Objective To study calcium chelator BAPTA-AM antagonize the cellular oxidative stress induced by hydrogen peroxide ( H2 O2 ) and to explore the effect of calcium ion on the cell degeneration mediated by NLRP3.Methods The SHSY5Y cell model of oxidative stress was made by hydrogen perox-ide,then the cell model was treated with calcium ion carrier A23187 or BAPTA-AM,a higher efficiency cal-cium chelating agent.The cells were divided into 4 groups:H2O2 treatment group,H2O2+A23187 group, H2 O2+A23187 +BAPTA-AM group and control group.NLRP3 protein was detected by Western blot,and Caspase-1 and IL-1βwere detected by ELISA.Results NLRP3 expression was significantly increased in cells treated by hydrogen peroxide(P<0.05) .The NLRP3 protein continued to increase, and the expression of Caspase-1((57.1±19.2)pmol/L) and IL-1β((484.2±49.5)pg/ml) protein was also increased signifi-cantly in cells treated by A23187,and the difference had statistically significant for caspase-1 or IL-1βin H2O2+A23187 group compared with those in control group(Caspase-1:(26.8±12.9)pmol/L,IL-1β:(326.9 ±52.1) pg/ml) (P<0.05, P<0.01) .NLRP3,Caspase-1 and IL-1βwere all significantly reduced after adding a high-er efficiency calcium chelating agent BAPTA-AM.Conclusion Calcium overload is likely to enhance the oxidative stress induced by hydrogen peroxide and engender neurodegeneration mediated by NLRP3 inflammasome.

Phytoene synthase is a rate-limiting enzyme in the pathway of carotenoid biosynthesis.To aim at establishing a transformation of Ginseng callus cells, elevating the nutritive value through encouraging the composition of corresponding carotenoid,taking Ginseng callus cells as acceptor,psy gene were transformed into cells via Agrobacterium-mediated. The factors affecting genetict ransformation were also investigated seperately from concentrations of A.tumefaciens, age of Ginseng callus cells, infection time and co-culture time. PCR,PCR-Southern and RT-PCR analysis from the transferred gene plants proved that psy gene has been transferred into Ginseng callus cells and can be expressed. The content of ?-carotene was increased by an average of 26 times.A base of improving and raising the contents of carotenoid in the Ginseng callus cell was established.

Objective To explore the role of nucleotide-binding oligomerization domain receptor protein 3 (NLRP3) inflammasome in the pathogenesis of dementia. Methods 16 patients with with Alzheimer's disease (AD), 22 patients with vascular dementia (VD) and 20 healthy persons with similar age were detected the level of NLRP3 mRNA with RT-PCR in peripheral blood mononuclear cells January, 2014 to October, 2015. The serum level of interleukin (IL)-1βand IL-18 were detected with ELISA. The total number of white blood cells (WBC) and serum total calcium were collected. Single factor correlation analysis was performed between NLRP3mRNA and IL-1β, or IL-18, or WBC, or calcium concentration. Results The levels of NLRP3 mRNA ranged from high to low were as in AD group>VD group>the healthy group (q>11.48, P<0.05). The serum level of IL-1βwas higher in AD group than in the the healthy group (q=16.74, P<0.05), but not significantly different between VD group and the healthy group, nor between VD group and AD group (P<0.05). There was no signifi-cant difference about IL-18 among AD group, VD group and the healthy group (P<0.05). NLRP3 mRNA was positively correlated with the serum levels of IL-1β(r=0.64) and calcium (r=0.58) in AD group. There was no correlation between NLRP3 mRNA and IL-18 or WBC (P<0.05). Conclusion The expression of NLRP3 inflammasome may be related with the pathogenesis of AD, but little impact on the pathogene-sis of VD.

Several specific monoclonal antibodies for B, T and natural killer (NK) cell were used to investigate the B cell localization and the expression of their phe- notype in lymphoid nodules on frozen and paraffin sections of human tonsil and lymph node by means of an immunocytochemical ABC technic. The results indi- cate that monoclonal antibodies reactive with germinal centers in tonsil and ly- mph node gave a simlar results and the results indicate that transformation and germination of germinal center cells involve phenotype changs but except T-200. For example, in the lower zone of germinal center, the lymphoblasts are weakly stained for IgM andLN-2 antibodies, but not for OKB-2 and BA-1, while in th upper zone the centrocytes are intense staining for IgM, LN-2, OKB-2 and BA-1 antibodies arelight or moderate staining separately Further charaterization of B cells in upperzone is frenquently observed clcavages on their nuclear memb- rane. In the mantle zone, the lymphocytes are strongly reacted with OKB-2 and BA-1, middle staining for LN-2 and light staining for sIgM. Plasma cell is only reactive with T-200 and IgM antibodies.

Background The cryptochrom 2 (Cry2)in mammalian retina is a main influential factor of circadian clock.Objective Purpose of this study was to investigate the effect of light exposure rhythm on expression of Cry2 in retina.Methods Thirty clean healthy Sprague Dawley(SD)rats were collected and divided into two groups randomly.The rats of the control group exposed to natural light with the normal rhythm for 30 days,but rats of the experimental group exposed to the artificial light (light: dark =18 hours:6 hours) for 3 months with the light intensity of(533± 16)lx.The histopathological change and ultrastructural alteration of rat retina in both groups were examined under the light microscope and transmission electron microscope at the end of the experiment.Expressions of Cry2 protein and its mRNA were assayed by immunohistochemistry and quantitative PCR(Q-PCR).Results The rat retinal morphology and ultrastructure were clear and order-arranged under the light microscope and transmission electron microscope in the control group.However,atrophy and disorganization of retina were found under the light microscope,and liquefaction and vacuole of outer segments of photoreceptors were observed.The vacuolar degeneration of mitochondria in the inner segments of photoreceptors,cellular nuclear shrinkage,chromatin margination,nuclear notch and destruction were seen in the outer nuclear layer under the transmission electron microscope.Immunohistochemistry showed that the expression of Cry2 protein located in cytoplasm and nuclei membrane of the retinal ganglion cell layer and inner nuclear layer in both normal rats and experimental rats.The scores of Cry2 protein expression were 0.833±0.197 in the experimental group,and 1.700±0.245 in the control group,with a significant difference between them (P=0.009).The quantities of Cry2 mRNA were 0.962 ± 0.125 in the control group and 0.615±0.100 in the experimental group,showing a significant difference between the two groups (P =0.006).Conclusions Long-term light exposure under the 533 lx leads to retinal structural and functional damage probably by down-regulating Cry2 expression in retina.Whether the regulation of Cry2 expression is helpful for stabilizing the biorhythm or not is a worthy question to explore.

Objective To optimize and establish ELISpot assay for CCP/AST which could secrete IFN-γ and IL-4, and explore the role and clinical significance of CCP/AST cells in occurrence and development of RA disease. Methods CCP was used as specific-stimulator with FLAG peptide as a control,the frequencies of positive SFC which could specifically secrete IFN-γ and IL-4 in 64 cases of RA, 64 cases of non-RA autoinunune diseases and 30 cases of healthy individuals were tested by ELISpot technique. The diagnostic value of CCP/AST cells was evaluated in patients with RA disease. Meanwhile, the relationships among the indexes above and patient joint symptoms as well as other laboratory parameters were further analyzed and discussed. Results The results showed that the mediam numbers of IFN-γ-SFC and IL-4-SFC were 39(12-77)/3 x 105 PBMC and 1 (1-3)/3 × 105 PBMC in RA patients, the positive rates were 81.3% and 18. 8% respectively. The median value of IFN-γ-SFC/IL-4-SFC ratio was of 15(5-39), the positive rate was of 78. 1%. Both IFN-γ-SFC and ratio of IFN-γ-SFC/IL-4-SFC were significantly higher than those of non-RA diseases (Z = - 7. 458, - 7. 019, P < 0. 01 ) and healthy control ( Z = - 6. 643, - 5. 760, P <0. 01 ), also both these parameters in RA patients with positive anti-CCP antibody and negative anti-CCP antibody were significantly higher than those patients with systemic lupus erythematosns ( Z = - 6. 573, - 6. 098, - 4. 552, - 4. 726, P < 0. 01 ), ankylosing spondylitis ( Z = - 3. 520, - 3. 326, - 2. 950,-2. 126, P<0. 01 or 0. 05), other autoimmune diseases (Z = -4. 838, -4. 418, - 3. 681, -3. 839,P < 0. 01 ) and healthy controls ( Z = - 6. 553, - 5. 578, - 4. 635, - 4. 163, P < 0. 01 ). Combining IFN-γ-SFC, IL-4-SFC with IFN-γ-SFC/IL-4-SFC for RA diagnosis, the area under curve of receiver operating characteristic( ROCAUC) and Youden index were 0. 910 and 0. 747. The diagnostic sensitivity and specificity were 87. 5% and 87.2%. Positive and negative predictive values were 82. 4% and 91.1%, respectively. Correlation analysis showed that ratio of IFN-γ-SFC/IL-4-SFC was not only significantly correlated with antiCCP antibody(r =0.393, P <0.01), but also correlated with joint symptoms in patients such as with number of joints swelling-pain ( r = 0. 429 , P < 0. 01 ), number of joints damage ( r = 0. 463, P < 0. 01 ),rheumatoid factor (r = 0. 166, P < 0.01) and erythrocyte sedimentation rate (r=0. 199,P<0.05).Conclusions There is widely existence of CCP/AST cells activation and abnormity in RA patients,indicating higher frequency of CCP-specific Th1 cells. These results provides a new experimental evidence for an objective understanding the function of specific cellular immune responses and cytokine network regulation mediated by citrullinated proteins in RA. So, the assay for CCP/AST cells has potential values in RA diagnosis and clinical application.

OBJECTIVETo analyze the relation of plasma D-dimer levels and incidence of deep venous thrombosis after spinal surgery.

METHODSThe clinical data of 63 patients underwent spinal surgery from October 2009 to October 2010 were retrospective analyzed. There were 40 males and 23 females with an average age of 48 years old(21 to 76) in operation. Operation levels of 15 cases were in cervical vertebrae, 4 cases were in thoracic vertebrae,and 44 cases were in lumbar vertebrae. Thirty patients with spinal fracture were caused by trauma and 33 patients without trauma, 11 patients combined with nerve injury. The patients were divided into two groups according to plasma D-dimer levels, more than or equal to 500 microg/L was D-dimer positive group and less than 500 microg/L was D-dimer negative group. Venous blood of all patients early morning with empty stomach were testd on admission, and at 2 h, 1 d, 2 d, 3 d, 4 d, 6 d, 8 d, 10 d, 15 d after operation,respectively.

RESULTSThere was no statistically significant differences in sex, operative segments, implants, operative posture, age, bleed volume, body weight, peroperative D-dimer levels between two groups. After operation, plasma D-dimer of 19 patients were more than or equal to 500 microg/L, with persistent or progressive increasing. Two cases occurred deep venous thrombosis in D-dimer positive group, they respectively were found at 3 days and 8 days after operation. Both of them underwent posterior decompression and internal fixation. However,no deep venous thrombosis was found in D-dimer negative group.

CONCLUSIONPostoperative D-dimer assay can effective predict deep venous thrombosis occurrence. D-dimer level more than or equal to 500 microg/L will be considered as a risk factor for deep venous thrombosis after spinal surgery.

Adult ; Aged ; Female ; Fibrin Fibrinogen Degradation Products ; metabolism ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Spine ; surgery ; Ultrasonography ; Venous Thrombosis ; blood ; diagnostic imaging ; surgery ; Young Adult

AIM: To assess the relative agreement of GAT and NCT in IOP measurement by comparing the differences between Goldmann applanation tonometer (GAT) and non-contact tonometer (NCT) in intraocular pressure (IOP) detection.METHODS: IOP of 529 eyes of 265 volunteers were measured with both NCT and GAT, respectively.RESULTS: The measurement results of NCT were lower than that of GAT, there was significant difference between the IOP measured with NCT and GAT (19.13 vs23.43, t=22.644, P＜0.05). With the increasing of IOP values, the difference magnitude was greater, especially in IOP group that was more than 30mmHg, but the correlation coefficient became lower.CONCLUSION: The measurement results with NCT are lower than that of GAT. When the IOP with the NCT is in borderline value, it need be corrected with GAT, in order to discover the pathologically elevated IOP and avoid the misdiagnosis and mistreatment of glaucoma.

Objective To discuss the feasibility of preoperative diet by measuring gastric emptying time of carbohydrate and protein nutrient solutions in healthy volunteers.Methods A total of 20 healthy volunteers were collected from August 2013 to May 2014.On the morning of the trial,baseline gastric residual volume of each volunteer was measured with magnetic resonance imaging at 8 a.m.,then each of the 20 healthy volunteers took 12.5％ carbohydrate solution 400 ml (containing 40 g of maltodextrin and 10 g of sucrose) or 12.5％ whey protein solution (containing 50 g whey protein) in 5 minutes.Magnetic resonance imaging was conducted to measure the gastric residual volume every 25 minutes.The volunteers were shifted to the other nutrient solution after a 1-week interval.The gastric emptying time of both nutrient solutions was calculated to generate the curves illustrating the process of gastric emptying.Results The baseline gastric residual volume of the volunteers was (14.90 ± 9.39) ml.The total gastric emptying time of carbohydrate solution was (104.90 ± 27.98) min (95 ％ CI 98.64-111.16 min),while that of whey protein solution was (199.6 ± 34.17) min (95％ CI 184.47-214.73 min).There was a significant difference between these two types of nutrient solution in terms of gastric emptying time (P ＜ 0.000 1).Conclusions The induction of anesthesia could be performed 2 hours after carbohydrate administration,and at least 4 hours after whey protein administration.

Objective To determine the gastric emptying time for liquids in the healthy volunteers by magnetic resonance imaging (MRI) and to provide a reference for reasonable preoperative fasting time.Methods Nineteen healthy volunteers of both sexes,of ASA physical status Ⅰ or Ⅱ,aged 20-60 yr,were enrolled in the study.The volunteers were fasted from the intake of liquids or solids starting from 22:00 the night before the trial,and 12.5％ carbohydrate solution 400 ml containing 40 g maltodextrin and 10 g sucrose was given orally.MRI was performed to measure the baseline gastric fluid volume at 8:00 on the day of the trial (T0).The gastric fluid volume was measured immediately after administration of the oral solution,and then measured every 25 min until the gastric fluid volume was returned to the baseline before administration of the oral solution or to ＜25 ml,and was recorded as T25,T50,T75,T100,et al.The gastric fluid volume was drawn using a computer to obtain the curve for gastric emptying.The gastric half and total emptying time was calculated using the curves.Results The gastric half emptying time was (32± 12) min,and the gastric total emptying time was (99±22) min in the volunteers.Compared with those at Tb,the gastric fluid volume was significantly increased at T25,T50,T75,T100,and no significant change was found in gastric fluid volume at T125,T150and T175.Conclusion After oral intake of liquids,the gastric emptying time is about 2 h,indicating that the preoperative fasting time for liquids can be shortened to 2 h before anesthesia in the healthy volunteers.