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This study aims to develop an improved cell screening system for farnesoid X receptor (FXR) agonists based on a dual luciferase reporter gene system. FXR response element (FXRE) fragments from FXR target genes were cloned and inserted into upstream of firefly luciferase (Luc) gene in the plasmid pGL4-luc2P-Hygro. In combination with the internal reference plasmid containing renilla luciferase, a dual luciferase reporter gene system was developed and used for high throughput screening of FXR agonists. After studying the effects of over-expression of RXR, mouse or human FXR, various FXRE fragments, and different ratio of FXR plasmid amount to reporter gene plasmid, induction efficiency of the screening system was optimized by the known FXR agonist GW4064, and Z factor for the system reached 0.83 under optimized conditions. In summary, an improved cell screening system based on double luciferase reporter gene detection system was developed to facilitate the discovery of FXR agonists, where a new enhanced FXRE element was formed by a superposition of multiple FXRE fragments from FXR target genes, instead of a superposition of traditional IR-1 (inverted repeats-1) fragments.
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Humans , Mice , Animals , Transcription Factors/genetics , DNA-Binding Proteins/genetics , Receptors, Cytoplasmic and Nuclear/genetics , Genes, Reporter , Luciferases/geneticsABSTRACT
Objective:To investigate the body composition and sarcopenia in patients with stable chronic obstructive pulmonary disease (COPD) and to analyze the influencing factors.Methods:A total of 220 patients with stable COPD were enrolled in the study from China-Japan Friendship Hospital during July 2018 to December 2019; 220 age and sex-matched healthy subjects (control group 1) and 220 healthy young adults aged 20-40 years (control group 2) were enrolled from the community. The body composition was measured by bioelectrical impedance method. The demographic characteristics, disease conditions, living background and other related factors were collected by questionnaire. T test or one-way ANOVA were used for comparison between groups, and non parametric test was used for non normal distribution data.Results:Compared with the control group 1 and control group 2, the body fat rate ( Z=-10.037, t=-8.411), the fat free mass index ( Z=-8.165, t=-7.856), and appendicular skeletal muscle index (ASMI) ( t=-7.158, t=-11.989) were significantly lower in stable COPD patients ( P<0.05). Among 220 patients with stable COPD, 51 (23.2%) were diagnosed as sarcopenia; the prevalence of sarcopenia was 18.3% (24/131) in men and 30.3% (27/89) in women (χ2=4.297, P=0.038). The decreased ASMI and grip strength in COPD patients with sarcopenia were significantly associated with age≥75 (χ2= 15.746, F= 14.048), female sex ( Z=5.805, t=2.672), low income ( Z=-4.291, t=-4.789), Global Initiative for Chronic Obstructive Lung Disease (GOLD) grade Ⅳ (χ2=22.644, F=3.905), Modified British Medical Research Council (mMRC) Dyspnea Scale grade 4 (χ2=12.475, F=4.369), not receiving systematic health education ( Z=-4.239, Z=-2.474), no exercise (χ2=14.786, F=3.402), insufficient nutrition intake (χ2=40.531, F=10.529). The range of 6-min walking distance was (110-268) m, that was even shorter for patients with mMRC dyspnea scale grade 4 ( F=3.468, P<0.05). Conclusion:The impairment of body composition is common in COPD patients, which will further affect the body function. It is suggested that the routine evaluation of COPD should include the measurement of body composition.
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Nonalcoholic fatty liver disease (NAFLD) has become one of the most prominent causes of chronic liver diseases and malignancies. However, few therapy has been approved. Radix Bupleuri (RB) is the most frequently used herbal medicine for the treatment of liver diseases. In the current study, we aim to systemically evaluate the therapeutic effects of saikosaponin A (SSa) and saikosaponin D (SSd), the major bioactive monomers in RB, against NAFLD and to investigate the underlying mechanisms. Our results demonstrated that both SSa and SSd improved diet-induced NAFLD. Integrative lipidomic and transcriptomic analysis revealed that SSa and SSd modulated glycerolipid metabolism by regulating related genes, like
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Objective To evaluate the efficacy and safety of bronchoalveolar lavage( BAL) in the treatment of neonatal atelectasis with fiberoptic bronchoscopy under ultrasound monitoring. Methods From June 2018 to December 2018,29 children were diagnosed as atelectasis by lung ultrasound. After conventional mechanical vibration and sputum ineffective,BAL was treated with fiberoptic bronchoscopy. All patients be-fore operation were monitored by ultrasound to find the lung segment where the atelectasis was located. 0. 9%NaCl solution was injected by fibrobronchoscope(1～2 ml/kg),and then sucked to ensure the recovery rate of the lavage fluid was more than 50%. After each lavage,ultrasound was immediately used to monitor the recovery of atelectasis to determine whether or not to continue the lavage. One course of treatment could be continuously performed BAL 1 to 3 times a day,1 course per day,and up to 3 courses of lavage. We analyzed the efficacy,adverse reactions and complications of BAL in the treatment of neonatal atelectasis under ultra-sound monitoring. Results Twenty-nine patients underwent BAL treatment with fiberoptic bronchoscopy,25 cases (86. 2%) were cured,3 cases (10. 3%) were effective,and 1 case (3. 4%) was ineffective. All chil-dren had stable vital signs during treatment. Among them,11 cases (37. 9%) had transient hypoxemia, 3 ca-ses (10. 3%) had tracheal mucosal injury, and 2 cases (6. 9%) had hoarseness. There were no serious com-plications such as pulmonary hemorrhage,pneumothorax,and cardiac arrest. Conclusion BAL treatment of atelectasis under lung ultrasound monitoring has obvious effect,easy to operate,no radiation,no obvious ad-verse reactions and complications,which is worthy of clinical application.
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Objective To observe the effect of acupoint application of Chinese medicine on preventing constipation after stroke. Methods By means of randomized, double-blind, controlled study design, 96 patients of yang-syndrome stroke were randomly divided into observation group and control group, 48 cases in each group. Both groups were given conventional treatment for stroke and routine nursing for preventing constipation, and additionally, the control group was given acupoint application of placebo while the observation group was given acupoint application of self-made umbilical paste of Radix et Rhizoma Rhei(Dahuang) and Natrii Sulfas (Mangxiao). The incidence of constipation, frequency of laxative usage and adverse reactions of the two groups were observed. Results (1) The incidence of constipation of the observation group was lower than the control group on treatment day 7 and 14 , the difference being statistical significant(P 0.05). The observation group had better effect on improving stool properties(P 0.05). The adverse reaction of both groups was mild, and had no effect on the accomplishment of the treatment. Conclusion Acupoint application of Chinese medicine is effective and safe on preventing constipation after stroke, showing good prospects for clinical application.
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Objective To investigate the characteristics of cisplatin resistance in human gastric cancer cell line SGC-7901 . Methods Single cell gel electrophoresis ( SCGE ) was used to measure the level of DNA damage and repair in gastric cancer cell line SGC-7901 and gastric cancer cisplatin resistance cell line SGC-7901/DDP by ob-serving the tail length. Morphological changes of SGC-7901 and SGC-7901/DDP were recorded to evalutate the differences between the two lines. The degree of migration of SGC-7901/DDP and SGC-7901 measured by cell wound scratch assay was used to estimate the ability of invasion. MTT assay was performed to determine the drug sensitivity, IC50 values and cross-resistance of SGC-7901/DDP treated with 5-FU, VP-16, ADM, TAX and LOHP separately. Results The level of DNA damage and repair in SGC-7901/DDP was higher than that in SGC-7901 ac-cording to the tail length of SCGE tests ,suggesting the relationship between cisplatin resistance and the abiity of DNA damage and repair. There was a much smaller volume in SGC-7901/DDP compared with SGC-7901,and clone aggregation always appeared in SGC-7901/DDP. Cell wound scratch assay showed that the migration of SGC-7901/DDP was weaker than that of SGC-7901. MTT showed the significant increase of IC50 and cross resitance in SGC-7901/DDP compared with SGC-7901 treated with 5-FU, VP-16, ADM, TAX and LOHP simultaneously. Conclu-sion The ability of DNA damage and repair in gastric cancer cisplatin resistance cell line SGC-7901 is enhanced significantly. The SGC-7901/DDP shows a notable promotion on multidrug resistance in vivo, and the migration of SGC-7901/DDP is weaker than that of SGC-7901 .
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Objective:To observe the clinical effect of acupuncture therapy to support yang and consolidate the constitution for cervical spondylosis of vertebral artery type (CSA). Methods:A total of 72 patients with CSA were randomly divided upon the random digital table into an acupuncture- moxibustion group and an acupuncture group, 36 cases in each group. The acupuncture-moxibustion group was treated by electroacupuncture (EA) at bilateral Fengchi (GB 20) and C4-6 Jiaji (EX-B 2), in combination of moxibustion at Baihui (GV 20), Shenshu (BL 23) and Mingmen (GV 4) to support yang and consolidate constitution. The acupuncture group was given by routine EA at bilateral Fengchi (GB 20), cervical Jiaji (EX-B 2), Tianzhu (BL 10), Jianjing (GB 21), Houxi (SI 3), Hegu (LI 4) and Waiguan (TE 5). The treatment was given once every day and continuous six sessions made one course. After 2-course treatment, the improved situation of clinical symptoms and clinical effect were observed in the two groups. Results:The total effective rate was 91.7% in the acupuncture-moxibustion group and 61.1% in the acupuncture group. The difference of the total effective rate between the two groups was statistically significant (P<0.05). After treatment, the scores of the clinical symptoms in the two groups were obviously decreased than those of the same group before treatment (P<0.01). The differences in the scores between the two groups were statistically significant (P<0.05). Conclusion:Acupuncture-moxibustion treatment to support yang and consolidate constitution is better than the routine acupuncture therapy in the therapeutic effect for CSA.