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.

Humans ; Inservice Training ; Occupational Health ; Safety

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

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

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.

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

·Age-related macular degeneration (AMD) and retinitis pigmentosa (RP) are common outer retinal degenerative problems, which are also the predominant causes of most blinding retinal diseases. Retinal prosthesis is a promising solution for such photoreceptor degeneration diseases.Most of current concepts for a retinal prosthesis are based on neuronal electrical stimulation. In the past twenty years, retinal prosthesis has been developed in two different directions: epiretinal prosthesis and subretinal prosthesis. Each prosthesis technique has its advantages and disadvantages. For epiretinal prosthesis, it is easier to be implanted and has the advantage of keeping most of the electronics in the vitreous cavity, off the retinal surface, which greatly helps in dissipating the heat generated by the implant device. In this paper, a brief overview of retinal prostheses concepts is introduced. After that, several important aspects of epiretinal electrical stimulation will be discussed. Moreover, some practical epiretinal prosthesis devices developed by researchers in United States, Germany and Japan in the past have been reviewed. We hope that the devices will be used widely in the near future.

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

BACKGROUND:Studies have reported that the superfamily of transforming growth factors exert a role in the mineralization of various stem cells, but the combination effects of transforming growth factorβ3 and heparin on proliferation and mineralization ability of stem cells from human exfoliated deciduous teeth remains to be studied. OBJECTIVE:To explore the effect of transforming growth factorβ3 on odontoblast-like differentiation of stem cells from human exfoliated deciduous teeth. METHODS:Human deciduous teeth were col ected using enzyme digestion. The 3rd cells were incubated with 25μg/L recombinant human transforming growth factorβ3, 10 U/mL heparin or their combination. The dentin sialophosphoprotein mRNA and dentinsialoprotein expressions were detected by Q-PCR and western blot assay. Alkaline phosphatase activity was determined using alkaline phosphatase kit. RESULTS AND CONCLUSION:Stem cells from human exfoliated deciduous teeth grew wel after induction. The activity of alkaline phosphatase in the combination group was significantly higher than that in the transforming growth factorβ3, heparin and control groups (P<0.01). After combination induction, the cells were strongly positive for alizarin red staining. Results fromα-PCR and western blot assay showed that the expressions of dentin sialophosphoprotein were both remarkably increased at mRNA and protein levels. In summary, stem cells from human exfoliated deciduous teeth can differentiate into odontoblast-like cells under the induction of transforming growth factorβ3 plus heparin.

BACKGROUND:The role of transforming growth factorβsuperfamily has been reported in bone mineralization of various types of stem cells, but the effects of transforming growth factorβ3 (TGF-β3) combined with heparin on proliferation and mineralization of dental pulp stem cells from human deciduous teeth remains to be studied. OBJECTIVE:To evaluate the effects of TGF-β3 on the proliferation and mineralization of dental pulp stem cells from human deciduous teeth. METHODS:The enzyme digestion method was utilized to separately culture dental pulp stem cells from human deciduous teeth. The cellcolony forming efficiency was determined. Flow cytometry was utilized to identify cellsurface marker CD146. Immunochemistry for Vimentin and STRO1 was performed to measure dental pulp stem cells from human deciduous teeth. The third passage dental pulp stem cells from human deciduous teeth cultured in vitro were intervened with heparin and TGF-β3 of 1, 5, 25μg/L mass concentration. The MTS method was applied to measure cellgrowth curves. Alizarin red staining was carried out. The changes in alkaline phosphatase activity were determined with alkaline phosphatase kit. RESULTS AND CONCLUSION:The cellcolony forming efficiency was high. cells were positive for CD146, and strongly positive for Vimentin and STRO1. Dental pulp stem cells from human deciduous teeth were identified. MTS assay indicated that there was no obvious effect on promoting proliferation of dental pulp stem cells from human deciduous teeth after stimulation of TGF-β3. Detection results of alkaline phosphatase activity demonstrated that the combination of TGF-β3 and heparin could strengthen the alkaline phosphatase activity of dental pulp stem cells from human deciduous teeth with increased concentration. Alkaline phosphatase activities were significantly higher in the TGF-β3+heparin group, TGF-β3 group and heparin group than in the control group (P<0.01). Alizarin red staining was positive in the TGF-β3+heparin group, and the staining was strongest in the 5μg/L TGF-β3+heparin group. Results indicated that TGF-β3 combined with heparin promoted mineralization of dental pulp stem cells from human deciduous teeth.