Objective To validate and compare the two spinal microdialysis techniques: a linear tissue probe (LM-3) in the spinal dorsal horn and a loop probe in the cerebral spinal fluid (CSF) in determining peripheral nociceptive stimulation-evoked glutamate (Glu) release in the spinal cord of freely moving rats. Methods Twenty-eight adult male Wistar rats weighing 300-350 g were randomly divided into two groups: in group A a LM-3 probe was implanted into the spinal dorsal horn and in group B a loop probe was placed in the CSF. Twenty-four hours after the implantation of the probe, microdialysis was initiated with perfusion of modified Ringer' s solution at a low flow rate of 5 ?l?min-1 . Following an 1 h equilibration phase the baseline Glu concentrations were measured every 10 min for 1 h. Thereafter 50 ?lof 5% formalin was injected into one hindpaw of the rats and samples were collected every 10 min for 90 min. Furthermore 8 rats in group A were further divided into 2 subgroups to investigate the effects of the flow rate of microdialysis and composition of perfusate on the baseline Glu release.Results The baseline levels of Glu were (0.82?0.09) ?mol?L-1 with LM-3 probe and (5.96?0.22) ?mol?L-1 with the loop probe. In group A (LM-3 probe) when the flow rate of the modified Ringer's solution was decreased from 5 to 2 ?l?min-1 the extracellular Glu concentrations were increased to 223%?7% of the baseline (n = 4) , whereas perfusion with artificial CSF reduced Glu concentrations to 62% ?10% of the baseline (n = 4) . Injection of formalin into the hindpaw induced a short-lasting but significant increase in Glu concentration with a similar profile and time course using either of the two microdialysis approaches. Conclusion Microdialysis in the dorsal horn or in the CSF are both effective techniques to assess Glu release in the spinal cord of rats. Peripheral nociceptive input induces a short-lasting increase in Glu release with a similar profile and time course using either of the two microdialysis approaches. The microdialysis of the dorsal horn provides a useful tool to precisely locatewhere the release of the neurotransmitter occurs, whereas the loop probe in CSF is more reproducible for simultaneous investigation of drug effects.

BACKGROUND: Xingnao kaiqiao acupuncture is a series of systematic acupuncture methods with yin channels in domination, assisted by specified standards of quantity manipulation, which was founded according to the pathogenesis of stroke, and it is effective in treating stroke.OBJECTIVE: To observe the effects of xingnao kaiqiao acupuncture on blood lipids and blood viscosity in stroke patients of different syndrome types.DESIGN: A comparative observation.SETTINGS: Institute of Acupuncture and Moxibustion, and Special Care Ward of Acupuncture, the First Affiliated Hospital of Tianjin College of Traditional Chinese Medicine.PARTICIPANTS: Totally 750 stroke inpatients (466 males and 284 females) were selected from the Special Care Ward of Acupuncture, the First Affiliated Hospital of Tianjin College of Traditional Chinese Medicine between January and December in 2001, the average age was (64±12) years,the disease course ranged from 2 hours to 3 years. All the patients were informed and agree with the therapeutic program.METHODS: ① The traditional Chinese medicine (TCM) syndrome differentiations of stroke were in reference to the fifth edition of Traditional Chinese Medicine and Internal Medicine. There were 688 cases of stroke of meridian and collateral type and 47 cases of stroke attacking zang-fu organs type. ② Huatuo filiform needles produced by Suzhou [1.0-1.5 cun in length, 0.32-0.38 mm in diameter (No. 30-28)] were used in the xingnao kaiqiao acupuncture. Firstly, the acupoints of bilateral Neiguan were acupunctured perpendicularly for about 0.5-1.0 cun, the reducing manipulation by twirling as well as lifting and thrusting the needle was performed for 1 minute; Secondly, the needle was inserted at Renzhong towards nasal septum for about 0.3-0.5 cun, the manipulation of heavy bird-peck needling was used till the eyeballs were wet or tears fell down; Then Sanyinjiao was acupunctured, the needle was inserted along the medial border of tibia at an angle of 45° with skin for about 1.0-1.5 cun, the reinforcing manipulation by lifting and thrusting the needle was performed till the ipsilateral lower limb had three tics; The acupoint of Weizhong was selected by lying and lifting the leg, the needle was inserted perpendicularly for 0.5-1.0 cun, and the reducing manipulation by lifting and thrusting was applied till the ipsilateral lower limb had three tics; The acupoint of Jiquan was acupunctured at 1 cun below the source point along the channel in avoidance of axillary hairs, the needle was inserted perpendicularly for 1.0-1.5 cun, and the reducing manipulation by lifting and thrusting was applied till the ipsilateral upper limb had three tics; The elbow was flexed to 120° for acupuncture at Chize, the needle was inserted perpendicularly for 1.0 cun, the reducing manipulation by lifting and thrusting was applied till the ipsilateral upper limb had three tics. Twice a day for 7 days as a course. ③ After four courses averagely, the whole blood viscosities were detected with the hemorrheologic detector, and the normal ranges for lowshear, middle-shear and high-shear of whole blood viscosities were 6.50-9.25, 4.35-5.45 and 3.65-4.40 mpa ·s respectively. The levels of total cholesterol and triglyceride were detected with blood lipid detector, and the normal ranges were 3.38-6.5 mmol/L and 0.56-0.147 mmol/L.MAIN OUTCOME MEASURES: The effects of xingnao kaiqiao acupuncture on the blood lipids and blood viscosity in stroke patients of different syndrome types were observed.RESULTS: Totally 690 and 721 cases finished the detections of whole blood viscosities and blood lipids respectively. There were obviously fewer cases with normal whole blood viscosities in the patients with stroke of meridian and collateral than in those with stroke attacking zang-fu organs (10.3%, 15.4%, P ＜ 0.01), but markedly more cases with higher total cholesterol and triglyceride than in those with stroke attacking zang-fu organs (80.5%, 91.5%; 64.1%, 71.3%; P ＜ 0.01).CONCLUSION: The levels of blood lipids in the patients with stroke of meridian and collateral are more abnormal than in those with stroke attacking zang-fu organs, but their blood viscosities are better than in those with stroke attacking zang-fu organs.

Objectives To study the effect of highway transportation on the quality of red blood cells. Methods Highway transportation environment was simulated by random vibration test. Red blood cell suspensions and whole blood were vibrated for 120 minutes based upon GB/T 4857.23-2003 with the RMSVA of 0.38 and 0.52grms, as well as GJB 2711-1996 with the RMSVA of 1.04grms and weight loading of 60kg. Five ml of specimen were taken before the testing (as Control Group), and after the testing at 30mins' intervals (as Testing Group). K+, FHb and red blood cell osmotic fragility were tested. Results The red blood cell osmotic fragility of whole blood increased in 0.38grms, 90min and 0.52grms, 60min specimens (P

Objective To study the hydrogen sulfide productivity by mononuclear cells in patients with atrial fibrillation (AF). Methods Mononuclear cells were extracted from peripherial blood of 60 AF patients and 30 healthy controls. Modified methylene blue method was used to evaluate the hydrogen sulfide productivity of mononuclear cells. Results Hydrogen sulfide productivity of mononuclear cells from AF patients was 92.5%higher than that from control [10.63 (8.21~14.37) μmol/(min.g protein) vs 5.52 (4.42~8.50) μmol/(min.g protein)]. Hydrogen sulfide productivity of mononuclear cells was correlated with the left atrial transverse diameter (r = 0.276,P < 0.01), while no significant correlation was found between hydrogen sulfide and left ventricular end-diastolic diameter , left atrial anterior posterior diameter , C-reaction protein etc. Conclusion Hydrogen sulfide productivity of mononuclear cells from AF patients is promoted.

Objective To investigate the results of tumor specific growth factor(TSGF) levels in in healthy people and cancer patients in Guangzhou areas .Methods The serum TSGF levels of 6 615 health check-up receivers and 287 patients with cancer in the hospital from July 2012 to June 2013 were detected .Results In health check-up receivers ,the level and positive rate of TSGF were higher in >40-50 age women .The level and positive rate of TSGF in men were increased with age .The levels of TSGF in pa-tients with lung cancer ,liver cancer and digestive system tumor were higher than that in health check-up receivers(P<0 .01) ,but there has no statistically difference between genital system cancer patients and healthy check-up receivers(P>0 .05) .Conclusion Performing TSGF test in health examination is great significance to the early diagnosis of malignant tumors .The reference value range of TSGF should be set according to different gender and age .

OBJECTIVE:To investigate the correlation of gene polymorphism in peripheral artery disease(PAD)patients with antiplatelet efficacy of clopidogrel. METHODS:Reviewing related domestic and foreign literatures in recent years,the correlation of gene polymorphism in PAD patients with antiplatelet efficacy of clopidogrel was summarized and analyzed. RESULTS&CON-CLUSIONS:At present,a variety of genes associated with clopidogrel antiplatelet efficacy and major adverse cardiovascular events (MACE)have been identified,including cytochrome P450(CYP)2C19,adenosine three phosphate binding cassette B subfamily 1 (ABCB1),paraoxonase 1 (PON1) and adenosine diphosphate P2Y12 receptor (P2Y12),etc. CYP2C19*2,*3 allele may reduce the antiplatelet effect of clopidogrel. Their correlation has been confirmed by a number of studies,and study results are broadly con-sistent. Mutations in the ABCB1 C3435T and PON1 Q192R sites may lead to a lower response to clopidogrel and increase the risk of MACE;but there is a lack of large-scale prospective clinical studies,and the present results are inconsistent. P2Y12 gene poly-morphism in PAD patients has not been found to be significantly associated with clopidogrel efficacy.

Objective To investigate the role of self-made auxiliary device in pelvic tumor radiotherapy with phantom immobilization using Varian cone-beam CT (CBCT).Methods A total of 50 patients with pelvic tumor were enrolled and randomly divided into study group and control group according to the order of enrollment.The patients in the study group were immobilized with thermoplastic phantom and self-made auxiliary device,and those in the control group were immobilized with thermoplastic phantom.CBCT scan and online matching were regularly performed before radiotherapy to obtain the setup errors of the left-right (x),cranial-caudal (y),and anterior-posterior (z) directions.The independent-samples t-test was used for comparison between groups.Results The set-up errors in the x-,y-,and z-directions in the study group were 1.56± 1.00 mm,1.60± 1.29 mm,and 1.36± 1.00 mm,respectively,and those in the control group were 1.76±1.33 mm,2.76±1.69 mm,and 1.92±0.91 mm,respectively (P=0.551,0.009,and 0.043).Conclusions Self-made auxiliary device helps to eliminate the errors in the cranial-caudal direction and solve the problem of involuntary activities of the lower limbs.

Objectives To analyze the expression of fibroblast growth factor-19(FGF-19) in hepatocellular carcinoma ( HCC) and adjacent tissues , and to investigate its clinical significance .Methods A total of 209 HCC patients who had undergone radical resection operations at Hospital 401 between January 2003 and December 2009 were chosen as samples . Immunohistochemistry method was employed to examine the expression level of FGF-19 in HCC and adjacent tissues .The relationship between FGF-19 protein expressions and clinicopathological features was analyzed by the chi -square test or Fisher exact probability .A survival curve was drawn using the Kaplan-Meier method and the Cox model was used to analyze factors that influenced survival .Results The rate of high expression of FGF-19 was 66.1% (138/209) in HCC, which was significantly higher than 46.9%(98/209) in adjacent tissues (P<0.05).The high expression of FGF-19 was related to the tumor capsule and tumor boundary (P<0.05).The overall survival in high expression of FGF-19 group was signifi-cantly lower than that in low expression group (P<0.05).Conclusion FGF-19 plays an important role in the carcinogen-esis and development of HCC , and a high expression of FGF-19 might be closely related to survival time of postoperative patients.FGF-19 might be a potential prognosis prediction factor for HCC .

Objective To construct a recombinant plasmid DNA containing herpes simplex virus type 1(HSV-1) glycoprotein D (gD) gene.Methods The HSV-1 gD gene was obtained by polymerase chain reaction (PCR) and inserted into TA cloning vector pGEM-T, then cloned into the eukaryotic expression vector pcDNA3.1 to generate pLy-D. The recombinant plasmid pLy-D, which was confirmed by partial sequencing and restriction endonuclease analysis, was transfected into Cos-7 cells and used to inoculate ICR mice via muscular injection. Immunohistochemistry and enzyme-linked immunoabsorbent assay (ELISA) were employed to test the gD expression in transfected cells and the specific anti-HSV-1 antibody in the serum of immunized mice, respectively.Results The gD eukaryotic expression plasmid pLy-D was constructed. Using the immunohistochemistry technique, the gD expression in pLy-D-transfected cells was detected. The ELISA demonstrated that specific anti-HSV-1 antibody could be induced in immunized mice after three times injection.Conclusions We constructed HSV-1 gD eukaryotic expression plasmid pLy-D which could express gD protein in transfected cells and could induce humoral immune response in mice. This observation will be helpful in designing HSV prophylactic vaccine.

Objective To evaluate the role of 5-HT5A receptors (5-HT5A R) in activation of astroglia in the spinal dorsal horn in a rat model of neuropathic pain induced by vincristine. Methods Forty adult male SD rats weighing 180-200 g were randomly divided into 4 groups ( n = 10 each): control group (group C);neuropathic pain group (group P);Ad-X-HK group (group B) and Ad-5-HT5A-siRNA group (group S). Neuropathic pain was induced by repeated intraperitoneal (IP) injection of vincristine 0.1 mg/kg according to the method described by Weng et al in group P, B and S. On the 2nd day after the last IP injection, the animals received artificial cerebrospinal fluid, Ad-X-HK and Ad-5-HT5A-siRNA 25 μl administered intrathecally (IT) in group P, B and S respectively. Paw withdrawal threshold to mechanical stimulus was measured before and on the 7th day after IT administration. The animals were then sacrificed. The lumbar segment ( L4.5 ) of the spinal cord was removed for determination of 5-HT5A R and GFAP expression. Results Body weight and paw withdrawal threshold were significantly decreased after repeated IP vincristine administration in group P compared with group C. IT Ad-5-HT5A-siRNA reduced pain threshold further in group S compared with group P. Repeated IP vincristine significantly increased the expression of 5-HT5A R and GFAP in spinal dorsal horn, and IT Ad-5-HT5A-siRNA significantly decreased the expression of 5-HT5A R while increased the expression of GFAP in spinal dorsal horn in group S compared with group P. Conclusion 5-HT5AR is involved in the inhibition of astrocyte activation, resulting in reduction of vincristineinduced neuropathic pain.