Objective To analyze effects of tacrolimus on the secretion of chemokines CXCL9 and CXCL10 by γ-interferon (IFN-γ)-simulated HaCaT cells,as well as phosphorylated Janus kinase 1 (p-JAK1) and phosphorylated signal transducer and activator of transcription 1 (p-STAT1),and to explore the mechanism of tacrolimus in the treatment of vitiligo.Methods HaCaT cells were treated with l,10,20,40,60,80,100,120 mg/L tacrolimus solution separately for 4 hours,and methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the cellular proliferative activity.HaCaT cells were divided into 4 groups:blank control group receiving no treatment,IFN-γgroup treated with 500 U/ml IFN-γfor 12 or 48 hours,tacrolimus group treated with 20 mg/L tacrolimus for 4 hours,and tacrolimus + IFN-γgroup treated with 20 mg/L tacrolimus for 4 hours followed by the treatment with 500 U/ml IFN-γfor 12 or 48 hours.Real-time fluorescence-based quantitative PCR was conducted to measure the mRNA expression of CXCL9 and CXCL10,Western blot analysis to determine the protein expression of CXCL9,CXCL10,p-JAK1,and p-STAT1,and enzyme-linked immunosorbent assay (ELISA) to detect the levels of CXCL9 and CXCL10 in the culture supernatants of HaCaT cells.Results Tacrolimus at the maximum concentration of 20 mg/L had no effect on the proliferation of HaCaT cells (P ＞ 0.05).After the pretreatment with 20 mg/L tacrolimus,the mRNA expression of CXCL9 and CXCL10 significantly decreased from 10 369.08 ± 7.99 and 290.02 ± 2.16 to 5 914.33 ± 4.59 and 114.96 ± 0.73,respectively,after the treatment with IFN-γ(both P ＜ 0.01),and the protein expression of CXCL9,CXCL10,p-JAK1,and p-STAT1 also significantly decreased from 8.47 ± 0.29,7.87 ± 0.17,4.20 ± 0.18 and 4.29 ± 0.11 to 7.36 ± 0.13,7.36 ± 0.09,2.60 ± 0.16 and 3.62 ± 0.19,respectively,after the treatment with IFN-γ (all P ＜ 0.01).Moreover,the levels of CXCL9 and CXCL10 in the culture supernatants of HaCaT cells significantly decreased in the IFN-γgroup (1 213.36 ± 0.95,1 722.41 ± 2.57,respectively) compared with the tacrolimus + IFN-γ group (426.45 ± 0.31,554.12 ± 0.56,respectively,both P ＜ 0.01).Conclusion Tacrolimus can inhibit the secretion of CXCL9,CXCL10,p-JAK1 and p-STAT1 by HaCaT cells stimulated by IFN-γ.

Objective To analyze primary diseases and risk factors of chronic cough in children,and develop clinical thinking for the doctor,looking for the orderly diagnosis method.Methods The clinical data of 123 children with chronic cough(medical history,physical examination,routine chest X-ray,PPD test,mycoplasma,Chlamydia antibody,antibody of respiratory syncytial virus,adenovirus IgM determination of IgM determination,Coxsackie virus IgM determination,when necessary,be lung CT,CT of paranasal sinuses,gastrointestinal barium meal,bronchiectasis agent diagnostic treatment and surgery consultation) were retrospectively analyzed.The cause of chronic cough in children with primary disease and related factors were analyzed.Results 123 cases of chronic cough in children's primary diseases were asthma-related cough in 57 cases (46.3 %),upper airway cough syndrome (rhinitis,allergic rhinitis,sinusitis) in 41 cases (33.3 %),chronic pharyngitis and tonsillitis,bronchitis in 27 cases (22.0%) ; the main etiology for Mycoplasma Chlamydia(48.8%,60 cases adenovirus),19 cases(15.4%),12 cases of respiratory syncytial virus (9.8%).Conclusion The main primary disease cough,asthma associated upper airway cough syndrome,chronic pharyngitis and tonsillitis,bronchitis,chronic cough in children,the main pathogen Chlamydia,Mycoplasma-for adenovirus,respiratory syncytial virus infection,diagnosis should be based on detailed,comprehensive medical history and physical examination,from simple to complex,according to from low to high,from conventional to special,from noninvasive to invasive principles are examined.