Identifying differentially expressed genes in goat mammary epithelial cells induced by overexpression of SOCS3 gene using RNA sequencing.

The suppressor of cytokine signaling 3 (SOCS3) is a key signaling molecule that regulates milk synthesis in dairy livestock. However, the molecular mechanism by which SOCS3 regulates lipid synthesis in goat milk remains unclear. This study aimed to screen for key downstream genes associated with lipid synthesis regulated by SOCS3 in goat mammary epithelial cells (GMECs) using RNA sequencing (RNA-seq). Goat SOCS3 overexpression vector (PC-SOCS3) and negative control (PCDNA3.1) were transfected into GMECs. Total RNA from cells after SOCS3 overexpression was used for RNA-seq, followed by differentially expressed gene (DEG) analysis, functional enrichment analysis, and network prediction. SOCS3 overexpression significantly inhibited the synthesis of triacylglycerol, total cholesterol, non-esterified fatty acids, and accumulated lipid droplets. In total, 430 DEGs were identified, including 226 downregulated and 204 upregulated genes, following SOCS3 overexpression. Functional annotation revealed that the DEGs were mainly associated with lipid metabolism, cell proliferation, and apoptosis. We found that the lipid synthesis-related genes, STAT2 and FOXO6, were downregulated. In addition, the proliferation-related genes BCL2, MMP11, and MMP13 were upregulated, and the apoptosis-related gene CD40 was downregulated. In conclusion, six DEGs were identified as key regulators of milk lipid synthesis following SOCS3 overexpression in GMECs. Our results provide new candidate genes and insights into the molecular mechanisms involved in milk lipid synthesis regulated by SOCS3 in goats.

Excess Mortality for Femoral Intertrochanteric Fracture Patients Aged 50 Years and Older Treated Surgically and Conservatively in Tianjin, China: A Cohort Study.

OBJECTIVE: Intertrochanteric fracture is one type of hip fracture, which is the most serious consequence of osteoporosis. Along with the growing elderly population, intertrochanteric fracture is expected to rise increasingly. The aim of this study was to assess excess mortality after intertrochanteric fractures and to identify the predictors of long-term mortality by therapy among patients aged 50 years and older in Tianjin. METHODS: This is a retrospective cohort study on mortality for 3029 patients aged 50 years and older in Tianjin experiencing an intertrochanteric fracture between December 26, 2014 and December 31, 2018. Data were from Tianjin Hospital Hip Fracture (THHF) cohort. Follow-up period was until March 31, 2022. Mortality, excess mortality, and comorbidities were analyzed and stratified by therapy and gender. Time dependent Cox models were performed to estimate the effects of the variables. RESULTS: Absolute mortality for all the patients was 5.90% at 3 months, 12.55% at 12 months, 19.92% at 24 months and 27.28% at 36 months. Absolute mortality for surgical group was 1.57% at 3 months, 4.77% at 12 months, 8.49% at 24 months and 12.07% at 36 months, significantly lower than conservative group: 10.50% at 3 months, 20.73% at 12 months, 31.96% at 24 months and 43.04% at 36 months. We found a substantially lower mortality (hazard ratio [HR] 0.34, 95% confidence internal, [CI]: 0.23-0.52, p = 0.000) among patients undergoing surgical therapy than those undergoing conservative therapy, even when controlled for gender, age, the length of hospital stay, and all the comorbidities. Female patients (HR 0.68, 95% CI: 0.58-0.79, p = 0.000) were less likely to die than male patients after an intertrochanteric fracture. Patients treated by the two methods were both found to have excess mortality rates compared to the general population, although in different levels. The excess mortality rates for patients in the conservative therapy group were 14.46% in males and 17.93% in females, while in the surgical therapy group, 2.78% in females and 4.37% in males. The comorbidities moderate or severe renal disease (HR 2.19, 95% CI: 1.61-2.98, p = 0.000), metastatic solid tumor (HR 6.35, 95% CI: 1.56-25.85, p = 0.010), hypoproteinemia (HR 1.22, 95% CI: 1.01-1.47, p = 0.034), and older age (HR 1.89, 95% CI: 1.73-2.08, p = 0.000) were also risk factors on mortality. A worse-case analysis for the primary outcome were performed as sensitivity analysis and it was consistent with the original conclusion. CONCLUSION: Intertrochanteric factures for people aged 50 years older were found to have excess mortality compared to the general population in Tianjin city, and preventing the fractures in the hip for elderly people was imperative. After controlling tfor comorbidities and age, female gender and surgical therapy were protective factors for the death after fractures, which could provide strong evidence for patients and surgeons to make decisions.

Mucin1 induced trophoblast dysfunction in gestational diabetes mellitus via Wnt/ß-catenin pathway.

BACKGROUND: To elucidate the role of Mucin1 (MUC1) in the trophoblast function (glucose uptake and apoptosis) of gestational diabetes mellitus (GDM) women through the Wnt/ß-catenin pathway. METHODS: Glucose uptake was analyzed by plasma GLUT1 and GLUT4 levels with ELISA and measured by the expression of GLUT4 and INSR with immunofluorescence and Western blotting. Apoptosis was measured by the expression of Bcl-2 and Caspase3 by Western blotting and flow cytometry. Wnt/ß-catenin signaling measured by Western blotting. In vitro studies were performed using HTR-8/SVneo cells that were cultured and treated with high glucose (HG), sh-MUC1 and FH535 (inhibitor of Wnt/ß-catenin signaling). RESULTS: MUC1 was highly expressed in the placental trophoblasts of GDM, and the Wnt/ß-catenin pathway was activated, along with dysfunction of glucose uptake and apoptosis. MUC1 knockdown resulted in increased invasiveness and decreased apoptosis in trophoblast cells. The initial linkage between MUC1, the Wnt/ß-catenin pathway, and glucose uptake was confirmed by using an HG-exposed HTR-8/SVneo cell model with MUC1 knockdown. MUC1 knockdown inhibited the Wnt/ß-catenin signaling pathway and reversed glucose uptake dysfunction and apoptosis in HG-induced HTR-8/SVneo cells. Meanwhile, inhibition of Wnt/ß-catenin signaling could also reverse the dysfunction of glucose uptake and apoptosis. CONCLUSIONS: In summary, the increased level of MUC1 in GDM could abnormally activate the Wnt/ß-catenin signaling pathway, leading to trophoblast dysfunction, which may impair glucose uptake and induce apoptosis in placental tissues of GDM women.

Ga-Modification Near-Surface Composition of Pt-Ga/C Catalyst Facilitates High-Efficiency Electrochemical Ethanol Oxidation through a C2 Intermediate.

In electrochemical ethanol oxidation reactions (EOR) catalyzed by Pt metal nanoparticles through a C2 route, the dissociation of the C-C bond in the ethanol molecule can be a limiting factor. Complete EOR processes producing CO2 were always exemplified by the oxidative dehydrogenation of C1 intermediates, a reaction route with less energy utilization efficiency. Here, we report a Pt3Ga/C electrocatalyst with a uniform distribution of Ga over the nanoparticle surface for EOR that produces CO2 at medium potentials (>0.3 V vs SCE) efficiently through direct and sustainable oxidation of C2 intermediate species, i.e., acetaldehyde. We demonstrate the excellent performance of the Pt3Ga-200/C catalyst by using electrochemical in situ Fourier transform infrared reflection spectroscopy (FTIR) and an isotopic labeling method. The atomic interval structure between Pt and Ga makes the surface of nanoparticles nonensembled, avoiding the formation of poisonous *CHx and *CO species via bridge-type adsorption of ethanol molecules. Meanwhile, the electron redistribution from Ga to Pt diminishes the *O/*OH adsorption and CO poisoning on Pt atoms, exposing more available sites for interaction with the C2 intermediates. Furthermore, the dissociation of H2O into *OH is facilitated by the high hydrophilicity of Ga, which is supported by DFT calculations, promoting the deep oxidation of C2 intermediates. Our work represents an extremely rare EOR process that produces CO2 without observing kinetic limitations under medium potential conditions.

Chromatographic methods for rapid aflatoxin B1 analysis in food: a review.

Aflatoxin B1 (AFB1) is a mycotoxin and is the most carcinogenic of all known chemicals. In view of the AFB1 characteristics of widespread distribution, serious pollution, great harm to humans, and animals and difficult to remove, it is urgent to develop a convenient and sensitive detection method. Moreover, chromatographic test strips (CTSs) are a rapid detection technology that combines labeling technology with chromatography technology. CTSs have been widely used in the fields of environmental monitoring, medical diagnosis, and food safety analysis in recent years. Different from other immune assays, they have the advantages of short measuring time, low cost, high efficiency and no need for professionals to operate. In addition, the introduction of nanomaterials has laid a good foundation for the detection of high sensitivity, high specificity and high efficiency via CTSs. Herein, we tend to comprehensively introduce the applications of chromatographic methods in AFB1 detection and pay attention to the signal detection modes based on nanomaterials in antibody-based immunochromatographic strips (ICSs), such as colorimetric, fluorescent, chemiluminescent, and Raman scattering sensing. Some typical examples are also listed in this review. In the end, we make a summary and put forward prospects for the development of CTSs.

This review is the first systematic review about the applications of antibody-/aptamer-based chromatographic methods for rapid AFB1 detection.Pay attention to the signal detection modes based on nanomaterials in antibody-ICSs, such as colorimetric, fluorescent, chemiluminescent and Raman scattering sensing.Make a summary about some typical examples and put forward prospects for the development of CTSs.

SPECT/CT imaging features of cystic degeneration of the talus and their relation to pathological findings.

BACKGROUND: Osteochondral lesions of the talus (OLTs) are a common orthopedic condition. The image presentation is very similar to that of ischemic necrosis of the talus complicated by a talar neck fracture, but the two are very different lesions. When abnormalities in bone density (or signal) of the talar body (apex of the fornix) with concomitant bone defects and cystic changes are found on X-ray, computed tomography (CT), or magnetic resonance imaging, it is important to accurately determine the nature of the lesion and make a correct diagnosis for the treatment and prognosis of the patient. The purpose of this study was to explore the imaging features of three-phase single-photon emission computed tomography (SPECT)/CT images of cystic lesions of the talus. METHODS: A total of 189 patients with chronic pain in the ankle joint suspected to be caused by cystic degeneration of the talus were enrolled. All patients underwent 99mTc-methyl diphosphonate (99mTc-MDP) three-phase SPECT/CT bone imaging and delayed scans in our hospital. The location, range of involvement, classification, CT value, and radioactivity uptake of the sclerotic areas of cystic lesions on the talus, and the continuity of the articular surface, were recorded. All recorded parameters were analyzed in comparison with pathological results. RESULTS: Eighty-three percent (157/189) of the talar cysts were located on the medial fornix, largely involving the anterior middle part (43.27%), with larger cysts involving the posterior part (9.6%). Sixty-three percent (119/189) of the patients had type I lesions and 37% (70/189) had type II lesions. The articular surface of the medial dome of the talus was intact in all patients, but the subchondral bony articular surface was rough in 88% (166/189) of patients. The coincidence rate for the location, type, and range of involvement of cystic lesions with the pathological results was 87.83% (166/189). The mean CT value of the cystic lesions was 45 ± 15 HU (30-60 HU). The percentages of pathological chondrogenesis in high CT value ≥ 50 HU (19/70) and low CT value < 50 HU (51/70) groups were 89.47% (17/19) and 29.14% (15/51) (χ2 = 20.12, p < 0.001), respectively. The target/background ratio (T/B ratio) of the radioactivity-uptake area of the talus vault was 2.0 ± 0.5 (1.5-2.5). The percentages of pathological new trabecular bone in those with a T/B ratio ≥ 2.0 (157/189) and T/B ratio < 2.0 (32/189) were 82.80% (130/157) and 25.00% (8/32; χ2 = 45.08, p < 0.001), respectively. CONCLUSIONS: Three-phase bone imaging could identify damage of the talus caused by cystic degeneration, while delayed SPECT/CT images showed advantages for displaying bone microstructure, blood supplement, and bone metabolism when examining the location, range of involvement, classification, and repair of cystic lesions of the talus.

Does completion of radical hysterectomy improve oncological outcomes of women with clinical early-stage cervical cancer and intraoperative detection of nodal involvement?: protocol for a systematic review and meta-analysis.

INTRODUCTION: The management of women with clinical early-stage cervical cancer and lymph node involvement detected intraoperatively is heterogeneous and controversial. This paper presents the protocol of a systematic review and meta-analysis regarding the management of this specific population of patients. This proposed study aims to answer the question: does completion of radical hysterectomy improve the oncological outcomes of women with clinical early-stage cervical cancer and intraoperatively detected nodal involvement? METHODS AND ANALYSIS: This protocol is drafted following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Protocols guidelines, and the proposed study will be conducted in accordance with the standard guidelines of 'Preferred Reporting Items for Systematic Reviews and Meta-Analyses' and 'Meta-analysis of Observational Studies in Epidemiology reporting guideline'. Comprehensive literature searches will be performed in PubMed, Embase, Scopus, and Web of Science. The screening of the eligible studies, the extraction of data of interest, and the quality assessment of the included studies will all be independently performed by different members of our team. The primary outcome of this proposed study will be comparing the risk of recurrence or death from cervical cancer and the risk of all-cause death in patients with two different treatments (completion of radical hysterectomy or abandonment of radical hysterectomy); the secondary outcome of this proposed study will be comparing the risk of the grade 3/4 toxicities associated with the two types of management. Given the clinical heterogeneity among the included studies, data on outcomes will be pooled by random-effects models. Heterogeneity will be evaluated using the I2 statistic. The risk of bias for the included studies will be evaluated using the Newcastle-Ottawa Scale or the Cochrane collaboration's tool. The grade of evidence will be evaluated by two independent members of our team using the Grading of Recommendations, Assessment, Development and Evaluations approach. ETHICS AND DISSEMINATION: Ethical approval is not required because there will no primary data collected. The findings of this proposed study will be published in an international peer-reviewed journal. PROSPERO REGISTRATION NUMBER: CRD42021273527.

Design of a Signal-Amplified Aptamer-Based Lateral Flow Test Strip for the Rapid Detection of Ochratoxin A in Red Wine.

In order to improve the weak optical performance of gold nanoparticles and realize the signal amplification of lateral flow chromatography test strips, individual gold nanoparticles (AuNPs) were aggregated into gold nanoparticle aggregates through functional groups around polyamidoamine (PAMAM) dendrimers. A signal-amplified aptamer-based lateral flow chromatography test strip was constructed for the rapid determination of ochratoxin A (OTA). Under optimal conditions, the visual detection limit of this test strip was 0.4 ng mL-1 and the semi-quantitative limit of detection (LOD) was 0.04 ng mL-1. Compared with other traditional aptamer lateral flow chromatography test strips, its sensitivity was improved about five times. The whole test could be completed within 15 min. The aptamer-based strip was applied to the detection of OTA in red wine; the average recoveries ranged from 93% to 105.8% with the relative standard deviation (RSD) varying from 3% to 8%, indicating that the test strip may be a potentially effective tool for the on-site detection of OTA.

Incidence of omental metastasis in uterine serous carcinoma: a systematic review and meta-analysis.

OBJECTIVES: Previous studies examining the incidence of omental metastasis in uterine serous carcinoma generally suffered from small sample size, retrospective observational design, and single-center setting. So far, there was no systematic review and meta-analysis available on this topic, we conducted this study to quantitatively synthesize the data relating to this topic. DESIGN: systematic review and meta-analysis. MATERIAL AND METHODS: PubMed, Embase, and Web of Science were searched up until August 15, 2020. Two reviewers independently performed study selection, data extraction, and quality assessment. I2 was employed to assess the heterogeneity among the included studies. Effect sizes along with 95% confidence intervals were calculated to analyze outcomes of interest. Funnel plots and the Egger test were used to detect the risk of publication bias. OUTCOME MEASURES: incidence of omental metastasis in uterine serous carcinoma. RESULTS: A total of 16 studies involving 1012 women with uterine serous carcinoma were included in this study. All the included studies were at low risk of bias, and the heterogeneity among them was low. The pooled incidence of overall omental metastasis, occult omental metastasis, and gross omental metastasis in uterine serous carcinoma were 18% (95% CI, 0.15-0.20), 6% (95% CI, 0.04-0.08), and 10% (95% CI, 0.08-0.13), respectively. CONCLUSIONS: Uterine serous carcinoma has a high tendency of omental metastasis. The main form of omentum involvement is gross metastasis. However, occult metastasis in the normal-looking omentum is also worthy of note.

Dissection of the potential anti-diabetes mechanism of salvianolic acid B by metabolite profiling and network pharmacology.

RATIONALE: Salvianolic acid B (Sal B), the Q-marker in Salvia miltiorrhiza, was proved to present an obvious anti-diabetes effect when treated as a food intake. Until now, the metabolism feature, tissue distribution and anti-diabetes mechanism of Sal B have not been fully elucidated. METHODS: The metabolites of Sal B in rats were profiled using ultrahigh-performance liquid chromatography coupled with time-of-flight mass spectrometry. The potential anti-diabetes mechanism of Sal B was predicted by network pharmacology. RESULTS: A total of 31 metabolites were characterized in rats after ingestion of Sal B at a dosage of 40 mg/kg, including 1 in plasma, 19 in urine, 31 in feces, 0 in heart, 0 in liver, 0 in spleen, 1 in lung, 1 in kidney and 0 in brain. Among them, 18 metabolites were reported for the first time. Phase I reactions of hydrolysis, hydrogenation, dehydroxylation, hydroxylation, decarboxylation and isomerization, and phase II reactions of methylation were found in Sal B. Notably, decarboxylation and dehydroxylation were revealed in Sal B for the first time. The pharmacology network results showed that Sal B and its metabolites could regulate ALB, PLG, ACE, CASP3, MMP9, MMP2, MTOR, etc. The above targets were involved in insulin signaling pathway, PI3K-Akt signaling pathway, HIF-1 signaling pathway, TNF signaling pathway, etc. CONCLUSIONS: The metabolism feature of Sal B in vivo was systematically revealed, and its anti-diabetes mechanism for further pharmacological validations was predicted based on metabolite profiling and network pharmacology for the first time.

Visible Light Driven and Copper-Catalyzed C(sp3)-H Functionalization of O-Pentafluorobenzoyl Ketone Oximes.

The C(sp3)-H functionalization of O-pentafluorobenzoyl ketone oximes was implemented under visible light irradiation with copper complexes as catalysts. The reactions involve iminyl-radical-mediated intramolecular hydrogen atom transfer as the key step, with the iminyl radicals being generated via copper-effected N-O cleavage. The reaction afforded 3,4-dihydro-2H-pyrroles under the conditions of [Cu(DPEphos)(bcp)]PF6 and DABCO, while γ-pentafluorobenzoyloxy ketones were produced predominantly when [Cu(dpp)2]PF6 and InCl3·4H2O were used as catalysts.

Characterization of metabolic fate of phellodendrine and its potential pharmacological mechanism against diabetes mellitus by ultra-high-performance liquid chromatography-coupled time-of-flight mass spectrometry and network pharmacology.

RATIONALE: Characterizing the functional mechanism of quality control marker (Q-marker) was of great importance in revealing the primary pharmacological mechanism of herbs or the other complex system, and drug-related metabolites always contribute to the pharmacological functions. Cortex Phellodendri was used as a core herb in the treatment of diabetes mellitus (DM). As a Q-marker of Cortex Phellodendri, the role of phellodendrine in DM was still unclear. Thus, the characterization of phellodendrine-related metabolites in vivo and the subsequent induced functional mechanism exerted great importance in elucidating the anti-DM mechanism of Cortex Phellodendri. METHODS: An ultra-high-performance liquid chromatography-coupled time-of-flight mass spectrometry (UHPLC/Q-TOF MS) method was developed to profile metabolites of phellodendrine in rats. The potential pharmacological mechanism against DM was predicted by network pharmacology. RESULTS: A total of 19 phellodendrine-related metabolites were screened out in rats for the first time. Among them, M4, M5, M9, and M12 were regarded as the primary metabolites. Meanwhile, phase I metabolic reactions of hydroxylation, demethylation, and isomerization and phase II reactions of glucuronidation and sulfation occurred to phellodendrine; glucuronidation and hydroxylation were the two main metabolic reactions. Moreover, the potential targets of phellodendrine and three main metabolites (M4, M5, and M12) were predicted by a network pharmacological method, and they mainly shared 52 targets, including PDE5A, CHRNA3, SIGMAR1, F3, ESR1, DRD1, DRD2, DRD3, and DRD4. Furthermore, Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that calcium signaling pathway, cGMP-PKG signaling pathway, and cAMP signaling pathway were regarded as the core mechanism of phellodendrine to treat DM. CONCLUSION: The metabolic feature of phellodendrine in vivo was revealed for the first time, and its anti-DM mechanism information for further pharmacological validations was also supplied. It also gave a direction to further elucidation of pharmacological mechanism of Cortex Phellodendri in treating DM.

Dissection of the potential pharmacological function of neohesperidin dihydrochalcone - a food additive - by in vivo substances profiling and network pharmacology.

Food additives are widely used in our daily life, and the side-effects caused by them have gained extensive attention around the world. Notably, constituent-oriented metabolites, in some sense, always contribute to pharmacological changes, inducing toxicity, therapeutic effects, etc. Characterization of the metabolites and their potential functions is of great importance to the practical applications. In this work, an integrated strategy by combining metabolite profiling and network pharmacology was applied to characterize the metabolic features and reveal pharmacological changes of neohesperidin dihydrochalcone (NHDC) in vivo to demonstrate its pharmacological mechanism and potential functions. As a result, a total of 19 metabolites (3 in plasma, 19 in urine, 8 in feces, 3 in heart, 5 in liver, 0 in spleen, 1 in lung, 2 in kidneys and 2 in brain) were screened and 18 of them were characterized for the first time. Phase I metabolic reactions of hydrolysis and phase II reactions of glucuronidation, sulfation, glutamylation, N-butyryl glycylation and lactylation were the main metabolic reactions of NHDC in vivo. Moreover, the results analyzed by network pharmacology revealed that, in addition to common pathways (steroid hormone biosynthesis) of NHDC, metabolites' targets were involved in pathways in cancer, ovarian steroidogenesis, proteoglycans in cancer, PI3K-Akt signaling pathway and progesterone-mediated oocyte maturation, indicating that these functional changes might result in potential novel functions or other side-effects, such as a disorder of steroid hormones. Our work provided the metabolic features and functional modifications of NHDC in vivo for the first time, and meaningful information for further pharmacological validations or potential functions is supplied.

Blood biomarkers related to osteonecrosis of femoral head by internal fixation after Garden I femoral neck fracture: a cohort study.

BACKGROUND: Internal fixation is currently considered the gold standard in treatment for femoral neck fractures in adults. However, osteonecrosis of the femoral head (ONFH) after internal fixation would occur in quite proportion of patients with femoral neck fracture, even in Garden I femoral neck fracture. The purpose of this study was to determine the association between the blood biomarkers (serum albumin, pre-albumin, total protein and total lymphocyte count) and ONFH following internal fixation of Garden I femoral neck fracture in adults. METHOD: This is a single center cohort study, in which each patient who sustained a Garden I femoral neck fracture had been treated with internal fixation, and had adequate preoperative blood examinations. The serum albumin was categorized as ≥ 40g/L or < 40g/L. The pre-albumin was categorized as ≥ 22mg/dL or < 22mg/dL. The total protein was categorized as ≥ 65g/L or < 65 g/L. The total lymphocyte count was categorized as ≥1.1× 109 /L or <1.1×109 /L. Multivariate cox proportional hazards analysis was used to assess the association between blood markers and the osteonecrosis of femoral head during the 2-years follow-up period controlling the confounders. RESULT: A total of 10 cases of ONFH were identified. Multivariate Cox regression analysis revealed that low total lymphocyte count and hypertension state were significant independent risk factors for ONFH after internal fixation for Garden I femoral head fractures. CONCLUSION: Blood biomarkers were potential predictors for ONFH after internal fixation Garden I femoral neck fractures. We suggest that routine laboratory tests might can be used to assist surgeons to identify patients at great risk of ONFH.

Dissection of the potential anti-influenza materials and mechanism of Lonicerae japonicae flos based on in vivo substances profiling and network pharmacology.

Lonicerae japonicae flos.(LJF) was widely used as a drug to treat upper respiratory tract infection or a tea to clear heat in Asian countries for thousands of years. Despite of its curative effects confirmed by modern pharmacological methods, its functional materials and mechanism against influenza were still unclear and needed further investigation. In this study, an integrated strategy based on in vivo substances profiling and network pharmacology was proposed and applied to screen out the potential anti-influenza substances and mechanism of LJF. An UHPLC/Q-TOF MS method was utilized to profile the chemical components in LJF and their metabolites in rats. The targets of absorbed prototypes were predicted by Swiss Target Prediction, and they were further analyzed by String and Kyoto Encyclopedia of Genes and Genomes (KEGG). As a result, a total of 126 chemical components mainly featuring three chemical structure types were characterized, including 70 iridoid glycosides, 17 caffeoylquinic acids, 24 flavonoids, and 15 other types compounds. Among them, ten N-contained iridoid glycosides were characterized as potential novel compounds. Moreover, 141 xenobiotics (74 prototypes and 67 metabolites) were clearly screened out in rat plasma and urine after ingestion of LJF. Phase II reactions (sulfation, glucuronidation, methylation) and phase I reactions (dehydroxylation, hydrogenation, hydrolysis, N-heterocyclization) were the main metabolic reactions of LJF in rats. Further, a total of 338 targets were predicted and TNF, PTGS2 and EGFR were the three main targets involved in the pathology of influenza. In addition to normal NF-κB pathway, T cell signal pathway and mTOR signal pathway were the other patterns for LJF to achieve its anti-flu effects. Our work provided the meaningful data for further pharmacological validation of LJF against influenza, and a new strategy was also proposed for minimizing the process to reveal the mechanism and functional basis of TCMs.

An integrated strategy for revealing the pharmacological changes based on metabolites profiling and network pharmacology: Arctiin as an example.

Traditional Chinese medicine was widely used in China since its definite effects and therapy. The components of TCM were absorbed into the circle system as the format of prototypes or metabolites, which contributed to the therapy or side effects. Declaring the functional changes in this process was of great importance to the clinical applications. In this work, an integrated strategy based on metabolites' profiling and network pharmacology was proposed for exploring the pharmacological changes of compounds in vivo. Arctiin, the main component in Fructus Arctii with various kinds of bioactivities, was used as an example. An ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry and metabolynx™software was applied to characterize the metabolites of arctiin in rats at a dosage of 100 mg/kg; network pharmacology was applied to characterize the functional changes. As a result, fifty-three metabolites (32 in plasma, 40 in urine, 19 in bile, 20 in feces, 1 in brain, 12 in liver and 4 in lungs) were screened out and characterized, and 3 of them were unambitiously identified by comparison with standard substances. Among them, 38 metabolites were reported for the first time. It was found the major metabolic pathways of arctiin in rats were demethylation, lactone-opening and phase II conjugations with sulfate and glucuronide.It also confirmed that M14, M15, M18, M23, M22, M43 and M45 were the major circulating forms of arctiin in rats following oral administration. In addition to the above metabolic reactions, phase I reactions of hydrolysis, demethylation, dehydroxylation were also observed, and dehydrogenation were first revealed metabolic patterns of arctiin in rats. Meanwhile, in addition to the main targets of arctiin (MTOR, EGFR and MAPK14), its metabolites targeted additional 392 targets with additional functions of anti-hepatitis B or viral carcinogenesis (SRC, CAPS3, PIK3CA, CDK4, ESR1, MMP9 and ERBB2). The above results provided very important information for understanding the metabolism and functional changes of arctiinin vivo, and supporting data for further pharmacological evaluation. Our work also provided a newsight for elucidation of functional changes of TCMs in vivo.

An integrated strategy for profiling the chemical components of Scutellariae Radix and their exogenous substances in rats by ultra-high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry.

RATIONALE: Traditional Chinese medicines (TCMs) attract worldwide attention because of their effects in clinical application recorded in China historical ancient codes and in records, such as 'Treatise on Febrile Diseases'. With the developments of drug analysis and research, evaluating the in vivo substances in TCMs has become of great importance. Scutellariae Radix (SR, named as huang-qing in China), the root of Scutellaria baicalensis Georgi, has shown favorable clinical effects and safety in the treatment of infection diseases; however, its in vivo compounds are unclear and need detailed investigation. METHODS: An ultra-high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC/QTOF MS) method coupled to an integrated strategy involving diagnostic ions, neutral losses and a prediction platform was used to explore the constituents of SR, and their exogenous substances in rats. RESULTS: A total of 118 chemical constituents mainly featuring five chemical structure types (flavone C-glycosides, flavone O-glycosides, free flavones, flavanones and phenylethanoid glycosides) were identified or tentatively characterized in SR, and 175 xenobiotics (68 prototypes and 107 metabolites) were profiled in rat plasma, urine, bile and feces after ingestion of SR. The metabolites were classified into four related chemical groups: flavone C-glycosides, flavone O-glycosides, flavanones and phenylethanoid glycosides. Phase II metabolism reactions, such as glucuronidation and sulfation, were the major metabolic reactions in addition to phase I reactions of hydrolysis and hydrogenation. The corresponding main metabolic features of SR in rats were also elucidated. CONCLUSIONS: The metabolism of SR, as a whole, was systemically revealed for the first time, and our work also provided meaningful information for pharmacokinetics studies and pharmacological analysis of SR in future work.

A combination of metabolite profiling and network pharmacology to explore the potential pharmacological changes of secoisolariciresinol-diglycoside.

The prototypes and metabolites formed from the use of traditional Chinese medicines (TCM) are typically the cause of both side side-effects and therapeutic results. Therefore, the characterization of in vivo substances and the determination of functional changes are of great importance for clinical applications. Secoisolariciresinol-diglycoside (SDG), one major compound in flaxseeds, was used as a potential drug to treat tumors in the clinic; however, the metabolism information and functional changes of SDG in vivo were limited, which limited its application. In this study, an integrated strategy based on metabolite profiling and network pharmacology was applied to explore the metabolism feature and functional changes of SDG. As a result, a total of 28 metabolites were found in rats, including 14 in plasma, 22 in urine, 20 in feces, 7 in the heart, 14 in the liver, 8 in the spleen, 10 in the lungs, 14 in the kidneys, and 4 in the brain. Among them, M8, M13 and M26 were the main metabolites of SDG in rats and 24 were characterized for the first time. The metabolic reactions contained phase I reactions of demethylation, dehydroxylation, deglycosylation, arabinosylation and glycosylation, and phase II reactions of glucuronidation and sulfation were also observed. Notably, the arabinosylation and glycosylation were found in SDG for the first time. Meanwhile, 121 targets of SDG and its metabolites were found, PRKCB was the main target of SDG, and the metabolites of SDG mainly targeted HSP90A1, IL6, AKT1, MAPK3, MTOR, PIK3CA, SRC, ESR1, AR, PIK3CB, and PIK3CB. The difference of targets between SDG and its metabolites could result in its additional functional pathways of neurotrophin signaling pathway, PI3K-Akt signaling pathway, HIF-1 signaling pathway or indications of anti-prostate cancer. This work provided a new insight for exploring the mechanism and therapy indications of drugs.

Visible Light-Driven Azidation/Difunctionalization of Vinyl Arenes with Azidobenziodoxole under Copper Catalysis.

Visible light-driven azidation of vinyl arenes with azidobenziodoxole as the azidating agent was investigated in acetonitrile by using Cu(I)(phenanthroline)2 complex [Cu(dap)2]PF6 as photocatalyst. The reactions generated three types of difunctionalization products, which correspond to reaction patterns of amido-azidation, benzoyloxy-azidation, and diazidation. The electronic nature of the aryl group attached to the olefin moiety was found to play a crucial role in determining the reaction consequence: when the aryl group was electron-rich, the reactions afforded benzoyloxy-azidation products exclusively; for highly electron-deficient vinyl arenes, by contrast, diazidation products were generated in moderate yields. When the aryl group was moderately electron-rich or electron-deficient, on the other hand, a three-component reaction involving acetonitrile as well as azidobenziodoxole took place to give predominantly amido-azidation products. A plausible mechanism is proposed based on the mechanistic studies to rationalize these results. The reactions of electronically less biased vinyl arenes probably proceed via a redox catalysis pathway, while the electron-rich alkenes are believed to be converted through a radical chain process. The present reactions may be of synthetic usefulness as they provide a new means for the amido-azidation of vinyl arenes.

Utility of technetium-99m-methylene diphosphonate single-photon emission computed tomography/computed tomography fusion in detecting post-traumatic chronic-infected nonunion in the lower limb.

BACKGROUND: This study aimed to evaluate fused images of single-photon emission computed tomography/computed tomography (SPECT/CT), stand-alone whole-body scintigraphy (WBS) and stand-alone CT in the diagnosis of post-traumatic chronic-infected nonunion osteomyelitis (OST) of the lower limb. PATIENTS AND METHODS: The imaging data from 144 patients with known/suspected chronic-infected fracture nonunion in the lower limbs following internal/external fixation between June 2015 and December 2017 were reviewed retrospectively. Technetium-99m-methylene diphosphonate SPECT/CT scans were performed on the patients. For each patient, the diagnosis on the basis of each imaging approach was classified as yes (OST), no (no OST), or equivocal by experienced nuclear medicine physicians and radiologists. An intraoperative bacterial culture experiment was conducted as our gold standard. The diagnostic sensitivity, specificity, accuracy, positive predictive value, negative predictive value, κ coefficient, significance level, and agreement level were analyzed. RESULTS: The diagnosis on the basis of SPECT/CT fused images showed a sensitivity of 91.3%, a specificity of 84.6%, and accuracy of 88.9% compared to that based on WBS, with a sensitivity of 52.2%, a specificity of 15.4%, accuracy of 38.9%, and CT, with a sensitivity of 65.2%, a specificity of 23.1%, accuracy of 50.0%. The fused images can show the precise sites of post-traumatic chronic-infected OST. Considerable agreement (κ 0.679) was found between the SPECT/CT diagnosis and an intraoperative bacterial culture test (WBS, κ 0.218; CT, κ = 0.184). CONCLUSION: Technetium-99m-methylene diphosphonate SPECT/CT imaging fusion can improve diagnostic confidence for post-traumatic patients with chronic nonunion OST. This imaging approach can achieve an accurate diagnosis by revealing the precise location and scope of OST with high sensitivity and specificity, which has important implications for surgical guidance by providing the precise location of OST.