Heterologous prime-boost vaccination drives early maturation of HIV broadly neutralizing antibody precursors in humanized mice.

A protective HIV vaccine will likely need to induce broadly neutralizing antibodies (bnAbs). Vaccination with the germline-targeting immunogen eOD-GT8 60mer adjuvanted with AS01B was found to induce VRC01-class bnAb precursors in 97% of vaccine recipients in the IAVI G001 phase 1 clinical trial; however, heterologous boost immunizations with antigens more similar to the native glycoprotein will be required to induce bnAbs. Therefore, we designed core-g28v2 60mer, a nanoparticle immunogen to be used as a first boost after eOD-GT8 60mer priming. We found, using a humanized mouse model approximating human conditions of VRC01-class precursor B cell diversity, affinity, and frequency, that both protein- and mRNA-based heterologous prime-boost regimens induced VRC01-class antibodies that gained key mutations and bound to near-native HIV envelope trimers lacking the N276 glycan. We further showed that VRC01-class antibodies induced by mRNA-based regimens could neutralize pseudoviruses lacking the N276 glycan. These results demonstrated that heterologous boosting can drive maturation toward VRC01-class bnAb development and supported the initiation of the IAVI G002 phase 1 trial testing mRNA-encoded nanoparticle prime-boost regimens.

Disulfiram Improves Fat Graft Retention by Modulating Macrophage Polarization With Inhibition of NLRP3 Inflammasome-Mediated Pyroptosis.

BACKGROUND: Macrophage-mediated inflammatory response in the early post-grafting period restricts fat graft retention. Pyroptosis is a novel type of programmed cell death that extensively participates in inflammatory pathologies. OBJECTIVES: This study sought to determine whether macrophage pyroptosis was activated during the inflammatory phase after fat grafting and to investigate the efficacy of a pyroptosis inhibitor, disulfiram (DSF), in fat graft retention. METHODS: We established a C57BL/6 mice fat grafting model and then analyzed macrophage pyroptosis. DSF (50 mg/kg, every other day) was intraperitoneally injected starting 1 hour before fat grafting and continued for 14 days. An in vitro co-culture system was established in which mouse RAW264.7 macrophages were co-cultured with apoptotic adipocytes to further validate the findings of the in vivo studies and to explore the underlying mechanisms. RESULTS: Here we reported that macrophage pyroptosis was activated in both fat grafts and in vitro co-culture models. DSF was found to be a potent pyroptosis inhibitor, promoting M2 macrophage polarization. In addition, DSF was demonstrated to enhance vascularization and graft retention. CONCLUSIONS: Our results suggested that pyroptosis plays a crucial role in the inflammatory cascade within fat grafts. DSF, being a clinically available drug, could be translated into a clinically effective drug for improving fat graft survival by inhibiting macrophage pyroptosis, therefore inducing M2 macrophage polarization and promoting neovascularization.

TrieDedup: a fast trie-based deduplication algorithm to handle ambiguous bases in high-throughput sequencing.

BACKGROUND: High-throughput sequencing is a powerful tool that is extensively applied in biological studies. However, sequencers may produce low-quality bases, leading to ambiguous bases, 'N's. PCR duplicates introduced in library preparation are conventionally removed in genomics studies, and several deduplication tools have been developed for this purpose. Two identical reads may appear different due to ambiguous bases and the existing tools cannot address 'N's correctly or efficiently. RESULTS: Here we proposed and implemented TrieDedup, which uses the trie (prefix tree) data structure to compare and store sequences. TrieDedup can handle ambiguous base 'N's, and efficiently deduplicate at the level of raw sequences. We also reduced its memory usage by approximately 20% by implementing restrictedDict in Python. We benchmarked the performance of the algorithm and showed that TrieDedup can deduplicate reads up to 270-fold faster than pairwise comparison at a cost of 32-fold higher memory usage. CONCLUSIONS: The TrieDedup algorithm may facilitate PCR deduplication, barcode or UMI assignment, and repertoire diversity analysis of large-scale high-throughput sequencing datasets with its ultra-fast algorithm that can account for ambiguous bases due to sequencing errors.

CRISPR/Cas9-Mediated Knockout of the HuR Gene in U251 Cell Inhibits Japanese Encephalitis Virus Replication.

Human antigen R (HuR) is an RNA-binding protein that regulates the post-transcriptional reaction of its target mRNAs. HuR is a critical factor in cancer development and has been identified as a potential target in many cancer models. It participates in the viral life cycle by binding to viral RNAs. In prior work, we used CRISPR/Cas9 screening to identify HuR as a prospective host factor facilitating Japanese encephalitis virus (JEV) infection. The HuR gene was successfully knocked out in U251 cell lines using the CRISPR/Cas9 gene-editing system, with no significant difference in cell growth between U251-WT and U251-HuR-KO2 cells. Here, we experimentally demonstrate for the first time that the knockout of the HuR gene inhibits the replication ability of JEV in U251 cell lines. These results play an essential role in regulating the replication level of JEV and providing new insights into virus-host interactions and potential antiviral strategies. It also offers a platform for investigating the function of HuR in the life cycle of flaviviruses.

Regulation of Multiple Resonance Delayed Fluorescence via Through-Space Charge Transfer Excited State towards High-Efficiency and Stable Narrowband Electroluminescence.

B- and N-embedded multiple resonance (MR) type thermally activated delayed fluorescence (TADF) emitters usually suffer from slow reverse intersystem crossing (RISC) process and aggregation-caused emission quenching. Here, we report the design of a sandwich structure by placing the B-N MR core between two electron-donating moieties, inducing through-space charge transfer (TSCT) states. The proper adjusting of the energy levels brings about a 10-fold higher RISC rate in comparison with the parent B-N molecule. In the meantime, a high photoluminescence quantum yield of 91 % and a good color purity were maintained. Organic light-emitting diodes based on the new MR emitter achieved a maximum external quantum efficiency of 31.7 % and small roll-offs at high brightness. High device efficiencies were also obtained for a wide range of doping concentrations of up to 20 wt % thanks to the steric shielding of the B-N core. A good operational stability with LT95 of 85.2 h has also been revealed. The dual steric and electronic effects resulting from the introduction of a TSCT state offer an effective molecular design to address the critical challenges of MR-TADF emitters.

Precisely regulating the double-boron-based multi-resonance framework towards pure-red emitters: high-performance OLEDs with CIE coordinates fully satisfying the BT. 2020 standard.

Here, we propose a new simple and effective strategy for designing pure-red multi-resonance (MR) emitters through precisely regulating the double-boron-based MR framework. The two designed emitters exhibit ultrapure red emission together with superb photophysical properties, and further enable high-performance, high color-purity red OLEDs.

Facile synthesis of defect-rich RuCu nanoflowers for efficient hydrogen evolution reaction in alkaline media.

Developing high-performance electrocatalysts toward hydrogen evolution reaction (HER) in alkaline media is highly desirable for industrial applications in the field of water splitting but is still challenging. Herein, we successfully synthesized RuCu nanoflowers (NFs) with tunable atomic ratios using a facile wet chemistry method. The Ru3Cu NFs need only 55 mV to achieve a current density of 10 mA cm-2, which shows ideal durability with only 4 mV decay after 2000 cycles, largely outperforming the catalytic properties of commercial Pt/C. The Ru3Cu NFs comprise many nanosheets that can provide more active sites for HER. In addition, the introduction of Cu can modulate the electronic structure of Ru, facilitate water dissociation, and optimize H adsorption/desorption ability. Thus, the flower-like structure together with the proper incorporation of Cu boosts HER performance.

Metal-Perturbed Multiresonance TADF Emitter Enables High-Efficiency and Ultralow Efficiency Roll-Off Nonsensitized OLEDs with Pure Green Gamut.

Multiresonance (MR)-induced thermally activated delayed fluorescence (TADF) emitters based on B- and N-embedded polycyclic aromatics are desirable for ultrahigh-definition organic light-emitting diodes (OLEDs) due to their high photoluminescence quantum yield (PLQY) and narrow bandwidth. But the reverse intersystem crossing (RISC) rates of MR-TADF emitters are usually small, resulting in severe device efficiency roll-off at high brightness. To solve this issue, a sensitizer for the MR-TADF emitter has been required. Herein, a new MR-TADF emitter is developed through coordination of Au with B/N-embedded polycyclic ligand. Benefitting from the Au perturbation, the RISC rate is dramatically accelerated to 2.3 × 107 s-1 , leading to delayed fluorescence lifetime as short as 4.3 µs. Meanwhile, the PLQY of 95% and full width at half maximum of 39 nm (0.18 eV) are essentially unchanged after metal coordination. Therefore, a high PLQY, short delayed fluorescence lifetime, and high color purity are concurrently realized in a single TADF emitter. Accordingly, vacuum-deposited OLEDs exhibit high-performance electroluminescence with a maximum external quantum efficiency (EQE) of 35.8% without sensitization. The EQE is maintained as high as 32.3% at 10 000 cd m-2 . Furthermore, solution-processed OLED based on the emitter also achieves excellent performance with a maximum EQE of 25.7% and a small efficiency roll-off.

Application of 3-Dimensional Technology for Evaluating Muscular Type and Muscle-Fat Pad Mixed-Type Nasolabial Folds With Botulinum Toxin-A Treatment.

BACKGROUND: Botulinum toxin-A (BTX-A) is used in the treatment of nasolabial folds (NLFs). However, lighting and clinician subjectivity play a major role in evaluating the efficacy of this treatment. OBJECTIVES: By applying 3-dimensional (3D) technology, this study aimed to quantitatively evaluate the effects of BTX-A injection on muscular (M) and muscle-fat pad mixed-type (MF) NLFs. METHODS: BTX-A was injected into bilateral marked points on the NLFs, where the levator labii alaeque nasi, zygomaticus minor, and zygomaticus major pull the skin to form the NLF (2 U at each injection site). Pretreatment and posttreatment 3D facial images were captured with static and laughing expressions. The curvature, width, depth, and lateral fat volume of the NLFs were measured to compare the therapeutic efficacy for type M and MF NLFs. RESULTS: Thirty-nine patients with type M and 37 with type MF NLFs completed the follow-up data. In these patients, the curvature, width, and depth of the NLF showed a significant reduction at 1 month and gradually recovered at 3 and 6 months after treatment, with more significant improvement when laughing than when static. Variations compared to the pretreatment values of type MF were greater than those of type M at each time point. The lateral fat volume of the type MF NLF was significantly reduced (P < .05). CONCLUSIONS: 3D technology can quantitatively evaluate the effects BTX-A injection for treating type M and type MF NLFs. BTX-A is more effective on type MF than on type M NLFs.

The Retention-Rate Improvement of Stromal Vascular Fraction Gel in Prefrontal Filling With Botulinum Toxin-A Injection: A Retrospective Analysis.

BACKGROUND: As a derivative of adipose tissues, stromal vascular fraction gel has been widely utilized in facial soft tissue filling, but it still does not achieve the expected effect in forehead filling. The reason may be related to the corrugator muscles movements. OBJECTIVES: The authors aimed to evaluate the effect of botulinum toxin-A (BTX-A) on the retention rate of stromal vascular fraction gel by limiting the corrugator muscles movements and to provide a theoretical basis that short-term inhibition of movement in the affected area could improve the effects of the fat graft. METHODS: From January 2019 to June 2021, patients with stromal vascular fraction gel facial filling (including frontal and temporal parts) were selected. According to whether or not BTX-A treatment was received, patients were divided into injected and the noninjected groups. A questionnaire and the Global Aesthetic Improvement Scale (GAIS) were administered to evaluate 2-dimensional photos. The retention rate and curvature were calculated with 3-dimensional images utilizing Artec Studio 13 Professional and MATLAB software. RESULTS: The graft retention, forehead curvature, and GAIS scores were all higher in the injected group than the noninjected group (P < .01). On the questionnaire, the injected group also showed more satisfaction with the treatment effect and were more willing to recommend the treatment to their friends. CONCLUSIONS: BTX-A injection can improve the retention rate of prefrontal stromal vascular fraction gel filling, with higher patient satisfaction and better postoperative effects.

Humanized V(D)J-rearranging and TdT-expressing mouse vaccine models with physiological HIV-1 broadly neutralizing antibody precursors.

Antibody heavy chain (HC) and light chain (LC) variable region exons are assembled by V(D)J recombination. V(D)J junctional regions encode complementarity-determining-region 3 (CDR3), an antigen-contact region immensely diversified through nontemplated nucleotide additions ("N-regions") by terminal deoxynucleotidyl transferase (TdT). HIV-1 vaccine strategies seek to elicit human HIV-1 broadly neutralizing antibodies (bnAbs), such as the potent CD4-binding site VRC01-class bnAbs. Mice with primary B cells that express receptors (BCRs) representing bnAb precursors are used as vaccination models. VRC01-class bnAbs uniformly use human HC VH1-2 and commonly use human LCs Vκ3-20 or Vκ1-33 associated with an exceptionally short 5-amino-acid (5-aa) CDR3. Prior VRC01-class models had nonphysiological precursor levels and/or limited precursor diversity. Here, we describe VRC01-class rearranging mice that generate more physiological primary VRC01-class BCR repertoires via rearrangement of VH1-2, as well as Vκ1-33 and/or Vκ3-20 in association with diverse CDR3s. Human-like TdT expression in mouse precursor B cells increased LC CDR3 length and diversity and also promoted the generation of shorter LC CDR3s via N-region suppression of dominant microhomology-mediated Vκ-to-Jκ joins. Priming immunization with eOD-GT8 60mer, which strongly engages VRC01 precursors, induced robust VRC01-class germinal center B cell responses. Vκ3-20-based responses were enhanced by N-region addition, which generates Vκ3-20-to-Jκ junctional sequence combinations that encode VRC01-class 5-aa CDR3s with a critical E residue. VRC01-class-rearranging models should facilitate further evaluation of VRC01-class prime and boost immunogens. These new VRC01-class mouse models establish a prototype for the generation of vaccine-testing mouse models for other HIV-1 bnAb lineages that employ different HC or LC Vs.

Adipose-derived stem cells regulate CD4+ T-cell-mediated macrophage polarization and fibrosis in fat grafting in a mouse model.

Autologous fat grafting is becoming increasingly common worldly. However, the long-term retention of fat grafting is still unpredictable due to the inevitable fibrosis arising during tissue repair. Fibrosis may be regulated by T-cell immune responses that are influenced by adipose-derived stem cells (ASCs). Therefore, we hypothesized that overly abundant ASCs might promote fibrosis by promoting T-cell immune responses to adipose tissue. We performed 0.3 ml fat grafts with 104/ml, 106/ml and 108/ml ASCs and control group in C57 BL/6 mice in vivo. We observed retention, fibrosis, T-cell immunity, and macrophage infiltration over 12 weeks. Besides, CD4+ T-helper 1 (Th1) cells and T-helper 2 (Th2) cells were co-cultured with ASCs or ASCs conditioned media (ASCs-CM) in vitro. We detected the ratio of Th2%/Th1%. Results showed that the retention rate was higher in 104 group, while even lower in 108 group with significantly increased inflammation and fibrosis than control group at week 12 in vivo. There was no significance between control group and 106 group. Also, 108 group increased the infiltration of M2 macrophages, CD4+ T-cells and Th2/Th1 ratio. In vitro, the ratio of Th2%/Th1% induced by ASCs-transwell group was higher than ASCs-CM group and showed concentration-dependent. Accordingly, high concentrations of ASCs in adipose tissue can promote Th1-Th2 shifting, and excessive Th2 cells might promote the persistence of M2 macrophages and increase the level of fibrosis which lead to a decrease in the long-term retention of fat grafts. Also, we found ASCs promoted Th1-Th2 shifting in vitro.

The clinical value of neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, and D-dimer-to-fibrinogen ratio for predicting pneumonia and poor outcomes in patients with acute intracerebral hemorrhage.

Background: This study aimed to investigate the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and D-dimer-to-fibrinogen ratio (DFR) as predictors of pneumonia and poor outcomes in patients with acute intracerebral hemorrhage (ICH). Methods: We retrospectively examined patients with acute ICH treated in our institution from May 2018 to July 2020. Patient characteristics, laboratory testing data, radiologic imaging data, and 90-day outcomes were recorded and analyzed. Results: Among the 329 patients included for analysis, 183 (55.6%) developed pneumonia. Systolic blood pressure, initial hematoma volume, D-dimer concentration, NLR, PLR, DFR, and white blood cell, platelet, neutrophil, and lymphocyte counts at admission were significantly higher in patients who developed pneumonia than in those who did not; however, the Glasgow coma scale (GCS) score at admission was significantly lower in pneumonia patients compared with non-pneumonia patients (all P <0.05). Multivariate logistic regression showed that the NLR and PLR were independent predictors of pneumonia, and the NLR and DFR were independent predictors of poor 90-day outcomes (modified Rankin scale score 4-6). Conclusion: The NLR and PLR were independent predictors of pneumonia and the NLR and DFR were independent predictors of poor 90-day outcomes. The NLR, PLR, and DFR can provide prognostic information about acute ICH patients.

An antibody from single human VH-rearranging mouse neutralizes all SARS-CoV-2 variants through BA.5 by inhibiting membrane fusion.

SARS-CoV-2 Omicron subvariants have generated a worldwide health crisis due to resistance to most approved SARS-CoV-2 neutralizing antibodies and evasion of vaccination-induced antibodies. To manage Omicron subvariants and prepare for new ones, additional means of isolating broad and potent humanized SARS-CoV-2 neutralizing antibodies are desirable. Here, we describe a mouse model in which the primary B cell receptor (BCR) repertoire is generated solely through V(D)J recombination of a human VH1-2 heavy chain (HC) and, substantially, a human Vκ1-33 light chain (LC). Thus, primary humanized BCR repertoire diversity in these mice derives from immensely diverse HC and LC antigen-contact CDR3 sequences generated by nontemplated junctional modifications during V(D)J recombination. Immunizing this mouse model with SARS-CoV-2 (Wuhan-Hu-1) spike protein immunogens elicited several VH1-2/Vκ1-33-based neutralizing antibodies that bound RBD in a different mode from each other and from those of many prior patient-derived VH1-2-based neutralizing antibodies. Of these, SP1-77 potently and broadly neutralized all SARS-CoV-2 variants through BA.5. Cryo-EM studies revealed that SP1-77 bound RBD away from the receptor-binding motif via a CDR3-dominated recognition mode. Lattice light-sheet microscopy-based studies showed that SP1-77 did not block ACE2-mediated viral attachment or endocytosis but rather blocked viral-host membrane fusion. The broad and potent SP1-77 neutralization activity and nontraditional mechanism of action suggest that it might have therapeutic potential. Likewise, the SP1-77 binding epitope may inform vaccine strategies. Last, the type of humanized mouse models that we have described may contribute to identifying therapeutic antibodies against future SARS-CoV-2 variants and other pathogens.

Adipose-derived stem cells attenuate acne-related inflammation via suppression of NLRP3 inflammasome.

BACKGROUND: Acne is a chronic facial disease caused by Propionibacterium acnes, which proliferates within sebum-blocked skin follicles and increases inflammatory cytokine production. Several therapeutic drugs and products have been proposed to treat acne, yet no single treatment that ensures long-term treatment efficacy for all patients is available. Here, we explored the use of facial autologous fat transplant of adipose-derived stem cells (ADSCs) to dramatically reduce acne lesions. METHODS: THP-1 cells were treated with active P. acnes for 24 h at different multiplicities of infection, and alterations in inflammatory factors were detected. To study the effect of THP-1 on inflammasome-related proteins, we first co-cultured ADSCs with THP-1 cells treated with P. acnes and evaluated the levels of these proteins in the supernatant. Further, an acne mouse model injected with ADSCs was used to assess inflammatory changes. RESULTS: Propionibacterium acnes-mediated stimulation of THP-1 cells had a direct correlation with the expression of active caspase-1 and interleukin (IL)-1ß in an infection-dependent manner. ADSCs significantly reduced the production of IL-1ß induced by P. acnes stimulation through the reactive oxygen species (ROS)/Nod-like receptor family pyrin domain-containing 3 (NLRP3)/caspase-1 pathway. The results showed that ADSCs inhibit the skin inflammation induced by P. acnes by blocking the NLRP3 inflammasome via reducing the secretion of IL-1ß in vivo. CONCLUSIONS: Our findings suggest that ADSCs can alter IL-1ß secretion by restricting the production of mitochondria ROS, thereby inhibiting the NLRP3/caspase-1 pathway in P. acnes-induced inflammatory responses. This study indicates that anti-acne therapy can potentially be developed by targeting the NLRP3 inflammasome.

Effect of Positional Changes on Skin Landmarks in Midface Filling.

BACKGROUND: The design lines for midfacial filling shift upward with a patient's position changes from upright to supine during operation. This will cause the actual filled part to deviate from the target area. OBJECTIVES: This authors aimed to evaluate the effect of positional changes on midfacial landmarks and find the optimal body position for midface filling. METHODS: The process involved the grading and evaluation stages. The midfacial laxity of each sample in the evaluation stage was graded into minimal, moderate, and severe by the system established in the grading stage. Measured through the 3-dimensional images in each grade, the vertical distances from landmarks C, D, and E (representing the region of the tear trough, infraorbital area, and nasolabial fat pad, respectively) to the horizontal line of the inner canthus and depth of nasolabial fold at an angle of 90° were separately compared with those from the other angles (60°, 45°, 30°, and 0°) of the operating table. RESULTS: In the minimal midfacial laxity group, all 3 landmarks significantly moved upward when the angle decreased to 30°. However, landmark E of the moderate and severe and landmark D of the severe midfacial laxity groups both significantly moved upward when the angle decreased to 45°. The depth of the nasolabial fold at a 45° angle was significantly less than that at a 90° angle in the moderate and severe groups. CONCLUSIONS: In midface filling, a patient's body position should be optimally selected according to the midfacial laxity and filling area.

Application of Three-Dimensional Technology in Evaluating the Lower Face Lifting by Regional Platysma Injection with Botulinum Toxin-A.

BACKGROUND: Botulinum toxin-A (BTX-A) injection of regional platysma has been utilized in the lower-part elevation and mandibular contour sculpture. However, the relative research, especially in quantitative assessment appears very spare. Our aim is to investigate the efficacy of three-dimensional (3D) technology as a method for regional platysma injection with BTX-A. MATERIALS AND METHODS: From October 2019 to September 2020, patients with mild or moderate degrees of facial sagging on the lower face were recruited to regional platysma BTX-A injection, and 3D scanning and measurement technology was used to evaluate the difference of curved distances and angels. Patients' improvement was assessed by the global aesthetic improvement scale (GAIS). RESULTS: A total of 57 patients underwent regional platysma BTX-A injection and 32 patients were followed up successfully. Compared with Pre-operative, postoperative facial reference curves distance and cervico-mental angles had statistical differences (p < 0.05). GAIS suggested that the 3D imaging measurement technology could improve satisfaction. CONCLUSION: 3D technology can evaluate the improvement of the lower face with BTX-A. It provides effective measurement methods and raises satisfaction. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Long-term oncologic safety of immediate reconstructive surgery in patients with invasive breast cancer: a retrospective matched-cohort study.

OBJECTIVE: Immediate reconstruction (IR) is a safe and effective surgical treatment for patients with breast cancer. We aimed to assess the prognosis, aesthetic outcomes, and patient satisfaction of IR compared with breast conservation surgery (BCS) and total mastectomy (TM). METHODS: This retrospective matched-cohort study was conducted between May 2005 and December 2014. We established two cohorts according to the tumor (T) size of breast cancer. In the T≤3cm group, cases (IR) and controls (BCS or TM) were matched for age, pathological tumor size, and pathologic nodal status in a 1:1:1 ratio. In the T>3cm group, cases (IR) and controls (TM) were matched with the same factors and ratio. The primary outcome was the 5-year disease-free survival (DFS). The secondary outcome was patient satisfaction and quality of life. RESULTS: A total of 12,678 breast cancer patients were assessed for eligibility, of which 587 were included (T≤3 cm group: 155 IR vs 155 BCS vs 155 TM; T>3cm group: 61 IR vs 61 TM). In the T≤3 cm cohort, patients who underwent IR had no difference compared with those who underwent BCS or TM regarding the 5-year DFS (P=0.539); however, an improved aesthetic satisfaction, psychosocial, and sexual well-being were achieved in the IR group (P<0.001). In the T>3 cm cohort, the IR group had a worse median 5-year DFS (P=0.044), especially for Her2+ or triple-negative breast carcinoma (TNBC) subtypes compared with the TM group. CONCLUSIONS: IR improves aesthetic satisfaction, psychosocial, and sexual well-being for breast cancer patients with T≤3 cm. For patients with T > 3 cm invasive breast cancer, TM is superior to IR as it predicts a better 5-year DFS.