Adulteration Detection and Quantification in Olive Oil Using Excitation-Emission Matrix Fluorescence Spectroscopy and Chemometrics.

This research investigates the use of excitation-emission matrix fluorescence (EEMF) in conjunction with chemometric models to rapidly identify and quantify adulteration in olive oil, a critical concern where sample availability is limited. Adulteration is simulated by blending soybean, peanut, and linseed oils into olive oil, creating diverse adulterated samples. Principal component analysis (PCA) was applied to the EEMF spectral data as an initial exploratory measure to cluster and differentiate adulterated samples. Spatial clustering enabled vivid visualization of the variations and trends in the spectra. The novel application of parallel factor analysis (PARAFAC) for data decomposition in this paper focuses on unraveling correlations between the decomposed components and the actual adulterated components, which offers a novel perspective for accurately quantifying adulteration levels. Additionally, a comparative analysis was conducted between the PCA and PARAFAC methodologies. Our study not only unveils a new avenue for the quantitative analysis of adulterants in olive oil through spectral detection but also highlights the potential for applying these insights in practical, real-world scenarios, thereby enhancing detection capabilities for various edible oil samples. This promises to improve the detection of adulteration across a range of edible oil samples, offering significant contributions to food safety and quality assurance.

Clinical analysis of seven cases of hemophagocytic lymphohistiocytosis secondary to cutaneous T-cell lymphoma / 中华皮肤科杂志

Objective:To investigate clinical and laboratory characteristics of secondary hemophagocytic lymphohistiocytosis (sHLH) associated with secondary cutaneous T-cell lymphoma (CTCL) .Methods:CTCL patients with clinically suspected sHLH were collected from Department of Hematology, Wuhan No.1 Hospital from January 2016 to October 2021, and were evaluated according to the HLH-2004 diagnostic criteria and HScore.Results:Seven CTCL patients were confirmedly diagnosed with sHLH, including 2 with primary cutaneous γδT-cell lymphoma (PC-GDTCL) , 3 with cutaneous extranodal natural killer/T-cell lymphoma (C-ENKTCL) , and 2 with primary cutaneous anaplastic large cell lymphoma (PC-ALCL) . All the 7 patients received chemotherapy, but 6 died finally, and the median overall survival duration was 26.5 days (range: 14 - 60 days) after the confirmed diagnosis of CTCL complicated by sHLH. HLH-related gene mutations, which were located in the PRF1 and LYST genes, were identified in 2 patients; lymphoma-related gene mutations were identified in the KRAS and KMT2D genes in 1 PC-GDTCL patient,and in the JAK3 and SAMHD1 genes in another PC-GDTCL patient.Conclusions:CTCL complicated by sHLH usually progresses rapidly, so early diagnosis and treatment are needed. Bone marrow biopsy and mutation screening of lymphoma- and HLH-related genes at initial diagnosis and during disease progression may facilitate early diagnosis.

Role of S1PR1 in dorsal root ganglion in remifentanil-induced hyperalgesia in rats with incisional pain / 中华麻醉学杂志

Objective:To evaluate the role of sphingosine-1-phospho-1 receptor(S1PR1)in the dorsal root ganglion in remifentanil-induced hyperalgesia in rats with incisional pain.Methods:Forty-eight male Sprague-Dawley rats with successful intrathecal and caudal vein catheterization, weighing 260-280 g, aged 2-3 months, were divided into 6 groups ( n= 8 each) using a random number table method: control group (group C), S1PR1 antagonist (FTY720) group (group F), remifentanil group (group R), remifentanil + S1PR1 antagonist (FTY720) group (group R+ F), remifentanil + incisional pain group (group R+ I), and remifentanil + incisional pain + S1PR1 antagonist (FTY720) group (group R+ I+ F). In C group, normal saline 0.1 μg·kg -1·min -1 was intravenously infused for 60 min. In R group, remifentanil 1.0 μg· kg -1·min -1 was infused for 60 min through the caudal vein. In F group, FTY720 3 nmol was intrathecally injected, and 10 min later normal saline 1.0 μg· kg -1·min -1 was infused for 60 min via the caudal vein. In R+ F group, FTY720 3 nmol was intrathecally injected, and 10 min later remifentanil 1.0 μg· kg -1·min -1 was infused for 60 min through the caudal vein. In R+ I group, remifentanil 1.0 μg·kg -1·min -1 was infused for 60 min through the caudal vein while the model of incisional pain was developed. In R+ I+ F group, FTY720 3 nmol was intrathecally injected, 10 min later the incisional pain model was prepared, and remifentanil 1.0 μg·kg -1·min -1 was injected for 60 min through the caudal vein at the same time. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 24 h before remifentanil or normal saline infusion (T 0) and 2, 6, 24 and 48 h after stopping remifentanil or normal saline infusion (T 1-4). Rats were sacrificed after the last measurement of pain threshold, and the L 4-6 segments of dorsal root ganglion were taken for determination of the expression of S1PR1, NOD-like receptor thermal protein domain-associated protein 3 (NLRP3), interleukin-1β (IL-1β) and glutamate transporter-1 (GLT-1) protein and mRNA (by Western blot and quantitative polymerase chain reaction). Results:Compared with C group, the MWT was significantly decreased and TWL was shortened at T 1-4, the expression of S1PR1, NLRP3 and IL-1β protein and mRNA in dorsal root ganglion was up-regulated, and the expression of GLT-1 protein and mRNA in dorsal root ganglion was down-regulated in R group ( P<0.05), and no significant change was found in the parameters mentioned above in group F ( P>0.05). Compared with R group, MWT was significantly decreased and TWL was shortened at T 1-4, the expression of S1PR1, NLRP3 and IL-1β protein and mRNA in dorsal root ganglion was up-regulated, and GLT-1 protein and mRNA expression in dorsal root ganglion was down-regulated in R+ I group, and MWT was significantly increased and TWL was prolonged at T 1-4, the expression of S1PR1, NLRP3 and IL-1β protein and mRNA in the dorsal root ganglion was down-regulated, and GLT-1 protein and mRNA expression in the dorsal root ganglion was up-regulated in R+ F group ( P<0.05). Compared with R+ I group, MWT was significantly increased and TWL was prolonged at T 1-4, the expression of S1PR1, NLRP3 and IL-1β protein and mRNA in the dorsal root ganglion was down-regulated, and the expression of GLT-1 protein and mRNA in the dorsal root ganglion was up-regulated in R+ I+ F group( P<0.05). Conclusions:The mechanism by which remifentanil induces hyperalgesia is associated with up-regulation of S1PR1 expression, activation of inflammatory factors, and down-regulation of GLT-1 expression in the rats with incisional pain.

Clinical Efficacy of Huanglian Jiedutang in Adjuvant Treatment of Acute Cerebral Infarction Complicated with Gastric Motility Disorder / 中国实验方剂学杂志

ObjectiveTo explore the clinical efficacy of Huanglian Jiedutang as an adjunctive treatment for acute cerebral infarction complicated with gastric motility disorder. MethodSixty patients with acute cerebral infarction complicated with gastric motility disorder with fire toxin syndrome were randomly divided into a western medicine control group （control group） and a traditional Chinese medicine （TCM） combined treatment group （observation group）， with 30 cases in each group. The control group received basic treatment for cerebral infarction and relevant western medical symptomatic treatment based on the patients' gastrointestinal symptoms. The observation group received Huanglian Jiedutang in addition to the treatment provided to the control group. The treatment course was 7 days. Neurological deficit scores and gastrointestinal dysfunction scores were assessed in both groups before treatment and on the 4th and 7th days of treatment. Gastrointestinal electrographic parameters， serum citrulline （CIT）， and motilin （MTL） levels were measured in both groups before treatment and on the 7th day of treatment. Clinical efficacy was compared between the two groups. ResultCompared with the baseline in both groups， the neurological deficit scores and gastrointestinal dysfunction scores were significantly reduced on the 4th and 7th days of treatment （P<0.05）. The reductions in these scores were more significant on the 7th day compared with those on the 4th day of treatment （P<0.05）. On the 4th and 7th days of treatment， the observation group showed a significantly greater reduction in neurological deficit scores and gastrointestinal dysfunction scores compared with the control group （P<0.05）. On the 7th day of treatment， compared with the baseline， both groups showed a significant increase in gastric antral and gastric body electric wave amplitudes as well as serum CIT and MTL levels （P<0.05）， and there were no statistically significant differences in the frequency of gastric antral and gastric body electric waves. On the 7th day of treatment， compared with the control group， the observation group had a significant increase in gastric antral and gastric body electric wave amplitudes as well as serum CIT and MTL levels （P<0.05）， and there were no statistically significant differences in the frequency of gastric antral and gastric body electric waves. After 7 days of treatment， the total effective rate in the observation group was 90.00% （27/30）， higher than 76.67% （23/30） in the control group， but the difference was not statistically significant. ConclusionAdjunctive treatment with Huanglian Jiedutang can effectively improve the symptoms of neurological function impairment and gastrointestinal dysfunction in patients with acute cerebral infarction complicated with gastric motility disorder， increase gastric antral and gastric body electric wave amplitudes， improve gastric motility disorder， and increase serum CIT and MTL levels， thereby improving the imbalanced secretion function of the gastrointestinal tract.

An ACE2-blocking antibody confers broad neutralization and protection against Omicron and other SARS-CoV-2 variants

The ongoing evolution of SARS-CoV-2 has resulted in the emergence of Omicron, which displays striking immune escape potential. Many of its mutations localize to the spike protein ACE2 receptor-binding domain, annulling the neutralizing activity of most therapeutic monoclonal antibodies. Here we describe a receptor-blocking human monoclonal antibody, 87G7, that retains ultrapotent neutralization against SARS-CoV-2 variants including the Alpha, Beta, Gamma, Delta and Omicron (BA.1/BA.2) Variants-of-Concern (VOCs). Structural analysis reveals that 87G7 targets a patch of hydrophobic residues in the ACE2-binding site that are highly conserved in SARS-CoV-2 variants, explaining its broad neutralization capacity. 87G7 protects mice and/or hamsters against challenge with all current SARS-CoV-2 VOCs. Our findings may aid the development of sustainable antibody-based strategies against COVID-19 that are more resilient to SARS-CoV-2 antigenic diversity. One sentence summaryA human monoclonal antibody confers broad neutralization and protection against Omicron and other SARS-CoV-2 variants

The level of regulatory T lymphocyte detected by flow cytometry in chimeric antigen receptor T cell immunotherapy and its clinical significance / 中华检验医学杂志

Objective:To investigate the characteristics of changes in peripheral blood regulatory T lymphocyte (Treg) levels in patients with B-cell lymphoma who received chimeric antigen receptor (CAR)-T cell immunotherapy, and the relationship between Treg levels and optimal efficacy and treatment response.Methods:The data of 23 patients with relapsed/refractory B-cell malignancies who received CD19/CD22 CAR-T cell immunotherapy in Wuhan Tongji Hospital from 2019 to 2021 were retrospectively studied. The enrolled patients were divided into complete remission (CR) group (8 cases), partial remission (PR) group (7 cases) and no response(NR) group (8 cases) according to Lugano′s revised lymphoma efficacy evaluation criteria. A total of 16 patients with B-cell lymphoma who did not receive CAR-T cell immunotherapy during the same period in Wuhan Tongji Hospital were collected as the control group.In different periods during CAR-T cell immunotherapy, multicolor flow cytometry(MFC) was used to dynamically detect peripheral blood the proportion of Treg in CD4 +T cells (Treg/CD4 +T), the proportion of lymphocytes (Treg/Lym), the proportion of Treg in white blood cells (Treg/WBC), and the absolute number of Treg (Treg#). The trend of Treg levels over time, as well as the differences in Treg levels in patients with different prognosis groups in different periods were analyzed.According to the proportion of Treg and the median level of absolute number within 1 to 15 days after CAR-T cell infusion, the patients were divided into a low-level group with 11 cases and a high-level group with 12 cases. The statistical differences in the peak value of CAR-T copy, iron protein, and IL-6 were compared between various groups. Independent samples t test, Mann-Whitney U test, Cox-Stuart trend existence test and one-way analysis of variance was used in statistical analysis. Results:In the 23 patients who received CAR-T cell immunotherapy, the mean values of Treg/CD4 +T and Treg/Lym before CAR-T cell infusion were (20.42±7.96)% and (13.61±7.13)%, respectively, which were significantly higher than that of the control group [(7.33±3.61)%, t=5.893, P<0.001; (1.91±0.90)%, t=6.53, P<0.001]. The number of Treg in the meantime was significantly lower [(1.81±1.52)/μl<(13.66±9.89)/μl, t=4.261, P<0.001]. After infusion, Treg/CD4 +T and Treg/Lym all remarkably decreased ( P<0.001),Treg/WBC increased significantly( P=0.01). The mean values of Treg/CD4 +T (12.87±1.93)%, Treg/Lym (6.35±2.84)%, and Treg/WBC (0.05±0.05)% in the patients with CR as the best response group were lower than those in the PR group [(29.68±5.49)%( P<0.01), (21.85±2.1)%( P<0.01), 0.50±0.69( P<0.05)] before CAR-T cell immunotherapy. Patients with lower mean Treg/CD4 +T within 1 to 15 days after reinfusion of CAR-T cells had higher peak CAR-T copy number ( P<0.05). Conclusion:Treg/CD4 +T and Treg/Lym were increased and then decreased during CAR-T treatment in B cell malignancies. The patients with lower proportions of Treg before infusion have favorable treatment efficacy. Besides, patients with lower Treg/CD4 +T after infusion have better CAR-T cell expansion. In the process of CAR-T cell immunotherapy, the use of MFC to dynamically monitor the proportion of Treg has certain clinical significance for the prediction of the optimal efficacy of immunotherapy and the prediction of treatment response.

Association of maternal adverse childhood experiences and parent child relationship in preschool children / 中国学校卫生

Objective@#To explore the relationship of maternal adverse childhood experiences(ACEs) with mother child relationship, so as to provide reference for parent child relationship and child health promotion.@*Methods@#In June 2021, children aged 3-6 years old of 36 kindergartens in three areas in Anhui Province were selected by stratified cluster sampling method, follow up data were collected in December 2021, and a total of 6 111 children were included in the study. Maternal ACEs and mother child relationship were respectively assessed using the Adverse Childhood Experiences International Questionnaire(ACEs-IQ)and the Child Parent Relationship Scale (CPRS). A multiple linear regression model was established to analyze the association of maternal ACEs and mother child relationship in preschool children.@*Results@#History of maternal childhood sexual abuse, physical neglect and peer bullying were negatively associated with mother child intimacy ( r =-0.03, -0.03, -0.03, P <0.05). Maternal emotional abuse, physical abuse, sexual abuse, emotional neglect, physical neglect, peer bullying, community violence, and total family dysfunction were positively associated with mother child dependence and mother child conflict ( r =0.09, 0.08, 0.05, 0.14, 0.06, 0.11, 0.08, 0.04; 0.18, 0.17, 0.07, 0.20, 0.11, 0.16, 0.12, 0.10, P <0.01). There was no statistically significant between all types of maternal ACEs and mother child intimacy in boys( P >0.05). Mothers with a history of physical abuse, sexual abuse and peer bullying had a statistically significant relationship between mother child intimacy in girls( β =-0.17, -0.62, -0.19, P <0.05). All types of maternal ACEs were positive predictors of mother child conflict between boys and girls( β =0.37-1.96, P <0.05). There was statistical significance between maternal childhood sexual abuse and mother child dependence of boys( β =0.53, P <0.05), but no statistical significance between maternal childhood sexual abuse and mother child dependence of girls( P >0.05). All other types of maternal ACEs were positive predictors of mother child dependence( β =0.09-0.41, P <0.05).@*Conclusion@#Maternal ACEs are associated with poor mother child relationship among preschool children, and maternal ACEs should be actively followed, which is of great significance for improving the parent child relationship and promoting child healthy development.

Association of maternal adverse childhood experiences and parenting styles with emotional behavior problems in preschool children / 中国学校卫生

Objective@#To explore the role of parenting style in the association of maternal adverse childhood experiences (ACEs) and emotional behavior problems (EBPs) in preschool children, so as to provide a reference for the prevention and control of EBPs in children.@*Methods@#A total of 6 111 children aged 3-6 years old from 36 kindergartens in 3 areas of Anhui Province in June 2021, follow up data were collected in December 2021. Maternal ACEs, mother child relationship and children EBPs were respectively assessed using the Adverse Childhood Experiences International Questionnaire(ACEs-IQ), Parental Rearing Style Scale and the difficulty score factor in the Chinese Strength and Difficulty Questionnaire(SDQ). The Bootstrap was used to examine the mediation effect of maternal parenting styles.@*Results@#Maternal ACEs were positively associated with child SDQ difficulty scores( r = 0.28, P <0.01). Negative parenting (indulgent, permissive, authoritarian and inconsistent) were positively correlated with maternal ACEs scores( r =0.28, 0.30, 0.21, 0.31) and child SDQ difficulty scores( r =0.25, 0.20, 0.20, 0.28)( P <0.01). Positive parenting (democracy) was negatively correlated with maternal ACEs and SDQ difficulty scores( r =-0.09, -0.29, P <0.01). After adjusting for confounding factors, the results of the mediation effect test of Bootstraping procedure showed that maternal parenting styles (indulgent, democracy, permissive, authoritarian and inconsistent) played a mediation role in maternal ACEs and EBPs of preschool children, and the mediation effects were respectively 19.13%, 7.34%, 24.88%, 12.05% and 26.83%.@*Conclusion@#Parenting styles play a mediating role in the association of maternal ACEs and EBPs in preschool children, and improving mothers negative parenting styles is of great significance to reduce EBPs in the offspring of maternal ACEs.

Relationship between maternal adverse childhood experiences and preschool children s emotional and behavioral problems / 中国学校卫生

Objective@#To explore the relationship and gender difference between maternal adverse childhood experiences (ACEs) with emotional and behavioral problems (EBPs) in preschool children, and to provide a reference to promote emotional and behavioral problems among preschool children.@*Methods@#Using the method of stratified cluster sampling, 9 647 children from kindergartens in three cities of Anhui Province were selected in June 2021. Maternal adverse childhood experiences were investigated with the WHO Adverse Childhood Experiences International Questionnaire(ACEs-IQ), preschool children s emotional and behavioral problems were investigated with Strengths and Difficulties Questionnaire.@*Results@#The detection rate of abnormal emotional behavior was 17.4%, emotional symptoms 16.3%, conduct problems 18.3%, hyperactivity 22.7%, peer problems 34.2%, prosocial behaviors 18.5%. Multivariate Logistic regression analysis showed that maternal adverse childhood experiences were positively associated with the risk of emotional and behavioral problems ( OR =1.51-2.97, P <0.01). Maternal cumulative adverse childhood experiences were also positively associated with the risk of emotional and behavioral problems( OR =3.13-9.61, P <0.01). The association of maternal emotional abuse, physical abuse and community violence with peer problems were stronger in boys than that of girls ( ROR =1.25, 1.26, 1.41, P <0.05).@*Conclusion@#The findings suggest maternal adverse childhood experiences were associated with emotional behavior problems among preschool children. Focusing on the maternal adverse childhood experiences is crucial for the prevention and control of childhood emotional and behavioral problems.

Relationship between family rearing style and 3-6 year old children s emotional and behavioral problems / 中国学校卫生

Objective@#To explore the relationship and gender difference between family rearing styles with emotional and behavior problems in preschool children aged 3-6 years, so as to provide reference for early prevention and intervention of children s emotional and behavior problems.@*Methods@#Using the method of stratified cluster sampling, 9 647 children aged 3-6 years old from 36 kindergartens in Wuhu, Lu an and Fuyang of Anhui Province were selected in June 2021. Primary caregivers were investigated with self designed questionnaire, Strengths and Difficulties Questionnaire (parents version) and Parental Rearing Style Scale.@*Results@#The detection rate of elevated SDQ total difficulty score was 6.5%, with boys (7.1%) higher than that of girls (5.8%). The detection rate of abnormal emotional behavior were significantly higher in children with high scores on doting, laissez faire, autocracy and inconsistency of family rearing style, compared with those in the low score group ( χ 2=210.32, 203.87, 102.70, 212.69, P <0.01 ), and the detection rate increased with the increase of score. However, the detection rate of abnormal emotional behavior in the high score group of democracy was significantly lower than that in the low score group ( χ 2=156.24, P <0.01), and the detection rate decreased with the increase of score. Logistic regression analysis showed that high level doting ( OR =4.31), laissez faire ( OR = 4.16), autocracy ( OR =3.36) and inconsistency ( OR =4.76) of family rearing style were associated with high risk of children s emotional behavior problems, while high level of democracy ( OR =0.34) in family rearing style was associated with low risk of emotional behavior problems. The comparison between boys and girls showed that the risk of emotional and behavioral problems in the indulgent rearing style of boys was significantly higher than that of girls ( OR =1.90, 2.13) ( P <0.05).@*Conclusion@#Family rearing styles are associated with emotional and behavioral problems among preschool children. Boys are sensitive to the negative impact of doting rearing style. Good rearing styles is beneficial to the prevention and control of children s emotional and behavioral problems.

Relationship between CCL21 and TREM2/DAP12 signaling pathways in spinal dorsal horn in remifentanil-induced hyperalgesia in mice with incisional pain / 中华麻醉学杂志

Objective:To evaluate the relationship between CCL21 and triggering receptor expressed on myeloid cells 2 (TREM2)/DNAX-activating protein of 12 kDa (DAP12) signaling pathways in the spinal dorsal horn in remifentanil-induced hyperalgesia in mice with incisional pain.Methods:Thirty-two SPF healthy male C57BL/6J mice, weighing 18-22 g, aged 8-10 weeks, were divided into 4 groups ( n=8 each) using a random number table method: control group (group C), CCL21 neutralizing antibody group (group anti-CCL21), remifentanil + incisional pain group (group R+ I), and CCL21 neutralizing antibody + remifentanil + incisional pain group (group anti-CCL21+ R+ I).A CCL21 neutralizing antibody 0.3 μg (diluted to 10 μl in normal saline) was intrathecally injected in anti-CCL21 and anti-CCL21+ R+ I groups twice a day.Normal saline 10 μl was intrathecally injected at the same time point twice a day in C and R+ I groups.Fifteen min after intrathecal injection, normal saline 0.1 ml was injected via the caudal vein for 4 consecutive times at an interval of 15 min in C and anti-CCL21 groups, and remifentanil 10 μg/kg (diluted to 0.1 ml in normal saline) was injected via the caudal vein for 4 consecutive times at an interval of 15 min in R+ I and anti-CCL21+ R+ I groups.The tail-flick latency (TFL) and mechanical paw withdrawal threshold (MWT) were measured at 24 h before remifentanil or normal saline injection (T 0) and 3, 6, 24 and 48 h after stopping injection of remifentanil or normal saline (T 1-4).The mice were sacrificed after the last measurement of pain threshold, and L 4-6 segments of the spinal cord were removed for determination of the expression of TREM2 and DAP12 protein and mRNA (by Western blot or quantitative real-time polymerase chain reaction). Results:Compared with group C, TFL was significantly shortened and MWT was decreased at T 1-4, and the expression of TREM2 and DAP12 protein and mRNA was up-regulated in group R+ I and R+ I+ anti-CCL21 ( P<0.05), and no significant change was found in the parameters mentioned above in group anti-CCL21 ( P>0.05).Compared with group R+ I, TFL was significantly prolonged and MWT was increased at T 1-4, and the expression of TREM2 and DAP12 protein and mRNA was down-regulated in group anti-CCL21+ R+ I ( P<0.05). Conclusions:CCL21 is involved in remifentanil-induced hyperalgesia by activating TREM2/DAP12 signaling pathways in the spinal dorsal horn of mice with incisional pain.

Draft Genome Assembly and Annotation for Cutaneotrichosporon dermatis NICC30027, an Oleaginous Yeast Capable of Simultaneous Glucose and Xylose Assimilation

The identification of oleaginous yeast species capable of simultaneously utilizing xylose and glucose as substrates to generate value-added biological products is an area of key economic interest. We have previously demonstrated that the Cutaneotrichosporon dermatisNICC30027 yeast strain is capable of simultaneously assimilating both xylose and glucose, resulting in considerable lipid accumulation. However, as no high-quality genome sequencing data or associated annotations for this strain are available at present, it remains challenging to study the metabolic mechanisms underlying this phenotype. Herein, we report a 39,305,439 bp draft genome assembly for C. dermatis NICC30027 comprised of 37 scaffolds, with 60.15% GC content. Within this genome, we identified 524 tRNAs, 142 sRNAs, 53 miRNAs, 28 snRNAs, and eight rRNA clusters. Moreover, repeat sequences totaling 1,032,129 bp in length were identified (2.63% of the genome), as were 14,238 unigenes that were 1,789.35 bp in length on average (64.82% of the genome). The NCBI non-redundant protein sequences (NR) database was employed to successfully annotate 11,795 of these unigenes, while 3,621 and 11,902 were annotated with the Swiss-Prot and TrEMBL databases, respectively. Unigenes were additionally subjected to pathway enrichment analyses using the Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Cluster of Orthologous Groups of proteins (COG), Clusters of orthologous groups for eukaryotic complete genomes (KOG), and Non-supervised Orthologous Groups (eggNOG) databases.Together, these results provide a foundation for future studies aimed at clarifying the mechanistic basis for the ability of C. dermatis NICC30027 to simultaneously utilize glucose and xylose to synthesize lipids.

Establishment of Quality Standard for Benchmark Samples of Liangditang / 中国实验方剂学杂志

ObjectiveTo establish the quality standard of Liangditang benchmark samples. MethodUltra performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-QTOF-MS） was used to qualitatively analyze the chemical composition of Liangditang on the basis of molecular and fragment ion peak information with cracking law. The mobile phase was methanol （A）-0.05% phosphate aqueous solution （B） for gradient elution （0-10 min， 5%-23.5%A； 10-20 min， 23.5%A； 20-58 min， 23.5%-63%A； 58-60 min， 63%-90%A）， the flow rate was 0.8 mL·min-1， and the detection wavelength was 254 nm. Electrospray ionization was employed under positive ion mode， the detection range was m/z 100-1 700. Key quality attributes and sources were determined by comparing with single medicine and reference substances. Through mass transfer analysis of multiple batches from decoction pieces to benchmark samples， high performance liquid chromatography （HPLC） for determining the contents of index components and HPLC detection of characteristic maps were established. Through the determination of 15 batches of benchmark samples， the content range of the index components and the common peaks of the characteristic map were determined. Thin layer chromatography （TLC） was applied to the identification of 5 medicines in the formula. Moisture and dry extract yield of the benchmark samples were determined by drying method. ResultA total of 27 compounds were inferred from the benchmark samples of Liangditang， among which 9 compounds were confirmed by comparison with the control， including catalpol， harpagide， gallic acid， albiflorin， paeoniflorin， verbascoside， angoroside C， cinnamic acid and harpagoside. A method for determining the characteristic maps of the benchmark samples were established and 13 peaks were assigned， and the characteristic peaks were mainly derived from wine-processed products of Rehmanniae Radix， Scrophulariae Radix and wine-processed products of Paeoniae Radix Alba. The similarity between the characteristic map of 15 batches of benchmark samples and the control characteristic map was >0.9. Methods for the determination of paeoniflorin， harpagoside， L-hydroxyproline and glycine were established， and the contents of these four components in 15 batches of benchmark samples were within ±30% of the corresponding mean value， and the transfer rate of decoction pieces to the benchmark samples was stable and controllable. TLC was established to identify 5 prescription drugs （except Ejiao） with two kinds of test solutions， and the results showed that the method had good specificity. The average dry extract yield was 48.06%， and the average moisture was 5.58%， which were within the range of ±10% and ±30% of their mean values， respectively. ConclusionThe quality standard of Liangditang benchmark samples was as follows：the similarity between the benchmark samples and the control characteristic map is >0.9， the contents of paeoniflorin， harpagoside， L-hydroxyproline and glycine are 217-403， 24-46， 634-1 178， 1 253-2 328 mg per dose， the dry extract yield is 43.0%-53.0%， the moisture is 4.0%-7.0%， under the set detection conditions， the benchmark samples have corresponding characteristic spots by comparing with the control herbs of 5 medicines. This quality standard is stable and reliable， which fills the gap in the quality control of Liangditang， and can provide a reference for the establishment of the quality standard of Liangditang granules.

The glycosylated extracellular domain of MUC1 protects against SARS-CoV-2 infection at the respiratory surface

Mucins play an essential role in protecting the respiratory tract against microbial infections. The heavily O-glycosylated gel-forming mucins MUC5AC and MUC5B eliminate pathogens by mucociliary clearance while transmembrane mucins MUC1, MUC4, and MUC16 restrict microbial invasion at the apical surface of the epithelium. In this study, we determined the impact of host mucins and mucin glycans on SARS-CoV-2 spike-mediated epithelial entry. Human lung epithelial Calu-3 cells have endogenous expression of the SARS-CoV-2 entry receptor ACE2 and express high levels of glycosylated MUC1 on the surface but not MUC4 and MUC16. Removal of the MUC1 extracellular domain (ED) using the O-glycan-specific mucinase StcE greatly enhanced spike binding and viral infection. By contrast, removal of mucin glycans sialic acid and fucose did not impact viral invasion. This study implicates the glycosylated ED of MUC1 as an important component of the host defense that restricts the severity of SARS-CoV-2 infection.

Multi-specific DARPin(R) therapeutics demonstrate very high potency against mutated SARS-CoV-2 variants in vitro

SARS-CoV-2 has infected millions of people globally and continues to undergo evolution. Emerging variants can be partially resistant to vaccine induced immunity and therapeutic antibodies, emphasizing the urgent need for accessible, broad-spectrum therapeutics. Here, we report a comprehensive study of ensovibep, the first trispecific clinical DARPin candidate, that can simultaneously engage all three units of the spike protein trimer to potently inhibit ACE2 interaction, as revealed by structural analyses. The cooperative binding of the individual modules enables ensovibep to retain inhibitory potency against all frequent SARS-CoV-2 variants, including Omicron BA.1 and BA.2, as of February 2022. Moreover, viral passaging experiments show that ensovibep, when used as a single agent, can prevent development of escape mutations comparably to a cocktail of monoclonal antibodies (mAb). Finally, we demonstrate that the very high in vitro antiviral potency also translates into significant therapeutic protection and reduction of pathogenesis in Roborovski dwarf hamsters infected with either the SARS-CoV-2 wild-type or the Alpha variant. In this model, ensovibep prevents fatality and provides substantial protection equivalent to the standard of care mAb cocktail. These results support further clinical evaluation and indicate that ensovibep could be a valuable alternative to mAb cocktails and other treatments for COVID-19.

Expanded circulating peripheral helper T cells in primary biliary cholangitis: Tph cells in PBC.

BACKGROUND: Peripheral helper T (TPH) cells, a recently defined subset of Th cells, promote B cell differentiation and antibody production in inflamed tissues. This study investigated whether circulating TPH cells are associated with primary biliary cholangitis (PBC), a typical organ-specific autoimmune disease. METHODS: Twenty PBC patients and 20 age- and sex-matched healthy controls (HCs) were recruited. The circulating TPH cell subsets were analyzed by flow cytometry, and the associations of TPH cells with disease activity and plasma cells were determined. Functional analysis was performed using a TPH and B cell coculture experiment. RESULTS: The frequencies of circulating TPH cells, ICOS+ TPH cells, and CD28+ TPH cells were increased in patients with PBC. Furthermore, the ICOS+ TPH cell level was higher in PBC patients with or without cirrhosis than in HCs, and the level decreased after treatment. Moreover, ICOS+ TPH cell levels correlated positively with specific clinical parameters (including anti-mitochondrial antibodies against M2 antigen (AMA-M2), IgM) and plasma cell levels, suggesting that the TPH cell activation status is associated with the severity of PBC. Coculture results revealed an enhanced ability of TPH cells from PBC patients to induce B cell differentiation. CONCLUSIONS: Elevated numbers of TPH cells may be involved in the pathogenesis of PBC, and the activation status of TPH cells is related to the severity of PBC. Additionally, TPH cells can be used as a useful biomarker for evaluating the progression of PBC and may serve as a therapeutic target for PBC patients in the future.

A Role for Transmembrane Protein 16C/Slack Impairment in Excitatory Nociceptive Synaptic Plasticity in the Pathogenesis of Remifentanil-induced Hyperalgesia in Rats / 神经科学通报·英文版

Remifentanil is widely used to control intraoperative pain. However, its analgesic effect is limited by the generation of postoperative hyperalgesia. In this study, we investigated whether the impairment of transmembrane protein 16C (TMEM16C)/Slack is required for α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic receptor (AMPAR) activation in remifentanil-induced postoperative hyperalgesia. Remifentanil anesthesia reduced the paw withdrawal threshold from 2 h to 48 h postoperatively, with a decrease in the expression of TMEM16C and Slack in the dorsal root ganglia (DRG) and spinal cord. Knockdown of TMEM16C in the DRG reduced the expression of Slack and elevated the basal peripheral sensitivity and AMPAR expression and function. Overexpression of TMEM16C in the DRG impaired remifentanil-induced ERK1/2 phosphorylation and behavioral hyperalgesia. AMPAR-mediated current and neuronal excitability were downregulated by TMEM16C overexpression in the spinal cord. Taken together, these findings suggest that TMEM16C/Slack regulation of excitatory synaptic plasticity via GluA1-containing AMPARs is critical in the pathogenesis of remifentanil-induced postoperative hyperalgesia in rats.

Difference of human epididymis protein 4 and carbohydrate antigen 125 in patients with ovarian cancer before and after menopause / 中国综合临床

Objective:To compare the difference of human epididymis protein 4(HE4) and carbohydrate antigen 125(CA125) indexes before and after menopause in ovarian cancer patients, and to evaluate the efficacy of risk of ovarian malignancy algorithm(ROMA) index in early diagnosis.Methods:The clinical data of 236 patients with ovarian cancer (malignant ovarian tumor group) who were hospitalized in Cancer Hospital of China Medical University from April 2018 to September 2019 were retrospectively analyzed, and 312 benign patients treated at the same period were selected as the control group (benign ovarian tumor group). On the basis of clinical pathological section confirmation, the serum samples were collected to detect HE4 and CA125, and the ROMA index was calculated according to the formula.The differences of HE4, CA125 and ROMA indexes before and after menopause, as well as the diagnostic efficacy of ROC survival curve of the three were compared, and the relationship between the different clinical characteristics of ovocarcinoma patients was evaluated to comprehensively analyze the early diagnostic effect of ROMA index.Results:In the serum of patients with malignant ovarian cancer, HE4, CA125 and ROMA index were abnormally high expression, and the difference between HE4 before(263.2(128.4, 385.6) pmol/L) and after menopause(354.8(179.5, 554.2) pmol/L) was statistically significant( Z=2.09, P=0.037). Through fitting the ROC curve: the AUC of premenopausal ROMA index was 0.939, and the AUC of postmenopausal ROMA index was 0.933, which were higher than HE4 (0.937, 0.906) and CA125 (0.913, 0.924), and their corresponding Youden correct diagnostic indexes were 0.800 and 0.742 respectively, which were higher than HE4 (0.788, 0.653) and CA125 (0.498, 0.565). The two categories of HE4 were associated with the occurrence of peritoneal effusion (the percentage of postmenopausal and premenopausal HE4<140 nmol/L and premenopausal HE4<70 nmol/L were 25.77%, 74.23% and 74.23% respectively), and the difference was statistically significant (χ 2=10.155, P=0.006). CA125 was associated with the classification of the largest diameter of parenchymal tumor and the occurrence of peritoneal effusion (all P<0.05). ROMA index was associated with menopause or not, classification of maximum diameter of mass, classification of maximum diameter of parenchymal tumor and occurrence of ascites, and the differences were statistically significant (all P<0.05). Conclusion:ROMA index has a good diagnostic efficiency in the diagnosis of malignant ovarian cancer, especially premenopausal patients with good diagnostic efficiency, for early diagnosis of ovarian cancer has a certain guiding value, the follow-up will further carry out confirmatory research.

Genetic characteristics of Brucella strains isolated from a human brucellosis epidemic in Baoding City of Hebei Province / 中华地方病学杂志

Objective:To analyze the genetic characteristics of Brucella strains isolated from a human brucellosis epidemic in Baoding City of Hebei Province, and to provide scientific basis for control and prevention of brucellosis. Methods:Brucella antibody was detected in a high-risk population ( n=22) of brucellosis in Baoding City in 2018 by using rose bengal plate test and tube agglutination test. Blood samples of patients with brucellosis were collected for Brucella isolation ( n=3). Conventional methods were used to identify the bacteria strains. Multiple locus variable-number tandem repeat analysis (MLVA) and multilocus sequence typing (MLST) were used to analyze the genetic characteristics of the strains. Results:The sero-prevalence of Brucella antibodies in high-risk populations was 4.55% (1/22). Three suspected Brucella strains were isolated from blood samples of three patients (No. BDY-1, BDY-2 and BDY-3) and were identified as Brucella melitensis biovar 3. The results of MLVA showed that strain BDY-1 and strain BDY-2 were the same genotype. Strain BDY-3 added two tandem repeats at the locus of Bruce04 and Bruce16, respectively, and lost three tandem repeats at the locus of Bruce30. The genotype of panel 1 (MLVA-8) and panel 1 + panel 2A (MLVA-11) were 42 and 116, respectively, which belonged to the "East Mediterranean group". They were closely clustered with Brucella melitensis biovar 3. MLST analysis showed that all isolated strains could be classified into a ST8 type. Conclusions:The isolated strains from the epidemic are Brucella melitensis biovar 3. In the future, the prevention and control of brucellosis in Baoding City should strengthen the health education and behavior intervention of the high-risk population.

A case of buttonhole puncture of anastomotic stump after occlusion of brachial artery-right atrium ACUSEAL arteriovenous graft / 中国实用护理杂志

Objective:To investigate the feasibility of using anastomotic stoma as a long-term vascular access to maintain hemodialysis in patients who cannot establish an effective vascular access after ACUSEAL graft occlusion.Methods:A rare case of brachial artery-right atrium ACUSEAL artificial blood vessel fistula bypass to establish vascular access occlusion, the use of artificial blood vessel anastomosis stoma to establish buttonhole puncture as a long-term vascular access and the corresponding nursing methods.Results:At present, the patients were treated with regular dialysis for 32 months and blunt needle puncture for 23 months.Conclusions:Brachial artery-right atrium ACUSEAL graft is a rare vascular access surgery. As a new type of access with depleted autologous vascular resources, the efficacy is not certain. Because it is different from the traditional vascular prosthesis surgery, once the thrombotic occlusion of the graft fails, the risk and difficulty of reopening by interventional or surgical methods are great. Therefore, it is a challenging and reasonable method to use the limited residual cavity of the anastomosis as a long-term vascular access after occlusion to prolong the service life of the graft.