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.

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.

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.

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.

According to different epochs and development needs, a series of practices on environmental health and sanitary engineering were carried out, which played significant roles in promoting national economic and social developments and protecting the public health. This paper reviewed the main achievements in the past 70 years infields of patriotic health campaign, water sanitation and toilet improvement in rural areas, surveillance and investigation, health standard system, sanitary engineering equipment, stove improvement etc., and then proposed several prospects in the future.

According to different epochs and development needs, a series of practices on environmental health and sanitary engineering were carried out, which played significant roles in promoting national economic and social developments and protecting the public health. This paper reviewed the main achievements in the past 70 years infields of patriotic health campaign, water sanitation and toilet improvement in rural areas, surveillance and investigation, health standard system, sanitary engineering equipment, stove improvement etc., and then proposed several prospects in the future.

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 experience of treating Budd-Chiari syndrome through orthotopic liver transplantation.Methods The clinical data of LTx performed on 9 patients with Budd-Chiari syndrome from December 2003 to April 2010 were retrospectively analyzed. We summarize the preoperative image and surgical experience,and observe the occurrence of postoperative complications and survival. Results Budd-Chiari syndrome was diagnosed in 9 patients by the preoperative abdominal CT enhancement and vascular reconstruction,and cavity venography was done to observe obstruction and sub-type of CAVA vein.All 9 patients were subjected to cadaveric liver transplantation.Eight cases accepted classic non bypass type,and one accepted living related right lobe liver transplantation. Postoperative triple immunosuppressive regimen included tacrolimus,mycophenolate mofetil,and hormone.The average follow-up periods for all these 9 patients were 32.8 months (13 to 61 months). One patient died from the tumor recurrence at 35th month after the operation.Two patients received re-transplantation for the lost of the graft.One recipient received the donor liver with medium steatosis,and the re-transplantation was performed on the12th day after the first transplantation due to the primary non function of the graft.The other one received the secondary liver transplantation at 6th month after the first transplantation due to the biliary complication and died from the liver tumor recurrence. Among all the 9 cases,seizure disorder (1 case),dysfunction of duodenal papillary muscle (1 case) and small-for-size syndrome (one case) occurred after the operation.Pulmonary infection occurred in 4 cases:3 cases due to the bacterial infection and 1 due to the fungal infection. Neither outflow obstruction nor the recurrence of the Budd-Chiari syndrome occurred in this study.The 1- and 2-year survival rate after the operation was both 100％,and 3-year survival rate post-transplantation was 88.9％ (8/9).Conclusion Liver transplantation can be the ideal treatment to the Budd-Chiari syndrome based on the definite clinical diagnosis,accurate imaging evaluation and eligible modus operandi.