A simple and rapid expression and purification method of recombinant firefly luciferase was developed for bacteria detection. A modified luciferase gene from North American firefly Photinus pyralis was cloned into pET28a expression vector and the recombinant protein was produced in Escherichia coli BL21. The recombinant luciferase, equipped with a polyhistidine affinity tag, was purified by immobilized metal ion affinity chromatography (IMAC). The approach generated an abundant expression and an efficient purification of a recombinant luciferase with final yield 1.995mg/L of cell culture. Experiments on the recombinant luciferase also showed that the relative light units (RUL) of the enzyme were 5.8×108, and the specific activity was 2.9×1010 RLU/mg. By applying adenosine triphosphate (ATP) bioluminescence to detection of the coin bacteria using the recombinant protein, the ATP content of bacteria was 9.48×10-16mol/mL, and was identical to the bacteria counts (4500CFU/mL) in order of magnitude. Taken together, our results provided a simple and efficacious method of the preparation of recombinant luciferase, which could be applied in the determination of bacteria via ATP bioluminescence.

Adult ; Altitude ; Arteries ; metabolism ; Calcitonin Gene-Related Peptide ; blood ; Humans ; beta-Endorphin ; blood

OBJECTIVETo explore the effects of chronic hypoxia on left and right ventricular function and the expression of cardiac transient receptor potential canonical (TRPC) channels in rats.

METHODSForty eight SD male rats were randomly divided into control group (CON) and chronic hypoxic pulmonary hypertension model group (CH) (n = 24). In CH group, rats were exposed in chronic hypoxia environment (10% +/- 0.2% O2) to induce myocardial hypertrophy. After 3 weeks, mean systemic arterial pressure (mSAP), right ventricular systolic pressure (RVSP), left ventricular systolic pressure (LVSP), left or right ventricular pressure maximum rate of rise (LV/RV + dp/dt(max)), left or right ventricular pressure maximum rate of descent (LV/RV-dp/dt(max)), right ventricular hypertrophy index (RVMI) and left ventricular hypertrophy index (LVMI) were measured. Left and right ventricular myocardium tissue sections were stained by HE and observed under light microscope. Real-time polymerase chain reaction (real-time-PCR) and Western blot were performed to detect the expression of TRPC subfamily.

RESULTSRVSP, RVMI, RV + dp/dt(max) and RV-dp/dt(max) were markedly elevated in CH group (P < 0.01) in comparison to CON group. LVMI was markedly reduced in CH group in comparison to CON group (P < 0.01). LVSP, LV + dp/dt(max) and LV- dp/dt(max) had no significant changes in CH group in comparison to CON group. Right ventricular myocardial cells of CH group became thick, the nuclei stained deeply, the shape of nuclei became not regularity. Left ventricular myocardial fibers did not change significantly. There was significant difference in the levels of mRNA and protein of TRPC1 between CON and CH groups.

CONCLUSIONFor three weeks exposed to chronic hypoxia induced right ventricular hypertrophy specifically, raised the mRNA and protein expression of TRPC1 on right ventricular myocardial cells . TRPC1 might be involved in the development of cardiac hypertrophy.

Animals ; Disease Models, Animal ; Hypertension, Pulmonary ; metabolism ; physiopathology ; Hypoxia ; metabolism ; physiopathology ; Male ; Rats ; Rats, Sprague-Dawley ; Transient Receptor Potential Channels ; metabolism ; Ventricular Function, Left ; physiology ; Ventricular Function, Right ; physiology

The clinicopathological data of 173 cases of inflammatory myofibroblastic tumor (IMT)were retrospectively analyzed.Among them 125 were males and 48 females with a mean age of 47.9 y (5-78 y).The lesions of 117 cases were located in lungs,41 cases in eyes,8 in ileocecum,2 in liver,2 in spleen,1 in abdominal wall,1 in maxillary sinus and 1 in face.The average diameter of lesions were 3.5 cm,ranging from 1.0 cm to 7.0 cm.The clinical manifestations were not specific,depending on the locations of tumor.The imaging examinations were helpful for diagnosis.The prcsence of slender-spindled myofibroblasts and proliferation of fibroblast cells,with infiltration of chronic inflammatory cells were the basic histopathological features of the disease.Combined with the histological characteristics and immunohistochemical staining IMT can be differentiated from other spindle cell tumors.Appropriate surgical resection is the main treatment for IMT.

Humans ; Inservice Training ; Occupational Health ; Safety

Aim To establish a model of the vascular endothelial cell (VEC) injury in SDrats.Methods SD rats were randomly divided into the control and the modelgroups. The model rats were injected with adrenaline diluted to 2. 5 times 0. 05 mg?100 g-1 (tid) for 5 d continously. From the 4th d, they were irritated for 5 min in the0℃ cold-water in the middle between adrenaline injections.The control rats weregiven 0. 9% NS as above. At 6th d, blood samples were taken from carotid arteries ofthe rats and the CEC counts, t - PA、PAI activities, 6-keto-PGF1? concentrations andthe platelet aggregation rate(max) were detected respectively. Results In the modelgroup, as compared with those in the control group, t - PA activity and 6-keto-PGF1?concentration decreased significantly(P

OBJECTIVETo explore the essence of Spleen-Kidney Deficiency in middle-aged patients.

METHODSInvestigation was carried out in 773 cases of 50-69 years old to assay 34 parameters in them, including blood lipid, oxidation and anti-oxidation related substance, sex hormone, liver and renal function, immune function, blood routine, blood pressure and lung vital capacity, etc.

RESULTSSpleen Deficiency Syndrome is closely related with lipid metabolism disorder; Kidney-Qi Deficiency Syndrome is closely related with hypoimmune function; Kidney-Yin Deficiency Syndrome is closely related with lipid metabolism disorder and hypertension, and Kidney Yang Deficiency is closely related with weakness of anti-oxidation capacity, hypoimmunity, the internal environment disorder of sex hormone and aging, manifested as multiple functions abating.

CONCLUSIONSpleen Deficiency, Kidney-Qi Deficiency and Kidney Yang Deficiency are different layers of a gradually developed and aggravated pathological process, but Kidney-Yin Deficiency could not be listed into this layer.

Aged ; Aging ; CD4-CD8 Ratio ; Cholesterol ; blood ; Diagnosis, Differential ; Female ; Humans ; Kidney Diseases ; blood ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Splenic Diseases ; blood ; Superoxide Dismutase ; blood ; Yang Deficiency ; blood ; Yin Deficiency ; blood

Objective To discuss the comparability of blood urea nitrogen (BUN),creatinine (Cr),uric acid (UA) results and provide basis for result concordance in different biochemical systems.Methods The within-run precision and the between-day precision for the median and high values of quality control materials were tested in Beckman LX-20 and Hitachi 7600 chemistry analyzer respectively,and the evaluation criteria were according to the precision requirement of manufacturer and our laboratory.According to American Clinical and Laboratory Standards Institute (CLSI) document EP9-A2,serum BUN,Cr,UA from 40 patients were detected by Hitachi 7600 chemistry analyzer as experimental method and by Beckman LX-20 chemistry analyzer as comparative method.The relative bias ( SE％) between experimental method and comparative method was calculated and the concordance of biochemical analysis system was determined according to EQA requirements and acceptable performance criteria Results The between-run and between-day precision coefficients of variation for BUN,Cr and UA in two biochemical analysis systems were lower than the precision requirement of manufacturer and our laboratory.In Hitachi 7600 chemistry analyzer,the SE％ for BUN at three levels of medical decision were 9.4％,2.0％ and 0.8％,the SE％ for Cr were 7.3％,3.8％ and 2.8％,the SE％ for UA were 7.9％,1.7％ and 1.2％.Two biochemical analysis systems were not comparable in BUN analysis because SE％ of serum BUN levels detected by Hitachi 7600 at low medical decision level exceeded the specified range.The remaining items between two biochemical analysis systems were concordant.Conclusion While the same test is detected by more than two systems in one laboratory,it is necessary to do method comparison and bias evaluation for concordance in order to ensure the comparability among test results.

Objective To evaluate the effects of compound glycyrrhizin on the percentage of Th17 cells and expression of interleukin(IL)-17A in peripheral blood of patients with psoriasis vulgaris, and to explore their relationship with therapeutic effects. Methods A total of 30 patients with mild to moderate progressive psoriasis vulgaris were randomly and equally divided into two groups:group 1 treated with compound glycyrrhizin injection, antihistamines and topical drugs, group 2 treated with antihistamines and topical drugs. Twelve healthy human subjects served as the control group. Blood samples were collected from the patients 1 day before start of treatment and after 3 weeks of treatment, and from the control group. Flow cytometry and enzyme-linked immunosorbent assay (ELISA)were performed to determine the percentage of Th17 cells and expression level of IL-17A respectively in the peripheral blood samples. Results Before the treatment, the percentage of Th17 cells and expression level of IL-17A were both significantly higher in the two patient groups than in the control group (all P<0.05). After the treatment, group 1 showed significant decreases in the percentage of Th17 cells and expression level of IL-17A compared with those before the treatment (both P<0.05), while no significant differences were observed in group 2 between pre-and posttreatment IL-17A expression level or Th17 cell percentage (both P>0.05). Furthermore, both Th17 cell percentage and IL-17A expression were significantly different between the two patient groups after the treatment (both P< 0.05). Conclusion Compound glycyrrhizin may treat psoriasis vulgaris by regulating Th17 cell percentage and IL-17A expression in peripheral blood.

Objective To explore the p27 gene expression and clinical significance in acute leukemia (AL).Methods RT-PCR was used for testing the mRNA levels of p27 gene in bone marrow mononuclear cell of 65 cases of newly diagnosed AL patients and 41 controls.The correlation between the p27 gene expression positive rate and age, sex, peripheral blood white blood cell count and extramedullary infiltration were analyzed simultaneously in AL patients.Results The positive rate of p27 gene expression (36.9 ％,24/65)was significantly lower in AL group than that in the control group (87.8 ％,36/41) (P ＜0.05),and which was lower than the control group (P ＜0.05) in the AML group (34.2 ％,13/38) and the ALL group (37.5 ％,9/24).However,comparing AML group with ALL group,the difference was not statistically significant (P ＞0.05).At the same time,compared the p27 gene expression positive rate of ≥ 14 years old group (52 cases) and ＜14 years old group (13 cases),and also compared in male group (34 cases) and in female group (31 cases),there was no statistically difference (P ＞0.05); Compared the p27 gene expression positive rate of peripheral blood white blood cell count ≥20×109/L group (31 cases) and ＜20×109/L group (34 cases),also compared with in extramedullary infiltration group (37 cases) and in no extramedullary infiltration group (28 cases),the relationship of p27 gene expression positive rate with peripheral blood white blood cell count as well as extramedullary infiltration was a negative correlation (r =-0.284,P ＜ 0.05,r =-0.300,P ＜ 0.05).Conclusion There is p27 mRNA expression deletion in AL patients,and the deletion is present in the AML and ALL at the same time.p27 mRNA expression is negatively correlated with peripheral blood white blood cell count or extramedullary infiltration,which might be one of the mechanisms of occurrence and development of AL.