2.Expression, purification and antibody preparation of alpha-hemolysin from Staphylo-coccus aureus
Di LI ; Yaping GAO ; Peng JIN ; Jie DONG ; Yu LIU ; Guang YANG
Military Medical Sciences 2014;(11):871-874,884
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.
3.A distributed storage architecture for regional medical image sharing and cooperation based on HDFS.
Peng-jun LI ; Guang-jie CHEN ; Wen-ming GUO
Journal of Southern Medical University 2011;31(3):495-498
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
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Information Storage and Retrieval
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Radiology Information Systems
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Software
4.Repair of canine segmental mandibular defects by using autogenous bone marrow stromal cells and?-tricalcium phosphate
Jie YUAN ; Lian ZHU ; Guang-Peng LIU ; Feng XU ; Yu-Lai WONG ; Lei CUI ; Wei LIU ; Yi-lin CAO
Chinese Journal of Trauma 2003;0(09):-
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.
5.Prospective study of first night effect on 2-night polysomnographic parameters in adult Chinese snorers with suspected obstructive sleep apnea hypopnea syndrome.
Jing MA ; Cheng ZHANG ; Jue ZHANG ; Jie HU ; Jing FANG ; Jie ZHANG ; Peng JIA ; Guang-Fa WANG
Chinese Medical Journal 2011;124(24):4127-4131
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
6.DNA extraction from old bones.
Jie ZHANG ; Qing-jie XUE ; Guang-feng TANG ; Lei PENG
Journal of Forensic Medicine 2006;22(1):72-73
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*
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Cadaver
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DNA/isolation & purification*
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DNA Fingerprinting
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Femur/chemistry*
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Forensic Anthropology/methods*
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Humans
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Polymerase Chain Reaction/methods*
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Sensitivity and Specificity
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Specimen Handling/methods*
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Tandem Repeat Sequences
7.FANCA gene mutation analysis in Fanconi anemia patients.
Fei CHEN ; Guang-Jie PENG ; Kejian ZHANG ; Qun HU ; Liu-Qing ZHANG ; Ai-Guo LIU
Chinese Journal of Hematology 2005;26(10):616-618
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
8.Coronary anomaly: the single coronary artery.
Xu-Guang QIN ; Wei-Guo XIONG ; Chun-Peng LU ; Cheng-Jie GONG ; Li-Hua SHANG
Chinese Medical Journal 2010;123(7):972-973
9.Effect of polygoni multiflori radix preparata on diminished ovarian reserve in rats
Can ZHU ; Yao-Feng LI ; Fang PENG ; Chang-Fu YANG ; Guang-Qin CHEN ; Ting LU ; Jie LIU
Journal of Medical Postgraduates 2018;31(6):602-607
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.
10.Clinical efficacy of desmopressin in the treatment of mild hemophilia A in children.
Song-Ting BAI ; Jie LU ; Guang-Yao SHENG ; Song-Tao XU ; Lei XIE ; Shao PENG
Chinese Journal of Contemporary Pediatrics 2011;13(9):715-717
OBJECTIVETo study the effects of desmopressin (DDAVP) on coagulation factor Ⅷ (FⅧ) and activated partial thromboplastin time (APTT) in children with mild hemophilia A.
METHODSEighteen children with mild hemophilia A were enrolled. DDAVP (0.3 μg/kg•d) was injected intravenously for 5 days. Plasma FⅧ levels and APTT were measured before and after DDAVP treatment.
RESULTSIn 16 of 18 children with mild hemophilia A, the bleeding symptoms, including the articular or musclar hematoma, were significantly alleviated as a result of DDAVP treatment. The plasma FⅧ levels increased significantly to (27±4)% and APTT was shortened to (66±10)s 60 minutes after the first dose of DDAVP treatment. The plasma FⅧ remained at the levels of 25%-30% during 3-4 days of DDAVP treatment. Five days after DDAVP treatment, the plasma FⅧ levels decreased [(21±3)%], and APTT was prolonged when compared with 1-4 days of DDAVP treatment.
CONCLUSIONSDDAVP treatment can increase plasma FⅧ levels and shorten APTT in children with mild hemophilia A. DDAVP is effective in the treatment of mild hemophilia A. The duration of DDAVP therapy for mild hemophilia A is recommended as 3 to 4 days.
Child ; Child, Preschool ; Deamino Arginine Vasopressin ; therapeutic use ; Factor VIII ; analysis ; Hemophilia A ; blood ; drug therapy ; Humans ; Infant ; Male ; Partial Thromboplastin Time