Objective To evaluate the changes in the expression of NRF-1 in the spinal cord during remifentail-induced hyperalgesia in a rat model of incisional pain.Methods Forty-eight Sprague-Dawley rats,weighing 180-220 g,were randomly divided into 4 groups (n =12 each) using a random number table:control group (C); incisional pain group (group Ⅰ); remifentanil group (group R); incisional pain + remifentanil group (group Ⅰ + R).All the rats were anesthetized with sevoflurane.A 1-cm longitudinal incision was made through skin,fascia and muscle of the plantar aspect of the right hindpaw in I and I + R groups.In C and I groups,normal saline was subcutaneously infused for 30 min.In group I + R,remifentanil (0.04 mg/kg,0.4ml) was subcutaneously infused for 30 min starting from the onset of skin incision.Paw withdrawal threshold to mechanical stimulation (PWMT) was measured at 24 h before operation and at 2,6,12,24 and 48 h after operation.After measurement of PWMT at 48 h,the rats were sacrificed and L4,5 segments of the spinal cord were removed rapidly to detect the expression of nuclear respiratory factor 1 (NRF-1) by immunofluorescence and Western blot.Results Compared with group C,PWMT was significantly decreased at each time point after operation,and the expression of NRF-1 in the spinal cord was up-regulated in I and I + R groups (P ＜ 0.05).Compared with group I,PWMT was significantly decreased at each time point after operation,and the expression of NRF-1 in the spinal cord was up-regulated in group I + R (P ＜ 0.05).Conclusion Up-regulation of NRF-1 expression in the spinal cord may be involved in the development of remifentail-induced hyperalgesia in a rat model of incisional pain.

Objective To find out more about the population structure and clonal complex of clinical Vibrio parahaemolyticus strains in Asia.Methods Clinical Vibrio parahaemolyticus strains data were screened in Asia with complete ST and pST types from PubMLST public database,their subgroup and complex were analyzed,and the minimum spanning tree based on ST and pST types respectively was completed.Results From the database,341 items of ST and pST types of Asian clinical Vibrio parahaemolyticus strains were screened,including 157 ST,most of which were of ST3 type.Totally 214 items of data came from China (including Hong Kong and Taiwan),and covered 133 ST,most of which were of ST3 type.eBURST software was used and 17 groups and 94 singletons were found.Software STRUCTURE analysis showed that the appropriate subset number of clinical V.parahaemolyticus strains in Asia was 7,and that the average distance between samples in each subgroup was 0.9113.Conclusion Clinical V.parahaemolyticus strains in Asia show high diversity and can be subdivided into seven subgroups.ST3 type is dominating when multilocus sequence typing(MLST)is used and pST2 type is the majority by AA-MLST typing.

Objective To learn more about virulence genes and clonal complex group structure of Vibrio parahaemolyticus (VPH)strains separated from aquatic products in East China coastal areas between 2007 and 2012.Methods Seventy-nine strains separated from aquatic products in eastern coastal areas(Guandong,Fujian,Shanghai,Shandong,Jiangsu and Beijing)of China between 2007 and 2012 were identified as VPH by real-time-PCR with gene tlh.Gene tdh,trh and orf8 were also detected.Subgroup analysis and complex analysis were conducted of the VPH strains to build the minimum spanning tree respectively based on ST and pST types.Results In 79 VPH strains,gene tlh was positive while 8.86%(7 /79)of the isolates of gene tdh were positive.The carrying rate of gene orf8 was 8.86%.These 79 strains were of 69 ST types,involving 3 clonal complex and 62 singleton.By amino acid(AA)-multilocus sequence typing(MLST),79 strains covered the 23 pST types,2 clonal complexes and 1 singleton.The 363 single nucleotide polymorphisms(SNPs)were divided into 68 patterns.Conclusion VPH strains from aquatic products in eastern coastal areas of China are characterized by high polymorphisms and a low carrying rate of virulence genes.ST3 type is dominating when MLST typing is used while pST1 type is the majority by AA-MLST typing.

Objective To evaluate the changes in the expression of Toll-like receptor 4 (TLR4) mRNA and its down-stream cytokines IL-1β and TNF-α mRNA in spinal cord in a rat model of incisional pain.Methods Fifty-eight male Sprague-Dawley rats,weighing 180-220 g,were used in this study.A 1-cm longitudinal incision was made through skin,fascia and muscle of the plantar aspect of the hindpaw in isoflurane-anesthetized rats.Mechanical paw withdrawal threshold to yon Frey filament stimulation (MWT) on the operated and non-operated sides was measured before operation and at 0.5,1,2,6 and 12 h and 1,2,3,5 and 7 days after operation.Six rats were chosen and sacrificed before operation and at 2 and 8 h and 1,2,3,5 and 7 days after operation.Their lumbar segments (L4-6) of the spinal cord were removed for determination of the expression of TLR4,IL-1β and TNF-α mRNA by real-time PCR.Results Compared with the baseline value before operation,MWT on the operated side was significantly decreased at 0.5 h-5 days after operation,and the expression of TLR4,IL-1β and TNFα mRNA was up-regulated at 2 and 8 h and 1,2 and 3 days after operation (P ＜ 0.05),and no significant change was found in MWT on the non-operated side (P ＞ 0.05).MWT on the operated side was lowest at 2 h after operation and then gradually increased,the expression of TLR4 mRNA peaked on 1 day after operation,and the expression of IL-1 and TNF-α mRNA peaked at 8 h after operation (P ＜ 0.05).The TLR4 mRNA expression was negatively correlated with MWT on the operative side (r =-0.484,P ＜ 0.05),and IL-1 mRNA and TNF-α mRNA expression was positively correlated with TLR4 mRNA (r =0.294 and 0.540,respectively,P ＜ 0.05).Conclusion The expression of TLR4 mRNA and its down-stream cytokines IL-1β and TNF-α(mRNA in spinal cord is up-regulated,this change is involved in the maintenace of incisional pain,but it does not play an important role.

Objective To investigate the effect of diabetes on the expression of the glucose transporter (GLUT-1) in blood brain barrier (BBB) endothelial cell at different time points. Methods Forty male SD rats were randomly divided into normal control and diabetic group. Diabetic model of SD rats was established by intravenous streptozotocin(50 mg/kg) injection. Ten rats in each group were sacrificed on the 2nd month and 10th month respectively. The rat' s prefrontal cortex was removed to observe the expression of GLUT-1 in BBB EC with immunohistochemical(IHC) staining. Results By IHC method, the number of GLUT-1 positive microvessels was significantly decreased in diabetic group in the 2nd month (4. 3 ± 0. 9) compared with that of control group (6.4 ± 1.7, t = 7. 586, P < 0. 01 ) ; the number of GLUT-1 positive microvessels was significantly increased in the diabetic group in the 10th month (8.4 ± 1.4) compared with that of control group (5. 8 ± 1.7, t = 15.922,P < 0. 01 ). Conclusions Short-term diabetes GLUT-1 expression in BBB EC is downregulated in early stage, while it is upregulated in advanced stage.

A rapid, sensitive and convenient LC-MS/MS method was developed for the determination of γ-aminobutyric acid (GABA) in human plasma. d2-γ-Aminobutyric acid (d2-GABA) was synthesized as internal standard (IS). After extraction from human plasma by protein precipitation with acetonitrile, all analytes were separated on a Luna HILIC column (100 mm x 3.0 mm, 3 μm) using an isocratic mobile phase of water: acetonitrile: formic acid (20 : 80 : 0.12) with a flow rate of 0.5 mL x min(-1). Acquisition of mass spectrometric data was performed in multiple reaction monitoring mode (MRM) in positive electrospray ionization using the transitions of m/z 104 --> 69 for GABA and m/z 106 --> 71 for d2-GABA. The method was linear in the concentration range of 5.00 to 1 000 ng x mL(-1). The intra- and inter-day precisions were within 9.9%, and accuracy ranged from 99.1% to 104%, within the acceptable limit across all concentrations. The method was successfully applied to a pharmacokinetic study of GABA tablets in healthy Chinese volunteers.
Chromatography, Liquid ; Humans ; Tandem Mass Spectrometry ; gamma-Aminobutyric Acid ; blood

Objective To survey the prevalence of Hyperuricemia (HUA) in elder population of Changchun city,and to detect the correlation between cardiovascular risk factors and the HUA.Methods 900 residents older than 55 years were selected randomly for this questionnaire survey.Physical and laboratory examinations were performed.Results The HUA prevalence rate elder people in Xixin District of Changchun was 16.0％(144/900),while the rates were 13.7％(50/365),15.2％(47/309) and 20.8％(47/226) (P＜0.05) in the elder group (55-65 years),the aged group (66-75 years),and the advanced aged group (older than 76 years) respectively;there was no statistical significant difference in the prevalences between male and female (x2=0.023 5,P＞0.05).The HUA prevalence rate was significantly different between people who had different level of blood pressure,cholesterol,hypersensitive C-reactive protein (hs-CRP),body mass index (BMI),waisthip ratio (WHR).The level of uric acid (UA),total cholesterol (TC) and hs-CRP was significantly different in people with high uric acid when compared with those of normal patients (P＜0.05).There was positive correlation between UA level and TC,triglyceride (TG) level (r=0.364,P＜0.05;r=0.479,P＜0.05).Conclusion The HUA prevalence rate increases significantly as people getting older.There is positive correlation between the increase of uric acid level and the major cardiovascular risk factor.People with hypertension,hyperlipidemia,overweight and obese have high risk for HUA,so change life style and dietary habits may prevent or reduce the occurrence of HUA.

Objective To observe the clinical efficacy of modified Sanzi Yangqin Decoction for the treatment of obstructive sleep apnea-hypopnea syndrome (OSAHS). Methods Totally 80 cases of OSAHS patients were randomly divided into treatment group and control group, with 40 cases in each group. Both groups received intervention of diet and life. The control group was given vitamin C, 100 mg each time, 3 times a day orally. Treatment group was given modified Sanzi Yangqin Decoction, 1 dosage per day, twice a day, orally, for 14 d. The scores of TCM symptoms, sleep apnea (AHI), lowest oxygen saturation (LSaO2) and longest apnea were observed before and after treatment. Results The overall effective rate of TCM syndrome was 90% (36/40) in the treatment group and 65% (26/40) in the control group, with statistical significance (P<0.05). Compared with before treatment, there was statistical significance in the scores of TCM symptoms, AHI, LSaO2, and longest apnea (P<0.05,P<0.01). After the treatment, compared with the control group, there was statistical significance in the scores of TCM symptoms, AHI, and LSaO2 in the treatment group (P<0.05,P<0.01). Conclusion Modified Sanzi Yangqin Decoction can effectively treat OSAHS and improve the life quality of patients.

A rapid, sensitive and convenient LC-MS/MS method was developed for the determination of γ-aminobutyric acid (GABA) in human plasma. d2-γ-Aminobutyric acid (d2-GABA) was synthesized as internal standard (IS). After extraction from human plasma by protein precipitation with acetonitrile, all analytes were separated on a Luna HILIC column (100 mm x 3.0 mm, 3 μm) using an isocratic mobile phase of water: acetonitrile: formic acid (20 : 80 : 0.12) with a flow rate of 0.5 mL x min(-1). Acquisition of mass spectrometric data was performed in multiple reaction monitoring mode (MRM) in positive electrospray ionization using the transitions of m/z 104 --> 69 for GABA and m/z 106 --> 71 for d2-GABA. The method was linear in the concentration range of 5.00 to 1 000 ng x mL(-1). The intra- and inter-day precisions were within 9.9%, and accuracy ranged from 99.1% to 104%, within the acceptable limit across all concentrations. The method was successfully applied to a pharmacokinetic study of GABA tablets in healthy Chinese volunteers.

Objective To investigate the characteristics of the patients with post-stroke depression (PSD).Methods208 cases of patients with stroke were analyzed retrospectively. Results and ConclusionThe PSD occurred more frequently within the first month but less on the first week (P<0.001). The incidence of PSD was higher in female than in male (P<0.001).Frontal, temporal, parietal cortices lesion significantly increased the incidence of the PSD compared with in other area(P<0.01). The more severely the neural function was damaged, the higher the incidence of the PSD was. The muscle strength of patients with PSD recovered poorer than those without PSD. There was significant difference in the incidence of the PSD between the middle age group and aged group (P<0.01).