Neoadjuvant camrelizumab plus apatinib for locally advanced microsatellite instability-high or mismatch repair-deficient colorectal cancer (NEOCAP): a single-arm, open-label, phase 2 study.

BACKGROUND: PD-1 blockade is highly efficacious for mismatch repair-deficient colorectal cancer in both metastatic and neoadjuvant settings. We aimed to explore the activity and safety of neoadjuvant therapy with PD-1 blockade plus an angiogenesis inhibitor and the feasibility of organ preservation in patients with locally advanced mismatch repair-deficient colorectal cancer. METHODS: We initiated a single-arm, open-label, phase 2 trial (NEOCAP) at Sun Yat-sen University Cancer Center and the Provincial Hospital of Traditional Chinese Medicine, Guangzhou, China. Patients aged 18-75 years with untreated mismatch repair-deficient or microsatellite instability-high or POLE/POLD1-mutated locally advanced colorectal cancer (cT3 or N+ for rectal cancer, and T3 with invasion ≥5mm or T4, with or without N+ for colon cancer) and an Eastern Cooperative Oncology Group performance score of 0-1 were enrolled and given 200 mg camrelizumab intravenously on day 1 and 250 mg apatinib orally from day 1-14, every 3 weeks for 3 months followed by surgery or 6 months if patients did not have surgery. Patients who had a clinical complete response did not undergo surgery and proceeded with a watch-and-wait approach. The primary endpoint was the proportion of patients with a pathological or clinical complete response. Eligible enrolled patients who received at least one cycle of neoadjuvant treatment and had at least one tumour response assessment following the baseline assessment were included in the activity analysis, and patients who received at least one dose of study drug were included in the safety analysis. The study is registered with ClinicalTrials.gov (NCT04715633) and is ongoing. FINDINGS: Between Sept 29, 2020, and Dec 15, 2022, 53 patients were enrolled; one patient was excluded from the activity analysis because they were found to be mismatch repair-proficient and microsatellite-stable. 23 (44%) patients were female and 29 (56%) were male. The median follow-up was 16·4 (IQR 10·5-23·5) months. 28 (54%; 95% CI 35-68) patients had a clinical complete response and 24 of these patients were managed with a watch-and-wait approach, including 20 patients with colon cancer and multiple primary colorectal cancer. 23 (44%) of 52 patients underwent surgery for the primary tumour, and 14 (61%; 95% CI 39-80) had a pathological complete response. 38 (73%; 95% CI 59-84) of 52 patients had a complete response. Grade 3-5 adverse events occurred in 20 (38%) of 53 patients; the most common were increased aminotransferase (six [11%]), bowel obstruction (four [8%]), and hypertension (four [8%]). Drug-related serious adverse events occurred in six (11%) of 53 patients. One patient died from treatment-related immune-related hepatitis. INTERPRETATION: Neoadjuvant camrelizumab plus apatinib show promising antitumour activity in patients with locally advanced mismatch repair-deficient or microsatellite instability-high colorectal cancer. Immune-related adverse events should be monitored with the utmost vigilance. Organ preservation seems promising not only in patients with rectal cancer, but also in those with colon cancer who have a clinical complete response. Longer follow-up is needed to assess the oncological outcomes of the watch-and-wait approach. FUNDING: The National Natural Science Foundation of China, Guangdong Basic and Applied Basic Research Foundation, and the Cancer Innovative Research Program of Sun Yat-sen University Cancer Center. TRANSLATION: For the Chinese translation of the abstract see Supplementary Materials section.

Non-enzymatic browning of wine induced by monomeric flavan-3-ols: A review.

Non-enzymatic browning occurs widely in both white and red wines, and it has a huge impact on the color evolution and aging potential. Previous studies have proved that phenolic compounds, especially those with catechol groups, are the most important substrates involved in browning reactions of wine. This review focus on the current knowledge of non-enzymatic browning in wine resulting from monomeric flavan-3-ols. First, some relevant aspects of monomeric flavan-3-ols are introduced, including their structures, origins, chemical reactivities, as well as potential impacts on the organoleptic properties of wine. Second, the mechanism for non-enzymatic browning induced by monomeric flavan-3-ols is discussed, with an emphasis on the formation of yellow xanthylium derivatives, followed by their spectral properties and effects on the color change of wine. Finally, attentions are also be given to the factors that influence non-enzymatic browning, such as metal ions, light exposure, additives in winemaking, etc.

Hairy gene homolog increases nasopharyngeal carcinoma cell stemness by upregulating Bmi-1.

B-cell-specific Moloney murine leukemia virus integration site 1 (Bmi-1) is overexpressed in various cancer types. We found that Bmi-1 mRNA levels were elevated in nasopharyngeal carcinoma (NPC) cell lines. In immunohistochemical analyses, high Bmi-1 levels were observed in not only 5 of 38 non-cancerous nasopharyngeal squamous epithelial biopsies, but also in 66 of 98 NPC specimens (67.3%). High Bmi-1 levels were detected more frequently in T3-T4, N2-N3 and stage III-IV NPC biopsies than in T1-T2, N0-N1 and stage I-II NPC samples, indicating that Bmi-1 is upregulated in advanced NPC. In 5-8F and SUNE1 NPC cells, stable depletion of Bmi-1 using lentiviral RNA interference greatly suppressed cell proliferation, induced G1-phase cell cycle arrest, reduced cell stemness and suppressed cell migration and invasion. Likewise, knocking down Bmi-1 inhibited NPC cell growth in nude mice. Both chromatin immunoprecipitation and Western blotting assays demonstrated that Hairy gene homolog (HRY) upregulated Bmi-1 by binding to its promoter, thereby increasing the stemness of NPC cells. Immunohistochemistry and quantitative real-time PCR analyses revealed that HRY expression correlated positively with Bmi-1 expression in a cohort of NPC biopsies. These findings suggested that HRY promotes NPC cell stemness by upregulating Bmi-1, and that silencing Bmi-1 can suppress NPC progression.

Sub-Regional Variation and Characteristics of Cabernet Sauvignon Wines in the Eastern Foothills of the Helan Mountain: A Perspective from Phenolics, Visual Properties and Mouthfeel.

As one of the most promising wine regions in China, the eastern foothills of the Helan Mountain (EFHM) in the Ningxia Hui Autonomous Region has attracted great attention recently. Geographically, EFHM is divided into six sub-regions, namely Shizuishan, Xixia, Helan, Qingtongxia, Yongning and Hongsipu. However, there have been few reports on the character and differences between wines in the six sub-regions. In this experiment, a total of 71 commercial Cabernet Sauvignon wines from six sub-regions were collected, and their phenolic compounds, visual properties and mouthfeel were investigated. The results showed that wines from the six sub-regions of EFHM showed distinctive phenolic profiles and could be distinguished through the OPLS-DA mode using 32 potential markers. In terms of color, Shizuishan wines showed higher a* values and lower b* values. The sensory evaluation showed that Hongsipu wines had higher astringency strength and lower tannin texture. The overall results implied that the phenolic compounds of wines in different sub-regions were affected by terroir conditions. To the best of our knowledge, this is the first time that a wide coverage of phenolic compounds has been analysed for wines from the sub-regions of EFHM, which could provide valuable information in deciphering the terroir of EFHM.

Artificial saliva precipitation index (ASPI): An efficient evaluation method of wine astringency.

Astringency is one of the most important organoleptic characteristics of red wines, and its intensity evaluation method has been the focus of research in recent years. An artificial saliva system was developed to establish an accurate and reliable evaluation method for the astringency intensity of dry red wines based on saliva precipitation index (SPI). To achieve this, five key protein families, which presented high reactivities and sensitivities in protein-tannin binding reactions, were selected from human whole saliva. The concentrations of the five proteins (proline-rich protein, α-amylase, lactoferrin, lysozyme, and albumin) and pH were optimized using response surface methodology based on the human salivary conditions to simulate the real salivary environment. The artificial saliva precipitation index method was applied to 60 commercial dry red wines and it exhibited a high correlation (CoefASPI = 0.94) with the sensory scores, indicating better performance than the traditional SPI method and other analytical approaches.

The compositional characteristics, influencing factors, effects on wine quality and relevant analytical methods of wine polysaccharides: A review.

Wine quality is closely related to various compounds including polysaccharides, a class of crucial macromolecules that affect its chemical and physical properties by influencing the colloidal state or interacting with other compounds via non-covalent bonds. Herein, the composition and structural characteristics of the major polysaccharides identified in wine and the factors influencing their contents are briefly described. An improved understanding of the molecular mechanisms of wine polysaccharides and their practical applications on wine stability and organoleptic qualities is thoroughly discussed. The effects of polysaccharides on wine quality are significantly correlated with their structure and composition as well as wine matrix composition. Thus, to better understand the chemical complexity of polysaccharides, relevant analytical methods are systematically summarized and highlighted, which may ultimately lead to the development of novel winery guidelines.

Genomic Analysis Uncovers the Prognostic and Immunogenetic Feature of Pyroptosis in Gastric Carcinoma: Indication for Immunotherapy.

Crosstalk between pyroptosis and tumor immune microenvironment (TIME) in cancer has yet to be elucidated. Herein, we aimed to explore the role of pyroptosis and its association with TIME in gastric cancer. Unsupervised clustering was performed to identify the pyroptosis-related clusters. Pyroptosis risk score was constructed using LASSO Cox regression. Clinicopathological and genetic data of pyroptosis clusters and pyroptosis risk scores were explored. Reproducibility of pyroptosis risk score in predicting response to immunotherapy and screening potential antitumor drugs was also investigated. Three pyroptosis clusters with distinct prognosis, immune cell fractions and signatures, were constructed. A low-pyroptosis risk score was characterized by increased activated T-cell subtype and M1 macrophage, decreased M2 macrophage, higher MSI status, and TMB. Meanwhile, low-score significantly correlated with PD-L1 expression, antigen presentation markers, and IFN-γ signature. The 5-year AUCs of PRS were 0.67, 0.62, 0.65, 0.67, and 0.67 in the TCGA, three external public and one real-world validation (SYSUCC) cohorts. Multivariable analyses further validated the prognostic performance of the pyroptosis risk scoring system, with HRs of 2.43, 1.83, 1.78, 2.35, and 2.67 (all p < 0.05) in the five cohorts. GSEA indicated significant enrichment of DNA damage repair pathways in the low-score group. Finally, the pyroptosis risk scoring system was demonstrated to be useful in predicting response to immunotherapy, and in screening potential antitumor drugs. Our study highlights the crucial role of interaction between pyroptosis and TIME in gastric cancer. The pyroptosis risk scoring system can be used independently to predict the survival of individuals and their response to immunotherapy.

Red wine coloration: A review of pigmented molecules, reactions, and applications.

Color is one of the most distinctive qualities of red wine. Despite new knowledge in the field of pigment identification, copigmentation, and oxidation being forthcoming, there is still a large gap between the fundamental research and practical winemaking outcomes. A state-of-art review from these two aspects is, therefore, necessary. This review first introduces updated knowledge about the primary pigments in wine, with emphasis on their physicochemical properties. Then, the mechanisms of copigmentation and oxidation are elucidated in detail, along with their relative contributions to wine color. Finally, the practical effects of copigmentation and micro-oxygenation (MOX) in winemaking are summarized and discussed. In general, wine coloration is ultimately determined by the anthocyanin flavylium cation, which is greatly influenced by wine pH. In young red wine, grape-derived anthocyanins and nonanthocyanin polyphenols (as copigments) are the foundation for wine coloration. During aging and storage, anthocyanin derivatives are formed via various chemical reactions, where moderate oxidation plays a vital role, whereas copigmentation constantly decreases. The essence of wine color evolution relates to the changes of physicochemical properties of primary pigments in wine, where the hydration equilibrium gradually diminishes. In practice, the effects of copigment addition and MOX during real vinification can be viewed as somewhat controversial, considering that many studies showed different effects on wine color and pigment concentration. Universal features can be summarized but some phenomena still remain unclear and deserve further exploration.

Iodine-catalyzed oxidative annulation: facile synthesis of pyrazolooxepinopyrazolones via methyl azaarene sp3 C-H functionalization.

An iodine-catalyzed methyl azaarene sp3 C-H functionalization has been developed for the synthesis of a seven-membered O-heterocyclic architecture containing three different heterocyclic aromatic hydrocarbons. This method can be applied to a wide range of substituted methyl azaarenes and diverse 2,4-dihydro-3H-pyrazol-3-ones, and brings about the efficient preparation of 2,9-dihydrooxepino[2,3-c:6,5-c']dipyrazol-3(7H)-ones in high yields with the merits of low catalyst loading, good functional group tolerance and metal-free conditions.

The formation mechanism of pinotin A in model wine: Experimental and theoretical investigation.

Pinotin A is a crucial pyranoanthocyanin in aged red wine. The formation mechanism of pinotin A was investigated through experimental and theoretical approaches in model wine solution. Using UHPLC-DAD-QTOF, three intermediates were identified in the reaction solution of malvidin-3-O-glucoside and caffeic acid. The pH and oxidative conditions affected the reaction rate by modulating the accumulation of intermediates. Upon preparative HPLC and oxidation experiment, the transformation of intermediate 3 to intermediate 2 was observed through free radical oxidation, p-benzoquinone oxidation and autooxidation. However, the production of pinotin A was only achieved through autooxidation. A three-steps reaction mechanism of reversible addition reaction and decarboxylation, and autooxidation was proposed. The validity of the proposed mechanism was verified through quantum chemistry calculation. The negative variation of Gibbs free energy of the entire reaction was obtained, where the autooxidation step played a crucial role for the spontaneity of the whole reaction.

Impact of three phenolic copigments on the stability and color evolution of five basic anthocyanins in model wine systems.

Phenolic copigments have important influence on red wine color. In this study, UV-visible spectrophotometer and UHPLC-Q-TOF-MS were combined to investigate the effects of three types of phenolic copigments (gallic acid, (-)-epicatechin, and quercetin-3-O-glucoside) on the stability and color properties of five common 3-O-monoglucosidic anthocyanins in model wine solutions. Results showed low concentrations (0.5 mM) of gallic acid and (-)-epicatechin protected anthocyanins from degradation, whereas high concentrations (8 mM) of them had the opposite effect. Quercetin-3-O-glucoside always improved the stability of anthocyanins despite its additive amount (0.1 mM or 0.4 mM). Even small quantity of (-)-epicatechin led to obvious yellow hue into the solution, and xanthylium derivatives generated from (-)-epicatechin were detected. Antagonistic effect among the three copigments was observed, probably as a result of competition of intermolecular copigmentation. Additionally, the stability of anthocyanins was significantly influenced by their structures: cyanidin-3-O-glucoside, peonidin-3-O-glucoside, and malvidin-3-O-glucoside were more stable than delphinidin-3-O-glucoside and petunidin-3-O-glucoside.

Genome-Wide Profiling Reveals HPV Integration Pattern and Activated Carcinogenic Pathways in Penile Squamous Cell Carcinoma.

Human papillomavirus (HPV) is a significant etiologic driver of penile squamous cell carcinoma (PSCC). The integration pattern of HPV and its carcinogenic mechanism in PSCC remain largely unclear. We retrospectively reviewed 108 PSCC cases who received surgery between 2008 and 2017. Using high-throughput viral integration detection, we identified 35 HPV-integrated PSCCs. Unlike cervical cancer, the HPV E2 oncogene was not prone to involvement in integration. Eleven of the 35 (31.4%) HPV-integrated PSCCs harbored intact HPV E2; these tumors had lower HPV E6 and E7 expression and higher expression of p53 and pRb proteins than those with disrupted E2 did (p < 0.001 and p = 0.024). Integration breakpoints are preferentially distributed in or near host genes, including previously reported hotspots (KLF5, etc.) and newly identified hotspots (CADM2, etc.), which are mainly involved in oncogenic signaling pathways (MAPK, JAK/STAT, etc.). Regarding the phosphorylation levels of JNK, p38 was higher in HPV-positive tumors with MAPK-associated integration than those in HPV-positive tumors with other integration and those in HPV-negative tumors. In vitro, KLF5 knockdown inhibited proliferation and invasion of PSCC cells, while silencing CADM2 promoted migration and invasion. In conclusion, this study enhances our understanding of HPV-induced carcinogenesis in PSCC, which may not only rely on the E6/E7 oncogenes, but mat also affect the expression of critical genes and thus activate oncogenic pathways.

Chitosan/Polyvinyl Alcohol/ Lauramidopropyl Betaine/2D-HOF Mixed Film with Abundant Hydrogen Bonds Acts as High Mechanical Strength Artificial Skin.

The mechanical properties of artificial skins are complicated to maintain under ensuring air permeability and antimicrobial. Thus, a series of hydrophilic antimicrobial polymer networks are prepared by crosslinking chitosan and polyvinyl alcohol with the lauramidopropyl betaine and hydrogen bond organic framework (CS/PVA/LPB/2D-HOF). The mechanical performance of the control groups and the complex are systematically evaluated to attain an artificial strength skin. The CS/PVA/LPB/2D-HOF complex exhibits strong mechanical abilities than other control groups. By analyzing the IR spectra and the morphology, the synergistic effect of hydrogen bonds between molecules and cracks significantly improves the mechanical properties of the complex. Its maximum tensile strength can reach 29 MPa, and its maximum load capacity can reach 3700 g. Notably, the composite membrane also performs an excellent antimicrobial activity. In vivo and in vitro experiments show that the hybrid membrane can promote tissue regeneration and wound healing (95％). These results may open up the opportunity for future composite material investigations in the artificial skin and tissue engineering field.

Comparative Analysis of Glycosidic Aroma Compound Profiling in Three Vitis vinifera Varieties by Using Ultra-High-Performance Liquid Chromatography Quadrupole-Time-of-Flight Mass Spectrometry.

Glycosidic aroma compounds are the important precursors of volatile aroma in grapes, and they can be added with odorous aglycones via enzyme- or acid-catalyzed hydrolysis during wine fermentation and storage. Developing an analytical method for intact glycosides can provide the possibility to study the accumulation of these aroma precursors in grape berries. For this purpose, a Tandem Mass Spectrometry (MS/MS). database based on ultra-high-performance liquid chromatography quadrupole-time-of-flight mass spectrometry was built, covering multiple aglycone classes. Subsequently, the profiles of glycosidic aroma compounds in Vitis vinifera L. cv. Muscat Blanc, Riesling, and Chardonnay berries during maturation were investigated. Pentosyl-hexosides were the most abundant glycosides in all three varieties. Both composition and concentration of glycosidic aroma compounds varied obviously among grape varieties. Except for monoterpenol pentosyl-hexosides, most glycosides were kept almost stable in their concentrations during berry maturation. This research provides an approach to understand the variation of glycosidic aroma components from the perspective of aglycones and glycosides.

Coexpression of CMTM6 and PD-L1 as a predictor of poor prognosis in macrotrabecular-massive hepatocellular carcinoma.

The "macrotrabecular-massive" (MTM) pattern of hepatocellular carcinoma (HCC) has been suggested to represent a distinct HCC subtype and is associated with specific molecular features. Since the immune microenvironment is heterogenous in HCC, it is important to evaluate the immune microenvironment of this novel variant. CMTM6, a key regulator of PD-L1, is an important immunocheckpoint inhibitor. This study aimed to evaluate the prognostic effect of CMTM6/PD-L1 coexpression and its relationship with inflammatory cells in HCC. We analyzed 619 HCC patients and tumors were classified into MTM and non-MTM HCC subtypes. The expression levels of CMTM6 and PD-L1 in tumor and inflammatory cells were evaluated by immunohistochemistry. The density of inflammatory cells in the cancer cell nest was calculated. Tumoral PD-L1 expression and inflammatory cell density were higher in the MTM type than in the non-MTM type. CMTM6-high expression was significantly associated with shorter OS and DFS than CMTM6-low expression in the whole HCC patient population and the MTM HCC patient population. Moreover, MTM HCC patients with CMTM6/PD-L1 coexpression experienced a higher risk of HCC progression and death. In addition, CMTM6/PD-L1 coexpression was shown to be related to a high density of inflammatory cells. Notably, a new immune classification, based on CMTM6/PD-L1 coexpression and inflammatory cells, successfully stratified OS and DFS in MTM HCC. CMTM6/PD-L1 coexpression has an adverse effect on the prognosis of HCC patients, especially MTM HCC patients. Our study provides evidence for the combination of immune status assessment with anti-CMTM6 and anti-PD-L1 therapy in MTM HCC patients.

Targeted metabolomics of anthocyanin derivatives during prolonged wine aging: Evolution, color contribution and aging prediction.

Anthocyanin derivatives and chromatic characteristics of 234 different-vintage red wine were investigated based on a targeted HPLC-MS/MS and CIELAB approach. The K-means cluster analysis showed that the evolution pattern varies amongst anthocyanin derivative classes. Their stabilities are: pinotins > flavanyl-pyranoanthocyanins, vitisin A > monomeric anthocyanin, direct anthocyanin-flavan-3-ols condensation products > vitisin B, anthocyanin ethyl-linked flavan-3-ols products. The proportion of most pyranoanthocyanins becomes more significant among all detected anthocyanin derivatives during wine aging, whereas flavanols-related anthocyanin derivatives (except for flavanyl-pyranoanthocyanins) decreased drastically. PLSR showed that aging tawny characteristics is related to pyranoanthocyanins except for vitisin B, especially pinotins, whereas monomeric anthocyanins and flavanol-related derivates (except for flavanyl-pyranoanthocyanins) contribute to red violet color. But aging color density is more associated with the content of vitisin A and flavanyl-pyranoanthocyanins. Two predictive models based on random forest and support vector machine modeling showed good performance in predicting the extent of wine aging.

Proposal for modified inguinofemoral lymphadenectomy derived from investigation of anatomic distribution of sentinel and metastatic nodes in vulvar cancer.

OBJECTIVE: We aimed to develop a less invasive inguinofemoral lymphadenectomy (IFL) approach for vulvar cancer based on the investigation of the anatomic distribution of sentinel and metastatic nodes. METHODS: Patients with vulvar cancer treated by surgery between 1995 and 2019 were retrospectively reviewed. A seven-field method was adopted to assign the anatomic locations for lymph nodes removed via IFL or sentinel node biopsy. Only patients with nodal metastasis or sentinel nodes were included. RESULTS: A total of 102 patients with eligible data were analyzed. Nodal metastasis was confirmed in 118 groins undergoing IFL; sentinel node detection succeeded in 46 groins. The medial-inguinal field had the highest rate of nodal metastasis involvement (59.3%, 70/118) and sentinel nodes present (73.9%, 34/46). The inferior-femoral field was involved only in one groin with quadruple-field metastases. The lateral-inguinal field was not involved in any groin. Neither the lateral-inguinal nor the inferior-femoral field presented sentinel nodes. CONCLUSION: The lateral-inguinal and inferior-femoral fields of the groins have a low risk of developing nodal metastasis. Therefore, a modified IFL preserving these fields can be established to reduce surgical morbidity without sacrificing its therapeutic effect.

AKR1C2 acts as a targetable oncogene in esophageal squamous cell carcinoma via activating PI3K/AKT signaling pathway.

The aldo-keto reductases family 1 member C2 (AKR1C2) has critical roles in the tumorigenesis and progression of malignant tumours. However, it was also discovered to have ambiguous functions in multiple cancers and till present, its clinical significance and molecular mechanism in oesophageal squamous cell carcinoma (ESCC) has been unclear. The aim of this study was to explore the role of AKR1C2 in the tumorigenesis of ESCC. Here, we showed that AKR1C2 expression was found to be up-regulated in ESCC tissues and was significantly associated with pathological stage, lymph node metastasis and worse outcomes. Functional assays demonstrated that an ectopic expression of AKR1C2 in ESCC cells resulted in increased proliferation, migration and cisplatin resistance, while knockdown led to inversing effects. Bioinformation analyses and mechanistic studies demonstrated that AKR1C2 activated the PI3K/AKT signalling pathway, furthermore, the inhibitor of PI3K or the selective inhibitor of AKR1C2 enzyme activity could reverse the aggressiveness and showed synergistic antitumour effect when combined with cisplatin, both in vitro and in vivo. In conclusion, Our findings revealed that AKR1C2 could function as an oncogene by activating the PI3K/AKT pathway, as a novel prognostic biomarker and/or as a potential therapeutic target to ESCC.

Sectioning protocol determines accuracy of intraoperative pathological examination of sentinel lymph node in cervical cancer: A systematic review and meta-analysis.

OBJECTIVES: To determine the diagnostic performance and optimal protocol of frozen section examination (FSE) in SLNB for cervical cancer. METHODS: PubMed, EMBASE, Web of Science, Cochrane Library, Wanfang Data and China National Knowledge Infrastructure were searched from inception to July 30, 2019, for studies concerning SLNB with FSE in cervical cancer. Sensitivity of FSE in detecting SLN metastasis was the primary diagnostic indicator for evaluation. RESULTS: The pooled sensitivity of FSE among 31 eligible studies (1887 patients) was 0.77 (95% CI 0.66-0.85) with high heterogeneity (I2 = 69.73%). Two representative sectioning protocols for FSE were identified from 26 studies, described as equatorial (E-protocol, SLN was bisected) and latitudinal (L-protocol, SLN was cut at intervals). Meta-regression showed that FSE protocol was the only source of heterogeneity (p < 0.001). The pooled sensitivity was 0.86 (95% CI 0.79-0.91, I2 = 0%) and 0.59 (0.46-0.72, I2 = 58.47%) for FSE using L- (13 studies, 650 patients) and E- (13 studies, 1047 patients) protocol, respectively. Among the available data, marcometastases (>2 mm) were missed in 4 and 20 patients; small-volume metastases (≤2 mm) were detected in 13 and 2 patients, respectively, under L- and E-protocol. The pooled sensitivity of FSE using L-protocol would reach 0.97 (95% CI 0.89-0.99) if only marcometastases were considered. These findings were robust to sensitivity analyses. CONCLUSION: The sectioning protocol determines the accuracy of FSE in SLNB. With L-protocol, FSE can provide precise intraoperative pathology for SLNB, which enables immediate decision-making for individualized managements.

HPLC-MS/MS-based targeted metabolomic method for profiling of malvidin derivatives in dry red wines.

Anthocyanin derivatives are critical components that impart color to aging red wine. In this study, we developed a targeted metabolomic method for the simultaneously profiling of the primary thirty-seven malvidin-derived anthocyanin derivatives in red wine, including various pyranoanthocyanins and flavanols-related condensation products. First, high-performance liquid chromatography (HPLC) tandem ion trap and triple-quadrupole (QqQ) mass spectrometry were used to construct the mass spectral and chromatographic database of the anthocyanin derivatives that were formed in a model wine solution. Next, the targeted profiling analysis of these compounds was achieved on a QqQ mass spectrometer in the multiple reaction monitoring mode (MRM). The method displayed excellent linearity (R2 0.9391-0.9998), sensitivity (0.221-0.604 µg/L of limit of detection (LOD) and 0.274-1.157 µg/L of limit of quantification (LOQ) equivalent to malvidin-3-O-glucoside (Mv-glc)), and repeatability (less than 10% and 15% for intra-day and inter-day relative standard deviation (RSD) respectively). Partial least squares discriminant analysis (PLS-DA) based on this method showed great discrimination over different vintage wines, thereby promising to be an effective tool in wine anthocyanin and aging related study.