Objective This study analyzed the chemical structures of two unknown substances in e-liquid using methods such as nuclear magnetic resonance(NMR),infrared spectroscopy(IR),and mass spectrometry(MS).Methods After separating,purifying,and freeze-drying two suspicious substances from e-liquid,GC-MS analysis was used to obtain the mass spectrum characteristic ion fragments and fragmentation patterns of unknown compounds.The precise mass numbers were measured by high-resolution mass spectrometry,and proton numbers and their assignments,carbon types,and fluorine atom numbers and positions were obtained by combining 1HNMR,13CNMR,and 19FNMR.The functional groups were then analyzed by infrared spectroscopy.Results Through data analysis,the molecular formulas of the two suspicious substances are C13H13FN2O2 and C14H15FN2O2,respectively.Their structures were determined to be etomidate derivatives:1-(o-fluoromethylphenyl)imidazole-5-carboxylic acid methyl ester and 1-(o-fluoromethylphenyl)imidazole-5-carboxylic acid ethyl ester.Conclusion The structural data of two substances obtained in this study in terms of mass spectrometry,nuclear magnetic resonance,and infrared spectroscopy provide support for combating crimes involving the two substances,and provide reference for the regulation and detection of the two drugs.

Objective This study analyzed the chemical structures of two unknown substances in e-liquid using methods such as nuclear magnetic resonance(NMR),infrared spectroscopy(IR),and mass spectrometry(MS).Methods After separating,purifying,and freeze-drying two suspicious substances from e-liquid,GC-MS analysis was used to obtain the mass spectrum characteristic ion fragments and fragmentation patterns of unknown compounds.The precise mass numbers were measured by high-resolution mass spectrometry,and proton numbers and their assignments,carbon types,and fluorine atom numbers and positions were obtained by combining 1HNMR,13CNMR,and 19FNMR.The functional groups were then analyzed by infrared spectroscopy.Results Through data analysis,the molecular formulas of the two suspicious substances are C13H13FN2O2 and C14H15FN2O2,respectively.Their structures were determined to be etomidate derivatives:1-(o-fluoromethylphenyl)imidazole-5-carboxylic acid methyl ester and 1-(o-fluoromethylphenyl)imidazole-5-carboxylic acid ethyl ester.Conclusion The structural data of two substances obtained in this study in terms of mass spectrometry,nuclear magnetic resonance,and infrared spectroscopy provide support for combating crimes involving the two substances,and provide reference for the regulation and detection of the two drugs.

Objective To detect the expressions of apoptosis-related factors and inflammatory cytokines in superficial and deep layers of as well as anagen hair follicles in lesions of early alopecia areata (AA).Methods Scalp biopsy samples were collected from 25 patients with early AA and 15 healthy human controls.Fluorescence-based quantitative PCR was performed to detect mRNA expressions of apoptosis-related genes p53,caspase 3,Fas,survivin and bcl-2,as well as those of inflammatory cytokines interleukin (IL)-4,IL-10,IL-12 and interferon (IFN)-γ.An immunohistochemical assay was conducted to measure the expression of p53 protein in anagen hair follicles.Results Compared with control skin samples,anagen hair follicles in AA lesions showed significantly increased mRNA expression levels (expressed as 2-△△Ct) of pro-apoptotic factors caspase 3,p53 and Fas (6.78,8.01,9.74,respectively,all P ＜ 0.05),but decreased mRNA expression levels of antiapoptotic factors bcl-2 and survivin (0.08 and 0.03 respectively,both P ＜ 0.01),and similar mRNA expression levels of inflammatory cytokines.There was a significant increase in mRNA expression levels of Th1 cytokines IFN-γ and IL-12 (2.75 vs.1.00,P ＜ 0.05; 85.67 vs.1.00,P ＜ 0.01),but a significant decrease in the expression level of the Th2 cytokine IL-10 (0.002 vs.1.000,P ＜ 0.01) in superficial layers of AA lesions compared with those of normal control skin.The degree of changes in mRNA expression levels of IL-10 and IL-12 was significantly higher in superficial layers than in deep layers of AA lesions (P＜0.01 and 0.05 respectively).The immunohistochemical assay showed that the number of p53-positive cells per 100 cells in anagen hair follicles of AA lesions was higher than that in those of control skin (t =23.79,P ＜ 0.01).Conclusions Anagen hair follicles in AA lesions exhibit high expressions of pro-apoptosis factors,but low expressions of antiapoptotic factors,suggesting that apoptotic factors play a role in the occurrence of AA.

ObjectiveTo assess the relationship of filaggrin expression with atopic diathesis and disease severity in patients with alopecia areata (AA).MethodsThirty-seven patients with AA aged (26.3 ± 10.6) years were enrolled in this study.Atopic diseases were noted in 8 of these patients.Clinical data and laboratory test resuhs were reviewed.Immunohistochemical staining was performed to quantify the expression of filaggrin protein in scalp biopsy specimens from all of the 37 patients with AA and from 10 human controls,and fluorescence-based semiquantitative reverse transcription-PCR to detect the expression of filaggrin mRNA in scalp biopsy specimens from 22 patients with AA and 13 healthy controls.Data were statistically analyzed by Mann Whitney U test,chi-square test,and Spearman's rank correlation test.ResultsThe expressions of filaggrin protein and mRNA were significantly lower in patients with AA than in the controls(P ＜ 0.05 or 0.01 ),and the decrease seemed more obvious in patients with large areas of lesions,long duration of disease,and nail abnormalities,but the degree of decrease was unrelated to the complication with atopic diseases.No significant differences were observed in sex ratio,age at onset,disease duration,area of hair loss,the prevalence of family history or incidence of nail abnormalities and increase in serum IgE and eosinophils,between patients with atopic diseases and those without.ConclusionsThe expressions of filaggrin protein and mRNA are decreased in patients with AA,suggesting that filaggrin may participate in the development of AA and is correlated with the severity of AA.

Objective To observe the microstructural changes in lesions of alopecia areata (AA) with dermoscopy and to evaluate their correlation with clinicopathological manifestations. Methods The area of alopecia of 62 patients with AA and 44 patients with other types of hair loss were observed by using a noncontact polarized dermoscope (Dermlite, USA). Clinical data on and laboratory findings from these patients were collected. Pathological examination was carried out with scalp biopsy specimens from the alopecia area of 15 AA patients. Results Characteristic dermoscopic signs of AA included yellow dots, black dots, broken hairs, exclamation mark hairs, short vellus hair and newly-grown short hairs. Among these signs, yellow dots showed the highest prevalence (83.9%). Exclamation mark hairs, black dots and broken hairs were rather specific signs for AA, and the prevalence of the three signs was positively correlated with disease activity and positivity rate of hair-pull test. A positive correlation was also noted between the prevalence of elevated thyroid peroxidase antibody levels and positivity rate of hair-pull test (r = 0.269, P ＜ 0.05 ) as well as prevalence of broken hairs (r = 0.445, P ＜ 0.05), and between the prevalence of yellow dots and that of keratinous plug in follicular orifice. There was a negative correlation between the prevalence of newly-grown short hairs and perifollicular mast cell infiltration and between the prevalence of black dots and the anagen/catagen ratio. Conclusions Yellow dots can serve as a preliminary screening marker for AA. Exclamation mark hairs, black dots and broken hairs are highly sensitive for the confirmation of diagnosis of AA, and often predict progressive AA.Dermoscopic signs are well correlated to the histopathology features of AA, and may be useful for the evaluation of disease severity and guidance on the treatment of AA.