A 1-kb human CDCA8 promoter directs the spermatogonia-specific luciferase expression in adult testis.

Cell division cycle associated 8 (CDCA8) is a component of the chromosomal passenger complex and plays an essential role in mitosis, meiosis, cancer growth, and undifferentiated state of embryonic stem cells. However, its expression and role in adult tissues remain largely uncharacterized. Here, we studied the CDCA8 transcription in adult tissues by generating a transgenic mouse model, in which the luciferase was driven by a 1-kb human CDCA8 promoter. Our previous study showed that this 1-kb promoter was active enough to dictate reporter expression faithfully reflecting endogenous CDCA8 expression. Two founder mice carrying the transgene were identified. In vivo imaging and luciferase assays in tissue lysates revealed that CDCA8 promoter was highly activated and drove robust luciferase expression in testes. Subsequently, immunohistochemical and immunofluorescent staining showed that in adult transgenic testes, the expression of luciferase was restricted to a subset of spermatogonia that were located along the basement membrane and positive for the expression of GFRA1, a consensus marker for early undifferentiated spermatogonia. These findings for the first time indicate that the CDCA8 was transcriptionally activated in testis and thus may play a role in adult spermatogenesis. Moreover, the 1-kb CDCA8 promoter could be used for spermatogonia-specific gene expression in vivo and the transgenic lines constructed here could also be used for recovery of spermatogonia from adult testes.

Effects of surgery on survival of elderly patients with gallbladder cancer: A propensity score matching analysis of the SEER database.

Background: Surgery is the sole curative therapy for gallbladder cancer (GBC) patients. Confronting an aging society, the demand to treat elderly patients with GBC is increasing. But there are few reports on survival benefit in elderly GBC patients treated with surgery. Therefore, we designed this population-based study to assess the survival benefit of surgery in GBC patients aged 70 years or older. Methods: GBC patients aged 70 years or older were identified in the surveillance, epidemiology, and end results cancer (SEER) database from 2010 to 2017. A 1:1 propensity score matching (PSM) analysis was conducted to balance the baseline data of patients. Overall survival (OS) and cancer-specific survival (CSS) of patients were evaluated by Kaplan-Meier analysis and compared with log-rank test. Independent risk factors associated with OS and CSS were determined by univariate and multivariate Cox proportional hazard regression analyses and subgroup analysis were performed. Results: A total of 2055 GBC patients aged 70 years or older were included in our study, with 1734 patients underwent surgery. Before PSM, the age, AJCC stage, TNM stage, and chemotherapy were significantly different between the surgery and no-surgery group (all P<0.05). Patients with surgery had significantly longer OS and CSS than those without surgery (P<0.0001). After 1:1 PSM, the differences in clinicopathological characteristics were reduced (all P>0.05). Kaplan-Meier analysis also showed patients received surgery had significantly better OS and CSS (P<0.0001). Subgroup analysis further indicated that almost all subgroups received surgery had OS and CSS advantage, especially patients aged 70-84 years old. Finally, univariate and multivariate COX regression analyses showed that age, AJCC stage and T stage were independent prognostic factors for OS and CSS in patients undergoing surgery. Conclusion: Our study found that surgery significantly improved OS and CSS in GBC patients aged 70-84 years, but more prospective studies are needed to prove our findings.

Multi-omics analysis reveals focal adhesion characteristic associated tumor immune microenvironment in colon adenocarcinoma.

Colon adenocarcinoma (COAD) is one of the most frequent malignant lesions of the digestive system in humans, with an insidious onset. At the time of diagnosis, most of them have developed to the middle and late stages, and cancer cells have metastasized, and the prognosis is poor. Treatment options for progressive COAD are limited, and despite the promise of immunotherapy, immunotherapy response rates are low. The assembly and disaggregation of focal adhesion are critical for the directional migration of tumor cells to different sites, and it is unclear whether focal adhesion-related genes are involved in the development and prognosis of colon adenocarcinoma. This study aimed to investigate the role of focal adhesion genes in the occurrence and prognosis of COAD. We obtained datasets of COAD patients, including RNA-sequencing data and clinical information, from the TCGA and GEO databases (GSE17538 and GSE39582). Through CNMF clustering, two molecular subtypes with different expression patterns of focal adhesion genes were identified, and it was found that the molecular subtype with low expression of focal adhesion genes had better prognosis. Then the prediction signature was constructed by LASSO-Cox regression model, and the receiver operating characteristic (ROC) curve showed that the 4-gene signature had a good prediction effect on COAD 1-, 2-, and 3-year OS. Gene function enrichment analysis showed that the high-risk group was mainly enriched in immune and adhesion-related signaling pathways, suggesting that focal adhesion genes may affect the development and prognosis of COAD by regulating the immune microenvironment and tumor metastasis. The interaction between focal adhesion genes and immunity during the occurrence of COAD may help improve the response rate of immunotherapy, which also provides new ideas for the molecular mechanism and targeted therapy in COAD.

Multiple cholesterol granulomas of the breast: A case report and review of the literature.

RATIONALE: Cholesterol granuloma of the breast is a rare disease defined as chronic reactive inflammation of cholesterol crystals and foreign body giant cells. This disease can mimic breast cancer in the clinic as painless palpable hard nodules, and imaging shows irregular hypoechoic nodules with unclear boundaries. Therefore, the uncommon lesions can be easily misdiagnosed as breast cancer. Meanwhile, it can be easily neglected by clinicians because of poor understanding. PATIENT CONCERNS: In this report, we present a rare case of multiple cholesterol granulomas of the breast, which was analyzed retrospectively and combined with all 11 relevant available studies in the last 50 years. INTERVENTIONS: The patient had undergone multiple breast imaging inspections for breast nodules and had the local resection of nodules. DIAGNOSES: The patient was confirmed to have a final diagnosis of benign cholesterol granulomas but was initially considered as breast cancer. OUTCOMES: The patient did not complain of discomfort after surgery, and ultrasound reexamination 5 months after surgery showed no recurrence. LESSONS: By retrospective analysis, dynamic contrast-enhanced magnetic resonance imaging and core needle biopsy can synergistically help clinicians distinguish it from other breast disease. To raise awareness of such a rare disease and reduce related misdiagnoses, we summarize the characteristics of cholesterol granulomas and recommend appropriate novel diagnosis and treatment regimens for patients with cholesterol granulomas.

The effect of food vehicles on in vitro performance of pantoprazole sodium delayed release sprinkle formulation.

Certain patient populations, including children, the elderly or people with dysphagia, find swallowing whole medications such as tablets and capsules difficult. To facilitate oral administration of drugs in such patients, a common practice is to sprinkle the drug products (e.g., usually after crushing the tablet or opening the capsule) on food vehicles before consumption which improves swallowability. Thus, evaluation of the impact of food vehicles on the potency and stability of the administered drug product is important. The aim of the current study was to evaluate the physicochemical properties (viscosity, pH, and water content) of common food vehicles used for sprinkle administration (e.g., apple juice, applesauce, pudding, yogurt, and milk) and their impacts on the in vitro performance (i.e., dissolution) of pantoprazole sodium delayed release (DR) drug products. The food vehicles evaluated exhibited marked difference in viscosity, pH and water content. Notably, the pH of the food as well as the interaction between food vehicle pH and drug-food contact time were the most significant factors affecting the in vitro performance of pantoprazole sodium DR granules. For example, the dissolution of pantoprazole sodium DR granules sprinkled on food vehicles of low pH (e.g., apple juice or applesauce) for short durations remained unchanged compared with the control group (i.e., without mixing with food vehicles). However, use of high pH food vehicles (e.g., milk) with prolonged contact time (e.g., 2 h) resulted in accelerated pantoprazole release, drug degradation and loss of potency. Overall, a thorough assessment of physicochemical properties of food vehicles and formulation characteristics are a necessary part of the development of sprinkle formulations.

Reverse engineering the Ozurdex dexamethasone intravitreal implant.

Ozurdex is a biodegradable implant formulated for sustained-release delivery of the corticosteroid dexamethasone to the posterior segment of the eye. The small, rod-shaped implant is administered directly to the vitreous using a dedicated applicator, and releases drug for up to 6 months after administration. Sustained release is achieved by embedding dexamethasone in a matrix of 50:50 poly(lactic-co-glycolic acid) (PLGA). In this work, the Ozurdex implant was thoroughly characterized to enable the reverse engineering of a compositionally and structurally equivalent implant. Advanced imaging techniques such as scanning electron microscopy (SEM) and microcomputed tomography (microCT) revealed that the Ozurdex implant exhibits an irregular surface and an internal porosity of 6% due to a large number of discrete voids approximately 3 µm in diameter. Thermal and spectroscopic analyses showed limited interaction between the drug and the polymer, resulting in a two-phase system of dexamethasone crystals embedded within a PLGA matrix. Reverse-engineered implants with properties similar to Ozurdex were prepared using a two-step hot-melt extrusion process. The reverse-engineered implants exhibited a triphasic drug release profile similar to Ozurdex. This work seeks to provide insight into the manufacturing process and characterization of PLGA-based solid implants to support future generic product development.

Product Quality Research for Developing and Assessing Regulatory Submissions for Generic Cyclosporine Ophthalmic Emulsions.

Approval of the first generic 0.05% cyclosporine ophthalmic emulsion (COE) in the U.S. represents a milestone achievement of the science and research program in the U.S. Food and Drug Administration's Center for Drug Evaluation and Research (CDER). COE is a locally acting complex drug product indicated to increase tear production in patients whose production is presumed to be suppressed due to ocular inflammation associated with keratoconjunctivitis sicca. The path to approval required overcoming numerous scientific challenges to determining therapeutic equivalence to the reference listed drug. Researchers in CDER's Office of Pharmaceutical Quality and Office of Generic Drugs developed a quality by design approach to understand the effects of process and formulation variables on the product's critical quality attributes, including globule size distribution (GSD), turbidity, viscosity, zeta potential, surface tension, and osmolality. CDER researchers explored multiple techniques to perform physicochemical characterization and analyze the GSD including laser diffraction, nanoparticle tracking analysis, cryogenic transmission electron microscopy, dynamic light scattering, asymmetric field flow fractionation, and two-dimensional diffusion ordered spectroscopy nuclear magnetic resonance. Biphasic models to study drug transfer kinetics demonstrated that COEs with qualitative and quantitative sameness and comparable GSDs, analyzed using earth mover's distance, can be therapeutic equivalents. This body of research facilitated the review and approval of the first U.S. generic COE. In addition, the methods and fundamental understanding developed from this research may support the development and assessment of other complex generics. The approval of a generic COE should improve the availability of this complex drug product to U.S. patients.

Design, synthesis of novel benzimidazole derivatives as ENL inhibitors suppressing leukemia cells viability via downregulating the expression of MYC.

Eleven-Nineteen-Leukemia Protein (ENL) containing YEATS domain, a potential drug target, has emerged as a reader of lysine acetylation. SGC-iMLLT bearing with benzimidazole scaffold was identified as an effective ENL inhibitor, but with weak activity against mixed-lineage leukemia (MLL)-rearranged cells proliferation. In this study, a series of compounds were designed and synthesized by structural optimization on SGC-iMLLT. All the compounds have been evaluated for their ENL inhibitory activities. The results showed that compounds 13, 23 and 28 are the most potential ones with the IC50 values of 14.5 ± 3.0 nM, 10.7 ± 5.3 nM, and 15.4 ± 2.2 nM, respectively, similar with that of SGC-iMLLT. They could interact with ENL protein and strengthen its thermal stability in vitro. Among them, compound 28 with methyl phenanthridinone moiety replacement of indazole in SGC-iMLLT, exhibited significantly inhibitory activities towards MV4-11 and MOLM-13 cell lines with IC50 values of 4.8 µM and 8.3 µM, respectively, exhibiting ∼7 folds and ∼9 folds more potent inhibition of cell growth than SGC-iMLLT. It could also increase the ENL thermal stability while SGC-iMLLT had no obvious effect on leukemia cells. Moreover, compound 28 could downregulate the expression of target gene MYC either alone or in combination with JQ-1 in cells, which was more effective than SGC-iMLLT. Besides, in vivo pharmacokinetic studies showed that the PK properties for compound 28 was much improved over that of SGC-iMLLT. These observations suggested compound 28 was a potential ligand for ENL-related MLL chemotherapy.

Determining critical overlap concentration of polyethylene oxide to support excipient safety assessment of opioid products.

Intravenous administration of abuse-deterrent opioid products poses high safety risks, in part due to the presence of high molecular weight polymeric excipients. Previous in vivo studies in animal models have shown that the higher molecular weight (Mw) polymeric excipients like polyethylene oxide (PEO) were directly linked to such adverse responses as intravenous hemolysis and kidney damage. PEO polymers have been widely used in abuse-deterrent formulations (ADF) of opioid products, adding to concerns over the general safety of the opioid category due to the unknown safety risk from abuse via unintended routes. The current study focused on the determination of the critical overlap concentration (c*) at various PEO molecular weights to aid in explaining differences in observed adverse responses from previous animal studies on the intravenous administration of PEO solutions. Adverse in vivo responses may be related to the viscoelastic regime of the polymer solution, which depends not only on Mw but also on concentration. Having a localized polymer concentration in the blood above the c*, i.e., the transition from the dilute to semi-dilute entangled viscoelastic regime, may influence the flow behavior and interactions of cells in the blood. The relationship of c* to this combination of physical, chemical, and rheological effects is a possible driving force behind adverse in vivo responses.

Understanding syringeability and injectability of high molecular weight PEO solution through time-dependent force-distance profiles.

Opioid medications play a vital role in treating moderate to severe pain. Unfortunately, many drug misusers and abusers attempt to alter the formulations or properties of these drugs by manipulation, (e.g., crushing, chewing, smoking, snorting, injecting). The intravenous (IV) route is most dangerous to abusers, as the drugs directly enter the circulatory system and produce intense euphoria. To obtain a full understanding of the impact of syringe factors (e.g., needle gauge size, needle length, syringe barrel size), on the ease of injection, we undertook a comprehensive assessment of syringeability and injectability of manipulated abuse-deterrent formulations (ADFs). A texture analyzer-based testing method was developed for the measurement of the resistance force of pulling, holding, and pushing phases of injections. Results showed that the finer needle gauge sizes required higher injection force to withdraw drug solutions. In addition, the syringed liquid volume was highly dependent on needle gauge size, holding time, and sample viscosity. In most cases, a lower needle gauge number and a longer holding time increased the syringed volume. Needle length was highly correlated to injection force (R2 = 0.99). Using longer needles to inject drug solution requires greater force. Furthermore, large barrel size was correlated to pushing force (R2 = 0.99); thus, increasing the difficulty of pushing the plunger of a large syringe with one hand. Finally, relationships between injection force, sample viscosity, and testing conditions were elucidated using a mathematical model, which could be used in the future to assess and predict injection force of solution samples.

Low-dose dexamethasone promotes osteoblast viability by activating autophagy via the SGK1/FOXO3a signaling pathway.

Autophagy contributes to bone homeostasis and development under physiological conditions. Although previous studies have demonstrated the induction of the autophagy machinery by endogenous glucocorticoids (GCs), the precise mechanisms involved have not yet been clarified. The current study aimed to explore the effect of a low dose of GC (10-8 M dexamethasone, Dex) on autophagy in mouse embryonic osteoblastic precursor cells (MC3T3-E1 cells) and the potential mechanisms. The results showed that 10-8 M Dex induced significant time-dependent increases in the expression and activation of serum- and glucocorticoid-induced kinase-1 (SGK1) in MC3T3-E1 cells and that these effects were accompanied by increased cell viability and decreased apoptosis. The autophagy inhibitor 3-MA significantly inhibited Dex-mediated promotion of viability. Moreover, Dex increased LC3II and Beclin-1 levels and decreased SQSTM/p62 levels in a time-dependent manner, and these effects were attenuated by pretreatment with 3-MA. Transfection of Dex-treated MC3T3-E1 cells with shRNA-SGK1 resulted in a significant reduction in cell viability and an increase in apoptosis. 3-MA further exacerbated these effects of SGK1 inhibition. Knocking down SGK1 before Dex exposure significantly reduced the phosphorylated forkhead box O3a (p-FOXO3a)/FOXO3 ratio, suppressed LC3II and Beclin-1 levels, and increased SQSTM/p62 levels in MC3T3-E1 cells, and these effects were amplified by 3-MA. In conclusion, the results revealed that low-dose GC treatment increased osteoblast viability by activating autophagy via the SGK1/FOXO3a pathway.

Centimeter-scale perovskite SrTaO2N single crystals with enhanced photoelectrochemical performance.

Large-scale single crystals have potential applications in many fields, such as in ferroelectric and photoelectric energy conversion devices. Perovskite oxynitrides have also attracted attention in photoelectrochemical water splitting systems because of their high theoretical solar-to-hydrogen efficiencies. Nevertheless, the synthesis of perovskite oxynitride single crystals requires the coupling of cation exchange and ammonization processes, which is exceptionally challenging. The present study demonstrates an inorganic vapor method that provides, for the first time ever, high-quality epitaxial perovskite SrTaO2N single crystals on the centimeter scale. Assessments using Raman spectroscopy, crystal structure analysis and density functional theory determined that the conversion mechanism followed a topotactic transition mode. Compared with conventional SrTaO2N particle-assembled films, the SrTaO2N single crystals made in this work were free of interparticle interfaces and grain boundaries, which exhibited extremely high performance during photoelectrochemical water oxidation. In particular, these SrTaO2N single crystals showed the highest photocurrent density at 0.6 V vs. RHE (1.20 mA cm-2) and the highest photocurrent filling factor (47.6%) reported to date, together with a low onset potential (0.35 V vs. RHE). This onset potential was 200 mV less than that of the reported in situ SrTaO2N film, and the photocurrent fill factor was improved by 2 to 3 times.

Association of oral health knowledge, self-efficacy and behaviours with oral health-related quality of life in Chinese primary school children: a cross-sectional study.

OBJECTIVE: Achieving good oral health-related quality of life (OHRQOL) is of particular concern in children. The inter-relations among oral health knowledge, self-efficacy, behaviours and OHRQOL in children groups remain unclear. This study aimed to explore the inter-relations between these oral health behaviour-related factors and OHRQOL in primary school children. METHODS: In this cross-sectional study, 651 children in grades 2 and 3 were recruited in October 2020 from two primary schools in Minhang District, Shanghai, China. Data were collected through self-reported questionnaires, consisting of demographic characteristics, oral health knowledge, self-efficacy, oral health behaviours and OHRQOL. Pearson's correlation analyses were used to analyse the relationship between study variables. Structural equation models were used to test the inter-relations between OHRQOL and oral health behaviour-related factors. RESULTS: Four hypothetical structural equation models were tested and one of them was selected as the most appropriate model, which explained 15.0% of the variance in OHRQOL. This selected model showed that oral health behaviours were directly related to OHRQOL. Oral health knowledge was indirectly associated with OHRQOL through both self-efficacy and oral health behaviours. Self-efficacy was directly associated with OHRQOL or was indirectly associated with OHRQOL through oral health behaviours. CONCLUSION: This study revealed a pathway of association between children's oral health knowledge and their OHRQOL, in which children's oral health self-efficacy and behaviours had indirect effects. This provides a basis for understanding the mechanism of oral health promotion interventions to improve children's OHRQOL and helps to identify direct or indirect intervention targets.

Restoring mitochondrial cardiolipin homeostasis reduces cell death and promotes recovery after spinal cord injury.

Alterations in phospholipids have long been associated with spinal cord injury (SCI). However, their specific roles and signaling cascades in mediating cell death and tissue repair remain unclear. Here we investigated whether alterations of cardiolipin (CL), a family of mitochondrion-specific phospholipids, play a crucial role in mitochondrial dysfunction and neuronal death following SCI. Lipidomic analysis was used to determine the profile of CL alteration in the adult rat spinal cord following a moderate contusive SCI at the 10th thoracic (T10) level. Cellular, molecular, and genetic assessments were performed to determine whether CL alterations mediate mitochondrial dysfunction and neuronal death after SCI, and, if so, whether reversing CL alteration leads to neuroprotection after SCI. Using lipidomic analysis, we uncovered CL alterations at an early stage of SCI. Over 50 distinct CL species were identified, of which 50% showed significantly decreased abundance after SCI. The decreased CL species contained mainly polyunsaturated fatty acids that are highly susceptible to peroxidation. In parallel, 4-HNE, a lipid peroxidation marker, significantly increased after SCI. We found that mitochondrial oxidative stress not only induced CL oxidation, but also resulted in CL loss by activating cPLA2 to hydrolyze CL. CL alterations induced mitochondrial dysfunction and neuronal death. Remarkably, pharmacologic inhibition of CL alterations with XJB-5-131, a novel mitochondria-targeted electron and reactive oxygen species scavenger, reduced cell death, tissue damage and ameliorated motor deficits after SCI in adult rats. These findings suggest that CL alteration could be a novel mechanism that mediates injury-induced neuronal death, and a potential therapeutic target for ameliorating secondary SCI.

Effects of social support on oral health behavior: Serial multiple-mediator model.

OBJECTIVE: This study investigates the serial multiple mediation of job stress and perceived behavioral control (PBC) in the relationship between social support and oral health behavior (OHB). MATERIALS AND METHODS: This cross-sectional study was conducted in Minhang District, Shanghai, China, between July and November 2021. 559 Employees from 10 community health centers were recruited. A five-part survey was used to collect data on demographic characteristics, social support, job stress, PBC and OHB. Descriptive statistics, independent-sample t-test, Pearson's correlation analysis, ordinary least-squares regression and the bootstrap method were used to analyze data. RESULTS: Social support was found to impact OHB through three significant mediation pathways: the pathway through job stress (point estimate = 0.033; 95%BC CI [0.004, 0.089]), the pathway through PBC (point estimate = 0.283; 95%BC CI [0.174, 0.416]), and the serial multiple mediating pathway through first job stress and then PBC (point estimate = 0.006; 95%BC CI [0.001, 0.019]). CONCLUSION: This study indicates that job stress and PBC play a serial multiple-mediator role in the relationship between social support and OHB. To ensure the effectiveness of social support intervention on OHB, workplace managers and employees should strengthen the motivators that could promote the reduction of job stress and the improvement of PBC.

RabbitFX: Efficient Framework for FASTA/Q File Parsing on Modern Multi-Core Platforms.

The continuous growth of generated sequencing data leads to the development of a variety of associated bioinformatics tools. However, many of them are not able to fully exploit the resources of modern multi-core systems since they are bottlenecked by parsing files leading to slow execution times. This motivates the design of an efficient method for parsing sequencing data that can exploit the power of modern hardware, especially for modern CPUs with fast storage devices. We have developed RabbitFX, a fast, efficient, and easy-to-use framework for processing biological sequencing data on modern multi-core platforms. It can efficiently read FASTA and FASTQ files by combining a lightweight parsing method by means of an optimized formatting implementation. Furthermore, we provide user-friendly and modularized C++ APIs that can be easily integrated into applications in order to increase their file parsing speed. As proof-of-concept, we have integrated RabbitFX into three I/O-intensive applications: fastp, Ktrim, and Mash. Our evaluation shows that the inclusion of RabbitFX leads to speedups of at least 11.6 (6.6), 2.4 (2.4), and 3.7 (3.2) compared to the original versions on plain (gzip-compressed) files, respectively. These case studies demonstrate that RabbitFX can be easily integrated into a variety of NGS analysis tools to significantly reduce associated runtimes. It is open source software available at https://github.com/RabbitBio/RabbitFX.

[Carbon Sequestration Characteristics of Different Restored Vegetation Types in Loess Hilly Region].

Since 1999, the "Grain for Green" Program has been extensively implemented in the Loess Plateau region. This measure has largely been of concern not only for its contribution to soil erosion reduction but also for its effects on carbon sequestration. The aim of this study was to assess the carbon sequestration characteristics of different restored vegetation types in areas with severe soil erosion on the Loess Plateau and to compare the effects of restoration age and slope direction on the vegetation carbon sequestration. To evaluate the carbon density and composition characteristics of different ecosystem types, six typical vegetation types (including farmland, grassland, Hippophae rhamnoides Linn., Caragana korshinskii Kom., Robinia pseudoacacia L., and Populus davidiana Dode.) were selected in the Loess Hilly Region, i.e., Wuqi County and Zhidan County in Northern Shaanxi province, which is a typical area for the implementation of artificial vegetation restoration. The results showed that:① vegetation restoration in the semi-arid loess region had a profound impact on carbon sequestration. The carbon density of different vegetations, as well as different vegetation components including above-ground vegetation, below-ground roots, and litter, shared the same pattern as tree>scrub>grassland>farmland. The 0-40 cm soil layer of the farmland showed the lowest soil organic carbon density (1355.5 g·m-2), compared to which those of grassland, H. rhamnoides Linn., C. korshinskii Kom., R. pseudoacacia L., and P. davidiana Dode. were higher by 91.4%, 125.2%, 144.0%, 124.5%, and 232.6%, respectively. ② It was common in grassland, H. rhamnoides Linn., C. korshinskii Kom., and P. davidiana Dode. for the carbon density of different vegetation components as well as soil organic carbon density of different soil layers (0-5, 5-20, and 20-40 cm) to generally show an increasing trend with increased restoration age. ③ Slope direction had a significant impact on the vegetation carbon density only for H. rhamnoides Linn., C. korshinskii Kom., R. pseudoacacia L., and P. davidiana Dode., while showing the contrary for farmland and grassland. Soil organic carbon densities for sunny slopes were significantly lower than those for shaded slopes by 22.9%, 34.3%, 75.8%, 49.1%, 22.4%, and 69.4%, respectively, for farmland, grassland, H. rhamnoides Linn., C. korshinskii Kom., R. pseudoacacia L., and P. davidiana Dode. ④ Ecosystem carbon density varied significantly for different ecosystem types, among which farmland showed the lowest (2022.1 g·m-2), and grassland, H. rhamnoides Linn., C. korshinskii Kom., R. pseudoacacia L., and P. davidiana Dode. showed values higher by 48.7%, 152.8%, 125.1%, 166.3%, and 530.7%, respectively. The carbon density of each ecosystem component showed a pattern as follows:soil layer>above-ground vegetation layer>root layer>litter layer. Soil organic carbon constituted the main part of the ecosystem carbon density and accounted for 67.0%, 86.3%, 59.7%, 72.7%, and 56.5%, respectively, for farmland, grassland, H. rhamnoides Linn., C. korshinskii Kom., and R. pseudoacacia L. These results can provide an essential basis for scientific management of ecosystem carbon pools and promote ecological environment management on the Loess Plateau.

Clinical features and viral etiology of acute respiratory infection in an outpatient fever clinic during COVID-19 pandemic in a tertiary hospital in Nanjing, China.

BACKGROUND: Clinical feature and viral etiology for acute respiratory infection (ARI) in the community was unknown during coronavirus disease 2019 (COVID-19) pandemic. OBJECTIVE: In a retrospective study, we aimed to characterize the clinical feature and etiology for the ARI patients admitted to the outpatient fever clinic in Nanjing Drum Tower Hospital between November 2020 and March 2021. METHODS: Fifteen common respiratory pathogens were tested using pharyngeal swabs by multiplex reverse transcriptase-polymerase chain reaction assays. RESULTS: Of the 242 patients, 56 (23%) were tested positive for at least one viral agent. The predominant viruses included human rhinovirus (HRV) (5.4%), parainfluenza virus type III (PIV-III) (5.0%), and human coronavirus-NL63 (HCoV-NL63) (3.7%). Cough, sputum, nasal obstruction, and rhinorrhea were the most prevalent symptoms in patients with viral infection. Elderly and the patients with underlying diseases were susceptible to pneumonia accompanied with sputum and chest oppression. Three (5.4%) patients in virus infection group, whereas 31 (16.7%) in non-viral infection group (p = 0.033), were empirically prescribed with antiviral agents. Among 149 patients who received antibiotic therapy, 30 (20.1%) patients were later identified with viral infection. CONCLUSION: Our study indicated the importance of accurate diagnosis of ARI, especially during the COVID-19 pandemic, which might facilitate appropriate clinical treatment.

Integrated Analysis of the Role of Enolase 2 in Clear Cell Renal Cell Carcinoma.

Enolase 2 (ENO2) has increasingly been documented in multiple cancers in recent years. However, the role of ENO2 in clear cell renal carcinoma (ccRCC) has not been fully explored. In the present study, open-access data were downloaded from The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO), and the Human Protein Atlas (HPA) databases. All statistical analyses were performed in R and GraphPad Prism 8 softwares. Results showed that ENO2 was overexpressed in ccRCC tissues and cell lines and correlated with worse clinical features and prognosis. In vitro experiments indicated that the inhibition of ENO2 could hamper the malignant behaviors of ccRCC cells. Gene Set Enrichment Analysis showed that epithelial-mesenchymal transition, KRAS signaling, inflammatory response, angiogenesis, hypoxia, and WNT/ß-catenin pathways were upregulated in the ENO2 high-expression group; whereas adipogenesis, DNA repair, and androgen response pathways were downregulated. Immune infiltration analysis indicated that patients with high ENO2 levels might have higher M2 macrophages and lower Î³ß T cells in the tumor microenvironment, which may account to some extent for the worse prognosis of ENO2. Moreover, it was found that patients with low and high ENO2 expression might be more sensitive to PD-1 therapy and CTLA-4 therapy, respectively. In addition, patients with high ENO2 expression showed lower sensitivity to common chemotherapy drugs for ccRCC, including axitinib, cisplatin, gemcitabine, pazopanib, sunitinib, and temsirolimus. Overall, these results suggest that ENO2 is a potential prognosis biomarker of ccRCC and could affect the malignant biological behavior of cancer cells, highlighting its value as a potential therapeutic target.

Gut microbiota is associated with the effect of photoperiod on seasonal breeding in male Brandt's voles (Lasiopodomys brandtii).

BACKGROUND: Seasonal breeding in mammals has been widely recognized to be regulated by photoperiod, but the association of gut microbiota with photoperiodic regulation of seasonal breeding has never been investigated. RESULTS: In this study, we investigated the association of gut microbiota with photoperiod-induced reproduction in male Brandt's voles (Lasiopodomys brandtii) through a long-day and short-day photoperiod manipulation experiment and fecal microbiota transplantation (FMT) experiment. We found photoperiod significantly altered reproductive hormone and gene expression levels, and gut microbiota of voles. Specific gut microbes were significantly associated with the reproductive hormones and genes of voles during photoperiod acclimation. Transplantation of gut microbes into recipient voles induced similar changes in three hormones (melatonin, follicle-stimulating hormone, and luteinizing hormone) and three genes (hypothalamic Kiss-1, testicular Dio3, and Dio2/Dio3 ratio) to those in long-day and short-day photoperiod donor voles and altered circadian rhythm peaks of recipient voles. CONCLUSIONS: Our study firstly revealed the association of gut microbiota with photoperiodic regulation of seasonal breeding through the HPG axis, melatonin, and Kisspeptin/GPR54 system. Our results may have significant implications for pest control, livestock animal breeding, and human health management. Video Abstract.