Objective:To assess the value of culture of epidermal melanocytes from negative-pressure suction blisters in the auxiliary diagnosis of segmental vitiligo-like nevus depigmentosus.Methods:Between June 2019 and March 2020, 8 patients with segmental vitiligo-like nevus depigmentosus, who met the Coupe′s clinical diagnostic criteria, were enrolled from Department of Dermatology, Hangzhou Third People′s Hospital. All patients were evaluated by the Wood′s lamp, reflectance confocal microscopy (RCM) , 308-nm excimer laser radiation, and in vitro culture of epidermal melanocytes from negative-pressure suction blisters. Results:Among the 8 patients, fluorescence was observed in 6 under the Wood′s lamp, dermal papillary rings were incomplete or absent in 4 as shown by RCM, and 5 experienced no repigmentation after 308-nm excimer laser radiation. Among the 8 patients, in vitro cultured lesional melanocytes were all positive for ferrous sulfate staining, yellowish-white precipitates were obtained after digestion and centrifugation of the melanocytes, and stage Ⅰ-Ⅲ melanosomes were observed in the cytoplasm of melanocytes under the electron microscope; however, the precipitates were black in color after digestion and centrifugation of the melanocytes collected from the normal skin tissues at the contralateral anatomical site, and stageⅠ-Ⅳ melanosomes were seen in the cytoplasm of the melanocytes under the electron microscope. Conclusion:Culture of epidermal melanocytes from negative-pressure suction blisters may facilitate the diagnosis of segmental vitiligo-like nevus depigmentosus.

Objective:To investigate the occupational damage to workers exposed to chromate in a steel plant.Methods:In January 2021, a retrospective analysis was used to select 850 workers exposed to chromate (observation group) and 598 workers not exposed to chromate (control group) in a steel plant in Shandong Province from 2016 to 2017 as the investigation. We collected their occupational-related information, blood routine, fasting blood sugar, nasal lesions, skin lesions, chest X-rays and other inspection results, compared the differences in the abnormal detection rate of the two groups of respondents, and analyzed the occupational hazards of chromium workers.Results:Incidence of nasal damage, skin lesion, up-regulation of ALT (Alanine aminotransferase), abnormal chest radiograph, abnormal serum biochemical index, and abnormal serum glucose level were observed higher in the exposed group than those in the control group (χ 2=125.69, 12.25, 5.82, 10.37, 10.46, 20.66, P=0.000, 0.000, 0.016, 0.001, 0.001, 0.000). Among the symptoms, the incidence of erythra, nasal septum deviation, nasal mucosal congestion, nasal mucosal erosion and rhinitis were more frequent than those in the control group (χ 2=101.54, 4.07, 13.20, 32.05, P=0.000, 0.044, 0.000, 0.000). There was no significant increase in the incidence of work type, age, length of work and the area of nasal mucosa erosion in the observation group compared with the control table, and the difference was not statistically significant (χ 2=5.31、0.42、0.28, P=0.505, 0.662, 0.871) . Conclusion:Occupational hazards of long-term exposure to chromate cannot be ignored. Attention should be paid to strengthening occupational protection and health education of workers exposed to chromium, and increasing their attention.

Objective:To investigate the occupational damage to workers exposed to chromate in a steel plant.Methods:In January 2021, a retrospective analysis was used to select 850 workers exposed to chromate (observation group) and 598 workers not exposed to chromate (control group) in a steel plant in Shandong Province from 2016 to 2017 as the investigation. We collected their occupational-related information, blood routine, fasting blood sugar, nasal lesions, skin lesions, chest X-rays and other inspection results, compared the differences in the abnormal detection rate of the two groups of respondents, and analyzed the occupational hazards of chromium workers.Results:Incidence of nasal damage, skin lesion, up-regulation of ALT (Alanine aminotransferase), abnormal chest radiograph, abnormal serum biochemical index, and abnormal serum glucose level were observed higher in the exposed group than those in the control group (χ 2=125.69, 12.25, 5.82, 10.37, 10.46, 20.66, P=0.000, 0.000, 0.016, 0.001, 0.001, 0.000). Among the symptoms, the incidence of erythra, nasal septum deviation, nasal mucosal congestion, nasal mucosal erosion and rhinitis were more frequent than those in the control group (χ 2=101.54, 4.07, 13.20, 32.05, P=0.000, 0.044, 0.000, 0.000). There was no significant increase in the incidence of work type, age, length of work and the area of nasal mucosa erosion in the observation group compared with the control table, and the difference was not statistically significant (χ 2=5.31、0.42、0.28, P=0.505, 0.662, 0.871) . Conclusion:Occupational hazards of long-term exposure to chromate cannot be ignored. Attention should be paid to strengthening occupational protection and health education of workers exposed to chromium, and increasing their attention.

Objective:To investigate clinical efficacy and safety of cultured autologous melanocyte transplantation for the treatment of non-segmental vitiligo accompanied by autoimmune thyroid diseases.Methods:From May 2008 to December 2018, a total of 2 284 patients with non-segmental vitiligo were retrospectively collected, who received cultured autologous melanocyte transplantation in Hangzhou Third People′s Hospital. Among these patients, 75 were also diagnosed with autoimmune thyroid diseases, including hyperthyroidism (42 cases) , hypothyroidism (18 cases) and Hashimoto′s thyroiditis (15 cases) . Efficacy and safety were compared between the vitiligo patients with autoimmune thyroid diseases (concomitant group) and those without (non-concomitant group) . Chi-square test was used to compare enumeration data.Results:Among the 2 284 patients, 1 085 were males and 1 199 were females, with an age of 25.0 ± 1.2 years and a disease duration of 5.1 ± 2.3 years. Six months after transplantation, 1 873 out of 2 209 patients in the non-concomitant group achieved favorable clinical response, with a response rate of 84.8%, including 1 162 achieving complete clinical response (52.6%) ; 46 out of 75 patients in the concomitant group achieved favorable clinical response, with a response rate of 61.3%, including 20 achieving complete clinical response (26.7%) ; the response rate and recovery rate were both significantly lower in the concomitant group than in the non-concomitant group ( χ2 = 29.72, 19.54, respectively, both P < 0.001) . Moreover, the response rate was significantly lower in the hypothyroidism group than in the hyperthyroidism group ( χ2 = 6.61, P = 0.010) . The incidence of isomorphic response at the donor site was significantly higher in the concomitant group than in the non-concomitant group (9.3% vs. 4.3%, χ2 = 4.31, P = 0.038) , so were the recurrence rates of vitiliginous patches at the recipient site after 1, 3, 5 and 10 years (concomitant group: 6.7%, 14.7%, 17.3%, 8.7%, respectively; non-concomitant group: 0.7%, 1.4%, 2.1%, 3.6%, respectively; χ2 = 29.96, 70.69, 67.23, 41.61, respectively, all P < 0.001) . Conclusion:Concomitant autoimmune thyroid diseases negatively affect the efficacy of cultured autologous melanocyte transplantation in the treatment of vitiligo, so effective measures should be taken to prevent isomorphic response and recurrence at the recipient site for patients with non-segmental vitiligo complicated by autoimmune thyroid diseases.

Objective:To study the characteristics of intestinal microflora in patients with vitiligo, and to analyze the relationship between the changes of intestinal microflora and the incidence of vitiligo, so as to provide new ideas for clinical treatment.Methods:Fecal specimens from 30 patients with vitiligo and 30 healthy adults were collected and analyzed qualitatively by Roche/45 high-throughput sequencing platform. At the same time, macrogenomics was used to analyze the feces of 5 patients with vitiligo and 5 healthy adults to identify the potential regulatory pathways.Results:The bacterial species in the feces of patients with vitiligo were similar to those of healthy people, but the intestinal microbial diversity of patients with vitiligo was significantly reduced ( P<0.01); the abundance of Proteus and Clostridium was significantly reduced at phylum level; at genus level, 7 of them were Bacteroides, Escherichia coli Shigella, Rochella, Bacillus anthracis, Clostridium clostridium, Jordani bacteria. The abundance of RF9 and Prunella-7 decreased significantly ( P<0.01), while the abundance of 4 genera (Rumen Coccus-1, Rumen Coccus UCG, Trichomonas and Streptococcus) increased significantly ( P<0.01). The expression of Streptococcus and Phase Anthraceae in vitiligo patients was significantly different: the former increased by 10.8 times, the latter decreased by 6.517 times, and an intestinal microorganism based on 11 vitiligo-related genera was constructed. The random forest model of bacterial flora showed that AUC of the discriminant model was 0.89 in ROC, and macrogenomic analysis showed that the disorders of vitiligo-related bacterial flora were mainly related to immune-related pathways (such as WNT pathway, Notch pathway), energy metabolism, mitochondrial function and amino acid metabolism (such as phenylalanine metabolism). Conclusions:The diversity of bacterial community in intestinal microecological environment of vitiligo patients is significantly different from those in normal people. The imbalance of bacterial community may be involved in the pathogenesis and development of vitiligo. Supplementation of probiotics may be beneficial to the treatment of vitiligo.

Objective:To identify and differentiate cell subsets in the epidermis and dermis of vitiligo skin lesions using single-cell RNA sequencing technology, and to study the relationship between them.Methods:Skin samples were collected from 2 healthy people without immune or systemic diseases and 2 patients with stable non-segmental vitiligo in Department of Dermatology in the Third People′s Hospital of Hangzhou in September 2019. Single-cell transcriptome sequencing was performed on 11 000 cells in all the skin samples by using 10 × Genomics single-cell RNA-Seq technology. Cell subsets were analyzed, screened and counted by using Seurat software.Results:Cluster analysis of gene expression in the 2 normal skin tissues revealed several cell subsets, including keratinocytes, fibroblasts, nerve cells and melanocytes, endothelial cells, tissue stem cells, and immune cells mainly consisting of dendritic cells and T cells. In the 2 vitiligo lesions, abnormal differentiation and quantity were observed in fibroblasts and 4 keratinocyte subpopulations. The proportion of fibroblasts was significantly lower in vitiligo lesions than in normal skin tissues (0 vs. 0.4%) , while the proportions of keratinocyte subpopulations 5, 6, 10 and 12 (8.03%, 7.36%, 3.52%, 0.91%, respectively) in vitiligo lesions were significantly higher than those in the normal skin tissues (4.47%, 3.53%, 2.69%, 0.28%, respectively, all P < 0.01) . Moreover, the above keratinocyte subpopulations were at the end of cell differentiation, and expressed very significant and specific marker genes, which were mainly closely related to cell-cell interactions and cell homeostasis. GO and KEGG analysis showed that keratinocyte subpopulations 5 and 6 were mainly related to intercellular connection, cell adhesion and cytoskeleton function, while the keratinocyte subpopulation 10 was closely related to cell homeostasis. Conclusion:The single-cell sequencing technology was firstly used to study the transcriptional expression profile of vitiligo lesions in China, and preliminary analysis revealed 4 groups of keratinocytes with different quantity and functions, suggesting that abnormal differentiation and dysfunction of keratinocyte subpopulations may affect the occurrence and development of vitiligo.

Objective:To explore a method for detecting recombinant human Annexin A11 (ANXA11) in serum exosomes of pancreatic cancer patients, and then primarily evaluate the clinical value of ANXA11 in pancreatic cancer patients.Methods:A prospective study was conducted and serum specimens from 70 patients diagnosed with PC, 15 patients diagnosed with benign pancreatic mass and 70 patients diagnosed with pancreatitis from the Affiliated Hospital of Nantong University were collected from December 2016 to July 2019. 70 healthy subjects during the same period were selected as control group. The abundance of ANXA11 in serum and exosomes-free serum were detected through parallel reaction monitoring (PRM) basing on high-resolution, high-precision mass spectrometer. Dot immunoblotting created by ourselves for detecting ANXA11 in exosomes and then the methodological evaluation were carried out. Levels of ANXA11 in exosomes in all subjects were statistically analyzed. Moreover, the areas under the curve (AUC) of receiver operating characteristic (ROC) curves were adopted to evaluate the diagnostic efficacy of ANXA11, CA19-9, CEA on PC. The relationship between ANXA11 and clinicopathological parameters as well as prognosis of PC patients was analyzed in the next moment. For analysis, the Mann-Whitney U test was used for comparing between either two groups, and the kruskal-wallis test was used for comparison among four groups. Results:The detection of serum exosome ANXA11 has high sensitivity and repeatability by the method of self-established dot immunoblotting. ANXA11 increased most significantly in the PC group, and the difference was statistically significant ( Hc=58.079, P<0.01) compared with other three groups. ROC curve analysis showed that the diagnostic performance of ANXA11(area under the curve (AUC=0.836) was higher than CEA (AUC=0.656) and equal to CA19-9 (AUC=0.870). The combination of ANXA11 and CA19-9 could improve the sensitivity of diagnosing PC and maintain good specificity. The level of serum exosome ANXA11 before treatment in PC patients was not related to age, gender, tumor size, tumor growth site, lymph node metastasis, distant metastasis and TNM stage ( Z values are 0.052,-0.285,-0.402,0.324,0.888,0.658,1.734, P>0.05). Furthermore, during the 10th day after surgical treatment, the level of ANXA11 showed no statistical difference compared with that before surgery ( Z value is -1.569, P=0.12). The survival time of PC patients was related to the presence of lymph node metastasis, distant metastasis, TNM staging and treatment protocols (χ 2 values are 9.354,6.086,9.389,16.998, P<0.05), while had no correlation with levels of CEA, CA19-9 and ANXA11 (χ 2 values are 1.516, 0.011, 0.159, P>0.05). Conclusions:This study successfully established an original method for detecting ANXA11 levels in serum exosomes of human. Serum exosomes ANXA11 combined with CA19-9 could improve the diagnostic sensitivity of PC.

Objective To evaluate the efficacy of autologous cultured melanocyte transplantation in the treatment of genital vitiligo.Methods A retrospective analysis was carried out.From 2013 to 2016,6 patients with genital vitiligo were enrolled from Department of Dermatology,Hangzhou Third People's Hospital,who received the treatment with autologous cultured melanocyte transplantation.These patients aged 13-29 years,and the age of onset and disease duration ranged from 10 to 19 years and 2 to 10 years respectively.The area of white patches at the genital site ranged from 10 to 25 cm2.Before the transplantation,all the white patches of the 6 patients bad been stable for ＞ 12 months.Two weeks after the treatment,pimecrolimus was applied to the wounds twice a day after crust shedding.When the patient achieved ＞ 90％ repigmentation of the white patches,pimecrolimus was applied once per three days for maintenance treatment.After the treatment,these patients were followed up for at least 24 months,and the longest follow-up period was 60 months.Results Obvious pigmentation was observed in all the 6 patients 2 months after the transplantation.Meanwhile,3 patients achieved 80％ repigmentation,2 achieved 70％ repigmentation,and 1 achieved 50％ repigmentation.Six months after the treatment,3 patients achieved 95％ repigmentation,2 achieved 80％ repigmentation,and 1 still maintained 50％ repigmentation.Twelve months after the treatment,95％ repigmentation was still maintained in the 3 patients,90％ repigmentation was achieved in 2 patients,and 1 patient maintained 50％ repigmentation,and these outcomes lasted until the end of follow-up.During the follow-up,no adverse reactions were observed.Conclusion Autologous cultured melanocyte transplantation is effective for the treatment of stable genital vitiligo.

Objective To analyze the clinical characteristics of connective tissue disease related interstitial lung disease (CTD-ILD).Methods The demographic data,clinical manifestations,blood gas analysis,lung function,chest high resolution CT (HRCT),histopathology of 186 patients with CTD-ILD admitted in Jinhua Central Hospital from January 2013 to December 2017 were retrospectively analyzed.Results In 186 patients with CTD-ILD,70 were males and 116 were females,with a mean age of 56.22± 12.27.Among them,61 cases were polymyositis/dermatomyositis (DM/PM)-ILD,44 cases were rheumatoid arthritis (RA)-ILD,26 cases were interstitial pneumonia with autoimmune features (IPAF),16 cases were systemic sclerosis (SSc)-ILD,14 cases were Sjogren syndrome (SS)-ILD,11 cases were overlap syndrome (OS)-ILD,7 cases were systemic lupus erythematosus (SLE)-ILD,5 cases were undifferentiated connective tissue disease (UCTD)-ILD,and 2 cases were mixed connective tissue disease (MCTD)-ILD.The incidence of respiratory symptoms was higher than that of systemic symptoms,and the incidence of systemic symptoms was higher than that of skin symptoms.Cough was the most common symptom of respiratory system,and arthralgia was the most common symptom of systemic symptoms,accounting for 54.9％(101/186) and 48.9％ (91/186)respectively.Blood gas analysis showed that hypoxemia accounted for 56.5％(83/147) and type Ⅰ respiratory failure accounted for 26.5％(39/147).The proportion of restrictive ventilation dysfunction in pulmonary function tests was 70.9％(105/148) and the diffuse dysfunction was 81.1％(120/148).The manifestations on HRCT were reticular linear shadow (44.1％),ground-glass opacity (54.3％),consolidation (23.1％),nodules shadow (20.9％) and cystic low density shadow(9.1％).The histopathological diagnosis mainly was non-specific interstitial pneumonia (NSIP,43.0％) and usual interstitial pneumonia (UIP,40.0％).Conclusion The arterial blood gas analysis,lung function and chest HRCT screening,and lung histopathological examination are important for early identification and diagnosis of CTD-ILD.

Objective: To investigate the value of enhanced recovery after surgery (ERAS) in D2 radical resection of distal gastric cancer under total laparoscopy and its effect on inflammatory factors. Methods: The clinical data of 50 patients with distal gastric cancer at Hanzhong Central Hospital from March 2016 to October 2017 were retrospectively analyzed. The patients were divided into the observation group (25 cases) and the control group (25 cases). The observation group received totally laparoscopic radical D2 gastrectomy for distal gastric cancer under the guideline of ERAS; the control group received totally laparoscopic radical D2 gastrectomy for distal gastric cancer under the guideline of the traditional method. The changes of perioperative related indexes, postoperative recovery and postoperative inflammatory indexes were compared between the two groups. Results: All 50 patients successfully completed the operation. The operation time was (287.2±52.3) min and (296.8±57.9) min, respectively of the observation group and the control group; the number of lymph node dissection was (34.0±11.0) and (34.6±14.3), respectively of the observation group and the control group. There were no significant differences in the above indexes between the two groups (all P > 0.05). The hospitalization time, the hospitalization cost, the postoperative time of getting out of bed of the observation group was (22.0±2.8) d, (61 044.4±4 692.7) yuan, (12.8±1.8) h. The hospitalization time, the hospitalization cost, the postoperative time of getting out of bed of the control group was (23.9±1.9) d, (63 875.81±4 392.6) yuan, (23.7±8.2) h. And there were statistically significant differences between the two groups (all P < 0.05). The pain score on the 1st day was (5.0±0.8) score and (6.4±1.0) score, and on the 4th day was (2.7±0.9) score and (3.9±0.9) score, respectively of the observation group and the control group, and there was a statistical difference (all P < 0.05). There was no statistical difference in the 7th day pain score between the two groups (P > 0.05). There were no statistically significant differences in the tumor necrosis factor α, interleukin-6 (IL-6), C-reactive protein and procalcitonin on the day before the operation of both groups (all P > 0.05); the levels of above four factors in the observation group were lower than those in the control group on the 1st, 4th and 7th day after the operation, and the differences were statistically significant (all P < 0.05). Conclusion The application of ERAS in totally laparoscopic radical resection of gastric cancer can improve the postoperative condition, significantly reduce the inflammatory reaction of patients, which has a certain value of improving the quality of life of patients.