Atomic layer deposition (ALD)-constructed TaS2nanoflakes for cancer-related nucleolin detection.

Sensitive detection of nucleolin (NCL) is of great significance for the early diagnosis of cancer. In this work, as a new type of two-dimensional (2D) transition metal dichalcogenides (TMDCs), TaS2nanoflakes (NFs) were precisely constructed by atomic layer deposition (ALD) on carbon fiber paper (CFP) with high specific surface area.In situobservation showed that the nucleation and growth of TaS2nanoflakes were precisely controlled by the number of ALD cycles, thereby regulating their electrochemical properties. The electrochemical performance of TaS2NFs was observed in depth, and compared with that of traditional 2D TMDCs. Due to the high surface area and conductivity, anodic/cathodic current of ∼1570µA of TaS2NFs/CFP can be obtained. Subsequently, an electrochemical biosensor based on ALD-constructed TaS2NFs/CFP for cancer-related NCL detection was fabricated. Due to the excellent electrochemical performance of TaS2NFs/CFP, ultrasensitive detection of NCL in the linear range of 0.1 pM-10 nM with a detection limit of 0.034 pM was achieved.

Somatic mutations in collagens are associated with a distinct tumor environment and overall survival in gastric cancer.

BACKGROUND: Gastric cancer is a heterogeneous disease with poorly understood genetic and microenvironmental factors. Mutations in collagen genes are associated with genetic diseases that compromise tissue integrity, but their role in tumor progression has not been extensively reported. Aberrant collagen expression has been long associated with malignant tumor growth, invasion, chemoresistance, and patient outcomes. We hypothesized that somatic mutations in collagens could functionally alter the tumor extracellular matrix. METHODS: We used publicly available datasets including The Tumor Cancer Genome Atlas (TCGA) to interrogate somatic mutations in collagens in stomach adenocarcinomas. To demonstrate that collagens were significantly mutated above background mutation rates, we used a moderated Kolmogorov-Smirnov test along with combination analysis with a bootstrap approach to define the background accounting for mutation rates. Association between mutations and clinicopathological features was evaluated by Fisher or chi-squared tests. Association with overall survival was assessed by Kaplan-Meier and the Cox-Proportional Hazards Model. Gene Set Enrichment Analysis was used to interrogate pathways. Immunohistochemistry and in situ hybridization tested expression of COL7A1 in stomach tumors. RESULTS: In stomach adenocarcinomas, we identified individual collagen genes and sets of collagen genes harboring somatic mutations at a high frequency compared to background in both microsatellite stable, and microsatellite instable tumors in TCGA. Many of the missense mutations resemble the same types of loss of function mutations in collagenopathies that disrupt tissue formation and destabilize cells providing guidance to interpret the somatic mutations. We identified combinations of somatic mutations in collagens associated with overall survival, with a distinctive tumor microenvironment marked by lower matrisome expression and immune cell signatures. Truncation mutations were strongly associated with improved outcomes suggesting that loss of expression of secreted collagens impact tumor progression and treatment response. Germline collagenopathy variants guided interpretation of impactful somatic mutations on tumors. CONCLUSIONS: These observations highlight that many collagens, expressed in non-physiologically relevant conditions in tumors, harbor impactful somatic mutations in tumors, suggesting new approaches for classification and therapy development in stomach cancer. In sum, these findings demonstrate how classification of tumors by collagen mutations identified strong links between specific genotypes and the tumor environment.

A Vision-Based Social Distancing and Critical Density Detection System for COVID-19.

Social distancing (SD) is an effective measure to prevent the spread of the infectious Coronavirus Disease 2019 (COVID-19). However, a lack of spatial awareness may cause unintentional violations of this new measure. Against this backdrop, we propose an active surveillance system to slow the spread of COVID-19 by warning individuals in a region-of-interest. Our contribution is twofold. First, we introduce a vision-based real-time system that can detect SD violations and send non-intrusive audio-visual cues using state-of-the-art deep-learning models. Second, we define a novel critical social density value and show that the chance of SD violation occurrence can be held near zero if the pedestrian density is kept under this value. The proposed system is also ethically fair: it does not record data nor target individuals, and no human supervisor is present during the operation. The proposed system was evaluated across real-world datasets.

Knockdown of macrophage migration inhibitory factor (MIF), a novel target to protect neurons from parthanatos induced by simulated post-spinal cord injury oxidative stress.

Parthanatos is a form of regulated cell death (RCD) that is closely linked to DNA damage, which is a common consequence of oxidative stress due to central nervous trauma, such as spinal cord injury (SCI). The mechanism by which apoptosis-inducing factor (AIF) mediates DNA strand breaks in parthanatos was not clear until the discovery of the nuclease function of MIF. A previous study suggested that observed results may not be reliable if the oxidative stress induced in cells observed under experimental pathological conditions does not accurately replicate the specific pathologies being studied. According to an earlier direct measurement of extracellular oxidative stress in a rat SCI model, post-SCI oxidative stress was approximately the same as exposure to 150 µM H2O2. However, this concentration has been reported as sublethal oxidative stress in other cell types related to senescence, apoptosis, and parthanatos. Using sublethal H2O2 concentrations to induce oxidative stress is equivocal. Also, different cell types have diverse tolerances and responses to oxidative stress, and, therefore, exposure to H2O2. To avoid these limitations, the present study explored the mechanism of neuronal death under this simulated post-SCI oxidative stress and determined the effects of MIF knockdown in parthanatos associated with SCI. Immunofluorescence and flow cytometry were used to reveal typical characteristics of parthanatos that were blocked by PARP-1 inhibitors but not caspase inhibitors. In addition to classic features like PARP-1 and caspase-3 cleavage that were absent, we determined that parthanatos instead of apoptosis played a major role in the cell death caused by oxidative stress following SCI. Flow cytometry analysis of cells transfected by adenovirus with MIF-shRNA then exposed to H2O2 showed a significant decrease in cell death for MIF knockdown cells, even after AIF nuclear translocation. The comet assay also displayed significantly fewer DNA strand breaks after MIF knockdown. This is the first study has verified that MIF knockdown enables to protect neurons from parthanatos under a simulated in vivo oxidative stress following SCI. It suggests that MIF knockdown is a promising therapy to rescue neurons suffering from oxidative stress-induced SCI pathology.

Stromal ColXα1 expression correlates with tumor-infiltrating lymphocytes and predicts adjuvant therapy outcome in ER-positive/HER2-positive breast cancer.

BACKGROUND: The breast cancer microenvironment contributes to tumor progression and response to chemotherapy. Previously, we reported that increased stromal Type X collagen α1 (ColXα1) and low TILs correlated with poor pathologic response to neoadjuvant therapy in estrogen receptor and HER2-positive (ER+/HER2+) breast cancer. Here, we investigate the relationship of ColXα1 and long-term outcome of ER+/HER2+ breast cancer patients in an adjuvant setting. METHODS: A total of 164 cases with at least 5-year follow-up were included. Immunohistochemistry for ColXα1 was performed on whole tumor sections. Associations between ColXα1expression, clinical pathological features, and outcomes were analyzed. RESULTS: ColXα1 expression was directly proportional to the amount of tumor associated stroma (p = 0.024) and inversely proportional to TILs. Increased ColXα1 was significantly associated with shorter disease free survival and overall survival by univariate analysis. In multivariate analysis, OS was lower in ColXα1 expressing (HR = 2.1; 95% CI = 1.2-3.9) tumors of older patients (> = 58 years) (HR = 5.3; 95% CI = 1.7-17) with higher stage (HR = 2.6; 95% CI = 1.3-5.2). Similarly, DFS was lower in ColXα1 expressing (HR = 1.8; 95% CI = 1.6-5.7) tumors of older patients (HR = 3.2; 95% CI = 1.3-7.8) with higher stage (HR = 2.7; 95% CI = 1.6-5.7) and low TILs. In low PR+ tumors, higher ColXα1 expression was associated with poorer prognosis. CONCLUSION: ColXα1 expression is associated with poor disease free survival and overall survival in ER+/HER2+ breast cancer. This study provides further support for the prognostic utility of ColXα1 as a breast cancer associated stromal factor that predicts response to chemotherapy.

Fatty acid-binding protein 1 is preferentially lost in microsatellite instable colorectal carcinomas and is immune modulated via the interferon γ pathway.

Fatty acid-binding protein 1 (FABP1) is an intracellular protein responsible for the transportation of long chain fatty acids. Aside from its functions in lipid metabolism and cellular differentiation, FABP1 also plays a role in inflammation through its interaction with peroxisome proliferator-activated receptors (PPARs). Previously, we compared expression of colonic epithelium genes in a subset of microsatellite instable (MSI) colorectal carcinomas (medullary carcinomas) to normal colonic mucosa and found that FABP1 expression was markedly decreased in the tumors. Further analysis of RNA expression in the colorectal subtypes and The Cancer Genome Atlas data set found that FABP1 expression is decreased in the CMS1 subset of colorectal carcinomas, which is characterized by microsatellite instability. As MSI colorectal carcinomas are known for their robust immune response, we then aimed to link FABP1 to the immune microenvironment of MSI carcinomas. To confirm the gene expression results, we performed immunohistochemical analysis of a cohort of colorectal carcinomas. FABP1 was preferentially lost in MSI carcinomas (123/133, 93%) compared with microsatellite stable carcinomas (240/562, 43%, P<0.0001). In addition, higher numbers of tumor-infiltrating lymphocytes were present in tumors with loss of FABP1 (P<0.0001). Decreased expression of the fatty acid storage and glucose regulator, PPARγ, was associated with the loss of FABP1 (P<0.0001). Colorectal cancer cell lines treated with interferon γ exhibited decreased expression of FABP1. FABP1 expression was partially recovered with the treatment of the cell lines with rosiglitazone, a PPARγ agonist. This study demonstrated that the loss of FABP1 expression is associated with MSI carcinomas and that interferon γ stimulation plays a role in this process via its interaction with PPARγ.

Medullary carcinoma of the colon: a distinct morphology reveals a distinctive immunoregulatory microenvironment.

Medullary carcinoma of the colon is a unique histologic subtype of microsatellite unstable colorectal carcinoma but little is known regarding its tumor-immunoregulatory microenvironment. The aims of this study were to characterize the immune environment of medullary carcinoma and compare it with other microsatellite unstable and microsatellite stable colorectal carcinomas. An initial gene expression microarray analysis of six cases of medullary carcinoma was used to detect potentially differentially expressed genes. We extended this analysis utilizing genomic data from the Cancer Genome Atlas to compare eight cases of medullary carcinoma with other microsatellite unstable and stable carcinomas. Finally, we evaluated expression of key immune pathway proteins and lymphocyte subsets via immunohistochemistry of a large group of medullary carcinomas (n=105) and compared these findings with three other groups: poorly differentiated, microsatellite unstable well-differentiated and microsatellite stable well-differentiated carcinomas. Microarray and the Cancer Genome Atlas data analysis identified significant upregulation of several immunoregulatory genes induced by IFNγ including IDO-1, WARS (tRNA(trp)), GBP1, GBP4, GBP5, PDCD1 (PD-1), and CD274 (PD-L1) in medullary carcinoma compared with other microsatellite unstable and microsatellite stable tumors. By immunohistochemistry, IDO-1 was expressed in 64% of medullary carcinomas compared with 19% (9/47) of poorly differentiated carcinomas, 14% (3/22) of microsatellite unstable, and 7% (2/30) of the microsatellite stable well-differentiated carcinomas (P<0.0001). tRNA(trp) was overexpressed in 81% (84/104) of medullary carcinomas, 19% (9/47) of poorly differentiated, 32% (7/22) of microsatellite unstable, and 3% (1/30) of microsatellite stable well-differentiated carcinomas (P<0.0001). Medullary carcinoma had higher mean CD8+ and PD-L1+ tumor-infiltrating lymphocytes compared with all other groups (P<0.0001). This study demonstrates overexpression of several immunoregulatory genes in microsatellite unstable colorectal carcinomas and that expression of these genes and proteins is more prevalent in the medullary carcinoma subtype, which may be of use both diagnostically and therapeutically.

Identification of stromal ColXα1 and tumor-infiltrating lymphocytes as putative predictive markers of neoadjuvant therapy in estrogen receptor-positive/HER2-positive breast cancer.

BACKGROUND: The influence of the tumor microenvironment and tumor-stromal interactions on the heterogeneity of response within breast cancer subtypes have just begun to be explored. This study focuses on patients with estrogen receptor-positive/human epidermal growth factor receptor 2-positive (ER+/HER2+) breast cancer receiving neoadjuvant chemotherapy and HER2-targeted therapy (NAC+H), and was designed to identify novel predictive biomarkers by combining gene expression analysis and immunohistochemistry with pathologic response. METHODS: We performed gene expression profiling on pre-NAC+H tumor samples from responding (no or minimal residual disease at surgery) and non-responding patients. Gene set enrichment analysis identified potentially relevant pathways, and immunohistochemical staining of pre-treatment biopsies was used to measure protein levels of those pathways, which were correlated with pathologic response in both univariate and multivariate analysis. RESULTS: Increased expression of genes encoding for stromal collagens, including Col10A1, and reduced expression of immune-associated genes, reflecting lower levels of total tumor-infiltrating lymphocytes (TILs), were strongly associated with poor pathologic response. Lower TILs in tumor biopsies correlated with reduced likelihood of achieving an optimal pathologic response, but increased expression of the Col10A1 gene product, colXα1, had greater predictive value than stromal abundance for poor response (OR = 18.9, p = 0.003), and the combination of increased colXα1 expression and low TILs was significantly associated with poor response in multivariate analysis. ROC analysis suggests strong specificity and sensitivity for this combination in predicting treatment response. CONCLUSIONS: Increased expression of stromal colXα1 and low TILs correlate with poor pathologic response in ER+/HER2+ breast tumors. Further studies are needed to confirm their predictive value and impact on long-term outcomes, and to determine whether this collagen exerts a protective effect on the cancer cells or simply reflects other factors within the tumor microenvironment.

[Comparison of effectiveness between unilateral biportal endoscopic decompression and unilateral biportal endoscopic lumbar interbody fusion for degree â degenerative lumbar spondylolisthesis].

Objective: To compare the effectiveness of unilateral biportal endoscopic decompression and unilateral biportal endoscopic lumbar interbody fusion (ULIF) in the treatment of degreeⅠdegenerative lumbar spondylolisthesis (DLS). Methods: A clinical data of 58 patients with degreeⅠDLS who met the selection criteria between October 2021 and October 2022 was retrospectively analyzed. Among them, 28 cases were treated with unilateral biportal endoscopic decompression (decompression group) and 30 cases with ULIF (ULIF group). There was no significant difference between the two groups ( P>0.05) in the gender, age, lesion segment, and preoperative visual analogue scale (VAS) score of low back pain, VAS score of leg pain, Oswestry disability index (ODI), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), disk height (DH), segmental lordosis (SL), and other baseline data. The operation time, postoperative drainage volume, postoperative ambulation time, VAS score of low back pain, VAS score of leg pain, ODI, laboratory examination indexes (CRP, ESR), and imaging parameters (DH, SL) were compared between the two groups. Results: Compared with the ULIF group, the decompression group had shorter operation time, less postoperative drainage, and earlier ambulation ( P<0.05). All incisions healed by first intention, and no complication such as nerve root injury, epidural hematoma, or infection occurred. All patients were followed up 12 months. Laboratory tests showed that ESR and CRP at 3 days after operation in decompression group were not significantly different from those before operation ( P>0.05), while the above indexes in ULIF group significantly increased at 3 days after operation compared to preoperative values ( P<0.05). There were significant differences in the changes of ESR and CRP before and after operation between the two groups ( P<0.05). Except that the VAS score of low back pain at 3 days after operation was not significantly different from that before operation in decompression group ( P>0.05), there were significant differences in VAS score of low back pain and VAS score of leg pain between the two groups at other time points ( P<0.05). The VAS score of low back pain in ULIF group was significantly higher than that in decompression group at 3 days after operation ( P<0.05), and there was no significant difference in VAS score of low back pain and VAS score of leg pain between the two groups at other time points ( P>0.05). The ODI of the two groups significantly improved after operation ( P<0.05), but there was no significant difference between 3 days and 6 months after operation ( P>0.05). There was no significant difference between the two groups at the two time points after operation ( P<0.05). Imaging examination showed that there was no significant difference in DH and SL between pre-operation and 12 months after operation in decompression group ( P>0.05). However, the above two indexes in ULIF group were significantly higher than those before operation ( P<0.05). There were significant differences in the changes of DH and SL before and after operation between the two groups ( P<0.05). Conclusion: Unilateral biportal endoscopic decompression can achieve good effectiveness in the treatment of degree Ⅰ DLS. Compared with ULIF, it can shorten operation time, reduce postoperative drainage volume, promote early ambulation, reduce inflammatory reaction, and accelerate postoperative recovery. ULIF has more advantages in restoring intervertebral DH and SL.

CARMA3 Drives NF-κB Activation and Promotes Intervertebral Disc Degeneration: Involvement of CARMA3-BCL10-MALT1 Signalosome.

Intervertebral disc degeneration (IDD) diseases are common and frequent diseases in orthopedics. The caspase recruitment domain (CARD) and membrane-associated guanylate kinase-like protein 3 (CARMA3) is crucial in the activation of the NF-κB pathway. However, the biological function of CARMA3 in IDD remains unknown. Here, CARMA3 expression was elevated in nucleus pulposus (NP) tissues of IDD rats and nutrient deprivation (ND)-induced NP cells. The main pathological manifestations observed in IDD rats were shrinkage of the NP, reduction of NP cells, fibrosis of NP tissues, and massive reduction of proteoglycans. These changes were accompanied by a decrease in the expression of collagen II and aggrecan, an increase in the expression of the extracellular matrix (ECM) catabolic proteases MMP-3, MMP-13, and metalloprotease with ADAMTS-5, and an increase in the activity of the pro-apoptotic protease caspase-3. The expression of p-IκBαSer32/36 and p-p65Ser536 was also upregulated. However, these effects were reversed with the knockdown of CARMA3. Mechanistically, CARMA3 bound to BCL10 and MALT1 to form a signalosome. Knockdown of CARMA3 reduced the CARMA3-BCL10-MALT1 signalosome-mediated NF-κB activation. CARMA3 activated the NF-κB signaling pathway in a manner that bound to BCL10 and MALT1 to form a signalosome, which affects NP cell damage and is involved in the development of IDD. This supports CARMA3-BCL10-MALT1-NF-κB as a promising targeting axis for the treatment of IDD.

Angiotensin receptor blockers and statins could alleviate atrial fibrosis via regulating platelet-derived growth factor/Rac1/nuclear factor-kappa B Axis.

AIMS: To investigate whether the administration of renin-angiotensin system (RAS) inhibitors and statins could alleviate atrial fibrosis via platelet-derived growth factor (PDGF)/Rac1 /nuclear factor-kappa B (NF-κB) axis. METHODS AND RESULTS: In human left atrium, the degree of atrial fibrosis, as well as the expression levels of PDGF, Rac1 and NF-κB increased 1.5 to 2.9 folds in patients with atrial fibrillation compared to that with sinus rhythm, (P<0.0001). There were strongly positive correlations between angiotensin II (Ang II) or procollagen type III-alpha-1 (COL3A1) with PDGF, Rac1, NF-κB, and among PDGF, Rac1 and NF-κB (all P<0.05). At 3 weeks after the transverse aorta constriction (TAC) operation in rat model and with intervention of irbesartan or/and simvastatin, the collagen volume fraction (CVF) and atrial natriuretic peptide (ANP) values respectively increased 6-folds and 3.5-folds in the TAC group compared to SHAM group (P<0.0001), but these levels decreased by 16% to 63% with following drug intervention (all P<0.0001), the combined treatment was the lowest. Accordingly, the expression levels of PDGF (3-folds), Rac1 (1.6-folds), NF-κB (7-folds) and AngII (12-folds) significantly increased in the TAC group compared to the SHAM group, and these levels were also reduced by 25% to 64% with following drug intervention. The highest reduction could be seen after treatment with irbesartan and simvastatin in combination (all P<0.001).There were strongly positive correlations between AngII or CVF with PDGF, Rac1, NF-κB, and among PDGF, Rac1 and NF-κB (all P<0.05). CONCLUSIONS: Irbesartan or/and simvastatin can improve atrial fibrosis by regulating PDGF/Rac1/NF-κB axis.

Direct fabrication of NbS2 nanoflakes on carbon fibers by atomic layer deposition for ultrasensitive cardiac troponin I detection.

The sensitive detection of cardiac troponin I (cTnI) is of great significance for the early diagnosis of acute myocardial infarction (AMI). Herein, in order to fabricate an electrochemical biosensor for ultrasensitive cTnI detection, atomic layer deposition (ALD) was employed to directly deposit NbS2 nanoflakes (NFs) on carbon fiber paper (CFP). Due to the self-limiting reaction of ALD, NbS2NFs were deposited uniformly and accurately on the surface of carbon fibers by controlling the number of ALD cycles, which ensured ultrasensitive detection. Precise regulation of the nanoscale morphology and electrochemical performance of NbS2 nanoflakes via ALD cycles was observed in depth. Owing to the high surface area and conductivity, an anodic/cathodic current of ∼3.01 mA of NbS2NFs/CFP can be obtained. Subsequently, an electrochemical biosensor based on the excellent performance of NbS2NFs/CFP was fabricated. The ultrasensitive detection of cTnI in a linear range of 1 fM to 0.1 nM with a detection limit of 0.32 fM was achieved.

Co-treatment of diamond-wire-saw silicon kerf and spent automotive catalysts for simultaneous recovery of PGMs, REEs, Zr, and high-purity Si.

Spent automotive catalysts (SACs) and diamond-wire-saw silicon kerf (DWSSK) are classified as hazardous wastes. Currently, the two wastes are treated separately using unrelated approaches. More than two independent approaches are required to recover platinum group metals (PGMs), Zr and rare earth elements (REEs) from SACs, and recover Si from DWSSK, which is time-consuming and laborious. In this study, a new approach was proposed to co-treat the two wastes based on the concept of using waste treats waste: using DWSSK (∼89.85 wt% Si) as a new metal collector to extract PGMs, REEs, and Zr simultaneously from SACs to obtain a Si-VM alloy (VM: valuable metal); meanwhile, using the carrier of SACs to form molten slag to eliminate the main impurity, O, from DWSSK. The largest recovery ratios of Pd, Rh, Zr, Ce, La, and Nd from SACs were 99.50 ± 0.10%, 99.14 ± 0.14 %, 96.19 ± 0.76%, 67.18 ± 4.57%, 61.24 ± 4.93% and 47.65 ± 7.27%, respectively, and the largest removal ratio of O from DWSSK was 99.96%. After smelting, the Si-VM alloy was separated into high-purity Si and VM-containing acid solutions via acid leaching. The leaching ratios of Pd, Rh, Ce, La, Nd, and Zr were 99.78%, 98.15%, 99.93%, ∼100%, 99.76% and 99.98%, respectively. The purity of Si was upgraded from 89.85 wt% (in DWSSK) to 99.98 wt% after acid leaching. The new approach proposed in this study is considered more environmentally friendly and cost-effective than the regular approaches that treat the two wastes separately.

Utility of Wnt family member 9b (Wnt9b) immunohistochemistry in the cytologic diagnosis of metastatic breast carcinoma.

Wnt family member 9b (Wnt9b) has been demonstrated as a valuable marker for breast cancer diagnosis in surgical pathology. In this study, we examined the utility of Wnt9b in diagnosing metastatic breast carcinoma in cytology samples. Cell blocks from fine needle aspirations (FNA) and fluid specimens of 96 metastatic breast carcinomas and 123 primary and metastatic non-breast neoplasms from various organ systems were evaluated by Wnt9b and GATA3 immunohistochemistry (IHC). Wnt9b and GATA3 were positive in 81.3% and 92.7% of metastatic breast carcinomas, respectively. Conversely, 93.5% and 90.0% of non-breast, non-urothelial carcinomas were negative for Wnt9b and GATA3, respectively. Wnt9b expression was positive in rare gastrointestinal, gynecological, lung, pancreas, and salivary gland tumors. All twenty-eight urothelial carcinomas were negative for Wnt9b, while twenty-six (92.9%) were positive for GATA3. Wnt9b was slightly less sensitive but more specific than GATA3 in diagnosing metastatic breast cancer in cytology samples. Particularly, Wnt9b shows higher specificity in differentiating breast and urothelial primaries. The combined use of Wnt9b and GATA3 may increase diagnostic accuracy.

Triple-negative Breast Carcinoma With Apocrine and Histiocytoid Features: A Clinicopathologic and Molecular Study of 18 Cases.

Triple-negative breast cancer (TNBC) is a heterogenous group of tumors. Most TNBCs are high-grade aggressive tumors, but a minority of TNBCs are not high grade, with relatively indolent behavior and specific morphologic and molecular features. We performed a clinicopathologic and molecular assessment of 18 non-high-grade TNBCs with apocrine and/or histiocytoid features. All were grade I or II with low Ki-67 (≤20%). Thirteen (72%) showed apocrine features, and 5 (28%) showed histiocytoid and lobular features. In all, 17/18 expressed the androgen receptor, and 13/13 expressed gross cystic disease fluid protein 15. Four (22.2%) patients were treated with neoadjuvant chemotherapy, but none achieved a pathologic complete response. In all, 2/18 patients (11%) had lymph node metastasis at the time of surgery. None of the cases had a recurrence or disease-specific death, with an average follow-up time of 38 months. Thirteen cases were profiled by targeted capture-based next-generation DNA sequencing. Genomic alterations (GAs) were most significant for PI3K-PKB/Akt pathway (69%) genes, including PIK3R1 (23%), PIK3CA (38%), and PTEN (23%), and RTK-RAS pathway (62%) including FGFR4 (46%) and ERBB2 (15%). TP53 GA was seen in only 31% of patients. Our findings support those on high-grade TNBCs with apocrine and/or histiocytoid features as a clinicopathologic and genetically distinct subgroup of TNBC. They can be defined by features including tubule formation, rare mitosis, low Ki-67 (≤20%), triple-negative status, expression of androgen receptor and/or gross cystic disease fluid protein 15, and GA in the PI3K-PKB/Akt and/or RTK-RAS pathway. These tumors are not sensitive to chemotherapy but have favorable clinical behavior. Tumor subtype definitions are the first step to implementing future trial designs to select these patients.

Hypolipidemic agent Z-guggulsterone: metabolism interplays with induction of carboxylesterase and bile salt export pump.

Z-Guggulsterone is a major ingredient in the Indian traditional hypolipidemic remedy guggul. A study in mice has established that its hypolipidemic effect involves the farnesoid X receptor (FXR), presumably by acting as an antagonist of this receptor. It is generally assumed that the antagonism leads to induction of cytochrome P450 7A1 (CYP7A1), the rate-limiting enzyme converting free cholesterol to bile acids. In this study, we tested whether Z-guggulsterone indeed induces human CYP7A1. In addition, the expression of cholesteryl ester hydrolase CES1 and bile salt export pump (BSEP) was monitored. Contrary to the general assumption, Z-guggulsterone did not induce CYP7A1. Instead, this phytosterol significantly induced CES1 and BSEP through transactivation. Z-Guggulsterone underwent metabolism by CYP3A4, and the metabolites greatly increased the induction potency on BSEP but not on CES1. BSEP induction favors cholesterol elimination, whereas CES1 involves both elimination and retention (probably when excessively induced). Interestingly, clinical trials reported the hypolipidemic response rates from 18% to 80% and showed that higher dosages actually increased VLDL cholesterol. Our findings predict that better hypolipidemic outcomes likely occur in individuals who have a relatively higher capacity of metabolizing Z-guggulsterone with moderate CES1 induction, a scenario possibly achieved by lowering the dosing regimens.

Surge in expression of carboxylesterase 1 during the post-neonatal stage enables a rapid gain of the capacity to activate the anti-influenza prodrug oseltamivir.

BACKGROUND: Oseltamivir, a widely used anti-influenza drug, is hydrolytically activated by carboxylesterase 1 (CES1). The expression of this carboxylesterase is developmentally regulated. This study was performed to determine when after birth infants acquire competence of activating this prodrug. METHODS: Liver tissue samples were collected and divided into 5 age groups: group 1 (1-31 d old), group 2 (35-70 d old), group 3 (89-119 d old), group 4 (123-198 d old), and group 5 (>18 years of age). These samples were analyzed for oseltamivir hydrolysis and CES1 expression. RESULTS: Liver samples in group 1 expressed the lowest level of CES1 with the lowest hydrolytic activity toward oseltamivir. A 4-7-fold increase between groups 1 and 2 (1-31 vs 35-70 d of age) was detected in the hydrolysis and expression analyses, respectively. Liver samples in the other 3 pediatric groups (35-198 d of age) exhibited similar expression and hydrolysis levels. Overall, liver samples in group 1 had CES1 expression and hydrolysis levels that were 10% of those of adults, whereas liver samples in the other 3 pediatric groups had levels that were ∼50% of adult levels. CONCLUSIONS: The post-neonatal surge in CES1 expression ensures the hydrolytic capacity to be gained rapidly after birth in infants, but the larger variability during this period suggests that caution should be exercised on the extrapolated dosing regimens of ester drugs from other age groups.

Diagnosis and Treatment of Cervical Spondylotic Radiculopathy Using Selective Nerve Root Block (SNRB): Where are We Now?

Cervical spondylotic radiculopathy (CSR) is one of the most common degenerative diseases of the spine that is commonly treated with surgery. The primary goal of surgery is to relieve symptoms through decompression or relieving pressure on compressed cervical nerves. Nevertheless, cutaneous pain distribution is not always predictable, making accurate diagnosis challenging and increasing the likelihood of inadequate surgical outcomes. With the widespread application of minimally invasive surgical techniques, the requirement for precise preoperative localization of the affected segments has become critical, especially when treating patients with multi-segmental CSR. Recently, the preoperative use of a selective nerve root block (SNRB) to localize the specific nerve roots involved in CSR has increased. However, few reviews discuss the currently used block approaches, risk factors, and other aspects of concern voiced by surgeons carrying out SNRB. This review summarized the main cervical SNRB approaches currently used clinically and the relevant technical details. Methods that can be used to decrease risk during cervical SNRB procedures, including choice of steroids, vessel avoidance, guidance with radiographs or ultra-sound, contrast agent usage, and other concerns, also are discussed. We concluded that a comprehensive understanding of the current techniques used for cervical SNRB would allow surgeons to perform cervical SNRB more safely.

Clinical efficacy and imaging outcomes of unilateral biportal endoscopy with unilateral laminotomy for bilateral decompression in the treatment of severe lumbar spinal stenosis.

Objective: To investigate the clinical efficacy and imaging outcomes of unilateral biportal endoscopy (UBE) with unilateral laminotomy for bilateral decompression (ULBD) in the treatment of severe lumbar spinal stenosis (SLSS). Methods: We retrospectively analyzed 50 patients with SLSS treated with UBE-ULBD from October 2018 to March 2021. Visual analog scale (VAS) for back and legs pain, Oswestry disability index (ODI), modified Macnab criteria, complications, hospital stay, preoperative and postoperative dural sac cross-sectional area (DSCA) and Schizas grade, mean angle of facetectomy and osseous lateral recess decompression rate were examined. Results: The mean follow-up period was 10.7 months. The mean hospital stay was 2.76 ± 1.02 days. At the final follow-up, VAS for back pain and legs pain decreased from 7.22 ± 0.95 to 1.26 ± 0.44 and from 7.88 ± 0.69 to 1.18 ± 0.39, respectively; ODI decreased from 69.88 ± 6.32% to 14.96 ± 2.75%. According to the modified Macnab criteria, the results were excellent in 24 (48%), good in 22 (44%), and fair in 4 (8%). Excellent or good results (a satisfactory outcome) were obtained in 92% of the patients. There were 2 cases of complications of dural sac tear. The postoperative DSCA was significantly enlarged compared with that before surgery, from 44.74 ± 9.85 to 126.86 ± 14.81 mm2. According to Schizas grade, the stenosis grade changes from preoperative grade C in 16 cases, grade D in 34 cases, to postoperative grade A in 40 cases, and grade B in 10 cases. The mean angle of facetectomy of the ipsilateral facet joint was 70.87 ± 5.68 ∘ , contralateral was 65.07 ± 4.98 ∘ . The decompression rate was 70.81 ± 4.43% (ipsilateral side) and 71.22 ± 3.68% (contralateral). Conclusions: UBE-ULBD has a good clinical effect in the treatment of SLSS, and has achieved satisfactory results in spinal canal enlargement, undercutting of facet joints, and decompression effect. It is a safe and effective surgical for SLSS.

Utility of secretagogin as a marker for the diagnosis of lung neuroendocrine carcinoma.

Small-cell lung cancers (SCLC) and large-cell neuroendocrine carcinomas (LCNEC) are two types of high-grade pulmonary neuroendocrine carcinomas (NECs). Diagnostic neuroendocrine markers commonly include synaptophysin, chromogranin A, CD56, and insulinoma-associated protein 1 (INSM1). In this study, the utility of secretagogin (SCGN) was examined in the context of pulmonary NEC diagnosis. The study included 71 pulmonary NEC cases (18 SCLCs, 13 combined-SCLCs, 23 LCNECs, and 17 combined-LCNECs). Immunohistochemical stains of SCGN, synaptophysin, chromogranin A, CD56, and INSM1 were performed on whole tumor sections. The stains were evaluated based on combined staining intensity and the proportion of positive tumor cells. At least mild staining intensity in at least 1% of the cells was considered positive. Bioinformatic studies showed specific SCGN expression in neuroendocrine cells and NECs. SCGN showed diffuse nuclear and cytoplasmic staining in NECs with intra-tumoral heterogeneity. The non-neuroendocrine components were negative. The sensitivity of SCGN was no better than the other established neuroendocrine markers based on all NECs combined or LCNECs/c-LCNECs only. However, the sensitivity of SCGN (71%) was higher than chromogranin A (68%) for SCLCs/c-SCLCs only. The average proportion of SCGN positive tumor cells was 8% higher than chromogranin A (22% versus 14%, P = 0.0332) in all NECs and 18% higher for SCLC and c-SCLC cases only (32% versus 13%, P = 0.0054). The above data showed that SCGN could be used as a supplemental neuroendocrine marker to diagnose SCLC.