A novel necroptosis-related model predicts the prognosis and immunocompetence of head and neck squamous cell carcinoma.

Head and neck squamous cell carcinoma (HNSCC) is a disease characterized by profound immunosuppression and the prognosis of HNSCC patients remains poor. Necroptosis is a programmed lytic cell-death mechanism that can promote tumor growth, angiogenesis, invasion and metastasis. The differentially expressed NRGs were screened by using the LIMMA package of R software (version 4.1.2) and the prognosis-related NRGs were obtained by COX regression analysis. We separated patients into high- and low-risk groups via the prognostic model consisting of those NRGs. The receiver operating characteristic (ROC) curve and Kaplan-Meier survival curves were used to validate the prognostic model. By bioinformatics analysis, the prognostic risk and immunocompetent models were constructed. We reevaluate the prognostic model based on the GES27020 data sets, clinicopathological variables and chemotherapeutic efficacy. Individuals in the high-risk group had much shorter overall survival (OS) times than their counterparts. Compared with clinicopathological variables, the risk model has a higher diagnostic efficiency, with the area under the ROC being 0.699. Decision Curve Analysis (DCA) showed the prognostic model-based risk score was the superior prognostic factor compared to additional indicators. Furthermore, the high- and low-risk groups had differences in immune cell infiltration and immune functions. And the CCK-8 showed that small molecular drugs could inhibit HNSCC cell proliferation in vitro. We have constructed a new necroptosis-related model, which can be used to predict the prognosis and immunocompetence of HNSCC patients and provide a theoretical basis for the study of necroptosis in the clinical prognosis of HNSCC.

An oligosaccharide marker for rapid authentication of edible bird's nest.

Edible bird's nest (EBN) is a popular and expensive food material. The limited supply and great demand result in the use of adulterants. The authenticity concern is raised due to the lack of appropriate quality markers. Herein, this study aims to provide a specific oligosaccharide marker for rapid EBN authentication. Comparing the benzocaine (ABEE)-labeled saccharide profiles of multiple batches of EBN and adulterants indicates seven unique EBN oligosaccharides. The most abundant one, named BNM001, was selected as a marker and characterized to be Neu5Ac (2-3) Gal by MS and NMR spectra. This new oligosaccharide marker enables a rapid authentication of EBN within 10 min. ABEE labelling of this marker further upgraded the accuracy and sensitivity of the LC-qTOF-MS quantitative analysis. The relative marker content was associated with the quality of EBN products. These results suggest a specific and efficient quality marker for rapid authentication of EBN and related products.

Elevated serum levels of diamine oxidase, D-lactate and lipopolysaccharides are associated with metabolic-associated fatty liver disease.

BACKGROUND: Studies have suggested an association between metabolic-associated fatty liver disease (MAFLD) and intestinal barrier function. The present study aims to investigate the association between MAFLD and intestinal barrier impairment in humans and identify potential risk factors for MAFLD. METHODS: A total of 491 patients were retrospectively enrolled in this study. The serum levels of diamine oxidase, D-lactate and lipopolysaccharide were measured to evaluate intestinal barrier integrity in patients with and without MAFLD. Binary logistic regression and correlational analyses were conducted to verify the association between MAFLD and serum levels of intestinal barrier biomarkers. RESULTS: We enrolled 294 patients with MAFLD and 197 patients without MAFLD in this study. Patients with MAFLD had higher serum levels of diamine oxidase, D-lactate and lipopolysaccharide (P < 0.001) than those without MAFLD. Multivariate logistic regression analyses showed that BMI [odds ratio (OR) 1.324; P < 0.001], triglycerides (OR 2.649; P = 0.002), nonesterified fatty acids (OR 1.002; P = 0.011), diamine oxidase (OR 1.149; P = 0.011) and D-lactate (OR 1.221; P < 0.001) were independent risk factors for MAFLD. Additionally, serum levels of diamine oxidase and D-lactate increase as liver steatosis became more severe. MAFLD patients with ≥2 metabolic abnormalities had higher serum levels of lipopolysaccharide (P = 0.034). CONCLUSIONS: MAFLD is associated with intestinal barrier impairment. Diamine oxidase and D-lactate are potential predictors of MAFLD, and their serum levels are related to liver steatosis. Intestinal barrier impairment is related to metabolic disorders in patients with MAFLD.

The effects of local infiltration anesthesia and femoral nerve block analgesia after total knee arthroplasty: a systematic review and meta-analysis.

Background: Local infiltration anesthesia (LIA) and femoral nerve block (FNB) are commonly used analgesia methods after total knee arthroplasty (TKA). However, there is no definitive conclusion about which of these two analgesia modes is superior. Therefore, this study aimed to systematically evaluate the analgesic effects of LIA and FNB after TKA. Methods: We used the terms "total knee replacement, knee replacement, total knee arthroplasty, knee arthroplasty, local infiltration analgesia, periarticular infiltration, periarticular injection, intra-articular infiltration, intra-articular injection, peripheral nerve block, femoral nerve block" to search the PubMed, Cochrane Central Register of Controlled Trials, Embase, Web of Science, China National Knowledge Infrastructure (CNKI), Wanfang, and Weipu databases. The search period was set from the date of establishment of the database to September 2021. The Cochrane risk of bias tool was used to evaluate the quality of the included studies, and network meta-analysis was performed using Stata14.0 and RevMan 5.30 software. Results: Nine articles were included for analysis. The results of meta-analysis showed that compared with LIA and FNB, the difference in opioid use [mean difference (MD) -4.35, 95% confidence interval (CI): -7.26 to -1.45] was statistically significant. However, there was no significant difference between the static visual analogue score at 24 hours postoperatively (MD 0.20, 95% CI: -0.91 to 1.31), the visual analogue score for exercise visual analogy at 24 hours after surgery (MD 0.10, 95% CI: -0.12 to 0.32), and the length of hospital stay (MD 0.05, 95% CI: -0.40 to 0.50). Discussion: LIA and FNB have similar effects on pain relief after TKA, but LIA can reduce the use of analgesic drugs and is easy to operate. Therefore, LIA can be used as the priority analgesic method for patients with TKA. However, multi-center, large-sample, high-quality, randomized controlled trials are still needed for further verification.

Open-Air Dual-Diamination of Aromatic Aldehydes: Direct Synthesis of Azolo-Fused 1,3,5-Triazines Facilitated by Ammonium Iodide.

A new and practical protocol for the synthesis of medicinally privileged azolo[1,3,5]triazines by simply heating under air has been presented. The in situ generated N-azolo amidines from commercially available aromatic aldehydes and 3-aminoazoles with ammonium iodide undergo the second diamination to accomplish the [3 + 1 + 1 + 1] heteroannulation reaction. This convenient process is appreciated by high efficiency, broad substrate scope, gram-scale synthesis, and operational simplicity under reagent-free conditions.

A specific and bioactive polysaccharide marker for Cordyceps.

Cordyceps is one of the most expensive and widely used functional foods. But the authenticity is still a concern due to the lack of appropriate markers. By targeting polysaccharides, this study aimed to develop a specific, and bioactive marker for Cordyceps. Firstly, the results of screening tests of 250 samples by examining both genetic markers and polysaccharide profile showed that a unique polysaccharide fraction (named CCP) was particular to the caterpillar parts. Its potential as a marker was further demonstrated by its ability to induce NO and cytokine production in RAW 264.7 cells. CCP was characterized to be an α-1,4-glucan with a branch at C-6 by the conventional structure analyzing and de novo oligosaccharides sequencing. The content of CCP was closely correlated to the traditional classification criteria. Generally, CCP was a marker that simultaneously enables qualitative and quantitative analysis of Cordyceps.

An oligosaccharide-marker approach to quantify specific polysaccharides in herbal formula by LC-qTOF-MS: Danggui Buxue Tang, a case study.

Polysaccharides have broad bioactivities and are major components of water decoction of herb formulae. However, the quality control of polysaccharides remains a challenge. Oligosaccharide-fragment approach has been considered in elucidating chemical structures of polysaccharides, but never been used for quantitation. Using reference chemicals and a real sample Danggui Buxue Tang (DBT) in this study, an oligosaccharide-marker approach was established to quantify specific polysaccharides. Firstly, linear relationships between parent polysaccharides and hydrolysis-produced daughter oligosaccharides were verified using reference polysaccharides. Then in case of DBT, two fluorescence-labeled oligosaccharides with high specificity to individual parent polysaccharides were selected as markers. They were easily isolated and identified. Their potential in quantification of parent polysaccharides were satisfactorily validated in terms of linearity (r≥0.99), repeatability (RSD ≤ 8.4 %), and spike recovery (≥80 %). This method could be a promising approach for quality assessment of polysaccharides in herbal formulae.

Oligosaccharide-marker approach for qualitative and quantitative analysis of specific polysaccharide in herb formula by ultra-high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry: Dendrobium officinale, a case study.

Qualitative and quantitative analysis of polysaccharides in herb formula remain challenge due to the limited choices of analytical methods concerning the intrinsic characteristics of large molecular mass. Herein, an oligosaccharide-marker approach was newly developed for quality assessment of polysaccharides in herbal materials, using Dendrobium officinale as a case study. This method involved partial acid hydrolysis of D. officinale polysaccharide (DOP) followed by p-aminobenzoic ethyl ester (ABEE) derivatization. Two ABEE-labeled oligosaccharides namely, Te-Man-ABEE and Pen-Man-ABEE, were selected as chemical markers due to their high specificity in herb formula. The linear relationship between the content of these two markers and the content of DOP was then successfully established respectively. The linear relationship was further transformed to that between peak area of chemical markers and DOP content so that chemical markers were not necessary to be isolated for analysis. This linear relationship was systemically validated in terms of precision and accuracy. The results showed that these two oligosaccharide-markers presented a good linear relationship with DOP (R2 ≥ 0.997) in the range of 0.68-16.02 µg. These markers also demonstrated satisfactory precision (RSD < 7.0%), and recovery (91.41%-118.30%) in real sample determination. Additionally, there was no significant difference between the results given by the two chemical markers as the RSD values were not more than 7.0%. While concerning the results given by the oligosaccharide-markers and the previously-published polysaccharide marker, the RSD value was not more than 6.4%. These suggest that the oligosaccharide-marker approach is a simple, quick, and reliable method to qualitatively and quantitatively determine of specific polysaccharide in herb formula.

Asymmetric α-alkylation of cyclic ß-keto esters and ß-keto amides by phase-transfer catalysis.

Without employing any transition metal, a highly enantioselective α-alkylation of cyclic ß-keto esters and ß-keto amides has been realized by phase-transfer catalysis. This improved procedure is applicable to different kinds of halides with cinchona derivatives and gives the corresponding products in excellent enantiopurities (up to 98% ee) and good yields (up to 98%). Moreover, the reaction was scalable and the phase-transfer catalyst was recyclable. This provided an alternative and competitive method to the asymmetric α-alkylation of ß-dicarbonyl compounds.

I2-Triggered N-O cleavage of ketoxime acetates for the synthesis of 3-(4-pyridyl)indoles.

A facile and complementary [3 + 2 + 1] annulation of aryl ketoxime acetates and 3-formylindoles to give pyridine derivatives is reported. The condensation reaction demonstrated that I2 was capable of triggering N-O bond cleavage of ketoxime acetates to generate iminyl radicals via a single electron transfer pathway. This direct and operationally simple protocol provides a fundamental platform to synthesize 3-(4-pyridyl)indoles with high functional group compatibility and high regioselectivity.

Comprehensive comparison of polysaccharides from Ganoderma lucidum and G. sinense: chemical, antitumor, immunomodulating and gut-microbiota modulatory properties.

Both Ganoderma lucidum (GL) and G. sinense (GS) are used as Lingzhi in China. Their functions are assumed to mainly derive from triterpenes and polysaccharides; however, the two species have very different triterpenes profiles, if this was the case, then the bioactivity of these two species should differ. Instead, could the polysaccharides be similar, contributing to the shared therapeutic basis? In this study, two main polysaccharide fractions from different batches of GL and GS were systematically compared by a series of chemical and biological experiments. The results showed that the polysaccharides from two species shared the same structural features in terms of mono-/oligo-saccharide profiles, molecular size, sugar linkages, and IR/NMR spectra. In addition, these polysaccharides showed similar tumor-suppressive activity in mice. Further study on RAW264.7 cells indicated that these polysaccharides exhibited similar inducing effects to macrophages, as evaluated in the phagocytosis function, NO/cytokines production, inhibition against the viability and migration of cancer cells. Mechanistic investigation revealed the identical activation via TLR-4 related MAPK/NF-κB signaling pathway and gut-microbiota modulatory effects. In summary, GL and GS polysaccharides presented similar chemical features, antitumor/immunomodulating activities and mechanism; this establishes polysaccharides as the active principles and supports the official use of both species as Lingzhi.

Alterations in splenic function and gene expression in mice with depressive-like behavior induced by exposure to corticosterone.

Depressed patients present with increased cortisol levels and attenuated immune responses. However, little is known about the association between depression and the spleen, as this is the largest peripheral immune organ. In this study, we examined alterations in splenic function and gene expression in mice with depressive-like behavior, well as the expression of certain proteins in related pathways. A mouse model of depression was established with the use of corticosterone. Splenic function and histopathology were assessed using Wright and H&E staining. The Agilent Whole Mouse Genome Oligo Microarray containing >41,174 transcript probes was used to measure the levels of gene-expression in the spleens from control and model mice, and the levels of certain proteins associated with depression were measured by western blot analysis in the brain and spleen separately. We found that splenic function and immunity in the mice with depressive-like behavior were markedly impaired. A total of 53 genes exhibited a differential response in the mice with depressive-like behavior, 11 of which were more notable, including collagen, type VI, α5 (Col6a5), immunoglobulin superfamily, member 11 (Igsf11), D site albumin promoter binding protein (Dbp), tachykinin 2 (Tac2) and γ-aminobutyric acid B receptor 2 (Gabbr2). Pathway analysis revealed that the amino acid biosynthesis and the clock gene pathways were more meaningful among these genes. The levels of GABBR2, DBP and substance P (SP; encoded by the Tac2 gene) related proteins in the brain were markedly downregulated, and similar results were observed in the spleen. The anti-depressant, fluoxetine, reversed the changes in the levels of these proteins. The findings of our study regarding changes occurring in the spleen during depression may indirectly elucidate and shed light into the pathogenesis of depression and depressive-like behavior.

A Novel Missense Mutation 224G>T (R75M) in SRY Coding Region Interferes with Nuclear Import and Results in 46, XY Complete Gonadal Dysgenesis.

SRY-mutation-caused sex reversal is a rare disease and mostly associated with a de novo mutation since the patients with defective SRY is infertile. There are many reports about SRY-mutation associated 46, XY ovarian disorder of sex development (DSD), but few described their molecular mechanism. Here we report a de novo mutation 224G>T (R75M) in SRY associated with a phenotypic female, 46, XY karyotype and dysgerminoma. The wild and mutated SRY were cloned into recombinant plasmid and expressed in cells in vitro, the result showed the mutated SRY is greatly accumulated in cytoplasm while the wild type SRY is mostly localized in nucleus. To make sure no other genes were involved, we performed the trio-based whole exome sequencing using the DNA samples from the proband and the parents, and no mutations were identified especially in DHH, NR0B1, NR5A1, SOX9 and MAP3K1, indicating the de novo mutation in SRY is the single defect responsible for the female sex reversal. We also used bioinformatics simulation analysis to predict impact of the mutation on SRY function, and find the R75 in wild type SRY can form a hydrogen bond with serine at 91 (S91) that make the SRY protein well fit into the minor groove of target DNA, while the M75 in the mutated SRY can't. Finally, we reviewed SRY mutations based on the available references and analyzed the mutation distribution patterns according to density and continuity, which may be useful for further study of the SRY structure, function, and its relatedness with DSD.

Antidepressant-like and anti-oxidative efficacy of Campsis grandiflora flower.

OBJECTIVES: Our study aimed to investigate the antidepressant-like effect of ethyl acetate extract of the flowers of Campsis grandiflora (EFCG) in a mice model of chronic unpredictable mild stress (CUMS). METHODS: HPLC-Q-TOF-MS was used to identify the chemical constituents of EFCG. The DPPH assay and ABTS radical-scavenging assay were performed to measure the antioxidant properties. The protective properties of EFCG against H2 O2 -induced oxidative damage were analysed in PC12 cells. The changes of behaviour profiles were investigated by using open-field test, sucrose preference test, forced swimming test (FST) and tail suspension test (TST). Brain tissue samples of mice were collected, and antioxidative measure levels were measured. KEY FINDINGS: The result showed that EFCG had the most active anti-oxidative effect and the protective effect against H2 O2 oxidative injury in PC12 cells. Treatment with the EFCG significantly reduced the depressant-like severity and immobility period as compared with untreated CUMS mice in FST and TST. Moreover, EFCG significantly elevated the contents of superoxide dismutase, Glutathione Peroxidase and decreased the contents of Malonaldehyde (MDA) in mice brain. CONCLUSIONS: Our study found first the antidepressant activity of the EFCG. The results suggested the therapeutic potential of EFCG for depressive disorder.

Comparative evaluation of tigecycline susceptibility testing methods for Acinetobacter baumannii and Enterobacteriaceae.

The objectives of this study were to recommend suitable susceptibility testing methods for tigecycline for clinical laboratory use and to evaluate differences in tigecycline susceptibility between carbapenem-susceptible and carbapenem-non-susceptible isolates. Broth microdilution (BMD) was used as the reference method to evaluate MIC Test Strip (MTS), agar dilution, VITEK® 2 and disk diffusion testing methods for tigecycline against Acinetobacter baumannii and Enterobacteriaceae. MIC50/90 values (minimum inhibitory concentrations for 50% and 90% of the isolates, respectively) of A. baumannii and Enterobacteriaceae were, respectively, 2/4µg/mL and 0.5/4µg/mL by BMD, 1.5/3µg/mL and 0.5/3µg/mL by MTS, 2/4µg/mL and 1/8µg/mL by agar dilution and 2/4µg/mL and 2/8µg/mL by VITEK® 2. Essential agreement for A. baumannii/Enterobacteriaceae detected by MTS, agar dilution and VITEK® 2 methods was 96.0/97.3%, 98.0/97.3% and 94.0/63.9%, respectively. Categorical agreement for A. baumannii/Enterobacteriaceae detected by MTS, agar dilution, VITEK® 2 and disk diffusion methods was 90.0/91.8%, 72.0/93.7%, 62.0/86.5% and 72.0/81.2%, respectively. No very major errors were found for all isolates by the four methods evaluated. Major error rates were produced by VITEK® 2 (for Enterobacteriaceae) and by disk diffusion (for A. baumannii and Enterobacteriaceae). Tigecycline susceptibility of carbapenem-susceptible and carbapenem-non-susceptible isolates was 85.2% and 83.6% (χ2=0.15, P>0.05) using the reference method. In conclusion, in this study MTS showed the best correlation with BMD for tigecycline MICs. Based on in vitro testing, tigecycline can be considered an equally useful choice for infections caused by carbapenem-susceptible and carbapenem-non-susceptible isolates.

[Mutation analysis of EXT2 gene in a family with hereditary multiple exostosis].

OBJECTIVE: To investigate EXT1 and EXT2 genes mutations in a family with hereditary multiple osteochondromas (HME). METHODS: A four-generation family with HME from Linyi city of Shandong Province was studied. There were 6 affected individuals among the 17 family members. Physical examination and radiographical evaluations were carried out for all family members. Genomic DNA was extracted from peripheral venous blood and the samples were subjected to mutation screening by PCR of the coding regions of EXT1 and EXT2 genes. RESULTS: The family has featured an autosomal dominant inheritance pattern. Sequencing of the EXT1 and EXT2 genes suggested the causative gene in this family was in linkage with the second exon of EXT2. A c.244delG mutation was detected, which has resulted in a frameshift mutation p.Asp81IlefsX30. The mutation was found in all of the 6 affected individuals but not in normal family members. And the mutation has co-segregated with the phenotype. CONCLUSION: The mutation c.244delG in the EXT2 gene is the probably the cause of the disease in this family.

catena-Poly[[bis-(dicyanamido-κN (1))cobalt(II)]bis-{µ-1,2-bis-[(1,2,4-triazol-1-yl)meth-yl]benzene-κ(2) N (4):N (4')}].

In the title complex, [Co(C2N3)2(C12H12N6)2] n the Co(II) atom lies on a centre of symmetry and displays a slightly distorted octa-hedral coordination geometry. The 1,2-bis-[(1,2,4-triazol-1-yl)meth-yl]benzene ligands link adjacent metal atoms into polymeric chains parallel to the c axis, forming centrosymmetric 26-membered metallamacrocycles. The conformation of the metallamacrocycles is stabilized by pairs of C-H⋯N hydrogen bonds. The dihedral angles formed by the planes of the triazole rings with those of the benzene ring are 79.4 (2) and 79.1 (2)°. In the crystal, the chains inter-act through C-H⋯N hydrogen bonds, forming a three-dimensional network.

Mapping breakpoints of a familial chromosome insertion (18,7) (q22.1; q36.2q21.11) to DPP6 and CACNA2D1 genes in an azoospermic male.

It is widely accepted that the incidence of chromosomal aberration is 10-15.2% in the azoospermic male; however, the exact genetic damages are currently unknown for more than 40% of azoospermia. To elucidate the causative gene defects, we used the next generation sequencing (NGS) to map the breakpoints of a chromosome insertion from an azoospermic male who carries a balanced, maternally inherited karyotype 46, XY, inv ins (18,7) (q22.1; q36.2q21.11). The analysis revealed that the breakage in chromosome 7 disrupts two genes, dipeptidyl aminopeptidase-like protein 6 (DPP6) and contactin-associated protein-like 2 (CACNA2D1), the former participates in regulation of voltage-gated potassium channels, and the latter is one of the components in voltage-gated calcium channels. The deletion and duplication were not identified equal or beyond 100 kb, but 4 homologous DNA elements were verified proximal to the breakpoints. One of the proband's sisters inherited the same aberrant karyotype and experienced recurrent miscarriages and consecutive fetus death, while in contrast, another sister with a normal karyotype experienced normal labor and gave birth to healthy babies. The insertional translocation is confirmed with FISH and the Y-chromosome microdeletions were excluded by genetic testing. This is the first report describing chromosome insertion inv ins (18,7) and attributes DPP6 and CACNA2D1 to azoospermia.