In this study, nattokinase gene was amplified by PCR using bacillus subtilis chromosomal DNA as template and cloned into expressed vector pBV220. After transforming recombinant plasmid into E.coli HB101, the recombinant strain was yielded. It was proved that expression products was secretive and expression protein was 12% of total cell protein by SDS-PAGE. Optimum culture time and inducing time was determined as 6h and 5h respectively. The plasmid stability studies showed that recombinant plasmid has excellent segregational stability but the structural stability was not good in the host cell.

OBJECTIVETo investigate the correlation between power Doppler vascularity index (PDVI) and microvessel density (MVD) and evaluate the angiogenesis in high-grade gliomas and the adjacent edema in patients with glioma using intraoperative power Doppler ultrasound (PDUS) during gross total resection.

METHODSIn 25 cases of high-grade gliomas undergoing gross total tumor resections, PDUS was performed intraoperatively and the regions of interest within the tumor and the adjacent edema were analyzed with Photoshop software to measure the tumoral and peritumoral blood flow quantified as PDVI. The tumoral and adjacent MVD were determined using immunohistochemical staining for CD34. The correlation between PDVI in the gliomas and the adjacent edema and MVD in the corresponding areas were analyzed using Spearman correlation test.

RESULTSThe measurement of both PVDI and MVD revealed significant difference in vascularity between the gliomas and the adjacent edema (t=0.000, P<0.01), and PDVI was positively correlated to MVD measurement (r=0.7248 in the tumors and r=0.6608 in the adjacent edema).

CONCLUSIONSThe difference in the vascularity between the tumor and adjacent edema allows their distinction by PDUS during operation for high-grade glioma. Intraoperative PDUS provides an accurate and reliable means for measuring vascularity in the glioma and the adjacent edema tissue.

Adult ; Brain Edema ; diagnostic imaging ; etiology ; Brain Neoplasms ; blood supply ; complications ; diagnostic imaging ; Echocardiography, Doppler ; methods ; Female ; Glioblastoma ; blood supply ; complications ; diagnostic imaging ; Humans ; Intraoperative Period ; Male ; Middle Aged ; Neovascularization, Pathologic ; diagnostic imaging

OBJECTIVEThe history of clinical application of extracorporeal membrane oxygenation (ECMO) has been more than 30 years. But in China, there were only a few ECMO centers with limited successful cases reported by the end of twentieth century. The high morbidities and mortalities in current pediatric ECMO practice are noted in China. Therefore, it is necessary to review the experience on rescue use of ECMO in critically ill pediatric patients.

METHODA retrospective analysis was done for patients who had been receiving ECMO treatment to rescue refractory cardiorespiratory failure from different causes in a hospital between July 2007 and May 2011.

RESULTA total of 12 patients were treated with ECMO; 7 of them were male and 5 female, they aged 6 days to 11 years, weighed 2.8 - 35 (17.21 ± 11.64) kg. The underlying causes of cardiorespiratory failure were as follows: two cases with acute respiratory distress syndrome (ARDS) leading to respiratory failure, 4 with failure of weaning from cardiopulmonary bypass, 3 with fulminant myocarditis, 1 with right ventricular cardiomyopathy leading to repeated cardiac arrest, 1 with preoperative severe hypoxemia, and 1 with anaphylactic shock complicated with massive pulmonary hemorrhage and severe hypoxemia. Of the 12 cases, 3 were established ECMO (E-CPR) while underwent chest compression cardiopulmonary resuscitation (CPR). The mean ECMO support time was 151.75 (15 - 572) h. Seven patients (58.33%) were weaned from ECMO, 6 patients (50.00%) were successfully discharged. Six cases had bleeding from sutures, 2 cases with severe bleeding underwent thoracotomy hemostasis, 2 presented with acute renal failure. Infection was documented in 3 cases, hyperbilirubinemia in 2 cases, lower limb ischemia in 1 case, hyperglycemia in 3 cases, disseminated intravascular coagulation in 1 case, membrane lung leakage in 2 cases, systemic hemolysis in 3 cases, oxygenator failure in 2 cases and oxygenator thrombosis in one case. During the follow-up between 6 months and 4.5 years, 5 patients survived with good quality of life, without any documented central nervous system disorders. One case survived with the right lower extremity disorder from ischemic damage. His motor function has been improved following orthopedic operation at one year after discharge.

CONCLUSIONECMO is a justifiable alternative treatment for reversible severe cardiopulmonary failure in critically ill children.

Cardiac Output, Low ; etiology ; therapy ; Cause of Death ; Child ; Child, Preschool ; Critical Illness ; mortality ; therapy ; Extracorporeal Membrane Oxygenation ; adverse effects ; Female ; Heart Failure ; etiology ; mortality ; therapy ; Hemorrhage ; epidemiology ; etiology ; Humans ; Infant ; Infant, Newborn ; Male ; Postoperative Complications ; mortality ; therapy ; Respiratory Insufficiency ; etiology ; mortality ; therapy ; Retrospective Studies ; Survival Analysis ; Thrombosis ; epidemiology ; etiology ; Treatment Outcome

BACKGROUNDThe diagnostic value of virtual imaging combined with three-dimensional computed tomographic angiography (3D-CTA) for intracranial aneurysms has not been fully elucidated yet. This study aimed to evaluate the value of combined application of virtual imaging techniques and 3D-CTA in diagnosing patients with aneurismal subarachnoid hemorrhage (SAH) at the acute stage.

METHODSEighty patients with non-traumatic SAH received 3D-CTA examinations. The raw CT data of these patients were reconstructed and transferred into the 3D mode through the surgical plan system based on virtual reality (VR) image, and the 3D virtual images of skulls and brain blood vessels were acquired. The location, size and shape of aneurysms and their anatomic relationship with adjacent tissues were measured from many points of view.

RESULTSSeventy-three aneurysms were detected in 68 of the 80 patients, but 2 aneurysms were detected in 2 of the 5 patients who had been found free of aneurysms previously and had received 3D-CTA examinations for a second time one month later. The 3D virtual images produced by the virtual imaging system were clear and vivid, and they could reveal the location and size of the aneurysm and its relations to the parent artery and skull directly.

CONCLUSIONSThe imaging of 3D-CTA is convenient, reliable and fast in diagnosing intracranial aneurysms and can be regarded as the first choice for the diagnosis and treatment of ruptured intracranial aneurysms. Combined with the surgical plan system based on the VR image, 3D-CTA may obtain more imaging information about aneurysms.

Adolescent ; Adult ; Aged ; Angiography ; methods ; Female ; Humans ; Imaging, Three-Dimensional ; methods ; Intracranial Aneurysm ; diagnostic imaging ; surgery ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods ; Young Adult

BACKGROUNDEarly diagnosis and timely treatment are important for improving therapeutic efficiency of prostate cancer. DNA array is a new bio-technology for disease diagnosis. This study was conducted to diagnose prostate cancer with cDNA macroarray and analysis gene expression profiles of some selective genes in prostate cancer.

METHODSTotal RNA was isolated from patients with prostate cancer and from normal people, and poly (A) RNA was further purified. Then it was analyzed for differentially expressed genes in prostate cancer and normal prostate by cDNA macroarray system.

RESULTSThere were different expressions in the nine prostate-associated specific genes in prostate cancer as compared with normal prostate, in which, 7 were significantly upregulated and 2 were down-regulated.

CONCLUSIONAs a diagnostic approach at molecular level, the cDNA macroarray is an effectively diagnostic method for prostate cancer.

Gene Expression Profiling ; Genes, Tumor Suppressor ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; Prostate-Specific Antigen ; blood ; Prostatic Neoplasms ; diagnosis ; genetics

The study was aimed to investigate the ultranstructural feature and diagnostic criteria of congenital dyserythropoietic anemia-type I (CDA-type I). Nucleated red cells in bone marrow from two patients with CDA-type I were analyzed by transmission electron microscopy (TEM). The results indicated that the erythropoietic/granulopoietic ratio was markedly increased with megaloblastic morphology in all stage of erythrocyte. Most proerythroblast showed of irregular nuclei, while the Swiss-cheese-appearance of the heterochromatin was usually found in basophilic and polychromatic erythroblast. About half of orthochromatic erythroblast illustrated karyolysis and karyorrhexis. Some orthochromatic erythroblast exhibited karyolysis and plasmolysis simultaneously. The inter-nuclear chromatin bridge between separated erythroblasts was seldom found by TEM. The nuclear membrane and rough endoplasmic reticulum were destructed at all stage of erythrocytes in different degree. In conclusion, the megaloblastic erythrosis was the main characteristic of CDA-type I, and then nuclear membrane disruption in polychromatic erythroblast and karyolysis or karyorrhexis in orthochromatic erythroblast. The universal breakdown of cytoplasm membranous system was fundamental pathogenesis of CDA-type I.
Anemia, Dyserythropoietic, Congenital ; blood ; pathology ; Bone Marrow Examination ; Erythroblasts ; ultrastructure ; Erythrocytes ; ultrastructure ; Female ; Humans ; Infant ; Iron ; blood ; Male ; Microscopy, Electron, Transmission

OBJECTIVEThe Ala499Val (C > T) and Lys939Gln (A > C) of the XPC gene are two potentially functional nonsynonymous polymorphisms, which affect the rate of DNA repair and might change XPC production and activity. This study aimed to explore the distribution of these two polymorphisms in the Chinese Han population and their relationship with male infertility.

METHODSWe genotyped the two polymorphisms of the XPC gene by the PCR-restriction fragment length polymorphism (PCR-RFLP) method in 318 infertile patients and 228 fertile male controls, detected the frequency of the alleles, and analyzed both the individual and the joint contribution of the two polymorphisms to male infertility.

RESULTSFor the Ala499Val (C > T) polymorphism, the frequencies of the CC, CT, and TT genotypes were significantly different in distribution between the patients and the controls (P = 0.020). Males with the TT genotype had a lower risk of male infertility than those with the CC genotype (adjusted OR = 0.49, 95% CI: 0.23-0.88), and even lower than those with both CC and CT genotypes (adjusted OR = 0.39, 95% CI: 0.22-0.71). The Lys939Gln (A > C) polymorphism was not related with male infertility. The combined genotype analysis showed that the individuals with 1-4 risk alleles had a significantly higher risk of male infertility (adjusted OR = 2.75, 95% CI = 1.50-5.04) than those with 0 risk allele.

CONCLUSIONThe Ala499Val (C > T) polymorphism of the XPC gene is correlated with male infertility and may be a potential genetic risk factor for male infertility in the Chinese Han population.

Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; DNA Repair ; DNA-Binding Proteins ; genetics ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Infertility, Male ; genetics ; Male ; Polymorphism, Genetic ; Risk Factors

Objective:To perform unilateral patellar resurfacing and contralateral patellar retention in bilateral total knee arthroplasty (TKA) randomly,and to compare the clinical effects of patellar retention with patellar resurfacing in TKA.Methods:In the study,14 bilateral knee osteoarthritis (OA) patients were randomized in the bilateral TKA to receive unilateral patellar resurfacing and contralateral patellar retention,including 28 knees,all were females,53 to 78 years old,with average (66.9 ± 7.8) years,and the BMI was (26.3 ± 1.8) kg/m2.All subjects were followed up from 3 to 12 months.The clinical effects were evaluated based on measurements of American Knee Society score (KSS),range of motion (ROM),anterior knee pain,patellar clunk,and patellar tilt angle (PTA).Results:All the wounds healed primarily without significant complications,such as infection,aseptic loosening,patellar fracture and so on.The preoperative KSS scores of patellar resurfacing group were 38.9 ± 22.2,and the scores changed to be 92.4 ± 6.7 after operation,which were added by 53.5 ± 20.3.While in the patellar retention group,the KSS scores were 38.4 ± 20.5 preoperatively,and after operation,which were added to be 92.1 ±4.2,and improved by 53.7 ±21.4.The differences in the changed KSS scores between TKA with and without patellar resurfacing were not statistically significant (Independent t-test,P =0.98).The ROM was changed from 95.4° ± 13.5° preoperatively to 120.4° ± 8.9° postoperatively in the patellar resurfacing group and from 92.9° ± 19.1 ° preoperatively to 120.4 ± 8.4° postoperatively in the patellar retention group.The ROM of the two group were increased by 25.0° ± 14.5° and 27.5° ± 19.4° re spectively.However,no remarkable differences were observed between the 2 groups in the knee ROM (Independent t-test,P =0.70).At the end of the latest follow-up,3 knees in the patellar resurfacing group and 2 knees in the patellar retention group had knee anterior pain,the incidences of anterior knee pain were 21.4％ and 14.3％ respectively.There was no obvious difference for the incidence of post operative anterior knee pain (Chi-square test,P =0.62).The incidences of post-operative patellar clunk in the 2 groups were all with 3 knees (21.4％),which had no significant difference in the 2 groups (Chi-square test,P =1.00).The post-operative PTA were 2.6° ± 2.6° in the patellar resurfacing group and 3.6° ± 2.9° in the patellar retention group,respectively.There was also no statistical difference between the 2 groups (Chi-square test,P =0.36).Conclusion:For knee OA patients with mild or moderate patellar cartilage damage,performing patellar resurfacing or not didn't significantly affect anterior knee pain,patellar clunk,functional outcomes or patellar tracking after TKA.So we suggest retain patella in TKA for OA patients with mild or moderate patellar cartilage damage.

Objective To evaluate the occurrence and management of complications following mini- mally invasive percutaneous nephrostolithotomy (MPCNL).Methods The data of 436 cases of MPCNL from December 2001 to March 2005 were reviewed,including 249 male cases and 187 female cases.Their age ranged from 14 to 71 years with a mean of 41.6 years.Of the 436 cases,314 cases had simple nephrolith- iasis,79 had nephrolithiasis combined with upper ureterolithiasis,27 had unilateral upper ureterolithiasis,and 16 had bilateral upper ureterolithiasis.Results Among the 436 cases of MPCNL,complications occurred in 27 cases (6.2%).Of the 27 cases,5 had massive hemorrhage,which was cured by closing meatus of nephric fistula,cleaning out the blood clot within the bladder with instrument and transfusing blood;2 cases had pleural effusion,11 cases had hydroperitoneum,8 eases had renal perforating injury,and they were cured by puncture and drainage;1 case had pyocalix,which was cured by using effective antibiotics,re-punc- turing kidney and postponing pulling out the fistula catheter.Conclusions MPCNL is a minimally inva- sire operation.However,it has obvious risk if the surgeon's skill is not proficient.The improvement in the pre- vention and management of complications can promote the application of this procedure.

BACKGROUNDWith conventional imaging methods only the morphous of the visual nerve fiber bundles can be demonstrated, while the earlier period functional changes can not be demonstrated. We hypothesized that diffusion tensor imaging (DTI) would demonstrated the whole optic never fiber bundle and visual pathway and the earlier period functional changes. The purpose of the present study was to evaluate the application of DTI technique in the demonstration of the whole optic never fiber bundle and visual pathway, and the influence of orbital tumors on them.

METHODSGE 1.5 T signa HD MR System, and the software package DTV2 were adopted. The total 45 subjects were enrolled, including 15 volunteers and 30 patients. All patients had ocular proptosis from minor to major. Seven patients had visual acuity decrescence.

RESULTSThe nerve fiber bundles, e.g. optic chiasma, optic tract and optic radiation in posterior visual pathway were well demonstrated in all cases. Wherein, the intact whole visual pathway fiber bundles were clearly revealed in 10 volunteers and 17 patients, and optic nerve was not wholly revealed in the rest of the subjects. Shift of optic nerve caused by compression and partial deformation were seen in 7 patients with orbital tumor. In 6 of 7 patients, DTI displayed significant abscise and deformation of visual nerve. Chi-square test indicated significant correlation between visual acuity decrescence and DTI visual nerve non-display.

CONCLUSIONSVisual nerve fiber bundles and the whole visual pathway were visualized in most of patients with DTI. It might be an effective method of providing imaging evidence for visual nerve fiber earlier period functional changes, and laid a foundation for the study in other cranial nerves.

Adult ; Aged ; Diffusion Magnetic Resonance Imaging ; methods ; Exophthalmos ; pathology ; Female ; Humans ; Male ; Middle Aged ; Optic Chiasm ; anatomy & histology ; pathology ; Optic Nerve ; anatomy & histology ; pathology ; Visual Pathways ; anatomy & histology ; pathology ; Young Adult