Liensinine inhibits progression of intrahepatic cholangiocarcinoma by regulating TGF-ß1 /P-smad3 signaling through HIF-1a.

Intrahepatic cholangiocarcinoma (ICC) is a high-grade malignant digestive system tumor with an insidious onset and unfavorable prognosis. Liensinine, a small molecule derived from plants, has been proven to have significant tumor suppressor activity in other cancers. However, there are no reports on whether liensinine can inhibit the proliferation or metastasis of ICC. This study aimed to explore the tumor-suppressive activity of liensinine in ICC and its underlying mechanisms. The phenotypic changes in ICC cells were monitored in vitro using cell function tests. Western blot and immunofluorescence analyses verified the efficacy of liensinine. Tumor-bearing nude mice were used to explore the effect of liensinine on tumors and its toxicity and side effects in vivo. Liensinine suppressed ICC cell proliferation and arrested the cell cycle at the G1 phase. The epithelial-mesenchymal transition (EMT) of ICC cells was also inhibited, thereby restraining their invasion and migration of tumor cells. In addition, this study found that the potential mechanism of liensinine inhibiting EMT may be via suppression of the TGF-ß1/P-smad3 signaling pathway through hypoxia-inducible factor 1 alpha (HIF-1a). In vivo experiments showed that liensinine inhibited the growth of Hucc-T1 transplanted tumors in nude mice. Liensinine restrained the proliferation of ICC cells and suppressed EMT in ICC via the HIF-1a-mediated TGF-ß1/P-smad3 signaling pathway.

Construction of a Z-scheme heterojunction bifunctional photocatalyst with Ag-modified AgBr embedded in ß-Bi2O3 flowers.

ß-Bi2O3 demonstrates excellent photocatalytic activity under visible light, but it has a very high photogenerated e--h+ recombination rate and quite low quantum efficiency. AgBr also shows excellent catalytic activity but Ag+ is easily reduced to Ag under light radiation, which limits its application in the photocatalysis field, and there are few reports about the application of AgBr in photocatalysis. In this study, the spherical flower-like porous ß-Bi2O3 matrix was first obtained, and then the spherical-like AgBr was embedded between the petals of the flower-like structure to avoid direct light radiation. The only light through the pores on the ß-Bi2O3 petals could be transmitted onto the surfaces of AgBr particles to form a nanometer point light source, which photo-reduced Ag+ on the surface of the AgBr nanospheres to construct the Ag-modified AgBr/ß-Bi2O3 embedded composite and a typical Z-scheme heterojunction was constructed. Under this bifunctional photocatalyst and visible light, the RhB degradation rate reached 99.85% in 30 min, and the photolysis water hydrogen production rate reached 6.288 mmol g-1 h-1. This work is as an effective method for not only the preparation of the embedded structure, quantum dot modification and flower-like morphology but also for the construction of Z-scheme heterostructures.

Active secretion of a thermostable transglutaminase variant in Escherichia coli.

BACKGROUND: Streptomyces mobaraenesis transglutaminase (smTG) is widely used to generate protein crosslinking or attachment of small molecules. However, the low thermostability is a main obstacle for smTG application. In addition, it is still hard to achieve the secretory expression of active smTG in E. coli, which benefits the enzyme evolution. In this study, a combined strategy was conducted to improve the thermostability and secretory expression of active smTG in E. coli. RESULTS: First, the thermostable S. mobaraenesis transglutaminase variant S2P-S23V-Y24N-S199A-K294L (TGm1) was intracellularly expressed in pro-enzyme form in E. coli. Fusing the pro-region of Streptomyces hygroscopicus transglutaminase (proH) and TrxA achieved a 9.78 U/mL of intracellular smTG activity, 1.37-fold higher than the TGm1 fused with its native pro-region. After in vitro activation by dispase, the TGm1 with proH yielded FRAPD-TGm1, exhibiting 0.95 â„ƒ and 94.25% increases in melting temperature and half-life at 60 â„ƒ compared to FRAP-TGm1 derived from the expression using its native pro-region, respectively. Second, the TGm1 with proH was co-expressed with transglutaminase activating protease and chaperones (DnaK, DnaJ, and GrpE) in E. coli, achieving 9.51 U/mL of intracellular FRAPD-TGm1 without in vitro activation. Third, the pelB signal peptide was used to mediate the secretory expression of active TGm in E. coli, yielding 0.54 U/mL of the extracellular FRAPD-TGm1. A script was developed to shuffle the codon of pelB and calculate the corresponding mRNA folding energy. A 1.8-fold increase in the extracellular expression of FRAPD-TGm1 was achieved by the Top-9 pelB sequence derived from the coding sequences with the lowest mRNA folding energy. Last, deleting the gene of Braun's lipoprotein further increased the extracellular yield of FRAPD-TGm1 by 31.2%, reached 1.99 U/mL. CONCLUSIONS: The stabilized FRAPD-smTG here could benefit the enzyme application in food and non-food sectors, while the E. coli system that enables secretory expression of active smTG will facilitate the directed evolution for further improved catalytic properties. The combined strategy (N-terminal modification, co-expression with chaperones, mRNA folding energy optimization of signal peptide, and lipoprotein deletion) may also improve the secretory expression of other functional proteins in E. coli.

Genome-wide identification of key enzyme-encoding genes and the catalytic roles of two 2-oxoglutarate-dependent dioxygenase involved in flavonoid biosynthesis in Cannabis sativa L.

BACKGROUND: Flavonoids are necessary for plant growth and resistance to adversity and stress. They are also an essential nutrient for human diet and health. Among the metabolites produced in Cannabis sativa (C. sativa), phytocannabinoids have undergone extensive research on their structures, biosynthesis, and biological activities. Besides the phytocannabinoids, C. sativa is also rich in terpenes, alkaloids, and flavonoids, although little research has been conducted in this area. RESULTS: In this study, we identified 11 classes of key enzyme-encoding genes, including 56 members involved in the flavonoid biosynthesis in C. sativa, from their physical characteristics to their expression patterns. We screened the potentially step-by-step enzymes catalyzing the precursor phenylalanine to the end flavonoids using a conjoin analysis of gene expression with metabolomics from different tissues and chemovars. Flavonol synthase (FLS), belonging to the 2-oxoglutarate-dependent dioxygenase (2-ODD) superfamily, catalyzes the dihydroflavonols to flavonols. In vitro recombinant protein activity analysis revealed that CsFLS2 and CsFLS3 had a dual function in converting naringenin (Nar) to dihydrokaempferol (DHK), as well as dihydroflavonols to flavonols with different substrate preferences. Meanwhile, we found that CsFLS2 produced apigenin (Api) in addition to DHK and kaempferol when Nar was used as the substrate, indicating that CsFLS2 has an evolutionary relationship with Cannabis flavone synthase I. CONCLUSIONS: Our study identified key enzyme-encoding genes involved in the biosynthesis of flavonoids in C. sativa and highlighted the key CsFLS genes that generate flavonols and their diversified functions in C. sativa flavonoid production. This study paves the way for reconstructing the entire pathway for C. sativa's flavonols and cannflavins production in heterologous systems or plant culture, and provides a theoretical foundation for discovering new cannabis-specific flavonoids.

Common-mode noise self-suppressed 3-component fiber optic accelerometer based on low-reflectivity Bragg gratings.

Common-mode noises (CMNs) are frequent in the fiber optic accelerometer, and their suppression is extremely important, particularly in the ultra-weak signal detection application, e.g., micro-seismic monitoring. This Letter proposes a 3-component (3C) low-reflectivity fiber Bragg gratings accelerometer for CMN self-suppression. When compared with the traditional CMN suppression method, the proposed 3C accelerometer is able to improve the CMN suppression effect by an average value larger than 4.5 dB in three axes, as well as double the effective signal amplitude due to the push-pull structure, which brings an enhanced signal-to-noise ratio. Besides, the proposed 3C accelerometer does not need an additional reference interferometer to achieve such a CMN suppression effect; hence, it largely reduces the volume and cost of the sensing system, which shows huge advantages, particularly in the large-scale quasi-distributed array. The proposed 3C accelerometer provides a promising candidate for the weak vector vibration detection.

Risk and protective factors for chronic pain following inguinal hernia repair: a retrospective study.

PURPOSE: A large proportion of patients experience chronic post-surgical pain (CPSP) following inguinal hernia repair surgery. The aim of this study was to investigate the predictive risk factors and protective factors for CPSP following inguinal hernia surgery. METHODS: After institutional ethics approval was obtained, we conducted a retrospective observational case-control study including a total of 236 adult patients undergoing elective inguinal hernia repair at a single tertiary medical center from 2014 to 2015. Preoperative and postoperative variables were collected from electronic medical records. Binary logistic analysis was used to determine the association between CPSP and clinical factors and built a CPSP risk model. RESULTS: The incidence of CPSP was 14.4%. Bilateral inguinal hernia repair (OR 4.44; 95% CI 1.62 to 12.17; p = 0.004), preoperative pain (OR 2.57; 95% CI 1.14 to 5.79; p = 0.023), preoperative anxiety (OR 1.05; 95% CI 1.01 to 1.09; p = 0.018), and relatively high intensity of acute pain at 1 week after the surgery (OR 1.40; 95% CI 1.03 to 1.91; p = 0.031) were the risk factors for CPSP while low-dose ketamine at anesthesia induction (OR 0.42; 95% CI 0.18 to 0.98; p = 0.044) was the protective factor for CPSP in patients undergoing inguinal hernia repair. CONCLUSIONS: These results indicated that bilateral inguinal hernia repair, preoperative pain, preoperative anxiety, and acute pain at 1 week after the surgery were the independent risk factors for CPSP while low-dose ketamine was the protective factor. These findings may assist with primary prevention by allowing clinicians to screen for individuals with the risk of CPSP.

Multiple OAM vortex beams generation using 1-bit metasurface.

In this paper, a novel method to generate multiple orbital angular momentum (OAM) vortex beams is proposed using a 1-bit metasurface. Through carefully adjusting the phase shift of each element, mirror-symmetrical OAM vortex beams are generated in targeted directions by a 1-bit metasurface under plane-wave illumination. Moreover, the topological charges of the generated vortex beams are opposite. Based on this phenomenon, dual-beam, four-beam, and full-space OAM vortex waves are respectively studied and generated by conducting full-wave simulations. Finally, a prototype of the proposed metasurface is fabricated and measured in an anechoic chamber. The measurement results show that multiple OAM vortex beams are successfully generated and detected, verifying the effectiveness of the proposed method.

Synthesis of core-shell silicon-carbon nanocomposites via in-situ molten salt-based reduction of rice husks: A promising approach for the manufacture of lithium-ion battery anodes.

Silicon (Si) has gained substantial interest as a potential component of lithium-ion battery (LIB) anodes due to its high theoretical specific capacity. However, conventional methods for producing Si for anodes involve expensive metal reductants and stringent reducing environments. This paper describes the development of a calcium hydride (CaH2)-aluminum chloride (AlCl3) reduction system that was used for the in-situ low-temperature synthesis of a core-shell structured silicon-carbon (Si-C) material from rice husks (RHs), and the material was denoted RHs-Si@C. Moreover, as an LIB anode, RHs-Si@C exhibited exceptional cycling performance, exemplified by 90.63 % capacity retention at 5 A g-1 over 2000 cycles. Furthermore, the CaH2-AlCl3 reduction system was employed to produce Si nanoparticles (Si NPs) from RHs (R-SiO2, where SiO2 is silica) and from commercial silica (C-SiO2). The R-SiO2-derived Si NPs exhibited a higher residual silicon oxides (SiOx) content than the C-SiO2-derived Si NPs. This was advantageous, as there was sufficient SiOx in the R-SiO2-derived Si NPs to mitigate the volumetric expansion typically associated with Si NPs, resulting in enhanced cycling performance. Impressively, Si NPs were fabricated on a kilogram scale from C-SiO2 in a yield of 82 %, underscoring the scalability of the low-temperature reduction technique.

Prognostic value of diffuse reduction of spleen density on postoperative survival of pancreatic ductal adenocarcinoma: A retrospective study.

PURPOSE: It is difficult to predict the prognosis of patients with pancreatic ductal adenocarcinoma (PDAC) before radical operation. The purpose of this study was to explore the connection between the diffuse reduction of spleen density on computed tomography (DROSD) and the postoperative prognosis of patients with PDAC. PATIENTS AND METHODS: A total of 160 patients with PDAC who underwent radical surgery in the First Affiliated Hospital of Wenzhou Medical University were enrolled. Cox regression analysis was used to cast the overall survival (OS) and evaluate the prognostic factors. Nomogram was used to forecast the possibility of 1-year, 3-year, and 5-year OS. The prediction accuracy and clinical net benefit are performed by concordance index (C-index), calibration curve, time-dependent receiver operating characteristics (tdROC), and decision curve analysis. RESULTS: In multivariable Cox analysis, DROSD is independently related to OS. Advanced age, TNM stage, neutrophil/lymphocyte ratio, and severe complications were also independent prognostic factors. The calibration curves of nomogram showed optimal agreement between prediction and observation. The C-index of nomogram is 0.662 (95%CI, 0.606-0.754). The area under tdROC curve for a 3-year OS of nomogram is 0.770. CONCLUSION: DROSD is an independent risk factor for an OS of PDAC. We developed a nomogram that combined imaging features, clinicopathological factors, and systemic inflammatory response to provide a personalized risk assessment for patients with PDAC.

Characterization and co-expression analysis of ATP-binding cassette transporters provide insight into genes related to cannabinoid transport in Cannabis sativa L.

Plant ATP-binding cassette (ABC) transporters contribute the transport of diverse secondary metabolites. However, their roles in cannabinoid trafficking are still unsolved in Cannabis sativa. In this study, 113 ABC transporters were identified and characterized in C. sativa from their physicochemical properties, gene structure, and phylogenic relationship, as well as spatial gene expression patterns. Eventually, seven core transporters were proposed including one member in ABC subfamily B (CsABCB8) and six ABCG members (CsABCG4, CsABCG10, CsABCG11, CsABCG32, CsABCG37, and CsABCG41), harboring potential in participating cannabinoid transport, by combining phylogenetic and co-expression analysis from the gene and metabolite level. The candidate genes exhibited a high correlation with cannabinoid biosynthetic pathway genes and the cannabinoid content, and they were highly expressed where cannabinoids appropriately biosynthesized and accumulated. The findings underpin further research on the function of ABC transporters in C. sativa, especially in unveiling the mechanisms of cannabinoid transport to boost systematic and targeted metabolic engineering.

AZGP1 activation by lenvatinib suppresses intrahepatic cholangiocarcinoma epithelial-mesenchymal transition through the TGF-ß1/Smad3 pathway.

Intrahepatic cholangiocarcinoma (ICC) is a primary liver malignancy and is characterized by highly aggressive and malignant biological behavior. Currently, effective treatment strategies are limited. The effect of lenvatinib on ICC is unknown. In this study, we found that AZGP1 was the key target of lenvatinib in ICC, and its low expression in ICC cancer tissues was associated with a poor prognosis in patients. Lenvatinib is a novel AZGP1 agonist candidate for ICC that inhibits ICC-EMT by regulating the TGF-ß1/Smad3 signaling pathway in an AZGP1-dependent manner. Furthermore, we found that lenvatinib could increase AZGP1 expression by increasing the acetylation level of H3K27Ac in the promoter region of the AZGP1 gene, thereby inhibiting EMT in ICC cells. In conclusion, lenvatinib activates AZGP1 by increasing the acetylation level of H3K27Ac on the AZGP1 promoter region and regulates the TGF-ß1/Smad3 signaling pathway in an AZGP1-dependent manner to inhibit ICC-EMT. This study offers new insight into the mechanism of lenvatinib in the treatment of ICC and provides a theoretical basis for new treatment methods.

Combining CRISPR-Cpf1 and Recombineering Facilitates Fast and Efficient Genome Editing in Escherichia coli.

Clustered regularly interspaced short palindromic repeat (CRISPR)-based gene-editing technology has been widely used in various microorganisms due to its advantages of low cost, high efficiency, easy operation, and multiple functions. In this study, an efficient and fast double-plasmid gene-editing system pEcCpf1/pcrEG was constructed in Escherichia coli based on CRISPR/Cpf1. First, gene knockout and integration efficiency were verified in eight different kinds of protospacer adjacent motif (PAM) regions. Then, the transformation method was optimized, and the efficiency of gene knockout or gene integration of this system increased to nearly 100%, and the large-length fragments could be integrated into the genome in E. coli BL21 (DE3). The system was also optimized by replacing the homologous recombination system in plasmid pEcCpf1, resulting in pEcCpf1H, which could perform precise single-point mutation, terminator insertion, short-sequence insertion, or gene knockout with high efficiency using a 90 nt (nucleotide) single-stranded primer. Further, multiple genes could be edited simultaneously. Next, these two systems were demonstrated in other E. coli strains. Finally, as an application, the system was used to engineer the synthesis pathway of l-histidine in the engineered strain. The titer of l-histidine in a shake flask reached 7.16 g/L, a value increased by 84.1% compared to the starting strain. Thus, this study provided an effective tool for metabolic engineering of E. coli.

Genome-Wide Identification and Expression Profiles of bZIP Genes in Cannabis sativa L.

Background: The bZIP gene family plays roles in biotic and abiotic stress, secondary metabolism, and other aspects in plants. They have been reported in Arabidopsis thaliana, Oryza sativa, Artemisia annua, and other plants, but their roles in Cannabis sativa have not been determined. Materials and Methods: In this study, we analyzed the genome-wide identification and expression profile of the bZIP gene family in C. sativa. Results: A total of 51 members of the bZIP gene family were identified based on the C. sativa genome and numbered in order from CsbZIP1 to CsbZIP51. Their phylogenetic relationships, cis-elements in promoter region, gene structures and motif compositions, physicochemical properties, chromosome locations, and expression profiles, were analyzed. The results showed that the 51 CsbZIPs were unevenly distributed on 10 chromosomes and could be clustered into 11 subfamilies. Furthermore, CsbZIPs located in the same subfamilies presented similar intron/exon organization and motif composition. The expression levels of CsbZIPs in various tissues (flowers, bracts, vegetative leaves, stems, and seeds) were determined using reverse transcription quantitative polymerase chain reaction. The expression levels of CsbZIPs were higher in flowers and bracts. The 51 CsbZIPs were explored, and their structure, evolution, and expression pattern in different tissues of C. sativa were characterized synthetically. The findings indicated that CsbZIPs are essential for the growth and development of C. sativa. Conclusions: These results provide a theoretical basis for subsequent research on hemp bZIP transcription factors and the cultivation of high-cannabidiol and low-tetrahydrocannabinol high-quality cannabis varieties.

Benefits and risks of intermittent bolus erector spinae plane block through a catheter for patients after cardiac surgery through a lateral mini-thoracotomy: A propensity score matched retrospective cohort study.

STUDY OBJECTIVE: A lateral mini-thoracotomy approach to cardiac surgery causes severe and complicated postoperative pain compared to the sternotomy approach. In this study we assessed the benefits and risks of intermittent bolus erector spinae plane block (ESPB) via a catheter for patients who underwent cardiac surgery through a lateral mini-thoracotomy. DESIGN: A propensity score-matched retrospective cohort study. SETTING: University hospital. PATIENTS: 452 consecutive patients that underwent cardiac surgery through a lateral mini-thoracotomy from 2018 to 2020. INTERVENTIONS: Patients who received intermittent bolus ESPB through a catheter for 3 days (ESPB group, n = 93) were compared with patients who did not receive any regional anesthesia (Control group, n = 174) after propensity score matching. MEASUREMENTS: The primary endpoint was postoperative in-hospital cumulative opioid consumption (calculated as oral morphine milligram equivalents, MME). The secondary outcomes were intraoperative sufentanil doses, therapeutic use of antiemetic, pulmonary infection (assessed using a modified clinical pulmonary infection score, CPIS), durations of ICU and hospital stays, and ESPB related/unrelated complications. MAIN RESULTS: There is a lower oral MME in the ESPB group, 266 ± 126 mg in the ESPB group vs. 346 ± 105 mg in the control group (95% CI -113 to -46; P < 0.01). Fewer patients received therapeutic antiemetic agents in the ESPB group (30% vs. 42%, odds ratio 0.58; 95% CI 0.34 to 0.99; P = 0.04). The modified CPIS in the ESPB group is lower: 1.4 ± 0.9 vs. 2.0 ± 1.0 (95% CI -0.9 to -0.3; P < 0.01) on postoperative day 1; 1.6 ± 0.9 vs. 2.0 ± 0.9 (95% CI -0.7 to -0.2; P < 0.01) on postoperative day 2. The observed complications associated with ESPB include pneumothorax (1%), staxis around stomas (5%), hypotension (1%), catheter displacement (3%), and catheter obstruction (2%). None of the patients had any adverse outcomes. CONCLUSION: Intermittent bolus ESPB is relatively safe and correlated with a reduction in the use of opioids and antiemetics for cardiac surgery through a lateral mini-thoracotomy.

An ortho-rhom-bic polymorph of 1-benzyl-1H-benzimidazole.

The title compound, C(14)H(12)N(2), in contrast to the previously reported monoclinic polymorph [Lei et al. (2009 ▶). Acta Cryst. E65, o2613], crystallizes in the ortho-rhom-bic crystal system. The dihedral angle between the imidazole ring system and the phenyl ring is 76.78 (16)°. Weak C-H⋯N and C-H⋯π inter-actions are observed in the crystal structure.

1-Chloro-methyl-1H-1,2,3-benzotriazole.

In the title compound, C(7)H(6)ClN(3), the benzotriazole ring is essentially planar with a maximum deviation of 0.0110 (15)Å, and makes a dihedral angle of 0.46 (8)° with the benzene ring. In the crystal, mol-ecules are linked through inter-molecular C-H⋯N hydrogen bonds, forming chains along the c axis.

1-(But-2-enyl-idene)-2-(2-nitro-phen-yl)hydrazine.

The mol-ecule of the title Schiff base compound, C(10)H(11)N(3)O(2), adopts an E geometry with respect to the C=N double bond. The mol-ecule is roughly planar, with the largest deviation from the mean plane being 0.111 (2) Å, The enyl-idene-hydrazine group is, however, slightly twisted with respect to the phenyl ring, making a dihedral angle of 6.5 (3)°. An intra-molecular N-H⋯O hydrogen bond may be responsible for the planar conformation. An inter-molecular N-H⋯O hydrogen bond links two mol-ecules around an inversion center, building a pseudo dimer.

2-Eth-oxy-4-{[(2-nitro-phen-yl)hydrazono]meth-yl}phenol.

The title compound, C(15)H(15)N(3)O(4), a Schiff base, was obtained from a condensation reaction of 3-eth-oxy-4-hydroxy-benzaldehyde and 2-nitro-phenyl-hydrazine. The mol-ecule is approximately planar, the largest deviation from the mean plane being 0.1449 (16) Å. An intramolecular N-H⋯O inter-action is also present. In the crystal, inter-molecular O-H⋯O hydrogen bonds link the mol-ecules, forming chain parallel to the b axis.

[µ-1,1'-(Butane-1,4-di-yl)di-1H-benz-imidazole-κN:N]bis-{[N,N'-bis(car-boxy-meth-yl)ethyl-enediamine-N,N'-di-acetato-κO,O',O'',N,N']mercury(II)} methanol disolvate.

The binuclear title complex, [Hg(2)(C(10)H(14)N(2)O(8))(2)(C(18)H(18)N(4))]·2CH(3)OH, lies on an inversion center with the unique Hg(II) ion coordinated in a disorted octa-hedral environment with one Hg-N bond significantly shorter than the other two. In the crystal structure, inter-molecular O-H⋯O hydrogen bonds link complex and solvent mol-ecules into a three-dimensional network.

{4-Bromo-2-[2-(piperidin-1-ium-1yl)ethyl-iminometh-yl]phenolato}diiodido-zinc(II).

In the title complex, [ZnI(2)(C(14)H(19)BrN(2)O)], the Zn(II) atom is four-coordinated by the imine N and phenolate O atoms of the Schiff base ligand and by two iodide ions in a distorted tetra-hedral coordination. In the crystal structure, mol-ecules are linked through inter-molecular N-H⋯O hydrogen bonds, forming chains running along the b axis.