group A(P＜0.01).(2)MAP,HR and R in group P showed no significant differences before and after premedication(P＞0.05).But in group A,HR increased significantly at 20 min,30 min and 60 min after premedication(P＜0.05 or＜0.01),MAP increased significantly at 30 min and 60 min after premedication(P＜0.01),and meanwhile of them were also significantly higher than those in group P(P＜0.05 or＜0.01).Conclusions Penehychdine hydrochloride can effectively reduce respiratory glandular secretion with longer persistence,and nearly has no influence on HR and blood pressure,which suggests it could be a superior to atropine alternative as an anesthesia premedication in children.

Objective To explore the effects of HINT1 on the proliferation and apoptosis of human melanoma cell line A375.Methods Three cell clones were used in the experiment.including A375 cells that were previously transfected with eukaryotic expressing vector pcDNA.3.1/myc-His(-)A-HINT1 and highly expressed HINTI(HINT1-A375),A375 cells transfected with pcDNA.3.1/myc-His(-)A empty vector(neo-A375)and untransfected A375 cells.M3T assay,flow cytometry and terminal deoxynucleotjdyl transferase (TdT)-mediated dUTP nick end labeling(TUNEL)were performed to detect the proliferation,cell cycle and apoptosis of the three kinds of cells,respectively.The relative activity of Caspase 3,8 and 9 was measured by spectrophotometry,and the protein expression of Bcl-2,Bax,Cytoehrome C and p53 by Western blot. Results Compared with neo-A375 and untransfected A375 cells,HINT1-A375 cells grew more slowly(P<0.05)with an increase in G1-phase population(73.17%±3.99%,F=25.65,P<0.05).a decrease in S-phase population(16.75%±1.62%,F=75.48,P<0.01),and a pronounced late apoptosis(23.57%±9.58%,F=11.71,P<0.01).A significant increase was also observed in the percentage of apoptotic cells(12%±1%,F=358.02,P<0.01)as well as in the relative activity of Caspase 9(0.45±0.03,F=135.62,P<0.01)and Caspase 3(0.46±0.04,F=90.28,P<0.01)in HINT1-A375 cells compared with the other two kinds of cells.Western blot showed upregulated expressions of Bax,Cytochrome C and p53 but downregulated expres-sion of Bel-2 in HINT1-A375 cells.Conclusions The overexpression of HINT1 could inhibit the proliferation and promote the apoptosis of A375 cells with cell cycle arrest in G1 phase,hinting that HINT1 may be a tumor suppressor gene in human melanoma.

Objective To study the effects of imatinib mesylate on the apoptosis in human melanoma cell line M14.Methods M14 cells were cultured in vitro in the presence of imatinib mesylate at three concentrations(5,10 and 20 μmol/L)for 96 hours.Sebsequeutly,annexin V-FITC and propidium iodide(PI)double staining flow cytometry and terminal deoxvnucleotidvl transferase (TdT)-mediated dUTP nick end labeling (TUNEL)were used to detect the cell cycle and apoptosis,respectively,DAPI staining to observe the mot-phological changes.and Western blot to measure the protein expressions of bcl-2 and bax in cells.Results Imatinib mesylate of the three concentrations could induce an evident increase in the apoptosis in M14 cells.Compared with untreated M14 cells,an increase of cell population in S phase was observed in imatinib mesylate.treated cells(P<0.05),along with a decline in cell population in G2/M phases(P<0.01).Annexin V/PI double staining and TUNEL revealed a significant increase in the rate of early apoptosis and in the acount of apoptotic cells,respectively,in M14 cells treated with imatinib mesylate of the three concentrations(all P<0.01).After treated with imatinib mesylate of 20 μmol/L.there was a morphological change characteristic of apoptosis in M14 cells,together with an upregulated expression of bcl-2(t=15.46,P<0.01)and downregu-lated expression of bax(t=25.53,P<0.01).Conclusions Imatinib mesylate can interfere with the process of cell cycle of and induce the apoptosis in M14 cells,which may be mediated through mitochondrial pathway.

Objective To improve the understanding of cutaneous Mycobacterium haemophilum infection.Methods Clinical and laboratory data were collected from two patients with cutaneous Mycobacterium haemophilum infection.The clinical and histopathologic manifestations, etiology and treatment of cutaneous Mycobacterium haemophilum infection were analyzed.Results The two patients were both immunocompromised.Lesions, which occurred after trauma in both the patients, began as well-marginated subcutaneous nodules, and then gradually progressed into ulceration with crusting and abscess formation.Histopathological examination revealed epithelial cell granuloma or histiocytic infiltrate in the dermis, as well as focal necrosis and neutrophil abscesses.Acid-fast staining showed abundant bacilli in tissues.These bacilli only grew in iron-containing medium, and were finally identified as Mycobacterium haemophilum by PCR and sequencing.According to the result of antimicrobial sensitivity testing, both the patients were treated with rifampicin, clarithromycin and moxifloxacin.Three months later, the condition was improved in both of them.Conclusions Mycobacterium haemophilum infection occurs most frequently in immunocompromised populations after trauma, manifests as subcutaneous nodules and abscesses with histopathological changes consistent with infectious granuloma.Molecular biological approaches are reliable for the identification of Mycobacterium haemophilum.

Objective:To explore the expression of peptidyl prolyl cis-trans isomerase (Pin1) activity in peripheral blood of patients with rheumatoid arthritis (RA) and the value and correlation of T helper cell 17/regulatory cells (Th17/Treg) cells, and to analyze the effect and influence of all-transretinoic acid (ATRA) on it.Methods:① Comparing the difference of Pin1 expression and absolute counts of Th17 and Treg between RA patients before and after treatment and healthy control group, Kruskal-Wallis rank sum test was used for analysis. ② To analyze the correlation between the expression of Pin1 and its general data, activity indicators [such as erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and disease activity score 28 joints (DAS28) scores], Th17, Treg and some cytokines in RA patients, and to use Pearson and Spearman correlation tests. ③ To analyze the difference of Pin1 expression and Th17/Treg in peripheral blood of RA patients treated with low-dose all-trans retinoic acid (10 mg twice a week) and traditional immunosuppressants such as hydroxychloroquine for 3 months respectively. Mann-Whitney U test was used for comparison between the two groups, and the difference was statistically significant with ( P<0.05). Results:① The activity of Pin1 in peripheral blood of the newly treated group of RA was [13.62(9.16, 19.42)] higher than that of the healthy control group [8.97(7.62, 11.45)]( Z=42.82 , P<0.05), and Th17 was [18.28(12.76, 24.08)] higher than that of the healthy control group [6.04(4.96, 4.96)]( Z=48.83 , P<0.05). Treg [11.06(5.31, 21.87). It was lower than that of healthy control group [40.41(24.33, 48.52)]( Z=42.21 , P<0.05). ② the activity of Pin1 in peripheral blood of RA patients was positively correlated with CRP, the number of involved joints, DAS28 score, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) ( r=0.396, P<0.05; r=0.683, P<0.05; r=0.466, P<0.05; r=0.315, P<0.05; r=0.416, P<0.05). ③ Compared with the newly treated RA group, the activity of Pin1 [6.94(5.96, 8.77), Z=42.82 , P<0.05] and Th17 7.38 decreased [7.38(3.85, 11.21), Z=48.83 , P<0.05], while Treg [40.41 (17.77, 33.47)] increased ( Z=42.21 , P<0.05). ④ Compared with the traditional medicine group, Treg [28.9(21.73, 37.36)] was higher in the retinoic acid group, and the difference was statistically significant ( Z=-2.683 , P<0.05). The activity of Pin1 was [6.23(5.58, 8.75)], but there was no statistical significance ( Z=-1.622 , P=0.104). Conclusion:Pin1 in peripheral blood of RA patients is over-expressed. Th17 is increased and Treg is decreased. ATRA combined with other traditional drugs can reduce Pin1 activity, promote Treg growth and improve disease activity of RApatients to a certain extent.