·AIM: Impairment of choroidal perfusion was found in AMD patients. We postulated that vasoactive agents,which can reduce choroidal blood flow resistance, might prevent the development of choroidal neovascularization (CNV). D-Timolol and L-Timolol are hypotensive agents used in cardiovascular and glaucoma therapy. Their effects on laser-induced experimental CNV rat model and human umbilical vein endothelial cells (HUVEC) were thus evaluated.·METHODS: Male Brown Norway rats were anesthetized to receive Nd:YAG laser to break the Bruch's membrane. D-Timolol and L-Timolol were given once daily through intraperitoneal injection after laser treatment for 4wk. Fluorescein angiography was performed on 2wk and 4wk. HUVEC were tested by proliferation assay and adhesion assay with D-Timolol and L-Timolol at different concentrations.· RESULTS: D-Timolol reduced the fluorescein leakage to 83% of the control group in laser-induced rat's CNV model at a dosage of 15mg/(kg·d). L-Timolol had no effect on CNV formation even at a higher dosage of 20mg/(kg·d). D-Timolol inhibited the endothelial cells proliferation significantly by 300mg/L. L-Timolol also significantly inhibited the cell proliferation at 1 000mg/L. But at a lower dose such as 300mg/L, no significant inhibitory effect was found. Both drugs showed no effect on cell adhesion function in cell culture experiments.· CONCLUSION: D-Timolol was found to prevent CNV development in laser-induced model in vivo and inhibit vascular endothelial cells proliferation in vitro. L-Timolol had no effect on cell proliferation at the same dose, and neither on rat CNV model. The results indicate these two isomers have different functions on rat's CNV prevention and on HUVEC cell proliferation.

Objective To evaluate the use of PET/CT in the diagnosis of breast cancer. Methods In this study,33 patients with suspicious breast tumor underwent PET/CT imaging. The images of the breast were analyzed for qualitative assessment of increased tracer uptake and blood perfusion with PET/CT. Results Among 27 cases with pathology proved breast cancer,25 was judged as PET/CT positive,2 was false-negative. Sensitivity, specificity and accuracy of PET/CT in identifying breast cancer were 92.6%,100%,93.9%. Conclusion PET/CT is a reliable and sensitive measure in the diagnosis of breast cancer in vivo.

Objective　To explore the effect of nordihydroguaiaretic acid (NDGA) on the expressions of vascular endothelial growth factor (VEGF) and its receptor, kinase-inserted domain containing receptor(KDR) and the possible mechanism. Methods　The expression of VEGF in human malignant glioma cell line SHG-44 and that of KDR in human umbilical vein endothelial cell (HUVEC) line ECV-304 were observed 1～3 d after NDGA treatment with immunohistochemistry, in situ hybridization and image analysis. Results　The expression of VEGF was declined at protein or mRNA levels in SHG-44 cells after treated with 100 μmol/L NDGA for 1 to 3 d. The expression of KDR in endothelial cells with 100 μmol/L NDGA treatment for 1 to 3 d was decreased too, in a more obvious way compared with the decline of VEGF expression in SHG-44 cells. Conclusion　The results suggest that NDGA inhibits the expression of VEGF in glioma cells as well as that of VEGF receptor KDR in endothelial cells, which may be the important molecular mechanism of anti-angiogenesis of NDGA.

0.05). Conclusions In comparison with mammography,PET/CT has a higher degree of sensitivity and specificily in detecting breast cancer,and higher positive predictive value.PET/CT can provide more aspects of in vivo diagnostic information which may be useful in selecting therapeutic strategy and may supplement the inadequacies of mammography.

Allo-cell transplant rejection is associated with class II major histocompatibility complex (MHC II), while its transactivator (namely C II TA) regulates MHC II molecules expression strictly and exclusively. The aim of this study was to investigate the inhibiting effect of C II TA anti-sense RNA on MHC II expression. The cDNA for anti-sense RNA recognizing the 114-523 sequence of C II TA (arC II TA) was obtained from Raji cell by RT-PCR, and then inserted into the pcDNA3.1B plasmid (pcDNA3.1B-arC II TA, pD-arC II TA). Raji cells were transfected stably with pD-arC II TA, classic MHC II antigen (HLA-DR, -DP, -DQ) expression was assayed by flow cytometry (FCM). mRNA abundance of C II TA, invariant chain and classic MHC II were detected by RT-PCR. The results showed that compared with control (sense C II TA), the expression inhibition of HLA-DR, -DP, -DQ on pD-arC II TA positive Raji cell was 65.93%, 54.14%, 68.32% respectively. The mRNA contents of C II TA, invariant chain and classic MHC II also decreased. In conclusion, arC II TA inhibited C II TA and thus the family of MHC II molecules were regulated by it, therefore these results provide a novel approach for the control of graft versus host diseases.
Cell Line, Tumor ; Flow Cytometry ; Graft Rejection ; genetics ; prevention & control ; HLA-DP Antigens ; biosynthesis ; genetics ; HLA-DR Antigens ; biosynthesis ; genetics ; Humans ; Nuclear Proteins ; genetics ; RNA, Antisense ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; genetics ; Transfection

Objective To investigate the clinical features and causes of misdiagnosis of complex partial status epilepticus (CPSE) in the elderly. Methods The clinical data of elderly patients with CPSE admitted in our department between January and December, 2008 with previous misdiagnosis were reviewed. The diagnosis of CPSE was established according to the diagnostic criteria of the International League Against Epilepsy (ILAE). All the patients underwent video-EEG examination, head magnetic resonance imaging (MRI), and routine biochemical examination, and were followed up for at least 3 months. Results Seven female CPSE patients were involved in this analysis including 2 with frontal lobe epilepsy and 5 with temporal lobe epilepsy. The conditions were misdiagnosed as a variety of diseases including Alzheimer's disease, cerebral infarction, coronary heart disease, encephalitis, hysterism, and schizophrenia, and the duration of the misdiagnoses ranged from 2 days to 40 years. Video-EEG showed slow background, continuous interictal discharges or high-voltage slow waves in the temporal or frontal lobe, or ictal discharges of complex partial seizures. After treatment with antiepileptic medications, 3 patients were free of epileptic episodes within 3 months, 3 still remained in complex partial status, and 1 had repeated CPSE. Conclusions Complex symptoms, atypical EEG, co-morbidity and social factors all contribute to misdiagnosis of CPSE in the elderly. Suspected patients should undergo video-EEG examination for a definite diagnosis as early as possible.

Objective To study the clinical features and genetic mechanism of myoclonic-astafic epilepsy (MAE) in infancy. Methods This study was conducted among 10 infants with MAE (including 7 male and 3 female patients) diagnosed between 2006 and 2008 according to the criteria of International League Against Epilepsy (2001). The clinical data including onset age, seizure type, physical signs, EEG, brain maguetic resonance imaging (MRI), effects of anti-epileptic drugs and prognosis were analyzed. The mutations of voltage-gated sodium channel subunit type 1 gene (SCN1A gene) were screened by denaturing high performance liquid chromatography and direct sequencing. Results The 10 MAE cases included 8 sporadic cases and 2 with a family history of febrile seizure and epilepsy. The onset age ranged from 5 months to 39 months, and all the MAE patients had multiple generalized seizure types, including myoclonic-atonic, myoclonic, atonic, tonic-clonic and absence seizures. Two patients had myoclonic status epilepticus, and 7 showed mental retardation. All the patients showed normal findings in MRI. SCN1A gene was screened in 8 of the MAE patients, and no mutation was found. Valproate, clonazepam and levetiracetam were effective in these MAE cases. Conclusion MAE is a rare epilepsy syndrome, whose genetic mechanism is still unclear. Valproate, clonazepam and levetiracetam are effective for MAE, which is associated with poor prognosis.

OBJECTIVETo establish the norms of Sub-Health Measurement Scale Version1.0 (SHMS V1.0) for Chinese civil servants.

METHODSWe sampled a total of 15 000 civil servants form Tianjin (north China), Guangdong (south China), Anhui and Hunan (central China), Xinjiang (northwest China) and Shenyang (northeast China) to perform the spot trial, and established the mean, percentile and threshold norms based on the characteristics of SHMS V1.0 scores for Chinese civil servants.

RESULTSThe established norms based on the average scores of SHMS V1.0 showed a mean score of 66.55∓12.36 for young male subjects (below 40 years), 67.42∓12.40 for older male subjects, 66.22∓11.81 for female subjects younger than 40 years, and 65.94∓11.91 for older female subjects. The threshold norms of SHMS V1.0 divided 5 health states, namely disease, severe sub-health, moderate sub-health, mild sub-health and healthy states according to the Mean∓SD and Mean∓0.5SD of the converted scores. The 4 cut-off points were close to the 15th, 30th, 70th and 85th percentile scores of SHMS V1.0.

CONCLUSIONWe have established SHMS V1.0 norms for Chinese civil servants, which facilitates further investigation of the incidence of sub-health state and its contributing factors in civil servants.

Adult ; China ; Female ; Health Knowledge, Attitudes, Practice ; Health Promotion ; methods ; Health Status ; Health Status Indicators ; Humans ; Male ; Middle Aged ; Reference Values ; Surveys and Questionnaires

The phyA(m) gene encoding acid phytase and optimized neutral phytase phyCs gene were inserted into expression vector pPIC9K in correct orientation and transformed into Pichia pastoris in order to expand the pH profile of phytase and decrease the cost of production. The fusion phytase phyA(m)-phyCs gene was successfully overexpressed in P. pastoris as an active and extracellular phytase. The yield of total extracellular fusion phytase activity is (25.4+/-0.53) U/ml at the flask scale and (159.1+/-2.92) U/ml for high cell-density fermentation, respectively. Purified fusion phytase exhibits an optimal temperature at 55 degrees C and an optimal pH at 5.5~6.0 and its relative activity remains at a relatively high level of above 70% in the range of pH 2.0 to 7.0. About 51% to 63% of its original activity remains after incubation at 75 degrees C to 95 degrees C for 10 min. Due to heavy glycosylation, the expressed fusion phytase shows a broad and diffuse band in SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). After deglycosylation by endoglycosidase H (EndoH(f)), the enzyme has an apparent molecular size of 95 kDa. The characterization of the fusion phytase was compared with those of phyCs and phyA(m).
6-Phytase ; genetics ; isolation & purification ; metabolism ; Amino Acid Sequence ; Fermentation ; Genetic Vectors ; Molecular Sequence Data ; Pichia ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; isolation & purification

OBJECTIVETo report a case of pulmonary surfactant protein (SP) gene mutation associated with pediatric interstitial lung disease, and study the clinical diagnosis process and review of related literature, to understand the relationship between interstitial lung disease and SP gene mutation in infants and children.

METHODThe clinical, radiological, histological, and genetic testing information of a case of SP gene mutation related pediatric interstitial lung disease were analyzed and related literature was reviewed.

RESULTA 2-year-old girl without a history of serious illness was hospitalized because of the shortness of breath, cough, excessive sputum, and the progressive dyspnea. Physical examination on admission revealed tachypnea, slight cyanosis, and the retraction signs were positive, respiratory rate of 60 times/minute, fine crackles could be heard through the lower lobe of both lungs; heart rate was 132 beats/minute. No other abnormalities were noted, no clubbing was found. Laboratory test results: pathologic examination was negative, multiple blood gas analysis suggested hypoxemia. Chest CT showed ground-glass like opacity, diffused patchy infiltration. Bronchoalveolar lavage fluid had a large number of neutrophils, and a few tissue cells. Eosinophil staining: negative. Fluconazole and methylprednisolone were given after admission, pulmonary symptoms and signs did not improve, reexamination showed no change in chest CT. Then lung biopsy was carried out through thoracoscopy. Histopathology suggested chronic interstitial pneumonia with fibrosis. The heterozygous mutation of R219W in the SFPTA1 and the S186N in SFTPC were identified by SP-related gene sequencing. The review of related literature showed that polymorphisms at the 219th amino acid in SP-A1 allele were found in adults with idiopathic pulmonary fibrosis (IPF), but there is no related literature in pediatric cases. The patient in this report had a mutation at the SP-A1 allele consistent with related literature. Data of 17 young children with mutation in SP-C gene showed that all the 17 cases had dyspnea and tachypnea, chest CT revealed diffuse opacities in lungs, the pathology of lungs was NSIP and CPI. There were 17 kinds of mutation and the common mutation was I73T. The mutation of S186N in SFTPC in our case has never been shown in previously published literature.

CONCLUSIONA case of interstitial lung disease with S186N gene mutation in SFTPC was preliminarily diagnosed in an infant. The SP-C gene mutations and polymorphisms are associated with pediatric interstitial lung disease.

Biopsy ; Child, Preschool ; DNA Mutational Analysis ; Dyspnea ; diagnosis ; pathology ; Female ; Humans ; Infant ; Lung ; diagnostic imaging ; pathology ; Lung Diseases, Interstitial ; diagnosis ; genetics ; pathology ; Male ; Mutation ; Pulmonary Surfactant-Associated Protein C ; genetics ; Tomography, X-Ray Computed