Objective To study the association between human papillomavirus (HPV) genotypes and viral load and clinical features of verruca vulgaris.Methods Tissue samples were collected from 48 outpatients with verruca vulgaris,and DNA was extracted from these tissue samples.To determine the genotype of HPV,PCR was performed to amplify the L1 fragment of HPV with universal primers followed by bidirectional sequencing and BLAST.The genotyping results were validated by PCR with type-specific primers.Real-time fluorescence-based quantitative PCR was conducted to measure the viral load of HPV,and hematoxylin and eosin (HE) staining to observe histological changes in these tissue specimens.Results The L1 fragment of HPV was amplified from 35 out of the 48 tissue specimens.Of the 35 L1-positive specimens,32 harbored HPV 7,1 harbored HPV 57,and 2 harbored both HPV 2 and HPV 7.Multiple lesions were observed on extremities in the patient infected with HPV 57,but on the head,face and trunk in the patients coinfected with HPV 2 and HPV 7.There were no significant differences in HPV viral load or vacuolated cell number between patients with single lesions and those with multiple lesions,or between patients with a clinical course of ＜ 6 months and those with a clinical course of 6-12 months.However,HPV viral load tended to decrease one year after the onset,and there was pronounced hyperkeratosis and less vacuolated cells in lesions of long duration (more than 2 years) compared with those of short duration (less than 2 years).Conclusions HPV 7 appears to be the most common HPV genotype associated with verruca vulgaris,and HPV 7 infection usually occurs on the head and face.For verruca vulgaris of less than 1 year,neither HPV viral load nor vacuolated cell number is associated with the count or clinical course of warts.

Objective To investigate the therapeutic efficacy and safety of carbon dioxide (CO2) fractional laser in treating periorbital syringoma.Methods Twenty-eight patients with syringoma were treated with ultrapulse CO2 fractional laser.Digital photos were taken before the treatment,1 month and 3 months after the treatment to evaluate the efficacy.The recovery process and side effects were also recorded to assess the safety.Dermatologists rated the efficacy in 0-4 points scales.The patients assessed overall satisfaction in 0-4 point scales.Results One month after the treatment,74.99 ％ of the patients had improvement rate of over 50 ％ and 67.85 ％ of the patients were satisfied or very satisfied with the overall efficacy.Three months after treatment,78.57 ％ of the patients had improvement rate of over 50 ％ and 71.42 ％ of the patients were satisfied or very satisfied with the overall efficacy.There was no statistically significant difference between 1 month and 3 months (P＞0.05).Conclusions Carbon dioxide fractional laser is an effective and safe approach to treat syringoma with minimized side effects.

Objective To observe the change of the balance between regulatory T (Treg) cells and T helper 17(Th17) cells in peripheral blood of the patients with serofast syphilis .Methods From January 2010 to September 2012 ,28 patients with serofast syphilis in our department were selected into the observation group and 30 normal persons were selected into control group .Flow cytometry was used to measure the expression of Treg cells ,Th17 cells ,specific transcription factors Foxp3 and ROR-γ in the pe-ripheral blood .The comparative analysis was carried out .Results The proportion of Treg cells in the peripheral blood of the obser-vation group was (34 .02% ± 12 .31% ) ,which was higher than (21 .86% ± 8 .24% ) in the control group(P<0 .05) ,and the pro-portion of Th17 cells in the peripheral blood of the observation group was (3 .08% ± 2 .17% ) ,which was lower than (9 .13% ± 1 .96% ) in the control group(P<0 .01);the expression level of Foxp3 of CD4+ T cells was (2946 .35 ± 995 .63) in the observation group ,which was higher than (2468 .74 ± 869 .63) in the control group(P< 0 .05),and the expression level of ROR-γwas (1438 .29 ± 659 .31) ,which was lower than(1831 .25 ± 426 .37) in the control group(P<0 .05) .All above differences had signifi-cantly statistical significance .Conclusion The Treg/Th17 cell balance is abnormal in peripheral blood of the patients with serofast syphilis .This cellular immunity suppression of human body may be the reason leading to serofast syphilis .

Objective:To investigate the type and severity of skin adverse reactions induced by afatinib in the treatment of lung cancer, and to analyze their correlation with anti-lung cancer efficacy.Methods:A case-case-control study was conducted on lung cancer patients treated with afatinib in ZhongDa Hospital, Southeast University from December 2016 to January 2018. The type and severity of skin adverse reactions were evaluated in 76 patients with lung cancer based on the National Cancer Institute-Common Terminology Criteria for Adverse Events (NCI-CTCAE) Version 4.0, and these patients were divided into 3 groups according to the severity of skin lesions, including grade-0, -1, and -2/3 groups. The patients underwent chest computed tomography (CT) examination every 3 months, and the tumor response to afatinib was evaluated according to the Response Evaluation Criteria in Solid Tumors (RECIST) . Anti-lung cancer efficacy of afatinib was compared among the patients with different grades of skin lesions by using the Kruskal-wallis H rank sum test. Results:After treatment with afatinib, 44 of the 76 patients with lung cancer achieved stable condition or partial remission, and 32 experienced disease progression. Skin adverse reactions occurred in 69 patients, and manifested as acneiform lesions in 42 (55.3%) patients, paronychia in 35 (46.1%) , mucosal erosions in 30 (39%) , hair changes in 8 (10.5%) , and hand-foot syndrome in 6 (7.9%) . Improvement was achieved in 3, 7 and 34 cases in the grade-0, -1, and -2/3 groups ( n = 7, 19 and 50 respectively) , respectively. There was a significant difference in the response rate among the 3 groups ( χ2 = 6.117, P = 0.047) , and the response rate was significantly higher in the grade-1 and -2/3 groups than in the grade-0 group (both P < 0.001) , and higher in the grade-2/3 group than in the grade-1 group ( P < 0.001) . Conclusion:The treatment of lung cancer with afatinib can cause various types of skin lesions, such as acneiform lesions, paronychia, mucosal erosions, hair changes and hand-foot syndrome, and the higher the severity of the skin lesions, the more marked the anti-lung cancer efficacy of afatinib.

Objective To establish a model for preparation of tissue-engineered skin grafts with hTERT cells carrying human papillomavirus type 6 (HPV 6) genome in vitro, so as to lay a foundation for studying HPV life cycle. Methods The full-length linear HPV6 genome and plasmid pEGFP-▲EGFP were electrophoretically cotransferred into hTERT cells. After selection using G418 resistance, Southern blotting was performed to determine the viral load of HPV6 in transfected cells. 3T3 J2 trophoblastic cells, type I rat-tail collagen and hTERT cells containing the full-length HPV6 genes (HPV6.hTERT cells)were mixed and cocultured on metal meshes to form skin graft-like structures. Hematoxylin and eosin (HE)staining was performed to observe the structure of formed skin grafts, an immunohistochemical assay to measure the expression of HPV6 L1 protein, and electron microscopy to observe virus particles in the skin grafts. Results The linear HPV6 gene was successfully transferred into hTERT cells, and Southern blotting showed the presence of HPV6 DNA in the transferred hTERT cells. The HPV6.hTERT cells, which were cocultured with 3T3 J2 trophoblastic cells and type I rat-tail collagen, proliferated and differentiated over time, and gradually formed skin grafts giving the appearance of verrucous hyperplasia. HE staining showed that the cocultured HPV6.hTERT cells could form typical stratified structure of skin after 7 days of cultivation, and histopathologic features of HPV infection, including obvious papillomatous hyperplasia, presence of vesicular cells, hyperkeratosis and parakeratosis, could be observed after 21 days. The immunohistochemical assay showed the expression of HPV6 L1 protein in the upper portion of skin grafts, and electron microscopy revealed the presence of HPV6 virus particles in skin grafts. Conclusions The established model for preparation of tissue-engineered skin grafts using HPV 6 genome-carrying cells provides a basis for biological studies of HPV, but its application is limited to some degree.

Objective:To analyze clinical features of coronavirus disease 2019 in patients with autoimmune blistering diseases (AIBDs), and to explore risk factors for the development of severe coronavirus disease 2019 in patients with AIBDs.Methods:Clinical data were collected from patients with AIBDs who were hospitalized in Zhongda Hospital, Southeast University from August 2021 to December 2022, and were retrospectively analyzed. Independent sample t-test, Mann-Whitney U test, or chi-square test (Fisher′s exact test) were used to analyze factors associated with severe coronavirus disease 2019 in AIBD patients. Results:A total of 30 patients with AIBDs accompanied by coronavirus disease 2019 were included, they were aged 62.90 ± 15.72 years, and included 16 males and 14 females; pemphigus vulgaris (12 cases, 40.00%) and bullous pemphigoid (13 cases, 43.33%) predominated in the disease spectrum of AIBDs in these patients. The common clinical manifestations of coronavirus disease 2019 were cough (25 cases, 83.33%), fatigue (22 cases, 73.33%), and fever (15 cases, 50.00%) ; most patients had mild clinical symptoms, while 7 (23.33%) developed into severe coronavirus disease 2019, including 2 out of 14 aged 30 - 59 years and 5 out of 16 elderly patients aged ≥ 60 years, and 2 out of the 5 elderly patients finally died. According to the clinical classification criteria of coronavirus disease 2019, the 30 patients were divided into the mild-to-moderate group (23 cases) and severe group (7 cases) ; the duration of AIBDs was significantly shorter in the severe group (10.71 ± 10.72 months) than in the mild-to-moderate group (27.61 ± 19.67 months, t = 2.16, P = 0.040) ; the remission rate of AIBDs was significantly higher in the mild-to-moderate group (21/23) than in the severe group (4/7, χ2 = 4.36, P = 0.037) ; the average daily dosage of methylprednisolone during the last month was significantly lower in the mild-to-moderate group ( M[ Q1, Q3]: 8 [4, 14] mg) than in the severe group (24 [12, 47] mg, U = 133.50, P = 0.007), and the proportion of patients treated with methylprednisolone at an average dosage of > 8 mg was significantly higher in the severe group (6/7) than in the mild-to-moderate group (8/23, χ2 = 5.59, P = 0.031) . Conclusions:Among the patients with AIBDs accompanied by coronavirus disease 2019, the incidence and mortality of severe coronavirus disease 2019 were higher in elderly patients than in young patients. The short course and no-remission of AIBDs, and the average daily dosage of methylprednisolone being>8 mg/d during the last month were risk factors for the development of severe coronavirus disease 2019 in the patients with AIBDs.

Objective:To measure the skin thickness in patients with erysipelas by high-frequency ultrasonography (HF-USG), and to compare the clinical efficacy of systemic antibiotics alone versus their combination with glucocorticoids in the treatment of erysipelas.Methods:Hospitalized patients with erysipelas were enrolled from Zhongda Hospital, Southeast University from January to December in 2021, and randomly divided into the study group and control group according to the order of visits. The study group was treated with systemic cefathiamidine for 7 days followed by oral methylprednisolone at a dose of 0.4 mg·kg -1·d -1, while the control group was treated with cefathiamidine alone. Before and after the treatment for 10 days, the thicknesses of the epidermis-dermis layers and subcutaneous tissues were measured by HF-USG at the sites of the most severe skin lesions on the affected limbs and at the corresponding sites on the healthy limbs, and white blood cell (WBC) counts, neutrophil (NEU) counts, as well as C-reaction protein (CRP) levels were determined. The t test and non-parametric test were used to compare the efficacy between two groups. Results:A total of 23 patients with erysipelas were enrolled. Among the 12 patients in the study group, 8 were males and 4 were females, and their age was 71.4 ± 11.4 years. Among the 11 patients in the control group, 7 were males and 4 were females, and their age was 67.4 ± 11.1 years. Before treatment, the thicknesses of the epidermis-dermis layers (0.33 ± 0.12 cm) and subcutaneous tissues (1.08 ± 0.49 cm) in the study group were not significantly different from those in the control group (0.25 ± 0.09 cm, 0.98 ± 0.46 cm; t = -1.83, -0.49, P = 0.081, 0.626, respectively). After the 10-day treatment, the thicknesses of the epidermis-dermis layers and subcutaneous tissues of the skin lesions on the affected limbs significantly decreased in both groups compared with those before treatment (both P < 0.05), and the decrease in the thicknesses of subcutaneous tissues was significantly stronger in the study group (0.32 ± 0.33 cm) than in the control group (0.10 ± 0.07 cm; t = 2.20, P = 0.039). Before treatment, the WBC counts ([11.16 ± 4.42] × 10 9/L), NEU counts ([8.26 ± 4.16] × 10 9/L) and CRP levels (median [ Q1, Q3]: 72.20 [19.28, 140.50] mg/L) in the study group were not significantly different from those in the control group ([10.10 ± 4.53] × 10 9/L, [7.21 ± 3.00] × 10 9/L, 34.40 [8.00, 74.20] mg/L, respectively; t or Z = 0.60, 0.71, -0.85, P = 0.578, 0.496, 0.196, respectively). After the 10-day treatment, the WBC counts, NEU counts, and CRP levels significantly decreased in both groups compared with those before treatment (all P < 0.05) . Conclusion:The combined treatment with systemic antibiotics and glucocorticoids could effectively alleviate skin inflammation, and more rapidly reduce the thicknesses of inflamed subcutaneous tissues in patients with erysipelas compared with systemic antibiotics alone.