In this study, toluidine blue O stain was conducted in the lung smears of rats and mice with Pneumocystis carinii infection induced by corticosteroids. All of the cyst wall of Pneumocystis carinii was stained dark purple and was easily distinguished from the blue background. The parenthesis-like structure in the cyst could clearly be recognized in the lungs with both specimens just as with the Grocott's methenamine-silver nitrate method. It is considered that this structure can be a distinctive mark from fungi. The results of the experiment show that the toluidine blue O stain may become a practical technic for the diagnosis of Pneumocystis carinii pneumonia and can substitute the Grocott's method.

Objective To investigate the PCR-based evaluation of prednisolone-induced relapse of asymptomatic Toxoplasma gondii infection and the therapeutic efficacy of azithromycin.Methods A total of 36 of female ICR mice,about 20 g,were randomly divided into 6 groups:contrast group (C),prednisolone group (P),infection group(I),infection plus prednisolone group (IP),infection plus azithromycin group(IA),infection plus prednisolone and azithromycin group (IPA).The infection group (I),infection plus prednisolone group(IP),infection plus azithromycin group(IA),infection plus prednisolone and azithromycin group (IPA)were challenged at week 0 with 10 cysts of Toxoplasma gondii Prugniaud strain per injection intraperitoneally.The prcdnisolone group (P),infection plus prednisolone group (IP) infection plus prednisolone and azithromycin group (IPA)were injectied with prednisolone 1 mg into hind medial subcutaneous every day from the 6th week to 7th week.The infection plus azithromycin group(IA),infection plus prednisolone and azithromycin group (IPA) were injectied with azithromycin 250 mg/kg intraperitoneally every day from the 6th week to 7th week.The serum samples were collected and DNAs extracted at week 0,1,2,3,4,5,6 and 7 for amplification of Toxoplasma gondii of specific B1 gene by PCR.All the mice were sacrificed 7 weeks after the challenge to calculate the number of cysts in brain tissues.Results Compared with the primer of AF146527 gene,the primer of B1 gene was more sensitive and specific.The method of PCR could amplify the productions of specific B1 gene Toxoplasma gondii 5 weeks before the challenge,while it could not amplified 5 weeks after the challenge.All the mice of the IP group were dead 2 weeks after the injection of prednisolone (week 7),and the only two mice of the IPA group were dead at the same time (P <0.05),respectively.Compared with the I group,IA group and IPZ group,the number of cysts in brain tissues of the IP group significantly increased (P <0.01).Conclusions B1 as target gene is more suitable for diagnosis of Toxoplasma gondii infection by PCR.Prednisolone could induce the relapse of asymptomatic Toxoplasma gondii infection of mice and the mice are finally dead.Azithromycin is effective but it can not completely cure the Toxoplasma gondii infection.

Objective To study the efficiacy and safety of transfacet approach decompression to treat thoracic spinal stenosis caused by anterior compression.Methods Thirty-three patients with thoracic spinal stenosis caused by anterior compression were treated in our institution from April 2005 to April 2009.Nineteen patients with more than 12 months follow-up were included in this study.Among of them,10 were male and 9 were female,with the age ranged from 33 to 77 years(mean,55.9 years).The causes of compression for spinal stenosis included ossification of posterior longitudinal ligament(OPLL)in 5 cases,thoracic disc herniation(TDH)in 11 cases,OPLL with ossification of ligamentum flavum(OLF)in 2 cases and TDH with OLF in 1 case.All patients underwent anterior decompression via a transfacet approach combined with anterior fusion and posterior fixation.The modified Japanese Orthopaedic Association(JOA)score system and Nurick Myelopathy grade were used to evaluate the outcomes.Results The operation time ranged from 180 to 480 min,with an average of 299.5 min and the blood loss was varied from 250 to 2200 ml,with an average of 918.5 ml.Among 7 cases with OPLL(including combined with OLF),2 patients developed neurologic deterioration and 1 patient developed cerebrospinal fluid leakage.There were no neurologic deterioration,cerebrospinal fluid leakage and other complications occurred in 12 cases with TDH(including combined with OLF).The follow-up ranged from 12 to 54 months(mean,28.6 months).The preoperative JOA score ranged from 2 to 11(mean,6.3).The JOA score in the last follow-up ranged from 5 to 11(mean,8.6).According to Nurick Myelopathy grade,the preoperative grade was 0 in 2 cases,1 in 2,2 in 4,3 in 5,4 in 2,and 5 in 4.The number of postoperative grade was 6,6,3,3,1 and 0 respectively.Conclusion Satisfactory decompression could be achieved by using transfacet approach for thoracic spinal stenosis caused by anterior compression.The approach is a safe and promising alternative for thoracic spinal anterior decompression.

Objective To evaluate carotid endarterectomy for patients with atherosclerotic occlusive diseases. Method Surgical experience of endarterectomy in 32 patients with atherosclerotic occlusive diseases was retrospectively analyzed. Internal carotid artery stem pressure was measured during operation. Shunting was used routinely. Patching was used electively. Anti-platelet agents and agents reducing plasma fibrinogen were given perioperatively. Result There was no perioperative mortality, nor ischemic cerebral stroke. Patients were followed-up from 5 months to 20 months with no occurrence of ischemic cerebral stroke during the follow-up. One case presented dysfunction of hypoglossal nerve. Two cases presented dilation of carotid artery on ultrasound scan. There were no restenosis (≥50%) and thrombosis. Conclusion Carotid endarterectomy for patients with atherosclerotic occlusive diseases of carotid artery is effective and safe. Careful performance, shunting, patching and the correct order of clamps removing are the keys of preventing complications.

Objective To explore the diagnosis and management of ruptured abdominal aortic aneurysm(RAAA).Methods Twelve patients with RAAA treated in past 7 years were revienled retrospectively.The main clinical manifestations were abdominal pain and / or back pain,low blood pressure or shock,and pulsating abdominal mass.All cases were accurately diagnosed with CT and 7 were treated by conventional operation,one by EVAR,and the other 4 did not receive surgical treatment.Results Perioperative death occurred in 5 cases(mortality rate was 62.5%) in 8 surgical treated patients,including circulatory failure in 2 cases,renal failure in 1 case,and multiple organ failure in 2 cases.All the 4 patients treated with nonoperative method were dead.Conclusions Surgical operation in RAAA cases still carried a high mortality.Early dignosis,appropriate resuscitation,urgent surgical repair,reduction of operative time,and infrarenal clamping are measures conducive to lowering the mortality rate of RAAA.EVAR has the potential to reduce the mortality rate from RAAA.

The ultrastructural changes of atrial myocardial cells during hemorrhagic shock and after autotransfusion with the shedded blood were observed in rats,which were divided into control,hemorrhagic shock and auspeto-transfusion groups.It was found that there was a marked increase of atrial specific granules in the atrial myocardial cells during hemorrhagic shock.Striking ultrastructural changes of atrial myocardial cells were noted after transfusion of the shedded blood.Reduction in the number of atrial specific granules,in addition to mitochondrial swelling,reduction of the cristae,focal emptying of mitochondria,myelin-like figure in mitochondrial matrix,expansion of sar-coplasmic reticulum,dilatation of Golgi complexes and perinuclear space,and dissociation of myocardial disk,were remarkable.The findings suggest that atrial myocardila cells participate in the patho-physiological changes of hemorrhagic shock and autotransfusion.

Objective To study the effect of sevoflurane and propofol anesthetic techniques on interleukin (IL)-6 and IL-10 in patients with laparoscopic hysterectomy.Methods Fifty elective laparoscopic hysterectomy patients were randomly divided into sevoflurane group (25 patients) and propofol group (25 patients) who received either sevoflurane or propofol for their anesthesia.After induction,adjusted the sevoflurane inhalation concentration in sevoflurane group and propofol pumping speed in propofol group.Maintained the Bispectral index (BIS) value at 50 +5.Recorded heart rate (HR),mean arterial blood pressure (MAP),BIS,IL-6,IL-10 on 5 min before anesthesia (T1),10 min after pneumoperitoneum (T2),40 min after pneumoperitoneum (T3) and 5 ain before the end of the operation (T4),and compared.Results The level of BIS,HR,MAP in two groups and between two groups had no significant difference (P ＞ 0.05).The level of IL-6,IL-10 on T2-T4 were significantly higher than those on T1 [sevoflurane group:(31.0 ± 9.0),(33.0 ± 11.0),(34.0 ± 16.0) ng/L vs.(29.0 ± 8.0) ng/L and (19.3 ± 1.7),(24.0 ± 2.8),(27.0 ± 8.0) ng/L vs.(2.0 + 0.4) ng/L; propofol group:(38.0 ± 9.0),(40.0 + 12.0),(45.0 ± 18.0) ng/L vs.(29.0 + 11.0) ng/L and (8.2 ± 2.3),(11.0 ± 4.2),(18.0 ± 7.0) ng/L vs.(2.0 ± 0.3) ng/L] (P ＜ 0.05).The level of IL-6,IL-10 on T1 between two groups had no significant difference (P ＞ 0.05).The level of IL-6 on T2-T4 in sevoflurane group was significantly lower than that in propofol group and the level of IL-10 on T2-T4 in sevoflurane group was significantly higher than that in propofol group (P＜ 0.05).Conclusions At maintaining the balance of cytokines in laparoscopic hysterectomy,the effect of sevoflurane is better than propofol.Sevoflurane is more suitable for maintenance of anesthesia for laparoscopic gynecologic operation.

Objective To express human cytotoxic T lymphocyte associated antigen-4 (CTLA4) extracelluar domain in Bacmid-baculovirus expression system. Methods The cDNA of human CTLA4 extracelluar domain was isolated from plasmid pUC19-CTLA4Ig by PCR.The specific cDNA was subcloned into plasmid pFastBacl. The insertion was confirmed by DNA sequencing, and then transposed into Hz8 Bacmid in DH10B. The recombinant Bacmid was used to transform Hz insect cells. The supernatant and cellular lysate were analyzed by using SDS-PAGE. Results A specific protein in the cellular lysate of Hz insect cells containing human CTLA4 extracelluar domain with MW38 x 103 was found. Conclusion Human CTLA4 extracellular domain is expressed in the Bacmid-baculovirus expression system.

Objective To investigate the correlation of spinal canal stricture and intramedullary increased signal intensity (ISI) in patients with ossification of the posterior longitudinal ligament (OPLL). Methods 92 patients with OPLL were divided into 3 groups, those with the sagittal diameter remained ≥66.7% were as group A, 33.3%~66.7% as group B, and <33.3% as group C. The incidence of intramedullary ISI was recorded, and their neurological condition was assessed with the Japanese Orthopedics Association Assessment (JOA). Results ISI were found in 6 cases in the group A (20.7%), 17 cases in the group B (47.2%) and 19 cases in the group C (70.4%) (P<0.05). The score of JOA was (7.1±2.1) in the group A, (6.0±1.8) in the group B and (5.6±2.0) in the group C (P<0.05). Conclusion The incidence of intramedullary ISI increased with the severity of spinal canal stricture, and with more severe nerve damage in OPLL patients.

Objective To study the probability of applying mesenchymal stem cells isolated from human umbilical cord blood (hUCB MSCs) to repair mouse skin wound in vivo. Methods hUCB MSCs isolated from full term delivery human umbilical cord blood were cultured and amplified in vitro.hUCB MSCs at passage 9 were labeled with BrdU (5-bromodeoxy-uridine) and grafted on the full-thickness skin loss wound created on the back of the severe combined immunodeficiency (SCID) mouse (treatment group), when a PBS control group was set. The wound healing rate was surveyed and compared at days 7 and 14 postoperatively. Meanwhile, the wound was biopsied at days 7, 14 and 28 after operation,and the expressions of BrdU antibody and K19 antibody were checked pathologically and immunohistochemically by HE staining, respectively. Results The wound in treatment group was healed more rapidly than that in control group (P ＜ 0.01 ). The pathological check of the biopsy sample showed that the epidermis was thicker, with more epidermal ridges in the treatment group, compared with control group.It was found that some BrdU positive cells were distributed successively on the hair follicle, the stratum basal and the spinosum layers, a few of which even expressed K19. Conclusion hUCB MSCs can be differentiated into skin tissue and cells and is possible to repair skin wound.