Objective To summarize the clinical characteristics and causes of ischemic cerebrovascular disease(ICD)in children.Methods A retrospective analysis was conducted on the clinical data of 53 cases with ICD from Feb.2002 to Jun.2008 at the department of neurology in Wuhan Children's Hospital.The self-designed questionnaire of children with ICD was used,whose items included patients' age,gender,personal history,clinical features,cerebrospinal fluid examination,neurological imaging,immunologic examination,metabolic examination,and so on.Results Of 53 children with ICD,30 cases(56.6%)were male,and 23 cases(43.4%)were female.Patients' age varied from 9 months to 12 years old,in which 45 cases(84.9%)were less than 6 years old.Patients from rural area(60.4%)were more than those from city(39.6%).Ratio of limb paralysis was 75.5%(40 cases)in first clinical symptomatology of children with ICD,including hemiplegia in 32 cases(60.4%),alternate hemiplegia in 5 cases(9.4%)and monoplegia in 3 cases(5.7%).Skull CT/MRI scan was performed to reveal 27 cases(50.9%)with basal ganglia region infarction and secondly 15 cases(28.3%)with multi-lobar infarction.Forty cases were found in abnormal cerebrovascular image by means of magnetic resonance angiography/digital subtraction angiography,in which middle cerebral artery and its branches were involved in 21 cases(52.5%).There were 41 cases(77.4%)of patients to be found with clear causes,of which 13 cases(24.5%)were of infections,8 cases(15.1%)of moyamoya disease,5 cases(9.4%)of cerebral vascular malformations,4 cases(7.5%)of head trauma.However,another 12 cases(22.6%)of patients had unknown etiology.Conclusions Children with ICD had characteristics themselves.The limb paralysis was mostly the first symptoms,and the middle cerebral artery and its branches lesions were the most common locations in children with ICD,and next the internal carotid artery involvement,anterior cerebral artery involvement,posterior cerebral artery involvement,cerebral vascular malformations,and so on.Their major cause was infection,followed by Moyamoya disease,cerebrovascular malformations and head trauma,and there were still some unknown causes.

0.05),but there were statistically difference within E15,E19,P2,P10(Pa

A glycomic method was used to screen the aberrantly ?1-6 fucosylated glycoproteins related to HCC metastasis and analyze the alteration of CK8 both in its expression level and its glycan parts associated with metastatic ability. Based on the approach, 2-DE coupled with lectin affinity blot, lectin affinity precipitation followed by MALDI-TOF-MS/MS, the lens culinaris agglutinin (LCA) affinity glycoprotein profiles from MHCC97-L and MHCC97-H cells, two higher metastatic HCC cell lines, were obtained, in which a differentially displayed protein spot was indicated when compared with Hep3B, in the region within 55～60 ku in molecular mass and 4～6 in isoelectric point. The identification result was CK8 by MALDI-TOF-MS/MS. To confirm the relation between increased core-fucosylation of CK8 and HCC metastasis, LCA affinity precipitation was used to extract the ?1-6 fucosylated glycoproteins, followed by Western blot. And it was found that CK8 was highly fucosylated in both MHCC97-L and MHCC97-H cells compared to Hep3B. Immunofluorescence analysis and Western blot were used to detect its intracellular localization and its protein expression levels, indicating that CK8 distributed in cytoplasm and increased protein expressions in MHCC97-L and MHCC97-H cell lines. And further lectin binding studies found that CK8 has a high affinity for Con A in both MHCC97-H and Hep3B cells, indicating that CK8 was a glycoprotein with high-mannose type N-glycans. But the amount of the lectin RCA-1 binding to CK8 was greater in MHCC97-H than Hep3B, suggesting that CK8 contained the increased terminal galactose residues ?-1, 4-linked to GlcNAc in MHCC97-H. All the results suggested that the increase of CK8 in its protein expression level, core-fucosylation and terminal gal ?1,4 GlcNAc disaccharides might be related to HCC metastatic ability.

Objective To investigate the treatment and mechanism of aminoguanidine in retina of diabetic of rats .Methods To-tal 60 rats were divided into control group(n=20) ,diabetic group(n=20) and aminoguanidine treatment group(n=20)which would be treated by aminoguanidine for 14 days .Then the eye tissue of rats were took after 14 days administration for pathological obser-vation(HE staining) ,and the induced nitric oxide synthase(iNOS) ,endothelial nitric oxide synthase(eNOS) ,nerve type of nitric ox-ide synthase(nNOS) level and the expression of differences content and expression were investigated by ELISA ,Western blot and PT-PCR .Results HE staining showed that retinal tissue defects decreased and neuronal cells of rats in aminoguanidine treatment group were increased and significant (P<0 .05) compared rats in diabetic group .The iNOS content and expression of rats in amin-oguanidine treatment group were lower than diabetic group by ELISA ,Western blot and PT-PCR ,it was significantly difference (P<0 .05) and without significant difference between the normal group and diabetic group (P> 0 .05) .Compared with diabetes group ,iNOS ,eNOS ,nNOS protein expression in the rat retina in aminoguanidine treatment group were reduced (P< 0 .05) ,and without significant difference between the normal group and aminoguanidine treatment group (P>0 .05) .The iNOS mRNA expres-sion was lower than that of eNOS mRNA and nNOS mRNA in aminoguanidine treatment group .Conclusion Aminoguanidine can improve retinal tissue of diabetic rats with lesions ,the pathways may be selectively inhibit inducible nitric oxide synthase activity of iNOS .

Objective To observe the dynamic changes of neuron-specific enolase(NSE)mRNA and protein in offspring rats brain tissue with bilirubin encephalopathy and explore the pathological mechanism and its diagnostic value on bilirubin encephalopathy.Methods Seven-day postnatal Wistar rats were used for study.One hundred and twenty rats were divided into 2 groups randomly(control group and experimental group),which were respectively subdivided into 6 groups(6,12,24,48,72,96 h).The rats in control group were intraperitoneally administered physiological saline 0.5 mL,the rats in experimental groups were intraperitoneally administered bilirubin(200 mg/kg).Reverse transcription-polymerase chain reaction was used to analyze the dynamic changes of NSE mRNA expression at 6,12,24,48,72 and 96 h in brain tissue of rats with bilirubin encephalopathy.Immunohistochemistry was used to evaluate NSE protein expression in hippo-campi,cerebral cortex,thalamic and pallidus at different times.Results The expression of NSE mRNA significantly decreased in brain tissue of rats with bilirubin encephalopathy from 6 h to 96 h compared with the control groups.The expression of NSE protein in hippocampi decreased in offspring rats with bilirubin encephalopathy from 6 h to 96 h,but there were no differences compared with the control groups.The expression of NSE protein in cerebral cortex was significantly decreased in rats with bilirubin encephalopathy from 6 h to 96 h,there were significant differences compared with the control group.The expression of NSE protein in thalamic significantly decreased in rats with bilirubin encephalopathy from 6 h to 96 h,but there were significant differences between experimental groups and the control groups at 24 h and 72 h.The expression of NSE protein in pallidus significantly decreased in offspring rats with bilirubin encephalopathy from 6 h to 96 h,and there were significant differences compared with control groups.Conclusions The changing trends of expression of NSE mRNA were identical to those of NSE protein.NSE may reflect the degree of injury of neurogliocyte.It can serve as reliable index to determine bilirubin encephalopathy.

OBJECTIVETo study the clinical pathologic and immunohistochemical features of gastrointestinal mesenchymal tumors (GIMTs), and to investigate the value of molecular markers in GIMTs clinical differentiation diagnosis.

METHODSThe clinical and pathological data of 210 cases of GIMTs, collected from Jan. 1987 to Dec. 2005 in our hospital, were investigated retrospectively. GIMTs were rediagnosed by using standard immunostaining technique in paraffin-embedded tissue. The expression level of CD117, CD34, Desmin, SMA and PS100 were detected by immunohistochemical method.

RESULTSAmong 210 cases of GIMTs, 127 cases were Gastrointestinal stromal tumors (GISTs) (60.5%), 33 leiomyomas and leiomyosarcomas (15.7%), 27 neurogenic tumours (12.8%), and 23 miscellaneous tumors (11.0%). The incidences of GIST, leiomyoma and leiomyosarcoma were similar among men and women. Men were more likely to develop neurogenic tumors and miscellaneous tumors than women. Of all the GISTs, 51.2% cases originated from stomach, 19.7% from small intestine, 11.0% from esophagus, 10.2% from colon and rectum. The most common location of leiomyomas and leiomyosarcomas was esophagus (45.5%). The most common location of neurogenic tumors was retroperitoneum (74.1%). Common symptoms of GISTs included digestive tract hemorrhage in 36 cases (28.3%), abdominal pain in 27 cases (21.3%) and abdominal mass in 24 cases (18.9%). Other GIMT cases except GISTs had no first symptom of digestive tract hemorrhage. It was noticed that 79.5% of GISTs had no obvious invasion, and 72.7% of leiomyomas and leiomyosarcomas had no obvious invasion. 33.3% of neurogenic tumors invaded the adjacent organs or tissues. No metastases had been found in other GIMT cases except GISTs. The neoplastic cells of GISTs were composed of various percentage of spindle (72.5%), epithelioid (11.8%) and mixed-type cells (15.7%). The percentage of spindle cells in leiomyomas and leiomyosarcomas was 94. The immunohistochemical results of GISTs showed that the positive rate of CD117 was 93.7%, CD34 was 69.3%, Desmin was 13.4%, SMA was 12.6%, and PS100 was 10.2%. The immunohistochemical results of leiomyomas and leiomyosarcomas showed that the positive rate of Desmin was 78.5%, SMA was 63.6%, while as the expressions of CD117, CD34, and PS100 were negative. Diffuse strong positive staining of PS100 was observed in 88.9% of neurogenic tumor patients.

CONCLUSIONSGISTs are the most common tumors among GIMTs. GISTs are different from neurogenic tumors, leiomyomas and leiomyosarcomas in initial symptom, tumor location, biological behavior and immunophenotype. Immunohistochemistry plays an important role in differentiating GISTs from leiomyomas and neurogenic tumors.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biomarkers ; analysis ; Child ; Child, Preschool ; Female ; Gastrointestinal Neoplasms ; pathology ; Humans ; Immunohistochemistry ; Infant ; Male ; Mesenchymoma ; pathology ; Middle Aged ; Young Adult

OBJECTIVETo study the clinicopathologic features of metastatic carcinoma in bone and to evaluate the role of immunohistochemistry in delineation of possible primary sites.

METHODSOne hundred and forty-one cases of metastatic carcinoma in bone encountered during the period from 1998 to 2004 in People's Hospital, Peking University, were reviewed retrospectively. The clinical information, radiographic features and pathologic findings were analyzed. Immunohistochemical study for antigens including cytokeratins, prostatic specific antigen, thyroglobulin, thyroid transcription factor 1 and gross cystic disease fluid protein 15, was performed in 51 cases possessing skeletal metastasis with unknown primary.

RESULTSSkeletal metastasis occurred more commonly in males (male to female ratio = 1.7:1). The age of patients ranged from 23 to 86 years (mean age = 56.5). The presenting symptoms included pain and dysfunction in the affected bones. The locations of skeletal metastasis were as follows: spine (58), pelvic bone (46), long bone (34) and others (3). Twenty-three cases harbored multiple bony lesions. Radiographically, 99 cases (70.2%) of skeletal metastasis were detected by X-rays, including 85 cases (85.9%) showing lytic changes. The primary sites of the tumor could be determined by clinicopathologic correlation in 90 cases (63.8%) and were unknown in the remaining 51 cases. Upon immunohistochemical study, the primary sites were determined in another 40 cases. Overall, the primary sites were identified in 130 cases (92.2%), which included lung (37), female genital system and breast (25), kidney (18), gastrointestinal system (17), liver (12), thyroid (11), prostate (7), bladder (2) and skin (1).

CONCLUSIONSSkeletal metastasis occurs more often in elderly males. Axial bones (spine and pelvis) are usually affected. Lung and female genital system are frequent the primary sites. Immunohistochemical study is useful in cases with occult primary.

Adult ; Aged ; Bone Neoplasms ; complications ; pathology ; Carcinoma ; complications ; pathology ; Female ; Humans ; Immunohistochemistry ; methods ; Male ; Middle Aged ; Neoplasm Metastasis ; pathology

Objective To observe the influences of uncoupling protein 2 (UCP-2) rs660339 variants transfection on cell proliferation and apoptosis of human umbilical vein endothelial cell (HUVEC). Methods Two UCP-2 green fluorescent protein (GFP) lentivirus constructs were created with the rs660339 locus carried C or T (UCP-2C or UCP-2T), respectively. HUVEC were cultured after lentiviral infection of UCP-2C or UCP-2T. The expression of UCP-2C or UCP-2T was detected with real time polymerase chain reaction. Cell proliferation and cell apoptosis were compared among negative control (NC) group, UCP-2T group and UCP-2C group using CCK-8 cell viability and flow cytometry. Western blot and immunostaining were employed to examine the expression of Bcl-2 gene. Results The lentivirus constructs were successfully created.>80%of the transfected cells were found to express GFP under fluorescent microscope. The mRNA levels of UCP-2 gene were significantly increased (F=29.183, P=0.001) in the UCP-2T group and UCP-2C group. The CCK-8 assay revealed that on day two (F=15.970, P=0.004), day three (F=16.738, P=0.004), day four (F=5.414, P=0.045) post-infection, UCP-2T and UCP-2C group showed significantly greater proliferation than the NC cells. The apoptotic rate in the UCP-2T and UCP-2C group was significantly lower than NC group (F=277.138, P=0.000), and the apoptotic rate of UCP-2T was significantly lower than that of UCP-2C (P=0.003). The protein levels of Bcl-2 in the UCP-2T and UCP-2C group were significantly greater than that in the NC group (F=425.679, P=0.000), and the Bcl-2 expression of UCP-2T was greater than that of UCP-2C (P=0.002). The Bcl-2 density in the UCP-2T and UCP-2C group were greater than that in the NC group (F=11.827, P=0.008), while there was no difference between UCP-2T and UCP-2C group (P=0.404). Conclusion The variants of UCP-2 rs660339 may influence HUVEC proliferation and apoptosis, and UCP-2T showed a stronger effect of inhibiting apoptosis than UCP-2C.

AIM:To explore the molecular mechanism of panaxadiol saponin(PDS)by observing Toll like receptor(TLR)2 and TLR9 mRNA expression induced by lipopolysaccharide(LPS).METHODS:Rats were divided into LPS,LPS+PDSL,LPS+PDSM and control group,respectively.Nitric oxide synthase(NOS)activity,nitric oxide(NO)content,LPO content,SOD activity and TLR2 and TLR9 mRNA expression were assayed 4 h after intravenous injection of LPS.RESULTS:NOS activity,NO content,LPO content of LPS+PDSL group and LPS+PDSM group were significantly lower than those in LPS group.TLR2 mRNA expression in the liver tissue of LPS+PDSL group and LPS+PDSM group was decreased compared with LPS group.CONCLUSION:PDS has a protective effect on liver tissues by triggering the down-regulation of TLR2 expression,reducing NOS activity,and NO content.

Osteonecrosis of the femoral head is frequently observed in patients treated with excessive corticosteroids. However, the pathogenesis of corticosteroid-induced osteonecrosis remains unclear. The purpose of this study was to investigate the role of Toll-like receptor 4 (TLR4) signaling pathway in steroid-induced femoral head osteonecrosis in rats. Male Sprague-Dawley rats were injected intramuscularly with 20 mg/kg methylprednisolone (MP) for 8 weeks, twice per week. The animals were sacrificed at 2, 4 and 8 weeks after the last MP injection, respectively, and then allocated to the 2-, 4- and 8-week model groups (n=24 each). Rats in the control group (n=12) were not given any treatment. Histopathological analysis was performed and the concentration of tartrate-resistant acid phosphatase (TRAP) in plasma was determined. The activation of osteoclasts in the femoral head was assessed by TRAP staining. The expression of TLR4, MyD88, TRAF6 and NF-κB p65 that are involved in TLR4 signaling, and MCP-1 production were detected by using real-time PCR (RT-PCR) and Western blotting. The results showed that the osteonecrosis in the femoral head was clearly observed and the concentration of TRAP in the plasma was increased in the model rats. The femoral head tissues in MP-treated rats were positive for TRAP and the intensity of TRAP staining was greater in MP-treated rats than in control rats. As compared with the control group, the mRNA expression of TLR4 signaling-related factors was enhanced significantly at 4 and 8 weeks, and the protein levels of these factors increased significantly with time. It was concluded that MP could induce the femoral head osteonecrosis in rats, which was associated with osteoclast activation via the TLR4 signaling pathway. These findings suggest that TLR4 signaling pathway plays a pivotal role in the pathogenesis of steroid-induced osteonecrosis.