Objective: To explore the effects of continuous trickle of low temperature liquids through the lateral ventricle on neural cell apoptosis after rabbit local cerebral ischemia/reperfusion. Methods:The middle cerebral artery (MCA) of New Zealand rabbit was clipped by micro aneurysm clip for 2 hours and reperfused for 24 hours. Immediately after clipping the MCA, we trickled the left lateral ventricle continuously with low temperature liquids(22℃) to decrease the brain temperature to mild hypothermia (33℃-35℃)and maintained for 2 hours. After reperfusion for 24 hours , we assessed TUNEL method to determine the apoptotic cell rate in the sham-operated group, the control group and the mild hypothermia group respectively. Results:The apoptotic cell rate of the cortex tissues accommodated by MCA in the mild hypothermia group was obviously lower than that in the control group(P0.05). Conclusion:Trickling ventricle with low temperature liquids could decrease the apoptotic cell rate and alleviate cerebral ischemia/reperfusion injury.

BACKGROUND: Neuroprotective role of hypothermia on cerebral ischemic-reperfusional impairment has been long acknowledged. Since general hypothermia is complicated and unfit for observing postoperative consciousness and neurological function, it is of important significance to explore novel methods of focal cerebral hypothermia.OBJECTIVE: To study the neuroprotective effect of lateral ventricle infusion of low-temperature fluid on ischemic neurons of middle cerebral artery (MAC) occlusion models established on New Zealand rabbits.DESIGN: A randomized case-control study based on experimental animal models.SETTING: Neurosurgical department and pathological department of a general military hospital of Chinese PLA.MATERIALS: This study was carried out at Neurosurgical Laboratory of Nanjing General Hospital, Nanjing Military Area Command of Chinese PLA. Altogether 18 healthy New Zealand male rabbits, weighing from 2. 8 to 3.2 kg, were selected 4 - 6 months after birth, and randomly divided into occlusion group, hypothermia group and control group.INTERVENTIONS: Cerebral focal ischemic-reperfusional model was established on the New Zealand rabbits through MCA occlusion for 2 hours followed by reperfusion for 24 hours.MAIN OUTCOME MEASURES: Scores for neurological function, water content in the left and right brain, pathological changes of nerve cells in the left MCA supplying region.pothermia group, significantly higher than that in occlusion group(7.58 ± 0.58 )( P ＜ 0.01 ), but no significant difference could be observed in contrast with brain was(81.64 ± 0.82)% and (79.26 ± 1.30)% in occlusion and hypothermia groups with significant difference between them( P ＜ 0.05), and it was significantly different between the left side [ (81.64 ± 0. 82 )% ] and opyknosis and deep staining could be observed in nerve ganglion cells in occlusion group under optical microscope, but no obvious pathological changes were observed in MCA supplying brain regions in hypothermia group.CONCLUSION: Permanent infusion of low-temperature fluid into the lateral ventricle plays an important neuroprotective role by attenuating cerebral ischemic-reperfusional impairment and improving post-ischemic neurological functions.

Objective To detect Ureaplasma urealyticum (Uu) and Mycoplasma hominis (Mh) in urogenital tract of healthy elementary and high school students,and to determine the biovars and serotypes of Uu isolates.Methods Urine specimens were collected from 957 healthy school children,including 404 elementary school students,260 junior high school students and 293 senior high school students,and submitted for Uu and Mh culture in selective liquid medium.Thc specimens positive for Uu and (or) Mh culture were subsequently subjected to PCR.The biovars and serotypes of Uu were determined.Results Uu was detected in 4.81％ (46/957) of the school children.Among the 46 Uu positive specimens,32 (69.57％) harbored Ureaplasma Parvum (biovar 1 ),14 (30.43％) Ureaplasma urealytieum (biovar 2); 12 (2.97％) were from elementary school students,including 9(75.00％) cases of Ureaplasma Parvum and 3(25.00％) Ureaplasma urealytieum,8(3.08％) were from junior high school students,including 5 cases of Ureaplasma Parvum and 3 Ureaplasma urealytieum,26 (8.87％) were from senior high school students,including 18 (69.23％) cases of Ureaplasma Parvum and 8 (30.77％)Ureaplasma urealytieum.Mh was positive in 9 (0.94％) school students,including 1 elementary school student,1 junior school student and 7 senior high school students.The detection rate of Uu was significantly higher in females than in males in all the 3 groups (elementary school,junior and senior high school) of students.Conclusions Mh and Uu are commensals in urogenital tract of elementary and high school students without symptoms.Ureaplasma Parvum is the predominant biovar in this population,though Ureaplasma urealytieum accounts for a significant proportion,which implies that Uu is commensal in healthy elementary and high school students,and females are more liable to be colonized than males.

Background Acanthamoeba keratitis is a sort of serious infectious eye disease with high causing-blindness rate.Acanthamoeba keratitis often is misdiagnosed as fungal keratitis or viral keratitis in the early stage.Because conventional clinical diagnosis methods show a low specificity and take a long time,timely treatment often is delayed.Conventional PCR does not apply well because the lesion sample is not enough to extract DNA.However,direct PCR can amplify 18S rRNA conserved sequence of acanthamoeba keratitis without the extraction of DNA.Objective This study was to discuss the feasibility for rapid diagnosis of acanthamoeba keratitis using direct PCR to amplify the gene 18S rRNA fragment.Methods Ten acanthamoeba strains were isolated from 10 eyes with acanthamoeba keratitis in Qingdao Eye Hospital.The sensitivity of the direct PCR assay was tested using different numbers of amoebas.The specificity of the assay was tested using DNA extracted from acanthamoeba,candida albicans,pseudomonas aeruginosa,herpes simplex virus-1 (HSV-1) and normal human corneal epithelial cell.Acanthamoeba keratitis models were established using infected method in clean 6-week-old female BALB/c mice.Corneal lesion samples were obtained 1 day,3,5,7,10,15 days after modeled.The effectivity and feasibility of the direct PCR assay for rapid diagnosis of acanthamoeba keratitis were evaluated and compared with culture method,corneal smear examination and real-time PCR.Results Direct PCR primers could only amplify DNA of acanthamoeba rather than other pathogens,and 10 stains of acanthamoeba were detected at least in each sample.During the development of acanthamoeba keratitis in the mice,the diagnosis positive rate of direct PCR was 80.0％,90.0％,80.0％,70.0％,70.0％ and 50.0％ in 1 day,3,5,7,10,15 days after modeled with the total positive rate 73.3％,which was higher than 31.7％ of culture method,56.7％ of corneal smear examination and 61.7％ of realtime PCR,with a significant difference between the direct PCR and culture method (P =0.005),but no significant difference was seen in the total positive rate between the direct PCR and real-time PC R (P =0.172) or corneal smear examination (P =0.056).Conclusions The direct PCR assay is a simple,rapid,highly specific and sensitive method for the rapid diagnosis of acanthamoeba keratitis,especially for the limited lesion sample.

Objectives:To explore the protective effects of continuous trickle of low temperature liquids through the lateral ventricle on neurons after rabbit local cerebral ischemia reperfusion. Methods: The middle cerebral artery (MCA) of New Zealand rabbit was clipped with micro aneurysm clip for 2 hours and reperfused for 24 hours. Immediately after clipping the MCA, we trickled the left lateral ventricle continuously with low temperature liquids to decrease the brain temperature to mild hypothermia(32-35℃) and maintained for 2 hours. After reperfusion for 24 hours, we assessed the animal's neural deficits, observe the pathology of the ischemic brain tissues dyed by HE and determined the dry wet ratio for brain edema in the sham operated group, the control group and the mild hypothermia group respectively.Results:①The grades of neural deficits in mild hypothermia group were obviously lower than that in the clipping group( P 0.05 );②The dry wet ratios were obviously different in the mild hypothermia group and clipping group;③ Pyknosis and dense dying by HE were observed in the neural nuclei of ischemic cortex tissues of the clipping group, but no obvious changes were observed in the mild hypothermia group. Conclusions:Trickling ventricle with low temperature liquids could alleviate cerebral ischemia reperfusion injury, ameliorated neural deficit after ischemia reperfusion and were protective on the brain.

Objective To explore the relationship between the osteonecrotic volume (lesion size), angle and the index of necrotic extent on the femoral head. Methods Fifty-one hips in 39 patients with non-traumatic avascular osteonecrosis on the femoral head were divided into 12 equal segments from the head to the neck junction (a turning point of the spherical curve of the head) with whole hip displacement, each with 30 degrees on a coronal plane of weight-bearing surface. The osteonecrotic angle of the arc of each necrotic segment from the center of the femoral head was measured at the point of one- to 12-o'clock on imaging of two dimensional reconstruction of computerized tomography (CT) scans and anteroposterior and lateral radiographs. Necrotic volume on each segment of the femoral head was calculated with fluid displacement method. The index of osteonecrotic extent on the femoral head was calculated using direct anatomical measurements. Results Osteoneerotic volume on the femoral head at the point of 12- to one-o'clock on coronal plane was (74. 5 ± 7.4)% of the sphere equivalent of the whole femoral head, which was positively correlated to its necrotic angle [ (41.9±8. 3) degrees] at the point of one-o'clock on the plane, with a coefficient of correlation of 0. 843, and that at the point of one- to two-o'clock on the plane was (73.7 ±0. 4) %, which was positively correlated to its necrotic angle [ (41.9 ± 1.8) degrees] at the point of two-o'clock, with a coefficient of correlation of 0. 543. Osteonecrotic volume on the point of 11- to 12-o'cleck was (83.6±8.6)%, and the necrotic angle at the point of 12-o'clock was (44. 9±3.9) degrees, which were positively correlated each other, with a coefficient of correlation of 0. 701 (P <0. 01 ). Osteonecrotic volume on the femoral head was positively correlated to its necrotic angle , modified index of necrotic extent, index of necrotic extent and Kerboul conjugated necrotic angle, with coefficients of correlation of 0. 798, 0. 701, 0. 377 and 0. 398 ( P < 0. 01 ), respectively, at the point of one o' clock. Conclusions Measurements of osteonecrotic volume was positively correlated to the index of necrotic extent and necrotic angle on the femoral head, respectively. Necrotic angle on the plane at 30-degree of the trochanter on the femoral head can well reflect its necrotic volume.

Objective To evalute the accuracy of stroke volume variation (SVV) in monitoring blood volume in patients undergoing off-pump coronary artery bypass grafting.Methods Twenty-one ASA Ⅱ or Ⅲ patients of both sexes aged 44-77 yr undergoing off-pump coronary artery bypass grafting were enrolled in this study.Anesthesia was induced with midazolam,etomidate,fentanyl,rocuronium and dolicaine and maintained with target-controlled infusion of propofol,infusion of remifentanil,intermittent iv injetion of atracurium and inhalation of sevoflurane.The patients were mechanically ventilated (VT 8 ml/kg,RR 12 bpm,I:E 1:2,PEEP 0,FiO2 80％ ).PEr CO2 was maintained at 35-44 mm Hg.Radial artery was cannulated and connected to FloTrac pressure transducer and Vigileo monitor.6％ hydroxyethyl starch 130/0.4 sodium chloride solution 7 ml/kg was infused at a rate of 0.25 ml· kg- 1 1· min- 1 at 5 min of haemodynamics stabilization after pericardiotomy (T1).HR,MAP,CVP,systemic vascular resistance (SVR),systemic vascular nesistance index (SVRI),SVV,stroke volume index (SVI)and CI were recorded at T1 and at 10 min after loading dose (T2).The change rate of HR(△HR),MAP(△MAP),CVP(△CVP),SVR(△SVR),SVV(△SVV),SVI(△SVI) and CI(△CI) were calculated.△SVI≥25％ was considered effective volume expansion.The ROC curves for HR,MAP,CVP,SVR and SVV in determining the volume expansion efficacy were plotted.The area under the curves and 95 ％ confidence interval were calculated.Results Compared with T1,CVP,SVI,CO and CI were significantly increased,SVRI and SVV decreased at T2 (P ＜ 0.01).There was no significant difference in MAP and HR between T1 and T2(P ＞0.05).△SVI was negatively correlated with △HR and △SVR ( r =- 0.737,r =- 0.480,P ＜ 0.05).△SVI was not correlated with △CVP,△MAP and( P ＞ 0.05).The change in SVI was determined by SVV 8.8％ (sensitivity =52.6％,specificity =100.0％ ).The area under the curve for SVV and 95％ confidence interval were 0.579(0.346-0.812).Conclusion SVV can not be used to accuratelymonitor the changes in blood volume in patients undergoing off-pump coronary artery bypass grafting.

Objective To investigate the diagnostic value of twinkling artifact in urinary calcium stones.Methods Calcium oxalate monohydrate,hydroxyapatite and whitlockite stone phantoms were prepared and embedded in porcine kidneys for ultrasound scanning.The length and width of twinkling artifact were measured.The intensity of twinkling artifact and the contrast-to-noise ratio (C/N ratio) of acoustic shadowing were recorded and all the data was analyzed statistically.Results All the stone phantoms generated twinkling artifact and acoustic shadowing.The difference of the intensity,length and width of twinkling artifact and the C/N ratio of acoustic shadowing had statistically significant each composition (P ＜0.01).There was no overlap between 95％ confidence interval of the intensity of twinkling artifact for any two compositions,and any two compositions of stones could be differentiated by the intensity of twinkling artifact (P ＜0.05).The C/N ratio of acoustic shadowing of calcium oxalate monohydrate and whitlockite stone phantoms were higher than those of hydroxyapatite ones (P ＜0.05).The receiver operating characteristic (ROC) curve displayed that the cutoff value of 1 127 of the intensity of twinkling artifact could be used to diagnose hydroxyapatite stones.The sensitivity,specificity,positive predictive value (PPV),negative predictive value (NPV) and area under the curve (AUC) were 74 ％,68 ％,69.8％,72.3％ and 0.743 respectively.Conclusions Twinkling artifact could be used to differentiate the calcium stones,which will be helpful for clinical treatment.

Objective To investigate the effect of maternal deprivation (MD) on neurobehavior and PP1Cγgene expression in hippocampus. Methods Male pups were randomly divided into MD group(thirty-five)and control group(twenty-four). From PND 1 to PND 21 ,pups in the MD groups underwent daily maternal deprivation for 3 h ( Postnatal day). Neurobehavior was observed to investigate neurodevelopment, Morris water maze was used to measure spatial learning and memory,and Real-Time quantitative PCR was employed to analyze PP1Cγ gene expression. Results Several significant deficiencies were observed in bodyweight and grasping reflex while a great enhancement in hot-plate test in rat pups suffering from MD( (26.23 ± 2.81 )g vs. (30. 38 ± 3.85 )g;( 19.37 ± 11.89) s vs. (22.39 ± 17.62 ) s; (4.36 ± 1.76 ) s vs. ( 5.26 ± 2.55 ) s; P ＜ 0. 05 ), but deficiencies in neurological reflexes were subtle ( ( 0.83 ±- 0.30 ) s vs. ( 0. 83 ± 0. 34 ) s; ( 3.68 ± 1.63 ) s vs. ( 5.61 ± 3. 01 ) s;( 3.00 ± 0.00 ) vs. ( 3.00 ± 0. 00); P ＞ 0. 05 ). MD had a subtle influence on spatial learning and memory (P ＞0.05). Meanwhile,MD could lead to PP1Cγ expression down-regulation on PND 22 ( (2.19 ±0.62) vs. (3.52 ±0.86), P＜0. 05)which was in line with early neurobehavior results. No difference was found compared with MD group and control group on PND60 ( ( 1.73 ± 0. 78 ) vs. ( 1.33 ± 0. 34); P ＞ 0.05 ). However, there was the up-regulation of PP1Cγexpression on PND 90 ( (2.85 ± 0. 34) vs. ( 1.34 ± 0.93 ); P ＜ 0.05 ). Conclusion MD alters early neurobehavior and hippocampal PP1Cγgene expression in the Wistar rats,but has a subtle effect on learning and memory. At the same time,MD can make PP1Cγexpression in the hippocampus varying with the age.

Objective To investigate the possible role of nerve growth factors (NGF) and its receptors in ovarian follicle development,and detect the difference between polycystic ovarian symdrome (PCOS) and normal control.Methods Immunohistochemisty was applied to detect the expression of NGF,p75NTR,TrkA in ovarian follicle granulosa cells.Results In 54 cases (including 9 cases of IVM-PCOS,16 cases of IVF-PCOS and 29 cases of IVF-Normal),49 cases showed NGF protein positive in granulose cells with significance difference between groups (P＜0.05),and especially NGF was the strongest expressed factor in IVF-PCOS group but there was no remarkable difference between the other two groups in the expression of NGF.41 were TrkA protein positive,almost all IVF cases were TrkA protein positive while only 2 in IVM and there was no remarkable difference in expression of TrkA between IVF PCOS and IVF Normal(P＞0.05).The cases with stronger expression of NGF or TrkA had higher estradiol level than that of lower expression group (P＜0.05).51 cases had positive expression of p75 NTR protein but there was no significant difference between groups (P＞0.05).Conclusions TrkA is expressed in mature ovarian follicle,and the interaction of NGF-TrkA may be involved in ovarian follicle development,and overexpression of NGF may be associated with PCOS development.