Phosphorothioate-modified G-quadruplex as a signal-on dual-mode reporter for CRISPR/Cas12a-based portable detection of environmental pollutants.

BACKGROUND: Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a-powered biosensor with a G-quadruplex (G4) reporter offer the benefits of simplicity and sensitivity, making them extensively utilized in detection applications. However, these biosensors used for monitoring pollutants in environmental water samples may face the problem of high background signal and easy interference due to the "signal-off" output. It is obvious that a biosensor based on the CRISPR/Cas12a system and G4 with a "signal on" output mode needs to be designed for detecting environmental pollutants. RESULTS: By using phosphorothioate-modified G4 as a reporter and catalytic hairpin assembly (CHA) integrated with Cas12a as an amplification strategy, a "signal-on" colorimetric/photothermal biosensor (psG4-CHA/Cas) for portable detection of environmental pollutants was developed. With the help of functional nucleotides, the target pollutant (kanamycin or Pb2+) triggers a CHA reaction to produce numerous double-strand DNA, which can activate Cas12a's trans-cleavage activity. The active Cas12a cleaves locked DNA to release caged psG-rich sequences. Upon binding hemin, the psG-rich sequence forms a psG4/hemin complex, facilitating the oxidation of the colorless 3,3',5,5'-tetramethylbenzidine (TMB) into the blue photothermal agent (oxTMB). The smartphone was employed for portable colorimetric detection of kanamycin and Pb2+. The detection limits were found to be 100 pM for kanamycin and 50 pM for Pb2+. Detection of kanamycin and Pb2+ was also carried out using a portable thermometer with a detection limit of 10 pM for kanamycin and 8 pM for Pb2+. SIGNIFICANCE: Sensitive, selective, simple and robust detection of kanamycin and Pb2+ in environmental water samples is achieved with the psG4-CHA/Cas system. This system not only provides a new perspective on the development of efficient CRISPR/Cas12a-based "signal-on" designs, but also has a promising application for safeguarding human health and environmental monitoring.

Efficacy and Safety of Crisaborole Ointment 2% in Chinese Patients Aged ≥ 2 Years with Mild to Moderate Atopic Dermatitis.

INTRODUCTION: Atopic dermatitis (AD) is a chronic immuno-inflammatory skin disease. Crisaborole ointment, 2%, is a nonsteroidal phosphodiesterase 4 inhibitor approved for the treatment of mild to moderate AD. This post hoc analysis assesses the efficacy and safety of crisaborole in Chinese patients aged ≥ 2 years with mild to moderate AD. METHODS: We evaluated the efficacy and safety of crisaborole in Chinese patients from the vehicle-controlled, phase 3 CrisADe CLEAR study. Patients were randomly assigned 2:1 to receive crisaborole or vehicle twice daily, respectively, for 28 days. The primary endpoint was percent change from baseline in Eczema Area and Severity Index (EASI) total score at day 29. Key secondary endpoints were improvement in Investigator's Static Global Assessment (ISGA), ISGA success, and change from baseline in weekly average Peak Pruritus Numerical Rating Scale (PP-NRS) score. Adverse events were documented. RESULTS: Of 391 patients in the overall study, 237 were from China, 157 assigned to crisaborole and 80 assigned to vehicle. A greater reduction in percent change from baseline in EASI total score at day 29 was shown in the crisaborole vs. vehicle group (least squares mean [LSM]: -66.34 [95% (confidence interval) CI -71.55 to -61.12] vs. -50.18 [95% CI -58.02 to -42.34]). Response rates for achievement of ISGA improvement (43.2% [95% CI 35.4-51.1] vs. 33.4% [95% CI 22.5-44.2]) and ISGA success (31.7% [95% CI 24.3-39.0] vs. 21.5% [95% CI 12.1-30.9]) at day 29 were higher in the crisaborole vs. vehicle group. A greater reduction in change from baseline in weekly average PP-NRS score at week 4 was observed in the crisaborole vs. vehicle group (LSM: -1.98 [95% CI -2.34 to -1.62] vs. -1.08 [95% CI -1.63 to -0.53]). No new safety signals were observed. CONCLUSION: Crisaborole was effective and well tolerated in Chinese patients aged ≥ 2 years with mild to moderate AD. TRIAL REGISTRATION: ClinicalTrials.gov, NCT04360187.

Influence of overcooking on in vitro digestion and fermentation of ground beef and whole wheat bread.

Areas of char or overcooking commonly appear in foods people consume. It has been reported that overcooked food is harmful to human health. However, little research exists on the effect of overcooking on digestible protein and starch content and gut microbial fermentation. This study aimed to reveal the connection between overcooking and the content of digestible protein and starch, as well as its impact on gut microbial fermentation. Digestible protein in the standard cooked ground beef patty was significantly higher than the overcooked samples (p = 0.009). Standard-cooked whole wheat bread also showed a significantly higher digestible protein content compared with overcooked (p = 0.009). A significant difference was also found in digestible starch content between standard cooked and overcooked bread samples (p = 0.02). Overcooking decreased acetate, propionate, iso-butyrate, iso-valerate and ammonia production by the gut microbiota during fermentation of the beef sample, and decreased propionate and ammonia production during fermentation of the bread sample (p < 0.05). Interestingly, overcooking enhanced butyrate production by the microbiota during fermentation of the bread sample (24 h of fermentation, p < 0.001; 48 h of fermentation, p = 0.02), while no significant difference was found between overcooked and standard cooked beef samples (24 h of fermentation, p = 0.15; 48 h of fermentation, p = 0.4). Overcooking resulted in reductions in many Pseudomonadota and favored several Bacillota, especially Ruminococcaceae and Oscillospiraceae, which contain butyrate producers. Overall, overcooking reduced digestible protein, digestible starch, and fermentation of proteins. Unexpectedly, overcooking induced several purportedly favorable effects on the gut microbiota due to the decreased protein fermentation, which, in future studies, should be weighed against the previous reports that overcooking is deleterious to human health.

Analysis of the factors influencing the effectiveness of local government's purchase of older adults care services - a grounded theory study based on typical cases.

Background: Population aging is a basic national condition in China at present and for a long time to come, forcing the country to accelerate the pace of building its public older adults care system. The government's purchase of older adults care services has become an effective way to make up for the lack of the family's older adults care function, to which the Chinese government attaches particular importance. The article selects 11 typical cases from the excellent case base released by the Chinese Ministry of Civil Affairs officials in 2022 to study the influencing factors of the effect of local government purchase of older adults care service supply. Methods: NVivo data analysis tools have significant advantages in retrieving, analyzing and coding data more efficiently and accurately, which helps to construct theoretical propositions and formulate hypotheses to be tested in qualitative research. The study intends to adopt the grounded theory approach to analyze the text with the help of NVivo12 software, to condense the practice mechanism of local governments' purchasing of older adults care services and to construct a relational model. Results: Taking "the supply effect of local government purchasing older adults services" as the main logic line, the article summarizes the four main influencing factors of the supply effect of government purchasing older adults services: the real demand of the society, the government's power and responsibility system, the government's governance ability, and the society's acceptance ability. Conclusion: The sense of gain, happiness and security of the older adults group is the starting point and landing point of the older adults service policy formulation and implementation. Policy guidance and decision-making have an important impact on the quality of the supply of older adults care services and the development of the older adults care services industry. Clarifying the direction of policy guidance, reflecting the comprehensive efficiency of government governance and utilizing the professional advantages of social forces, is the key to improving the effectiveness of the government's purchase of older adults care services.

Characterization of the Molecular Events Underlying the Establishment of Axillary Meristem Region in Pepper.

Plant architecture is a major motif of plant diversity, and shoot branching patterns primarily determine the aerial architecture of plants. In this study, we identified an inbred pepper line with fewer lateral branches, 20C1734, which was free of lateral branches at the middle and upper nodes of the main stem with smooth and flat leaf axils. Successive leaf axil sections confirmed that in normal pepper plants, for either node n, Pn (Primordium n) < 1 cm and Pn+1 < 1 cm were the critical periods between the identification of axillary meristems and the establishment of the region, whereas Pn+3 < 1 cm was fully developed and formed a completely new organ. In 20C1734, the normal axillary meristematic tissue region establishment and meristematic cell identity confirmation could not be performed on the axils without axillary buds. Comparative transcriptome analysis revealed that "auxin-activated signaling pathway", "response to auxin", "response to abscisic acid", "auxin biosynthetic process", and the biosynthesis of the terms/pathways, such as "secondary metabolites", were differentially enriched in different types of leaf axils at critical periods of axillary meristem development. The accuracy of RNA-seq was verified using RT-PCR for some genes in the pathway. Several differentially expressed genes (DEGs) related to endogenous phytohormones were targeted, including several genes of the PINs family. The endogenous hormone assay showed extremely high levels of IAA and ABA in leaf axils without axillary buds. ABA content in particular was unusually high. At the same time, there is no regular change in IAA level in this type of leaf axils (normal leaf axils will be accompanied by AM formation and IAA content will be low). Based on this, we speculated that the contents of endogenous hormones IAA and ABA in 20C1734 plant increased sharply, which led to the abnormal expression of genes in related pathways, which affected the formation of Ams in leaf axils in the middle and late vegetative growth period, and finally, nodes without axillary buds and side branches appeared.

Efficacy and safety of crisaborole ointment in Chinese and Japanese patients aged ≥2 years with mild-to-moderate atopic dermatitis.

Atopic dermatitis is a chronic inflammatory skin disease with a significant impact on the overall wellbeing of patients and their families. Crisaborole ointment, 2%, is a nonsteroidal phosphodiesterase 4 inhibitor approved for the treatment of mild-to-moderate atopic dermatitis in multiple countries. However, in the key pivotal trials, a low proportion of the overall patient population was Asian, therefore the safety and efficacy of crisaborole in the Asian population with atopic dermatitis remains unclear. CrisADe CLEAR was a multicenter, randomized, double-blind, vehicle-controlled, phase 3 study (NCT04360187) to assess the efficacy and safety of crisaborole ointment in Chinese and Japanese patients aged ≥2 years with mild-to-moderate atopic dermatitis involving ≥5% treatable body surface area. Patients were randomly assigned 2:1 to receive crisaborole or vehicle twice daily for 28 days. The primary endpoint was percentage change from baseline in the Eczema Area and Severity Index total score at day 29. Additional endpoints were improvement and success per Investigator's Static Global Assessment score at day 29 and change from baseline on the Peak Pruritus Numerical Rating Scale at week 4. Safety was assessed using rates of treatment emergent adverse events, serious adverse events, and clinically significant changes in vital signs and clinical laboratory parameters. Crisaborole-treated patients showed a significantly greater reduction versus vehicle in percentage change from baseline in Eczema Area and Severity Index total score at day 29 (P = 0.0002). Response rates for achievement of Investigator's Static Global Assessment improvement and success at day 29 were significantly higher for patients treated with crisaborole versus vehicle (P = 0.0124 and P = 0.0078, respectively). Crisaborole-treated patients showed a significantly greater reduction versus vehicle in change from baseline on the Peak Pruritus Numerical Rating Scale at week 4 (P = 0.0009). No new safety signals were identified. Treatment with crisaborole was effective and well tolerated in Chinese and Japanese patients with mild-to-moderate atopic dermatitis.

IgG4-related kidney disease complicated with retroperitoneal fibrosis: A case report.

BACKGROUND: IgG4-related disease (IgG4-RD) is a chronic fibrotic disease mediated by immunity recognized by clinicians in recent years. When the kidney is involved, it is called IgG4-related kidney disease (IgG4-RKD). IgG4-related tubulointerstitial nephritis (IgG4-TIN) is a representative manifestation of IgG4-RKD. IgG4-TIN can cause obstructive nephropathy complicated by retroperitoneal fibrosis (RPF). Cases of IgG4-TIN complicated with RPF are rare. Glucocorticoids are the first-line therapeutic medication for IgG4-RD and can significantly improve renal function. CASE SUMMARY: Herein, we report the case of a 56-year-old man with IgG4-RKD complicated with RPF. The patient presented to the hospital with complaints of elevated serum creatinine (Cr), nausea, and vomiting. During hospitalization, Cr was 1448.6 µmol/L, and serum IgG4 was increased. A total abdominal computed tomography (CT) scan and enhanced CT scan obviously indicated RPF. Although this patient had a long course and renal insufficiency, we performed a kidney biopsy. Renal biopsy showed that the renal tubulointerstitium had focal plasma cell infiltration and increased lymphocyte infiltration accompanied by fibrosis. After combining the biopsy results with immunohistochemistry, it was found that the absolute number of positive IgG4+ cells per high power field exceeded 10, and the ratio of IgG4/IgG was over 40%. Finally, the patient was diagnosed with IgG4-TIN complicated with RPF and given glucocorticoids as long-term maintenance therapy, helping him keep out of dialysis. After a follow-up of 19 mo, the patient had recovered well. Previous literature on IgG4-RKD and RPF was retrieved from PubMed to characterize the clinical and pathological features and to identify the diagnosis and treatment of IgG4-RKD. CONCLUSION: Our case report demonstrates the clinical characteristics of IgG4-RKD complicated with RPF. Serum IgG4 is a favorable indicator for screening. Performing renal biopsy actively plays a vital role in diagnosis and treatment, even if the patient has a long course and manifests with renal insufficiency. It is remarkable to treat IgG4-RKD with glucocorticoids. Hence, early diagnosis and targeted therapy are essential for reversing renal function and improving extrarenal manifestations in patients with IgG4-RKD.

PAM-free cascaded strand displacement coupled with CRISPR-Cas12a for amplified electrochemical detection of SARS-CoV-2 RNA.

The early diagnosis of coronavirus disease 2019 (COVID-19) is dependent on the specific and sensitive detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA. Herein, we develop a highly sensitive and specific electrochemical biosensor for SARS-CoV-2 target RNA detection based on the integration of protospacer adjacent motif (PAM)-free cascaded toehold-mediated strand displacement reaction (TSDR) and CRISPR-Cas12a (PfTSDR-CRISPR). In this study, each target is transformed into multiple DNA substrates with bubble structure in the seed region by the cascaded TSDR, which can directly hybridize with guide RNA (gRNA) without PAM requirement and then activate CRISPR-Cas12a's trans-cleavage activity. Subsequently, the hairpin DNA modified with methylene blue (MB-HP) is cleaved by activated CRISPR-Cas12a. Therefore, as MB leaves the electrode surface, a decreased current signal is obtained. With the involvement of PAM-free cascaded TSDRs and CRISPR-Cas12a amplification strategy, the PfTSDR-CRISPR-based electrochemical biosensor achieves the detection of target RNA as low as 40 aM. The biosensor has high sequence specificity, reliability and robustness. Thanks to the PAM-free cascaded TSDR, the biosensor can achieve universal detection of different target RNA without redesigning gRNA sequence of CRISPR-Cas12a. In addition, this biosensor successfully detects SARS-CoV-2 target RNA in complex samples, which highlights its potential for diagnosing COVID-19.

Imaging of the Abdominal Wall Sinus Tract: Comparison of Computed Tomography Sinography and X-Ray Sinography.

OBJECTIVE: The study aims to clarify the comparative benefits of X-ray sinography and computed tomography (CT) sinography in assessing the abdominal wall sinus tract. METHODS: In this cross-sectional study, patients in our hospital with an abdominal wall sinus tract who had received both X-ray sinography and CT sinography from January 2018 to January 2021 were enrolled. The intraoperative findings were used as the gold standard to calculate the accuracy of the two methods. Kappa statistic was employed to evaluate the concordance between the two methods and the intraoperative findings. Differences in diameters measured on X-ray sinography and CT sinography images were analyzed using the Wilcoxon signed rank test. RESULTS: The study sample consisted of 74 patients. The accuracy of the CT sinography in diagnosing the extent of the sinus invasion was 85.1%, while the accuracy of the X-ray sinography was 59.5%. For the sinus confined to the abdominal wall (Kappa: 0.783 VS 0.248), the sinus extending into the abdominal cavity (Kappa: 0.734 VS 0.339), and the sinus with fistula formation (Kappa: 0788 VS 0.496), the consistency of the CT sinography and surgery were significantly better than that of the X-ray sinography. Diameters of the sinus tract measured on CT images were statistically larger than the diameters measured on X-ray sinography (P<0.001). CONCLUSION: CT sinography has significant advantages to X-ray sinography in depicting the extent of the abdominal wall sinus tract and the presence of a fistula.

Mapping of the AgWp1 gene for the white petiole in celery (Apium graveolens L.).

Celery (Apium graveolens L.) is one of the most popular leafy vegetables worldwide. The main edible parts of celery are the leaf blade and especially the petiole, which typically has a white, green and red color. To date, there are very few reports about the inheritance and gene cloning of celery petiole color. In this study, bulked segregant analysis-sequencing (BSA-Seq) and fine mapping were conducted to delimit the white petiole (wp1) loci into a 668.5-kb region on Chr04. In this region, AgWp1 is a homolog of a DAG protein in Antirrhinum majus and a MORF9 protein in Arabidopsis, and both proteins are involved in chloroplast development. Sequencing alignment shows that there is a 27-bp insertion in the 3'-utr region in AgWp1 in the white petiole. Gene expression analysis indicated that the expression level of AgWp1 in the green petiole was much higher than that in the white petiole. Further cosegregation revealed that the 27-bp insertion was completely cosegregated with the petiole color in 45 observed celery varieties. Therefore, AgWp1 was considered to be the candidate gene controlling the white petiole in celery. Our results could not only improve the efficiency and accuracy of celery breeding but also help in understanding the mechanism of chlorophyll synthesis and chloroplast development in celery.

Ultrasonography Combined with Blood Biochemistry on the Early Diagnosis of Diabetic Kidney Disease.

Objective: Diabetic kidney disease (DKD) has been well recognized as a microvascular complication of diabetes mellitus. Perfusion of intrarenal arteries is closely related with development of DKD. The aim of the present study was to investigate relation of ultrasonography performance of intrarenal arteries and grade of DKD. Methods: From May to December at 2021, a total of 54 DKD patients and 36 non-DKD cases were recruited. Ultrasonography performance of intrarenal and arteries at lower extremity was examined by high-resolution ultrasound diagnostic equipment; maximum (Vsmax) as well as minimum (Vdmin) blood velocity of arteries were recorded, and resistance index (RI) of arteries were calculated. Blood routine and biochemical parameters were determined from clinical laboratory of our hospital. Results: According to eGFR grading, 42.50% of the 54 DKD cases are at Grade 1, and 18.52%, 11.11%, 9.26%, and 18.52% cases were at Grade 2, 3a, 3b, and 4-5, respectively. Blood urea and creatinine were significantly positively related with progress of DKD, while level of Hb was negatively related with DKD. By ultrasonography; we found that Vsmax and Vdmin of main renal artery (MRA), segmental renal artery (SRA), and interlobular renal artery (IRA) were significantly reduced compared with healthy cases; IR of the above arteries was dramatically elevated, and changes of the above data were more obvious than that of lower extremity. Vdmin of MRA, SRA, and IRA was negatively related with grading of DKD, while RI was positively related with the grading. Converging from RI and level of Hb, we found that the level of Hb is positively related with healthy status of the kidney, while RI of the arteries is negatively with that. Conclusions: Resistance index (RI) of intrarenal arteries, obtained from ultrasonography combining with level of hemoglobin (Hb), is the predictor of progress of DKD.

End-Use Quality of Historical and Modern Winter Wheats Adapted to the Great Plains of the United States.

Improving milling and baking properties is important during wheat breeding. To determine changes in milling and baking quality of hard winter wheat, 23 adapted cultivars released in the Great Plains between 1870 and 2013 were grown in triplicate in a single location (Mead, NE, USA) over two crop years (2018 and 2019). Grain yield and kernel hardness index increased by release year (p < 0.05). The observed increase in hardness index was accompanied by a decrease in percent soft kernels (p < 0.05). Diameter and weight decreased with release year in 2019 (p < 0.05), and their standard deviation increased with the release year (p < 0.05). Flour protein content decreased with release year (p < 0.05) and dough mixing quality increased (p < 0.05). No significant relationship was found for baking property variables, but bran water retention capacity (BWRC), which is correlated with whole wheat bread quality, increased with release year (p < 0.05). In conclusion, wheat kernels have become harder but more variable in shape over a century of breeding. Mixing quality showed significant improvements, and loaf volume and firmness remained constant, even in the presence of a decrease in protein concentration. Bran quality decreased across release year, which may have implications for whole grain baking quality and milling productivity.

Chromoblastomycosis: A case series from Eastern China.

Chromoblastomycosis (CBM) is a chronic fungal infection of the cutaneous and subcutaneous tissues caused by brown pigmented fungi. Fonsecaea monophora is one of the most common pathogens of CBM in China. Most formal cases have been reported from Southern China, however, the infection is not uncommon in Eastern China where very few case series are available. To describe the clinical aspects of CBM, we report a series of 11 cases between 2018 and 2021 at a single medical center in Eastern China. The patients were predominately male (n = 9) and the disease duration ranged from 3 months to 20 years. Plaque type lesions were the most common clinical manifestations. There were 7 cases of mild forms and 3 cases of severe forms. Among the 3 severe cases, one case gave up treatment due to economic poverty; one case did not respond to a 1-year systemic treatmen; one case was cured by combination therapy of 10 months. Other cases were cured by treatment with antifungal agents. All cases of direct mycological examination were positive. All isolates were identified by morphology and sequencing of the the ITS regions of ribosomal DNA, Ten were F. monophora and 1 was Cladophialophora carrionii. All cases had been evaluated at other clinics, where 8 cases were misdiagnosed as other diseases. As a neglected tropical disease (NTD), CBM is still a major challenge in the field of dermatology, especially in its severe clinical forms. As an effective and simple diagnostic method of CBM, direct microscopic examination should be further promoted in rural hospitals.

Determining the optimal stage for cryopreservation of human embryonic stem cell-derived retinal pigment epithelial cells.

BACKGROUND: Human embryonic stem cell-derived retinal pigment epithelial cells (hESC-derived RPE) are a promising source for cell-replacement therapy to treat retinal degenerative diseases, but research on RPE cryopreservation is limited. This study aimed to determine the best phase for RPE cryopreservation to preserve the post-thaw function and uncover the mechanism underlying RPE freezing tolerance. METHODS: hESC-derived RPE cells were cryopreserved at various time points after seeding. After thawing, the survival and attachment rates, RPE marker gene expression, apical-basal polarity, PEDF secretion, transepithelial resistance, and phagocytotic ability of post-thaw RPE cells were evaluated. RNA sequencing was performed on RPE cells at three-time points, differentially expressed genes were identified, and gene ontology, Kyoto encyclopedia of genes and genomes, and protein-protein interaction analyses were used to investigate the key pathways or molecules associated with RPE cell freezing tolerance. RESULTS: RPE frozen at passage 2 day 5 (P2D5) had the highest cell viability and attachment after thawing. They also retained properly localized expression of RPE marker genes and biological functions such as PEDF secretion, high transepithelial resistance, and phagocytic ability. The RNA-sequencing analysis revealed that RPE cells at P2D5 expressed high levels of cell cycle/DNA replication and ECM binding associated genes, as well as THBS1, which may serve as a possible hub gene involved in freezing tolerance. We also confirmed that the RPE cells at P2D5 were in the exponential stage with active DNA replication. CONCLUSIONS: We propose that freezing hESC-derived RPE cells during their exponential phase results in the best post-thawing outcome in terms of cell viability and preservation of RPE cell properties and functions. The high expression levels of the cell cycle and ECM binding associated genes, particularly THBS1, may contribute to better cell recovery at this stage.

Carbohydrate utilization by the gut microbiome determines host health responsiveness to whole grain type and processing methods.

Little is known about how interactions among grain processing, grain type, and carbohydrate utilization (CU) by the microbiome influence the health benefits of whole grains. Therefore, two whole grains - brown rice and whole wheat - and two processing methods - boiling (porridge) and extrusion - were studied for their effects on host metabolic outcomes in mice harboring human microbiomes previously shown in vitro to have high or low CU. Mice carrying either microbiome experienced increases in body weight and glycemia when consuming Western diets supplemented with extruded grains versus porridge. However, mice with the high but not low CU microbiome also gained more weight and fat over time and were less glucose tolerant when consuming extruded grain diets. In high CU microbiome mice, the exacerbated negative health outcomes associated with extrusion were related to altered abundances of Lachnospiraceae and Ruminococcaceae as well as elevated sugar degradation and colonic acetate production. The amplicon sequence variants (ASVs) associated with extruded and porridge diets in this in vivo study were not the same as those identified in our prior in vitro study; however, the predicted functions were highly correlated. In conclusion, mice harboring both high and low CU microbiomes responded to the whole grain diets similarly, except the high CU microbiome mice exhibited exacerbated effects due to excessive acetate production, indicating that CU by the microbiome is linked to host metabolic health outcomes. Our work demonstrates that a greater understanding of food processing effects on the microbiome is necessary for developing foods that promote rather than diminish host health.Abbreviations: CU- carbohydrate utilization; SCFA- short-chain fatty acids; GF- germ-free; HMA, human-microbiome associated; ipGTT- intraperitoneal glucose tolerance test; HOMA-IR- Homeostatic Model Assessment for Insulin Resistance; AUC- area under the glycemia curve; ASV- amplicon sequence variant; lf- low-fat; wd- Western diet; wd_wwp- Western diet containing whole wheat porridge; wd_wwe- Western diet containing whole wheat extrudate; wd_bre- Western diet containing brown rice extrudate; wd_extr- Western diet containing either whole wheat or brown rice extrudate.

Loci underlying leaf agronomic traits identified by re-sequencing celery accessions based on an assembled genome.

Celery is one of the most popular vegetables in the world. The main edible parts of celery are the leaf blade and petiole. The celery petiole is usually green, red, or white, with a hollow or solid pith. However, the loci/genes controlling these petiole-related traits have not been reported. In this study, we present a chromosome-level celery genome assembly with a total size of 3.339 Gb. Simultaneous bursts of long-terminal repeats (78.43%) contributed greatly to the large genome size. Re-sequencing and population structure analysis of 79 celery accessions revealed that they could be divided into Chinese celery and Western celery. By combining genome-wide association studies (GWAS) and mapping data, we located the hollow petiole (hp) loci in an 807.6-kb region on chromosome 11. This study provides valuable resources for genetic research on celery and is also helpful for the identification and cloning of genes controlling leaf agronomic traits in celery.

Value of CT sinography and analysis of missed diagnosis and misdiagnosis for abdominal wall sinus.

BACKGROUND: The value of CT (computed tomography) sinography in evaluating abdominal wall sinus tracts is currently unclear. The present study aims to investigate the accuracy of CT sinography in diagnosing the extent of abdominal sinus and analyze the reasons for misdiagnosis. MATERIALS AND METHODS: 64 patients with abdominal sinus tract formation (including fistula) undergoing CT sinography in our hospital from January 2018 to November 2020 were retrospectively analyzed. The CT images were blindly and independently re-assessed by two radiologists with 5- and 18-years work experience, respectively. Whether the sinus tract was confined to the abdominal wall or had invaded the abdominal cavity, and whether there was fistula formation were evaluated. The accuracy of CT sinography in diagnosing sinus invasion in the abdominal cavity and fistula formation was calculated. The agreements of CT sinography-surgical results and inter-observer were assessed using weighted-kappa statistics. RESULTS: The weighted- Kappa of inter-observer agreement (0.825, P < 0.001) and CT sinography-surgical results (0.828, P < 0.001) were both perfect. The diagnostic accuracy, sensibility, and specificity of sinus tract confined to the abdominal wall were 90.6% (95% CI: 80.7-96.5), 85.7% (95% CI: 67.3-96.0), and 94.4% (95% CI: 81.3-99.3), respectively. The diagnostic accuracy, sensibility, and specificity of fistula formation were 93.8% (95% CI: 84.8-98.3), 89.5% (95% CI: 66.9-98.7), and 95.6% (95% CI: 84.9-99.5), respectively. A total of 4 cases of sinus tract confined to the abdominal wall were misdiagnosed as invading the abdominal cavity, 2 cases of sinus tract invading the abdominal cavity were misdiagnosed as confined to the abdominal wall, 2 cases of enterocutaneous fistula were missed, 1 case of enterocutaneous fistula was misdiagnosed, 1 case of vesico-cutaneous fistula was misdiagnosed, and no cases of vesico-cutaneous fistula were missed. CONCLUSIONS: CT sinography can accurately assess the extent of an abdominal sinus tract and reveal fistula formation, despite some inevitable misdiagnosis and missed diagnosis. Radiologists should find more clues to improve the diagnostic accuracy.

Curcumol Attenuates Endometriosis by Inhibiting the JAK2/STAT3 Signaling Pathway.

BACKGROUND Curcumol is a hydrogenated austenitic compound with hemiketal. In this study we evaluated the effects of curcumol on local inflammatory response, cell proliferation, and metastasis in endometriosis, and elucidated the underlying mechanisms. MATERIAL AND METHODS Ectopic endometrial stromal cells were treated with increasing doses of curcumol. The MTT assay was used to assess cell viability. FITC-labeled annexin-V/PI double-staining method and flow cytometry were used to determine cell apoptosis. Cell migration was evaluated using a wound healing assay. ELISA kits were used to detect the levels of TNF-alpha, IL-6, and IL-1ß. Western blot assay was used to examine the phosphorylation degree of JAK2 and STAT3 and the expression of Bax, Bcl2, and caspase-3 proteins. Autologous endometrial transplantation was used to establish a rat model to assess the anti-EMS effect of curcumol in vivo. RESULTS Curcumol can inhibit the proliferation of ectopic endometrial stromal cells, promote cell apoptosis, and weaken cell migration ability. Curcumol can reduce the expression of Bax and caspase-3 protein and increase the expression of Bcl2 protein. Curcumol also can inhibit the secretion of inflammatory cytokines, including tumor necrosis cytokines (TNF)-alpha, interleukin (IL)-6, and IL-1ß, by ectopic endometrial stromal cells. In addition, curcumol can also inhibit the phosphorylation of JAK2 and STAT3. In vivo experiments also proved that curcumol could inhibit the growth of ectopic lesions in EMS model rats. CONCLUSIONS Curcumol can inhibit the JAK2/STAT3 pathway, reduce the inflammatory cytokines secreted by ectopic endometrial stromal cells, inhibit cell proliferation and migration, and reduce the volume of ectopic lesions.

Identification of the Regulatory Genes of UV-B-Induced Anthocyanin Biosynthesis in Pepper Fruit.

Fruit peels of certain pepper (Capsicum annum L.) varieties accumulate a large amount of anthocyanins and exhibit purple color under medium-wave ultraviolet (UV-B) conditions, which severely impacts the commodity value of peppers. However, the regulatory mechanism of the above process has not been well studied so far. To explore which key genes are involved in this regulatory mechanism, pepper variety 19Q6100, the fruit peels of which turn purple under UV-B conditions, was investigated in this study. Transcription factors with expression levels significantly impacted by UV-B were identified by RNA-seq. Those genes may be involved in the regulation of UV-B-induced anthocyanin biosynthesis. Yeast one-hybrid results revealed that seven transcription factors, CabHLH143, CaMYB113, CabHLH137, CaMYBG, CaWRKY41, CaWRKY44 and CaWRKY53 directly bound to the putative promotor regions of the structural genes in the anthocyanin biosynthesis pathway. CaMYB113 was found to interact with CabHLH143 and CaHY5 by yeast two-hybrid assay, and those three genes may participate collaboratively in UV-B-induced anthocyanin biosynthesis in pepper fruit. Virus-induced gene silencing (VIGS) indicated that fruit peels of CaMYB113-silenced plants were unable to turn purple under UV-B conditions. These findings could deepen our understanding of UV-B-induced anthocyanin biosynthesis in pepper.

A Clinical Study of Minilaparoscopy in the Treatment of Cryptorchidism with an Ipsilateral Inguinal Hernia.

Introduction: The aim of this study was to investigate the safety and efficacy of performing minilaparoscopy compared with standard laparoscopy in the treatment of cryptorchidism with an ipsilateral inguinal hernia. Materials and Methods: In total, 46 patients with cryptorchidism and an ipsilateral inguinal hernia were admitted to the Urology and Hernia and Abdominal Wall Surgery Departments of Beijing Chaoyang Hospital between October 2009 and July 2019. They were assigned to two groups: Group M and Group S. In Group M, 24 patients underwent herniorrhaphy and orchiopexy using minilaparoscopy, and in Group S, 22 patients underwent herniorrhaphy and orchiopexy using standard laparoscopy. Surgeons chose the procedure at random, and the patients were blinded to the selected procedure. Results: Postoperative painkiller demand (P = .043) and first postoperative day Numerical Rating Scale scores (P = .032) were lower in Group M than Group S, and the average hospital stay was shorter (P = .041) in Group M. Furthermore, 21 of the 24 procedures in Group M were successful, 3 procedures of Group M were converted from mini- to standard laparoscopy, and all 22 procedures in Group S were successful. The Observer Scar Assessment Scale questionnaire results of Group M were significantly higher than for patients in Group S (P = .038). Conclusion: Our findings suggest that treatment of cryptorchidism with ipsilateral inguinal hernia using minilaparoscopy is as safe and effective as standard laparoscopy.