Objective To investigate the expression level of nuclear factor-kappa(NF-κB),substance P(SP)and aquaporin 4(AQP4)gene and the change of discharge frequency of pain sensitive neurons (PSN)in the ventral posteriolateral thalamic nucleus(VPL)of acute gout(AG)rats.The neuro-genic inflammation of AG was explored.Methods Forty-eight rats were randomly divided into two groups:the control group and the AG group. According to the time interval after injection of monosodiumurate(MSU)into the the unilateral ankle joint,the AG group was subdivided into seven groups,ie.0.5 hours group,2 hours group,6 hours group,12 hours group,24 hours group,48 hours group and 72 hours group.There were 6 rats in each group.After recording of the discharge frequency of PSN in rats VPL nucleus,the expression level of NF-κB,SP and AQP4 gene in the rats joint synovium were evaluated at mRNA level by PCR in above mentioned time.Results In the 0.5 hours group,the discharge frequency of PSN in the rats VPL nueleus and the expression level of SP gene in the rats joint synovium had increased immediately after the injection of MSU(P＜0.05).In the 6 hours group,they reached the peak level(P＜0.05),and in the 12 hours group,they began to decrease gradually(P＜0.05).In the 0.5 hours group,the expression level of NF-κB and AQP4 gene increased after the injection of MSU(P＜0.05).However,their peak level presented at the 12 hours(P＜0.05),and they decreaged after 24 hours(P＜0.05).The statistical analysis of correlation had shown that there were positive correlation among the expression level of NF-κB,substance P.AQP4 gene and the change of discharge frequency of PSN in the rats VPL nucleus.Conclusion The discharge frequency of PSN in the rats VPL and the expression level of SP gene in the rats ioint synovium can be used to evaluate the Severity of pain in the AG rats.The expression level of NF-κB and SP gene can reflect the severity of neurogenic intlammation.We can know the severity of edema of the joint synovium by detecting the expression level of AQP4 gene.Pain and neurogenic inflammation play an important role in the pathogenesis of AG.Combingelectrophysiology and biochemical technique can shade light on the pathogenesis of AG from different aspects.In the meantime,it may provide a new method for developing new drugs and new approaches for clinical treatment.

OBJECTIVE:To study the effects of 1,25-droxyvitamin D3 on nitric oxide and nitric oxide synthase in peripheral blood mononuclear cells(PBMC)in patients with spinal tuberculosis and to explore its immunoregulation function.METHODS:Patients' PBMC were assigned to test group in which DHVD3 was added or control group in which culture medium was added,then cultured after BCG vaccine inoculation.Cultured supernatant fluid from isolated PBMC was collected at 0,3,6,9,12 days respectively for measuring of relative optical density(OD)of NO by nitric oxide kit.Total RNA extracted from the PBMC was reverse transcribed to cDNA and amplificated by fluorecent real-time quantitative PCR on the sixth day;mRNA of target gene iNOS and reference gene 3-GAPD were analyzed and compared by relative quantitative method 2-??Ct.RESULTS:OD value of NO and mRNA of iNOS in test group were all higher than in control group(P

Objective To investigate the correlation between thrombelastogram (TEG)parameters and plasma D-dimer (D-D) and platelet aggregation rate (PAgT)in patients with acute cerebral infarction (ACI).Methods From January 2015 to Jan-uary 2016,120 patients with ACI (ACI group),and 60 healthy subjects (healthy group)were enrolled in the study.The TEG parameters were detected by thromboelastography,including blood coagulation time (R),blood clotting time (K),rate of blood clotting (αangle),coagulation comprehensive index (CI)and the maximum intensity of blood clotting (MA).Blood coagulation indexes also were detected,including prothrombin time (PT),fibrinogen (FIB,detected by automatic coagulation analyzer),plasma D-D (detected by colloidal gold method)and PAgT (detected by blood aggregation tester).Results R (5.36±0.95 min),K (2.34±0.98 min)and PT (11.88±1.67 s)in the ACI group were shorter than those in the healthy group (6.20±1.28 min,2.87±0.87 min,12.40±1.82 s),(t=4.962,3.547,1.911,all P<0.05).Theαangle (61.65± 7.84)degrees,CI (-0.21±1.11),MA (58.94±6.14 mm),D-D (0.34±0.08 mg/L),PAgT (66.9±6.8)% and FIB (4.41±0.96 g/L)in the ACI group were higher than those in the healthy group [(56.02 ± 6.94)degrees,(-1.50 ± 1.30),(53.86±7.85 mm),(0.24±0.06 mg/L),(55.4±7)%,(2.87±0.88 g/L)],(t=4.714,6.209,5.583,8.550, 10.592,7.039,all P<0.05).In ACI group,D-D,PAgt and the TEG parameters (αangle,CI,MA)were significantly nega-tively correlated (r=-0.481,-0.470,-0.504,-0.488,all P<0.05).D-D,PAgt and the TEG parameters (αangle,CI, MA)were significantly positively correlated (r=0.338,0.395,0.427,0.391,0.436,-0.482,all P<0.05).Conclusion TEG parameters can reflect the severity of ACI to some extent,and they were significantly correlated with plasma D-D and PAgt.

Objective To investigate the effect of hyperuricemia (HUA) on nuclear factor-κB (NF-κB) in myocardial tissue,Mn superoxide dismutases (Mn-SOD) activity in mitochondria,cardiac function of rats with HUA.Methods Sixty SD male rats,including 30 young rats and 30 aged rats,were randomly divided into four groups:the young control group,the young HUA group,the aged control group and the aged HUA group.Fifteen rats were in each group.The HUA rat models were set up by lavage method with yeast extract and ethambutol.Left ventricular systolic pressure (LVSP),left ventricular end-diastolic pressure (LVEDP) and left ventricular pressure at the end of the rising and falling maximum velocity (+dp/dtmax) were determined by intubation in the left ventricle with cardiac catheter.Mn-SOD activity in the mitochondria of rat myocardial tissue was detected by chemical colorimetry.The expressions of NF-κB in rats myocardial tissue were measured by immunohistochemistry (IHC) and reverse transcription polymerase chain reaction (RT-PCR).Differences between groups were analyzed with One way analysis of variance,Student-Newman-Keuls q test was used to compare the mean of multiple samples.Results Hyperuricemia had no significant effect on the cardiac function of HUA groups (P＞0.05).But the gene and protein expression of NF-κB in myocardial tissue and Mn-SOD activity of mitochondria were impacted by HUA.Both the gene and protein expression of NF-κB and the activity of Mn-SOD in HUA groups were increased (F=85.428 4,120.683 0 and 398.228 3,P＜0.01).Furthermore,the increase of gene and protein expression of NF-κB and the Mn-SOD activity in rats myocardial tissue of young HUA group were higher than those in aged HUA group (q=6.818 6,10.693 6 and 18.877 9,P＜0.05).Conclusion Hyperuricemia may cause inflammation and increase oxygen free radicals in myocardial tissue on one hand,and it may reduce inflammation and protect the myocardial tissue by increasing the activity of Mn-SOD in myocardial mitochondria of HUA rats,and removing the oxygen free radicals on the other hand.The response to uric acid in young HUA group is stronger than that in the aged HUA group.

Objective To study the vulnerability of astrocytes bearing mutant SOD1 under the oxidative stress.Methods The cytotoxicity of the serum deprived astrocytes was measured by MTT.The level of ROS was shown by fluorescence of DCF through confocal microscopy.The expression of Nrf2,HO1 and NQO1 in the different cells was detected by Western blot.Results The level of cellular toxicity was higher in the astrocytes bearing mutant SOD1 exposed to the oxidative stress than the astrocytes hearing wild type SOD1.In the astrocytes bearing mutant SOD1,the expression of Nrf2,HO1 and NQO1 decreased.In the presence of mutant SOD1,an unexpected 44 per-cent decrease of Nrf2 was detected.This was associated with a decreases in multiple downstream phase Ⅱ detoxif-ying enzymes and antioxidant enzymes,known as NQO1 and HO1.Furthermore,our results showed that the ex-pression of NQO1 increased 1.5 and 2.5-fold by EGCG at 5 and 10 μmol/L.EGCG also elevated the expression of total Nrf2.Confocal microscopy showed that EGCG caused Nrf2 translocation from the cytoplasm to the nucleus.Conclusion Decrease in Nrf2 expression is the mechanism to explain the vulnerability of astrocytes bearing mutant SOD1 and EGCG strengthened antioxidation function by upregulating the activity of Nrf2.

OBJECTIVE: To establish a method for analysis of proteome of human peripheral leukocytes and to explore the difference between the healthy volunteers and patients with acute lymphocyte leukemia(ALL) in leukocyte proteomics and to study the ALL-related abnormal proteins.METHODS: Peripheral blood samples taken from 10 healthy volunteers and 10 ALL patients were separated for the extraction of leukocyte proteins.The leukocyte proteins were separated by two-dimensional electrophoresis(2-DE).2-DE patterns were analyzed by PDQuest 2D software and the differentially expressed proteins were identified by matrix assisted laser desorption ionization-time of flight-mass spectrometer(MALDITOF-MS) and bioinformatics.RESULTS: The 2-DE patterns of peripheral blood leucocytes from patients with ALL versus those from healthy volunteers were analyzed by PDQuest 2D software and the differentially expressed proteins identified by matrix assisted laser desorption ionization-time of flight-mass spectrometer(MALDI-TOF-MS) were confirmed to beALL-related proteins: AnnexinA5 and cytoskeleton 14 etc.CONCLUSION: Using MALDI-TOF-MS and bioinformatics,the AnnexinA5 and cytoskeleton 14 proteins had been proved to be of close relationship with ALL.The results provided a satisfactory basis for the searching for treatment targets and the molecular targets for the early diagnosis of ALL.

Aim To study effects of metfomin(MF) on calmodulin(CaM) mRNA exprssion of the pitutary-gonad axis in rats.Methods The CaM mRNA expression of the pitutary-gonad axis in rats was measured with the real time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) technique. Results MF (270 mg?kg-1?d-1,ig,21 d) significantly inhibited the CaM mRNA expression in testes (P

Objective To investigate the role of the p38 MAPK pathway in the formation of cytoplasmic vacuoles .Methods Af-ter treated with Anisomycin ,SB203580 or SP600125 ,images of HepG2 ,LM3 ,QBC939 ,Hela and A549 cells were recorded by light microscopy and taken at a magnification of 400 × .The effects of anisomycin ,SB203580 and SP600125 on the activity of p38 and JNK were measured by Western blot .LM3 and A549 cells were stained with the ER-tracker red and the lyso-tracker red and subjec-ted to confocal microscopy analysis .Results (1)Anisomycin could abolish cytoplasmic vacuolization of HepG2 cells .(2)p38 MAPK activation was responsible for anisomycin-induced cytoplasmic vacuolization abolishment .(3)p38 MAPK blocking initiated cytoplas-mic vacuoles formation in various cancer cell lines .(4)p38 MAPK blocking-induced cytoplasmic vacuoles disrupted the integrity of endoplasmic reticulum .(5)p38 MAPK blocking reversibly induced cytoplasmic vacuoles formation .Conclusion These observations provide direct evidence for a role of p38 MAPK signaling in regulating the formation of cytoplasmic vacuoles .

Objective To study the effects of rehabilitation training on the expression of Nogo-A around the area of cerebral infraction using rats. Methods A total of 60 Wistar rats were randomly divided into a reha- bilitation group and a control group after an experimental cerebral infarction had been established in them. The ani- mals in the rehabilitation group were given exercise with a rotating bar, a balance beam and a rolling cage for one hour daily, while those in the control group were caged without any abnormal exercise. Nogo-A expression in the ar- ea surrounding the infarcts was detected by imunohistochemical techniques at the 3rd, 7th, 14th, 21st and 28th day after infarction. Meanwhile, neurobehavioral evaluations were also conducted. Results The animals in the rehabilitation group scored much lower than the controls in the behavioral evaluations at the 14th, 21st and 28th day. The expression of Nogo-A in tissues around the infracted area increased by the 7th day and peaked at the 21st day in both groups, but the expression of Nogo-A was significantly stronger in the rehabilitation group at the 14th, 21st and 28th days. Conclusion Rehabilitation training decreased the expression of Nogo-A in the brain of rats after infarction. This may have important implications for the functional recovery of the central nervous system.

Objective To explore the effect of telomerase reverse transcriptase(TERT) gene transfected bone marrow stem cell(BMSC)on the memory function and hippocampal CA1 region synaptic plasticity in vascular dementia rat.Methods A total of 60 rats were randomly divided into the negative control group(group A),model group(group B),conventional BMSC group(group C) and transfected BMSC group(group D).The related indicators in each group were detected by using the Morris maze test,RTPCR and Western blot respectively.Results The escape latency period in the group C and group D was significantly longer than that in the group B,which in the group D was significantly longer than that in the group C.Compared with the group A,the expressions of brain-derived neurotrophic factor(BDNF)mRNA,TERT mRNA,SYP mRNA and protein in the group B,group C and group D were significantly decreased.The synaptic cleft arrange in group A was clear with more SYN positive ceils.The synaptic cleft in the group D was clearer,and the number of SYN positive cells was close to that in group A.Conclusion TERT transfected BMSC has obvious therapeutic effect on vascular dementia rats and its mechanism may be related to the promotion of BDNF,TrkB expression and the improvement of synaptic plasticity.