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.

Glucose Transport Proteins, Facilitative ; deficiency ; Humans ; Syndrome

Adult ; Bone Plates ; Female ; Humans ; Male ; Middle Aged ; Pubic Symphysis Diastasis ; surgery

OBJECTIVEObserve the effects of Goutengsan on SOD, MAO-B, GSH-PX, NO, LDH, index of brain, rate of death and so on in rats to study therapeutic effects and mechanism of Goutengsan on Alzheimer dementia (AD) model.

METHODOne hundred and twenty rats were randomly divided into 6 groups, 3 experimental groups of which were daily administrated with Goutengsan extract whereas the model and control groups were given NS (0.01 mL x g(-1)). Aniracetam at 0.1 g x kg(-1) served as a positive control. At the 5th day after administration, all groups except the control were administrated (ip) with AlCl3 (100 mg x kg(-1) ) for successive 50 days at 1 day interval. After administration, the death rate, body weight, training scores, brain index, MAO-B, SOD, GSH-Px in brain and NO, LDH in serum were determined.

RESULTThe brain index, SOD, GSH-Px activities as well as NO content of drug-treated groups were strikingly higher that of model group, and had not obvious difference from that of normal group except content of LDH was higher.

CONCLUSIONGoutengsan could increase the brain index, cut down the rate of death, stable increase of body weight, promote the endogenous antioxidant activity, enhance the clearance of lipid peroxide and other metabolic waste, inhibit the MAO-B activity, reduced the leakage of LDH and maintain the content of NO at a normal level. Therefore Goutengsan could protect cells, delay senile, improve symptoms of AD.

Aluminum Compounds ; pharmacology ; Alzheimer Disease ; chemically induced ; drug therapy ; metabolism ; Animals ; Body Weight ; drug effects ; Brain ; drug effects ; metabolism ; Chlorides ; pharmacology ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Female ; Glutathione Peroxidase ; metabolism ; Lipid Peroxidation ; drug effects ; Male ; Malondialdehyde ; metabolism ; Memory ; drug effects ; Nitric Oxide ; metabolism ; Oxidative Stress ; drug effects ; Pyrrolidinones ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

To screen the active fractions with lithagogue effects of Pyrrosia lingua from Guizhou province and preliminarily investigate its mechanism. The rats were fed with 1% ethylene glycol and 2% ammonium chloride to establish the nephrolithiasis models, which were used to evaluate thelithagogue effects of different polar fractions of P. lingua from Guizhou province. The level of urinary calcium and oxalic acid in urine, renal calcium, oxalic acid, superoxide dismutase (SOD), catalase(CAT) and malondialdehyde (MDA) in renal tissues,as well as crystalline deposit and lithogenesis in renal tissues and the levels of creatinine(Cr) and blood urea nitrogen (BUN) in the serum were detected. The effective compounds were inferred from the analysis of active fractions extract based on LC-MS technology. Petroleum ether fraction and dichloromethane fraction of P. lingua from Guizhou province can reduce renal oxalic acid and renal calcium concentration, increase urinary oxalic acid and urine calcium, with significant inhibitory effect on the formation of renal calculus in rats, significantly increase SOD and CAT activities in renal tissues, and significantly reduce MDA levels. LC-MS analysis showed that the caffeine, citric acid and tartaric acid among the compounds from petroleum ether fraction and dichloromethane fraction had lithagogue effects. Both the petroleum ether fraction and dichloromethane fraction of P. lingua from Guizhou province showed good effect on prevention and treatment of calculus in middle dose groups, and the mechanism may be associated with antioxidation, reducing calcium oxalate crystal deposition, and promoting calcium oxalatecrystal release, in addition, caffeine, citric acid and tartaric acid had lithagogue effects.

OBJECTIVETo investigate the way of resection of high-sacrum tumors and the way and duration of the spinal-pelvic TSRH or ISOLA internal fixation.

METHODFrom October 1998 through April 2002, 35 patients with sacral tumor were enrolled in our hospital, including 4 cases in L(5)-S(1), 2 in L(5)-S(2), 4 in S(1), 8 in S(1 - 2), 6 in S(1 - 3), 6 in S(1 - 4), 5 in S(1 - 5). 35 patients were followed by lumbo-pelvic TSRH or ISOLA internal fixation and corresponding chemotherapy and radiotherapy.

RESULTSIn the follow-up period of 6 - 42 months, the short-term results were satisfactory with the lumbosacral pain reduced and the neurological function improved in different degrees, however dysuria occurred in 1 case and skin necrobiosis at coccygeal incision occurred in 1 case; two cases experienced cerebrospinal fluid leakage and 1 case experienced postoperative infection and delayed healing, 1 case with chordoma and 2 cases with malignant fibrous histiocytoma recurred 1 year after postoperation, one of these 2 cases with malignant fibrous histiocytoma suffered from lung metastasis and died of system failure 19 months after postoperation. No fractured rod occurred.

CONCLUSIONSurgical procedure and postoperative comprehensive treatment have important effects on the prognosis. High-sacral tumor resection and reconstruction are effective means of achieving stabilization, providing significant pain relief and preserving ambulatory capacity.

Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; Male ; Middle Aged ; Pelvic Bones ; surgery ; Sacrum ; Spinal Neoplasms ; surgery ; Treatment Outcome ; Young Adult

Adult ; Female ; Humans ; Menstruation ; physiology ; Middle Aged ; Occupational Health ; Stress, Psychological ; epidemiology ; Surveys and Questionnaires

BACKGROUNDNanobone putty is an injectable and bioresorbable bone substitute. The neutral-pH putty resembles hard bone tissue, does not contain polymers or plasticizers, and is self-setting and nearly isothermic, properties which are helpful for the adhesion, proliferation, and function of bone cells. The aim of this study was to investigate the osteogenic potential of human bone morphogenetic protein 2 (hBMP2) gene activated nanobone putty in inducing ectopic bone formation, and the effects of the hBMP2 gene activated nanobone putty on repairing bone defects.

METHODSTwenty four Kunming mice were randomly divided into two groups. The nanobone putty + hBMP2 plasmid was injected into the right thigh muscle pouches of the mice (experiment side). The nanobone putty + blank plasmid or nanobone putty was injected into the left thigh muscle pouches of the group 1 (control side 1) or group 2 (control side 2), respectively. The effects of ectopic bone formation were evaluated by radiography, histology, and molecular biology analysis at 2 and 4 weeks after operation. Bilateral 15 mm radial defects were made in forty-eight rabbits. These rabbits were randomly divided into three groups: Group A, nanobone putty + hBMP2 plasmid; Group B, putty + blank plasmid; Group C, nanobone putty only. Six rabbits with left radial defects served as blank controls. The effect of bone repairing was evaluated by radiography, histology, molecular biology, and biomechanical analysis at 4, 8, and 12 weeks after operation.

RESULTSThe tissue from the experimental side of the mice expressed hBMP2. Obvious cartilage and island-distributed immature bone formation in implants of the experiment side were observed at 2 weeks after operation, and massive mature bone observed at 4 weeks. No bone formation was observed in the control side of the mice. The ALP activity in the experiment side of the mice was higher than that in the control side. The tissue of Group A rabbits expressed hBMP2 protein and higher ALP level. The new bone formation rate and antibending strength of group A was significantly higher than those of group B and C. The defects in blank control were not healed.

CONCLUSIONSThe hBMP2 gene activated nanobone putty exhibited osteoinductive ability, and had a better bone defect repair capability than that of nanobone putty only.

Absorbable Implants ; Animals ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; analysis ; genetics ; Female ; Male ; Mice ; Osteogenesis ; physiology ; Rabbits ; Transforming Growth Factor beta ; analysis ; genetics

OBJECTIVE Bingpian is an almost pure chemical with a chemical composition of (+)-borneol and has been historically used as a topical analgesic in traditional Chinese medicine for millen-nia. However, the clinical efficacy of topical bingpian lacks stringent evidence-based clinical studies and the underlying molecular mechanisms are unclear.This study verified the analgesic efficacy of topi-cal bingpian in humans, and elucidated the underling mechanisms in animal models of pain. METH-ODS The analgesic efficacy of topical bingpian was examined in a randomized,double-blind,placebo-controlled clinical study at the Shanghai Changzheng Hospital. Capsaicin, formalin, CFA or thermal caused pain/hyperalgesia were established in different mouse models,and bingpian-induced analgesia and the underlying mechanisms were studied in these models.The molecular targets of bingpian were examined by calcium imaging, patch-clamp recording and enzymatic activity assay in mouse sensory neurons or transfected HEK 293 cells. RESULTS (1)Topical application of bingpian leads to significantly greater pain relief than placebo does in a randomized, double-blind, placebo-controlled clinical study involving 122 patients with postoperative pain.(2)TRPM8 channel is the most sensitive molecular target of bingpian and mediates topical bingpian-induced analgesia in mice. (3)A downstream glutamatergic mechanism in the spinal cord contributes to topical bingpian-induced analgesia. (4)Bingpian shows mechanistic differences and advantages as a topical analgesic when compared with menthol.

OBJECTIVEWe retrospectively analyzed the cause and death risk of 114 postoperative respiratory failure patients found in 3519 patients with esophageal cancer and 1495 patients with carcinoma of gastric cardia surgically treated between January 1992 and May 2003.

METHODSTo analyze the reasons causing postoperative respiratory failure in surgically treated esophageal or gastric cardia cancer patients, and the correlation between the death risk of postoperative respiratory failure and preoperative pulmonary function tests, postoperative complications, operation modes, history of preoperative accompanying diseases and so on using Binary Logistic Regression analysis and Chi-square tests (chi(2)) in SSPS statistics software.

RESULTSIn this series, postoperative respiratory failure developed in 97 of 3519 (2.76%) esophageal cancer patients and 17 of 1495 (1.14%) gastric cardia cancer patients, which were mainly caused by severe respiratory tract infection (37.7%, 43/114) and operative complications (35.1%, 40/114) such as: anastomotic leakage or perforation of thoracic stomach, extensive bleeding during operation, chylothorax, etc, totally accounting for 72.8% (83/114). In contrast with lung cancer patients, most of the postoperative respiratory failure (69.3%) occurred in the patients who had perioperative complications but almost always normal preoperative pulmonary function tests. Other reasons to cause postoperative respiratory failure were: extubation in unconscious patients at the end of general anesthesia; over-infusion during operation; pulmonary artery embolism; severe arrhythmia and so on. All patients except 2 were treated in ICU by mechanic ventilation through intubation and/or tracheotomy. Eighty patients (70.2%) were intubated and/or had tracheotomy within 3 days postoperatively. Seventy patients (61.4%) were rescued successfully, whereas 44 cases (38.6%) died of postoperative respiratory failure and/or other postoperative complications. Univariate analysis and multivariate analysis by binary logistic regression indicated that: severe perioperative complications, more postoperative complications, poor preoperative pulmonary function, radical preoperative radiotherapy, intubation and/or tracheotomy after the second postoperative day and long period of mechanic ventilation were the major risk factors leading to death once the postoperative respiratory failure developed. The former 3 factors were independent risk factors leading to death with OR of 2.50, 2.37, 1.68, respectively. Age, sex, operation modes, history of preoperative accompanying disease, prophylactic antibiotics were not demonstrated as statistically significant risk factors correlated with death.

CONCLUSIONSevere perioperative complications and respiratory tract infection are the two major causes of postoperative respiratory failure in patients with cancer of esophagus and gastric cardia. Patients with severe perioperative complications or poor preoperative pulmonary function or association with more than two kinds of postoperative complications have much higher death risk than other patients when they develop postoperative respiratory failure. Careful manipulation during operation and effective perioperative management are the most important measures to avoid postoperative respiratory failure and high mortality.

Adult ; Aged ; Aged, 80 and over ; Cardia ; China ; epidemiology ; Esophageal Neoplasms ; physiopathology ; surgery ; Esophagectomy ; adverse effects ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Postoperative Complications ; etiology ; prevention & control ; Respiratory Function Tests ; Respiratory Insufficiency ; epidemiology ; etiology ; prevention & control ; Retrospective Studies ; Risk Factors ; Stomach Neoplasms ; physiopathology ; surgery