The generation of transgenic mice by DNA microinjection is a powerful tool to investigate the molecular regulation of gene expression, development, and disease. The power of this technology is that foreign DNA can be introduced into every cell of a developing organism, and the phenotypic impact of this genetic modification can be investigated in a system under the constraints of normal development and physiology. The generation of transgenic mice requires the preparation of the transgene DNA construction, collection of one‐cell fertilized mouse embryos, injection of the transgene into mouse embryos, and transfer of the surviving embryos into a pseudopregnant female. Mice born from such manipulations are then screened for the presence of the transgene. The execution of these procedures requires a highly efficient system; otherwise, the cost of the generation of these mice can be cost prohibitive. However, the production of these animals can provide an invaluable research resource. Curr. Protoc. Mouse Biol. 2:245‐262 © 2012 by John Wiley & Sons, Inc.

A total of 181 citrus-based products, including dried fruits, canned fruits, and fruit juices, collected from China and from abroad in 2021 were analyzed for the four Alternaria toxins (ALTs): alternariol (AOH), alternariol monomethyl ether (AME), tentoxin (TEN), and tenuazonic acid (TeA) via ultra-high-performance liquid chromatography–electrospray ionization–tandem mass spectrometry (UPLC-ESI-MS). Although the concentrations of the four ALTs varied by product and geographically, TeA was the predominant toxin followed by AOH, AME, and TEN. Products made in China showed higher levels of ALTs than those made abroad. Maximum levels of TeA, AOH, and AME in analyzed domestic samples were 4.9-fold, 1.3-fold, and 1.2-fold, respectively, higher than those in imported products. Furthermore, 83.4% (151/181) of the analyzed citrus-based products were contaminated with at least two or more ALTs. There were significant positive correlations between AOH and AME, AME and TeA, and TeA and TEN in all analyzed samples. More importantly, the solid and the condensed liquid products had higher concentrations of ALTs than the semi-solid product samples, as well as tangerines, pummelos, and grapefruits compared to the other kinds of citrus-based products. In conclusion, co-contamination with ALTs in commercially available Chinese citrus-based products was universal. Extensive and systematic surveillance of ALTs in citrus-based products, both domestic and imported, is required to obtain more scientific data for the determination of the maximum allowable concentrations of ALTs in China.

Purpose. To examine the benefits of different numbers of 1064‐nm Nd‐YAG laser treatments in patients with onychomycosis. Methods. This was a pilot study of patients with onychomycosis who were divided into three groups: four treatment sessions (group A), eight sessions (group B), and 12 sessions (group C). Only infected nails of degrees II–III (Scoring Clinical Index for Onychomycosis) were included. Treatment was given once a week using a long‐pulse Nd‐YAG 1064‐nm laser. Patients were followed at 8, 16, and 24 weeks after the first treatment. Side effects were recorded. Results. Treatments were completed for 442 nails in 102 patients. The efficacy rates at 8, 16, and 24 weeks were 35.5%, 38.7%, and 37.4% for group A; 31.4%, 41.7%, and 44.0% for group B; and 27.7%, 50.0%, and 55.4% for group C, respectively. There was a significant difference in the efficacy rate at 24 weeks (P = 0.016) between groups A and C, but not for groups A vs. B, or for groups B vs. C. No difference in the efficacy rate at 8 or 16 weeks was observed among the three groups. In all three groups, the efficacy was better for degree II nails than for degree III nails (all P < 0.05). No side effects occurred. Conclusions. The 1064‐nm Nd‐YAG laser had clinical benefits against onychomycosis. Higher numbers of treatments provided better long‐term (24‐week) benefits, but had no impact on the short‐term outcomes. The efficacy of laser treatment on degree II onychomycosis was better than for degree III.

Objective: To outline the clinical manifestations observed in patients with scabies misdiagnosed as generalized eczema, analyse the factors contributing to these misdiagnoses and explore potential reasons for the resurgence of scabies. Patients and Methods: A retrospective analysis was performed to investigate the patients with scabies misdiagnosed as generalized eczema. Results: We included 23 patients, with twelve (52.17%) being male and eleven (47.83%) female. The illness duration ranged from 0.5 to 7 months. Among all patients, 12 (52.17%) were residents of nursing homes, 5 (21.74%) were staff members of these facilities, 4 (17.39%) were caregivers of long-term hospitalized relatives, 1 (4.35%) was a construction worker, and 1 (4.35%) had a history of tourism. The rash predominantly affected the trunk and extremities, 12 patients (52.17%) are each involved the perineum and fingers webbings. The presentations included erythema, papules, and nodules. The main complaint of all patients was nocturnal itch. Under direct microscopy, 5 patients (21.74%) tested positive for scabies mites, and 3 (13.04%) showed histopathological features consistent with scabies. All patients were initially misdiagnosed with generalized eczema. Conclusion: Over half of all patients diagnosed with scabies either resided or worked in long-term care facilities. The lack of awareness of scabies among medical staff in long-term care facilities readily led to frequent misdiagnosis. Comprehensive measures should be implemented urgently to strengthen disease management.

Vulvar intraepithelial neoplasia (VIN) is a pre-malignant condition of the vulvar skin that is found in 50% to 70% of patients with vulvar squamous cell cancer and is regarded as a precursor of vulvar tumors. Thus far, treatment remains lesion- and patient-specific. Here, we describe a VIN patient who presented with a 15-month history of large lesions in the bilateral labium, associated with human papillomavirus infection. The lesions were inappropriate for surgical excision and laser ablation because of their size; therefore, they were treated with photodynamic therapy and concurrent topical 5-aminolevulinic acid hydrochloride. The patient showed no recurrence throughout 2 years of post-treatment follow-up, and reported only slight pain during treatment. Moreover, no significant side effects or scarring were detected. Thus, we conclude that photodynamic therapy can be a useful alternative treatment for large VIN in the bilateral labium that cannot be excised or ablated.

Trichophyton rubrum is one of the most common types of dermatophyte, causing superficial skin mycosis in human populations. Although laser treatment of onychomycosis has been proven to be effective in the clinic, the underlying mechanism of the effect of the laser on fungal growth is not clear. The objective of the present study was to observe the ultrastructural changes of Trichophyton rubrum following laser irradiation and compare the transcriptome differences between the laser irradiation group and control group. In the present study, scanning electron microscopy and transmission electron microscopy were used to observe the ultrastructural changes following the laser irradiation of Trichophyton rubrum. We also performed RNA‐seq to investigate the effects of laser irradiation on Trichophyton rubrum by comparing the transcriptome pattern with the control. Morphological observation with electron microscopy indicated that laser irradiation resulted in the destruction of the cell membrane system. A significant induction of apoptosis was noted compared with the control group, which was confirmed by the formation of the myeloid body and protein aggregates in the cytoplasm. RNA‐seq demonstrated that the expression levels of Acyl‐CoA N‐acyltransferase and S‐adenosyl‐L‐methionine‐dependent methyltransferase were increased in the laser irradiation group. This result indicated that laser irradiation triggered the initiation of the damage repair pathway. In conclusion, the present study suggested that laser irradiation caused physiological injury and therefore inhibited the growth of Trichophyton rubrum.