AIM: To demonstrate the susceptibility of cell apoptosis varies during the progress of cell malig- nant transformation from human being in vitro. METHODS: A SV40T - transfected human bronchial epithelial im- mortalized cell line (called M) was selected in this work, which has acquired some characteristics of malignant trans- formation at the later passage. The alterations of apoptosis and bcl- 2, P53 genes between early and later passage of M cells were investigated by means of TDT labeling in situ, chromosome FISH, RNA and protein testing, etc. RE- SULTS: Incidence of apoptosis induced by cis - platin was significantly lower in later than in early passages of M. Levels of bcl - 2 mRNA and protein in later passages were higher than early passages of M, and overxpression of bcl -2 was accumulated following the development of cellular malignancy. P53 protein level was as high in early as in later passages. CONCLUSION: Overexpression of bcl - 2 decreases the cellular sensitivity to apoptotic inductors plays an important role during progress of carcinogenesis in human bronchial epithelial cancers. The inactivation of P53 protein in the SV40 - T transfected M cell line may be one of reasons of bcl - 2 overexpression, but not associated with the accumulation of bcl - 2 expressed level during cell transformation.

Objective:To investigate the effect of laboratory indicators on hair loss in patients with female pattern hair loss (FPHL).Methods:Patients with FPHL who visited the Outpatient Clinic of the Department of Medical Aesthetics in Hangzhou First People’s Hospital from November 2022 to November 2023 were selected as the study group, and healthy women who matched the age of the study group in the physical examination center during the same period were selected as the control group. The general information of the patient was recorded, and was also tested by trichoscopy to rule out other patterns of alopecia. Representative indicators including testosterone, dehydroepiandrosterone sulfate(DHEA-S), thyroid-stimulating hormone, 25-hydroxyvitamin D, and serum ferritin were selected from laboratory tests for further analysis. Otherwise, the proportion of deficiency in vitamin D(<20 ng/ml) was calculated based on 25-hydroxyvitamin D levels (number of deficiency cases/total number of cases in each group×100%). Count data were presented as samples (percentages), and chi-square test was used for comparison between groups. Normally distributed continuous data were presented with Mean±SD, independent samples t-test was used for comparison between groups, M( Q1, Q3) was used for non-normally distributed continuous data, and Wilcoxon rank-sum test was used for comparison between groups. Multivariate logistic regression was used to analyze the influencing factors of FPHL. P<0.05 was statistically significant. Results:A total of 37 patients were selected in both groups. The mean age was (28.8±1.3) years in the study group and (29.6±0.9) years in the control group ( t=0.49, P=0.625). The body mass index was (22.8±0.4) kg/m 2 in the study group, and (23.5±0.3) kg/m 2 in the control group ( t=1.26, P=0.211). The testosterone level was 0.58 (0.49, 0.79) nmol/L in the study group, and 0.54 (0.50, 0.78) nmol/L in the control group( Z=1.42, P=0.157). The level of DHEA-S was 6.21 (5.18, 9.60) μmol/L in the study group, and 6.20 (5.20, 9.34) μmol/L in the control group ( Z=2.75, P=0.006). The level of thyroid-stimulating hormone was 2.56 (1.55, 3.66) mU/L in the study group and 1.49 (1.05, 2.65) mU/L in the control group ( Z=2.51, P=0.012). The level of 25-hydroxyvitamin D was 15.44 (11.80, 21.20) ng/ml in the study group, and the level of 25-hydroxyvitamin D was 20.32 (12.07, 21.20) ng/ml in the control group ( Z=2.30, P=0.021), and the proportion of 25-hydroxyvitamin D deficiency in the study group was 64.9% (24/37), which was higher than that in the control group [40.5% (15/37)] ( χ2=4.39, P=0.036). The serum ferritin level was 64.44 (39.47, 133.45) μg/L in the study group and 67.75 (52.63, 143.83) μg/L in the control group ( Z=0.70, P=0.484). The results of multivariate logistic regression analysis showed that the risk of FPHL was increased by the high level of DHEA-S and thyroid-stimulating hormone, and the low level of 25-hydroxyvitamin D (all P<0.05). Conclusion:Abnormal level of DHEA-S, thyroid-stimulating hormone, and 25-hydroxyvitamin D may be risk factors for FPHL.

Objective:To investigate the effect of laboratory indicators on hair loss in patients with female pattern hair loss (FPHL).Methods:Patients with FPHL who visited the Outpatient Clinic of the Department of Medical Aesthetics in Hangzhou First People’s Hospital from November 2022 to November 2023 were selected as the study group, and healthy women who matched the age of the study group in the physical examination center during the same period were selected as the control group. The general information of the patient was recorded, and was also tested by trichoscopy to rule out other patterns of alopecia. Representative indicators including testosterone, dehydroepiandrosterone sulfate(DHEA-S), thyroid-stimulating hormone, 25-hydroxyvitamin D, and serum ferritin were selected from laboratory tests for further analysis. Otherwise, the proportion of deficiency in vitamin D(<20 ng/ml) was calculated based on 25-hydroxyvitamin D levels (number of deficiency cases/total number of cases in each group×100%). Count data were presented as samples (percentages), and chi-square test was used for comparison between groups. Normally distributed continuous data were presented with Mean±SD, independent samples t-test was used for comparison between groups, M( Q1, Q3) was used for non-normally distributed continuous data, and Wilcoxon rank-sum test was used for comparison between groups. Multivariate logistic regression was used to analyze the influencing factors of FPHL. P<0.05 was statistically significant. Results:A total of 37 patients were selected in both groups. The mean age was (28.8±1.3) years in the study group and (29.6±0.9) years in the control group ( t=0.49, P=0.625). The body mass index was (22.8±0.4) kg/m 2 in the study group, and (23.5±0.3) kg/m 2 in the control group ( t=1.26, P=0.211). The testosterone level was 0.58 (0.49, 0.79) nmol/L in the study group, and 0.54 (0.50, 0.78) nmol/L in the control group( Z=1.42, P=0.157). The level of DHEA-S was 6.21 (5.18, 9.60) μmol/L in the study group, and 6.20 (5.20, 9.34) μmol/L in the control group ( Z=2.75, P=0.006). The level of thyroid-stimulating hormone was 2.56 (1.55, 3.66) mU/L in the study group and 1.49 (1.05, 2.65) mU/L in the control group ( Z=2.51, P=0.012). The level of 25-hydroxyvitamin D was 15.44 (11.80, 21.20) ng/ml in the study group, and the level of 25-hydroxyvitamin D was 20.32 (12.07, 21.20) ng/ml in the control group ( Z=2.30, P=0.021), and the proportion of 25-hydroxyvitamin D deficiency in the study group was 64.9% (24/37), which was higher than that in the control group [40.5% (15/37)] ( χ2=4.39, P=0.036). The serum ferritin level was 64.44 (39.47, 133.45) μg/L in the study group and 67.75 (52.63, 143.83) μg/L in the control group ( Z=0.70, P=0.484). The results of multivariate logistic regression analysis showed that the risk of FPHL was increased by the high level of DHEA-S and thyroid-stimulating hormone, and the low level of 25-hydroxyvitamin D (all P<0.05). Conclusion:Abnormal level of DHEA-S, thyroid-stimulating hormone, and 25-hydroxyvitamin D may be risk factors for FPHL.

Objective:To investigate the effect and mechanism of type ⅩⅦ collagen (COL17) on hair growth in mice with androgenetic alopecia (AGA).Methods:Forty-eight C57BL/6J mice were used to establish AGA model (the back hair of the mice was removed and dihydrotestosterone solution was applied) and divided into 6 groups of 8 mice each by random number table. Negative control group, injection of saline in the depilated area (single point injection of 0.05 ml, 5 points in total); positive control group, topical application of 5% minoxidil tincture in the depilated area, 1 ml/d; COL17 low, medium and high concentration groups, injection of 0.5, 1.0 and 2.0 mg/ml COL17 in the depilated area respectively (single point injection of 0.05 ml, 5 points in total); type Ⅲ and ⅩⅦ collagen (COL3+ COL17) combined high concentration group, injection of 2.0 mg/ml COL3 and COL17 in the depilated area (single point injection of 0.05 ml, 5 points in total). The total treatment time was 21 days, during which the hair growth of mice in each group was observed and recorded. After 21 days, the skin and subcutaneous tissue in the depilated area of the mice were taken to make pathological sections for HE staining, and the number and morphological changes of hair follicles were observed; fresh skin tissue in the depilated area of the mice was taken for total RNA sequencing analysis, and the differentially co-expressed genes were annotated by gene ontology (GO) functional annotation, Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis and gene set enrichment analysis (GSEA).Results:After 21 days of treatment, compared with the negative control group, the depilation area on the back of the mice in the positive control group, COL17 high concentration group, and COL3+ COL17 combined high concentration group was significantly reduced, and HE staining showed that the number of hair follicles was also significantly increased. Pearson correlation analysis, principal component analysis and cluster heat map between groups showed that COL17 high concentration group had high gene correlation with the positive control group ( R2=0.95, P=0.024), and the gene expression was relatively close, with 3 882 differentially expressed genes (1 705 up-regulated and 2 177 down-regulated) in the two groups, while COL3+ COL17 combined high concentration group had the highest gene correlation with the positive control group ( R2=0.96, P=0.001), and the gene expression was the closest, with 1 289 differentially expressed genes (385 up-regulated and 904 down-regulated). KEGG analysis showed that compared with the negative control group, the positive control group, COL17 high concentration group and COL3+ COL17 combined high concentration group of mice all upregulated Wnt signaling pathway, cell adhesion molecules and hedgehog signaling pathway related to hair growth. GO enrichment analysis suggested that COL17 high concentration group and COL3+ COL17 combined high concentration group had upregulated genes related to skin development and hair cycle. GSEA enrichment analysis found that COL17 high concentration group had upregulated genes related to fibroblast proliferation and interleukin-1 secretion, while COL3+ COL17 combined high concentration group had upregulated genes related to fibroblast migration, clearance of apoptotic cells and accelerated metabolism of reactive oxygen species. Conclusion:Local injection of 2.0 mg/ml COL17 has a certain promoting effect on hair growth in AGA model mice, and the effect is more significant after combined injection of 2.0 mg/ml COL3. Activation of Wnt signaling pathway is one of the main mechanisms of COL17 promoting hair growth.

Objective:To investigate the effect and mechanism of type ⅩⅦ collagen (COL17) on hair growth in mice with androgenetic alopecia (AGA).Methods:Forty-eight C57BL/6J mice were used to establish AGA model (the back hair of the mice was removed and dihydrotestosterone solution was applied) and divided into 6 groups of 8 mice each by random number table. Negative control group, injection of saline in the depilated area (single point injection of 0.05 ml, 5 points in total); positive control group, topical application of 5% minoxidil tincture in the depilated area, 1 ml/d; COL17 low, medium and high concentration groups, injection of 0.5, 1.0 and 2.0 mg/ml COL17 in the depilated area respectively (single point injection of 0.05 ml, 5 points in total); type Ⅲ and ⅩⅦ collagen (COL3+ COL17) combined high concentration group, injection of 2.0 mg/ml COL3 and COL17 in the depilated area (single point injection of 0.05 ml, 5 points in total). The total treatment time was 21 days, during which the hair growth of mice in each group was observed and recorded. After 21 days, the skin and subcutaneous tissue in the depilated area of the mice were taken to make pathological sections for HE staining, and the number and morphological changes of hair follicles were observed; fresh skin tissue in the depilated area of the mice was taken for total RNA sequencing analysis, and the differentially co-expressed genes were annotated by gene ontology (GO) functional annotation, Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis and gene set enrichment analysis (GSEA).Results:After 21 days of treatment, compared with the negative control group, the depilation area on the back of the mice in the positive control group, COL17 high concentration group, and COL3+ COL17 combined high concentration group was significantly reduced, and HE staining showed that the number of hair follicles was also significantly increased. Pearson correlation analysis, principal component analysis and cluster heat map between groups showed that COL17 high concentration group had high gene correlation with the positive control group ( R2=0.95, P=0.024), and the gene expression was relatively close, with 3 882 differentially expressed genes (1 705 up-regulated and 2 177 down-regulated) in the two groups, while COL3+ COL17 combined high concentration group had the highest gene correlation with the positive control group ( R2=0.96, P=0.001), and the gene expression was the closest, with 1 289 differentially expressed genes (385 up-regulated and 904 down-regulated). KEGG analysis showed that compared with the negative control group, the positive control group, COL17 high concentration group and COL3+ COL17 combined high concentration group of mice all upregulated Wnt signaling pathway, cell adhesion molecules and hedgehog signaling pathway related to hair growth. GO enrichment analysis suggested that COL17 high concentration group and COL3+ COL17 combined high concentration group had upregulated genes related to skin development and hair cycle. GSEA enrichment analysis found that COL17 high concentration group had upregulated genes related to fibroblast proliferation and interleukin-1 secretion, while COL3+ COL17 combined high concentration group had upregulated genes related to fibroblast migration, clearance of apoptotic cells and accelerated metabolism of reactive oxygen species. Conclusion:Local injection of 2.0 mg/ml COL17 has a certain promoting effect on hair growth in AGA model mice, and the effect is more significant after combined injection of 2.0 mg/ml COL3. Activation of Wnt signaling pathway is one of the main mechanisms of COL17 promoting hair growth.

Objective:To explore the applicability of the distance between facial marks classification in evaluating the severity of androgenic alopecia in men.Methods:From June to December 2019, the male Chinese with diagnosis of androgenic alopecia were evaluated in the specific clinic of alopecia of Hangzhou First People’s Hospital according to the distance between facial marks and BASP(basic and specific) classification. The classification based on the distance between facial marks measures the distance from the facial marks of the anterior hairline to the horizontal line of the eyebrow and the longest radius of hair loss in the hair rotation center, the hair recession of the patient’s forehead (F), temporal (M) and vertex (V) parts. The hair loss in each region is rated as 0-3 grade from light to heavy, and the final hair loss grading is expressed as FnMnVn, such as F1M2V0. The highest grade of hair loss in F, M and V is the overall grade of hair loss. SPSS 25.0 software was used to statistically analyze the general data of patients, and Kappa test was used to evaluate the consistency between the results of the distance classification and BASP classification. The repeatability of the distance classification was tested by the repetition rate of three hair loss specialists. When two or more specialists gave the same evaluation among the three hair loss specialists, the result was regarded as the standard result. The ease of use of the distance between facial marks classification was tested by the consistency rate between the grading results of two temporary trained general doctors and the standard results.Results:A total of 150 male patients, aged (32.8±7.9) years (19-58 years), were included, of which 99 patients were 24-35 years old, accounting for 66.00%. It can be observed that the onset age was earlier. As assessed in this classification, the patients who participated in the study were graded as mild in 65 cases(43.33%), severe in 58 cases(38.67%), and moderate, which was consistency with the results obtained by BASP classification ( κ=0.573, P<0.001). Three experienced alopecia specialists evaluated 150 patients through the distance between facial marks. The results showed that the repetition rates of frontal, temporal and parietal classification results were 98.00%(147/150), 97.33%(146/150) and 96.00%(144/150), respectively. The repetition rate of the final alopecia classification was 92.00%(138/150), and the repetition rate of the overall alopecia classification was 98.00%(147/150). The consistency rate between the overall alopecia classification results of two temporary trained general doctors and the standard results was 95.92%(141/147) and 96.60%(142/147), respectively, and the consistency rate of the other results was higher than 90.00% except for one general doctor who was 89.86%(124/138) in the final classification. Conclusion:The distance between facial marks classification is a comparatively accurate and easy-to-learn grading method designed for Chinese male androgenic hair loss patients based on objective measurement data.

Objective:To explore the applicability of the distance between facial marks classification in evaluating the severity of androgenic alopecia in men.Methods:From June to December 2019, the male Chinese with diagnosis of androgenic alopecia were evaluated in the specific clinic of alopecia of Hangzhou First People’s Hospital according to the distance between facial marks and BASP(basic and specific) classification. The classification based on the distance between facial marks measures the distance from the facial marks of the anterior hairline to the horizontal line of the eyebrow and the longest radius of hair loss in the hair rotation center, the hair recession of the patient’s forehead (F), temporal (M) and vertex (V) parts. The hair loss in each region is rated as 0-3 grade from light to heavy, and the final hair loss grading is expressed as FnMnVn, such as F1M2V0. The highest grade of hair loss in F, M and V is the overall grade of hair loss. SPSS 25.0 software was used to statistically analyze the general data of patients, and Kappa test was used to evaluate the consistency between the results of the distance classification and BASP classification. The repeatability of the distance classification was tested by the repetition rate of three hair loss specialists. When two or more specialists gave the same evaluation among the three hair loss specialists, the result was regarded as the standard result. The ease of use of the distance between facial marks classification was tested by the consistency rate between the grading results of two temporary trained general doctors and the standard results.Results:A total of 150 male patients, aged (32.8±7.9) years (19-58 years), were included, of which 99 patients were 24-35 years old, accounting for 66.00%. It can be observed that the onset age was earlier. As assessed in this classification, the patients who participated in the study were graded as mild in 65 cases(43.33%), severe in 58 cases(38.67%), and moderate, which was consistency with the results obtained by BASP classification ( κ=0.573, P<0.001). Three experienced alopecia specialists evaluated 150 patients through the distance between facial marks. The results showed that the repetition rates of frontal, temporal and parietal classification results were 98.00%(147/150), 97.33%(146/150) and 96.00%(144/150), respectively. The repetition rate of the final alopecia classification was 92.00%(138/150), and the repetition rate of the overall alopecia classification was 98.00%(147/150). The consistency rate between the overall alopecia classification results of two temporary trained general doctors and the standard results was 95.92%(141/147) and 96.60%(142/147), respectively, and the consistency rate of the other results was higher than 90.00% except for one general doctor who was 89.86%(124/138) in the final classification. Conclusion:The distance between facial marks classification is a comparatively accurate and easy-to-learn grading method designed for Chinese male androgenic hair loss patients based on objective measurement data.