BPL3 binds the long non-coding RNA nalncFL7 to suppress FORKED-LIKE7 and modulate HAI1-mediated MPK3/6 dephosphorylation in plant immunity.

RNA-binding proteins (RBPs) participate in a diverse set of biological processes in plants, but their functions and underlying mechanisms in plant-pathogen interactions are largely unknown. We previously showed that Arabidopsis thaliana BPA1-LIKE PROTEIN3 (BPL3) belongs to a conserved plant RBP family and negatively regulates reactive oxygen species (ROS) accumulation and cell death under biotic stress. In this study, we demonstrate that BPL3 suppresses FORKED-LIKE7 (FL7) transcript accumulation and raises levels of the cis-natural antisense long non-coding RNA (lncRNA) of FL7 (nalncFL7). FL7 positively regulated plant immunity to Phytophthora capsici while nalncFL7 negatively regulated resistance. We also showed that BPL3 directly binds to and stabilizes nalncFL7. Moreover, nalncFL7 suppressed accumulation of FL7 transcripts. Furthermore, FL7 interacted with HIGHLY ABA-INDUCED PP2C1 (HAI1), a type 2C protein phosphatase, and inhibited HAI1 phosphatase activity. By suppressing HAI1 activity, FL7 increased the phosphorylation levels of MITOGEN-ACTIVATED PROTEIN KINASE 3 (MPK3) and MPK6, thus enhancing immunity responses. BPL3 and FL7 are conserved in all plant species tested, but the BPL3-nalncFL7-FL7 cascade was specific to the Brassicaceae. Thus, we identified a conserved BPL3-nalncFL7-FL7 cascade that coordinates plant immunity.

Real-time fluorescence imaging with indocyanine green during laparoscopic duodenum-preserving pancreatic head resection.

BACKGROUND: Laparoscopic duodenum-preserving pancreatic head resection (LDPPHR) is a surgical method used to treat benign and low-grade malignant pancreatic head tumors. This study aimed to determine the protective effect of common bile duct in LDPPHR using indocyanine green (ICG) fluorescence imaging. METHODS: A retrospective analysis of 30 patients treated with LDPPHR at the Second Affiliated Hospital of Nanchang University between January 2015 and November 2022 was performed. Patients were divided into two groups based on ICG use: ICG and non-ICG. RESULTS: Thirty patients received LDPPHR, 11 males and 19 females, and the age was 50.50 (M (IQR)) years (range: 19-76 years). LDPPHR was successfully performed in 27 (90 %) patients, LPD was performed in 1 (3 %) patient, and laparotomy conversion was performed in 2 (7 %) patients. One patient (3 %) died 21 days after surgery. The incidence of intraoperative bile duct injury in the ICG group was lower than that in the non-ICG group (10 % vs 60 %, P = 0.009), and the operation time in the ICG group was shorter than that in the non-ICG group (311.9 ± 14.97 vs 338.05 ± 18.75 min, P < 0.05). Postoperative pancreatic fistula occurred in 16 patients (53 %), including 10 with biochemical leakage (62.5 %), four with grade B (25 %), and two with grade C (12.5 %). Postoperative bile leakage occurred in four patients (13 %). CONCLUSIONS: The ICG fluorescence imaging technology in LDPPHR helps protect the integrity of the common bile duct and reduce the occurrence of intraoperative bile duct injury, postoperative bile leakage, and bile duct stenosis.

Hierarchically engineered nanostructures from compositionally anisotropic molecular building blocks.

The inability to synthesize hierarchical structures with independently tailored nanoscale and mesoscale features limits the discovery of next-generation multifunctional materials. Here we present a predictable molecular self-assembly strategy to craft nanostructured materials with a variety of phase-in-phase hierarchical morphologies. The compositionally anisotropic building blocks employed in the assembly process are formed by multicomponent graft block copolymers containing sequence-defined side chains. The judicious design of various structural parameters in the graft block copolymers enables broadly tunable compositions, morphologies and lattice parameters across the nanoscale and mesoscale in the assembled structures. Our strategy introduces advanced design principles for the efficient creation of complex hierarchical structures and provides a facile synthetic platform to access nanomaterials with multiple precisely integrated functionalities.

A Phytophthora sojae CRN effector mediates phosphorylation and degradation of plant aquaporin proteins to suppress host immune signaling.

Phytophthora genomes encode a myriad of Crinkler (CRN) effectors, some of which contain putative kinase domains. Little is known about the host targets of these kinase-domain-containing CRNs and their infection-promoting mechanisms. Here, we report the host target and functional mechanism of a conserved kinase CRN effector named CRN78 in a notorious oomycete pathogen, Phytophthora sojae. CRN78 promotes Phytophthora capsici infection in Nicotiana benthamiana and enhances P. sojae virulence on the host plant Glycine max by inhibiting plant H2O2 accumulation and immunity-related gene expression. Further investigation reveals that CRN78 interacts with PIP2-family aquaporin proteins including NbPIP2;2 from N. benthamiana and GmPIP2-13 from soybean on the plant plasma membrane, and membrane localization is necessary for virulence of CRN78. Next, CRN78 promotes phosphorylation of NbPIP2;2 or GmPIP2-13 using its kinase domain in vivo, leading to their subsequent protein degradation in a 26S-dependent pathway. Our data also demonstrates that NbPIP2;2 acts as a H2O2 transporter to positively regulate plant immunity and reactive oxygen species (ROS) accumulation. Phylogenetic analysis suggests that the phosphorylation sites of PIP2 proteins and the kinase domains of CRN78 homologs are highly conserved among higher plants and oomycete pathogens, respectively. Therefore, this study elucidates a conserved and novel pathway used by effector proteins to inhibit host cellular defenses by targeting and hijacking phosphorylation of plant aquaporin proteins.

The Synergistic Effects between Liquid Crystal and Crystalline Phase on Photo-Responsive Elastomers toward Quick Photo-Responsive Performance.

Adopting only a small amount of azobenzene molecular to design liquid crystal photo-responsive materials capable of quick response and flexible adjustability is in high demand but is challenging. Herein, azobenzenemolecules into polyurethane elastomer containing crystalline structure for preparing azobenzene liquid-crystal elastomers (ALCEs) are demonstrated and this phenomenon of the synergistic effects between liquid crystal and crystalline phase is discovered. The key point of the work is that the synthetic ALCEs can utilize the reversible isomerism capability of azobenzene molecules under light irradiation, which can pry the motion of the macromolecular crystalline region in system to realize the large macroscopic deformation of the photo-responsive behavior. Obviously, the ALCEs sample containing azobenzene molecule and polyethylene glycol crystallization can quickly bend, illuminated by ultraviolet light and rapidly straighten under green light. Under the same ultraviolet irradiation, the bending speed, final bending angle, recovery rate and recovery ratio of ALCEs are larger than that of ALCEs without any crystalline structure. This ALCEs based on the synergistic effects between liquid crystal and crystalline phase can break through the current dilemma that the application of traditional azobenzene photo-responsive materials is limited by their concentration, greatly expanding the design thought and their scope of application.

Laparoscopic anatomical left hemihepatectomy guided by middle hepatic vein in the treatment of left hepatolithiasis with a history of upper abdominal surgery.

BACKGROUND: This study aimed to investigate the safety and efficacy of laparoscopic anatomical left hemihepatectomy guided by the middle hepatic vein (MHV) for the treatment of patients with hepatolithiasis who had a history of upper abdominal surgery. METHODS: Retrospective data analysis was performed on patients who underwent laparoscopic left hepatectomy for hepatolithiasis and with previous upper abdominal surgery at the Second Affiliated Hospital of Nanchang University from January 2018 to April 2022. According to the different surgical approaches, patients were divided into laparoscopic anatomical left hepatectomy guided by the MHV group (MHV-AH group) and laparoscopic traditional anatomical left hepatectomy not guided by the MHV group (non-MHV-AH group). RESULTS: This study included 81 patients, with 37 and 44 patients in the MHV-AH and non-MHV-AH groups, respectively. There was no significant difference in the basic information between the two groups. Five cases were converted to laparotomy, and the remaining were successfully completed under laparoscopy. Compared to the non-MHV-AH group, the MHV-AH group had a slightly longer operation time (319.30 min vs 273.93 min, P = 0.032), lower bile leakage rate (5.4% vs 20.5%, P = 0.047), stone residual rate (2.7% vs 20.5%, P = 0.015), stone recurrence rate (5.4% vs 22.7%, P = 0.028), and cholangitis recurrence rate (2.7% vs 22.7%, P = 0.008).There were no significant differences in the results of other observation indices between the groups. CONCLUSIONS: Laparoscopic anatomical left hepatectomy guided by the MHV is safe and effective in the treatment of left hepatolithiasis with a history of upper abdominal surgery. It does not increase intraoperative bleeding and reduces the risk of postoperative bile leakage, residual stones, stone recurrence, and cholangitis recurrence.

The microRNAs in the antennae of Apolygus lucorum (Hemiptera: Miridae): Expression properties and targets prediction.

MicroRNAs (miRNAs) regulate gene expression and contribute to numerous physiological processes. However, little is known about the functions of miRNAs in insect chemosensation. In this study, nine small RNA libraries were constructed and sequenced from the antennae of nymphs, adult males, and adult females of Apolygus lucorum. In total, 399 (275 known and 124 novel) miRNAs were identified. miR-7-5p_1 was the most abundant miRNA. Altogether, 69,708 target genes related to biogenesis, membrane, and binding activities were predicted. In particular, 15 miRNAs targeted 16 olfactory genes. Comparing the antennae of nymphs and adult males and females, 94 miRNAs were differentially expressed. Alternatively, a subset of differentially expressed miRNAs was verified by qPCR, supporting the reliability of the sequencing results. This study provides a global miRNA transcriptome for the antennae of A. lucorum and valuable information for further investigations of the functions of miRNAs in the regulation of chemosensation.

Phytophthora infection signals-induced translocation of NAC089 is required for endoplasmic reticulum stress response-mediated plant immunity.

Plants deploy various immune receptors to recognize pathogen-derived extracellular signals and subsequently activate the downstream defense response. Recently, increasing evidence indicates that the endoplasmic reticulum (ER) plays a part in the plant defense response, known as ER stress-mediated immunity (ERSI), that halts pathogen infection. However, the mechanism for the ER stress response to signals of pathogen infection remains unclear. Here, we characterized the ER stress response regulator NAC089, which was previously reported to positively regulate programed cell death (PCD), functioning as an ERSI regulator. NAC089 translocated from the ER to the nucleus via the Golgi in response to Phytophthora capsici culture filtrate (CF), which is a mixture of pathogen-associated molecular patterns (PAMPs). Plasma membrane localized co-receptor BRASSINOSTEROID INSENSITIVE 1-associated receptor kinase 1 (BAK1) was required for the CF-mediated translocation of NAC089. The nuclear localization of NAC089, determined by the NAC domain, was essential for immune activation and PCD. Furthermore, NAC089 positively contributed to host resistance against the oomycete pathogen P. capsici and the bacteria pathogen Pseudomonas syringae pv. tomato (Pst) DC3000. We also proved that NAC089-mediated immunity is conserved in Nicotiana benthamiana. Together, we found that PAMP signaling induces the activation of ER stress in plants, and that NAC089 is required for ERSI and plant resistance against pathogens.

Seeded Growth of Gold-Copper Janus Nanostructures as a Tandem Catalyst for Efficient Electroreduction of CO2 to C2+ Products.

Gold-copper (Au-Cu) Janus nanostructures (Au-Cu Janus NSs) are successfully prepared using N-oleyl-1,3-propanediamine as capping agent and Cu(acac)2 as the precursor in a typical seeded growth strategy. By preferably depositing Cu atoms on one side of concave cubic Au seeds, the Cu part gradually grows larger as more Cu precursors are added, making the size tuning feasible in the range of 74-156 nm. When employed as an electrocatalyst for electrochemical CO2 reduction (CO2 RR), the Au-Cu Janus NSs display superior performance to Au@Cu core-shell NSs and Cu NPs in terms of C2+ products selectivity (67%) and C2+ partial current density (-0.29 A cm-2 ). Combined experimental verification and theoretical simulations reveal that CO spillover from Au sites to the nearby Cu counterparts would enhance CO coverage and thus promote C-C coupling, highlighting the unique structural advantages of the Au-Cu Janus NSs toward deep reduction of CO2 . The current work provides a facile strategy to fabricate tandem catalyst with a Janus structure and validates its structural advantages toward CO2 RR, which are of critical importance for the rational design of efficient CO2 RR catalyst.

In Situ Crumpling of Gold Nanosheets into Spherical Three-Dimensional Architecture: Probing the Aggregation-Induced Enhancement in Photothermal Properties.

Assembling two-dimensional noble metal nanocrystals into a three-dimensional mesoporous structure is of great value to solve the re-stacking issue for the practical application, which still remains a challenging technique. Herein, we report the one-pot fabrication of gold (Au) nanostructures with a crumpled paper ball-like morphology (Au NCPBs). The success of current work relies on the use of glutathione to crumple the branched Au nanosheets formed during the early stage, into spherical three-dimensional architecture, where the nanosheets are assembled with a mesoporous structure without intimate contact. When working as the agent toward photothermal conversion, the Au NCPBs exhibit enhanced photothermal conversion efficiency (η = 19.9%), as compared to that of flat and wrinkled Au nanosheets. Such an enhancement should be owing to the aggregation-induced effect, where the shortened inter-sheet distance contributes to an increased coupling between the plasmon oscillations/fields of the interacting Au nanosheets. The present study offers a feasible strategy to create spherical architecture of crumpled Au nanosheets and validates their structural advantage in photothermal applications, which could be potentially extended to other metals or alloys.

Psychological distress of cancer patients caused by treatment delay during the COVID-19 pandemic in China: A cross-sectional study.

OBJECTIVE: The currents study sought to explore the impact of treatment delay on the mental health for patients with cancer during the 2019 coronavirus disease (COVID-19) pandemic. METHODS: Travel restrictions were imposed in most areas of the country between 23 January 2020 and 25 February 2020 owing to the COVID-19 epidemic. Travel restrictions were lifted from 26 February 2020 to 12 March 2020. The number of new confirmed cases significantly reduced after 12 March 2020. Study participants, comprised of individuals from three distinct groups: (1) 835 cancer patients who attended Zhejiang Cancer Hospital between 26 February 2020 and 12 March 2020; (2) 185 healthy volunteers recruited between 26 February 2020 and 12 March 2020; (3) 168 cancer patients who attended the hospital during the non-epidemic period (after 12 March 2020). Two outcome measures including patients' posttraumatic stress responses and general psychological distress (GPD) were assessed using the Chinese versions of the Impact of Events Scale-Revised and the Kessler Psychological Distress Scale (K10). Treatment delay was assessed via counting the time interval from diagnosis to treatment initiation, or from planned treatment date to actual date of therapy. Communication satisfaction was evaluated via a self-report questionnaire. An independent sample t-test or Wilcoxon rank sum test was used for comparison. Statistical analysis included Chi-square test, Mann-Whitney test and multivariate logistic regression. RESULTS: All 1188 participants (835 patients with cancer and 185 controls during the outbreak, and 168 patients with cancer during the non-epidemic period) completed and submitted the questionnaires. A positive association was observed between treatment delays and increased GPD levels (OR 1.716; 95% confidence interval ,CI 1.254-2.348; p = 0.001) as well as posttraumatic stress disorder (PTSD) symptoms (OR: 1.545, 95% CI: (1.166-2.047), p = 0.002). Patients who reported good communication with their doctors showed a significantly lower risk of GPD (OR: 0.526, 95% CI (0.348-0.794), p = 0.002) and PTSD (OR: 0.683, 95% CI (0.490-0.951), p = 0.024) compared with patients who reported unsatisfactory communication or had no contact with their doctors. Multivariate logistic regression analysis showed that treatment at a local hospital, treatment delays and unsatisfactory or no communication with cancer-care professionals were significantly correlated with severe GPD and PTSD symptoms of patients (all p ≤ 0.05). CONCLUSION: The findings indicate that cancer patients who underwent treatment delays during the COVID-19 pandemic may become vulnerable to psychological distress. The results showed that effective communication with doctors and cancer-care professionals during outbreak significantly reduces GPD levels and PTSD symptoms.

Oxime-Urethane Structure-Based Dynamically Crosslinked Polyurethane with Robust Reprocessing Properties.

Crosslinked polyurethane with excellent mechanical property, solvent resistance, and transparency has become one of the most widely used materials. However, the presence of chemical crosslinks makes it difficult to be reprocessed once molded, which largely restricts its recycling and reusing, resulting in the serious waste problems. Therefore, it is of great significance to prepare a new type of crosslinked polyurethane with reprocessing function. In this work, a novel reprocessable polyurethane (DOPU) based on reversible dibutanone oxime-carbamate bonds is facilely prepared. The gel fraction of DOPUs is all higher than 95%, endowing it with excellent solvent resistance. Meanwhile, the visible light transmittance of DOPUs can reach up to 97.48%. After four thermal recycles, the tensile strength and elongation at break of recycled DOPUs can still remain at 3.21 MPa and 219.09%, respectively. Importantly, the synthesized DOPUs exhibit excellent elastic shape memory and permanent shape reconstruction properties under thermal stimulation. The dibutanone oxime-carbamate bonds can also be degraded under UV irradiation, making this material easily degradable. Hence, this material has potential applications in coatings, elastomers, and some other fields.

EPS8 supports pancreatic cancer growth by inhibiting BMI1 mediated proteasomal degradation of ALDH7A1.

Aldehyde dehydrogenase 7 family member A1 (ALDH7A1) is an enzyme catalyzing lipid peroxidation of fatty aldehydes. It plays a critical role in sustaining high oxygen consumption rate (OCR) and ATP production in pancreatic ductal adenocarcinoma (PADC). However, why PADC cells maintain a relatively high level of ALDH7A1 concentration is still not well understood. In the current study, we explored the interplay between epidermal growth factor receptor kinase substrate 8 (EPS8) and ALDH7A1 in PADC cells. PADC cell lines MIA PaCa-2 and AsPANC-1 were used for in vitro and in vivo studies. The co-IP assay showed mutual interactions between Flag-EPS8 and Myc-ALDH7A1 in both MIA PaCa-2 and AsPANC-1 cells. EPS8 knockdown resulted in decreased ALDH7A1 protein levels and increased poly-ubiquitination. An interaction was observed between ALDH7A1 and BMI1 but not between BMI1 and EPS8. BMI1 knockdown reduced ALDH7A1 poly-ubiquitination and degradation caused by EPS8 knockdown. Dual EPS8 and ALDH7A1 knockdown had a synergistic effect on suppressing PADC cell proliferation in vitro and in vivo. In conclusion, this study revealed that EPS8 supports PADC growth by interacting with ALDH7A1 and inhibiting BMI1 mediated proteasomal degradation of ALDH7A1.

Rapid Access to Diverse Multicomponent Hierarchical Nanostructures from Mixed-Graft Block Copolymers.

Multicomponent nanostructured materials assembled from molecular building blocks received wide attention due to their precisely integrated multifunctionalities. However, discovery of these materials with desirable composition and morphology was limited by their low synthetic scalability and narrow structural tuning window with given building blocks. Here, we report a scalable and diversity-oriented synthetic approach to hierarchically structured nanomaterials based on a few readily accessible building blocks. Mixed-graft block copolymers containing sequence-defined side chains were prepared through ring-opening metathesis copolymerization of three or four types of macromonomers. Intramolecularly defined interfaces promoted the formation of ordered hierarchical structures with lattice sizes tunable across multiple length scales. The same set of macromonomers were arranged and combined in different ways, providing access to diverse morphologies in the resultant structures.

E3 ubiquitin ligase TRIM29 promotes pancreatic cancer growth and progression via stabilizing Yes-associated protein 1.

BACKGROUND: Pancreatic cancer (PC) is one of the most fatal digestive system cancers. tripartite motif-29 (TRIM29) has been reported as oncogene in several human cancers. However, the precise role and underlying signal cascade of TRIM29 in PC progression remain unclear. METHODS: Western blot, qRT-PCR and immunohistochemistry were used to analyze TRIM29 and Yes-associated protein 1 (YAP1) levels. CCK8 assays, EdU assays and flow cytometry were designed to explore the function and potential mechanism of TRIM29 and YAP1 in the proliferation of PC. Next, a nude mouse model of PC was established for validating the roles of TRIM29 and YAP1 in vivo. The relationship among TRIM29 and YAP1 was explored by co-immunoprecipitation and in vitro ubiquitination assay. RESULTS: TRIM29 and YAP1 was significantly upregulated in PC patient samples, and TRIM29 expression was closely related to a malignant phenotype and poorer overall survival (OS) of PC patients. Functional assays revealed that TRIM29 knockdown suppresses cell growth, arrests cell cycle progression and promotes cell apoptosis of PC cells in vivo and in vitro. Furthermore, the rescue experiments demonstrated that TRIM29-induced proliferation is dependent on YAP1 in PC cells. Mechanistically, TRIM29 regulates YAP1 expression by directly binding to YAP1, and reduced its ubiquitination and degradation. CONCLUSION: Taken together, these results identify a novel mechanism used by PC growth, and provide insight regarding the role of TRIM29 in PC.

Crumpled Versus Flat Gold Nanosheets: Temperature-Regulated Synthesis and Their Plasmonic and Catalytic Properties.

We report a high-yield synthesis of gold (Au) nanosheets with tunable size and surface morphology in the aqueous phase. In particular, crumpled and flat Au nanosheets with a thickness of ∼10 nm could be selectively produced in high purity when the reaction was conducted at room temperature and in an ice-water bath, respectively. Unlike Au nanoplates/nanoprisms in the form of well-defined triangles or hexagons documented in previous studies, the current products exhibit random in-plane branches or holes, together with wavy edges. Strong absorbance in the NIR region was observed for all the Au nanosheet products. When serving as electrocatalysts for the ethanol oxidation reaction, the current products exhibited an enhanced activity and operation stability, as compared to quasi-spherical counterparts.

MicroRNA-148a-3p suppresses epithelial-to-mesenchymal transition and stemness properties via Wnt1-mediated Wnt/ß-catenin pathway in pancreatic cancer.

Although miR-148a-3p has been reported to function as a tumour suppressor in various cancers, the molecular mechanism of miR-148a-3p in regulating epithelial-to-mesenchymal transition (EMT) and stemness properties of pancreatic cancer (PC) cells remains to be elucidated. In the present study, we demonstrated that miR-148a-3p expression was remarkably down-regulated in PC tissues and cell lines. Moreover, low expression of miR-148a-3p was associated with poorer overall survival (OS) in patients with PC. In vitro, gain-of-function and loss-of-function experiments showed that miR-148a-3p suppressed EMT and stemness properties as well as the proliferation, migration and invasion of PC cells. A dual-luciferase reporter assay demonstrated that Wnt1 was a direct target of miR-148a-3p, and its expression was inversely associated with miR-148a-3p in PC tissues. Furthermore, miR-148a-3p suppressed the Wnt/ß-catenin pathway via down-regulation of Wnt1. The effects of ectopic miR-148a-3p were rescued by Wnt1 overexpression. These biological functions of miR-148a-3p in PC were also confirmed in a nude mouse xenograft model. Taken together, these findings suggest that miR-148a-3p suppresses PC cell proliferation, invasion, EMT and stemness properties via inhibiting Wnt1-mediated Wnt/ß-catenin pathway and could be a potential prognostic biomarker as well as a therapeutic target in PC.

Seed-Morphology-Directed Synthesis of Concave Gold Nanocrystals with Tunable Sizes.

We report the fabrication of concave gold (Au) nanocrystals with a set of morphologies and controlled sizes via seeded growth. Starting with Au seeds with a well-defined morphology and uniform size, cubic and rodlike Au nanocrystals with a noticeable concave feature could be successfully obtained, respectively. We also track the growth process and record the shape evolution process. The effect of several reaction parameters on product morphology, such as capping agent and concentration of Ag+, are systematically investigated. Their optical and electrochemical properties are investigated via UV-vis extinction spectroscopy and cyclic voltammetry, respectively. Compared to spherical counterparts, the current concave Au nanocrystals exhibit a noticeable red shift of the absorbance peak in UV-vis extinction spectra and characterized electrochemical behavior of stepped facets, illustrating the morphological advantage.

Serum bile acids profiling by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and its application on pediatric liver and intestinal diseases.

Background A method for bile acid profiling measuring 21 primary and secondary bile acids in serum samples was developed and validated with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Sample preparation included spiking with internal standards followed by protein precipitation, centrifugation, drying under nitrogen gas and reconstitution. Extracted samples were injected onto a Phenomenex Kinetex C18 column (150 × 4.60 mm, 2.6 µm). Methods Data was collected with LC-MS/MS operated in negative ion mode with multiple reaction monitoring (MRM) and single reaction monitoring (SRM). The analytical run time was 12 min. Results The method showed excellent linearity with high regression coefficients (>0.99) over a range of 0.05 and 25 µM for all analytes tested. The method also showed acceptable intra-day and inter-day accuracy and precision. As a proof of concept, the analytical method was applied to patients with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD), biliary atresia (BA), and necrotizing enterocolitis (NEC), and distinct bile acids profiles were demonstrated. Conclusions The method could be poised to identify possible biomarkers for non-invasive early diagnosis of these disorders.

TBRG4 silencing promotes progression of squamous cell carcinoma via regulation of CAV-1 expression and ROS formation.

Esophageal cancer is the eighth most common cancer globally. Transforming growth factor ß regulator 4 (TBRG4) and caveolin-1 (CAV-1) are implicated in tumor progression. The aim of this study was to investigate the expressions of TBRG4 and CAV-1 in esophageal squamous cell carcinoma (ESCC), and their relationship with reactive oxygen species (ROS) formation. Human ESCC cell lines (EC9706, TE-1, and Eca109), and normal esophageal mucosal cell line (Het-1) were used in this study. The silencing of TBRG4 and/or CAV-1 by sh-RNA or overexpression of CAV-1 after TBRG4 knockdown was used to assess ROS levels. The results showed that down-regulation of TBRG4 reduced CAV-1 expression, and promoted ROS formation in ESCCs (p < 0.01). However, CAV-1 overexpression increased the expression level of TBRG4, but decreased ROS level in EC9706 cells (p < 0.01). Similarly, TBRG4 knockdown significantly reduced CAV-1 expression, promoted ROS formation, and caused cell cycle arrest at G0/G1 phase (p < 0.01). Caveolin-1 (CAV-1) knockdown also promoted cell apoptosis, cellular ROS formation and cell cycle arrest at G0/G1 phase (p < 0.01). However, CAV-1 overexpression in sh-TBRG4-treated EC9706 cells significantly upregulated TBRG4 expression, but significantly reduced the level of ROS, and inhibited cell-cycle arrest and apoptosis (p < 0.01). The enhancements in bcl-2/bax ratio, cytochrome c expression, and ROS levels by sh-TBRG4 were significantly reversed by CAV-1 overexpression in EC9706 cells. These results show that the upregulated expression of TBRG4 or CAV-1 promotes ESCC progression via regulation of intracellular ROS levels and inhibition of mitochondria-dependent apoptotic pathway.