A possible ?-Galactosidase gene(pwtsA) was discovered from soil metagenomic DNA of Taishan Mountain.PwtsA gene was inserted into the expression vector pET30a and transferred into E.coli BL21(DE3).Recombinant protein PWTSA was expressed as a soluble form at high level through IPTG induction,with a molecular mass of 57 kD analyzed by SDS-PAGE.PWTSA can produce o-nitrophenol from o-nitrophenol-?-D-galactopyranoside(ONPG),and its specific activity was determined as 13.6 U/mg.The enzymatic studies demonstrated that the recombinant protein PWTSA was a thermostable ?-Galactosidase,its optimum temperature and pH were 85?C～95?C and 6.5 respectively.In standard assays,the Km for ONPG was 0.83 mmol/L.

Objective To investigate the incidence of osteoporosis (OP) in patients with ankylosing spondylitis and the relationship between OP and the clinical data. Methods Serum levels of bone alkaline phosphatase (BALP) and tartrate resistant acid phosphatase 5b (TRACP5b) were detected by enzyme linked immunosorbent assay ( ELISA ) in 60 cases with ankylosing spondylitis, and it was compared with normal controls. Bone mineral density (BMD) was measured through dual-energy X-ray absorptiometry ( DXA), including lumbar ( L2 - L4), bilateral femoral neck and greater trochanter. Some clinical data was collected and analyzed at the same time. Results The incidence of OP in AS patients was 35%, and the incidence of OP in the femoral proximal end was higher than that in lumbar. Compared with normal controls[ ( 1.06 ±0. 18 )U/L ], the levels of serum TRACP5b in AS[ (1.31 ± 0. 82 )U/L] patients was significantly higher ( P <0. 05 ). The levels of serum BLAP in OP combined AS group[ ( 21.65 ± 5.41 ) U/L]were significantly lower than non-OP group[ (32. 37 ± 16. 5 ) U/L] ( P <0. 05 ). The disease duration was negatively correlated with the BMD of femoral neck ( P < 0. 01 ). Conclusions There was higher incidence of OP in AS patients, which were related with the abnormality of bone metabolism and the disease duration.Multiple factors participated in the regulation of bone metabolism of AS.

Adenocarcinoma ; complications ; pathology ; Aged ; Humans ; Male ; Neoplasms, Multiple Primary ; complications ; pathology ; physiopathology

Objective To evaluate the effects of a health education project on endemic fluorosis in Shandong Province,and to provide a basis for formulating control strategies.Methods From December 2010 to June 2011,according to historical conditions,a total of 19 counties (cities,districts) of Shandong Province were chosen,and 3 townships (towns) were chosen in each project county.Health educational activities on endemic fluorosis were carried out in the Central Primary School in grade 4 to 6 in each township(town).In each project township(town),3 villages were chosen in each selected township(town) where the health educational activities in the community were carried out.Before and after the health educational activities,surveys on knowledge questionnaire on drinking-water-borne fluorosis control were conducted among 30 students of grade 5 in the Central Primary School and 15 housewives in every school location in each selected township(town).Results After the health educational activities,the knowledge awareness rates of endemic fluorosis control of the students and housewives were 96.53％ (5482/5679) and 94.88％ (3501/3690),respectively,and increased significantly compared with those before intervention [62.31％ (5154/8271) and 76.91％ (2815/3660)],and the difference was statistically significant (x2 =2176.50,490.58,all P ＜ 0.01).Among the primary school students and housewives,the knowledge awareness rates of endemic fluorosis control were increased by 34.22％ and 17.97％,respectively.Conclusions Health education activities on endemic fluorosis can significantly improve the knowledge awareness of target population,which will play a positive role in promoting prevention and control of endemic fluorosis.

Objective To observe the influence of nandrolone phenylpropionate(NP) on the ultrastructure of aorta in rats with or without movement training,and investigate the side effects of NP on the cardiovascular system. Methods Forty male SD rats were randomly divided into sedentary control group,sedentary+medicine group,exercise control group and exercise+medicine group.For the groups with medical treatment,NP of 10 mg/kg one time every three days was injected into the rats via gluteus for eight weeks.For the exercise groups,rats were trained to run on treadmill five days per week for eight weeks.The aortae were sampled and specimens were obtained for transmission electron microscopy. Results The ultrastructure of aorta was normal in sedentary control group.For sedentary+medicine group,mitochondrial swelling,vacuolated cytoplasm and lysis of endothelial cells were observed,disruption of intercellular conjunctions,widening of subendothelial spaces and furcation and breakage of internal elastic lamina were found,and smooth muscle cells changed into synthesis type.For exercise control group,no obvious morphologic change was observed,except that part of the internal elastic lamina disrupted.In exercise+medicine group,breakage and lysis of endothelial cells were observed,widening of subendothelial spaces and lysis of internal elastic lamina were found,and autophagosome and myelinoid body were seen in smooth muscle cells. Conclusion NP may lead to the impairment of endothelial cells and the change of smooth muscle cells into synthesis type.Exercise with NP administration may result in more severe impairment in vessel wall.

Objective To investigate the change of the neutrophil apoptosis and neutrophil respiratory burst in the patients and the effect of ulinastatin on the apoptosis of neutrophil and respiratory burst of neutrophil during cardiopulmonary bypass (CPB). Methods Sixty-two patients undergoing valve replacement with CPB were randomly divided into two groups: ulinastatin group (U group, 31 cases) and control group (C group, 31 cases). In U group patients received ulinastatin after induction of anesthesia. In C group patients received equal volume of normal saline, instead of ulinastatin. Arterial blood was obtained before operation (T1), 30 min after the start of CPB (T2), 30 min after the termination of CPB (T3). The apoptosis of neutrophil and respiratory burst of neutrophil were measured by flow cytometer. The level of superoxide dismutase (SOD) and malondialdehyde (MDA) were measured by kit. Results In C group, compared with T1 [(66.57±5.93)%], the rate of the apoptosis of neutrophil was significantly decreased at T2[(55.37±3.51)%] and T3 [(48.92±4.21)%] (P<0.05). And in U group, compared with T1 [(73.57±7.94)%], the rate of the apoptosis of neutrophil was significantly decreased at T2 [(68.34±4.92)% ] and T3 [(62.13±4.76)%] (P<0.05), And it reached to the minimum at T3. The rate of the neutrephil apeptosis was significantly lower in C group than that in U group (P<0.05). The respiratory burst of neutrophil increased significantly after the start of CPB and reached to the peak at T3[C group (1422.50±89.75) MCF,U group (1156.52±93.20) MCF]. The respiratory burst of neutrophil in U group was significandy lower than that in C group at T2 and T3 (P<0.05). The vitality of SOD decreased significantly after the start of operation in the two groups (P<0.05). The level of MDA increased significantly after the start of operation in the two groups, and reached to the peak at T3. The vitality of SOD in C group was significantly lower than that in U group at T3 (P<0.05). The level of MDA in C group was significantly higher than that in U group at T3 (P<0.05). Conclusions The rate of neutrophil apoptosis decreased and the respiratory burst of neutrophil increased during CPB. By improving the apoptosis of neutrophil and reducing the respiratory burst of neutrophil, ulinastatin can inhibit inflammatory reaction during CPB. Meanwhile, ulinastatin can improve the vitality of SOD and reduce the level of MDA. In conclusion, ulinastatin has a significant protective effect during CPB.

Bronchogenic Cyst ; pathology ; surgery ; Diagnosis, Differential ; Gastrectomy ; methods ; Humans ; Male ; Middle Aged ; Stomach Diseases ; pathology ; surgery

Objective To determine whether the anti-inflammatory properties of Shuxiong Tablet(SXT)and the effective components group of SXT(ECGS)are equivalent and to assess the formulary rationality.Methods ECGS consisted of Panax notoginsen saponion(PNS),hydroxysafflor yellow A,and ferulic acid plus volatile oil of Ligusticum chuanxiong,which was based on the active ingredients and their ratios in SXT.We compared the anti-inflammatory actions of ECGS and SXT using the xylene-induced edema model and the carrageenan-induced edema model,as well as the analgesic activity of them using the acetic acid-induced writhing model.Moreover,cultured macrophages were incubated with media containing serum isolated from SXT-,ECGS-,or every component of ECGS-treated rats,to compare the depress effects on lipopolysaccharide(LPS)-stimulated NO production and inducible nitric oxide synthase(iNOS)expression.Results ECGS and SXT had equivalent anti-inflammatory actions and analgesic effects at an equipotent dosage in a dose-dependent manner.The drug-containing media could inhibit the LPS-stimulated NO production and iNOS expression in cultured macrophages.A 2 × 2 × 2 ANOVA revealed that three effective components could produce synergistic effect on the inhibition of NO production,and PNS was the capital component.Conclusion ECGS and SXT display an equivalent anti-inflammatory effect,and the formula follows traditional Chinese medicine compatibility principle,which shows obvious formulary rationality.

BACKGROUNDArthritogenic T lymphocytes with common T cell receptor (TCR) Vbeta clonotypes, infiltrating in the articulars of rheumatoid arthritis (RA) patients, play a central role in the pathogenesis of RA. TCR Vbeta5.2 and TCR Vbeta8.2 are the main pathogenic T cell clonotypes in the course of collagen-induced arthritis (CIA) progression in Lewis rats. To investigate a TCR-based immunotherapy for RA, we constructed recombinant DNA vaccines encoding TCR Vbeta5.2 and TCR Vbeta8.2, and evaluated the inhibitive effects of the two vaccines on CIA rats.

METHODSGenes encoding TCR Vbeta5.2 and TCR Vbeta8.2 were amplified by RT-PCR from spleen lymphocytes of Lewis rats and cloned into the eukaryotic expression vector pTargeT. The expression of vaccines was confirmed by RT-PCR and immunohistochemistry. The inhibitive effects of the vaccines on articulars of CIA rats were assessed with arthritis index evaluation and histology. Interferon gamma (IFN-gamma) and interleukin (IL)-4 production by spleen lymphocytes were tested with enzyme-linked immunospot assay (ELISPOT) technique, the changes in peripheral CD4(+) and CD8(+) lymphocyte populations were tested by flow cytometry, and the level of anti-CII antibody in serum was assayed by enzyme-linked immunosorbent assay (ELISA).

RESULTSRecombinant DNA vaccines pTargeT-TCR Vbeta5.2 and pTargeT-pTCR Vbeta8.2 were successfully constructed. Both vaccines inhibited CIA, which alleviated the arthritis index score (P < 0.05), decreased the level of IFN-gamma (P < 0.05), and reduced the ratio of CD4(+)/CD8(+) lymphocytes (P < 0.05) and the anti-CII antibody in serum (P < 0.05). In addition, the histological change in DNA-vaccinated rats was less serious than CIA rats. Compared to pTCR Vbeta 8.2 and pTCR Vbeta 5.2 groups, the group that was injected with a combination of the two vaccines showed stronger inhibitive effects on CIA than either individual vaccine.

CONCLUSIONThe recombinant plasmids pTargeT-TCR Vbeta5.2 and pTargeT-TCR Vbeta8.2 have obvious inhibatory effects on CIA rats and better effects could be achieved when the vaccines were used in combination.

Animals ; Arthritis, Experimental ; metabolism ; prevention & control ; CD4-Positive T-Lymphocytes ; drug effects ; CD8-Positive T-Lymphocytes ; drug effects ; Enzyme-Linked Immunosorbent Assay ; Female ; Flow Cytometry ; Immunohistochemistry ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Muscles ; drug effects ; metabolism ; Peptide Fragments ; antagonists & inhibitors ; Rats ; Rats, Inbred Lew ; Receptors, Antigen, T-Cell, alpha-beta ; antagonists & inhibitors ; Reverse Transcriptase Polymerase Chain Reaction ; Vaccines, DNA ; pharmacology

OBJECTIVETo study the rehabilitation effects ergometry on COPD patients.

METHODSThirty COPD out-patients in our Hospital were randomly divided into 2 groups. Rehabilitation group, 15 patients, performed leg ergometry exercise of 80% peak Watt x 30min/d x 3d/w x 12w. Another 15 patients were control group without exercise. All patients received conventional therapy. Pulmonary function testing (PFT), cardiopulmonary exercise testing (CPET), arterial blood gas analysis (ABG), Borg and CAT sores were done at both baseline and 12 w.

RESULTSThere was no statistically difference in lung function testing, blood gas analysis and cardiopulmonary exercise test when pre- exercises between 2 sub-groups. The IC, peak VO2 and peak, W of rehabilitation group significantly increased (P < 0.05); and Borg and CAT.scores significantly decreased (P < 0.05) from baseline; and other PFT and ABG did not change (P > 0.05). While there was no difference in control group (P > 0.05).

CONCLUSIONLeg submaximal ergometry rehabilitation improves health condition and ameliorate dyspnea symptoms in COPD patients.

Blood Gas Analysis ; Dyspnea ; therapy ; Exercise Test ; Exercise Therapy ; Humans ; Pulmonary Disease, Chronic Obstructive ; therapy ; Respiratory Function Tests