Objective: This study aimed to evaluate the effectiveness of podiatrists in preventing diabetic foot ulcers (DFUs) in China. Method: The study was a prospective investigation. A total of 300 patients were enrolled from May 2016 to May 2018 in Handan Central Hospital, China. All patients who participated in this study had been diagnosed with type 2 diabetes, according to the International Classification of Diseases (ICD-10). All participants underwent our survey, which included basic patient data and information about DFUs. The patients were followed for one year, during which time they received appropriate intervention from podiatrists, including lifestyle guidance, callus resection, tinea grinding and ingrown nail correction. At the end of the year all the patients were surveyed again. The data before and after the year were statistically compared. Results: The results showed that the incidence of DFUs in patients with diabetes was significantly decreased after one year of intervention from podiatrists (20.7% versus 6.7%, p<0.001). Additionally, there was a negative correlation between the number of intervention visits and the number of DFU occurrences (Spearman correlation coefficient: –0.496, p<0.001). Furthermore, we found that 68 patients with a history of DFUs or amputation had an obviously reduced incidence of DFUs after intervention by a podiatrist (89.7% versus 27.9%, p<0.001). We also investigated other foot risk factors in all participants, such as limb neuropathy (76.3%), lower extremity vascular disease (65.7%) and foot paralysis (43.7%). Conclusion: The results of this study help in understanding the situation of patients with diabetes in China and to prove that standardised podiatrist intervention has an important role in inhibiting the occurrence and development of DFUs.

Background Venous leg ulcers (VLUs) are the most severe manifestation of chronic venous disease, with long healing time and a high recurrence rate. It imposes a heavy burden on patients, their families, and the health care system. Chronic inflammation triggered by sustained venous hypertension is now recognized as the hallmark of chronic venous disease. The anti-inflammatory effect of pentoxifylline may offer a promising avenue to treat VLUs. However, current evidence of pentoxifylline for VLUs is relatively small and of low quality. The aim of this study is to evaluate the efficacy and safety of pentoxifylline for VLUs in the Chinese population. Methods This is a randomized, double-blinded, double-dummy, multi-center, placebo-controlled clinical trial. A total of 240 patients will be randomized to receive pentoxifylline (400 mg, twice daily) or placebo for 24 weeks. All participants will receive diosmin treatment and standard care of VLUs and other comorbidities. The primary outcome is the difference in the wound healing rate within 12 weeks between pentoxifylline and placebo. Secondary outcomes include (1) percent wound size changes at 12 weeks, (2) the levels of TNF-α and IL-6, (3) venous clinical severity score and chronic venous insufficiency quality of life score, and (4) ulcer recurrence within 24 weeks. Discussion This study would evaluate the efficacy and safety of pentoxifylline for VLUs in the Chinese population. If confirmed, it wound offer another effective and safe therapeutic option for treatment of VLUs. Trial registration The trial was registered at the Chinese Clinical Trial Registry (No. ChiCTR-2100053053). Registered on 10 November, 2021, https://www.chictr.org.cn/showproj.aspx?proj=137010

Aims: This study aims to investigate the function of positive feedback loops involving noncoding RNA in diabetic wound healing. Methods: We developed a mouse diabetic wound model to confirm that hyperglycemia can impair wound healing. We also used an in vitro keratinocyte model in high-glucose conditions to investigate the mechanism of delayed wound healing. Results: MALAT1 was decreased in diabetic mouse wound tissue and can promote keratinocyte biological functions. MALAT1 could bind to miR-106a-5p to modulate the expression of ZNF148, a target gene of miR-106a-5p. Surprisingly, ZNF148 bound to a region in the MALAT1 promoter to stimulate gene expression. Conclusion: ZNF148-activated MALAT1 increases ZNF148 expression by competitively binding miR-106a-3p, generating a positive feedback loop that enhances keratinocyte function.

Objective:To study the inducement and location of diabetes foot ulcer, it provide a basis for the prevention and control of diabetes foot.Methods:Data of 230 inpatients with diabetic foot admitted to wound Repair Center of Liyuan Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from January 2019 to September 2021 were retrospectively analyzed. Collected the information includes the patient′s gender, age, place of residence, underlying diseases, whether other diseases were present at the time of onset, the cause and location of foot ulcers, and risk factors. Data were compared by χ2 test and Kruskal-Wallis H test. Results:(1) 112 cases of foot ulcer caused by daily life and health care behavior, accounting for 48.70% of the total number; there were 69 cases of ulcers without obvious inducements, accounting for 30.00%; 49 cases (21.30%) were caused by itching, dry chapped skin, edema, corns and other complications. Foot ulcers in 178 cases were recurrent, accounted for 77.39%, in which 98 cases caused by inadequate health and daily life behavior, accounting for 42.61%. The inducement of foot ulcers had a greater impact on recurrence than initial onset, the difference was statistically significant (χ2=12.759, P< 0.05). (2) Among the 230 patients with diabetic foot ulcer, 119 cases (51.74%) of the patients with recurrent ulcer were the dorsum of the foot, followed by 40 cases (17.39%) of the plantar, 19 cases (8.26%) of the dorsum and plantar. The majority of patients with recurrent ulcers suffered from the dorsum of the foot, there was statistically significant difference in the location of onset between initial and recurrent ulcers (χ2=12.349, P< 0.05). (3) 112 cases were caused by inappropriate daily life and health care behavior, and 77 cases (33.48%) occurred in the dorsum of the foot. Compared different ulcer site with incentive, the difference was statistically significant (χ2=25.934, P<0.001). (4)198 cases (86.09%) of ulcers occurred on the forefoot, followed by 21 cases (9.13%) of the hind foot, 8 cases (3.48%) in two or more areas, only 3 cases of patients′ ulcers occurred in the midfoot, accounting for 1.30%. Comparing the incidence site and location of ulcers, ulcers often occured at the dorsum of the forefoot, with a statistically significant difference (χ2=23.530, P<0.05).Conclusion:Diabetes foot ulcer is more recurrent, the recurrence of diabetes foot ulcer is mostly located in the forefoot, and the dorsum of the foot is more than the sole of the foot. The inducement of recurrence is mostly inappropriate daily life and health care behavior, which provides a basis for the prevention of diabetes foot recurrence.

Abstract Diabetic foot ulcers (DFUs) are associated with complex pathogenic factors and are considered a serious complication of diabetes. The potential mechanisms underlying DFUs have been increasingly investigated. Previous studies have focused on the three aspects of diabetic peripheral vascular disease, neuropathy, and wound infections. With advances in technology, researchers have been gradually conducting studies using immune cells, endothelial cells, keratinocytes, and fibroblasts, as they are involved in wound healing. It has been reported that the upregulation or downregulation of molecular signaling pathways is essential for the healing of DFUs. With a recent increase in the awareness of epigenetics, its regulatory role in wound healing has become a much sought‐after trend in the treatment of DFUs. This review focuses on four aspects involved in the pathogenesis of DFUs: physiological and pathological mechanisms, cellular mechanisms, molecular signaling pathway mechanisms, and epigenetics. Given the challenge in the treatment of DFUs, we are hopeful that our review will provide new ideas for peers.

Diabetic foot ulcers (DFUs) are associated with complex pathogenic factors and are considered a serious complication of diabetes. The potential mechanisms underlying DFUs have been increasingly investigated. Previous studies have focused on the three aspects of diabetic peripheral vascular disease, neuropathy, and wound infections. With advances in technology, researchers have been gradually conducting studies using immune cells, endothelial cells, keratinocytes, and fibroblasts, as they are involved in wound healing. It has been reported that the upregulation or downregulation of molecular signaling pathways is essential for the healing of DFUs. With a recent increase in the awareness of epigenetics, its regulatory role in wound healing has become a much sought-after trend in the treatment of DFUs. This review focuses on four aspects involved in the pathogenesis of DFUs: physiological and pathological mechanisms, cellular mechanisms, molecular signaling pathway mechanisms, and epigenetics. Given the challenge in the treatment of DFUs, we are hopeful that our review will provide new ideas for peers.

Purpose:Diabetic wound is a highly prevalent and refractory disease. Extensive studies have confirmed that keratinocytes and macrophages play an important role in the process of wound healing. Additionally, exosomes are regarded as a vital intercellular communication tool. This study aimed to investigate the role of human keratinocyte-derived exosomal MALAT1 in the treatment of diabetic wound by influencing the biological function of macrophages.Methods:We mainly assessed the function of MALAT1 on the biological changes of macrophages, and the expression of MALAT1 in the keratinocyte-exosomes analyzed by quantitative real-time polymerase chain reaction (RT-qPCR). The downstream interaction between RNAs or proteins was assessed by mechanistic experiments. Besides, we evaluated the effects of human keratinocyte-derived exosomal MALAT1 on diabetic wound healing in vivo to verify in vitro results.Results:We demonstrated that human keratinocyte-derived exosomal MALAT1 enhanced the biological functions of high glucose-injured macrophages, including phagocytosis, converting to a pro-healing phenotype and reducing apoptosis. Mechanistically, MALAT1 accelerated the expression of MFGE8 by competitively binding to miR-1914-3p, thereby affecting the function of macrophages and the signal axis of TGFB1/SMAD3, and finally promoting the healing of diabetic wounds. Human keratinocyte-derived exosomal MALAT1 might promote collagen deposition, ECM remodeling, and expression of MFGE8, VEGF, and CD31 but reduce the expression of TGFB and SMAD3 in an in vivo model of diabetic mice wounds, which accelerated diabetic wound healing and restored its function.Conclusion:The current study revealed that human keratinocyte-derived exosomal MALAT1 would suppress miR-1914-3p to activate MFGE8 and eventually promote wound healing by enhancing macrophage phagocytosis, converting to a pro-healing phenotype and reducing apoptosis. It proposed that keratinocyte-derived exosomes might have the capacity to serve as a new method for the clinical treatment of diabetic wound.

本文着重阐述了2021年糖尿病足相关基础研究和临床治疗策略的研究进展,以期为治疗糖尿病足提供借鉴帮助.

Diabetic foot is a serious complication of diabetes, and incidence rate is increasing year by year, which is the main reason leading to amputation. The pathogenesis of diabetic foot is complex and the clinical manifestations are changeable. The treatment of diabetes foot is difficult, long time and high cost, causing a huge psychological and economic burden to patients. In recent years, the research on diabetic foot has been developing, and some new diagnostic strategies and therapies have been put forward. This paper reviews the related definition, classification, pathogenesis, diagnosis and treatment progress of diabetic foot, and prospects the future diagnosis and treatment mode of diabetic foot.

Objective:To investigate the effects of long non-coding RNA lymphoenhancer binding factor 1 antisense RNA1 (LEF1-AS1) gene silencing on the proliferation, migration and angiogenesis of human umbilical vein endothelial cells (HUVECs).Methods:HUVECs were obtained from american type culture collection (ATCC). The expression of LEF1-AS1 gene was silenced in huvecs by RNA interference. The cells were divided into control group and experimental group, namely NC-small interfering RNA (siRNA) group and LEF1-AS1-sirna group. The cell counting kit-8 (CCK-8) and Transwell assays were used to detect the changes of cell proliferation and migration, and the effect of LEF1-AS1 on angiogenesis was detected in vitro. The experimental data were expressed as ± s. The independent-sample t-test was used to compare the two groups. Results:RT-qPCR results showed that the relative expression of LEF1-AS1 mRNA in si-LEF1-AS1 group (0.059±0.010) was lower than that in NC group (1.000±0.018), and the difference was statistically significant ( t=40.824, P<0.01). HUVECs had lower absorbance ( A) values (0.103±0.004) in si-LEF1-AS1 group than in NC group (0.129±0.006) at 24 h after treatment. The A values (0.235±0.016) in si-LEF1-AS1 group were also lower than in NC group (0.431±0.018) at 48 h after treatment. The A values in si-LEF1-AS1 group (0.454±0.029) were lower than those in NC group (0.846±0.034) after 72 h of treatment. Transwell migration experiment showed that the relative number of cells migrating to the inferior chamber in SI-LEF1-AS1 group (0.446±0.025) was significantly less than that in NC group (1.000±0.046, t=17.120, P<0.01). The results of angiogenic experiment showed that the length of the adult tube in SI-LEF1-AS1 group (3 152.333±200.959) was significantly shorter than that in NC group (13 263.667±247.823, t=54.891, P<0.01). Conclusion:Silencing LEF1-AS1 inhibits the proliferation, migration and angiogenesis of HUVECs.

Objective:Analyzed the epidemiological characteristics of inpatients with chronic refractory wounds in a wound repair center in Hubei province by reviewing the data of them, in order to provide reference for the formulation of prevention and treatment strategies for the disease.Methods:The case data of inpatients with chronic refractory wounds admitted to the Wound Repair Center of Liyuan Hospital of Tongji Medical College of Huazhong University of Science and Technology from January 2018 to December 2019 were retrospectively analyzed. The collected information includes the patient's gender, living area, occupation, underlying disease, smoking history, wound type, age, hospital stay, hospitalization cost, treatment method (surgical treatment, conservative treatment), clinical outcome, various medical expenses and wound surface detection of pathogenic bacteria. Data were compared by chi-square test and Kruskal-Wallis H test.Results:(1) A total of 2 402 inpatients with chronic refractory wounds were included in this study, including 1 593 males (66.3%) and 809 females (33.7%); there were 1 459 (60.7%) of the urban population and 943 cases (39.3%) of the rural population; the proportion of patients engaged in retirement and manual labor was relatively large, 52.1% and 28.8%, respectively; 74.4% of the patients had diabetes, followed by hypertension (57.5%) and heart disease (31.8%); 52.7% of patients had a history of smoking. (2) Among different wound types, diabetic wounds were the most common, accounting for 62.3% (1 497/2 402); followed by pressure wounds, accounting for 13.9% (334/2 402); arterial wounds accounted for 6.2% (148/2 402); venous wounds accounted for 5.0% (121/2 402), and wounds caused by other reasons accounted for 12.6% (302/2 402). (3) The gender and age distribution of chronic refractory wound patients with different types of wounds were compared, and the differences were statistically significant (χ2=30.564, 28.536; P<0.05); the age group with the highest prevalence rate was 61-80 years old (54.7%), followed by 41-60 years old (29.4%) and over 80 years old (11.8%); 41-60 years old and 61-80 years old with high incidence of diabetic wounds and venous wounds, and 61-80 years old and over 80 years old were the high incidence of pressure wounds and arterial wounds. (4) Among the 2 402 inpatients with chronic refractory wounds, the patients with pressure wounds had the longestaverage hospital stay, which was 31 (19, 35) d, and the patients with arterial wounds and venous wounds had the shortest average hospitalization time, which were 19 (9, 25) d and 19 (11, 26) d, respectively. The average hospitalization cost of patients with arterial wounds was the highest, which was 91 361.5 (17 033.5, 143 090.4) yuan, followed by patients with diabetic wounds, which was 45 008.4 (10 574.0, 57 020.7) yuan. There were statistically significant differences in hospitalization time and hospitalization expenses among patients with different types of chronic refractory wounds (χ2=25.346, 32.460; P<0.05). (5) Among the patients treated by surgery, patients with pressure wounds had the longest hospitalization time 39 (26, 42) d, and patients with arterial wounds had the highest hospitalization costs, which was 118 341.1 (77 151.3, 162 281.1) yuan; among the patients with conservative treatment, other wound patients had the longest hospitalization time, and patients with arterial wounds had the highest hospitalization costs; the hospitalization costs and hospitalization time of patients with different treatment methods were statistically significantly differents (χ2=110.260, 120.765; P<0.05). (6) Among the 2 402 patients, 22.0% (529/2 402) were cured, 76.3% (1 833/2 402) were markedly effective, 1.7% (40/2 402) were not cured, and 12 of them died, accounting for 0.5% of the total number of patients. The hospitalization time of the cured, markedly effective and unhealed patients were 31 (15, 39), 25 (12, 30), and 19 (5, 24) d, respectively, and the hospitalization expenses were 45 360.6 (11 664.3, 56 208.8), 42 824.2 (10 107.1, 52 415.1), 37 017.4 (8 031.3, 31 942.9) yuan, respectively. The hospitalization time of cured patients was longer than that of markedly effective and unhealed patients, and the hospitalization cost was higher than that of markedly effective and unrecovered patients. There were no statistically significant differences in hospitalization time and hospitalization cost among patients with different clinical outcomes (χ2=3.031, 6.780; P>0.05). (7) The total medical expenses of the 2 402 inpatients with chronic refractory wounds were 103 973 107 yuan. Consumables and medicines accounted for a relatively high proportion of various medical expenses, accounting for 42.1% and 23.4%, respectively. Nursing expenses and blood transfusion expenses accounted for a relatively low proportion, at 1.2% and 0.2%, respectively. (8) Among the 2 402 patients, 926 (38.6%) were examined for pathogenic bacteria on the wound surface, of which 681 were positive for pathogenic wounds, with a positive rate of 73.5% (681/926). Among them, Gram-negative bacteria accounted for 50.8% (346/681), mainly Pseudomonas aeruginosa (13.5%), Escherichia coli (11.6%), and Acinetobacter baumannii (6.2%); Gram-positive bacteria accounted for 42.4% (289/681), mainly Staphylococcus aureus (17.8%), Staphylococcus epidermidis (7.2%), and Enterococcus (9.2%); fungi accounted for 6.8% (46/681), mainly Candida albicans (2.5%).Conclusions:The inpatients with chronic refractory wounds are mainly middle-aged and elderly people, and there are more males than females. Diabetic wounds, pressure wounds and vascular wounds are the main causes of the disease. Patients with pressure wounds had the longest average hospital stay, and patients with arterial wounds had the highest average hospitalization costs. The main treatment method is surgery. The unit adopts a multidisciplinary diagnosis and treatment mode for wound treatment, and the healing rate is high. Gram-negative bacteria are the most common pathogens on the wound surface, mainly Pseudomonas aeruginosa, Escherichia coli, and Acinetobacter baumannii. It is necessary to advocate and improve the two-way referral mechanism between wound treatment centers in large general hospitals and community health service centers, and establish wound repair and nursing centers, strengthen publicity and education about the disease, and improve awareness of prevention and treatment.

目的 探讨不同亚型巨噬细胞条件培养基对脐静脉内皮细胞(HUVECs)的生物学功能影响及机制.方法 将巨噬细胞RAW264.7分为M0、M1、M2和si-乳脂球表皮生长因子-8(MFG-E8)组.M0型巨噬细胞不做处理;M1型巨噬细胞采用脂多糖(LPS,100ng/ml)及γ-干扰素(IFN-γ,10ng/ml)联合刺激;M2型巨噬细胞使用白细胞介素(IL)-4(10 ng/ml)诱导;si-MFG-E8组通过小干扰寡核苷酸(siRNA)沉默MFG-E8的表达.诱导48 h后免疫荧光法鉴定巨噬细胞极化,实时定量聚合酶链反应(RT-PCR)验证siRNA敲降效率,并收集各组巨噬细胞条件培养基;酶联免疫吸附试验(ELISA)检测各组巨噬细胞及其条件培养基中MFG-E8的表达;细胞计数试剂盒(CCK-8)检测各组巨噬细胞条件培养基对脐静脉内皮细胞增殖能力的影响;Transwell实验检测各组巨噬细胞条件培养基对脐静脉内皮细胞迁移能力的影响;RT-PCR检测各组的磷脂酰肌醇3激酶(PI3K)、蛋白激酶B(Akt)和雷帕霉素靶蛋白(mTOR)分子的表达水平.组间比较采用非配对t检验.结果 免疫荧光法鉴定巨噬细胞成功极化为M1和M2型.RT-PCR验证与si-NC比较,si-MFG-E8组MFG-E8 mRNA的表达含量明显降低(t=63.070,P＜0.01).ELISA实验结果显示,与MO组比较,M2组巨噬细胞 MFG-E8 明显增高,M1 及 si-MFG-E8 组明显降低(t02=14.050,P02＜0.01;t01=28.120,P01＜0.01;t0si=48.690,P0si＜0.01);与M0组比较,M2组巨噬细胞条件培养基中MFG-E8明显增高,M1及 si-MFG-E8 组明显降低(t02=17.240,P02＜0.01;t01=16.350,P01＜0.01;t0si=36.260,P0si＜0.01).CCK-8实验结果显示,与M0组比较,经M2型巨噬细胞条件培养基处理的HUVECs的增殖能力明显增加,M1 及 si-MFG-E8 组明显下降(t02=7.563,P02＜0.01;t01=4.365,P01＜0.01;t0si=5.965,P0si＜0.01).Transwell实验结果显示,与M0组比较,经M2型巨噬细胞条件培养基处理的HUVECs的迁移能力明显增加,M1及si-MFG-E8组明显下降,(t02=5.339,P02＜0.05;t01=2.991,P01＜0.05;t0si=6.275,P0si＜0.01).RT-PCR实验结果显示,与M0组比较,经M2型巨噬细胞条件培养基处理的HUVECs细胞PI3K、Akt和mTOR分子的表达水平明显升高,M1及si-MFG-E8组明显下降(PI3K:t02=5.393,Pg＜0.01;t01=4.35,P01＜0.05;t0si=4.718,P0si＜0.05;Akt:t02=4.849,P02＜0.05;t01=2.285,P01＞0.05;t0si=2.628,P0si＞0.05;mTOR:t02=6.128,P02＜0.01;t01=2.659,P01＞0.05;t0si=2.663,P0si＞0.05).结论 M2型巨噬细胞高表达MFG-E8,并可能通过上调PI3K、Akt和mTOR分子表达水平,促进脐静脉内皮细胞增殖和迁移.

The realization of coordinated infiltration of dermal and epidermal cell types, and the rapid deposition of extracellular matrix are the prerequisites for improved healing quality of large skin defects. However, presently used materials are limited for wound healing due to the inherent poor histocompatibility, low permeability, poor mechanical property and cytotoxicity. It is challenging to develop new therapeutic agents that enable timely and efficient wound healing and meanwhile there are minimal side effects to avoid chronic wounds, wound adhesions, and large scarring. Herein, we fabricated biomimetic exosomes (EMs) that enriched transforming growth factor β1 (TGF-β1) for the rapid healing of large-scale cutaneous wounds. The EM encapsulation allowed well-managed dosing of the TGF-β1 to effectively promote the wound healing process by endowing epidermal keratinocytes with a migratory feature, enhancing their stem cell properties, and resulting in accelerated in vivo wound re-epithelization. Specifically, the EMs could fast restore the function of sweat glands in a thermally injured mice model. Our work provides a promising strategy for improving the healing speed and healing quality for patients with large wounds.

Diabetic foot has a high incidence, long onset time and high treatment cost causing a significant economic burden on patients and their families. With the development of medicine, domestic and foreign experts pay more and more attention to the prevention of diabetic foot. This paper through the comprehensive analysis and induction of domestic and foreign literature, briefly expounds the risk factors of diabetic foot, discusses the importance of diabetic foot prevention, and introduces relevant measures for diabetic foot prevention, aiming to prevent the occurrence of diabetic foot, reduce the risk of diabetic foot complications, improve the quality of life, and provide reference for medical workers.

目的 利用3D生物打印技术将脱细胞真皮基质(ADM)及甲基丙烯酸酯化明胶(GelMA)混合制备成不同规格的皮肤复合支架,评价其生物相容性及对人脐静脉血管内皮细胞(HUVECs)生物学行为的影响.方法 将0％、0.75％、1.50％的ADM与GelMA混合配置成光敏性生物墨水,利用3D生物打印技术分别制备A组:单纯GelMA支架、B组:0.75％ADM/GelMA支架及C组:1.5％ADM/GelMA支架,扫描电镜观察其形态结构.通过CCK-8实验方法于第1、2、3天检测细胞增殖;将HUVECs与其共培养,通过Live/Dead染色观察皮肤支架表面细胞生长状态;使用Transwell小室实验检测皮肤支架对HUVECs迁移的影响;两组间比较采用非配对t检验,多组间比较采用单因素方差分析.结果 扫描电镜结果显示,各组皮肤复合支架均为十字交叉网格结构立体结构,且孔隙均匀,C组比A和B两组微观孔隙更加密集,孔隙之间的连通性更好.细胞计数试剂盒(CCK-8)实验结果显示,与阴性对照组(0 d:0.18±0.03;1 d:0.39±0.02;2 d:0.8±0.03;3 d:1.21±0.06)比较,在不同时间 A 组(0 d:0.19±0.04;1 d:0.39±0.03;2 d:0.75±0.03;3 d:1.19±0.06)细胞活力均无明显变化(F=1.37,P＞0.05),而 B 组(0d:0.21±0.05;1 d:0.6±0.04;2 d:1.15±0.04;3d:1.72±0.05)和C组(0d:0.22±0.06;1 d:0.59±0.03;2 d:1.18±0.05;3 d:1.87±0.02)细胞活力显著提高(F=22.85、26.76,P＜0.05).与A组比较,B组和C组在各时间点细胞活力显著提高(F=24.22、28.13,P＜0.05).Live/Dead实验染色结果显示,细胞在各组皮肤支架表面均能较好地黏附与生长,与A组(2.47±0.15)荧光强度比较,B组(4.63±0.35)和C组(11.98±0.31)荧光强度显著增高(t=10.00、48.26,P＜0.05),并且C组荧光强度明显高于B组(t=27.45,P＜0.05).Transwell小室实验结果显示,与A组(38.67±6.56)比较,B组(134±17.59)和C组(251.33±20.13)的血管内皮细胞迁移数显著增多(t=8.375、17.37,P＜0.05).结论 1.5％ADM/GelMA的皮肤支架具有较好的生物相容性和促进血管内皮细胞迁移的效应,有望为创面修复提供一种可降解的3D生物打印皮肤支架.

circRNAs have been considered as a rising factor in cancers. However, the roles and mechanisms of circ-sirt1 in gastric cancer (GC) remain largely unknown. In this study, we found that the expressions of sirt1 and circ-sirt1 are decreased in tissues or serums of GC patients by real-time quantitative PCR (RT-qPCR). The expressions of miR-132-3p/miR-212-3p showed an opposite tendency in these samples. The co-transfection of miR-132-3p/miR-212-3p mimics counteracted the enhancement of sirt1 expression induced by circ-sirt1. The results of cell colony-formation assay and transwell assays demonstrated that the proliferation, migration, and invasion activities of BGC-823 cells were inhibited by circ-sirt1 overexpression or miR-132-3p/miR-212-3p knockdown, respectively. The xenograft tumor model result indicated that the circ-sirt1 overexpression suppressed the tumor growth of BGC-823 cells. The regulation of miR-132-3p/miR-212-3p between circ-sirt1 and sirt1 was verified in the mice tumor tissues. Thus, circ-sirt1 inhibited tumor growth and invasion probably by sponging miR-132-3p/miR-212-3p and upregulating sirt1 expression in GC. These findings may provide a theoretical basis for the classification of GC and a novel therapeutic target for GC patients.

Objective:To investigate the effect of porcine acellular dermal matrix (pADM) on Cyclin D1 of rat keratinocytes and its mechanism.Methods:Primary rat keratinocytes were cultured in vitro and identified by immunohistochemistry. Following experimental groups were established: control group, pADM group, Wnt/β-catenin signal pathway agonist SKL2001 group, pADM+ Wnt/β-catenin signal pathway inhibitor FH535 group. The fluorescence quantitative PCR (qPCR) and the Western blotting were used to analyze the expression of Wnt/β-catenin signal pathway related molecules and Cyclin D1 in rat keratinocytes. Data were analyzed by Graph Pad Prism 8, and the measurement data were expressed by mean±standard deviation ( ± s). Unpaired t-test was used for comparison between the two groups, and one-way ANOVA was used for comparison between multiple groups. Results:The expression levels of Wnt/β-catenin signal pathway related molecules Wnt3a, β-catenin, LEF-1 and Cyclin D1 in pADM group were significantly higher than those in control group (for Wnt3a, qPCR: 2.489±0.414 vs. 1.210±0.242, t=4.068, P<0.05; Western blotting: 0.571±0.043 vs. 0.217±0.071, t=7.401, P<0.05; for β-catenin, qPCR: 2.594±0.244 vs. 0.818±0.188, t=9.877, P<0.05; Western blotting: 0.794±0.031 vs. 0.420±0.026, t=16.12, P<0.05); for LEF-1, qPCR: 2.816±0.190 vs. 0.896±0.090, t=9.702, P<0.05; Western blotting: 0.700±0.124 vs. 0.276±0.090, t=4.781, P<0.05; for Cyclin D1, qPCR: 2.708±0.305 vs. 0.922±0.095, t=15.720, P<0.05; Western blotting: 0.554±0.097 vs. 0.166±0.067, t=5.723, P<0.05). The expression levels of Wnt3a, β-catenin, LEF-1 and Cyclin D1 in pADM+ FH535 group were significantly lower than those in pADM group (for Wnt3a, qPCR: 0.350±0.049 vs. 0.964±0.159, F=49.890, P<0.05; Western blotting: 0.192±0.106 vs. 0.485±0.028, F=47.780, P<0.05; for β-catenin, qPCR: 0.431±0.149 vs. 0.919±0.074, F=39.430, P<0.05; Western blotting: 0.174±0.013 vs. 0.386±0.048, F=49.950, P<0.05; for LEF-1, qPCR: 0.502±0.067 vs. 0.954±0.112, F=79.130, P<0.05; Western blotting: 0.091±0.037 vs. 0.377±0.055, F=121.000, P<0.05; for Cyclin D1, qPCR: 0.264±0.049 vs. 0.962±0.037, F=105.500, P<0.05; Western blotting: 0.111±0.053 vs. 0.297±0.023, F=137.300, P<0.05). Conclusion:The effect of pADM is similar to that of Wnt/β-catenin signal pathway agonist, which can increase the expression of Cyclin D1 in rat keratinocytes by activating Wnt/β-catenin signal pathway, and FH535 can block the activation of Cyclin D1 by pADM.

Objective:To investigate whether vascular endothelial growth factor (VEGF) promotes the expression of vascular endothelial cell-related factor CD34 in keratinocytes.Methods:From October 2020 to July 2021, 15 C57BL/6 mice (purchased from Three Gorges University) were used to make full-thickness defect wounds on the back, and timely care was given. The normal tissues and the wound tissues on the 1, 3, 5 and 7 days were taken for Western blotting to detect the expression of VEGF. Cell experiments were divided into KCs group and KCs+ VEGF group. The level of CD34 messenger RNA (mRNA) was detected by real-time quantitative polymerase chain reaction (RT-qPCR). The protein expression levels of CD34, Delta-like ligand 4 (DLL4) and liver ligand B2 (Ephrin B2) were detected by Western blotting. The protein expression of CD34 was detected by Western blotting in VEGF group and VEGF+ VEGF inhibitor L-685458 group. The image processing software Image J and statistical software GraphPad Prism 6 were used for analysis. The t test was used for comparison between the two groups, and one-way ANOVA was used for multiple groups. Results:With the passage of time, the protein expression level of VEGF in the wound tissue increased dynamically (N=0.344±0.037, d1=0.158±0.019, d3=0.298±0.006, d5=0.509±0.066, d7=1.056±0.172, F=172.8, P<0.01). After VEGF was added, the mRNA expression of CD34 in keratinocytes increased (KCs=1.33±0.36, KCs+ VEGF=5.04±0.51, t=10.25, P<0.01), and the protein expression levels of CD34, DLL4 and Ephrin B2 increased (KCs=0.268±0.026, KCs+ VEGF=0.693±0.005, tCD34=27.78, PCD34<0.01; KCs=0.172±0.063, KCs+ VEGF=0.609±0.171, tDLL4=4.785, PDLL4<0.05; KCs=0.293±0.031, KCs+ VEGF=0.835±0.049, tEphrin B2=16.26, PEphrin B2<0.05). And after adding VEGF inhibitor, the expression of CD34 in keratinocytes was lower than that in VEGF group (VEGF=0.777±0.053, VEGF+ L-685458=0.263±0.053, t=11.83, P<0.01). Conclusion:VEGF promotes the expression of vascular endothelial cell-related factor CD34 in keratinocytes.

Objective:To investigate the effect of stromal cell-derived factor-1 (SDF-1) on the biological function of macrophages in high glucose environment and its mechanism.Methods:The Raw264.7 macrophage lines were established, and cell counting kit-8 (CCK-8) assay was used to detect the toxicity of SDF-1 on macrophages. Western blotting was used to detect the effects of high sugar (22.5 mmol/L) and normal sugar (5.5 mmol/L) on the expression of macrophane related protein. Following experimental groups were set up: group A (control), group B (SDF-1), group C (phosphatidyl inositol kinase/protein kinase B) inhibitor (LY294002), and group D (SDF-1+ LY294002). Transwell assay was used to examine the migration ability of giant macrophages. Western blotting was used to detect the changes in the expression of associated proteins. The comparison between the two groups was done by the non-paired t test, and the comparison among multiple groups was carried out by the multi-factor variance analysis. The P<0.05 was considered statistically significant. Results:The results of CCK-8 experiment showed that there was no significant difference among the groups ( P>0.05). Compared with normal glucose (5.5 mmol/L), high glucose (22.5 mmol/L) significantly decreased the expression of SDF-1, vascular endothelial growth factor (VEGF), SDF-1 specific receptor (CXCR4) and matrix metalloproteinase-9 (MMP-9) in macrophages ( t=50.510, 11.020, 27.660, 55.350, respectively, P<0.05). Transwell assay showed that the number of migrating macrophages in groups A, B, C and D was 86.630±4.096, 144.300±5.044, 59.000±2.646 and 65.670±4.256, respectively. Compared with group A, the t values of group B and group C were 21.460 and 5.574 respectively, and the P values were less than 0.05. In addition, SDF-1 could increase the expression of CXCR4, PI3K, Akt, MMP-9 and VEGF in macrophages, while LY294002 could reduce the expression of these proteins (for SDF-1 group, t=44.690, 5.393, 4.640, 13.250, 8.551, respectively, all P<0.05; for LY294002 group, t=13.430, 5.287, 4.381 and 45.510, respectively, all P<0.05). Conclusion:High glucose environment can affect the expression of SDF-1, VEGF, CXCR4 and MMP-9 in macrophages. SDF-1 may promote the secretion of MMP-9 and VEGF by macrophages by up-regulating the PI3K/Akt pathway.

Objective:To investigate the effect of porcine acellular dermal matrix (pADM) on the proliferation of hair follicle keratinocytes in vitro and its possible mechanism. Methods:Hair follicle keratinocytes were cultured in vitro and divided into 6 groups according to different intervention measures. group A, hair follicle keratinocytes only; group B, hair follicle keratinocytes treated with the pADM (20 mg/L); group C, hair follicle keratinocytes treated with tumor necrosis factor-α (TNF-α, 10 μg/L); group D, hair follicle keratinocytes treated with TNF-α inhibitor (Lenalidomide); group E, hair follicle keratinocytes treated with TNF-α (10 μg/L) and pADM (20 mg/L); group F, hair follicle keratinocytes treated with TNF-α inhibitor and pADM (20 mg/L). The real-time polymerase chain reaction (RT-PCR) method was used to detect the expression of LEF-1 mRNA and Ki-67 mRNA in each group. Immunofluorescence labeling method was used to detect hair follicle keratinocytes in groups B, E and F, and the effects of pADM on the proliferation of hair follicle keratinocytes were observed. Measurement data were expressed as mean±standard deviation ( mean± SD). The independent sample t test was used for comparison between the two groups, and P<0.05 indicated that the difference was statistically significant. Results:By RT-PCR analysis, the expressions of LEF-1 mRNA and Ki-67 mRNA in hair follicle keratinocytes of group A (LEF-1, 1.093±0.121, Ki-67, 1.100±0.089) were lower than those of group B (LEF-1, 2.973±0.234), Ki-67, 2.577±0.067) and group D (LEF-1, 1.867±0.045, Ki-67, 1.967±0.136), but higher than group C (LEF-1, 0.450±0.090, Ki-67, 0.490±0.026). The differences between group A and group B, group A and group C, group A and group D were statistically significant (A vs. B, tLEF-1=14.280, P<0.05; tKi-67=15.390, P<0.05; A vs. C, tLEF-1=4.887, P<0.05; tKi-67= 6.356, P<0.05; A vs. D, tLEF-1=5.875, P<0.05; tKi-67=9.030, P<0.05); Group B (LEF-1, 2.973±0.234, Ki-67, 2.577±0.067) The expression of LEF-1 mRNA and Ki-67 mRNA in hair follicle keratinocytes was higher than that of group E (LEF-1, 1.793±0.078, Ki-67, 1.940±0.165), but lower than group F (LEF-1, 4.257±0.266, Ki-67, 3.193±0.155), the differences between group B and group E, group B and group F were statistically significant (B vs. E, tLEF-1=8.964, P<0.05; tKi-67=6.634, P<0.05; B vs. F, tLEF-1=9.749, P<0.05; tKi-67=-6.425, P<0.05); immunofluorescence results showed that the number of hair follicle keratinocytes in group B was higher than that in group E but lower in Group F. Conclusion:In vitro, the pADM may up-regulate the expression of LEF-1 and Ki-67 by down-regulating TNF-α, and then promote the proliferation of hair follicle keratinocytes.