In order to improve the thermostability of the Penicillium expansum Lipase (PEL), the lipase encoding genes was mutated by site-directed mutagenesis. A recombinant vector pAO815-ep8-K55R which contain double mutant genes was constructed by overlap extension PCR using the cDNA of a random-mutant lipase ep8 (a single site mutant) as the template and two special primers were used to generate another mutation site K55R. The recombinant vector was transformed into Pichia pastoris GS115 by electroporation and the recombinant mutant GS-pAO815-ep8- K55R can secret double-mutant lipase PEL-ep8-K55R-GS into the medium when it was induced by Methanol. The yield of the double-mutant lipase is 508 u/mL, which is 81% that of the wild type lipase PEL-GS (627 u/mL) and 55% that of random-mutant PEL-ep8-GS (924 u/mL). The specific activity of double-mutant lipase is 2309.1 u/mg, which is similar to random-mutant lipase PEL-ep8-GS and the wild type lipase PEL-GS. The optimum temperature of the double-mutant lipase is same with the wild type lipase PEL-GS and random-mutant lipase PEL-ep8-GS. While the Tm of the double-mutant lipase is 41.0 degrees C, 2.3 degrees C higher than the wild type lipase PEL-GS and 0.8% higher than the random-mutant lipase PEL-ep8-GS, indicating that the double-mutant lipase PEL-ep8-K55R-GS has higher thermostability.
Electroporation ; Enzyme Stability ; Hot Temperature ; Lipase ; genetics ; metabolism ; Mutagenesis, Site-Directed ; Mutant Proteins ; metabolism ; Penicillium ; enzymology ; Pichia ; genetics ; metabolism ; Protein Engineering ; methods ; Recombinant Proteins ; biosynthesis ; genetics

OBJECTIVETo observe the clinical efficacy difference between segmentation scraping and conventional acupuncture for cervical spondylosis (CS) so as to provide effective treatment method.

METHODSEighty-five cases of cervical type of CS were randomly divided into a scraping group (44 cases) and an acupuncture group (41 cases). The segmentation scraping therapy was used in the scraping group. The scraping group was treated with focusing on scraping the head and joint part of neck and occiput in the upper cervical spine injury, and focusing on scraping the lower section of cervical and shoulder in the lower cervical spine injury, once every seven days, totally for 3 times. In the acupuncture group, Fengchi (GB 20),Wangu (TE 5), Tianzhu (BL 10),Neck-Jiaji (EX-B 2), etc. were selected,once daily,for 15 days. The visual analogue scale (VAS) was used to evaluate the immediate analgesic effect after the first treatment and the clinical efficacy was observed after the end of treatment.

RESULTSAfter the first treatment, the score of VAS was decreased significantly in the scaping group (P < 0.01), but there was no significant difference in the acupuncture group compared with those before treatment (P > 0.05); the score of VAS in the scaping group after the first treatment was lower than that in the acupuncture group (3.66 +/- 0.74 vs 5.43 +/- 0.35, P < 0.01). Compared with before treatment, the scores of VAS were decreased significantly after treatment in two groups (both P < 0.01), but without significant difference between two groups (P > 0.05); the effective rate was 95.5% (42/44) in the scaping group and 87.8% (36/41) in the acupuncture group, the curative effects were similar (P > 0.05).

CONCLUSIONBoth of scraping and acupuncture therapies have good analgesic effect for cervical spondylosis, and overall effects are similar, but the immediate analgesic effect of scraping thrapy is better than that of conventional acupuncture.

Acupuncture Points ; Acupuncture Therapy ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Neck Pain ; therapy ; Spondylosis ; therapy ; Treatment Outcome

OBJECTIVETo assess the efficacy and safety of recombinant human thrombopoietin (rhTPO) on chemotherapy-induced thrombocytopenia in patients with solid tumor.

METHODSIn this randomized crossover self-controlled multi-center clinical trial, 154 patients with solid tumor were randomly divided into two groups (group A 77 cases and group B 77 cases). All patients were given the same two cycles of chemotherapy. In group A, the first cycle was treated cycle, in which patients were given rhTPO, while the second cycle was non-treated cycle as a control. In group B, the first cycle was non-treated cycle as a control, while the second cycle was treated cycle. RhTPO 1.0 microg/(kg x d) was administered subcutaneously 6-24 hours after chemotherapy for the longest 14 days. Laboratory tests included complete blood counts, urinalysis, serum chemistry, coagulant test, chest radiography, and electrocardiogram. Serum samples were screened for anti-rhTPO antibodies.

RESULTSIn both group A and group B, platelet decrease and duration had no significant difference between the treated cycle and non-treated cycle. Platelet count was higher in the treated cycle, than in the non-treated cycle: [minimal mean platelet count (64.4 +/- 45.4) x 10(9) cells/L and (52.4 +/- 30.9) x 10(9) cells/L (P=0.000), maximal mean platelet count (263.9 +/- 142.5) x 10(9) cells/L and (148.9 +/- 67.7) x 10(9) cells/L (P=0.000)]. Duration of thrombocytopenia was shorter in the treated cycle than in the non-treated cycle [days with platelet count < 50 x 10(9) cells/L, (2.5 +/- 3.9) and (3.7 +/- 5.7) (P=0.04); days with platelet count recovered > or = 75 x 10(9) cells/L, (10.3 +/- 8.7) and (14.0 +/- 8.9) (P=0.000), and days with platelet count recovered > or = 100 x 10(9) cells/L, (15.9 +/- 10.5) and (21.1 +/- 9.5) (P=0.000)]. The need for platelet transfusion was not significantly reduced in treated cycle. The effects of rhTPO on WBC, Hb, hepatic function, renal function, and coagulant function were not found. Transient low-titer non-neutralizing antibody was developed in one patient. Therapy with rhTPO was tolerated by all patients. Mild side effects were observed in individual patients, including fever, dizziness, and chill. Conclusion Administration of rhTPO after chemotherapy can significantly reduce the degree and duration of thrombocytopenia and promote platelet recovery. Therapy with rhTPO seems to be safe.

Adolescent ; Adult ; Aged ; Antineoplastic Agents ; adverse effects ; Cross-Over Studies ; Female ; Humans ; Lung Neoplasms ; blood ; drug therapy ; Male ; Middle Aged ; Neoplasms ; blood ; drug therapy ; Platelet Count ; Recombinant Proteins ; administration & dosage ; therapeutic use ; Thrombocytopenia ; chemically induced ; drug therapy ; prevention & control ; Thrombopoietin ; therapeutic use

OBJECTIVETo analyze the distribution features of systemic vascular resistance (SVR) disorder in healthy subjects in Hebei province.

METHODSSVR and systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), pulse pressure (PP), heart rate (HR), cardiac output (CO), cardiac index (CI), stroke volume (SV), stroke index (SI), left ventricular ejection time (LVET), left cardiac work (LCW) and cardiovascular function were determined with Bioz. com Cardio Dynamics. Total cholesterol (TC), triglyceride (TG) and influencing factor were tested by automated hematology analyzer in 999 healthy subjects living in Hebei province.

RESULTSSVR disorder gradually increased with the ageing in healthy subjects and the prevalences of SVR disorders was higher among females than among males (P < 0.001). SVR was positively correlated with SBP, DBP, MAP, PP, LVET, body mass index (BMI), TC, and TG, and was inversely associated with HR, CO, CI, SV, SI, LCW, and arterial compliance (AC). CO and MAP were independent influencing factors with SVR.

CONCLUSIONSVR disorder is associated with age, sex, and blood pressure in populations in Hebei province and may be a marker of vascular injury.

Adolescent ; Adult ; Age Factors ; Aged ; Blood Pressure ; Body Mass Index ; Cardiac Output ; Female ; Hemodynamics ; physiology ; Humans ; Male ; Middle Aged ; Risk Factors ; Sex Factors ; Vascular Capacitance ; Vascular Diseases ; physiopathology ; Vascular Resistance

Objective To evaluate diagnosis and preoperative management of cases with interrupted aortic arch(IAA)in infancy.Methods Fifty-three infants who were admitted to our hospital from Jan.2001 to Nov.2007 were involved.Clinical data,findings of echocardiogram(Echo),spiral CT,MRI,angiocardiography,preoperative management,surgical repair and postoperative outcome were analyzed retrospectively.Results There were 38 boys and 15 girls,weighted 2.18-10.0(4.32?1.60)kg,aged 1 day to 12(3.05?3.53)months,of which 50.94% were neonates.Symptoms at presentation were 90.57% with tachypnea and 83.02% with difficulty in feeding.Eighty three point zero two percentage cases had different degree of congestive heart failure,37.74% of which were in grade Ⅲ heart function.All cases had weakened femoral pulse.All cases were performed Echo,38 cases of them diagnosed as IAA,6 cases as IAA or severe coarctation of aorta(CoA);they were diagnosed as IAA by CT,and 9 as severe CoA who were diagnosed as IAA via CT or operation.Thirty-three cases were performed CT,of which 15 underwent surgical repair,cardiovascular abnormalities revealed by CT were the same as those in surgical findings.Three cases were taken MRI,and 7 cases were performed angiocardiography.According to the results of Echo,CT,MRI,angiocardiography and surgical findings,35 cases were type A,15 cases were type B and 3 cases were type C.Preoperative treatment included maintaining patent ductus areriosis,management of heart failure and supportive treatment.After proper preoperative management of medication,most cases with congestive heart failure were improved.Twenty-six cases underwent surgical repair,16 survived,10 died du-ring perioperative stage.Main cause of death was severe low cardiac output.Conclusions Value of Echo in diagnosis of IAA is limi-ted.Combination of Echo with CT or MRI is a convenient and safe way to diagnose IAA,it can replace the traditional method of Echo combined with angiocardiography.Proper preoperative management is helpful to patients with IAA to pass to surgical repair,and makes for successful operation.

OBJECTIVETo investigate whether age influence the precision of dual X-ray absorptiometry (DXA) measurement at the hip in middle-aged and elderly women.

METHODSTotally 90 women were randomly selected and divided into three age groups: 45-55 years, 56-65 years, and 66-75 years. Each age group contained 30 women. Each woman was scanned twice at the same day. Bone mineral density (BMD) values of femoral neck, ward's triangle, and trochanter were collected and grouped by calculating the root mean square (RMS). Precision errors were expressed as RMS (standard deviation, SD).

RESULTSFor the femoral neck and trochanter, significant differences of SD of BMD existed among all age groups. For the ward's triangle, significant difference of BMD existed among all age groups except between the 45-55 group and 56-65 age group.

CONCLUSIONAge can influence the precision of DXA measurement at the hip in middle-aged and elderly women.

Absorptiometry, Photon ; Age Factors ; Aged ; Analysis of Variance ; Bone Density ; Female ; Femur ; physiology ; Femur Neck ; physiology ; Humans ; Menopause ; physiology ; Middle Aged ; Reproducibility of Results

OBJECTIVETo investigate the influence of intravenous infusion of 50 g/L fructose on post-operative blood glucose level in burn patients, and to evaluate its therapeutic value and safety.

METHODSA prospective, randomized, double blinded clinical trial was conducted. Forty-one burn patients with burn area ranging between 10% -30% of total body surface (TBSA) and third degree burns ranging between 1% -10% TBSA were enrolled in the study and randomized into experiment group (E, n = 21, with intravenous infusion of 500 ml of 50 g/L fructose daily for 3 days after escharectomy) and control group ( C, n = 20, with intravenous infusion of glucose 1 day after escharectomy for 3 days). Intravenous infusion of other carbohydrate liquids or oral intake of sugar was withhold within 4 hours of fructose or glucose infusion. Physical signs and side effects were observed during the administration. The plasma glucose contents before operation and on 1, 2 and 3 post-operation day( POD) were measured. The serum content of lactic acid, uric acid, hepatic and renal function were determined before operation and on 4 POD.

RESULTSPhysical signs before and after drug administration, and plasma glucose content before operation, as well as before and after fructose administration in 3 POD exhibited no obvious difference between the two groups ( P > 0. 05 ). The plasma glucose content was increased 3 days after operation in the control group, and it reached the peak on 3 POD [ (8. 4+/-3. 5) mmol/L] , which was markedly higher than that before glucose administration [ (6. 4+/-2.4) mmol/L, P <0. 01) ]. The plasma contents of lactic acid and uric acid showed no obvious difference ( P >0.05) between the two groups, and also no difference before and after operation ( P > 0. 05). No changes were observed in hepatic and renal functions.

CONCLUSIONIntravenous infusion of 50 g/L D-fructose is safe because it exerts little influence on blood glucose level.

Adolescent ; Adult ; Aged ; Blood Glucose ; drug effects ; Burns ; blood ; drug therapy ; Double-Blind Method ; Female ; Fructose ; adverse effects ; therapeutic use ; Humans ; Infusions, Intravenous ; Male ; Middle Aged ; Postoperative Period ; Prospective Studies

Amyloid β protein (Aβ) is closely involved in the pathogenesis of Alzheimer's disease (AD), and one of the main strategies for AD treatment is antagonizing the neurotoxicity of Aβ or even clearing the Aβ deposited in the brain. The present study was aimed to observe the effects of intrahippocampal injection of Aβ₃₁₋₃₅ on the spatial learning and memory of rats by using Morris water maze technique, and explore the neuroprotective effects and possible mechanism of [Gly14]-humanin (HNG) against Aβ-induced deficits in learning behavior. The results showed that bilateral intrahippocampal injection of 2.0 nmol Aβ₃₁₋₃₅ significantly increased the mean traveled distance of rats in searching for the hidden underwater platform and decreased the distance percentage in the target quadrant in probe test after withdrawal of platform, whereas pretreatment with HNG (0.2 nmol and 2.0 nmol) suppressed Aβ₃₁₋₃₅-induced increase in the traveled distance and decrease in swimming distance percentage. Application of Genistein (40 nmol), a specific tyrosine kinase inhibitor, almost completely blocked the antagonistic effects of HNG against Aβ₃₁₋₃₅. These results indicate that HNG can dose-dependently prevent against Aβ₃₁₋₃₅-induced impairment in spatial learning and memory of rats, and the neuroprotective effects of HNG might involve the activation of endogenous tyrosine kinase pathway, suggesting that up-regulation of the tyrosine kinase signaling by using HNG might be of great significance for the improvement of cognitive function in AD.
Alzheimer Disease ; physiopathology ; Amyloid beta-Peptides ; adverse effects ; antagonists & inhibitors ; Animals ; Brain ; drug effects ; Genistein ; pharmacology ; Memory ; drug effects ; Neuroprotective Agents ; pharmacology ; Peptide Fragments ; adverse effects ; antagonists & inhibitors ; Peptides ; pharmacology ; Rats ; Spatial Learning ; drug effects

OBJECTIVETo study the relationship between ultrasonic bone mineral density (BMD) of calcaneus and age, height, and body weight, and to establish the normal reference value for stiffness index (SI) of healthy subjects in Guangxi autonomous region and provide scientific foundation for the diagnosis and prevention of osteoporosis.

METHODSSI of calcaneus in 1 816 healthy adults aged 11-90 years (775 men, 1 041 women) were measured with quantitative ultrasound. According to their ages, all the men and women were divided into 13 groups by 5-year age group.

RESULTSSI reached peak values in 21-25 year group in men (range: 111.45 +/- 21.19) and in 16-20 year group in women (range: 101.26 +/- 17.51). Then the SI value began to decline with aging. The SI featured by a typical dual-peak curve in women and the decrease rate of SI was faster in women than in men over 61 years. Multivariate linear regression analysis showed significantly negative correlation between SI and age (P < 0.001), positive correlation between SI and body weight (P < 0.05 for men; P < 0.01 for women), and positive correlation between SI and height in women (P < 0.01).

CONCLUSIONSI correlates with age, height, and weight. The acquired SI value may provide a reference for the diagnosis of osteoporosis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Density ; Calcaneus ; diagnostic imaging ; Child ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Osteoporosis ; diagnosis ; epidemiology ; Ultrasonography

OBJECTIVETo investigate the lung function of the normal subjects living in Hebei province and its correlative factors such as living circumstance, age, height, and body weight.

METHODSThe lung volumes and breath capacities of 1,587 normal subjects were tested by portable spirometers (Scope Rotry) from August to October in 2002. The influences of living circumstance, age, gender, height, and body weight on lung functions were observed and analyzed.

RESULTSNo significant difference was found between urban and rural areas in all indexes (P > 0.05); however, significant difference existed between male and female subjects (P = 0.000). The change trends of lung function in male and female subjects were similar. Growth spurt appeared at the age of 12-16 years in male subjects and 12-14 years in female subjects. Vital capacity (VC), forced vital capacity (FVC), and forced expiratory volume in one second (FEV1) reached their peaks at the age of 26-34 years and then decreased with age. Peak expiratory flow (PEF), 25% forced expiratory flow (FEF50%), and 75% forced expiratory flow (FEF75%) appeared at the age of 18 and then went down with age. Both height and weight had a correlation with all the indexes of lung functions, although the influence of height is stronger than weight.

CONCLUSIONSAll the indexes of lung function have correlations with age, height, and weight. Lung function changes with aging, therefore different expected values shall be available for the adolescence, young adults, and middle-aged and old people. This study provides reference values of lung function for normal population.

Adolescent ; Adult ; Aged ; Child ; Female ; Forced Expiratory Flow Rates ; Forced Expiratory Volume ; Humans ; Lung ; physiology ; Male ; Middle Aged ; Peak Expiratory Flow Rate ; Reference Values ; Respiratory Function Tests ; Rural Health ; Sampling Studies ; Urban Health ; Vital Capacity