Characterization of γδ T cell subsets in organ transplantation

γδ T cells are innate‐type lymphocytes that preferentially act as regulators of local effector immune responses. Recent reports found an altered distribution of the two main subpopulations of blood γδ T cells (Vδ1 and Vδ2) in operationally tolerant liver transplant recipients. Based on this, γδ T cells subset quantification was proposed as a biomarker of immunologic risk in liver transplantation. The specific characteristics of γδ T cell subsets in transplantation remain however unknown. We have investigated here the phenotype, repertoire and functional properties of γδ T cell subsets in a large population of allograft recipients. Our results indicate that alterations in the γδ T cell compartment are not restricted to tolerant liver recipients. In fact, most immunosuppressed liver and kidney recipients also display an enlarged peripheral blood γδ T cell pool mainly resulting from an expansion of Vδ1 T cells exhibiting an oligoclonal repertoire and different phenotypic and cytokine production traits than Vδ2 T cells. We propose that persistent viral infections are likely to contribute to these alterations. Our data provide novel insight in the biology of γδ T cells and a rationale for exploring these lymphocytes in more depth into the pathogenesis of viral infections in transplantation.     

Reconstitution of T and NK cells after haploidentical hematopoietic cell transplantation using αβ T cell–depleted grafts and the clinical implication of γδ T cells

To investigate reconstitution of T and NK cells after αβ T lymphocyte–depleted haploidentical hematopoietic cell transplantation (HHCT) and the clinical implications of γδ T cells, we analyzed 50 pediatric patients who received 55 HHCTs using αβ T cell–depleted grafts. The number of CD3+ T cells and CD8+ T cells recovered rapidly and reached donor levels at days 180 and 60, respectively. Recovery of NK cells was rapid, and the median of NK cells at day 14 was comparable to the donor level. At day 14, median percentage of γδ T lymphocytes was 70.5%. After day 14, the percentage of γδ T cells gradually decreased, while the percentage of αβ T cells gradually increased. Patients with a low percentage (≤21%) of γδ T cells at day 30 had significantly higher incidence of cytomegalovirus (CMV) reactivation compared to patients with a high percentage (>70%) of γδ T cells (P < .01). In patients with acute leukemia, patients with high percentage of γδ T cells at day 30 showed significantly higher relapse‐free survival compared to those with low percentage of γδ T cells (P = .02). Data suggest that early recovery of γδ T cells decreases the risk of CMV reactivation and leukemia relapse.     

新型股骨髓内钉尾帽的研制与临床应用

目的研制一种用于封闭股骨交锁髓内钉中心孔尾端的新型尾帽，以减小尾帽植入髓内钉的难度，保证植入准确性，减少不必要的手术时间。方法2014 年成功研制出一种新型股骨髓内钉尾帽（以下简称“新型尾帽”）并获国家实用新型专利。采用临床随机对照研究，将 2014 年 8 月—2016 年 3 月收治的符合选择标准的 34 例非病理性股骨干骨折患者随机分为 A、B 两组，每组 17 例。两组患者性别、年龄、侧别、体质量指数、致伤原因、伤后至手术时间、骨折 AO 分型等一般资料比较差异均无统计学意义（P>0.05），具有可比性。两组患者均行交锁髓内钉固定术，A、B 组分别采用新型尾帽和原配尾帽。术中测量并比较两组植入尾帽的出血量和手术时间，术后比较两组并发症发生情况和骨折愈合时间，采用 Klemm 功能评定标准评价两组患肢功能。 结果A 组植入尾帽过程的出血量和手术时间分别为（3.5±0.8）mL 和（10.57±3.15）s，均显著少于 B 组的（5.5±1.7）mL 和（21.99±6.90）s，差异有统计学意义（t=–8.281，P=0.010；t=–10.743，P=0.009）。A 组新型尾帽均一次性植入成功，成功率 100%；B 组中 3 例原配尾帽旋入时脱落，一次性成功率 82.4%。两组患者均获随访，随访时间 6～16 个月，平均 9.7 个月。所有患者均无深部感染、内固定物松动或断裂以及其他严重并发症发生。A、B 组达骨折临床愈合时间分别为（16.4±3.2）周和（15.8±3.5）周，差异无统计学意义（t=0.514，P=0.611）。末次随访时，根据 Klemm 功能评定标准，A 组优 14 例、良 3 例，对照组优 11 例、良 6 例，差异无统计学意义（χ²=0.142，P=0.707）。 结论新型股骨髓内钉尾帽使术中操作简便，手术时间缩短，出血量减少，治疗效果满意。     

牛鼻子引流术在严重糖尿病足感染创面中的临床应用

目的探讨牛鼻子引流术（nose ring drain，NRD）治疗严重糖尿病足感染创面的临床效果。方法回顾分析 2017 年 6 月—2019 年 6 月采用 NRD 治疗的 35 例严重糖尿病足感染创面患者临床资料。男 24 例，女 11 例；年龄 28～82 岁，平均 54.5 岁。均为 2 型糖尿病患者，糖尿病病程 3～20 年，平均 9.4 年。糖尿病足病程 4 个月～2 年，平均 1.16 年。左足 16 例，右足 19 例。根据 Wagner 分级：2 级 11 例，3 级 20 例，4 级 4 例；均为糖尿病足创面中重度感染。术后创面采用“蚕食样”持续清创至表皮再生愈合。治疗过程中分析并总结创面分泌物细菌培养类型、临床抗生素使用时间，创面愈合方式、愈合时间、截肢比例等指标。结果35 例患者均随访，随访时间 3～6 个月，平均 4.5 个月。术后创面细菌培养结果显示，金黄色葡萄球菌 5 例、铜绿假单胞菌 4 例、大肠埃希菌 5 例、阴沟肠杆菌 3 例、凝固酶阴性葡萄球菌 3 例，其他类型 15 例。临床抗生素使用时间 3～15 d，平均 9.1 d。创面自溶性愈合，无需植皮，创面愈合时间 62～82 d，平均 72.3 d；随访过程中 3 例（8.6%）患者因血糖控制不佳，感染大面积扩散导致截肢 。创面愈合患者原创面处未出现感染复发及新生溃疡。结论应用 NRD 治疗严重糖尿病足感染创面，能有效控制创面感染，促进创面再生愈合，无需植皮且操作简便。     

Circulating γδ T cells and the risk of acute‐phase response after zoledronic acid administration

The use of intravenous nitrogen‐containing bisphosphonates (N‐BPs) is associated with the appearance of an acute phase response (APR) in a proportion of the patients for reasons that are poorly understood. The APR was attributed to the indirect activation of γδ T cells with the release of interferon‐γ and tumor necrosis factor (TNF). Forty patients with postmenopausal or senile osteoporosis (age range = 53–91 years) never previously treated with intravenous (iv) bisphosphonate, received a single 5‐mg zoledronic acid (ZOL) iv infusion over 15 minutes. White blood cells were counted and analyzed with an automated hematology analyzer (ADVIA 2120i Siemens, New York, USA) and by flow cytometer (BD FACSCanto, Becton Dickinson). The occurrence of APR was defined by the occurrence of fever (>37 °C) during the next 2 days. Forty‐two percent of patients (17 of 40) receiving the infusion of ZOL experienced an APR. Compared with the others they were younger (69 ± 7 years versus 74 ± 8 years; p = 0.06), and both the proportion and absolute number of γδ T cells were significant higher (p = 0.02 and p = 0.013, respectively). Nonsignificant differences were found between the two groups for white blood cells and for the other circulating lymphocyte subpopulations. Age was inversely correlated with circulating γδ T cells (p = 0.003) but the difference between the two groups in circulating γδ T cells persisted for age‐adjusted values and vice versa. In conclusion, the results of this study indicate that the number of circulating γδ T cells, together with age, are important determinant of the occurrence of APR after intravenous infusion of ZOL and possibly of any other N‐BPs. © 2012 American Society for Bone and Mineral Research     

选择性修薄股前外侧皮瓣修复足跟及足跟后区缺损

目的探讨选择性修薄股前外侧皮瓣修复足跟及足跟后区缺损的疗效。方法2013 年 4 月—2015 年 8 月，收治 8 例不同原因导致的足跟及足跟后区缺损患者。男 6 例，女 2 例；年龄 15～49 岁，平均 31.2 岁。创面范围为 14 cm×10 cm～19 cm×14 cm。手术切取对侧股前外侧皮瓣，将拟覆盖足底非承重区和足跟后区部分行不同程度修薄。皮瓣切取范围为 14 cm×10 cm～19 cm×14 cm。供区游离植皮修复。结果术后皮瓣均顺利成活，创面 Ⅰ 期愈合。2 例发生供区局部皮片坏死，经换药后愈合；其余植皮均顺利成活。患者均获随访，随访时间 8～20 个月，平均 12.3 个月。患者恢复正常行走，其中 2 例皮瓣发生破损，经换药后自愈。7 例皮瓣外观无臃肿，不影响穿鞋；1 例于术后 1 年行二期皮瓣修薄术。结论选择性修薄股前外侧皮瓣修复足跟及足跟后区缺损，可获得较理想足部外观和功能。     

SH3BP2 Cherubism Mutation Potentiates TNF‐α–Induced Osteoclastogenesis via NFATc1 and TNF‐α–Mediated Inflammatory Bone Loss

Cherubism (OMIM# 118400) is a genetic disorder with excessive jawbone resorption caused by mutations in SH3 domain binding protein 2 (SH3BP2), a signaling adaptor protein. Studies on the mouse model for cherubism carrying a P416R knock‐in (KI) mutation have revealed that mutant SH3BP2 enhances tumor necrosis factor (TNF)‐α production and receptor activator of nuclear factor‐κB ligand (RANKL)‐induced osteoclast differentiation in myeloid cells. TNF‐α is expressed in human cherubism lesions, which contain a large number of tartrate‐resistant acid phosphatase (TRAP)‐positive multinucleated cells, and TNF‐α plays a critical role in inflammatory bone destruction in homozygous cherubism mice (Sh3bp2^KI/KI). The data suggest a pathophysiological relationship between mutant SH3BP2 and TNF‐α–mediated bone loss by osteoclasts. Therefore, we investigated whether P416R mutant SH3BP2 is involved in TNF‐α–mediated osteoclast formation and bone loss. Here, we show that bone marrow–derived M‐CSF–dependent macrophages (BMMs) from the heterozygous cherubism mutant (Sh3bp2^KI/+) mice are highly responsive to TNF‐α and can differentiate into osteoclasts independently of RANKL in vitro by a mechanism that involves spleen tyrosine kinase (SYK) and phospholipase Cγ2 (PLCγ2) phosphorylation, leading to increased nuclear translocation of NFATc1. The heterozygous cherubism mutation exacerbates bone loss with increased osteoclast formation in a mouse calvarial TNF‐α injection model as well as in a human TNF‐α transgenic mouse model (hTNFtg). SH3BP2 knockdown in RAW264.7 cells results in decreased TRAP‐positive multinucleated cell formation. These findings suggest that the SH3BP2 cherubism mutation can cause jawbone destruction by promoting osteoclast formation in response to TNF‐α expressed in cherubism lesions and that SH3BP2 is a key regulator for TNF‐α–induced osteoclastogenesis. Inhibition of SH3BP2 expression in osteoclast progenitors could be a potential strategy for the treatment of bone loss in cherubism as well as in other inflammatory bone disorders. © 2014 American Society for Bone and Mineral Research.     

多种皮瓣联合修复面部痣样基底细胞癌综合征术后缺损一例

目的总结1例多种皮瓣联合修复面部痣样基底细胞癌综合征(nevoid basal cell carcinoma syndrome,NBCCS)术后缺损疗效。方法2019年6月收治1例29岁NBCCS女性患者,因面部多发黑色丘疹10余年入院。入院检查,面部可见多发黑色斑块(范围为0.6 cm×0.3 cm^3.5 cm×3.0 cm)以及散在黑褐色丘疹(直径0.2~0.6 cm)。术前病理检查示BCC。术中行肿物扩大切除后软组织缺损范围为0.6 cm×0.4 cm^7.5 cm×3.5 cm,分别采用鼻唇沟瓣、局部皮瓣及前臂皮瓣修复。结果术后各皮瓣均成活,切口Ⅰ期愈合。患者获随访5个月,面部表情自然,外形对称,右眶下区与周围组织存在一定色差。患者及家属对手术效果满意。结论面部NBCCS术后缺损可选择多种皮瓣联合修复,其中前臂皮瓣可修复面部较大缺损,鼻唇沟瓣、局部皮瓣修复小缺损。     

Indirubin‐3′‐Oxime Reverses Bone Loss in Ovariectomized and Hindlimb‐Unloaded Mice Via Activation of the Wnt/β‐Catenin Signaling

Osteoporosis is a major global health issue in elderly people. Because Wnt/β‐catenin signaling plays a key role in bone homeostasis, we screened activators of this pathway through cell‐based screening, and investigated indirubin‐3′‐oxime (I3O), one of the positive compounds known to inhibit GSK3β, as a potential anti‐osteoporotic agent. Here, we show that I3O activated Wnt/β‐catenin signaling via inhibition of the interaction of GSK3β with β‐catenin, and induced osteoblast differentiation in vitro and increased calvarial bone thickness ex vivo. Intraperitoneal injection of I3O increased bone mass and improved microarchitecture in normal mice and reversed bone loss in an ovariectomized mouse model of age‐related osteoporosis. I3O also increased thickness and area of cortical bone, indicating improved bone strength. Enhanced bone mass and strength correlated with activated Wnt/β‐catenin signaling, as shown by histological analyses of both trabecular and cortical bones. I3O also restored mass and density of bone in hindlimb‐unloaded mice compared with control, suspended mice, demonstrating bone‐restoration effects of I3O in non‐aged–related osteoporosis as well. Overall, I3O, a pharmacologically active small molecule, could be a potential therapeutic agent for the treatment and prevention of osteoporosis. © 2014 American Society for Bone and Mineral Research.     

手术时机对胸椎结核伴脊髓损害疗效的影响

目的探讨后路病灶清除脊髓减压植骨内固定治疗伴脊髓损害的胸椎结核的可行性，比较手术时机对术后疗效的影响。方法回顾分析 2012 年 8 月—2015 年 10 月收治的 26 例胸椎结核伴脊髓损害患者临床资料。所有患者行后方经单侧椎弓根入路病灶清除脊髓减压植骨内固定术，其中 11 例出现脊髓损害症状至手术时间<3 个月（A 组）、15 例出现脊髓损害症状至手术时间>3 个月（B 组）。两组患者性别、年龄、病变节段及术前红细胞沉降率、C 反应蛋白、病变节段 Cobb 角、美国脊髓损伤协会（ASIA）分级等一般资料比较，差异均无统计学意义（P>0.05），具有可比性。记录并比较两组患者手术时间、术中出血量、住院时间、围术期并发症发生情况及骨融合时间；比较两组术前及末次随访时 Cobb 角并计算矫正度；采用 ASIA 分级评价手术前后脊髓功能。 结果两组患者均获随访，随访时间 25～60 个月，平均 41.6 个月。术中、术后均无脑脊液漏发生。A 组住院时间及围术期并发症均显著少于 B 组（P<0.05）；两组手术时间、术中出血量及骨融合时间比较差异均无统计学意义（P>0.05）。末次随访时 A、B 组红细胞沉降率和 C 反应蛋白比较差异无统计学意义（P>0.05），但均较术前明显降低（P<0.05）。A 组 1 例 T_6、7 结核术后切口出现窦道，经换药等处理后愈合，20 个月骨性融合后取出内固定物，36 个月随访时结核无复发；B 组 1 例 T_4、5 结核术后 26 个月时复发并出现胸椎畸形，予以翻修。其余患者均无内固定物相关并发症或结核复发。末次随访时两组患者病变节段 Cobb 角均较术前显著改善（P<0.05），两组间 Cobb 角及矫正度比较差异均无统计学意义（P>0.05）。末次随访时脊髓功能 ASIA 分级，A 组 C 级 1 例、E 级 10 例，B 组 D 级 2 例、E 级 13 例，均较术前显著改善（P<0.05）；两组间比较差异无统计学意义（Z=–0.234，P=1.000）。 结论对伴有脊髓损害的胸椎结核患者，采用后方经单侧椎弓根入路病灶清除脊髓减压植骨内固定术治疗可以取得满意效果，尽早手术可以缩短患者住院时间、降低围术期并发症发生风险。     

改良诱导膜技术结合带蒂（肌）皮瓣治疗糖尿病患者胫骨慢性骨髓炎

目的探讨改良诱导膜技术结合带蒂（肌）皮瓣治疗糖尿病患者胫骨慢性骨髓炎的疗效。方法回顾分析 2017 年 1 月—2019 年 3 月，收治并符合选择标准的 22 例合并胫骨慢性骨髓炎的糖尿病患者临床资料。男 15 例，女 7 例；年龄 44～65 岁，平均 52 岁。糖尿病病程 3～12 年，平均 6.1 年。骨髓炎病程 4 个月～7 年，平均 3.3 年；Cierny-Mader 分型：Ⅲ型 9 例，Ⅳ型 13 例。细菌培养显示单一细菌感染 21 例，混合细菌感染 1 例。术前下肢动脉彩色多普勒超声及 CT 血管造影检查证实胫前、胫后动脉通畅无闭塞。第 1 阶段治疗首先彻底清创，病灶清除后骨缺损长 4～9 cm，平均 5.6 cm；软组织缺损范围 5 cm×2 cm～10 cm×7 cm。采用抗生素骨水泥分别填塞骨与软组织缺损区域；7～10 d 后取出骨水泥填塞物，另取抗生素骨水泥填塞骨缺损区域，同时行带蒂（肌）皮瓣修复创面。待 7～12 周炎症指标均恢复正常时行第 2 阶段治疗，即采用自体髂骨或联合人工骨植骨修复缺损。观察创面及骨缺损愈合情况以及并发症发生情况，记录治疗成功例数，评价皮瓣疗效满意度及患肢功能。结果术后 3 例发生皮瓣边缘局部表皮坏死，创面延期愈合；其余 19 例皮瓣均顺利成活，创面Ⅰ期愈合。供区植皮均成活，切口Ⅰ期愈合。患者均获随访，随访时间 13～28 个月，平均 20 个月。术后 12 个月内 2 例感染复发，再次采用改良诱导膜技术治疗后骨缺损愈合；骨缺损愈合率达 100%，愈合时间 6～10 个月，平均 8.9 个月；感染控制率和治疗成功率均为 90.9%（20/22）。术后 12 个月，参照张浩等制定的皮瓣疗效满意度评分标准疗效均为满意；患肢功能按 Johner-Wruhs 邻近关节功能评定方法获优 13 例、良 7 例、可 2 例，优良率为 90.9%。结论无血管闭塞的糖尿病患者发生胫骨慢性骨髓炎时，采用改良诱导膜技术结合带蒂（肌）皮瓣能修复骨与软组织缺损同时控制感染，早中期疗效良好。     

Proteomic identification of 14‐3‐3ϵ as a linker protein between pERK1/2 inhibition and BIM upregulation in human osteosarcoma cells

Despite advancements in multimodality chemotherapy, conventional cytotoxic treatments still remain ineffective for a subset of patients with aggressive metastatic or multifocal osteosarcoma. It has been shown that pERK1/2 inhibition enhances chemosensitivity to doxorubicin and promotes osteosarcoma cell death in vivo and in vitro. One of the pro‐apoptotic mechanisms is upregulation of Bim by pERK1/2 inhibitors. To this end, we examined proteomic changes of 143B human osteosarcoma cells with and without treatment of PD98059, pERK1/2 inhibitor. Specifically, we identified 14‐3‐3? protein as a potential mediator of Bim expression in response to inhibition of pERK1/2. We hypothesized that 14‐3‐3? mediates upregulation of Bim expression after pERK1/2 inhibition. We examined the expression of Bim after silencing 14‐3‐3? using siRNA. The 14‐3‐3? gene silencing resulted in downregulation of Bim expression after PD98059 treatment. These data indicate that 14‐3‐3? is required for Bim expression and that it has an anti‐cancer effect under pERK1/2 inhibition in 143B cells. By playing an essential role upstream of Bim, 14‐3‐3? may potentially be a coadjuvant factor synergizing the effect of pERK1/2 inhibitors in addition to conventional cytotoxic agents for more effective osteosarcoma treatments. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:848–854, 2014.

     

接力皮瓣修复耳廓及供瓣区皮肤软组织缺损

目的探讨接力皮瓣修复耳廓及供瓣区皮肤软组织缺损的疗效。方法2014 年 5 月—2016 年 1 月，收治 10 例耳廓皮肤肿瘤患者。男 4 例，女 6 例；年龄 21～69 岁，平均 35 岁。基底细胞癌 2 例，色素痣 3 例，乳头状瘤 4 例，鲍文病 1 例。术中切除肿物后遗留创面范围为 1.1 cm×1.0 cm～2.3 cm×1.7 cm。用颞浅动脉耳后穿支皮瓣修复耳廓缺损，皮瓣切取范围为 1.5 cm×1.4 cm～2.8 cm×2.0 cm；使用耳后动脉穿支顺行推进皮瓣修复第 1 供瓣区，皮瓣切取范围为 4.0 cm×2.0 cm～7.5 cm×3.0 cm，第 2 供瓣区创面直接拉拢缝合。结果术后皮瓣均顺利成活，供、受区创面均Ⅰ期愈合。患者均获随访，随访时间 10～28 个月，平均 14.2 个月。重建耳廓形态满意，肿瘤无复发。皮瓣外形、色泽、质地和厚度与受区基本一致，瘢痕不明显，无牵拉畸形，感觉无明显异常。结论接力皮瓣血供可靠，手术操作简便，供区隐蔽，是修复耳廓及供瓣区皮肤软组织缺损的较好选择。     

足背动脉穿支蒂皮瓣修复前足背侧皮肤软组织缺损疗效观察

目的探讨足背动脉穿支蒂皮瓣修复前足背侧皮肤软组织缺损的临床疗效。方法2011 年 1 月—2016 年 6 月，应用足背动脉穿支蒂皮瓣修复前足背侧皮肤软组织缺损 16 例。男 10 例，女 6 例；年龄 17～65 岁，平均 28.5 岁。致伤原因：重物砸伤 9 例，交通事故伤 7 例。左足 7 例，右足 9 例。创面均位于跖跗关节以远前足背侧；合并肌腱外露 10 例，骨外露 6 例。急症修复 9 例，择期修复 7 例。创面范围 5.0 cm×4.0 cm～7.5 cm×7.0 cm。皮瓣切取范围为 6 cm×6 cm～9 cm×9 cm。结果1 例皮瓣术后第 2 天开始出现张力性水疱，皮瓣肿胀较严重，经对症处理后愈合；其余皮瓣及供区植皮均成活。术后 16 例均获随访，随访时间 6～18 个月，平均 8 个月。皮瓣颜色、质地、厚度均与周围正常组织相似，患足能正常穿鞋及负重行走。结论足背动脉穿支蒂皮瓣手术操作简便，无需牺牲主干血管，是修复前足背侧皮肤软组织缺损的有效方法之一。