Cell-free DNA testing for early hepatocellular carcinoma surveillance.

BACKGROUND: Liver cirrhosis (LC) is the highest risk factor for hepatocellular carcinoma (HCC) development worldwide. The efficacy of the guideline-recommended surveillance methods for patients with LC remains unpromising. METHODS: A total of 4367 LCs not previously known to have HCC and 510 HCCs from 16 hospitals across 11 provinces of China were recruited in this multi-center, large-scale, cross-sectional study. Participants were divided into Stage Ⅰ cohort (510 HCCs and 2074 LCs) and Stage Ⅱ cohort (2293 LCs) according to their enrollment time and underwent Tri-phasic CT/enhanced MRI, US, AFP, and cell-free DNA (cfDNA). A screening model called PreCar Score was established based on five features of cfDNA using Stage Ⅰ cohort. Surveillance performance of PreCar Score alone or in combination with US/AFP was evaluated in Stage Ⅱ cohort. FINDINGS: PreCar Score showed a significantly higher sensitivity for the detection of early/very early HCC (Barcelona stage A/0) in contrast to US (sensitivity of 51.32% [95% CI: 39.66%-62.84%] at 95.53% [95% CI: 94.62%-96.38%] specificity for PreCar Score; sensitivity of 23.68% [95% CI: 14.99%-35.07%] at 99.37% [95% CI: 98.91%-99.64%] specificity for US) (P < 0.01, Fisher's exact test). PreCar Score plus US further achieved a higher sensitivity of 60.53% at 95.08% specificity for early/very early HCC screening. INTERPRETATION: Our study developed and validated a cfDNA-based screening tool (PreCar Score) for HCC in cohorts at high risk. The combination of PreCar Score and US can serve as a promising and practical strategy for routine HCC care. FUNDING: A full list of funding bodies that contributed to this study can be found in Acknowledgments section.

[Component identification and analysis in vivo of Sanhan Huashi formula].

Sanhan Huashi formula(SHF) is the intermediate of a newly approved traditional Chinese medicine(TCM) Sanhan Huashi Granules for the treatment of COVID-19 infection. The chemical composition of SHF is complex since it contains 20 single herbal medicines. In this study, UHPLC-Orbitrap Exploris 240 was used to identify the chemical components in SHF and in rat plasma, lung and feces after oral administration of SHF, and heat map was plotted for characterizing the distribution of the chemical components. Chromatographic separation was conducted on a Waters ACQUITY UPLC BEH C_(18)(2.1 mm×100 mm, 1.7 µm) using 0.1% formic acid(A)-acetonitrile(B) as mobile phases in a gradient elution. Electrospray ionization(ESI) source was used to acquire data in positive and negative mode. By reference to quasi-molecular ions and MS/MS fragment ions and in combination with MS spectra of reference substances and compound information in literature reports, 80 components were identified in SHF, including 14 flavonoids, 13 coumarins, 5 lignans, 12 amino-compounds, 6 terpenes and 30 other compounds; 40 chemical components were identified in rat plasma, 27 in lung and 56 in feces. Component identification and characterization of SHF in vitro and in vivo lay foundations for disclosure of its pharmacodynamic substances and elucidation of the scientific connotation.

Influence of RRM, RGG and Potential Phosphorylated Sites in Cold-Inducible Protein RBM3 on its Subcellular Localization and Neuroprotective Effects.

BACKGROUND: As a potent mediator of hypothermic neuroprotection, the cold-inducible protein RBM3 is characterized with one RNA-recognition motifs (RRM) and one arginine-glycine-rich (RGG) domain. It is known that these conserved domains are required for nuclear localization in some RNA-binding proteins. However, little is known about the actual role of RRM and RGG domains in subcellular localization of RBM3. METHODS: To clarify it, various mutants of human Rbm3 gene were constructed. Plasmids were transfected into cells and the localization of RBM3 protein and its varias mutants in cells and role in neuroprotection. RESULTS: In human neuroblastoma SH-SY5Y cells, either a truncation of RRM domain (aa 1-86) or RGG domain (aa 87-157) led to an obvious cytoplasmic distribution, compared to a predominant nuclear localization of whole RBM3 protein (aa 1-157). In contrast, mutants in several potential phosphorylated sites of RBM3, including Ser102, Tyr129, Ser147, and Tyr155, did not alter the nuclear localization of RBM3. Similarly, mutants in two Di-RGG motif sites also did not affect the subcellular distribution of RBM3. Lastly, the role of Di-RGG motif in RGG domains was further investigated. The mutant of double arginines in either Di-RGG motif-1 (Arg87/90) or -2 (Arg99/105) exhibited a higher cytoplasmic localization, indicating that both Di-RGG motifs are required for nucleic localization of RBM3. CONCLUSIONS: Our data suggest that RRM and RGG domains are both required for the nuclear localization of RBM3, with two Di-RGG domain being crucial for nucleocytoplasmic shuttling of RBM3.

The development of a cSMART-based integrated model for hepatocellular carcinoma diagnosis.

BACKGROUND: Hepatocellular carcinoma (HCC) generally arises from a background of liver cirrhosis (LC). Patients with cirrhosis and suspected HCC are recommended to undergo serum biomarker tests and imaging diagnostic evaluation. However, the performance of routine diagnostic methods in detecting early HCC remains unpromising. METHODS: Here, we conducted a large-scale, multicenter study of 1675 participants including 490 healthy controls, 577 LC patients, and 608 HCC patients from nine clinical centers across nine provinces of China, profiled gene mutation signatures of cell-free DNA (cfDNA) using Circulating Single-Molecule Amplification and Resequencing Technology (cSMART) through detecting 931 mutation sites across 21 genes. RESULTS: An integrated diagnostic model called "Combined method" was developed by combining three mutation sites and three serum biomarkers. Combined method outperformed AFP in the diagnosis of HCC, especially early HCC, with sensitivities of 81.25% for all stages and 66.67% for early HCC, respectively. Importantly, the integrated model exhibited high accuracy in differentiating AFP-negative, AFP-L3-negative, and PIVKA-II-negative HCCs from LCs.

Association between Sleep and Suicidal Ideation in Chinese Undergraduate Students.

Suicide is an important global public health issue, which deserves more attention. This study aims to examine the relative independent relationship between suicide ideation and subjective sleep quality, sleep hygiene, and insomnia symptoms in undergraduate students in China. This population-based study included 2379 undergraduate students aged 18-26, randomly recruited from three public universities in Shanghai. The participants completed four questionnaires: the Pittsburgh Sleep Quality Index; Sleep Hygiene Practice Scale; Insomnia Severity Index; and the Symptom Checklist 90 (specifically the depression and anxiety dimensions and Q15-suicide ideation). The results of Spearman's correlation analysis indicate that poor sleep quality, short sleep duration, poor sleep hygiene, and insomnia symptoms were all associated with suicidal ideation in undergraduate students. However, according to the results of the hierarchical linear regression, no experience of sharing a bedroom at home, poor relationship with roommates, short sleep duration, sleep medicine use, and good daytime function were related to suicidal ideation, after controlling for the symptoms of depression and anxiety, which may be important in the identification of suicidal ideation. Sleep problems are highly discoverable and modifiable, and have a low sense of shame, therefore, sleep interventions for individuals with suicidal ideation and poor sleep quality may be an efficient and effective approach to suicide prevention.

Long-Term Prognostic Factors in Patients With Antineutrophil Cytoplasmic Antibody-Associated Vasculitis: A 15-Year Multicenter Retrospective Study.

Background: Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is a multisystem autoimmune disease with small-vessel involvement. In AAV, microscopic polyangiitis (MPA) and granulomatosis with polyangiitis (GPA) are major clinicopathologic variants. In addition, myeloperoxidase (MPO) and proteinase 3 (PR3) are major target antigens. The objective of the study was to explore the predictive factors for long-term survival in AAV patients. Materials and Methods: A multicenter retrospective study was carried out on 407 patients between 2005 and 2020. Clinical parameters were obtained from laboratory tests including the ANCA types, antinuclear antibody (ANA), extractable nuclear antigen (ENA), anti-streptolysin O (ASO), glomerular filtration rate (GFR), and the laboratory examinations for the blood routine, liver function, renal function, and immunity, etc. The data for clinical parameters were collected from electronic medical records (EMRs), and the data for patient survival were acquired through regular follow-up. The association of clinical parameters with overall survival (OS) along with 3-year and 5-year survival rates was analyzed, and the nomogram as a predictive model was established according to the analysis results. Results: In the present study, 336 (82.6%) patients and 46 (11.3%) patients were diagnosed with MPA and GPA, respectively. The mean and median OS for all the patients were 2,285 and 2,290 days, respectively. The 1-year, 3-year, 5-year, and 10-year cumulative survival rates for all the patients were 84.2%, 76.3%, 57.2%, and 32.4%, respectively. Univariate and multivariate survival analyses indicated that the independent prognostic factors included age, pathological categories (MPA, GPA, and other types), serum ANCA types (negative or positive for MPO and/or PR3), ANA, ASO, GFR, lymphocyte, neutrophil-to-lymphocyte ratio (NLR), and C-reactive protein (CRP), and these clinical parameters except for ASO were used to construct a nomogram. The nomogram for 3-year and 5-year survival rates had a C-index of 0.721 (95% CI 0.676-0.766). The calibration curves showed that the predicted values of the nomogram for 3-year and 5-year survival rates were generally consistent with practical observed values, and decision curve analysis (DCA) further demonstrated the practicability and accuracy of the predictive model. Conclusion: Laboratory tests at diagnosis have great significance in the prediction of long-term survival in AAV patients.

[Robot-assisted laparoscopic radical prostatectomy helps improve postoperative urinary continence recovery in patients with prostate cancer].

OBJECTIVE: To observe the recovery of urinary continence through postoperative rehabilitation training after robot-assisted laparoscopic radical prostatectomy (RARP) versus that after traditional laparoscopic radical prostatectomy (LRP). METHODS: This study included 64 cases of urinary incontinence after surgically treated for PCa from May 2017 to February 2021, 32 by RARP and the other 32 by LRP as the controls. All the patients received standard urinary continence rehabilitation training and routine nursing care postoperatively, followed by comparison of the rate and time of urinary incontinence recovery and the patients' scores on the quality of life (QOL) and satisfaction with treatment between the two groups. RESULTS: There were no statistically significant differences in age, PSA level or pathological stage between the two groups of patients (P > 0.05). After standard urinary continence rehabilitation training, the patients in the RARP group, compared with those in the LRP control, showed a lower grade of urinary incontinence (χ2 = 6.483, P = 0.039), a shorter mean duration of urinary incontinence per day, an earlier recovery of urinary continence (χ2 = 4.73, P = 0.030 at 1－3 months; χ2 = 12.696, P < 0.001 at 4－6 months), a higher rate of overall recovery (χ2 = 13.396, P = 0.004), and higher scores on QOL and satisfaction with treatment. CONCLUSION: RARP can effectively improve the recovery from postoperative urinary incontinence in PCa patients.

Metabolome and proteome analyses reveal transcriptional misregulation in glycolysis of engineered E. coli.

Synthetic metabolic pathways are a burden for engineered bacteria, but the underlying mechanisms often remain elusive. Here we show that the misregulated activity of the transcription factor Cra is responsible for the growth burden of glycerol overproducing E. coli. Glycerol production decreases the concentration of fructose-1,6-bisphoshate (FBP), which then activates Cra resulting in the downregulation of glycolytic enzymes and upregulation of gluconeogenesis enzymes. Because cells grow on glucose, the improper activation of gluconeogenesis and the concomitant inhibition of glycolysis likely impairs growth at higher induction of the glycerol pathway. We solve this misregulation by engineering a Cra-binding site in the promoter controlling the expression of the rate limiting enzyme of the glycerol pathway to maintain FBP levels sufficiently high. We show the broad applicability of this approach by engineering Cra-dependent regulation into a set of constitutive and inducible promoters, and use one of them to overproduce carotenoids in E. coli.

The Landscape of Cell-Free HBV Integrations and Mutations in Cirrhosis and Hepatocellular Carcinoma Patients.

PURPOSE: Intratumoral hepatitis B virus (HBV) integrations and mutations are related to hepatocellular carcinoma (HCC) progression. Circulating cell-free DNA (cfDNA) has shown itself as a powerful noninvasive biomarker for cancer. However, the HBV integration and mutation landscape on cfDNA remains unclear. EXPERIMENTAL DESIGN: A cSMART (Circulating Single-Molecule Amplification and Resequencing Technology)-based method (SIM) was developed to simultaneously investigate HBV integration and mutation landscapes on cfDNA with HBV-specific primers covering the whole HBV genome. Patients with HCC (n = 481) and liver cirrhosis (LC; n = 517) were recruited in the study. RESULTS: A total of 6,861 integration breakpoints including TERT and KMT2B were discovered in HCC cfDNA, more than in LC. The concentration of circulating tumor DNA (ctDNA) was positively correlated with the detection rate of these integration hotspots and total HBV integration events in cfDNA. To track the origin of HBV integrations in cfDNA, whole-genome sequencing (WGS) was performed on their paired tumor tissues. The paired comparison of WGS data from tumor tissues and SIM data from cfDNA confirmed most recurrent integration events in cfDNA originated from tumor tissue. The mutational landscape across the whole HBV genome was first generated for both HBV genotype C and B. A region from nt1100 to nt1500 containing multiple HCC risk mutation sites (OR > 1) was identified as a potential HCC-related mutational hot zone. CONCLUSIONS: Our study provides an in-depth delineation of HBV integration/mutation landscapes at cfDNA level and did a comparative analysis with their paired tissues. These findings shed light on the possibilities of noninvasive detection of virus insertion/mutation.

Multi-omics Analysis of CRISPRi-Knockdowns Identifies Mechanisms that Buffer Decreases of Enzymes in E. coli Metabolism.

Enzymes maintain metabolism, and their concentration affects cellular fitness: high enzyme levels are costly, and low enzyme levels can limit metabolic flux. Here, we used CRISPR interference (CRISPRi) to study the consequences of decreasing E. coli enzymes below wild-type levels. A pooled CRISPRi screen with 7,177 strains demonstrates that metabolism buffers fitness defects for hours after the induction of CRISPRi. We characterized the metabolome and proteome responses in 30 CRISPRi strains and elucidated three gene-specific buffering mechanisms: ornithine buffered the knockdown of carbamoyl phosphate synthetase (CarAB) by increasing CarAB activity, S-adenosylmethionine buffered the knockdown of homocysteine transmethylase (MetE) by de-repressing expression of the methionine pathway, and 6-phosphogluconate buffered the knockdown of 6-phosphogluconate dehydrogenase (Gnd) by activating a bypass. In total, this work demonstrates that CRISPRi screens can reveal global sources of metabolic robustness and identify local regulatory mechanisms that buffer decreases of specific enzymes. A record of this paper's transparent peer review process is included in the Supplemental Information.

[Distribution Characteristics and Source Identification of Organophosphate Esters in Key Waters Areas of Taihu Lake].

Organophosphate esters (OPEs) are ubiquitous in the environment and pose potential risks to ecosystems in that they produce cytotoxicity, genetic toxicity, developmental toxicity, neurotoxicity, and the endocrine disrupting effect. This study aimed to investigate the distribution, sources, and ecological risk of thirteen OPEs in industrial wastewater, influents and effluents of wastewater treatment plants, and the surface waters of key areas (Changzhou City) in Taihu Lake. The results showed that ∑OPEs in industrial wastewater ranged from 91.70-840.52 ng·L-1. The profiles varied from different industries; however, tris (1-chloro-2-propyl) phosphate (TCPP), triethyl phosphate (TEP), and tris (2-chloroethyl) phosphate (TCEP) were the dominant compounds. The ∑OPEs from the wastewater treatment plants were relatively higher, ranging from 1859.59-2778.57 ng·L-1. They are rather resistant to traditional wastewater treatment techniques. The removal rate of ∑OPEs, aryl OPEs, and chlorinated OPEs was 14.97%, 97.91%, and 4.37% (for W1) and 17.32%, 90.83%, and 5.40% (for W2), respectively. The concentration of OPEs in the surface water was in the range of 219.47-689.85 ng·L-1, which was lower than that in wastewater. In addition, TEP, TCPP and TCEP predominated in the surface water samples. A principle component analysis (PCA) indicated that the industrial wastewater, emissions of vehicles, and the release of building materials may be the main sources of OPEs in the study area. A risk quotient was used to assess the ecological risk of OPEs to aquatic organisms in the surface water. Most of the OPEs showed low ecological risk, but 2-ethylhexyl diphenyl phosphate (EHDPP) posed a moderate risk in some sites, and the possible ecological effects caused by the OPEs should not be ignored in the key areas of Taihu Lake.

[Pollution Characteristics and Risk Assessment of Typical Organophosphate Esters in Beijing Municipal Wastewater Treatment Plant and the Receiving Water].

Organophosphate esters (OPEs) are ubiquitous in the environment and pose a potential threat to ecosystems and human health. A method for the determination of eight OPEs by ultra-performance liquid chromatography-tandem mass spectrometer (UPLC-MS/MS) was established. The recovery rates of eight target compounds with different solid-phase extraction columns, different eluents, and different eluent volumes were compared. The results showed that using ENVI-18 column enrichment, OPEs were eluted with 8 mL acetonitrile containing 25% (volume fraction) dichloromethane, and the labeled recovery rate of the target compound was 92.5%-102.2%. The recoveries of different matrix samples were 88.5%-116.1% and relative standard deviation was 1.7%-9.9%. The concentration range of 8 different detectable organophosphate esters in the effluent of sewage treatment plant is 85.9-235.4 ng·L-1 during the six-day sampling process, permissive river downstream of the six-day ΣOPEs average total concentration was 130.3 ng·L-1, higher than the 119.4 ng·L-1 upstream water concentration, but lower than the sewage treatment plant effluent concentration of total 162.5ng·L-1. The study shows that the sewage treatment plant cannot completely remove OPEs; for triethyl phosphate (TEP) and 3 (2-ethyl hexyl) phosphate ester (TEHP) there exists a negative removal phenomenon, whereas for other OPEs the removal rate was between 14.1% and 84.9%, and the total ΣOPEs removal rate by the sewage plant was 50.0%. The TPhP in the effluent of the sewage treatment plant has medium environmental risk (RQ>0.1), and other organophosphates have low environmental risk (RQ<0.1); however, the long-term mixing effects of organophosphate esters on the ecosystem of the receiving river should not be ignored.

miR-143 inhibits renal cell carcinoma cells metastatic potential by suppressing ABL2.

Although micro RNA (miRNA) expression profiles are widely investigated in renal cell carcinoma (RCC), their potential roles for affecting RCC initiation and progression remain largely unknown. Here, we examined the aberrant expression profiles of miRNAs inhuman metastatic RCC tissues based on Gene Expression Omnibus (GSE37989). We further validated them iRNAs expression data in the largest clinical dataset: The Cancer Genome Atlas (TCGA). And cell adhesion and migration abilities and epithelial me senchymal transition (EMT) related proteins were assessed in both normal and tumor RCC cell lines. We suggest that hsa-miR-143 is a potential tumor suppressor in RCC as its down regulation positively correlated with adverse prognosis. Biologically, cell adhesion, migration, and EMT were dramatically inhibited by miR-143. Mechanistically, we found that miR-143 targets ABL proto-oncogene 2 (ABL2), which was also found to be an indicator for poor survival in TCGA database. Our results have important implications in understanding functions of miRNAs in metastatic RCC and will provide a basis for further clinical application.

Evaluation of 18F-DCFPyL PSMA PET/CT for Prostate Cancer: A Meta-Analysis.

BACKGROUND: To systematically review the clinical value of 18F-DCFPyL prostate-specific membrane antigen positron emission tomography/computed tomography (PSMA PET/CT) in the diagnosis of prostate cancer (PCa). METHODS: Literature concerning 18F-DCFPyL PSMA PET/CT in the diagnosis of prostate cancer published from 2015 to 2020 was electronically searched in the databases including PubMed and Embase. Statistical analysis was carried out with STATA 15 software, and the quality of included studies was tested with quality assessment of diagnostic accuracy studies (QUADAS) items. The heterogeneity of the included data was tested. RESULTS: In total, nine pieces of literature involving 426 patients met the inclusion criteria. The heterogeneity of the study group was not obvious. The SEN, SPE, LR+, LR-, DOR as well as AUC of 18F-DCFPyL PSMA PET/CT diagnosis of prostate cancer were 0.91, 0.90, 8.9, 0.10, 93, and 0.93. The pooled DR of 18F-DCFPyL labeled PSMA PET/CT in PCa was 92%. The pooled DR was 89% for PSA≥0.5 ng/ml and 49% for PSA < 0.5ng/ml. CONCLUSION: 18F-DCFPyL PSMA PET/CT had good sensitivity and specificity for the diagnosis of prostate cancer. The DR of 18F-DCFPyL PSMA PET/CT was correlated with PSA value. Further large-sample, high-quality studies were needed.

Lewis Acid Catalyzed [4 + 2] Cycloaddition of N-Tosylhydrazones with ortho-Quinone Methides.

A formal [4 + 2] cycloaddition of N-tosylhydrazones with ortho-quinone methides was developed, affording the facile synthesis of diverse 1,3-oxazine derivatives under mild conditions. In this transformation, N-tosylhydrazones are used as a 1,2-dipole synthon under base-free conditions. Moreover, the substrate scope is broad, and the products are formed with high diastereoselectivities in most of the cases.

Application of self-care based on full-course individualized health education in patients with chronic heart failure and its influencing factors.

BACKGROUND: The treatment of heart failure not only needs to relieve the clinical symptoms and improve the quality of life for patients but also needs to select scientific and reasonable ways to prevent or delay the progression of the disease, thus reducing the mortality and hospitalization rate. Although the previous regimen can effectively relieve symptoms in the early stage of treatment, long-term use may cause adverse events, such as arrhythmia, and even increase mortality. Therefore, conventional treatment cannot meet the actual health needs of patients, and scientific nursing intervention is very necessary. AIM: To investigate the application of self-care based on full-course individualized health education (FCIHE) and its influencing factors in patients with chronic heart failure (CHF). METHODS: We enrolled CHF patients who were admitted to our center between September 2015 and June 2016 and divided them into an intervention group (n = 50) and control group (n = 50) using a random number table. Routine nursing care was applied to the control group, and FCIHE was offered to the intervention group. The self-care behavior, 6-min walking distance (6MWD), and 36-item short form health survey (SF-36) scores were compared between the two groups. The influencing factors of the self-care were also analyzed. RESULTS: The 6MWD was not significantly different between the two groups at admission (P > 0.05); however, at 3 and 6 mo after discharge, 6MWD was significantly increased, and it was significantly longer in the intervention group (P < 0.05). The scores for self-care behavior showed no significant difference at admission between the two groups (P > 0.05); however, at 3 and 6 mo after discharge, the total scores for self-care maintenance, management, confidence, and behavior of the intervention group were significantly higher than those of the control group (P < 0.05). There were no significant differences in the SF-36 scores at admission (P > 0.05); however, at 3 mo and 6 mo after discharge, the scores for all eight subscales, including physical functioning, role limitations due to physical problems, bodily pain, general health perceptions, vitality, social functioning, role-limitations due to emotional problems, and mental health, were significantly higher in the intervention group (P < 0.05). As shown by logistic regression analysis, the influencing factors of self-care mainly included age, cardiac function class, and education background (odds ratio > 1; all P < 0.05). CONCLUSION: FCIHE improved self-care behavior and cardiac function in CHF patients. Age, cardiac function, and education level affected the implementation of self-care among CHF patients.

CRISPRi-Based Downregulation of Transcriptional Feedback Improves Growth and Metabolism of Arginine Overproducing E. coli.

Removing transcriptional feedback regulation of metabolic pathways is a classical approach to enhance overproduction of chemicals in microbes. However, disrupting transcriptional regulation can have broad physiological consequences that decrease cellular growth and productivity. Here, we compared downregulation and deletion of the transcriptional repressor ArgR in arginine overproducing Escherichia coli. Different levels of ArgR downregulation were achieved with CRISPR interference (CRISPRi) and resulted in 2-times higher growth rates compared to deletion of ArgR, while specific arginine production was similar (∼2 mmol gDW-1 h-1). Metabolomics and proteomics data revealed that poor growth of the ArgR deletion strain was caused by a limitation of pyrimidine nucleotide biosynthesis, because a 17-fold overexpression of ornithine carbamoyltransferase (ArgI) perturbed the arginine-pyrimidine branch point. These results demonstrate that overexpression of enzymes in an engineered pathway can impair metabolism of the host, especially in the case of branch-point enzymes. Thus, balancing enzyme levels is important to optimize industrial microbes, and CRISPRi of a transcription factor is a versatile tool for this purpose.

Boron-Catalyzed Azide Insertion of α-Aryl α-Diazoesters.

A challenging metal-free azide insertion of α-aryl α-diazoesters in the presence of B(C6F5)3 (5 mol %) was developed for the first time. The reaction features an easy operation, wide substrate scope, and mild conditions and affords the corresponding products in moderate to high yields. More importantly, alkene and alkyne functional groups were well tolerated because no cyclopropanation or cyclopropenation was observed. Furthermore, the corresponding azide products could be converted to primary amines or 1,2,3-triazole derivatives after simple transformations.

[Simultaneous determination of 8 constituents of Desmodii Styracifolii Herba in rat bile with HPLC-MS/MS and their excretion pharmacokinetics].

To establish a HPLC-MS/MS quantitative method for analysis of 8 constituents in rat bile. The method was applied in the biliary excretion study after oral administration of Desmodii Styracifolii Herba extract. The rat bile samples were collected during 0-1, 1-2, 2-4, 4-6, 6-8, 8-12, 12-24 h. Diamonsil C18column (4.6 mm×150 mm, 5 µm) was adopted and eluted with methanol and 0.01% acetic acid in a gradient mode. The flow rate was 0.8 mL•min⁻¹, and the column temperature was set at 40 ℃. The detection was carried out by a triple quadrupole linearion trap mass spectrometer in the negative ion mode with an electrospray source. Multiple reactions monitoring (MRM) mode was employed. The calibration curves showed a good linearity, with correlation coefficients (r≥0.991 5) for all of the analytes within the concentration range. The intra-day and inter-day precisions (RSD) were both less than 15%, and the accuracies (RE) ranged from -15% to 15%. The method had a good stability, and was suitable for the content determination of 8 constituents in rat bile. The extraction recoveries of the 8 analytes were more than 63.2%, and the RSD of IS-normalized matrix factors were no more than 15%, which met the requirements for analysis. According to the biliary excretion study, the bile excretion rates of the 8 analytes were relatively low, with great difference among the individuals. The results showed that the established method was simple, selective and specific, thus could be applied in the biliary excretion study of the 8 analytes in rat bile.

A long-term survivor with esophageal melanoma and pulmonary metastasis after single-stage esophagectomy and lobectomy: Case report and literature review.

RATIONALE: The optimal therapeutic regimen for primary malignant melanoma of the esophagus (PMME) need to be further elucidated. Besides, the efficacy of surgery for PMME with remote metastasis is uncertain for its rarity. PATIENT CONCERNS: Herein a previously healthy patient was admitted for dysphagia and fatigue, without significant weight loss. DIAGNOSES: The pathological and molecular tests revealed his diagnosis of BRAF-mutant, advanced PMME with localized pulmonary metastasis. INTERVENTIONS: Single-stage Ivor-Lewis esophagectomy and lobectomy were performed successfully, followed by 4 cycles of conventional chemotherapy, and concurrent high-dose interferon lasting for 1 year. OUTCOMES: The patient survived without logo-regional recurrence or remote metastasis during the follow up of two and a half years up to now. LESSONS: Timely resection of localized primary and metastatic lesions might deliver a chance to obtain better prognosis for selected PMME patients; however, high-quality trials with longer follow-up are needed.