Activatable near-infrared fluorescence probe for real-time imaging of PD-L1 expression in tumors.

In clinical practice, determining programmed death-ligand 1 (PD-L1) expression is crucial for selecting patients and monitoring immune checkpoint blockade therapies. Currently, PD-L1 expression is quantified using immunohistochemistry (IHC). However, IHC-based methods do not capture the heterogeneous and dynamic nature of PD-L1 expression. Thus, there is a pressing need for a rapid and efficient method for monitoring PD-L1 expression both in vitro and in vivo, which would considerably aid in prognosis and treatment selection. In this study, we present for the first time an activatable near-infrared (NIR) fluorescence imaging probe (Q-Atezol) for the real-time monitoring of PD-L1 expression in vitro and in vivo. The ability of Q-Atezol to detect PD-L1 expression quickly and in real-time was evaluated in both tumor spheroid and lung cancer xenograft models. An always-on optical probe (ON-Atezol) was synthesized and tested for comparison. In vivo NIR fluorescence imaging studies were conducted on A549 and H1975 tumor-bearing mice, and their tumor-to-background ratios (TBRs) were analyzed. The quenched NIR fluorescence of Q-Atezol is activated upon binding to PD-L1 proteins on the surface of cancer cells, thereby enabling PD-L1 detection in the three-dimensional (3D) tumor spheroids without a washing step. Notably, PD-L1-positive H1975 tumors were clearly visualized with a high TBR 6 hours after Q-Atezol injection, whereas ON-Atezol treatment could not detect H1975 tumors even 24 hours post-injection. The activatable fluorescence probe Q-Atezol demonstrated great potential as an exceptional sensor for assessing PD-L1 expression in 3D cell structures and for in vivo applications.

Quantitative Assessment of Condylar Remodeling After Open Reduction and Internal Fixation in Mandibular Condylar Head Fractures.

Open reduction and internal fixation (OR/IF) and closed treatments are viable options, with OR/IF gaining popularity in mandibular condylar head fracture (CHF). Sufficient research is lacking on long-term condylar morphologic changes. This study quantitatively evaluated the morphologic changes in the condylar head in patients who underwent OR/IF for CHF without the removal of osteosynthesis material. A retrospective study analyzed patients who underwent OR/IF for CHF between November 2010 and August 2022. The transverse/lateral condyle width and condyle height were radiologically analyzed using panoramic and reverse Towne view x-ray images at immediate (T0), short-term (T1), and long-term (T2) postoperative periods. The study involved 29 patients with 32 surgically treated CHF. Long-term condyle height decreased significantly from 18.9±0.8 mm at T0 to 18.1±0.8 mm at T1 (P=0.042), and further to 17.3±0.7 mm at T2 (P=0.034). Transverse width slightly reduced from 18.7±0.7 mm at T0 to 17.9±0.8 mm at T1 (P=0.001) but remained stable from T1 to T2 (17.6±1.0 mm, P=0.756). Following surgery, maximal mouth opening averaged 39.5±6.1 mm, with one case of chin deviation and three cases of joint pain. The condyle undergoes long-term shortening without altering its width. Nevertheless, patients experience favorable functional outcomes, including satisfactory mouth opening and occlusion. In addition, it is advisable not to remove the osteosynthesis material, as the remaining material causes minimal functional impairment and bone resorption.

PolyGR and polyPR knock-in mice reveal a conserved neuroprotective extracellular matrix signature in C9orf72 ALS/FTD neurons.

Dipeptide repeat proteins are a major pathogenic feature of C9orf72 amyotrophic lateral sclerosis (C9ALS)/frontotemporal dementia (FTD) pathology, but their physiological impact has yet to be fully determined. Here we generated C9orf72 dipeptide repeat knock-in mouse models characterized by expression of 400 codon-optimized polyGR or polyPR repeats, and heterozygous C9orf72 reduction. (GR)400 and (PR)400 knock-in mice recapitulate key features of C9ALS/FTD, including cortical neuronal hyperexcitability, age-dependent spinal motor neuron loss and progressive motor dysfunction. Quantitative proteomics revealed an increase in extracellular matrix (ECM) proteins in (GR)400 and (PR)400 spinal cord, with the collagen COL6A1 the most increased protein. TGF-ß1 was one of the top predicted regulators of this ECM signature and polyGR expression in human induced pluripotent stem cell neurons was sufficient to induce TGF-ß1 followed by COL6A1. Knockdown of TGF-ß1 or COL6A1 orthologues in polyGR model Drosophila exacerbated neurodegeneration, while expression of TGF-ß1 or COL6A1 in induced pluripotent stem cell-derived motor neurons of patients with C9ALS/FTD protected against glutamate-induced cell death. Altogether, our findings reveal a neuroprotective and conserved ECM signature in C9ALS/FTD.

An Analysis of the Demographics and Clinical Characteristics of Transgender and Intersex Populations in Korea: A Retrospective Study Using HIRA Database.

BACKGROUND: Transgender and intersex populations have long remained under-documented in South Korea, largely due to the absence of comprehensive epidemiological data. With increasing societal acknowledgment, there's an urgent need to understand the demographics and health challenges faced by these communities. METHODS: This retrospective, large-scale data study included people who received the F64 codes from the Korean Health Insurance Review & Assessment Service between January 2007 and December 2021. Demographics, gender-affirmative treatments, and psychiatric related medications were examined. RESULTS: Between 2007 and 2021, 8,602 patients were diagnosed with "gender identity disorder" and 45 with "intersex." A steadily increasing annual prevalence was observed, peaking at 986 cases in 2021. The majority (79.8%) were aged between 10 and 30. Nearly half (53.2%) exhibited mental and behavioral disorders. Two-thirds had been prescribed anxiolytics or sedatives either before or after diagnosis. Merely 12.1% received hormone therapy covered by health insurance. CONCLUSION: This is the first large-scale study highlighting the demographics and clinical characteristics of the transgender and intersex populations in Korea. The study reveals a consistent growth of these communities over the past 15 years, with a significant proportion under 30 years of age facing mental and behavioral challenges. Findings underscore the need for targeted healthcare interventions, early psychological support, and comprehensive insurance coverage tailored to the specific needs of these individuals in Korea.

Crystal structure of [1,2,4]triazolo[4,3-b]pyridazine derivatives as BRD4 bromodomain inhibitors and structure-activity relationship study.

BRD4 contains two tandem bromodomains (BD1 and BD2) that recognize acetylated lysine for epigenetic reading, and these bromodomains are promising therapeutic targets for treating various diseases, including cancers. BRD4 is a well-studied target, and many chemical scaffolds for inhibitors have been developed. Research on the development of BRD4 inhibitors against various diseases is actively being conducted. Herein, we propose a series of [1,2,4]triazolo[4,3-b]pyridazine derivatives as bromodomain inhibitors with micromolar IC50 values. We characterized the binding modes by determining the crystal structures of BD1 in complex with four selected inhibitors. Compounds containing [1,2,4] triazolo[4,3-b]pyridazine derivatives offer promising starting molecules for designing potent BRD4 BD inhibitors.

BMS794833 inhibits macrophage efferocytosis by directly binding to MERTK and inhibiting its activity.

Myeloid epithelial reproductive proto-oncogene tyrosine kinase (MERTK) plays an essential role in modulating cancer immune tolerance by regulating macrophage efferocytosis. Studies are underway to develop small-molecule chemicals that inhibit MERTK as cancer immunotherapeutic agents, but these efforts are in their early stages. This study identified BMS794833, whose primary targets are MET and VEGFR2, as a potent MERTK inhibitor and developed a real-time efferocytosis monitoring system. The X-ray cocrystal structure revealed that BMS794833 was in contact with the ATP-binding pocket and the allosteric back pocket, rendering MERTK inactive. Homogeneous time-resolved fluorescence kinetic and Western blotting analyses showed that BMS794833 competitively inhibited MERTK activity in vitro and inhibited the autophosphorylation of MERTK in macrophages. We developed a system to monitor MERTK-dependent efferocytosis in real time, and using this system, we confirmed that BMS794833 significantly inhibited the efferocytosis of differentiated macrophages. Finally, BMS794833 significantly inhibited efferocytosis in vivo in a mouse model. These data show that BMS794833 is a type II MERTK inhibitor that regulates macrophage efferocytosis. In addition, the real-time efferocytosis monitoring technology developed in this study has great potential for future applications.

The structure of Deinococcus radiodurans transcriptional regulator HucR retold with the urate bound.

HucR is a MarR family protein of Deinococcus radiodurans, which binds tightly to the intergenic region of HucR and the uricase gene to inhibit their expression. Urate (or uric acid) antagonizes the repressor function of HucR by binding to HucR to impede its association with the cognate DNA. The previously reported crystal structure of HucR was without the bound urate showing significant structural homology to other MarR structures. In this paper, we report the crystal structure of HucR determined with the urate bound. However, despite the fact that the urate is found at a site well-known to harbor ligands in other MarR family proteins, the overall HucR structure indicates that no significant change in structure takes place with the urate bound. Structure analysis further suggests that the urate interaction in HucR is mediated by histidine/glutamate side chains and ordered water molecules stabilized by various residues. Such interaction is quite unique compared to other known structural interactions between urate and its binding proteins. Furthermore, structural comparison of the apo- and the urate bound forms allows us to hypothesize that the Trp20-mediated water network in the apo-form stabilizes the proper HucR fold for cognate DNA binding, and that urate binding, also via Trp20, and the consequent reorganization of water molecules in the binding pocket, likely disrupts the DNA binding configuration to result in the attenuated DNA binding.

Osteogenic Capacity of the Prefabricated Periosteofascial Flap using Vascular Induction with Skeletonized Pedicle Transfer in Rabbit Calvarium.

The study investigated the osteogenic capacity of a prefabricated periosteal flap created using only skeletonized pedicle transfer without fascia or muscle for vascular induction in rabbit calvarium. A critical-sized bone defect was made in the parietal bone centered on the sagittal suture, and the demineralized bone matrix was implanted. The periosteofascia over the defect was used as a form of prefabricated periosteofascial flap (PPF group, N=10), conventional periosteofascial flap (CPF group, N=10), and nonvascularized free periosteofascial graft (FPG group, N=6). The prefabricated flap was designed via vascular induction by transferring the central artery and vein of the right auricle onto the periosteofascia for 4 weeks prior to flap elevation. A quantitative comparison of volume restoration and radiodensity in the bone defect and a histological study were performed after 6 weeks of covering the bone defect with periosteofascia. The volume restoration of the bone defect covered with the PPF (43.4%) was not different from that of the CPF (46.2%), but significantly increased compared with that of the FPG (24.6%). The radiodensity of the bone defect covered with the PPF (-186.3 HU) was not different from that of the CPF (-153.6 HU), but significantly increased compared with that of the FPG (-329.8 HU). The results were based on adequate vascular development of the periosteum and were closely related to the osteogenic changes in the implanted demineralized bone matrix (DBM). In conclusion, even in the PPF created by transferring only skeletonized vascular pedicles, the osteogenic capacity of the periosteofascial flap is well maintained.

The Reciprocal Interaction Between Sleep and Alzheimer's Disease.

It is becoming increasingly recognized that patients with a variety of neurodegenerative diseases exhibit disordered sleep/wake patterns. While sleep impairments have typically been thought of as sequelae of underlying neurodegenerative processes in sleep-wake cycle regulating brain regions, including the brainstem, hypothalamus, and basal forebrain, emerging evidence now indicates that sleep deficits may also act as pathophysiological drivers of brain-wide disease progression. Specifically, recent work has indicated that impaired sleep can impact on neuronal activity, brain clearance mechanisms, pathological build-up of proteins, and inflammation. Altered sleep patterns may therefore be novel (potentially reversible) dynamic functional markers of proteinopathies and modifiable targets for early therapeutic intervention using non-invasive stimulation and behavioral techniques. Here we highlight research describing a potentially reciprocal interaction between impaired sleep and circadian patterns and the accumulation of pathological signs and features in Alzheimer's disease, the most prevalent neurodegenerative disease in the elderly.

Phosphorylation of OGFOD1 by Cell Cycle-Dependent Kinase 7/9 Enhances the Transcriptional Activity of RNA Polymerase II in Breast Cancer Cells.

2-oxoglutarate and iron-dependent oxygenase domain-containing protein 1 (OGFOD1) expression is upregulated in a variety of cancers and has been related to poor prognosis. However, despite this significance to cancer progression, the precise oncogenic mechanism of OGFOD1 is not understood. We demonstrated that OGFOD1 plays a role in enhancing the transcriptional activity of RNA polymerase II in breast cancer cells. OGFOD1 directly binds to the C-terminal domain of RNA polymerase II to alter phosphorylation status. The elimination of OGFOD1 resulted in decreased tumor development. Additionally, cell cycle-dependent kinase 7 and cell cycle-dependent kinase 9, critical enzymes for activating RNA polymerase II, phosphorylated serine 256 of OGFOD1, whereas a non-phosphorylated mutant OGFOD1 failed to enhance transcriptional activation and tumor growth. Consequently, OGFOD1 helps promote tumor growth by enhancing RNA polymerase II, whereas simultaneous phosphorylation of OGFOD1 by CDK enzymes is essential in stimulating RNA polymerase II-mediated transcription both in vitro and in vivo, and expression of target genes.

Phospholipid transfer function of PTPIP51 at mitochondria-associated ER membranes.

In eukaryotic cells, mitochondria are closely tethered to the endoplasmic reticulum (ER) at sites called mitochondria-associated ER membranes (MAMs). Ca2+ ion and phospholipid transfer occurs at MAMs to support diverse cellular functions. Unlike those in yeast, the protein complexes involved in phospholipid transfer at MAMs in humans have not been identified. Here, we determine the crystal structure of the tetratricopeptide repeat domain of PTPIP51 (PTPIP51_TPR), a mitochondrial protein that interacts with the ER-anchored VAPB protein at MAMs. The structure of PTPIP51_TPR shows an archetypal TPR fold, and an electron density map corresponding to an unidentified lipid-like molecule probably derived from the protein expression host is found in the structure. We reveal functions of PTPIP51 in phospholipid binding/transfer, particularly of phosphatidic acid, in vitro. Depletion of PTPIP51 in cells reduces the mitochondrial cardiolipin level. Additionally, we confirm that the PTPIP51-VAPB interaction is mediated by the FFAT-like motif of PTPIP51 and the MSP domain of VAPB. Our findings suggest that PTPIP51 is a phospholipid transfer protein with a MAM-tethering function.

Synthesis and Structure-Activity Relationships of Aristoyagonine Derivatives as Brd4 Bromodomain Inhibitors with X-ray Co-Crystal Research.

Epigenetic regulation is known to play a key role in progression of anti-cancer therapeutics. Lysine acetylation is an important mechanism in controlling gene expression. There has been increasing interest in bromodomain owing to its ability to modulate transcription of various genes as an epigenetic 'reader.' Herein, we report the design, synthesis, and X-ray studies of novel aristoyagonine (benzo[6,7]oxepino[4,3,2-cd]isoindol-2(1H)-one) derivatives and investigate their inhibitory effect against Brd4 bromodomain. Five compounds 8ab, 8bc, 8bd, 8be, and 8bf have been discovered with high binding affinity over the Brd4 protein. Co-crystal structures of these five inhibitors with human Brd4 bromodomain demonstrated that it has a key binding mode occupying the hydrophobic pocket, which is known to be the acetylated lysine binding site. These novel Brd4 bromodomain inhibitors demonstrated impressive inhibitory activity and mode of action for the treatment of cancer diseases.

Crystal Structure of the Kinase Domain of MerTK in Complex with AZD7762 Provides Clues for Structure-Based Drug Development.

Aberrant tyrosine-protein kinase Mer (MerTK) expression triggers prosurvival signaling and contributes to cell survival, invasive motility, and chemoresistance in many kinds of cancers. In addition, recent reports suggested that MerTK could be a primary target for abnormal platelet aggregation. Consequently, MerTK inhibitors may promote cancer cell death, sensitize cells to chemotherapy, and act as new antiplatelet agents. We screened an inhouse chemical library to discover novel small-molecule MerTK inhibitors, and identified AZD7762, which is known as a checkpoint-kinase (Chk) inhibitor. The inhibition of MerTK by AZD7762 was validated using an in vitro homogeneous time-resolved fluorescence (HTRF) assay and through monitoring the decrease in phosphorylated MerTK in two lung cancer cell lines. We also determined the crystal structure of the MerTK:AZD7762 complex and revealed the binding mode of AZD7762 to MerTK. Structural information from the MerTK:AZD7762 complex and its comparison with other MerTK:inhibitor structures gave us new insights for optimizing the development of inhibitors targeting MerTK.

Evaluation of the clinical efficacy of a TW3-based fully automated bone age assessment system using deep neural networks.

PURPOSE: The aim of this study was to evaluate the clinical efficacy of a Tanner-Whitehouse 3 (TW3)-based fully automated bone age assessment system on hand-wrist radiographs of Korean children and adolescents. MATERIALS AND METHODS: Hand-wrist radiographs of 80 subjects (40 boys and 40 girls, 7-15 years of age) were collected. The clinical efficacy was evaluated by comparing the bone ages that were determined using the system with those from the reference standard produced by 2 oral and maxillofacial radiologists. Comparisons were conducted using the paired t-test and simple regression analysis. RESULTS: The bone ages estimated with this bone age assessment system were not significantly different from those obtained with the reference standard (P>0.05) and satisfied the equivalence criterion of 0.6 years within the 95% confidence interval (- 0.07 to 0.22), demonstrating excellent performance of the system. Similarly, in the comparisons of gender subgroups, no significant difference in bone age between the values produced by the system and the reference standard was observed (P>0.05 for both boys and girls). The determination coefficients obtained via regression analysis were 0.962, 0.945, and 0.952 for boys, girls, and overall, respectively (P=0.000); hence, the radiologist-determined bone ages and the system-determined bone ages were strongly correlated. CONCLUSION: This TW3-based system can be effectively used for bone age assessment based on hand-wrist radiographs of Korean children and adolescents.

Phosphorylation of OCT4 Serine 236 Inhibits Germ Cell Tumor Growth by Inducing Differentiation.

Octamer-binding transcription factor 4 (Oct4) plays an important role in maintaining pluripotency in embryonic stem cells and is closely related to the malignancies of various cancers. Although posttranslational modifications of Oct4 have been widely studied, most of these have not yet been fully characterized, especially in cancer. In this study, we investigated the role of phosphorylation of serine 236 of OCT4 [OCT4 (S236)] in human germ cell tumors (GCTs). OCT4 was phosphorylated at S236 in a cell cycle-dependent manner in a patient sample and GCT cell lines. The substitution of endogenous OCT4 by a mimic of phosphorylated OCT4 with a serine-to-aspartate mutation at S236 (S236D) resulted in tumor cell differentiation, growth retardation, and inhibition of tumor sphere formation. GCT cells expressing OCT4 S236D instead of endogenous OCT4 were similar to cells with OCT4 depletion at the mRNA transcript level as well as in the phenotype. OCT4 S236D also induced tumor cell differentiation and growth retardation in mouse xenograft experiments. Inhibition of protein phosphatase 1 by chemicals or short hairpin RNAs increased phosphorylation at OCT4 (S236) and resulted in the differentiation of GCTs. These results reveal the role of OCT4 (S236) phosphorylation in GCTs and suggest a new strategy for suppressing OCT4 in cancer.

Volumetric lung nodule segmentation using adaptive ROI with multi-view residual learning.

Accurate quantification of pulmonary nodules can greatly assist the early diagnosis of lung cancer, enhancing patient survival possibilities. A number of nodule segmentation techniques, which either rely on a radiologist-provided 3-D volume of interest (VOI) or use the constant region of interests (ROIs) for all the slices, are proposed; however, these techniques can only investigate the presence of nodule voxels within the given VOI. Such approaches restrain the solutions to freely investigate the nodule presence outside the given VOI and also include the redundant structures (non-nodule) into VOI, which limits the segmentation accuracy. In this work, a novel semi-automated approach for 3-D segmentation of lung nodule in computerized tomography scans, has been proposed. The technique is segregated into two stages. In the first stage, a 2-D ROI containing the nodule is provided as an input to perform a patch-wise exploration along the axial axis using a novel adaptive ROI algorithm. This strategy enables the dynamic selection of the ROI in the surrounding slices to investigate the presence of nodules using a Deep Residual U-Net architecture. This stage provides the initial estimation of the nodule utilized to extract the VOI. In the second stage, the extracted VOI is further explored along the coronal and sagittal axes, in patchwise fashion, with Residual U-Nets. All the estimated masks are then fed into a consensus module to produce a final volumetric segmentation of the nodule. The algorithm is rigorously evaluated on LIDC-IDRI dataset, which is the largest publicly available dataset. The proposed approach achieved the average dice score of 87.5%, which is significantly higher than the existing state-of-the-art techniques.

Outcome Differences of Remnant- Preserving versus Non-Preserving Methods in Arthroscopic Anterior Cruciate Ligament Reconstruction: A Meta-analysis with Subgroup analysis.

PURPOSE: To analyze differences in clinical outcomes of arthroscopic anterior cruciate ligament reconstruction between remnant-preserving and non-preserving methods. METHODS: International electronical databases PubMed, Embase, and the Cochrane central database from January 1966 to December 2017 were searched for randomized controlled trials (RCTs) and observational studies that compared differences of clinical outcomes of ACL reconstruction with and without remnant preservation. A meta-analysis of these studies was performed to compare clinical outcomes. Subgroup analyses were conducted to evaluate the role of methodological quality in primary meta-analysis estimates. RESULTS: Five RCTs and six observational studies were included in this meta-analysis and subgroup analysis. The remnant-preserving method in arthroscopic ACL reconstruction showed a statistically significant difference compared to the non-preserving method regarding arthrometric evaluation (side-to-side difference). Lachman test, Lysholm scores, and IKDC subjective scores showed statistically minor difference in meta-analysis, but showed no significant difference in subgroup analysis. Remained parameters including pivot shift test, IKDC grades, incidence of cyclops lesion showed no statistically differences in meta-analysis or subgroup analysis. CONCLUSIONS: This meta-analysis with subgroup analysis showed that arthroscopic remnant-preserving ACL reconstruction provided statistically significant but limited clinical relevance in terms of arthrometric evaluation. Results of Lachman test, Lysholm scores, and IKDC subjective scores demonstrated statistically minor differences.

Regulation of mRNA export through API5 and nuclear FGF2 interaction.

API5 (APoptosis Inhibitor 5) and nuclear FGF2 (Fibroblast Growth Factor 2) are upregulated in various human cancers and are correlated with poor prognosis. Although their physical interaction has been identified, the function related to the resulting complex is unknown. Here, we determined the crystal structure of the API5-FGF2 complex and identified critical residues driving the protein interaction. These findings provided a structural basis for the nuclear localization of the FGF2 isoform lacking a canonical nuclear localization signal and identified a cryptic nuclear localization sequence in FGF2. The interaction between API5 and FGF2 was important for mRNA nuclear export through both the TREX and eIF4E/LRPPRC mRNA export complexes, thus regulating the export of bulk mRNA and specific mRNAs containing eIF4E sensitivity elements, such as c-MYC and cyclin D1. These data show the newly identified molecular function of API5 and nuclear FGF2, and provide a clue to understanding the dynamic regulation of mRNA export.

Comparison of the Outcomes of Closed Reduction Nasal Bone Fractures With a Surgical Navigation System.

BACKGROUND: The most prevalent form of facial bone fractures is nasal fractures. The surgical procedures used for these fractures are relatively simple, but complete correction is not easy because the nasal bone is small and identifying the fracture site by palpation is difficult. This study aimed to investigate the efficacy of intraoperative surgical navigation systems in nasal bone fracture surgery through a prospective analysis. METHODS: Between February 2019 and July 2019, 25 navigation-assisted closed reductions of nasal fractures were performed. Preoperative computed tomography images were obtained at 1-mm intervals before surgery and the navigation was set by a simulation to have an error rate of less than 1. Then, the navigation system was used to identify the fracture site. Closed reduction was performed with Asch forceps and a Langenbeck elevator based on the previous markings made using the navigation system. RESULTS: The degree of reduction was evaluated by plain X-rays and computed tomography scans, which were performed 1 month after surgery. In the navigation group, the average distance between the fragment and normal bony alignment was decreased from 2.38 to 0.49 mm and the modified Motomura score was an average of 2.40 points. The decrease in the mean distance was significantly different (P = 0.038) compared with the conventional group. CONCLUSIONS: Surgical navigation systems could be a useful tool for localizing fracture sites and guiding closed reductions. In particular, the system could be recommended for nasal bone fracture reductions in the tip or pyriform regions, which are difficult to correct. LEVEL OF EVIDENCE: II.

Use of a biopsy punch for end-to-side anastomosis in free-tissue transfer.

Background: In end-to-side vessel-anastomoses, a side fenestration should be made by a slit incision or partial excision of the vessel wall. However, a slit incision might restrict blood flow across the anastomosis, and a partial vessel wall excision using micro-scissors may be time-consuming and result in irregular edges, which may weaken the vessel wall and cause flow disturbances. We used a biopsy punch for end-to-side anastomosis, and obtained satisfactory results.Methods: Between September 2015 and August 2017, 27 end-to-side anastomoses using punch biopsies were performed. Two vessel clamps were applied to the recipient vessel proximal and distal to the point planned for the side anastomosis. The flap side vessel size was measured, and an appropriately sized punch was selected. The clamped recipient vessel was supported by a wooden tongue depressor and the punch was applied to the vessel wall. An end-to-side anastomosis was performed in the usual manner, and immediate patency was tested with a refill test.Results: The vessel patency rate was 96 percent. The mean arteriotomy or venotomy time was 65 s and entire anastomosis procedure time was 1065 s. Both the vessel preparation and the micro-anastomosis procedure times were shorter than those of the conventional procedure group with statistical significance. (p < .05) There was only one case of venous failure by deep vein thrombosis - and it was resolved with thrombectomy and re-anastomosis.Conclusion: Use of a biopsy punch in end-to-side anastomosis can offer a uniform circular edge and reduce operating time.