Animals ; Autophagy ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Rats ; Rats, Sprague-Dawley

Objective To clone and express alpha hemolysin ( Hla ) , one important virulence factor secreted by Staphylococcus aureus in Escherichia coli to tdetect the hemolytic activity of Hla on erythrocytes from diffrerent species ,and to prepare specific antibodies against Hla that can inhibit the hemolytic activity of Hla .Methods Hla gene was amplified by PCR using S.aureus NCTC-8325 genome DNA as a template.The expression vector pET-28a-Hla was constructed and transformed into E.coli BL21(DE3).The recombinant Hla protein was expressed by IPTG induction ,and then purified by Ni2+affinity chromatography .The hemolytic activity of Hla was measured by erythrocyte lysis assays .Results Soluble recombinant Hla protein was expressed and purified .Further investigations showed that Hla could significantly induce the lysis of rabbit erythrocytes .However , erythrocytes from humans or sheep were more resistant to the lysis mediated by Hla . The specific polyclonal antibodies against Hla ( anti-Hla) were prepared and detected by ELISA .Moreover, it was found that anti-Hla could inhibit the hemolytic activity of Hla .Conclusion We found that Hla from S.aureus has different hemolytic effects on erythrocytes from various species .The prepared Hla-antibodies can specifically block the hemolytic activity of Hla.

Objective To repair segmental mandibular defects with autogenous bone marrow stromal cells(BMSCs)and?-triealcium phosphate.Methods Isolated BMSCs were in vitro expand- ed.A 3 cm-long segmental mandibular defect was created at right mandible in 12 canines,of which de- fects in six canines were repaired with BMSCs and?-tricalcium phosphate(?-TCP)and that in other six cases repaired with?-TCP,which was used as control.The engineered bone was evaluated by X-ray, CT,DXA,gross and histological examination,immunohistochemistry and biomechanical test 4,12,26,32 weeks after operation respectively.Results In induced BMSCs,histochemistry showed AKP activity. Oral X-ray showed obvious callus formation 4-26 weeks after operation in experimental group but minimal bone formation in control group.At 32 weeks after operation,gross observation,X-ray and CT demonstra- ted well bony-union in experimental group but bony-nonunion in control group.DXA indicated that the bone density of experimental group was significantly higher than that of control group.Biomechanical test revealed no statistical difference upon mechanical strength of mandibula between experimental group and normal group.Conclusions Canine segmental mandibular defects can be well repaired with the tissue- engineered bone generated by autogenous osteogenic BMSCs and?-TCP scaffold.

Given the importance of regional centers for medical image sharing and cooperation is important for resource balancing, healthcare service enhancement and medical expense reduction, building such regional medical image sharing and cooperation centers faces huge challenges. In this paper we analyze the advantages and weakness of two storage architectures, and designed a hybrid storage architecture combining FC SAN and Hadoop HDFS. A HDFS suitable medical image file format, called S-DICOM, and a set of S-DICOM operating middleware, SDFO (S-DICOM File Operator), was developed. The results of performance testing indicated that this hybrid storage architecture is suitable for storing and managing large volume of medical images.
Image Processing, Computer-Assisted ; Information Storage and Retrieval ; Radiology Information Systems ; Software

BACKGROUNDA prospective 2-night polysomnographic (PSG) study in Chinese snorers was designed to assess the role of the first night effect (FNE) in PSG parameters and the diagnosis of obstructive sleep apnea hypopnea syndrome (OSAHS).

METHODSSeventy-two snorers from two teaching hospitals underwent overnight PSG on two consecutive nights. The night-to-night variability of PSG parameters were recorded and analyzed.

RESULTSSixty-six patients were analyzed. Among all the PSG parameters, only the total time of stage 2 presented a significant difference between two nights: 219.50 (83.50 - 353.50) vs. 215.25 (59.50 - 342.50) (P = 0.000). Subgroup assessment showed a slight night-to-night difference in about 1 - 2 parameters in the group with the apnea-hypopnea index (AHI) ≥ 20 events per hour as well as the group with AHI < 20 events per hour, but there was no night-to-night difference in the AHI in each group. And slighter FNE was found among patients ≥ 40 years old. There was no significant difference in diagnosis of OSAHS. In the decision of severity, a slight difference was found between the two nights with a Kappa value = 0.531.

CONCLUSIONSOnly mild FNE can be found on two consecutive nights of PSG in adult Chinese snorers, but it has no effect on the diagnosis of OSAHS. A single polysomnographic night may be adequate for the diagnosis of OSAHS.

Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Polysomnography ; methods ; Prospective Studies ; Sleep Apnea, Obstructive ; physiopathology ; Snoring ; physiopathology ; Young Adult

OBJECTIVE: To explore an efficient method for extracting DNA from old bones. METHODS: Using an organic combined with and Microcon 100 to extract and purify DNA. RESULTS: The extracted DNA was successfully genotyped by using florescence labeling STR multiplex amplification. CONCLUSION This method will be useful for forensic scientists in identification of DNA from old bones.
Bone and Bones/chemistry* ; Cadaver ; DNA/isolation & purification* ; DNA Fingerprinting ; Femur/chemistry* ; Forensic Anthropology/methods* ; Humans ; Polymerase Chain Reaction/methods* ; Sensitivity and Specificity ; Specimen Handling/methods* ; Tandem Repeat Sequences

Aged ; Angioplasty, Balloon, Coronary ; Coronary Angiography ; Coronary Artery Disease ; diagnosis ; therapy ; Coronary Vessel Anomalies ; diagnosis ; therapy ; Female ; Humans

OBJECTIVETo screen the FANCA gene mutation and explore the FANCA protein function in Fanconi anemia (FA) patients.

METHODSFANCA protein expression and its interaction with FANCF were analyzed using Western blot and immunoprecipitation in 3 cases of FA-A. Genomic DNA was used for MLPA analysis followed by sequencing.

RESULTSFANCA protein was undetectable and FANCA and FANCF protein interaction was impaired in these 3 cases of FA-A. Each case of FA-A contained biallelic pathogenic mutations in FANCA gene.

CONCLUSIONSNo functional FANCA protein was found in these 3 cases of FA-A, and intragenic deletion, frame shift and splice site mutation were the major pathogenic mutations found in FANCA gene.

Cell Line ; DNA Mutational Analysis ; Fanconi Anemia ; genetics ; metabolism ; Fanconi Anemia Complementation Group A Protein ; genetics ; metabolism ; Humans ; Mutation

Objective Diminished ovarian reserve (DOR) severely affects the life of women and the estrogen replacement therapy for it has obvious adverse effects. This article aimed to study the effect of polygoni multiflori radix preparata (PMRP) on DOR in rats and provide a therapeutic option for clinical medication. Methods Sixty female SD rats were randomly divided into six groups of equal number,normal control,DOR model control,high-dose PMRP (4 g/kg),medium-dose PMRP (2 g/kg),low-dose PMRP (1 g/kg),and positive control. The DOR model was established by gavage of tripterygium glycosides as 75 mg/kg every morning,followed by administration of PMRP in the PMRP groups,Estradiol valerate at 0.18 mg/kg in the positive control group and distilled water in the model control group in the afternoon,all for 30 consecutive days. The estrous cycle of the rats was observed,the levels of serum estradiol (E2),follicle-stimulating hormone (FSH),luteinizing hor-mone (LH),anti-Müllerian hormone (AMH) and inhibin-B (INH- B) were determined by ELISA,the ovarian and uterine indexes were obtained,and the ovarian morphology was observed by HE stai-ning,and the counts of follicles at different stages were recorded. Results Compared with the normal controls,the DOR model rats showed modeling time-related lengthening,irregularity and even disorder of the estrous cycle,with a few epithelial cells or keratino-cytes and leucocytes on the vaginal smear at 11-30 days. The estrous cycle was normal in the PMRP and positive control groups at 1-10 days and relatively prolonged at 11-30 days. In comparison with the normal control group,the DOR model rats exhibited a signifi-cantly decreased levels of serum E2 ([302.6±42.9] vs [155.7±46.8] pg/mL,P<0.05) and INH-B ([494.5±84.1] vs [299.2± 106.8] pg/mL,P<0.05) but increased levels of FSH ([7.2±0.5] vs [21.7±1.2] mIU/mL,P<0.05) and LH ([17.4±1.2] vs [25.0±1.0] mU/mL,P<0.05). The INH-B level was markedly elevated in the PMRP and positive control groups as compared with that in the DOR models (P<0.05). The counts of follicles and corpora lutea were remarkably lower in the DOR model rats (P<0.05) while that of developing follicles markedly higher in the PMRP and positive control groups than in the normal control group (P<0.05). The numbers of atretic follicles+corpora lutea were significantly increased in the high-dose PMRP group but decreased in the low-dose PMRP group (P<0.05) and positive controls (P<0.05). The counts of primordial and developing follicles were dramatically higher in the PMRP and positive control groups than in the DOR model controls (P<0.01) and so were the numbers of atretic follicles+corpora lutea in the high-and medium-dose PMRP groups (P<0.05). Conclusion Polygoni multiflori radix preparata can effectively protect the reproductive function of female rats by inhibiting tripterygium glycosides-induced toxicity to the ovary.

OBJECTIVETo detect the level of transforming growth factor-beta1 (TGF-beta1), TGF-beta2, vascular endothelial growth factor (VEGF) and platelet-derived growth factor receptor-alpha (PDGFRalpha) in plasma and peripheral blood leukocytes in a hereditary hemorrhagic telangiectasia type 2 (HHT-2) family, and explore the implication of angiogenesis related proteins in HHT-2 pathogenesis.

METHODSThe diagnosis of the HHT-2 patient was based on clinical features and further confirmed by determining a C1231T mutation of activin receptor-like kinase 1 (ALK1) gene. Five other new members in this family were evaluated with ALK1 gene screening and clinical manifestation. Plasma level of TGF-beta1, TGF-beta2 or VEGF was measured by ELISA, and the expression of PDGFRalpha,TGF-beta1, and VEGF in peripheral blood leukocytes by flow cytometry combined with direct or indirect immunofluorescence.

RESULTSNo C1231T mutation was detected in exon 8 of ALK1 gene in the 5 new members. Plasma TGF-beta1 and TGF-beta2 concentration in 3 affected HHT case was (16 954 +/- 3 709) ng/L and (11 548 +/- 2 611) ng/L, respectively, compared with that of normal control, the difference was not significant (P > 0.05). VEGF concentration in the 3 HHT patients, 6 unaffected family members and 6 normal controls was (179.2 +/- 22.0) microg/L, (149.8 +/- 22.7) microg/L and (132.9 +/- 21.0) microg/ L, respectively. Plasma VEGF level in HHT patients was significantly higher than that in normal subjects (P < 0.025). Peripheral leukocyte PDGFRalpha and VEGF in HHT patients and unaffected family members were markedly higher than that of normal control (P < 0.05 and P < 0.02), while TGF-beta1 distribution was similar in HHT patients and normal subjects.

CONCLUSIONCompared with normal controls there is no difference in plasma TGF-beta1 concentration on peripheral leukocytes of HHT patients. Plasma VEGF concentration or leukocytes VEGF expression in HHT is significantly higher than that of normal subjects. Leukocytes PDGFRalpha expression in HHT is significantly higher than that of normal control. These changes may be associated with a compensable mechanism in HHT.

Adolescent ; Adult ; Aged ; Child, Preschool ; Female ; Granulocytes ; metabolism ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Pedigree ; Receptor, Platelet-Derived Growth Factor alpha ; blood ; Telangiectasia, Hereditary Hemorrhagic ; blood ; Transforming Growth Factor beta ; blood ; Vascular Endothelial Growth Factor A ; blood