Animals ; Bile Duct Neoplasms ; chemistry ; Bile Ducts, Intrahepatic ; Cholangiocarcinoma ; chemistry ; Cricetinae ; Epidermal Growth Factor ; analysis ; Immunohistochemistry ; Receptor, Epidermal Growth Factor ; analysis ; Transforming Growth Factor alpha ; analysis

An expression plasmid carrying anthrax protective antigen (PA) gene was constructed, which has an OmpA signal sequence attached to the 5' end of PA gene. The plasmid was transformed into E. coli and induced to express recombinant PA (rPA) . The recombinant protein, about 10% of the total bacterial protein in volume, was secreted to the periplasmic space of the cell. After a purification procedure including ion-exchange, hydrophobic interaction chromatography, and gel filtration, about 15 mg of 95 % pure rPA was obtained from 1-liter culture. The bioactivity of rPA was proved by in vitro cytotoxicity assay. The polyclonal antiserum from rabbits immunized with rPA could inhibit the action of anthrax lethal toxin in vitro, which suggests that antibodies against rPA can provide high passive protection against anthrax. The results reported here may be helpful to develop a safe and efficacious recombinant PA vaccine against anthrax.
Amino Acid Sequence ; Animals ; Anthrax Vaccines ; immunology ; Antigens, Bacterial ; chemistry ; genetics ; immunology ; toxicity ; Bacterial Toxins ; chemistry ; genetics ; immunology ; toxicity ; Base Sequence ; Mice ; Molecular Sequence Data ; Plasmids ; Rabbits ; Recombinant Proteins ; biosynthesis ; immunology ; Vaccines, Synthetic ; immunology

The study was aimed to investigate the influence of penicillin and streptomycin on proliferation, apoptosis and extracellular secretion (ECS) produced from human umbilical cord derived mesenchymal stem cells (MSC). MSC were isolated from umbilical cord tissue, then the immunotyping, multipotent differentiation and proliferation of these cells were assayed by cytometry, cytochemistry and MTT respectively. The expressions of ECS and apoptosis-related genes (bcl-2, bax) were detected by quantitative RT-PCR. The results showed that the phenotype of these cells matched with the characteristics of MSC. Penicillin and streptomycin of low concentrations promoted MSC proliferation, with the most effective concentration of 100 U/ml. Expressions of ECS cultured in addition of penicillin and streptomycin were down-regulated. Furthermore, apoptosis-related factor (bcl-2/bax) expression levels in low concentrations penicillin and streptomycin groups were higher than that in the control group. It is concluded that low concentrations penicillin and streptomycin can promote the proliferation and reduce the apoptotic rate, but high dose can inhibit the ECS component expression of MSC.
Cell Differentiation ; Cells, Cultured ; Extracellular Matrix ; secretion ; Flow Cytometry ; Gene Expression Regulation ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; secretion ; Penicillins ; pharmacology ; Streptomycin ; pharmacology ; Umbilical Cord ; cytology

BACKGROUNDTo investigate the prevalence of anti-HEV among swine, sheep and chickens.

METHODSTotally 498 sera of swine, sheep and chickens collected from Xingjiang, Guangxi, Guangdong, Beijing and Hebei were detected for the anti-HEV by an enzyme linked immunoassay.

RESULTSThe anti-HEV positive rate of swine was 67.53%(104/154), in pigs between 4-5 months of age the rate was 100.00%(9/9) from Xingjiang. The rate in pigs under 3 months of age from Guangxi was 36.00%(9/25) and in pigs older than six months of age was 71.67% (86/120), respectively. The 108 sera of sheep collected from Xingjiang were all negative. The positive rate of chickens was only 1.27% (3/236). The anti-HEV prevalence rates of chickens from Luoding, Shenzhen, Liuzhou, Beijing and Hebei were 4.00%, 1.49%, 1.49%, 0, 0 respectively.

CONCLUSIONHEV infection does exist among swine and chickens. The anti-HEV prevalence of swine was the highest among domestic animals. The role of swine and chickens in transmission of HEV needs to be further studied.

Animals ; Antibodies, Viral ; Chickens ; China ; epidemiology ; Hepatitis Antibodies ; blood ; Hepatitis E ; epidemiology ; veterinary ; Hepatitis E virus ; immunology ; Poultry Diseases ; epidemiology ; virology ; Prevalence ; Sheep ; Sheep Diseases ; epidemiology ; virology ; Swine ; Swine Diseases ; epidemiology ; virology

OBJECTIVESTo study the effect of testicular local heating on spermatogenic cell apoptosis in rat.

METHODSSeventy male SD rats were divided into heat treatment group (43 degrees C) and control group (22 degrees C). Each group was further divided into seven sub-groups respectively according to the time of 12 hours and 1 days, 3 days, 6 days, 10 days, 50 days and 80 days after testicular local treatment. The spermatogenic cell apoptosis in all sub-groups was examined by means of electron microscopy, flow cytometry and terminal deoxynucleotidyl transferase-mediated dUDP-nick end labeling(TUNEL) method.

RESULTSIn the groups of heat treatment, spermatogenic cell apoptosis was detected by electron microscopy; flow cytometry showed that the percentage of cells with sub-haploid increased(P < 0.01); the percentage of positive TUNEL cells in the heat treatment groups was higher than that in the control group(P < 0.01). Initiation of spermatogenic cell apoptosis after testicular heating was not random but was highly selective.

CONCLUSIONSLocal testicular heating could increase the spermatogenic cell apoptosis. The most sensitive cell is spermatocyte. Spermatid and sperm also display apparent changes. Heating can increase the apoptosis of spermatogonia in a long period.

Animals ; Apoptosis ; Flow Cytometry ; Hot Temperature ; In Situ Nick-End Labeling ; Male ; Microscopy, Electron ; Rats ; Rats, Sprague-Dawley ; Spermatogonia ; cytology ; ultrastructure ; Testis ; pathology ; ultrastructure

OBJECTIVETo evaluate the applicability of the polymorphic marker closely linked with beta-globin gene for the preimplantation genetic diagnosis (PGD) in couples at risk of having child with beta-thalassemia.

METHODSSingle cell multiplex nested PCR which coamplifies the beta-globin gene and the closely linked polymorphic marker, HumTHO1 gene, was applied in six clinical PGD cycles for four couples with beta-thalassemia.

RESULTSIn six clinical PGD cycles, a total of 44 embryos were biopsied and 44 blastomeres were obtained. Forty-one blastomeres were amplified and thirty-five embryos were given definite diagnoses. Fourteen embryos were transferred back to the uterus of the patients and one pregnancy went on well and ended with one live healthy birth, which confirmed the results of PGD. The average amplification efficiency of single blastomere was 89.7% and the average allele drop-out(ADO) rate was 14.4%. The coamplification of HumTHO1 could help to detect the existence of ADO and contamination.

CONCLUSIONThis is the first report on unaffected pregnancy resulting from PGD using multiplex nested PCR in China. The simultaneous amplification of polymorphic marker closely linked to beta-globin gene(HumTHO1) could help to resist the risk of misdiagnosis in PGD caused by ADO and contamination.

Adult ; Female ; Humans ; Male ; Polymerase Chain Reaction ; Pregnancy ; Preimplantation Diagnosis ; methods ; beta-Globins ; genetics ; beta-Thalassemia ; diagnosis ; genetics

OBJECTIVETo investigate the application of penile cavernous bodies elongation combined with fat flap for the treatment of micro-penis.

METHODSAnatomic study was performed to study the thickness of penile suspension ligaments and the relationship between the penile erection stability and the mobilization of cavernous bodies crus. The suspension ligaments were divided and cavernous bodies crus were partially mobilized, so as to release part of the cavernous bodies from inferior ramus of pubis. Then the penis was elongated sufficiently. Local fat flap was transposed to fill the front space of pubis to make sure the effective elongation of penis.

RESULTS205 cases of micro-penis were treated. The average length of the penis was 4.26 cm in the static state, 8.13 cm in erectile state before operation. After operation, it increased to 8.63 cm in the static state, 12.11 cm in erectile state.

CONCLUSIONSThe cavernous bodies can be elongated 1-2 cm more with the modified method, while the stability of penile erection is not affected.

Adolescent ; Adult ; Aged ; Humans ; Male ; Middle Aged ; Penis ; anatomy & histology ; surgery ; Surgical Flaps ; Treatment Outcome ; Young Adult

BACKGROUNDAlthough endomyocardial biopsy (EMB) plays a crucial role in the final diagnosis in patients with heart failure of unknown etiology, the invasive nature of this technique limits its clinical application in China. The purpose of this study was to evaluate the clinical application of EMB in diagnosing cardiomyopathy with unexplained etiologies in China.

METHODSFifty-three consecutive patients (38 males, age 14 - 67 years, median 43 years) were included in the study who were initially diagnosed as unexplained cardiomyopathy and under EMB biopsy in Peking Union Medical College Hospital from 2006 to 2009. The patients were clinically divided into four groups: dilated, hypertrophic, restrictive and unclassified cardiomyopathy. Biopsies were performed via right internal jugular vein with the use of the bioptome under fluoroscopic guidance. Three to five endomyocardial samples were taken from each patient for light microscopy examination and one sample for electron microscopy was taken if necessary. For each patient, an initial clinical diagnosis, an EMB diagnosis and a final diagnosis prior to discharge were established. All the data were compared and analyzed for the evaluation of clinical utility of EMB in China.

RESULTSIn 26 patients initially diagnosed with restrictive cardiomyopathy (RCM), the etiology of the condition was finally diagnosed using EMB in 15; including 13 amyloidosis and two eosinophilic myocarditis. We employed EMB in 19 patients clinically diagnosed as dilated cardiomyopathy and detected viral myocarditis in one patient, cardiac involvement due to polymyositis in four and doxorubicin-induced cardiomyopathy in one. In five patients with severe left ventricle hypertrophy undergoing EMB, one patient was diagnosed as autophagic vacuolar cardiomyopathy and one as mitochondrial disease. In the remaining three patients with unclassified cardiomyopathy, EMB revealed infiltration of eosinophils as the cause of atrial ventricular block in one patient. Final diagnoses were made in 24 of the total 53 patients (45%) based on the combination of EMB and clinical data. Transient atrial ventricular block in a patient with prior complete left bundle branch block was the only complication occurred during the procedures.

CONCLUSIONThe clinical application of EMB is safe. The combination of EMB and clinical data produced a better understanding of the mechanisms behind the clinically diagnosed cardiomyopathy in China.

Adolescent ; Adult ; Aged ; Biopsy ; methods ; Cardiomyopathies ; classification ; diagnosis ; pathology ; Cardiomyopathy, Dilated ; diagnosis ; pathology ; Cardiomyopathy, Hypertrophic ; diagnosis ; pathology ; Cardiomyopathy, Restrictive ; diagnosis ; pathology ; Female ; Humans ; Male ; Middle Aged ; Myocardium ; pathology ; Young Adult

OBJECTIVETo observe the clinical features and cardiac magnetic resonance imaging (CMR) characteristics of patients with endomyocardial biopsy (EMB)-proven cardiac amyloidosis (CA).

METHODSEMB proven CA patients underwent CMR examination from September 2006 to December 2010 were included. The findings of clinical manifestation, electrocardiogram, echocardiography and CMR were analyzed.

RESULTSAmong the 18 patients with EMB verified CA, 5 patients underwent CMR. All 5 patients had heart failure symptoms and electrocardiogram was abnormal. Echocardiogram showed concentric left ventricular hypertrophy, granular appearance of the myocardium, left atrial enlargement and moderate to severe left ventricular diastolic dysfunction. CMR revealed increased thickness of the left ventricular wall (especially at the inter-ventricular septum), enlarged bilateral auricle, restricted left ventricular filling with normal or mild to moderate reduced systolic function. Pleural and pericardial effusions were observed in 2 patients. Abnormal late gadolinium enhancement (LGE) was detected in all 5 patients. CMR revealed different patterns of LGE. Left ventricular global subendocardial delayed gadolinium enhancement or transmural delayed gadolinium enhancement were found, and patients also showed line-, granular- or patchy-like enhancement. The degree and range of LGE paralleled the disease course and were consistent with electrocardiogram changes.

CONCLUSIONSAs a noninvasive diagnostic tool, CMR is valuable in the diagnosis of CA. For patients with clinical suspicion of CA, CMR could be a helpful diagnostic tool, especially in the hospitals where EMB is not available.

Amyloidosis ; diagnosis ; Biopsy ; Cardiomyopathies ; diagnosis ; Echocardiography ; Electrocardiography ; Gadolinium ; Gadolinium DTPA ; Humans ; Hypertrophy, Left Ventricular ; Magnetic Resonance Imaging ; Myocardium ; Systole

OBJECTIVETo investigate the prevalence of peripheral arterial disease (PAD) in China type 2 diabetic patients and to demonstrate the relationships between putative risk factors and PAD.

METHODSIn total 1,397 type 2 diabetic patients aged 50 years and older were enrolled and determined ankle-brachial index (ABI) and brachial-ankle pulse wave velocity (baPWV) in 15 Class III Grade A hospitals in 7 major cities of China.

RESULTSMean patient age was 63.7 +/- 8.2 years and mean duration of diabetes mellitus was 9.39 +/- 7.4 years. Two hundreds and seventy-two (19.47%) patients were diagnosed as PAD by ABI < 0.9, 122 (18.37%) in male and 150 (20.46%) in female. PAD patients had a significantly longer duration of diabetes mellitus, higher hemoglobin A1c, and a significantly lower mean body mass index than non-PAD ones. Aging, smoking, and systolic blood pressure were found to be positively related with the prevalence of PAD. In terms of lipid profiles, no variable was found to relate with PAD. Notably, baPWV showed as the same significant guiding index for PAD, almost matched with ABI.

CONCLUSIONSPAD is a common complication in China type 2 diabetic patients. Therefore, PAD screening and treatment should be emphasized for diabetic patients with high risk factors.

Aged ; China ; epidemiology ; Diabetic Angiopathies ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Peripheral Vascular Diseases ; epidemiology ; Prevalence ; Risk Factors ; Urban Population ; statistics & numerical data