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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 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 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).

Objective To investigate the difference of endothelial cell (EC) in ultrastructure,markers and permeability between blood-brain barrier (BBB) and blood-nerve barrier (BNB).MethodsThe rat brain cortex and sciatic nerve were removed.Ultrastructure of endothelial cell was observed with electron microscopy.Employing immunohistochemical (IHC)staining,the relative distribution was determined for the OX-26,endothelial barrier antigen (EBA)and extravasated fibrinogen around microvessels.ResultsMicrovessels both in BBB and in BNB did share the same tight junctions.However,BNB had a significantly larger number of vesicles than that of BBB.Microvessels in BNB did not express the OX-26 and EBA,but staining weakly positive for fibrinogen,while microvessels in BBB showed strong positive for OX-26 and EBA,perivascular staining of fibrinogen had not been seen.ConclusionTight junctions were present in both BBB and BNB endothelial cell,but there was a significant difference between them in the number of the pinocytotic vesicles,markers expression and permeability.

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.

Objective To explore clinical curative effects of the moist burn cream combined with ozone in treatment of fat liquefaction of abdominal incision after appendectomy.Methods 112 patients who suffered with fat liquefaction of abdominal incision after appendectomy in hospital from January 2012 to January 2015 were chosen.According to the random number table method, they were randomly divided into the observation group and control group, 56 cases in each group.Before treatment, two groups of patients were liquefied suture removal and thoroughly removed subcutaneous tissue necrosis.Then, the control group accepted the pure moist burn cream treatment, while the observation group were given the moist burn cream combined with ozone treatment.The growth condition of granulation tissue after treatment, frequency of dressing change, wound cicatrisation, and length of hospital stay were compared between two groups, and the condition of wound healing was evaluated.Results The grade A healing rate of the observation group was 89.28%(50/56), while that of control group was 71.43%(40/56)(χ2 =5.65,P<0.01).The time of the granulation tissue beginning to grow and the time when the granulation tissue flushed leather face of observation group were significantly shorter than those of control group ( P<0.05 ) .In addition, times of changing a medical prescription of observation group was less than that of control group (P<0.05), and the incision healing time, length of stay in hospital were also shorter than those of control group (P<0.05).Conclusion It has better clinical curative effect to apply the moist burn cream combined with ozone in the treatment of fat liquefaction of incision after appendectomy.Moreover, it has some advantages, such as the quick growth of granulation, the well-healed wound, and the short hospitalization time.Therefore, it is worthy of clinical popularization and application.

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.