Objective:To study the masseter motor evoked potential(MEP)in patients with sleep bruxism(SB)and in healthy con-trols.Methods:30 subjects with SB and 30 healthy controls were included.MEPs were obtained by transcranial magnetic stimulation (TMS).Tests were done during daytime when the subjects were awake.The data were statistically analysed.Results:In the patients AMT was 55(52,55)%,latency of c-MEP (6.7 ±1.3)ms,the amplitude of c-MEP 0.19(0.15,0.29)mV,latency of r-MEP (2.3 ±0.4)ms,the central conduction time(CCT)4.4(3.3,5.2)ms.In the control subjects AMT was 52(52,55)%,latency of c-MEP (6.4 ±0.7)ms,the amplitude of c-MEP 0.23(0.17,0.28)mV,latency of r-MEP (2.4 ±0.4)ms,CCT 4.0 (3.4,4.4) ms.No significant difference was found between the 2 groups in the measurements evoked by TMS.Conclusion:The MEP after TMS in patients with SB is similar to that of healthy subjects,indicating that the excitability of the cortical motor system is not changed in bruxism subjects,at least when evaluated by TMS.

Objective To evaluate the role of CREB-binding protein (CBP) in cyclooxygenase 2 (COX-2) expression in spinal cord in a rat model of neuropathic pain (NP).Methods Forty male SD rats weighing 220-250 g in which intrathecal catheter was successfully implanted were randomly divided into 4 groups (n=10 each):sham operation group (group Sham),group NP,negative control group (group NC) and group CBP.NP was induced by CCI at 5 days after successful implantation of the intrathecal catheter.Normal saline,negative lentivirus vector (RNAi-NC-LV) and CBP shRNA lentivirus vector (CBP RNAi-LV) 20 μl were injected intrathecally at 7 days after CCI in groups NP,NC and CBP,respectively.Paw withdrawal threshold to mechanical stimuli (MWT)and paw withdrawal latency to thermal stimulus (TWL) were measured at 1 day before operation and at 3,5,7,10,12 and 14 days after operation.The rats were sacrificed after the measurement of MWT and TWL at 14 days after operation and the lumbar segment of the spinal cord was then removed for determination of the expression of CBP,acetylated histone H3/H4 (Ac-H3,Ac-H4) and COX-2 protein (by immunohistochemistry),and CBP and COX-2 mRNA (by RF-PCR).Results Compared with group Sham,MWT and TWL were significantly decreased at different time points after operation in groups NP,NCand CBP,and the expression of CBP,Ac-H3,Ac-H4 and COX-2 protein,and CBP and COX-2 mRNA was up-regulated at 14 days after operation in groups NP and NC (P ＜ 0.05).Compared with group NP,MWT and TWL were significantly increased at 10,12 and 14 days after operation and the expression of CBP,Ac-H3,Ac-H4 and COX-2 protein,and CBP and COX-2 mRNA was downregulated at 14 days after operation in group CBP (P ＜ 0.05).Conclusion CBP is involved in the development of NP by up-regulating COX-2 expression and increasing histone H3 and H4 acetylation in spinal cord.

Adolescent ; Adult ; Burns, Chemical ; Eye Burns ; Fisheries ; Humans ; Hydrogen Sulfide ; Middle Aged ; Occupational Injuries ; Young Adult

Objective To investigate the appropriate setting up of normal reference ranges of lymphocyte subsets in some flow cytometry laboratories and to study the effects of different flow cytometers and various reagents by different manufacturers on the analysis of peripheral blood lymphocyte subsets. Methods Three FCM labs (named A, B and C) in Beijing region were selected representing 3 commonly used flow cytometers (Beckman Coulter Epics XL, Beckman Coulter Cytomics FC500, BD FACS Calibur). 50 samples from healthy donors were distributed to 3 labs and tested according to individual lab's standard operating procedure to verify whether the normal reference ranges of peripheral blood lymphocyte subsets established were appropriate. The application of internal quality control was also investigated. Commercial blood quality control reagents were given to the 3 FCM labs and tested within 20 working days paralleled with routine samples. In addition, 20 patients' samples were prepared using 4 different combinations of reagents ( a , b , c and d). The results from combination a, which used the Beckman Coulter reagents and instrument, were compared to the results from combination b, c and d, which used reagents from different manufacturers. Then the prepared samples were tested on Beckman Coulter Epics XL to evaluate the effects of different combinations of reagents on the results of peripheral blood lymphocyte subsets analyzed by the same instrument. Furthermore, 24 patients' samples prepared by same reagents from Beckman Coulter company were tested on both Beckman Coulter Epics XL and BD FACS Calibur respectively to assess the effects of different instruments on peripheral blood lymphocyte subsets. 20 patients' samples prepared by same reagents and instruments were analyzed by Beckman Coulter Epics XL analytic system and BD FACS Calibur analytic system respectively to assess the effects of the two analytic systems on the lymphocyte subsets. Results Over 10% of the results for NK and T4/T8 in lab A as well as T4 in labs B and C fell outside of their normal reference ranges. The probabilities exceeding corresponding normal reference ranges were 16% ( 9/50 ), 24% ( 12/50 ), 22% (11/50) and 12% ( 6/50 ), respectively. The results using internal blood quality control in 3 FCM labs within 20 working days were all within the reference ranges of the quality control provided by the kit. The biases from b and c reagent combinations were substantial compared with that of reagent a combination. Among the biases from b and c reagent combinations, the lowest probability of bias exceeding 10% was T8 of combination b, which had probability of 70% (14/20). The highest probabilities of hias exceeding 10% were T3 and T4 of b and c reagent combinations, which reached 100% (20/20) . Furthermore, the biases of T3, T8 and B of d reagent combination compared with that of reagent a combination were also substantial. The probabilities of bias exceeding 10% were 35% (7/20) ,85% (17/20) and 75% (15/20), respectively. Comparing the results of samples prepared and analyzed by reagents and instruments from different manufacturers to that of samples prepared and analyzed by the same company's reagents and instruments showed that there were great discrepancies in T3, T4 , T8 , B and NK. The probabilities of bias exceeding 10% were 71% ( 17/24), 80% (19/24) ,38% (9/24), 33% (8/24) and 92% (22/24), respectively. The biases of T8, NK and B were substantial when compared the results from Beckman Coulter Epics XL analytic systems and BD FACS Calibur analytic systems. The probabilities of bias exceeding 10% were 55% (11/20 ), 70% ( 14/20 ) and 55% (11/20), respectively. Conclusions FCM labs should set up their own normal reference range for peripheral blood lymphocyte subsets. The normal reference range should be verified periodically. It is important to apply internal blood quality control regularly and accumulate the quality control results. The reagents and instrument for preparing peripheral blood samples should be from the same manufacturers.

Objective To investigate the correlation between liver-type fatty acid binding protein (L-FABP) and nonalcoholic fatty liver disease (NAFLD) extent and other clinical parameters.Methods Ninety-six patients of NAFLD (NAFLD group) and 100 cases of healthy controls (control group) were selected.The levels of serum L-FABP and blood biochemical parameters were measured by enzyme-linked immunosorbent assay.The lesion degree was assessed by ultrasound.The body mass index (BMI),waist to hip ratio (WHR) and homeostasis model assessment insulin resistance index (HOMA-IR) were calculated.Results The WHR,BMI,fasting plasma glucose (FBG),triglycerides (TG),alanine aminotransferase (ALT),HOMA-IR and L-FABP in NAFLD group were higher than those in control group,there were statistical differences (P ＜ 0.05).The result of correlation analysis showed L-FABP level was positively related with ALT,TG,FBG,WHR,BMI,HOMA-IR (r =0.735,0.728,0.681,0.713,0.699,0.673 ;P ＜0.05),and negative correlation with HDL-C (r =-0.607,P ＜ 0.05).The L-FABP level in control group was (15.42 ± 2.51) g/L,mild NAFLD was (15.96 ± 2.92) g/L,moderate NAFLD was (17.48 ± 3.91) g/L,serious NAFLD was (25.14 ± 5.37) g/L.There was statistical difference in L-FABP level between serious NAFLD and control group,mild NAFLD,moderate NAFLD (P ＜ 0.05).Conclusion Serum L-FABP level of serious NAFLD patient significantly increases,and L-FABP level is related with biochemical parameters of liver function.

Objective　To study the effect of transfecting anti-sense expression vector of the first subtype of the monocarboxylate transporter (MCT1) gene into human lung adenocarcinoma cells on intracellular pH (pHi) regulation, lactate transportation and cell growth. Methods　MCT1 antisense gene recombinant vector pLXSN-MCT1 was introduced into human lung cancer cells A549 with electroporation. The cell colonies resistant to G418 were selected. Positive clones were examined by PCR to confirm the integration of genomic A549 DNA and antisene MCT1 gene. The changes of pHi and lactate transportation were detected with spectrophotometry. Cell growth was studied with cell growth curve. Results　pHi and lactate transport were remarkably decreased in the transfected cells, and the cell growth was inhibited compared with the cells without transfection（P<0.001）. Conclusion　MCT1 gene may play an important role in pHi regulation, lactate transport and cell growth in lung tumor cells.

Objective To investigate the effects of B-type natriuretic peptides (BNP) on expression of monocyte chemoattractant protein-1(MCP-1) in rat peritoneal macrophages and to identify the inflammation-mediated effects of BNP in macrophages. Methods Peritoneal macrophages of primary culture were treated with BNP, BNP+HS-142-1, or BNP+TNF-?+HS-142-1. The protein expression of MCP-1 was measured by Western blot. Results BNP enhanced the MCP-1 protein expression in macrophages, and this effect could be abrogated by HS-142-1. In addition, BNP could inhibit TNF-? induced MCP-1 expression. Conclusion BNP can induce the MCP-1 protein expression in macrophages, suggesting BNP has a pro-inflammatory effect. However, BNP also can inhibit TNF-? induced MCP-1. These findings suggest that the effect of inflammation-mediated by BNP is biphasic though the mechanism is still unclear.

Background The pathogenesis of age-related cataract is associated with the apoptosis of lens epithelial cells (LECs) caused by oxidative stress.Previous studies showed that intracellular focal adhesion kinase (FAK) pathway can be activated by H2O2 in vitro,which induced apoptosis of cells.To investigate the effect of oxidative on FAK expression in LECs is one of important studies in the prevention of age-related cataract.Objective This study was to investigate the expression and function of FAK in human LECs treated by H2O2.Methods Human LECs strain (HLECs-B3) were cultured in vitro in the low glucose DMEM with 10％ fetal bovine serum.Different concentrations (0,30,50,70,100,300,500,700,1000 μmol/L) of H2O2 were added into the culture medium for 24 hours.The survival rate of the cells was detected by Cell Counting Kit-8 (CCK-8) assay.Cell morphology as well as the expression and distribution of FAK in the cells were observed by immunofluorescent staining under the laser confocal microscope.Apoptosis was observed by hoechst33258 staining,and Western blot assay was used to quantitatively detect the expression and phosphorylation of FAK.Results The survival rate of the cells was (1.00±0.03) ％,(1.24±0.03)％,(1.36±0.24) ％,(0.93±0.02)％,(1.75±0.19)％,(1.37±0.18) ％,(0.64±0.01)％,(0.59±0.11)％,(0.14±0.05)％ in 0,30,50,70,100,300,500,700,1000 μmol/L H2O2 groups,with a significant difference among them (F =95.30,P =0.00).The survival rates of the cells in the below 300 μmol/L H2O2 groups were significantly higher than those in the 0 μmol/L H2O2 group,and survival rates of the cells in the above 500 μmol/L H2O2 groups were significantly lower than those in the 0 μmol/L H2O2 group(all at P＜0.05).After H2 O2 treatment for 24 hours,HLECs-B3 cells transformed from polygon shape to spindle shape and extended pseudopodiums,meanwhile the green fluorescence for FAK exhibited in the cytoplasm.Cell apoptosis was found in the 1000 μ mol/L H2O2 group.Western blot assay revealed that the expressing levels (grey scale) were significantly different among the various groups (F=28.08,P=0.00),and FAK expressing levels in the below 300 μmol/L H2O2 groups were significantly higher than those of the 0 μmol/L H2O2 group; while the expressing levels in the above 500 μmol/L H2O2 groups were lower than those of the control 0 μmol/L H202 group (all at P＜0.05).After treated by different concentrations of H2O2,the phosphorylation level of intracellular FAK (p-FAK) was significantly higher in 3 hours group than that in 30 minutes group (all at P＜0.05).Conclusions H2 O2 can affect the survival,proliferation and morphology of human LECs by activating the intracellular FAK pathway,indicating that FAK may play roles in the regulation process of cell biological behavior.

Objective To exPlore the effect of mammalian target of raPamycin ( mTOR ) inhibitor eVerolimus on radiosensitiVity of human non_small cell lung cancer cell line in vitro by using eVerolimus to inhibit mTOR signaling Pathway of A549. Methods Human non_small cell lung cancer cell line A549 was subjected to radiation alone or in combination with eVerolimus treatment. The 50%inhibition concentration ( IC50 ) of eVerolimus in A549 cells was detected by methylthiazol tetrazolium ( MTT) assay in vitro. EVerolimus at the 20%inhibition concentration ( IC20 ) was used to Pretreat A549 cells for 24 h. Cells were then irradiated by X_ray with 2,4,6,8 Gy. The cell surViVal fraction was comPuted by clone formation. Cell surViVal curVe was fitted by multitarget one_hit model, and mean lethal dose ( D0 ), dose quasithreshold ( Dq ), surViVal fraction at 2 Gy ( SF2 ), and sensitization enhancement ratio (SER) were calculated. The exPression ofγ_H2AX was determined by Western blotting and then the relatiVe gray Values were analyzed. Results EVerolimus significantly imProVed the sensitiVity of A549 cells to radiation. The D0 , Dq and SF2 of eVerolimus+irradiation grouP were significantly lower than those of irradiation grouP. The SER was 1. 36. The residual amount of γ_H2AX Protein in the eVerolimus + irradiation grouP was significantly higher than that of the irradiation grouP. Conclusion EVerolimus inhibiting mTOR signaling Pathway can increase the radiosensitiVity of A549 cells.

In order to improve the precision and robustness in determination of soluble solids content ( SSC) of ‘Fuji ’ apple by NIR spectroscopy and eliminate the effect of origin variability on the accuracy of NIR calibration models for the SSC, sample set partitioning based on joint x-y distances ( SPXY) was used to select representative subset from the apple samples of 4 different origins. As a comparison, partial least square ( PLS) was used to establish local origin and hybrid origin models for the prediction of SSC in apple. Competitive adaptive reweighted sampling ( CARS ) and successive projections algorithm ( SPA ) were implemented to select effective variables of the NIR spectroscopy of SSC of apple. The results indicated that the PLS model established based on the 4 origin apple samples performed better than local origin and other hybrid origin models. The model could be effectively simplified using 16 characteristic variables selected by CARS-SPA method from full-spectrum which had 3112 wavelengths. The correlation coefficient (Rp) and root mean square error of prediction (RMSEP) were 0. 978 and 0. 441 oBrix, respectively for SSC. It was found that the model developed by more samples of different origins combined with effective wavelengths showed good prediction ability for apple sample of unknown origin, which indicated that it could significantly reduce the origin effect on the robustness of NIR models for SSC of apple.