Objective A prokaryotic expression vector was constructed by inserting the coding sequence of mouse sperm specific lactate dehydrogenase C into pET-28a(+)and the recombinant mLDHC44 protein was purified by Ni+-NTA agrose.Methods The cDNA of mouse sperm specific lactate dehydrogenase C was obtained by RT-PCR,with total RNA of mouse testis tissues as templates.The coding sequence of mouse LDHC4 was amplified by PCR with specific primers.This recombinant vector was transformed into Escherichia coli BL21(DE3).The recombinant mLDHC4 protein was induced by isopropy-?-D-thiogalactoside(IPTG)and identified by sodium dodecyl sulfate polyacrylamide electrophoresis and LDH activity determination.After purified with Ni+-NTA agrose,the mLDHC4 protein was probed with antisera from the pVAX1-mLDHC4 vaccine(the eukaryotic expression vector of mouse sperm specific lactate dehydrogenase C)immunized BALB/c mice by Western blot analysis.Results After digested with BamH I-EcoR I,the recombinant plasmids produced right fragment which was about 1000bp.Sequencing showed that the sequence of the cloned fragment was in agreement with sequence in GenBank.This recombinant vector was named as pET-28a(+)-mLDHC4.With induction of IPTG,The recombinant protein with molecular weight of about 35 kD was expressed and the enzyme activity of this protein was high.After purified with Ni+-NTA agrose,this mLDHC4 protein formed a specific band by sodium dodecyl sulfate polyacrylamide electrophoresis and probed with antisera from immunized BALB/c mice and then formed a specific band in the nitrocellulose membrane.Conclusion The coding sequence of mouse lactate dehydrogenase subunit C had been cloned into the prokaryotic expression vector pET-28a(+)and the mLDHC4 protein could be expressed at a high level,the specificity of this protein was high and the activity was strong.

With the rapid development of pharmacogenetics, more and more studies have shown evidence in the association between polymorphisms at the human leukocyte antigen (HLA) loci and severe adverse drug reactions (SADRs). Several HLA-B alleles proved to be associated with SADRs for drugs such as carbamazepine, allopurinol, lamotrigine, and lfucloxacillin. hTe USA Food and Drug Administration (FDA) has even recommended routine screening for HLA-B allele before the use of abacavir and carbamazepine. With the completion of human genome project and the Hapmapproject, several new pharmacogenetics approaches such as genome-wide association study (GWAS) have emerged. hTese newly developed methods will undoubtedly accelerate the identiifcation and clinical utilization of the pharmacogenetic biomakers. In addition, the immunogenetic mechanisms by which the HLA alleles cause SADRs are explored at the cellular and molecular level. hTis review focuses on the recent progresses in HLA alleles and ADRs regarding both the clinical translation and modern pharmacogenetic methods.

Objective To study the correlation between benign prostatic hyperplasia (BPH) and vesicoureteral reflux (VUR). Methods Intravenous radionuclide cystography (IVRC) was performed on 88 BPH patients and micturition cystourethrography on 30 to find if there was any VUR and the degree of reflux. Results 31 patients had VUR of which there were 6 grade Ⅰ,7 grade Ⅱ,6 grade Ⅲ,6 grade Ⅳ and 6 grade Ⅴ.9 were on the right,6 on the left and 16 bilateral. Conclusions VUR do occur in some BPH patients and IVRC is the better method for its diagnosis and evaluation.

Objective By compared FAB classification of myelodysplastic syndrome with WHO classification of myelodysplastic syndrome,understanded the proposed WHO new classification of myelodysplastic syndrome as soon as possible.Methods Retraced and analysed 78 cases of myelodysplastic syndrome based on examination of the bone marrow.Restults(1)According to the FAB classification,divided myelodysplastic syndrome into 5 kinds:RA,RARS,RAEB,CMML,RAEB-T.(2)According to the WHO classification,its subtypes were adjusted again,and its concluded:RA,RARS,RCMD,RAEB-Ⅰ,RAEB-Ⅱ,5q-del.(3)There were 9 cases possess chromosome to revise particularly in the chromosome check 28 cases myelodysplastic syndrome. Conclusion WHO classification has clinical guiding significance for early diagnosis,therapy observation and prognosis decision.

Objective To summary the color Doppler imaging feature of localized Castleman disease.Methods From January 1997 to November 2011,32 localized Castleman diseases which were proved by pathology were analyzed.Results Round-like,hypoechoic,hypervascular lesions were showed in 23 hyalinevascular type lesions,2 of them with calcium,3 of them with structure liked lymph node hilum.Round-like,hypoechoic,hypervascular lesions or normal lymph node were showed in 5 plasma- type lesions and 4 mixed type lesions.Conclusions When round like,hypoechoic,hypervascular lesion is found by ultrasonography,Castleman should be considered.Calcium or lymph nod hilum-like structure is special finding in diagnosis of Castleman disease by ultrasonography.

AIM:To observe the effects of prostaglandin E2 on transforming growth factor-?1(TGF-?1)triggered human lung fetus fibroblast(HLF)transdifferentiation and connective tissue growth factor(CTGF),collagen type I(COLⅠ)expression.METHODS:HLFs were treated with TGF-?1,the cells underwent phenotypic change to myofibroblast.The marker of myofibroblast-?-smooth muscle actin(?-SMA)was detected by immunofluorescence.The ?-SMA content was measured by Western blotting.The changes in CTGF and COL Ⅰ at transcription levels were estimated by RT-PCR method.CTGF protein expression was evaluated by immunocytochemical.Cell culture medium hydroxyproline amount was measured by colormetric assay.RESULTS:PGE2 blocked TGF-?1 induced ?-SMA positive myofibroblast transformation(P

A new enhancement method is proposed based on the characteristics of fundus images in this paper. Firstly, top-hat transform is utilized to weaken the background. Secondly, contrast limited adaptive histogram equalization (CLAHE) is performed to improve the uneven illumination. Finally, two-dimensional matched filters are designed to further enhance the contrast between blood vessels and background. The algorithm was tested in DIARETDB0 databases and showed good applicability for both normal and pathological fundus images. A new no-reference image quality assessment method was used to evaluate the enhancement methods objectively. The results demonstrated that the proposed method could effectively weaken the background, increase contrast, enhance details in the fundus images and improve the image quality greatly.
Algorithms ; Contrast Media ; Fundus Oculi ; Humans ; Image Enhancement

Objective To investigate the dynamic changes of platelets (PLT) and white blood cells (WBC) after traumatic brain injury (TBI) and discuss its clinical significance. Methods The number of PLT and WBC were examined in 63 patients with TBI by using cytoanalyze and also analyzed together with Glasgow Outcome Scale and concurrent infection, in the meantime, enzyme-linked immu-nosorbent was used to investigate concentration changes of C reactive protein (CRP) and thrombospondin 1 (TSPI) and analyze the correlation between CRP and TSP1. Results The number of WBC in all pa-tients, whether concurred with infection or not, was significantly increased within 24 hours after TBI (P < 0.01), with no statistical difference between patients without infection at day 4 and normal patients (P >0.05). However, the number of WBC was decreased to below 10 × 109/L in patients without infec-tion, which was significantly higher than that in normal patients (P < 0.05). In patients with infection and unfavorable prognosis, the number of WBC was increased again ay days 7-14, whereas that of PLT rose significantly at days 14-21 (P <0. 01). The concentration of TSPI was positively correlated with that of CRP (r = 0.720, P < 0.01). Conclusions Monitoring the dynamic changes of PLT and WBC is promising. The change of WBC at day 4 post injury is a key indicator to provide evidences of prophylactic antibiotic usage. Much attention should be paid to the dynamic change of PLT at days 14-21 post injury so as to evaluate the condition of hypercoagulability that can be potentially caused by inflammation response. Secondary increase of WBC and later increase, of PLT may affect prognosis of the patients. TSP1 and CRP may participate in thrombosis formation induced by inflammation.

BACKGROUND: Previous studies have confirmed that rabbit bone marrow mesenchymal stem cells (BMSCs) can differentiate into osteoblasts under osteogenic induction in vitro, stably express the specific phenotype of osteoblasts and have osteogenic ability. Calf cortical bone scaffold with partial cancellous bone has good biocompatibility and degradability, which can be used as a carrier material of bone marrow mesenchymal stem cells. OBJECTIVE: To combine rabbit BMSCs with calf bone composite according to the basic principles of bone tissue engineering and to observe the osteogenesis in the New Zealand white rabbits after implantation of BMSCs/calf bone composite into the ilium, thereby providing a direct evidence for preliminary clinical application of tissue-engineered bone products.METHODS: BMSCs/calf cortical bone scaffold with partial cancellous bone (tissue-engineered bone group), simple calf heterogeneous bone (heterogeneous bone group) or autologous iliac bone (autologous iliac bone group) was randomly implanted into the rabbit ilium. The changes of implant surface and tissue reactions around the implant were observed.X-ray examination was performed to observe osteogenic changes at 4, 8, 12, 24 weeks after implantation. Immunohistochemistry staining was used to observe the expression of bone morphogenetic protein 2.RESULTS AND CONCLUSION: After heterogeneous bone implantation, the wound healed well, and there were no systemic or local inflammation and toxicity reactions in all groups. The X-ray results showed that at postoperative 24 weeks, the implant was basically fused with the host bone in the tissue-engineered bone group, but the fusion was unsatisfactory in the heterogeneous bone group. The process of ossifications from cartilages was observed in all groups by hematoxylin-eosin staining, and bone morphogenetic protein 2 was positive for immunohistochemical staining. Findings from in vivo experiments indicate that rabbit BMSCs seeded onto the calf cortical bone scaffold with partial cancellous bone could construct tissue-engineered bone by osteoinductation in vitro in the rabbits.

Objective: To analyze the clinical characteristics of young patients with acute myocardial infarction (AMI), discuss the key points of health education in young patients with AMI, enhance the understanding of patients, and improve the prognosis of AMI in the young patients. Methods: hTe patients were chosen in XiangyaHospital from September 2012 to September 2013. We consulted the medical records, analyzed the clinical characteristics and results of coronary angiogram in young patients (age≤45), and compared with old patients (age≥60). Results: There were 69 young patients with AMI, about 14.2% of all the patients with AMI. Of the 69 young patients, 59 were male (85.5%) and 10 were female (14.5%). Compared with the old patients, the percentages of smoking, drinking, hyperlipidemia and overweight were much higher;the percentages of hypertension and diabetes were much lower in young patients. The coronary angiogram showed that the constituent ratios of insigniifcant disease and single-vessel disease inthe young patients were higher than those in the old patients; the constituent ratios of double-vessel disease and triple-vessel disease in the young patients were lower than those in the old patients. Conclusion: The clinical characteristics of young patients withAMI are different from the old patients.Health education should be conducted in the youth, and new diet and lifestyle should be advocated.