3.The effects of bisphosphonate on the peroxisome proliferator-activated receptor γ expression of glucocorticoid-induced osteoporosis
Chinese Journal of Rheumatology 2010;14(11):743-745
Objective To investigate the effects of bisphosphonate on the proliferator-activated receptor γ(PRARγ)expression of glucocorticoid-induced osteoporosis. Methods ① Twenty-two cases of systemic lupus erythematosus(SLE)patients with glucocorticoids(>0.5 mg·kg-1 ·d-1)were divided into 2 groups: 9 cases in the treatment group in which patients received alendronate 70 mg qw., 13 cases in the control group without anti-osteoporosis treatment. All of the patients had bone marrow puncture after 24 weeks, bone tissues were embedded, sliced, and the value of average optical density of PPARγwas tested with immunohistochemistry. ② Human bone mesenchymal stem cells were cultured in the same conditions and stimulated to adipocytes. During the adipogenic process, the cells were divided into four groups, three of which were experimental groups treated with 10-5 mol/L(high-dose group), 10-7 mol/L(medium-dose group)and 10-9 mol/L(low-dose group)bisphosphonate respectively, while the other was the control group without bisphosphonate in the medium. The adipocytes were identified by oil Red O stain seven days later. The adipocyte rate was measured by light absorption at 515 nm. The expression of PPARγ was measured by quantitative PCR. Comparisons between groups were tested by One-Way ANOVA analysis and t test. Results The average optical density of PPAR-gamma immunohistochemistry in the treatment group was less than the control group(P<0.05). Seven days later, the absorbance between adipogenic induction group and low-dose group was not significant(P=0.167). but the adipogenic absorbance between the high-dose group, middle dose group and the control groups(P values were 0, 0.041). There was significant difference in the quantitative PCR results between the high-dose group, middle dose group and the expression of PPARγ adipogenic induction group(P value was 0, 0.01), but there was no significant differences between the low-dose group and adipogenic induction group(P>0.05). Conclusion Effective concentrations of bisphosphonate can inhibit the expression of PPARγ which in turn lead to the suppression of hMSCs to the adipocytes. The glucocorticoid-induced osteoporosis is also depressed.
4.Expression of hypoxia-inducible factor-1 α in children with attention deficit hyperactivity disorder and its relationship with respiratory function
Chinese Journal of Primary Medicine and Pharmacy 2017;24(9):1390-1393
Objective To study the expression of hypoxia-inducible factor-1 α (HIF-1 α) in children with attention deficit hyperactivity disorder(ADHD),and its relationship with respiratory function.Methods Sixty children with ADHD confirmed by DSM-IV were admitted as ADHD group.60 healthy children were selected as the normal control group.The serum level of HIF-1α was determined by enzyme-linked immunosorbent assay(ELISA).Respiratory rate and respiratory amplitude were measured by biofeedback instrument.The PEF was measured by peak speed meter.The differences of HIF-1 α level and respiratory function between different ADHD groups and control group were analyzed.The influence of age and sex on HIF-1 α level and respiratory function was analyzed.Results The serum levels of HIF-1 α in ADHD group and healthy control group were (43.22-± 11.68) ng/L and (21.48 ± 12.70)ng/L respectively,the respiration rates were (17.47 ±-2.82)/min and (15.28 ± 2.35)/min respectively.The HIF-1 α and respiration rate of ADHD group were higher than healthy control group (t =9.003,4.363,all P < 0.05).The respiratory amplitude in ADHD group and healthy control group were (2.36 ± 1.18) cm and (3.03 ± 1.05)cm respectively,the PEF in the two groups were (191.42 ± 30.06) L/min and (206.62 ± 33.35) L/min respectively.The respiratory amplitude and PEF of ADHD group were lower than healthy control group (t =4.007,8.534,all P <0.05).There were no significant differences in HIF-1α level,respiratory rate,respiratory amplitude and PEF between boys and girls with ADHD (P > 0.05).There were no significant differences in HIF-1 α level,respiratory rate,respiration amplitude and PEF between different age in ADHD children (P > 0.05).There was a negative correlation between HIF-1 α and PEF in children with ADHD (r =0.422,P < 0.05).Conclusion HIF-1 α and lung function are correlated with ADHD,ADHD children have small lung capacity,which may result in the hypoxia of the brain and the increase of HIF-1 α expression.
5. Preparation of recombinant firefly luciferase by a simple and rapid expression and purification method and its application in bacterial detection
Academic Journal of Xi'an Jiaotong University 2010;22(2):97-101
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 1.995 mg/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 (4 500 CFU/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.
6. Preparation of recombinant firefly luciferase by a simple and rapid expression and purification method and its application in bacterial detection
Academic Journal of Xi'an Jiaotong University ;22(2):97-101
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 1.995 mg/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 (4 500 CFU/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.
7.THE APPLICATION OF IMPROVED STANDARDIZED EXECUTION IN PREVENTION AND CONTROL OF HOSPITAL INFECTION
Modern Hospital 2015;(3):113-114
Objective To study and summarizes the methods ofthe standardized hospital infection manage -ment technology in order to prevent and control hospital infection scientifically .Methods Taking the chance of im-provingthe execution of the standardized hospital infection management issued by the Ministry of Health ,we realized organizational execution by enhancing the responsibility decomposition , mechanism implementation by standardized completion,ideological implementation by education and training ,the implementation of standards by inspection and monitoring and finally form a team by enhanced communication .Results The technicalstandards of hospital infection management was implemented and the quality of hospital infection control was improved progressively .Conclusion It is an effective way to guarantee medical security to improve the execution of the technical standard of hospital infec -tion management .
8.Research advance in retinoblastoma
Chinese Journal of Experimental Ophthalmology 2012;30(3):278-282
Retinoblastoma(RB) is the most common infant malignant tumor of eye.It seriously affects the life quality and life span.The rapid development of biological technology allows some new breakthroughs in the basic and clinical researches of RB.Current researches focus on its etiology and pathogenesis,and the ultimate aim is to guide clinical prevention,detection and management.Some literature revealed and summarized the histological source of RB,mutation detection of Rb gene,hereditary clue of RB susceptibility,and gene-therapy of RB.Gene therapy of RB is a new treating approach to RB in recent years,and its development brings a chance for achieving the goal of curing RB and improving patient prognosis.The research progress of RB is reviewed here.
10. Effects of tissue factor pathway inhibitor-2 on the growth and angiogenesis of subcutaneous tumor xenografts in nude mice established from human liver cancer cells
Tumor 2013;33(2):157-163
Objective: To investigate the effects of TFPI-2 (tissue factor pathway inhibitor 2) on the growth and angiogenesis of subcutaneous tumor xenografts in nude mice established from human liver cancer cells. Methods: The Hep3B cells stably expressing TFPI-2 (Hep3B-TFPI-2 group) and the Hep3B cells transfected with empty vector PCDNA3.1 (Hep3B-V group) or without transfection (Hep3B-P group) were subcutaneously transplanted into nude mice respectively to generate subcutaneous tumor xenografts. The volume of tumor xenograft was measured every three days, and the growth curve of tumor xenograft was drawn when the subcutaneous tumor xenograft was visible. The nude mice were killed three weeks after transplant, the volume of tumor xenograft was measured, and the total RNAs and proteins in tumor xenografts were extracted. The mRNA and protein expressions of TFPI-2 and VEGF (vascular endothelial growth factor) in tumor xenografts were analyzed by RFQ-PCR (real-time fluorescence quantitative PCR) and Western blotting, respectively. The expression of TFPI-2 protein and the MVD (microvessel density) in tumor xenografts were observed by immunohistochemistry. Results: The eventual tumor volume of tumor xenografts in Hep3B-TFPI-2 group was apparently smaller than those in Hep3B-V group and Hep3B-P group (both P < 0.05). The expression of mRNA and abundance of protein of TFPI-2 in Hep3B-TFPI-2 group were significantly higher than those in the other two groups (P < 0.05); while the expression of mRNA and abundance of protein of VEGF in Hep3B-TFPI-2 group were apparently lower than those in the other two groups. Compared with Hep3B-V group and Hep3B-P group, the inhibitory rates of VEGF protein expression in Hep3B-TFPI-2 group were 19.8% and 23.5%, respectively (P < 0.05). The MVD in Hep3B-TFPI-2 group was apparently lower than those in the other two groups (P < 0.05). Conclusion: TFPI-2 can significantly inhibit the growth and angiogenesis of subcutaneous tumor xenografts in nude mice established from hepatocarcinoma Hep3B cells. Copyright © 2013 by TUMOR.