Objective The present study is to investigate the influence of vaginal discharge on dry chemistry determination of leukocyte esterase in female urine.Methods Collected 20 and 30 normal vaginal discharge samples.Which humoral routine test degree were Ⅰ and Ⅱ correspondingly,and then analyze leukocyte esterase and squamous epithelial cells in these normal vaginal discharge samples.Collected normal vaginal discharge samples again and isolate the vaginal squamous epithelial cells from those normal vaginal discharge samples.Made two concentration of squamous epithelial cell suspension liquid (20/μl and 60/μl)and added these liquid.To normal female urine and analyze the drying chemical leukocyte esterase in those urine. Cleaned the vulva of those patients with normal leukocyte counts in urine sediment determination,whose dry chemical deter-mination of leukocyte esterase was strong positive,perform the routine urine test with her middle urine.Results The results of the determination of leukocyte esterase in normal vaginal discharge samples were 66.7% positive.And there were ~squamous epithelial cells in all samples (microscopy).No significant difference was observed in the examination of dry chem-istry leukocyte esterase among the normal female urine group,low (20/μl)and high (60/μl)concentrations of squamous epi-thelial cell urine group (P >0.05).The counts of squamous epithelial cells and the rate of positive and intensity of dry chem-ical leukocyte esterase in the middle of second urine were significantly lower than before,and the differences are statistically significant (P <0.05).But there was no significant difference for leukocytes counts.Conclusion Urine squamous epithelial cells had no influence on the detection of leukocyte esterase by dry chemistry.However,the leukocyte esterase in the vaginal discharge greatly influenced the examination of the leukocyte esterase in urine.

Objective To investigate the distribution of subtypes of T. pallidum (TP) in Hengyang and Jiangmen regions. Methods Eighty-five specime ns taken from patients with suspected chancre collected in Hengyang and Jiangmen from February 2002 to January 2004 were screened by a PCR targeted TP polA gene , and then the arp gene and tpr gene were amplified from TP positive specimens. The PCR products of tpr gene were digested by restriction endonuclease MSe I. Th e sizes of the arp gene and the restriction fragment length polymorphism (RFLP) of the tpr gene were analyzed for subtyping. Results Of 69 TP-positive specime ns, 57 could be subtyped, and 10 subtypes were found. Among them, 26 (45.6%) wer e subtype 14d, and other subtypes included 10d(1), 12a(3), 12g(2), 13d(6), 14a(5 ), 14b(2), 14f(6), 15d(5) and 16d(1). No significant difference of the distribut ion of TP subtypes between Hengyang and Jiangmen was found. Conclusion Multipl e T.pallidum subtypes have been prevalent in Hengyang and Jiangmen, although the predominant subtype is 14d, there is no significant geographic heterogeneity be tween these two regions.

Objective To evaluate the efficacy of stomach intestine power regulator, intestinal microecology preparation and tricyclic antidepressant treatment in irritable bowel syndrome (IBS), and to investigate its pathological mechanism. Methods From November 2006 to November 2010, 103 patients with diarrhea-dominant IBS (D-IBS), who fulfilled the Rome Ⅱ criteria and were excluded from organic disease by entewscope were divided into pinaverium bromide group (26 cases), pinaverium bromide + bifid triple viable group (28 cases), pinaverium bromide + doxepin group (25 cases) and pinaverium bromide +bifid triple viable + doxepin group(24 cases ) by random digits table. The symptom grade, intestinal flora and SCL-90 was tested before treatment and 4 weeks after treatment. Results The total effective rate of pinaverium bromide + bifid triple viable + doxepin group was 83.33%(20/24), significant higher than that in pinaverium bromide group [65.38%(17/26)], pinaverium bromide + bifid triple viable group [71.43%(20/28)], pinaverium bromide + doxepin group [68.00% ( 17/25 )] (P < 0.05 ). Five kinds of intestinal flora and psychiatric symptoms were improved in the four groups, and those in pinaverium bromide + bifid triple viable + doxepin group improved significantly. Conclusions To interfere the correlation factor of IBS can have better efficacy. There is a close relation between brain and gut in patients with IBS, which may be involved in the pathogenesis of IBS.

Objective To investigate the effect of 5-hydroxytryptamine serotonin receptor-1A (5-HT1 A) agonists on micturition dysfunction in rats with chronic spinal cord injury (SCI).Methods Female SD rats weighing 175 -200 g were used.Seven of the rats were modified for a spinal cord injury model (transsection at T10).Eight weeks later,control rats and SCI rats were tested.Rats were anesthetized with urethane ( 1.3 g/kg ).A polyethylene (PE) -50 catheter was placed in the left jugular vein for intravenous drug administration.A PE-90 catheter was inserted through the bladder dome,and the other end of the bladder catheter was connected to a syringe pump for continuous infusion of saline and to a pressure transducer for intravesical pressure monitor.Dose-response curves for 8-OH-DPAT were followed by the WAY-100635 test.The capacity,residual volume,micturition volume,and EUS-EMG were measured.Results With an increasing dose of 8-OH-DPAT,the capacity of the bladder decreased from 33.2 ± 8.3 ml to 22.8 ± 2.4 ml.The micturition volume was increased from 0.14 ± 0.08 ml to 0.38 ± 0.09 ml.The residual volume decreased from 3.68 ± 1.36 ml to 1.84 ± 0.21 ml,and peak intravesical pressure changed from 27.1 ± 3.6 mm Hg to 22.8 ± 2.4 mm Hg.Control rats showed little significant change in the cystometric variable.Effects of 8-OH-DPAT were reversed by WAY-100635.That 8-OH-DPAT induced phasic relaxation occured in spinal cord-injured rats but the control group showed no significant change.ConclusionsThe 5-HTIA/7 receptor agonist 8-OH-DPAT may induce periodic EUS relaxation during voiding in urethane-anesthetized chronic spinal cordinjured rats.And this could result in an increase in micturition volume,a decrease in bladder capacity,and thus an increase in voiding efficiency.

BACKGROUND: Some studies suggest that pre-injection of tumor necrosis factor-α (TNF-α)can protect focal cerebral ischemia in mice. Cerebral ischemia tolerance is related to the increase of TNF-α level; on the other hand, TNF-α is an injurious cytokine associated with stroke. Circulating antibody against anti-TNF-α can protect reperfused injury.OBJECTIVE: To study the effects of TNF-α pretreatment and post-treatment on cerebral ischemia-reperfusion injury and explore possible mechanism.DESIGN: Randomized controlled study.SETTING: Neurological Department, the Second Hospital Affiliated to Harbin Medical University.MATERIALS: The experiment was conducted at the Animal Experiment Center of Harbin Medical University from January to April 2002. Totally 120 healthy adult male Wistar rats were randomly divided into the following 8 groups: TNF-α 0.05 μg, 0.5 μg and 1.0 μg pretreatment groups and PBS group, TNF-α 0.05 μg, 0.5 μg and 1.0 μg post-treatment groups and PBS group with 15 in each group.METHODS: The focal brain ischemia model of middle cerebral artery occlusion (MCAO) was made using inserting thread method. TNF-α of different doses (0.05 μg, 0.5 μg or 1.0 μg) or PBS was injected intracisternally and 22-hour reperfusion, 8 rats from each group were killed. Then the perhour reperfusion, 7 rats from each group were killed. Then pathological changes were observed, glial fibrillary acidic protein (GFAP) and intercellular adhesion molecule-1 (ICAM-1) expression were inspected by immunohistochemical method. Histopathological and immunohistochemical evaluation was made with the computer-assisted image analyzing system,and the number of GFAP positive cells and ICAM-1 positive vessels in each hemisphere was counted.riliary acidic protein and ICAM-1.infarct volume: TNF-α 0.5 μg and TNF-α 1.0 μg pretreatment groups showed reduced volume of lesion; infarct volume reduced by 70.9% in TNF-α 0.5 μg pretreatment rats and 66.5% in TNF-α 1.0 μg pretreatment rats. TNF-α 0.5 μg and TNF-α 1.0 μg post-treatment groups showed increased volume of lesion; infarct volume increased by 22.3% in TNF-α 0.5 μg post-treatment rats and 46.7% in TNF-α 1.0 μg post-treatment rats.TNF-α 0.05 μg and 1.0 μg pretreatment groups did not differ significantly (P ＞ 0.05), but there was an obvious difference between TNF-α 0.5 μg and pared with PBS pretreatment group, TNF-α 0.5 μg and 1.0 μg pretreatment groups showed lessened tissue damage and edema. Compared with PBS post-treatment group, TNF-α 0.5 μg and TNF-α 1.0 μg post-treatment fibriliary acidic protein and ICAM-1: TNF-α 0.5 μg and TNF-α 1.0 μg pretreatment groups showed reduced volume of glial fibriliary acidic protein and ICAM-1 (P ＜ 0.05); but TNF-α 0.5 μg and TNF-α 1.0 μg posttreatment groups showed increased volume of glial fibriliary acidic protein and ICAM-1 (P ＜ 0.05). TNF-α 0.05 μg and 1.0 μg pretreatment groups did not differ significantly (P ＞ 0.05); but there was an obvious difference between TNF-α 0.5 μg and 1.0 μg post-treatment groups (P ＜ 0.05).cerebral ischemia reperfusion injury. This effect is not related to the repair given after cerebral ischemia reperfusion, ischemia exacerbates, which is α are determined by whether TNF-αis given before or after cerebral ischemia in a dose-dependent manner.

Objective:To investigate the influence of lumbar disc degeneration on the efficacy of lumbar fixed-point rotation manipulation in sitting position in treating lumbar intervertebral disc herniation (LIDH). Methods:We simulated lumbar fixed-point rotation manipulation of sitting position using three finite element models including a normal model, a mild degeneration model and a moderate degeneration model of L3-5, in which the herniated disc was assumed at the left rear of L4 disc and the rotation manipulation was carried out on the right side. The displacement and stress at the left rear of L4 disc of the three models were analyzed. Results:When lumbar fixed-point rotation manipulation in sitting position was carried out, a displacement and stress were generated at the left rear of L4 intervertebral disc of the three models directing forward. The displacement and stress in degeneration models were less than those in the normal model, and the smallest values were found in the moderate degeneration model. From normal model to mild and then to the moderate degeneration model, the displacement decreased by 36% and 59%, and the stress decreased by 22.3% and 45.2%, respectively. Conclusion:The lumbar disc degeneration affects adversely the effectiveness of lumbar fixed-point rotation manipulation in sitting position in the treatment of LIDH. The severer the lumbar degeneration, the greater the influence.

Objective To evaluate the cellular toxic and release of pharmaceutical dressing.Method Following the State standard GB/T14233.2-2005.the L929 cellular merphology was observed by inveded microscopy after 72h and proliferation of the cells was examined using mitochondrial function(MTT)assay.Relative growth rate (RGR)was calculated and cytotoxicity grade was evaluated by absorbency(OD)data.With PBS7.4 as dissolution media,and(32±0.5)℃as dissolution temperature,the release rate was determined with UV method with the determination wavelength of 288 nm and the dissolved liquid in 1/6,1/2,1,3,16,24,36 and 48 h.Results The average cell RGR of the pharmaceutical dressing is 91.25%and reaches 1 cytotoxicity grade.L929 cellular morphology is normal.Pharmaceutical dressing release accord to Higuchi equation,and the simulated equation is M_t/M_∞=0.3271t~(0.239).Conclusions Biologic compatibility of the pharmaceutical dressing is good,and the release of levofloxacin from the pharmaceutical dressing is sustained in vitro.

Objective To investigate the effect of acupuncture combined with minimally invasive hematoma aspiration on a rabbit model of brain injury in the acute stage of intracerebral hemorrhage (ICH). Methods A total of 65 healthy New Zealand white rabbits were randomly divided into a sham operation (SO) group, an ICH group,a hematoma aspiration (HA) group, and an acupuncture and hematoma aspiration (AHA) group. Models of ICH were established in the latter three groups. The HA group was treated with minimally invasive hematoma aspiration and the AHA group was treated with both acupuncture and minimally invasive hematoma aspiration. At 6 hours and 1, 3 and 7 days after the ICH models were established, brain water content (BWC) was measured, and the expression of matrix metalloproteinase-9 ( MMP-9 ) in the rabbits' brains was detected by immunohistochemistry. MMP-9activity was detected by gelatin zymography. Results The BWCs of the ICH group, HA group and AHA group rabbits were significantly higher than those of the SO group. The BWCs of the HA group and AHA group animals descended significantly more than those in the ICH group as time went on, especially on the 3rd day in the AHA group. Immunohistochemistry and gelatin zymography showed that the expression and activity of MMP-9 in these test groups decreased with time, especially on the 3rd day in the AHA group. Conclusions Acupuncture combined with hematoma aspiration can reduce injury in the acute stage of ICH, and inhibition of the expression of MMP-9 may be the mechanism.

Objective To evaluate the effects of morphine preconditioning on the expression of microRNAs (miRNAs) during hypoxia-reoxygenation (H/R) in isolated cardiomyocytes in rats with heart failure.Methods Healthy adult male Sprague Dawley rats,w eighing 200-220 g,were used in this study.Adriamycin 2.0 mg/kg was injected once a week for 6 weeks via the tail vein to induce heart failure.The cardiomyocytes were isolated from the failing hearts of rats and seeded in 24-well plates or in 60 mm diameter dishes.The cells were then randomly divided into 3 groups (n =16 each) using a random number table:control group (group C); group H/R;morphine preconditioning group (group MP).The cells were cultured in normal culture atmosphere in group C.After being exposed to hypoxic air (5％ CO2-95％ N2) for 90 min,the cells were returned to the high-glucose DMEM supplemented with 10％ newborn bovine serum and were then cultured for 120 min in H/R and MP groups.In group M,the cells were cultured in morphine culture medium (final concentration of morphine 0.3 μmol/L) for 10 min and then were returned to the culture medium without morphine and cultured for 30 min immediately before hypoxia.At 120 min of reoxygenation,the cells of 8 wells in each group were chosen to detect the cell viability and lactate dehydrogenase (LDH) activity (by Typan blue staining).All the RNAs were extracted from the cardiomyocytes of the left 8 wells in each group and subjected to miRNA microarray to screen differentially expressed miRNAs.Results The cell viability was significantly lower,the activity of LDH was higher,the expression of miR-6216 and let7e-5p was higher,and the expression of miR-133b-5p was lower in H/R and MP groups than in group C (P ＜ 0.05).Compared with H/R group,the cell viability was significantly increased,the activity of LDH was decreased,the expression of miR-133b-5p was up-regulated,and the expression of miR-6216 and let-7e-5p was down-regulated in MP group (P ＜ 0.05).Conclusion Morphine preconditioning reduces H/R injury to isolated cardiomyocytes in rats with heart failure through regulating the expression of miRNAs such as miR133b-5p,miR-6216 and let-7e-5p.

Objective To evaluate the roles of 1-phosphatidylinositol 3-kinase (PI3K) and extracellular signal-regulated kinase (ERK) signaling pathways in reduction of ischemia-reperfusion (I/R) injury to the isolated hearts by morphine preconditioning in the rats with chronic heart failure.Methods Adult male Sprague-Dawley rats,weighing 200-230 g,in which doxorubicin 2.0 mg/kg was injected via the tail vein once a week for 6 weeks to induce chronic heart failure,were studied.At the end of 8th week,42 rats with chronic heart failure were randomly divided into 7 groups (n =6 each) using a random number table:sham operation group (group S),I/R group,morphine preconditioning group (group MP),PD98059 (ERK inhibitor) + morphine preconditioning group (group PD + MP),wortmannin (PI3K inhibitor) + morphine preconditioning group (group WT + MP),PD98059 group (group PD) and wortmannin group (group WT).The hearts were quickly excised and passively perfused in a Langendorff apparatus and subjected to 30 min of occlusion of the left coronary artery followed by 2 h of reperfusion to establish the model of I/R injury.In group S,the hearts were only sutured,but not ligated and were continuously perfused with K-H solution for 195 min.In group I/R,the hearts were perfused with K-H solution for 45 min before ischemia.In group MP,the hearts were perfused with K-H solution for 15 min,with K-H solution containing morphine 1 μmol/L for 5 min and then with K-H solution for 5 min (3 cycles in total) before ischemia.In PD + MP and WT + MP groups,the hearts were perfused with K-H solution containing PD98059 (10 μmol/L) and wortmannin (100 nmol/L),respectively,starting from 10 min before morphine preconditioning until 5 min of ischemia.In PD and WT groups,the hearts were perfused with K-H solution containing PD98059 (10 μmol/L) and wortmannin (100 nmol/L),respectively,starting from 40 min before ischemia until 5 rin of ischemia.At 15 min of equilibration (baseline) and 5 and 10 min of reperfusion,the coronary flow was collected to detect the activity of lactate dehydrogenase (LDH).Infarct size (IS) and area at risk (AAR) were measured at the end of reperfusion and IS/AAR ratio was calculated.Results Compared with group S,LDH activity was significantly increased at 5 and 10 min of reperfusion,IS and IS/AAR ratio were also increased (P ＜ 0.05),and no significant change was found in AAR in group I/R (P ＞ 0.05).Compared with group I/R,LDH activity was significantly decreased at 5 min of reperfusion,IS and IS/AAR ratio were also decreased (P ＜ 0.05),and no significant change was found in AAR in group MP,and no significant change was found in LDH activity,IS,AAR and IS/AAR ratio in WT and PD groups (P ＞ 0.05).Compared with group MP,LDH activity was significantly increased at 5 and 10 min of reperfusion (P ＜ 0.05),and IS and IS/AAR ratio were decreased in group PD + MP,and no significant change was found in LDH activity,IS,AAR and IS/AAR ratio in group WT + MP (P ＞ 0.05).Conclusion Activation of ERK signaling pathway is involved in reduction of I/R injury to isolated hearts by morphine preconditioning in rats with chronic heart failure,however,PI3K signaling pathway has no such effect.