AIM:To study the KIF21A gene mutation in a Han family with concomitant exotropia. ·METHODS: The genomic DNA of five family members was extracted from peripheral blood leukocytes and amplified with PCR. The PCR products were purified for DNA sequencing. DNA sequences were aligned with the human KIF21A gene sequences registered in GenBank. · RESULTS: Mutation analysis of all exons of the pedigree's KIF21A gene reveals no gene mutation in any of the families. ·CONCLUSION: Our study demonstrates that the KIF21A gene maybe is not virulence gene in this pedigree.

Canrenone is an important intermediate for the synthesis of eplerenone,a cardiovascular drug.C_ 11 ?-hydroxylation of canrenone is the key reaction,which can be done by microbial transformation.Rhizopus sp.SIPI-0602,kept in our lab,could high selectively transform canrenone to a compound named SIPI-11.By determining and analyzing the MS,UV,NMR etc.spectra of compound SIPI-11,its chemical structure was elucidated to be 11?-hydroxycanrenone.The study on flask transformation technology showed that the transformation ratio exceeded 90% when the substrate concentration was not more than 6g/L.

OBJECTIVETo investigate the role of perindopril for prevention and treatment of glucocorticoid-induced osteoporosis (GIOP) in rabbits.

METHODSA total of 45 male New Zealand white rabbits (10 months old, weight 3.0 to 3.5 kg) were randomly divided into 3 groups involving normal control group (muscle injection of saline solution, n = 15, group NC), model group (muscle injection of dexamethasone, n = 15, group GIOP), and treatment group (muscle injection of dexamethasone combined with oral perindopril, n = 15, group GIOP+ACEI). All rabbits put to death after 12 weeks' treatment. The changes of bone mass and strength were observed and analyzed by bone histomorphology, biomechanics, metabolic bone related serological indexes and mRNA expression.

RESULTSAt 12 weeks, the analysis of bone histomorphology and biomechanics results showed that the bone mass and bone strength of group GIOP were significantly lower than that of group NC (P < 0.05); after perindopril treatment, the bone mass and bone strength of group GIOP+ACEI were higher obviously than that of group GIOP (P < 0.05). Mineralizing surface,mineral apposition rate and serum osteocalcin in group GIOP decreased than group NC; however, osteoclast number, osteoclast surface, eroded surface, and urinary deoxypyridinoline in group GIOP increased than group NC (P < 0.05); these changes were inhibited after perindopril treatment (P < 0.05). Quantitative RT-PCR revealed that after dexamethasone treatment, the ratio of SOST mRNS expression and RANKL/OPG mRNA expression obviously increased than that of group NC (P < 0.05); and Runx2 expression decreased significantly (P < 0.05); while the changes of mRNA expression were improved by perindopril treatment.

CONCLUSIONPerindopril can promote bone formation and inhibit bone resorption to deduce glucocorticoid-induced osteoporosis. This study provides a new method for prevention and treatment of GIOP.

Animals ; Biomechanical Phenomena ; Glucocorticoids ; adverse effects ; Male ; Osteoporosis ; chemically induced ; prevention & control ; Perindopril ; therapeutic use ; Rabbits

AIM:To explore the association of the mutation in PPP2R3A exons and retinoblastoma.METHODS:Hospital-based case control study was taken.Retinoblastoma patients (15 cases, as case group) and matched controls (30 controls, as control group) were recruited in this study.Genomic DNA obtained from formalin fixed paraffin embedded (FFPE) and peripheral blood were used as template.PPP2R3A gene exon sequences were detected by PCR-sequencing.Homology analysis was performed using blastn in GenBank.RESULTS:Analyzing PPP2R3A DNA sequences (1001bp) from 15 cases, two reported SNPs had been detected, including rs34629706 and rs144802055.Rs34629706 also occurred in the control group.Rs144802055 appeared only in the case group.CONCLUSION:PPP2R3A gene SNPs of rs34629706 is unrelated to the incidence of retinoblastoma.Relations between rs144802055 and RB needs to be further explored.

Objective To retrospectively evaluate the mammographic features of breast ductal carcinoma in situ (DCIS)and DCIS with small invasive foci,and to analyze the correlation between the mammographic findings and the prognostic biologic factors.Methods The mammographic examination was performed in 95 consecutive women with breast DCIS(n = 50)and DCIS with invasive foci(n = 45 ).The prognostic biologic factors including progesterone receptor(PR),C-erbB-2,and p53 were evaluated in 62 of 95 cases.Categorical data were expressed as percentages and analyzed by using the X~2 test,and furthermore the odds ratio was measured.Results(1)Only one abnormality was seen on mammography in 62 patients. Combined two abnormalities on mammography were seen in 26 patients.Mammograms were normal in 7 patients.(2)Calcifications with or without other abnormality were noted in 62 cases.Of them,73% (n =45)had higher probability of malignancy calcifications and the others were intermediate concern calcifications.Clustered calcifications(36 lesions)was the most common distribution,which usually accompanied by another abnormality.And then were segmental(18 lesions)distributed pattern.As far as the shape of mass (n = 22)was concerned,the oval shaped lesion(13 cases)was the most common,and the margin of the mass appeared as ill-defined in 15 eases,microlobulated in 1,circumscribed in 4,and obscured in 2,respectively.Isodensity mass had a higher frequency in this group(12/22,55%).Other non-calcification findings included architecture distortion(7 cases),local asymmetry (15 cases),global asymmetry (5 cases),and solitary dilated duct (3 cases),and most of them accompanied with other signs. (3)For expression profile of the biological factors,significant differences were found among malignant calcification group,intermediate concern calcification group,and non-calcification group. The odds of PR positive for the lesions noted as non-calcification were 11.00 times higher (X~2 =8.571 ,P=0.003 ;95% CI, 1.998—60.572)than the lesions noted as intermediate concern calcifications,and 8.80 times higher (X~2 = 9.748,P=0.002 ;95% CI,2.024—38.253)than the lesions noted as malignant calcifications.The odds of C-erbB-2 positive for the lesions showed as malignant calcifications were 12.35 times higher (X~2=7.353, P=0.007 ;95% CI,1.447—105.443)than the lesions showed as non-calcification,and 5.74 times higher (X~2=4.977,P = 0.026;95% CI,1.110—29.645)than the lesions showed as intermediate concern calcifications.Conclusion The mammographic features of DCIS and DCIS with small invasive foci were characteristic.Mammographic findings could be a prognostic markers,which could provide a possibility for making a treatment plan.

Objective：This study was designed to investigate the apoptosis of HL 60 cells induced by the peptide from hemocyte of horseshoe crab (horseshoe crab hemocyte peptide). Methods： Cytotoxicity and cell viability were assayed by MTT method. Morphologic assessment of apoptosis was performed with fluorescence microscope; the apoptosis of the cells was confirmed by flow cytometry; cell cycle was analyzed by flow cytometry, changes of cell membrane were observed by scanning electron microscope. Results： HL 60 cells treated with horseshoe crab hemocyte peptide were showed apparently cytotoxicity with IC50 24 μ g/ml. The morphologic changes including reduction in volume, nuclear chromatin condensation, fluorescence strength were observed with fluorescence microscope. Treated with horseshoe crab hemocyte peptide 50- 100 μ g/ml for 6 h, apoptostic HL 60 cells were predominant. However the HL 60 cells were shown necrotic and apoptostic cells were reduced when treatment with horseshoe crab hemocyte peptide of 200 μ g/ml. SubG1 peak was detected by flow cytometry. Cell cycle analysis showed Phase G1 cells decreased and Phase G2 cells increased with horseshoe crab hemocyte peptide 50 μ g/ml treating for 0- 12 h. Ratio of apoptosis increase depended on concentrations of horseshoe crab hemocyte peptide in 25- 100 μ g/ml but it reduced in 200 μ g/ml, the HL 60 cells were shown necrotic mainly. Scanning electron microscope showed membrane of HL 60 cells was damaged by horseshoe crab hemocyte peptide treatment,cell membrane showed some holes. Conclusion： Peptide from hemocytes of horseshoe crab can induce apoptosis in HL 60 cell,it was early event,and may correlate with membrane damage. The HL 60 cell in Phase G1 seemed to be sensitive to the peptide.

A new cadinane-type sesquiterpenoid, pogocablene P (1), and a new natural product with cyclohexanone skeleton, pogocablone A (2), were isolated from the EtOAc soluble fraction of the aerial parts of Pogostemon cablin by several chromatographic methods, such as silica gel, Sephadex LH-20, ODS and high performance liquid chromatography (HPLC), and so on. Their structures were identified by means of mass spectrometry and nuclear magnetic resonance spectroscopy. In addition, the absolute configuration of compound 2 was determined by electronic circular dichroism (ECD) calculation. Furthermore, the anti-influenza virus and anti-inflammatory activities of compounds 1 and 2 were evaluated.

Objective To study the anti-fatigue effect of Cynomorium songaricum flavone(CSF)on old rats.Methods50 old male Wistar rats were randomly divided into the control group(no swimming,no drugs),single swimming group(swimming,given saline by gavage),and three swimming plus CSF groups(swimming,given CSF 20 g/kg weight,10 g/kg weight and 5 g/kg weight respectively),total five groups with 10 animals in each group.Animals swam with free swimming in endless cycles water 30 minutes once every day for 10 days.The level of monoamine oxidase(MAO) and glutathione peroxidase(GSH-px) in blood of animals were examined by MAO kit,GSH-px kit and ultraviolet spectrophotometer at 11th day.ResultsCompared with single swimming group,the body weights of swimming plus CSF groups increased significantly by 12.9%,80.6% and 200% respectively(P<0.01);swimming times before sinking increased by 19.4%,29% and 48.4%;sinking times of each rat(time/10 min) decreased by 17.8%,43.2% and 72.9%;total swimming times increased by 2.8%,7% and 29.6% respectively(P<0.01);MAO decreased by 60%,69.8% and 80.5%,GSH-px increased by 465.3%,563.8% and 635% respectively(P<0.01).ConclusionCSF shows anti-oxidative and anti-fatigue properties and can be given as prophylactic/therapeutic supplements for increasing antioxidant enzyme activities and preventing lipid peroxidation during strenuous exercise.

Objective:To use polyamidoamine(PAMAM)dendrimer as gene delivery system for survivin gene anti- sense oligodeoxynucleotide(asODN)transfection for inhibition of HepG2 cancer cell growth.Methods:The first to the fifth generation of PAMAM and asODN were used to prepare a complex:PAMAM-asODN.The morphology of PAMAM- asODN was observed using agrose electrophoresis and atomic force microscope(AFM).PAMAM-asODN was then used to transfect HepG2 cells and cells transfected with asODN served as control.The transfection efficacy of PAMAM-asODN into HepG2 cells was observed under confocol microscope,the surviving mRNA expression was analyzed by RT-PCR,and the inhibition of HepG2 cell growth was determined by MTT assay.Results:Agrose electrophoresis showed strong complexing action between PAMAM and asODN and they formed a complex with a diameter of 25 nm.Confocol microscope showed the transfection efficacy of PAMAM-asODN was higher than that of asODN.RT-PCR showed a decreased expression of sur- vivin mRNA in PAMAM-asODN transfected cells.MTF results demonstrated that the growth of HepG2 cell was obviously inhibited after transfection of PAMAM-asODN and the inhibition rate increased with culture time,concentration of com- plex,the generation of PAMAM.PAMAM-asODN at 6.0?mol/L G4.0 resulted in a 55% inhibition of HepG2 cells 96 h after culture.Conclusion:PAMAM dendrimers can efficiently mediate the entry of survivin asODN into HepG2 cells,re- sulting in inhibition of HepG2 cells.PAMAM might be a promising gene carrier for potential molecular therapy of cancer.

Objective To study the clinical features and prevention means of the infection caused by the highly dathogenic avian influenza A(H_5N_1).Method The clinical data which were confirmed to be an H_5N_1 infected case were analyzed retrospectively.Results The patient,16-year-old male,had no unambi guous history of direct contact with diseased or dead poultry before the onset of the disease.After the onset of the disease,chest X-ray showed flake shadow of the right lower lung quickly spread to the whole lung,associated with mediastinum,subcutaneous emphysema,acute respiratory distress syndrome(ARDS)was occurred on the fifth day.Mechanical ventilation is the primary measure in the comprehensive treatment.On the sixth day,Chinese Center for Disease Control and Prevention detected the pharyngeal specimen of the patient by RT - PCR,Real- time PCR method and suggested positive for the A/H5/N1 virus nucleic acid andis01ated the avian flu(H_5N_1) virus.The disease course was 10 days from onset of illness to death.Conclusions Human infection by the highly pathogenic avian influenza A(H_5N_1)is a deadly infectious disease.If the lesions is widespread and associated with ARDS,prognosis is poor.