Purpose To indicate the expression of nattokinase (NK) in Pichia pastoris , an emulsion was prepared with the purified NK to prepare polyclonal antibody. In order to establish sandwich enzyme-linked immunosorbent assay (ELISA) to assay NK in organism, furthermore to lay the foundation for researching in vivo metabolism and function of NK. Methods The NK gene was cloned into a Pichia pastoris expression vector pHBM905A to construct the recombinant plasmid pPRONK.The recipient cell of Pichia pastoris GS115 was transformed with pPRONK which had been cut by restriction enzyme Sal I , under the induction of methanol. The expressed production is purified by salting out and ultrafiltration membrane. An emulsion was prepared with the purified NK and injected into rabbits to prepare polyclonal antibody. Results NK was expressed and identified by SDS-PAGE.The molecular mass of expressed production is about 27 kD.The fibrin plate assay indicated that the NK protein can cleavage fibrin effectively. ELISA analysis indicated that the polyclonal antibody titer is about 1:8 000. Western blot demonstrated that there was a special strap nearby 27 kD. Conclusion NK was successfully expressed in Pichia pastoris , the production can cleavage fibrin effectively and it had great immunogenicity.

Adult ; Aged ; Breast Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Female ; Gene Amplification ; Genital Neoplasms, Male ; genetics ; metabolism ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Paget Disease, Extramammary ; genetics ; metabolism ; pathology ; surgery ; Paget's Disease, Mammary ; genetics ; metabolism ; pathology ; surgery ; Penile Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Receptor, ErbB-2 ; genetics ; metabolism ; Scrotum ; Vulvar Neoplasms ; genetics ; metabolism ; pathology ; surgery

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Paraquat ; poisoning

OBJECTIVE:To investigate the effects of selenium(Se)and/or vitamin E(VE)on the NO and NOS in heart,liver,kidney and serum of experimental hyperlipidemic rats.METHODS:SD rats were divided into5groups,administreated by Se and/or VE.After4weeks,the NO contents and NOS activities in heart,liver,kidney and serum were assayed by NO kit and NOS kit respectively.RESULTS:NO contents and NOS activities could be reduced in heart,liver,but increased in serum and kidney by high-fat feed(HFF).Meanwhile,VE and/or Se could increase the NO contents in all the experimental samples and NOS activities in heart,liver and kidney(P

Objective To investigate the relationship between epigenetic changes involving multiple modifications of histones and both happening of glioma and pathological grades of glioma.Methods Immunohistochemistry was performed on 67 samples that were cut off from patients with gliomas who made their definite diagnosis from 2006 to 2008 to evaluate the level of histone 4 lysine 12 acetylation (H4K12Ac), histone 4 arginine 3 monomethylation (H4R3monoMe) and histone 4 lysine 20 acetylation (H3K18Ac). Results H4K12Ac, H4R3monoMe and H4K20triMe were all detected at high frequencies in tumor tissue of glioma. The expressions of H4K12Ac and H4R3monoMe were positively correlated with increasing WHO pathological grades (P＜0.05), while H4K20triMe staining was not significantly correlated with WHO pathological grades (P＞0.05). Conclusion Multiple modifications of histones may affect several steps in tumor cell biology, and may have a prognostic value in gliomas since specific modifications were correlated with WHO pathological grades of glioma.

Adult ; Diagnosis, Differential ; Female ; Humans ; Hysterectomy ; Leiomyomatosis ; pathology ; surgery ; Sarcoma, Endometrial Stromal ; pathology ; Uterine Neoplasms ; pathology ; surgery ; Uterus ; blood supply ; Vascular Neoplasms ; pathology ; surgery

To evaluate the regulating effect of Buyang Huanwu decoction and its simple prescription (Naojian tablet) on CDK4/Cyclin D1 expression in hippocampus tissues of rats with cerebral ischemia, SD rats were divided into the sham-operation group, the model group, the Buyang Huanwu decoction group (ig, 3.15 g · kg⁻¹) and the simple prescription group (ig, 2.41 g · kg⁻¹). Each group was further divided into five subgroups based on time points after the administration, i. e. 1 d, 3 d, 7 d, 14 d and 28 d, respectively. CDK4/Cyclin D1 expressions of the group at different time points were examined by using immunohistochemistry and real-time qPCR. According to the results, the cerebral ischemia model group showed higher CDK4/Cyclin D1 expression than the sham-operation groups (P < 0.05), suggesting that the cell cycle signal pathway would be activated by the cerebral ischemic injury. Both Buyang Huanwu decoction and simple prescription groups showed significantly lower cyclin expression than the model group at 3 d, 7 d, 14 d, 28 d (P < 0.05), indicating both Buyang Huanwu decoction and its simple prescription could play the neuroprotective effect through the cell cycle signal pathway.
Animals ; Brain Ischemia ; drug therapy ; genetics ; metabolism ; physiopathology ; Cell Cycle ; drug effects ; Cyclin D1 ; genetics ; metabolism ; Cyclin-Dependent Kinase 4 ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Rats ; Rats, Sprague-Dawley

Adult ; Burns ; etiology ; Humans ; Male ; Middle Aged ; Paraquat ; adverse effects ; Scrotum ; injuries

OBJECTIVETo observe the clinical therapeutic effect of Nuzhen Yunyu Decoction (NYD, a Chinese recipe for nourishing blood and reinforcing Shen) and its influence on the hemodynamic parameters in ovarian and uterine arteries.

METHODSSixty-eight patients were randomly divided into two groups at the ratio of 2:1, 45 in the TCM group treated with Chinese medicine and 23 in the control group treated with clomiphene citrate. Menstrual cycle were taken as the indexes for clinical observation; basal body temperature, cervical mucus, vaginal exfoliative cells, serum hormones and B ultrasonic examination were taken as the indexes for ovulation and pregnancy assessment. The hemodynamic parameters of uterus and ovary were determined by using color Doppler ultrasonography, and the growth of follicles and ovulation, thickness of uterine endometrium were also monitored continuously.

RESULTSResults of 3 cycles treatment and 1 year follow-up study showed that the ovulation promoting rate, pregnancy rate and abortion rate in the two groups were similar, but the treated group showed better effects than that of control group, in such aspects as regulating menstruation, promoting growth and development of follicle, strengthening endometrium, improving blood supply and circulation of uterine and ovary (P < 0.05, P < 0.01).

CONCLUSIONNYD has the effects of ovulation promotion and holistic regulation of reproductive system.

Adult ; Anovulation ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hemodynamics ; Humans ; Infertility, Female ; drug therapy ; etiology ; Ovary ; blood supply ; Ovulation Induction ; Phytotherapy ; Uterus ; blood supply ; Yin Deficiency ; drug therapy

Objective To investigate the relevant risk factors for fungal infection following operation of the gastrointestinal neo- plasm and offer supporting data for the prevention of fungal infection.Methods Medical records from 116 patients who under- went the operation of gastrointestinal neoplasm in the special group of this hospital from January 2006 to June 2006 were retro- spectively reviewed on the relevant risk factors by univariate and multivariate Logistic regression analysis.Results Of the 116 patients reviewed, 18 had fungal infection.Forty-six samples were positive for fungal pathogen.The most frequently isolated fungal strain was Candida albicans (15/20) and the most common infection site was gastrointestinal tract (14/18).Fungal in- fection after the operation of gastrointestinal neoplasm was significantly relevant with the duration of antibiotic use, duration of post-operative fasting, low serum albumin, high blood glucose and complication of bacterial infection.The duration of antibiotic use was a significantly independent risk factor.Conclusions Reasonable antibiotic use, nutritional support, early enteral nutri- tion and control of blood glucose should be taken into account after the operation of gastrointestinal neoplasm in order to prevent fungal infections.