Objective:To seek the optimal melanin-removal method for hematoxylin and eosin (HE) staining, immunohistochemistry and molecular detection.Methods:Thirty-eight paraffin tissue samples of malignant melanoma diagnosed at the Fujian Cancer Hospital, Fuzhou, China between January 2018 and March 2022 were collected and used to make a tissue microarray. Melanin in these cases was removed using warm hydrogen peroxide, double oxidation depigmentation, modified potassium permanganate-oxalic acid or trichloroisocyanuric acid, followed by HE staining. The cases were divided into two cohorts: one was subject to the one of the above four methods to remove melanin first, followed by immunohistochemistry (SOX-10, Ki-67, HMB45 and Melan A), while the other was subject to immunohistochemical staining first and then a melanin removal. Following that, seventeen melanin-rich paraffin tissue samples were collected and depigmented using the methods described above. DNA extraction was then done, followed by assessments of DNA content and quality. Moreover, the completeness of melanin removal, the effect on HE and immunohistochemical staining, and the quality of DNA were compared between the depigmented methods.Results:Regarding the effectiveness of melanin removal, the modified potassium permanganate-oxalic acid and the warm hydrogen peroxide methods were the most effective, and both showed residual melanin in only 5.26% (2/38) of the cases. The trichloroisocyanuric acid method showed residual melanin in 10.53% (4/38) of the cases. The worst was the double oxidation depigmentation method, which showed pigment residue in 15.79% (6/38) of the cases. For HE staining, the percentage of good staining with the warm hydrogen peroxide method was 92.11%, higher than the other three methods. For immunohistochemical staining, the mean staining scores of immunohistochemistry first followed by melanin removal with modified potassium permanganate-oxalic acid, double oxidation and trichloroisocyanuric acid were 20.84, 26.63 and 35.02, respectively. These immunohistochemical staining scores were higher than those of melanin removal first followed by immunohistochemistry (8.70, 15.41 and 21.22, respectively). The mean staining score of melanin removal by warm hydrogen peroxide method followed by immunohistochemistry was 33.57, superior to that of immunohistochemistry followed by the melanin removal (19.96). Moreover, the staining scores of HMB45, MelanA and Ki-67 with immunohistochemical staining followed by trichloroisocyanuric acid method were 36.45, 33.79, and 36.24, respectively, while the staining score of SOX10 with melanin removal by warm hydrogen peroxide followed by immunohistochemistry was 34.39. The DNA was significantly degraded by modified potassium permanganate-oxalic acid, double oxidation depigmentation and trichloroisocyanuric acid, whereas the mean concentration of DNA extracted after melanin removal by hydrogen peroxide method was 59.59 μg/L, substantially higher than that of DNA extracted without melanin removal (30.3 μg/L, P=0.001). The A260/ A280 of DNA extracted after melanin removal by hydrogen peroxide was between 1.8 and 2.0 in all cases, and the A260/ A230 was above 2.0 in sixteen cases, suggesting high purity of DNA. However, the DNA extracted without removing the melanin showed poor purity, with A260/ A280 below 1.8 in eight cases and A260/ A230 below 2.0 in sixteen cases. Conclusions:Warm hydrogen peroxide showed the least melanin residue, superior HE staining and a minimal effect on DNA purity/quality compared to the other three methods. It thus appears most suitable for PCR, NGS and other molecular detection. Melanin removal with trichloroisocyanuric acid after immunohistochemical staining has the least melanin residual, and thus could be the most convenient and efficient. However, it is noted that the efficacy of the same depigmentation method varies with different antibodies. Therefore, the optimal depigmentation method should be selected based on the specific markers of interest.

RNA methylation is a common epigenetic post-transcriptional modification with various patterns,such as N1-methyladenosine(m1A),5-methylcytidine(m5C)and N6-methyladenosine(m6A).RNA methylation can perform specific biological functions in corresponding targets,dynamical and reversible,thus bridging exogenous environmental factors and different disease outcomes.Exogenous chemicals can induce oxidative stress,inflammation,autophagy and cell cycle disorders,which are reg-ulated by specific RNA methylation modifications and bring about epigenetic toxic effects.These altera-tions act as new key molecular events during cellular senescence and the development of aging and age-related disorders.The correlation between RNA methylation and cell senescence will provide a new line of thought for prevention of and interventions in aging.

Objective:To investigate the short-term outcome of transoral robotic thyroidectomy.Methods:This is a retrospective case series study. The clinicopathologic characteristics and postoperative results of 107 patients who underwent transoral robotic thyroidectomies in the Department of Thyroid and Breast Surgery of the 960 th Hospital of People′s Liberation Army from May 2020 to August 2023 were retrospectively analyzed. There were 12 males and 95 females, with an age of (31.8±9.4) years (range: 11 to 55 years), including 20 benign tumors and 87 thyroid papillary carcinoma. Postoperative follow-up was carried out through returning visit and telephone, mainly to observe the recovery of postoperative complications, cosmetic effects and recurrence results. Results:All transoral robotic thyroidectomy was successfully completed without conversion to open surgery. The tumor size of thyroid papillary carcinoma patients was (5.6±2.7) mm (range: 2 to 15 mm). Furthermore, central cervical lymph node metastasis was found in 45 cases. The number of central cervical lymph nodes retrieved and metastasized ( M(IQR)) were 11 (8) (range: 3 to 26) and 1 (3) (range: 0 to 13), respectively. There was no recurrent laryngeal nerve injury and permanent hypoparathyroidism. The transient hypoparathyroidism after surgery was 8 cases. Other complications occurred as follows: postoperative infection ( n=1), left submandibular perforation ( n=1), skin scald ( n=1), and perioral numbness ( n=1), oral tear ( n=2). The postoperative stay was 6 (2) days (range: 3 to 11 days). No local lymph node recurrence or metastasis occurred after a follow-up of (22.6±10.0) months (range: 1.0 to 37.4 months). All patients were satisfied with the postoperative cosmetic results, the aesthetic effect score was 9.3 (0.2) (range: 8.4 to 9.6) one month after surgery. Conclusion:For highly screened patients with early thyroid cancer, experienced surgeons can perform a transoral robotic thyroidectomy that has excellent cosmetic results.

Objective:To investigate the short-term outcome of transoral robotic thyroidectomy.Methods:This is a retrospective case series study. The clinicopathologic characteristics and postoperative results of 107 patients who underwent transoral robotic thyroidectomies in the Department of Thyroid and Breast Surgery of the 960 th Hospital of People′s Liberation Army from May 2020 to August 2023 were retrospectively analyzed. There were 12 males and 95 females, with an age of (31.8±9.4) years (range: 11 to 55 years), including 20 benign tumors and 87 thyroid papillary carcinoma. Postoperative follow-up was carried out through returning visit and telephone, mainly to observe the recovery of postoperative complications, cosmetic effects and recurrence results. Results:All transoral robotic thyroidectomy was successfully completed without conversion to open surgery. The tumor size of thyroid papillary carcinoma patients was (5.6±2.7) mm (range: 2 to 15 mm). Furthermore, central cervical lymph node metastasis was found in 45 cases. The number of central cervical lymph nodes retrieved and metastasized ( M(IQR)) were 11 (8) (range: 3 to 26) and 1 (3) (range: 0 to 13), respectively. There was no recurrent laryngeal nerve injury and permanent hypoparathyroidism. The transient hypoparathyroidism after surgery was 8 cases. Other complications occurred as follows: postoperative infection ( n=1), left submandibular perforation ( n=1), skin scald ( n=1), and perioral numbness ( n=1), oral tear ( n=2). The postoperative stay was 6 (2) days (range: 3 to 11 days). No local lymph node recurrence or metastasis occurred after a follow-up of (22.6±10.0) months (range: 1.0 to 37.4 months). All patients were satisfied with the postoperative cosmetic results, the aesthetic effect score was 9.3 (0.2) (range: 8.4 to 9.6) one month after surgery. Conclusion:For highly screened patients with early thyroid cancer, experienced surgeons can perform a transoral robotic thyroidectomy that has excellent cosmetic results.

Uveal melanoma (UM) poses a significant lethality, with approximately 50% of those developing metastases surviving less than one year. In the progression of UM, vasculogenic mimicry (VM) induced by hypoxia plays a pivotal role, which also partially explains the resistance of UM to anti-angiogenic therapies. Nevertheless, the crucial molecular mechanisms underlying VM in the progression of UM remain unclear. We identified ubiquitin conjugating enzyme E2 G2 (UBE2G2) as a critical suppressor through transcriptomic sequencing and metastasis correlation screening. In UM, hypoxia-induced VM and metastasis are markedly exacerbated by UBE2G2 knockdown and significantly alleviated by its overexpression. Mechanistically, UBE2G2 directly binds to galectin 3 binding protein (LGALS3BP) and forms a complex with the E3 ubiquitin ligase tripartite motif containing 38 (TRIM38), facilitating ubiquitination-mediated degradation of LGALS3BP at the K104 residue. Furthermore, UBE2G2 inhibits oncogenic phenotypes by inactivating intracellular PI3K/AKT signaling and reprogramming the tumor microenvironment. Therefore, targeting intercellular and intracellular molecular mechanisms of the hypoxia-UBE2G2-LGALS3BP axis may contribute to developing various therapeutic strategies for UM.

Objective:To investigate the effect of horticultural therapy on social function of elderly inpatients with cognitive dysfunction.Methods:Ninety inpatients with cognitive dysfunction who met the diagnostic criteria of ICD-10 were randomly divided into horticultural research group and control group, with 45 patients in each group.Finally, totally 82 completed the test with 41 in each group.Patients in the two groups were both given drug therapy according to clinical symptoms.At the same time, patients in the research group were given horticultural therapy 3 times a week, 90 minutes each section, for 12 weeks.Scale of social function in psychosis inpatients(SSPI) was used to assess the social function of the patients in the two groups were at baseline, 4, 8 and 12 weekends of the treatment.SPSS 25.0 software was used to conduct chi-square test and independent sample t-test for demographic data of the two groups, and repeated measures ANOVA was performed for SSPI score. Results:The results of repeated measure analysis of variance showed that the group main effect ( F=7.226, F=13.428, F=24.817, F=19.793) and interaction effect between time and group were all statistically significant ( F=29.644, F=42.937, F=53.246, F=67.215)(all P<0.01) in daily living ability (factor Ⅰ), mobility and interaction (factor Ⅱ), social activity skills score (factor Ⅲ) and total score.Simple effect analysis showed there were no statistically significant differences in each factor score and total score between the two groups at the baseline ( P>0.05), and there were statistically significant differences in each factor scores and total score at the end of the 8th weekend ((9.95±2.41), (10.39±3.38), (6.56±3.24), (26.90±7.88) vs (8.10±2.45), (6.88±3.48), (2.81±2.50), (17.78±6.96))and 12th weekend((10.27±2.16), (11.61±3.07), (7.88±3.08), (29.76±7.40) vs (7.56±2.41), (5.78±3.21), (2.34±2.02), (15.68±6.24)) (all P<0.01). The pair-to-pair comparison within the group showed that the differences in factor Ⅱ, factor Ⅲ and total score of the research group at the end of the 12th weekend ((11.61±3.07), (7.88±3.08), (29.76±7.40)) were higher than those at the baseline((8.59±3.93), (4.56±3.32), (22.02±8.35)), 4th weekend((9.07±4.14), (5.12±3.35), (23.59±8.51)), and 8th weekend((10.39±3.38), (6.56±3.24), (26.90±7.88))(all P<0.05). Factor Ⅰ (10.27±2.16) showed a statistically significant difference compared with the baseline (8.88±2.65) and 4th weekend (9.39±2.63)(both P<0.05). All the scores showed an upward trend. Conclusion:Horticultural therapy can improve the social function of elderly inpatients with cognitive impairment.

Transcriptional phenotypic drug discovery has achieved great success,and various com-pound perturbation-based data resources,such as connectivity map(CMap)and library of inte-grated network-based cellular signatures(LINCS),have been presented.Computational strategies fully mining these resources for phenotypic drug discovery have been proposed.Among them,the fundamental issue is to define the proper similarity between transcriptional profiles.Tra-ditionally,such similarity has been defined in an unsupervised way.However,due to the high dimensionality and the existence of high noise in high-throughput data,similarity defined in the tra-ditional way lacks robustness and has limited performance.To this end,we present DrSim,which is a learning-based framework that automatically infers similarity rather than defining it.We evalu-ated DrSim on publicly available in vitro and in vivo datasets in drug annotation and repositioning.The results indicated that DrSim outperforms the existing methods.In conclusion,by learning tran-scriptional similarity,DrSim facilitates the broad utility of high-throughput transcriptional pertur-bation data for phenotypic drug discovery.The source code and manual of DrSim are available at https://github.com/bm2-lab/DrSim/.

The treatment of hepatobiliary malignant tumor is characterized by the coexistence of multiple treatment methods and multiple disciplines. In order to evaluate the clinical efficacy of different treatment measures or multiple treatment combinations, and to promote the standardized development of comprehensive treatment patterns for hepatobiliary malignant tumor, the Hepatobiliary Center of the First Affiliated Hospital of Nanjing Medical University constructs the registry and follow-up database in hepatobiliary tumor patients based on the information-based platform of the hospital, which will help guide clinicians to make scientific decisions and improve the level of clinical diagnosis and treatment. This study describes the framework design, function modules, data acquisition process and quality control of the database of hepatobiliary malignant tumor. Based on the observational bidirectional cohort study design, the previous clinical data can be sorted to match the current database, on the other hand, the clinical data can be prospectively collected including basic information, admission evaluation, surgical information and postoperative situation, comprehensive treatment measures, regular reexaminations and long-term follow-up, etc. The data quality control system can be improved by formulating standardized operation procedures, regularly personnel training and full-process data management plans. This database will provide high-quality real-world data for clinicians, researchers, and guideline experts, and then provide high-level medical evidence for the standardized development of comprehensive treatment patterns of hepatobiliary malignancies.

Objective:To investigate mentor-mentee dual evaluation of the current status of mentor competency of clinical and translational research mentors, and provide the basis for mentor training focused on the mentor competences.Methods:A total of 121 clinical and translational research mentors and 170 mentees from Peking Union Medical College Hospital were enrolled. The Chinese version of the Mentor Competency Assessment (MCA) questionnaire was used to evaluate the mentor competency, including maintaining effective communication, aligning expectations, assessing understanding, fostering independence, addressing diversity, promoting professional development total 6 parts with 26 sub-items. The Likert scale was used to quantitatively evaluate the clinical and translational research mentor competency by mentor-mentee dual evaluations. And the composition and training needs of clinical and translational research mentors were investigated. SPSS 25.0 was used for t-test. Results:Seventy percent (119/170) of mentees considered the mentor guidance was very helpful, and 78.5% (95/121) of the mentors considered it necessary to carry out the mentor training. The mentee evaluation of mentor competency was significantly higher than that of mentor self-evaluation [total score (162.35±23.59) vs. (154.80±19.81), P < 0.01]. And the excellent rate of 26 sub-items by mentees and mentors were 100.0%(170/170) and 46.3%(56/121) respectively. The mentors and mentees shared the agreement of the strengths on trust-based relationship and encouraging mentees, and weaknesses on taking into account the possible prejudices in mentor-mentee relationship. Conclusion:The clinical and translational research mentors have already had good competences, but mentor training is still highly warranted. It's expected that to carry out targeted mentor training and assessment according to the mentor's competences will help to improve the construction of the medical talents training system.