ObjectiveTo compare the effectiveness of ω-3 fatty acids and middle/light fatty acids in the intervention of rats model of bleomycin-induced pulmonary fibrosis.MethodsTotally 120 rats were randomly divided into 4 groups: normal saline (NS) group,bleomycin-induced pulmonary fibrosis without treatment group ( BLM group),middle/light fatty acids group,ω-3 fatty acids group.Lung tissues were obtained on the 7th,14th,and 21st day after modeling.The left lung were measured by using immunohistochemical methods for transforming growth factor β1 (TGF-β1) and interferon garmma ( IFN-γ).The lower lobe of the right lung underwent HE staining.Serum TGF-β1,IFN-γ,and interleukin-4 (IL-4) were measured using double-antibody sandwich ELISA.ResultsThe pulmonary alveolitis and fibrosis in the ω-3 fatty acids group was significantly milder than in middle/light fatty acids group and BLM group.On the 7th,14th,and 21st day after modeling,stronger TGF-β1 protein expression was detected in the bronehiolar epithelia of middle/light fatty acids group and BLM group and poorer IFN-γexpression in both groups.However,the opposite results were found in the ω-3 fatty acids group: on the 7th,14th,and 21st day after modeling,TGF-β1 protein expression ( 13.60 ± 5.90,10.53 ± 4.21,and 7.23 ± 2.21 )was lower ( P =0.047) and IFN-γ ( 13.85 ± 7.48,15.32 ± 2.12,and 18.74 ± 2.65 ) was stronger in ω-3 fatty acids group (P =0.041).On the 7 ,14,and 21st day after modeling,the serum IL-4 levels in the middle/light fatty acids group and BLM group became significantly higher,while the IFN-γ level in both groups was significantly lower than in ω-3 fatty acids group ( P =0.008 ) ; meanwhile,in the ω-3 fatty acids group,the serum IL-4 levels [ (8.73 ± 1.20),(5.73 ±2.03),and (4.98 ± 1.89) pg/ml] were significantly lower (P =0.044) and serum IFN-γlevels [ (5.67 ± 0.13 ),( 6.58 ± 0.64 ),and ( 7.05 ± 0.52 ) pg/ml ] were significantly higher ( P =0.048 ) on the 72,14th,and 21st day after modeling.Conclusionsω-3 fatty acids can lower TGF-β1 protein expression in rat lung tissue and reduce the surum TGF-β1 and IL-4 levels.Compared with the middle/light fatty acids,it can more effectively upregulate the expression of IFN-γ in lung tissue and increase its serum level,and thus alleviate pulmonary fibrosis in rats.

D5 receptor is a subtype of dopamine D1-like receptor, and it plays a functional role in many neurological disorders. Some natural compounds from Chinese herbs, which were shown to have the property as that of receptor agonist, might provide a rich source in search of new candidates for therapeutic use. For exploring this possibility, we developed a cell-based high throughput (HTS) D5 receptor assay to screen the herb-based natural compound library established in our centre. The D5 receptor plasmid (hD5R/pcDNA3.1) and reporter gene plasmid (4 x CRE/TK/Luci/pGL3) were co-transfected into HEK293 cell line. After G418 being selected, the monoclonal cell lines bearing hD5R and the reporter gene were established and used for agonist screening. To optimize the assay condition, the effects of some factors such as cell number per well, incubation time, and the doses of SKF38393 (a potent selective partial D1-like agonist) were examined by using forskolin, a positive compound for cAMP response element. The best condition for this HTS assay included: the cell number at 5 x 10(4)/mL, the dose of forskolin at 5-20 micromol/L, the dose of SKF38393 at 100 nmol/L-100 micromol/L, and the agonist incubation time at 6 -8 h. Thereafter, water extracts from more than 200 Chinese herbs in our library were screened and three of these water extracts showed positive activity, with higher or similar activity as SKF38393. In conclusion, we have established a cell-based HTS assay for D5 receptor agonist screening, and by use of this HTS assay, 3 Chinese herbs maybe contain components exhibiting D5 receptor agonist property. This work provides an alternative vision of how to use herb medicines and a way to develop potential drugs for treatment of neurological disorders.
Cell Line ; Dopamine Agonists ; isolation & purification ; Drug Evaluation, Preclinical ; methods ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; High-Throughput Screening Assays ; methods ; Humans ; Plants, Medicinal ; chemistry ; Receptors, Dopamine D5 ; agonists

Pre-testing preparation is the basis and starting point of genetic testing.The process includes collection of clinical information,formulation of testing scheme,genetic counseling before testing,and completion of informed consent and testing authorization.To effectively identify genetic diseases in clinics can greatly improve the diagnostic rate of next generation sequencing (NGS),thereby reducing medical cost and improving clinical efficacy.The analysis of NGS results relies,to a large extent,on the understanding of genotype-phenotype correlations,therefore it is particularly important to collect and evaluate clinical phenotypes and describe them in uniform standard terms.Different types of genetic diseases or mutations may require specific testing techniques,which can yield twice the result with half the effort.Pre-testing genetic counseling can help patients and their families to understand the significance of relevant genetic testing,formulate individualized testing strategies,and lay a foundation for follow-up.

With high accuracy and precision,next generation sequencing (NGS) has provided a powerful tool for clinical testing of genetic diseases.To follow a standardized experimental procedure is the prerequisite to obtain stable,reliable,and effective NGS data for the assistance of diagnosis and/or screening of genetic diseases.At a conference of genetic testing industry held in Shanghai,May 2019,physicians engaged in the diagnosis and treatment of genetic diseases,experts engaged in clinical laboratory testing of genetic diseases and experts from third-party genetic testing companies have fully discussed the standardization of NGS procedures for the testing of genetic diseases.Experts from different backgrounds have provided opinions for the operation and implementation of NGS testing procedures including sample collection,reception,preservation,library construction,sequencing and data quality control.Based on the discussion,a consensus on the standardization of the testing procedures in NGS laboratories is developed with the aim to standardize NGS testing and accelerate implementation of NGS in clinical settings across China.

Bioinformatic analysis and variant classification are the key components of high-throughput sequencing-based genetic diagnostic approach.This consensus is part of the effort to develop a standardized process for next generation sequencing (NGS)-based test for germline mutations underlying Mendelian disorders in China.The flow-chart,common software,key parameters of bioinformatics pipeline for data processing,annotation,storage and variant classification are reviewed,which is aimed to help improving and maintaining a high-quality process and obtaining consistent outcomes for NGS-based molecular diagnosis.