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.

Objective To study the macrophage colony-stimulating factor(M-CSF)expression levels of serum and synovial fluids from patients with spondyloarthropathy(SPA)and its contribution to the pathogen- esis of SpA.Methods Eleven SpA synovial tissue samples were compared to those from peripheral blood mononuclear cells(PBMC)of 10 normal subjects using a 1176 gene array.M-CSF was detected in both serum samples and synovial fluids by enzyme linked immunosorbent assay(ELISA).Two groups of AS subjects were tested.The first group consisted of 41 ankylosing spondylitis(AS)patients who had not been treated with bio- logics.The second group consisted of 13 subjects whose serum samples were collected before and 14 weeks af- ter initiation of infliximab.These were compared to serum samples from 28 normal subjects,and synovial fluid samples from 15 SpA patients.Results Expression of M-CSF could be detected in both serum samples and synovial fluids.The concentration of M-CSF in the group of 41 AS patients not treated with biologics correlated with the Bath Ankylosing Spondylitis Disease Activity Index(BASDAI)values(r=0.41,P=0.004).Treatment of infliximab in AS patients led to a significant decrease in the values of BASDAI(P=0.000 07),but no signif- icant change in the serum M-CSF values.Conclusion M-CSF is a promising candidate for research on the mechanisms of SpA and its signaling on pathway in SpA is different from tumor necrosis factor(TNF)-?,and it may provide new basis for developing new anti-biologics for SpA.

OBJECTIVETo analyze the magnetic resonance imaging (MRI) characteristics of neuromyelitis optica (NMO) in Chinese patients.

METHODSWe retrospectively reviewed the MRI films of 61 patients with NMO (including 57 female and 4 male patients) admitted in our department.

RESULTSOf these patients, 39 (79.6%) showed positivity for serum aquaporin-4 (AQP4) antibody. On MRI, 18 patients showed involvement of the cervical cord alone, 27 had both cervical and thoracic segment involvement, and 16 displayed thoracic segment involvement. The lesions appeared linear (9 cases), diffuse (23 cases), or both (29 cases), mostly located axially with occasional lateral distribution. Thirty-nine of the 61 patients (63.9%) had brain abnormalities, 31 presented with supratentorial lesions (mostly in the juxtacortical, subcortical, deep white matter and lateral ventricle-adjacent regions, n=27), 15 showed infratentorial lesions (mostly in the preiaqueduct-fourth ventricular-central canal, n=13), and 7 had supra- and infratentorial lesions simultaneously.

CONCLUSIONNMO has complex MRI presentation, and linear lesions in the spinal cord and preiaqueduct-fourth ventricular-central canal lesions, where AQP4 is high expressed, can be characteristic for NMO. MRI and AQP4 antibody detection are suggested for suspected cases for early diagnosis.

Adult ; Aquaporin 4 ; immunology ; Brain ; pathology ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Neuromyelitis Optica ; pathology ; Retrospective Studies ; Spinal Cord ; pathology

OBJECTIVETo assess the seroprevalence and diagnostic value of aquaporin-4 antibody (AQP4-Ab) in patients with inflammatory central nervous system demyelinating diseases.

METHODSSeventy-two patients with neuromyelitis optica (NMO), 68 with multiple sclerosis (MS), 4 with optic neuritis (ON), and 41 with transverse myelitis (TM) were included in this study. The TM group comprised 19 patients with non-longitudinally extensive transverse myelitis (nLETM), 14 with monophasic longitudinally extensive transverse myelitis (mLETM), and 8 with recurrent longitudinally extensive transverse myelitis (rLETM). The serum levels of AQP4-Ab was detected by indirect immunofluorence assay in these patients.

RESULTSAQP4-Ab was detected in 72.2% (52/72) patients with NMO, 5.9% (4/68) patients with MS, 25.0% (1/4) patients with ON, and 17.1% (7/41) patients with TM, showing a significant difference in the positivity between NMO and MS groups (P<0.01). AQP4-Ab seropositivity rate was 5.3% (1/19) in nLETM patients, 62.5% (5/8) in rLETM patients and 7.1% (1/14) in mLETM patients, significantly higher in rLETM than in nLETM (P<0.01) and mLETM groups (P<0.05), but no statistical difference was found between rLETM and NMO groups.

CONCLUSIONSA high seroprevalence of AQP4-Ab is observed in patients with NMO and rLETM, which support the hypothesis that NMO and rLETM belong to NMO spectrum disorders. AQP4-Ab can serve as a useful index for diagnosing NMO and differential diagnosis from MS. More attention and effective immunosuppressive treatments should be given to patients positive for AQP4-Ab.

Aquaporin 4 ; immunology ; Autoantibodies ; blood ; Demyelinating Autoimmune Diseases, CNS ; diagnosis ; immunology ; Female ; Humans ; Male ; Multiple Sclerosis ; diagnosis ; immunology ; Neuromyelitis Optica ; diagnosis ; immunology ; Seroepidemiologic Studies

Objective To explore the effects of radiofrequency catheter ablation(RFCA)on the blood coaguable states and the clinical value of perioperative plasma D-dimer.Methods The plasma level of D-dimer was assayed by enzyme-linked immunosorbent assay(ELISA)in blood samples of 30 children who were undertaken RFCA.Blood samples were consecutively obtained before cannulating,after electrophysiologic(EP)study,immediately after RFCA,the second day and the seventh day after RFCA.The centrifuged spead was 3 000 r/min,keep it for 10 minutes to obtain the upper plasma,and the crvopreserve.Results The plasma levels of D-dimer was highest at the time point when RFCA was successfully accomplished and restored to preoperative level in the seventh day after RFCA.There were statistically significant difference in the paried values at different time points(Pa

Objective:To establish a human lung squamous carcinoma cell line and to study its biological characteristics. Methods:Lung squamous carcinoma specimens were freshly resected during operation;the tissues were incubated in vitro and the cell line was named CHLH-1.The biological characteristics of the cells were studied by light microscopy,electron microsco- py,chromosome analysis and transplantation experiment.Results:Cells from the specimens of the primary tumor,the CHLC- 1 cell line and the cells from transplanted tumor possessed the characteristics of malignant squamous epithelium under light and electron microscope.The cell growth curve,doubling time and mitotic index were also observed in vitro.Nuclear chromosome analysis revealed that the tumor was a subtriploid with a mode of 60-68 per cell.Tumor nodes were observed under the skin of nude mice by heterogenic transplantation.Conclusion:The characteristics of the established cell line suggest that it is a newly established human squamous carcinoma cell line.

OBJECTIVETo investigate the effects of microRNA-144 (miR-144) expression on H9C2 (2-1) myocytes.

METHODSMiR-144 was up-regulated in primary cultured H9C2 (2-1) myocytes through transfection. Cells transfected with Lipofectamine(TM) 2000 and its mixture with miRNA synthesized randomly as blank control and negative control respectively. The up-regulation of miR-144 was confirmed by real-time PCR. Cell apoptosis was evaluated by means of CCK-8, Caspase-3 and flow cytometry.

RESULTSReal-time PCR results showed that the miR-144 expression was obviously increased in miR-144 up-regulation group (2178.84 ± 838.52) compared with negative (2.06 ± 0.73) and blank (1.00 ± 0.00) control group (all P < 0.01). The proliferation was lower, the activity of Caspase-3 was elevated and the apoptosis rates were significantly increased in miR-144 up-regulation group compared with negative and blank control group, while no significant difference was found between the latter 2 groups.

CONCLUSIONMiR-144 mimics may selectively up-regulate the expression of miR-144 in myocardial cells and consequently promote apoptosis and inhibit proliferation in myocardial cells.

Animals ; Apoptosis ; genetics ; Caspase 3 ; metabolism ; Cell Line ; MicroRNAs ; genetics ; metabolism ; Muscle Cells ; metabolism ; Rats ; Sincalide ; metabolism ; Transfection

Objective To study the chest image appearances of penicilliosis marneffei(PSM)in patients with acquired immune deficiency syndrome(AIDS).Methods Chest imaging features of PSM in 36 patients with AIDS were retrospectively analyzed.Results Radiographic features of infiltrative lesions and focal lung consolidation were found in 14 cases(38.89%),in which 2 cases were with single lung disease(5.56%)and 12 cases with bilateral lung involvement(33.33%).Eight cases had diffuse lesions (22.22%),10 cases had reticular image patterns(27.78%),9 cases had nodular patterns(25.00%), 7 cases had ground-glass shadows(19.44%),6 cases had diffuse miliary lesions(16.67%),4 cases had enlarged bilar and enlarged mediastinum lymph nodes(11.11%).Cystic lesions was found in 5 cases (13.89%).Four cases had pleural effusion(11.11%),and 2 cases had nodular bump(5.56%). Pericardial effusion and pneumothorax each appeared in 1 case(2.78%).By HRCT,infiltrative lesion and focal lung consolidation were found in 32 patients(88.89%),in which 4 cases were with single lung lesions (11.11%)and 28 cases were with bilateral lung lesions(77.78%).Thirteen cases had diffuse lesions (36.11%),10 cases had pulmonary interstitial hyperplasia(27.78%),9 cases had nodular patterns (25.00%),8 cases had ground-glass shadows(22.22%),9 cases had diffuse miliary lesions(25.00%), 21 cases had enlarged lymph nodes in the mediastinum(58.33%).Cystic lesions were found in 8 cases (22.22%).Thirteen cases had pleural effusion(36.11%),and 2 cases had nodular bump(5.56%). Pericardial effusion and pneumothorax each appeared in 1 case(2.78%).Conclusion The image appearances of PSM in patients with AIDS include infiltrative lesions or focal lung consolidation,ground- glass shadow,enlarged hilar and mediastinum lymph nodes,pleural effusion,interstitial involvement or reticular image pattern(pulmonary interstitial hyperplasia),diffuse miliary lesion,and cystic lesion.

OBJECTIVETo investigate the effect of Rongshi granule on osteopontin(OPN) expression.

METHODThe urlisthiasis rats were induced by ethylene glycol (EG) and ammonium chloride, the control group rats were non-treated, and the Rongshi granule groups (low-dose group, middle-dose group and high-dose group) were administered Rongshi granule in addition to EG and ammonium chloride in 21 days. Pooled 24 h urine samples from each group were collected weekly with the use of metabolic cages, the concentration of uric calcium and oxalic acid were respectively measured by EDTA and photoelectric colorimetric method. Eight animals from each group were killed at 0, 7, 14, and 21 days, kidneys were histologic examinaed and immunohistochemical staining.

RESULTThe expression of kidney osteopontin in model group was obviously higher than that of control group (P <0.01), and was up to the highest at 21 days with 1.4 times (0.281 3/0.201 8) of the control group. The expression of kidney osteopontin in all of the Rongshi granule groups were lower than those of model group (P < 0.05), with an obvious dose-dependent manner. The degree of the kidney calcium oxalate crystal of the rats in all the Rongshi granule groups was much lower than that of model group, and the uric calcium and oxalic acid were much lower than those of model group (P < 0.01).

CONCLUSIONThe Rongshi granule could inhibit the expression of osteopontin in rat urolithiasis model.

Ammonium Chloride ; Animals ; Calcium ; urine ; Calcium Oxalate ; metabolism ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Ethylene Glycol ; Female ; Kidney ; metabolism ; Kidney Calculi ; chemically induced ; metabolism ; Male ; Osteopontin ; metabolism ; Oxalic Acid ; urine ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the clinical feature of extracranial and intracranial complications caused by cholesteatoma otitis media.

METHODSA retrospective study as made of 78 patients with extracranial and intracranial complications caused by suppurative otitis media between January 1992 and December 2002 was conducted.

RESULTSFifty-six of the 78 patients had one complication and the other 22 patients had two or more complications among them. In this series, 24 cases had facial paralysis, 16 cases with suppurative labyrinthitis, 12 cases with meningitis, 11 cases with periauricular abscess, 9 cases with extradural abscess, 9 case with otogenic meningitis brain abscess, 6 case with lesion of sigmoid sinus and 5 cases with Bezold abscess. Of the 24 patients with facial paralysis, 15 with continued nerves had obtained omatomic preservation of the facial nerves, The nerve function of 11 patients recovered from House-Brackmann grade IV to grade II, the rest 4 patients didn't regain after operation. Four of the 9 patients with their facial nerves broken received anastomosis of the facial nerves, three cases received transplantation of the nervus auricularis magnus, and two received anastomosis to nervus auricularis magnus. The nerve function recovered from House-Brackmann grade V - IV to grade III.

CONCLUSIONSEven though the incidence of the complications has been low down in past years, there are still some serious complications. Temporal bone CT scan plays an important role in the diagnosis of extracranial and intracranial complications of cholesteatoma otitis media. The most reliable method to diagnosis and treat the complications is surgery.

Adolescent ; Adult ; Aged ; Brain Diseases ; etiology ; Child ; Child, Preschool ; Cholesteatoma, Middle Ear ; complications ; Facial Paralysis ; etiology ; Female ; Humans ; Intraoperative Complications ; Male ; Middle Aged ; Postoperative Complications ; Retrospective Studies ; Young Adult