Frostbite is the most common cold injury and is caused by both immediate cold-induced cell death and the gradual development of localized inflammation and tissue ischemia. Delayed healing of frostbite often leads to scar formation, which not only causes psychological distress but also tends to result in the development of secondary malignant tumors. Therefore, a rapid healing method for frostbite wounds is urgently needed. Herein, we used a mouse skin model of frostbite injury to evaluate the recovery process after frostbite. Moreover, single-cell transcriptomics was used to determine the patterns of changes in monocytes, macrophages, epidermal cells, and fibroblasts during frostbite. Most importantly, human-induced pluripotent stem cell (hiPSC)-derived skin organoids combined with gelatin-hydrogel were constructed for the treatment of frostbite. The results showed that skin organoid treatment significantly accelerated wound healing by reducing early inflammation after frostbite and increasing the proportions of epidermal stem cells. Moreover, in the later stage of wound healing, skin organoids reduced the overall proportions of fibroblasts, significantly reduced fibroblast-to-myofibroblast transition by regulating the integrin α5β1-FAK pathway, and remodeled the extracellular matrix (ECM) through degradation and reassembly mechanisms, facilitating the restoration of physiological ECM and reducing the abundance of ECM associated with abnormal scar formation. These results highlight the potential application of organoids for promoting the reversal of frostbite-related injury and the recovery of skin functions. This study provides a new therapeutic alternative for patients suffering from disfigurement and skin dysfunction caused by frostbite.
Animals ; Organoids/metabolism* ; Mice ; Humans ; Wound Healing ; Frostbite/metabolism* ; Skin/pathology* ; Induced Pluripotent Stem Cells/cytology* ; Cicatrix/pathology* ; Fibroblasts/metabolism* ; Disease Models, Animal ; Mice, Inbred C57BL ; Extracellular Matrix/metabolism* ; Male

Objective To study the expression of N6-methyladenosine(m6A)methyltransferase-like protein 16(METTL16),Ankyrin repeat and SOCS box containing protein 4(ASB13)in elderly breast cancer tissues and their relationship with clinicopathological features and prognosis.Methods 124 elderly patients with breast cancer diagnosed and treated in the Zibo 148 Hospital from January 2019 to January 2021 were selected.Immunohistochemistry(IHC)was used to detect the expression of METTL16 and ASB13 proteins in tissues.Real time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression of METTL16 mRNA and ASB13 mRNA in tissues.Pearson correlation was used for correlation analysis.Survival analysis used Kaplan-Meier curves and Log Rank tests to compare the prognostic differences among patients with different METTL16 and ASB13 expression levels.COX regression analysis was used to analyze the prognostic factors of elderly patients with breast cancer.Results The positive rate of METTL16 in cancer tissue(70.97%)was higher than that in adjacent tissues(8.06%),while the positive rate of ASB13(22.58%)was lower than that in adjacent tissues(72.58%),the differences were statistically significant(χ2=102.642,62.146,all P<0.001).METTL16 mRNA,Snail mRNA and N-cadherin(N-cd)mRNA were higher in cancer tissues than in adjacent tissues,while ASB13 mRNA was lower in adjacent tissues,the differences were statistically significant(t=40.498～48.624,all P<0.001).METTL16 mRNA was positively correlated with Snail mRNA and N-cd mRNA in cancer tissue(r=0.712,0.669,all P<0.001),while ASB13 mRNA was negatively correlated with Snail mRNA and N-cd mRNA(r=-0.734,-0.759,all P<0.001).Compared with TNM stage Ⅰ-Ⅱ and patients with high to medium differentiation,patients with TNM stage Ⅲ and low differentiation had higher positive rate of METTL16 in cancer tissue and lower positive rate of ASB13,the differences were statistically significant(χ2=5.340～10.370,all P<0.05).The 3-year progression free survival rate(73.86%)of the METTL16 positive group was lower than that of the METTL16 negative group(88.89%),the 3-year progression free survival rate(92.86%)of the ASB13 positive group was higher than that of the ASB13 negative group(73.96%),and the differences were statistically significant(Log Rank χ2=4.483,4.882,all P<0.005).METTL16 positive,TNM stage Ⅲ,poor differentiation were risk factors affecting the prognosis of elderly breast cancer,and ASB13 positive was protective factors(Wald χ2=5.056～9.267,all P<0.001).Conclusion METTL16 expression is up-regulated and ASB13 expression is down-regulated in elderly breast cancer,both of which are related to the expression of invasion and metastasis genes,and are new prognostic markers for elderly breast cancer.

Objective To analyze the effects of different traction sites and directions on the maxilla and upper dentition when using clear aligners combined with protraction for the treatment of maxillary deficiency.Methods A three-dimensional(3D)finite element model including the zygomaticomaxillary complex,maxillary dentition,and clear aligners was constructed.The models were divided into Group 1(traction hook at the distal of the lateral incisor)and Group 2(traction hook at the distal of the canine).Each group was analyzed under four loading conditions with protraction angles of 0°,10°,20°,and 30° relative to the occlusal plane.A unilateral protraction force of 500 g was applied.The differences in stress distribution and displacement of the maxillary bone and dentition under different loading conditions were analyzed.Results When the protraction angle was 30°,both groups showed forward and downward displacement of the maxilla,while other angles resulted in counterclockwise rotation.Under the same protraction direction,the total displacement of the maxilla and displacements in all directions in Group 2 were greater than those in Group 1.The upper central incisors in Group 1 showed lingual displacement,which increased with the protraction angle.The maxillary dentition in Group 2 showed forward displacement,with the minimum total and sagittal displacements at a protraction angle of 30°.Stress concentration was mainly observed in the zygomaticomaxillary suture and anterior alveolar bone regions in both groups,decreasing as the protraction angle increased.Conclusions Clear aligners combined with protraction can be applied to skeletal Class Ⅲ patients with mild maxillary deficiency.When the protraction site is located at the distal of the canine with a 30° downward and forward angle to the occlusal plane,the maxilla can achieve ideal forward and downward displacement with the minimum labial movement of the upper anterior teeth.

Objective To study the expression of N6-methyladenosine(m6A)methyltransferase-like protein 16(METTL16),Ankyrin repeat and SOCS box containing protein 4(ASB13)in elderly breast cancer tissues and their relationship with clinicopathological features and prognosis.Methods 124 elderly patients with breast cancer diagnosed and treated in the Zibo 148 Hospital from January 2019 to January 2021 were selected.Immunohistochemistry(IHC)was used to detect the expression of METTL16 and ASB13 proteins in tissues.Real time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression of METTL16 mRNA and ASB13 mRNA in tissues.Pearson correlation was used for correlation analysis.Survival analysis used Kaplan-Meier curves and Log Rank tests to compare the prognostic differences among patients with different METTL16 and ASB13 expression levels.COX regression analysis was used to analyze the prognostic factors of elderly patients with breast cancer.Results The positive rate of METTL16 in cancer tissue(70.97%)was higher than that in adjacent tissues(8.06%),while the positive rate of ASB13(22.58%)was lower than that in adjacent tissues(72.58%),the differences were statistically significant(χ2=102.642,62.146,all P<0.001).METTL16 mRNA,Snail mRNA and N-cadherin(N-cd)mRNA were higher in cancer tissues than in adjacent tissues,while ASB13 mRNA was lower in adjacent tissues,the differences were statistically significant(t=40.498～48.624,all P<0.001).METTL16 mRNA was positively correlated with Snail mRNA and N-cd mRNA in cancer tissue(r=0.712,0.669,all P<0.001),while ASB13 mRNA was negatively correlated with Snail mRNA and N-cd mRNA(r=-0.734,-0.759,all P<0.001).Compared with TNM stage Ⅰ-Ⅱ and patients with high to medium differentiation,patients with TNM stage Ⅲ and low differentiation had higher positive rate of METTL16 in cancer tissue and lower positive rate of ASB13,the differences were statistically significant(χ2=5.340～10.370,all P<0.05).The 3-year progression free survival rate(73.86%)of the METTL16 positive group was lower than that of the METTL16 negative group(88.89%),the 3-year progression free survival rate(92.86%)of the ASB13 positive group was higher than that of the ASB13 negative group(73.96%),and the differences were statistically significant(Log Rank χ2=4.483,4.882,all P<0.005).METTL16 positive,TNM stage Ⅲ,poor differentiation were risk factors affecting the prognosis of elderly breast cancer,and ASB13 positive was protective factors(Wald χ2=5.056～9.267,all P<0.001).Conclusion METTL16 expression is up-regulated and ASB13 expression is down-regulated in elderly breast cancer,both of which are related to the expression of invasion and metastasis genes,and are new prognostic markers for elderly breast cancer.

Objective To analyze the effects of different traction sites and directions on the maxilla and upper dentition when using clear aligners combined with protraction for the treatment of maxillary deficiency.Methods A three-dimensional(3D)finite element model including the zygomaticomaxillary complex,maxillary dentition,and clear aligners was constructed.The models were divided into Group 1(traction hook at the distal of the lateral incisor)and Group 2(traction hook at the distal of the canine).Each group was analyzed under four loading conditions with protraction angles of 0°,10°,20°,and 30° relative to the occlusal plane.A unilateral protraction force of 500 g was applied.The differences in stress distribution and displacement of the maxillary bone and dentition under different loading conditions were analyzed.Results When the protraction angle was 30°,both groups showed forward and downward displacement of the maxilla,while other angles resulted in counterclockwise rotation.Under the same protraction direction,the total displacement of the maxilla and displacements in all directions in Group 2 were greater than those in Group 1.The upper central incisors in Group 1 showed lingual displacement,which increased with the protraction angle.The maxillary dentition in Group 2 showed forward displacement,with the minimum total and sagittal displacements at a protraction angle of 30°.Stress concentration was mainly observed in the zygomaticomaxillary suture and anterior alveolar bone regions in both groups,decreasing as the protraction angle increased.Conclusions Clear aligners combined with protraction can be applied to skeletal Class Ⅲ patients with mild maxillary deficiency.When the protraction site is located at the distal of the canine with a 30° downward and forward angle to the occlusal plane,the maxilla can achieve ideal forward and downward displacement with the minimum labial movement of the upper anterior teeth.

The aryl hydrocarbon receptor(AhR),also known as dioxin receptor,is a ligand-dependent transcription factor.Because of its important role in occurrence and development of cancer,it has been widely studied.AhR is now considered to be an important regulator of host-environment interactions in immune and inflammatory responses and is involved in pathogenesis of many skin diseases.Because AhR is highly expressed in all types of skin cells and regulates many genes that are critical to skin function,it has the potential to be a new target for the treatment of inflammatory skin diseases.This paper presents and analyzes research findings on the relationship between AhR and inflammatory skin diseases to help accelerate the development of new drugs.

Objective To investigate the key elements for model establishment and determine the evaluation indicators of animal models for pelvic inflammatory disease(PID),providing a reference for improving modelling methods and optimizing the application of PID animal models.Methods The search query"Pelvic Inflammatory Disease"AND"Animal Model"OR"Rat"OR"Mouse"OR"Guinea Pig"OR"Rabbit"OR"Dog"OR"Pig"was used to retrieve relevant literature on PID animal models published from 2013 to 2023 in China Knowledge Network Infrastructure(CNKI),Wanfang,and PubMed databases.The studies were analyzed and categorized based on experimental animal types,modelling methods,modelling cycles,detection indicators,positive control drugs,and administration duration.A database was established for statistical analysis.Results A total of 214 research articles on PID animal models meeting the inclusion criteria were identified.The most commonly used model animals are Sprague Dawley(SD)rats,followed by Wistar rats.The most frequently employed modelling method is a combination of mechanical injury and bacterial infection,followed by the phenol mucilage method.The most common modelling cycles for acute pelvic inflammatory disease(APID)and chronic pelvic inflammatory disease(CPID)/sequelae of pelvic inflammatory disease(SPID)are 8 to 14 days,while for PID models without specific staging,the cycles are 7 days.High-frequency detection methods and indicators include histopathological observation using hematoxylin-eosin staining,enzyme-linked immunosorbent assay(ELISA)for serum-related indicators,morphological changes of tissues observed with the naked eye,and immunohistochemical detection of related protein expression in uterine tissues,and pathological scoring.The most frequently used positive control drugs are Fuke Qianjin Tablets,followed by Jingangteng Capsules.The most common administration duration for APID is 7 days,and for CPID/SPID models,it ranges from 15 to 21 days.Conclusion Currently,SD rats and Wistar rats are commonly used as experimental animals for PID models.The dual modelling method of mechanical injury combined with mixed bacterial infection aligns closely with clinical pathogenesis and can be used to establish a PID model that simulates postoperative uterine cavity infection.Depending on the research objectives,different positive drugs and detection indicators should be selected for comprehensive evaluation.Most existing PID animal model studies are based on western medical diagnosis,with fewer studies focusing on Traditional Chinese Medicine(TCM)syndromes.There is a need to integrate TCM theories of etiology and pathogenesis to construct PID animal models that are more in line with TCM clinical symptoms.

Objective To comprehensively analyze the reported preparation methods for animal models of perimenopausal syndrome (PS), to compare the advantages and disadvantages of various preparation elements and detection indexes, so as to provide useful references for the optimization of the relevant animal models as well as the standardization of their application in the efficacy evaluation of new drugs.MethodsIn this paper, literature research methods were applied using "perimenopausal syndrome" as the subject term. The publication period of the literature was limited to January 2016 to February 2023. Relevant literature on the preparation of PS animal models was retrieved from databases such as China National Knowledge Infrastructure, Wanfang database, and PubMed. After screening the experimental literature that met the inclusion and exclusion criteria, detailed information on experimental animal strains, modeling methods, duration of drug administration, positive drugs, detection indexes and other relevant information were collected. After the above information was standardized, the PS animal model database was established using Excel 2010 software. The model preparation elements and evaluation indexes were summarized systematically, and the statistical results were processed and analyzed using Excel 2010 software.Results A total of 247 articles were screened. SD rats (164 times, 65.86%) and Wistar rats (35 times, 14.06%) were often used to prepare PS animal models. Bilateral ovariectomy (139 times, 53.87%) and natural aging (43 times, 16.80%) were chosen as modeling methods. The ages of rats used for modeling ranged from 7 weeks to 18 months, with 3-month-old rats (22 times, 21.78%) being the most common. The detection indexes were comprehensively evaluated from multiple perspectives, including serum biochemistry, vaginal exfoliated cell smear, histomorphology, general observation, behavioral observation, and organ tissue protein immunoblotting. Western medical evaluation indexes were commonly used to test the successful preparation of models, with vaginal exfoliated cell smears being the most frequently used method (125 times, 85.04%). A model was considered successfully prepared when estrous cycle disorder or irregularity was observed. Some literature also determined modeling success by detecting a significant decrease in serum estradiol levels (5 times, 3.04%). Traditional Chinese medicine (TCM) syndrome evaluation often used a combination of Chinese and Western medical evaluation indexes for comprehensive evaluation, with researchers determining the TCM syndrome through vaginal exfoliated cell smears supplemented by general observation (3 times, 2.04%).Conclusion There are many methods for preparing PS animal models, but there are still significant differences in the selection of animal species, age, criteria for successful modeling, and TCM syndrome evaluation in the related literature.

The incidence of augmented renal clearance (ARC) in intensive care patients (ICU) is exceptionally high, and these patients are often co-morbid with infection. The occurrence of ARC will significantly increase the clearance rate of antibiotics, making it difficult for conventional doses to reach effective therapeutic concentrations and affect the patient's anti-infective treatment effect and prognosis. It can be seen that it is crucial to formulate a reasonable dosing regimen for ICU patients with ARC. Regrettably, few reports in China about the adjustment strategy of antibiotic dosing regimens for ARC patients. Therefore, this article reviews the domestic and foreign literature for reference to provide evidence for medical personnel to adjust the dose of antibacterial drugs for such patients.