BCLAF1 binds SPOP to stabilize PD-L1 and promotes the development and immune escape of hepatocellular carcinoma.

Interaction between programmed death-1 (PD-1) ligand 1 (PD-L1) on tumor cells and PD-1 on T cells allows tumor cells to evade T cell-mediated immune surveillance. Strategies targeting PD-1/PD-L1 have shown clinical benefits in a variety of cancers. However, limited response rates in hepatocellular carcinoma (HCC) have prompted us to investigate the molecular regulation of PD-L1. Here, we identify B cell lymphoma-2-associated transcription factor 1 (BCLAF1) as a key PD-L1 regulator in HCC. Specifically, BCLAF1 interacts with SPOP, an E3 ligase that mediates the ubiquitination and degradation of PD-L1, thereby competitively inhibiting SPOP-PD-L1 interaction and subsequent ubiquitination and degradation of PD-L1. Furthermore, we determined an SPOP-binding consensus (SBC) motif mediating the BCLAF1-SPOP interaction on BCLAF1 protein and mutation of BCLAF1-SBC motif disrupts the regulation of the SPOP-PD-L1 axis. In addition, BCLAF1 expression was positively correlated with PD-L1 expression and negatively correlated with biomarkers of T cell activation, including CD3 and CD8, as well as with the level of immune cell infiltration in HCC tissues. Besides, BCLAF1 depletion leads to a significant reduction of PD-L1 expression in vitro, and this reduction of PD-L1 promoted T cell-mediated cytotoxicity. Notably, overexpression of BCLAF1 sensitized tumor cells to checkpoint therapy in an in vitro HCC cells-Jurkat cells co-culture model, whereas BCLAF1-SBC mutant decreased tumor cell sensitivity to checkpoint therapy, suggesting that BCLAF1 and its SBC motif serve as a novel therapeutic target for enhancing anti-tumor immunity in HCC.

Pan-Cancer Analysis Identifies BCLAF1 as a Potential Biomarker for Renal Cell Carcinoma.

B-cell lymphoma-2-associated transcription factor 1 (BCLAF1) is a versatile protein involved in the regulation of gene transcription and post-transcriptional processing. Although BCLAF1 exerts a broad tumor suppressor effect or tumor promoter effect in many cancer types, the specific roles concerning its expression levels, and its impact on tumorigenesis in Renal cell carcinoma (RCC) remain unclear. Here, we utilized the Cancer Genome Atlas (TCGA) and Genotype Tissue Expression (GTEx) datasets alongside R software and online tools to unravel the specific roles of BCLAF1 in 33 cancer types, including its expression levels, tumor immune and molecular subtypes, and its correlation with prognosis, diagnosis, DNA methylation, and immune microenvironment. Additionally, we carried out cell biology experiments to independently investigate the expression of BCLAF1 in RCC and its effects on tumor progression. BCLAF1 was differentially expressed in tumor tissues compared to normal tissues across various cancer types and was also differentially expressed in different immune and molecular subtypes. In RCC, patients with high BCLAF1 expression had a better prognosis and BCLAF1 was tightly correlated with the stage, gender, and histological grade of patients. Furthermore, BCLAF1 had higher DNA methylation levels and higher immune infiltration levels in tumor tissues. Additionally, cell functional experiments confirmed the low expression of BCLAF1 in RCC and that BCLAF1 significantly inhibited the proliferation, migration, and invasion, while inducing apoptosis and cell cycle arrest in RCC cells in vitro. Our study under-scored the potential of BCLAF1 as an important actor in tumorigenesis, especially concerning RCC where it may serve as an effective prognostic marker.

Distinct hybridization modes in wide- and narrow-ranged lineages of Causonis (Vitaceae).

BACKGROUND: Explaining contrasting patterns of distribution between related species is crucial for understanding the dynamics of biodiversity. Despite instances where hybridization and whole genome duplication (WGD) can yield detrimental outcomes, a role in facilitating the expansion of distribution range has been proposed. The Vitaceae genus Causonis exhibits great variations in species' distribution ranges, with most species in the derived lineages having a much wider range than those in the early-diverged lineages. Hybridization and WGD events have been suggested to occur in Causonis based on evidence of phylogenetic discordance. The genus, therefore, provides us with an opportunity to for explore different hybridization and polyploidization modes in lineages with contrasting species' distribution ranges. However, the evolutionary history of Causonis incorporating potential hybridization and WGD events remains to be explored. RESULTS: With plastid and nuclear data from dense sampling, this study resolved the phylogenetic relationships within Causonis and revealed significant cyto-nuclear discordance. Nuclear gene tree conflicts were detected across the genus, especially in the japonica-corniculata clade, which were mainly attributed to gene flow. This study also inferred the allopolyploid origin of the core Causonis species, which promoted the accumulation of stress-related genes. Causonis was estimated to have originated in continental Asia in the early Eocene, and experienced glaciation in the early Oligocene, shortly after the divergence of the early-divergent lineages. The japonica-corniculata clade mainly diversified in the Miocene, followed by temperature declines that may have facilitated secondary contact. Species distribution modeling based on current climate change predicted that the widespread C. japonica tends to be more invasive, while the endemic C. ciliifera may be at risk of extinction. CONCLUSIONS: This study presents Causonis, a genus with complex reticulate evolutionary history, as a model of how hybridization and WGD modes differ in lineages of contrasting species' geographic ranges. It is important to consider specific evolutionary histories and genetic properties of the focal species within conservation strategies.

Who will name new plant species? Temporal change in the origins of taxonomists in China.

Discovery rates of new plant species need to be accelerated because many species will be extinct before they are formally described. Current studies have focused on where new species may occur and their characteristics. However, who will actually discover and describe these new species has received limited attention. Here, we used 31 576 vascular plant species distributed and described in China as a case study to explore the temporal patterns of the nationalities of the taxonomists. We found that most recently described species are endemic species, and there has been an increasing proportion of species descriptions by resident Chinese taxonomists over time. The proportion of species described by resident taxonomists reached an average of 80.8% between 1977 and 2018. By contrast, species discoveries by non-resident experts, often non-endemic species, showed signs of levelling off. Our study underscores an urgent need for training of, support for and collaboration with resident taxonomists in megadiverse countries with a high potential of discovering undescribed plant species.

Endometrial cancer (EC) derived G3BP1 overexpression and mutant promote EC tumorigenesis and metastasis via SPOP/ERα axis.

BACKGROUND: Ras-GTPase-activating protein binding protein 1 (G3BP1) is an oncogenic factor, which highly expressed in a variety of cancers. In recent years, G3BP1 has been reported to promote the development of prostate cancer by inhibiting the degradation of AR through inhibiting SPOP. However, whether G3BP1 contributes in a similar manner to the abnormal accumulation of ERα, which is also an important target for hormone therapy, remains unknown. This article addresses this issue and explores potential mechanisms. METHODS: Bioinformatics tools were used for G3BP1 expression analysis, survival analysis, and clinical association analysis. Immunohistochemical staining was used to examine the correlation between G3BP1 and ERα in EC patients. In addition, western blot and co-immunoprecipitation were used to detect the half-life of G3BP1 and mutant, and the effect of G3BP1 and mutant on the ubiquitination and degradation of ERα mediated by SPOP. Then, the oncogenic functions of G3BP1 dependent on the SPOP/ERα axis were determined by CCK8 cell proliferation assay, colony formation assay and cell migration assay. Finally, we established the EC cells treated or untreated with fulvestrant, exploring the possibility of fulvestrant combined with the reduction of G3BP1 to improve the efficacy of fulvestrant. RESULTS: G3BP1 is abnormally high expressed and characterized by high-frequency mutation in EC. In addition, there is a positive correlation between G3BP1 protein and ERα protein. Mechanistically, both G3BP1 and mutant, the latter is displaying the longer half-life, competitively bind SPOP with ERα, thereby inhibiting SPOP-mediated ubiquitination and degradation of ERα. Functionally, G3BP1 and mutant promote the proliferation and migration of EC cells by regulating the G3BP1/SPOP/ERα axis. However, fulvestrant can reverse the cancer-promoting effects of G3BP1 and mutant. CONCLUSIONS: G3BP1 and its mutant positively regulate ERα signaling pathway by inhibiting SPOP-mediated ubiquitination and degradation of ERα, indicating the promising effect of fulvestrant on the suppression the occurrence and development of EC with high expressed G3BP1 and G3BP1 mutants. Video Abstract.

Prostate cancer-associated SPOP mutations lead to genomic instability through disruption of the SPOP-HIPK2 axis.

Speckle-type Poz protein (SPOP), an E3 ubiquitin ligase adaptor, is the most frequently mutated gene in prostate cancer. The SPOP-mutated subtype of prostate cancer shows high genomic instability, but the underlying mechanisms causing this phenotype are still largely unknown. Here, we report that upon DNA damage, SPOP is phosphorylated at Ser119 by the ATM serine/threonine kinase, which potentiates the binding of SPOP to homeodomain-interacting protein kinase 2 (HIPK2), resulting in a nondegradative ubiquitination of HIPK2. This modification subsequently increases the phosphorylation activity of HIPK2 toward HP1γ, and then promotes the dissociation of HP1γ from trimethylated (Lys9) histone H3 (H3K9me3) to initiate DNA damage repair. Moreover, the effect of SPOP on the HIPK2-HP1γ axis is abrogated by prostate cancer-associated SPOP mutations. Our findings provide new insights into the molecular mechanism of SPOP mutations-driven genomic instability in prostate cancer.

Pharyngeal reconstruction for severe pharyngeal stenosis in Behçet's disease: a retrospective case series.

OBJECTIVES: Pharyngeal scarring stenosis is a rare yet very severe complication in Behçet's disease (BD). Previously, such patients had to undergo tracheostomy for life, which seriously affected the patient's speech and swallowing function. We aim to present the effect of pharyngeal reconstructive surgeries using flaps for severe pharyngeal stenosis in BD. METHODS: The medical history, the surgical procedures of reconstruction and the clinical outcomes of BD cases with pharyngeal stenosis were analysed. British Medical Research Council questionnaire (MRC), Chinese version of the Swallowing Quality-of-Life Questionnaire (SWQOL), the dysphagia score (DS) and the penetration-aspiration scale (PAS) based on videofluoroscopic swallowing study were used. RESULTS: Six BD cases with pharyngeal stenosis underwent reconstructive operations. Submental island flaps and forearm free flaps were used in reconstructive procedures in three female and three male patients, respectively. All patients successfully removed the tracheotomy cannula and nasal feeding tube after reconstruction. Dyspnoea was significantly relived as MRC scores decreased from 3 (3-4) to 1 (1-2) (p=0.020, Z=-2.333). SWQOL scores were remarkably improved from 782.5 (657.0-854.0) to 826.5 (768.0-864.0) (p=0.027, Z=-2.207). There was non-significant decrease in DS (from 2.5 to 1.5, p=0.066, Z=-1.841) and increase in PAS (from 1 to 1.5, p=0.317, Z=-1.000). CONCLUSIONS: Reconstructive surgery using flaps is an effective and safe approach to rebuild pharyngeal cavity in BD patients with severe pharyngeal stenosis, which can improve the quality of life of these patients.

Kelch-like protein 3 in human disease and therapy.

Kelch-like protein 3 (KLHL3) is a substrate adaptor of Cullin3-RING ubiquitin ligase (CRL3), and KLHL3-CUL3 complex plays a vital role in the ubiquitination of specific substrates. Mutations and abnormal post-translational modifications of KLHL3-CUL3 affect substrate ubiquitination and may related to the pathogenesis of Gordon syndrome (GS), Primary Hyperparathyroidism (PHPT), Diabetes Mellitus (DM), Congenital Heart Disease (CHD), Pre-eclampsia (PE) and even cancers. Therefore, it is essential to understand the function and molecular mechanisms of KLHL3-CUL3 for the treatment of related diseases. In this review, we summary the structure and function of KLHL3-CUL3, the effect of KLHL3-CUL3 mutations and aberrant modifications in GS, PHPT, DM, CHD and PE. Moreover, we noted a possible role of KLHL3-CUL3 in carcinogenesis and provided ideas for targeting KLHL3-CUL3 for related disease treatment.

Role of formin INF2 in human diseases.

Formin proteins catalyze actin nucleation and microfilament polymerization. Inverted formin 2 (INF2) is an atypical diaphanous-related formin characterized by polymerization and depolymerization of actin. Accumulating evidence showed that INF2 is associated with kidney disease focal segmental glomerulosclerosis and cancers, such as colorectal and thyroid cancer where it functions as a tumor suppressor, glioblastoma, breast, prostate, and gastric cancer, via its oncogenic function. However, studies on the underlying molecular mechanisms of the different roles of INF2 in diverse cancers are limited. This review comprehensively describes the structure, biochemical features, and primary pathogenic mutations of INF2.

Generation of a 25-line flattened optical frequency comb and its coherent beating properties.

A 25-line flattened optical frequency comb (OFC) is generated by cascading Mach-Zehnder modulators, and its coherent beating properties are investigated. The effect of bias voltages and the power of radio frequency signals on the flatness of the OFC, and the phase relationships between each comb line are theoretically deduced. In the experiment, a 25-line OFC with a flatness of less than 1 dB and an optical sideband suppression ratio of 12 dB is successfully demonstrated, and the agreement between simulation and experimental results supports the validity of the model for analysis of the flatness. The measured amplitude of harmonic signals by coherent beating of the generated 25-line OFC matches well with the theoretical prediction. A phase noise of -105.31dBc/Hz at 10 kHz frequency offset for the 12th harmonic signal almost without phase noise degradation proves the high coherence of the generated OFC lines.

SPOP suppresses testicular germ cell tumors progression through ubiquitination and degradation of DPPA2.

Dysregulation of the ubiquitin-proteasome pathway is strongly associated with cancer initiation and progression. Speckle-type POZ(pox virus and zinc finger protein) protein(SPOP) is an adapter protein of CUL3-based E3 ubiquitin ligase complexes. Gene expression profiling from the Cancer Genome Atlas (TCGA) suggests that SPOP is downregulated in testicular germ cell tumors (TGCTs), but the specific contribution of this protein remains to be explored. In this study, we show that the germ line-specific factor DPPA2 was identified as a proteolytic substrate for the SPOP-CUL3-RBX1 E3 ubiquitin-ligase complex. SPOP specifically binds to a SPOP-binding consensus (SBC) degron located in DPPA2 and targets DPPA2 for degradation via the ubiquitin-proteasome pathway. SPOP downregulation increases the expression of pluripotency markers OCT4 and Nanog but decreases that of early differentiation marker gene Fst. This effect is partly dependent on its activity toward DPPA2. In addition, the dysregulation of SPOP-DPPA2 axis contributes to the malignant transformation phenotypes of TGCT cells.

Photonic generation of terahertz dual-chirp waveforms ranging from 364 to 392†GHz.

In this paper, we propose and experimentally demonstrate a photonic scheme based on frequency doubling and photo-mixing to generate dual-chirp signals in the terahertz (THz) band. A broadband dual-chirp THz signal with 28 GHz bandwidth, ranging from 364 GHz to 392 GHz, is successfully generated in the proof-of-concept experiment, resulting in a chirp rate of 0.028 GHz/ns for both up chirp and down chirp signals. THz dual-chirp signals featuring a large bandwidth are beneficial to enable high resolution and high accuracy by mitigating the range measurement error induced by the range-Doppler coupling effect. Therefore, the proposed system is expected to have a great potential for future THz radar applications.

Spin-to-Charge Conversion in Ag/Bi Bilayer Revisited.

The spin-to-charge conversion of the Ag/Bi interface is studied in a device in which a spin current can be injected from either side selectively. The charge voltages generated by the two counterpropagating spin currents show opposite signs, that is consistent with the inverse spin Hall effect rather than the well-accepted inverse Rashba-Eldestein effect in the Ag/Bi bilayer. Femtosecond laser is further employed to generate the spin-current-induced terahertz signal in a Ag/Bi bilayer, which shows no evidence for the inverse Rashba-Eldestein effect, either. This work provides a clear-cut method to identify the spin-to-charge mechanism in a Rashba electronic state and delivers new understanding for the relevant spin-transport phenomena.

Analysis of reflux as the aetiology of laryngeal dysplasia progression through a matched case-control study.

OBJECTIVES: Laryngeal dysplasia (LD) is a precancerous lesion of the larynx. In this study, the laryngeal tissue of patients with laryngeal dysplasia was taken as the research object, and the aetiology of reflux was analysed. METHOD: Patients with laryngeal dysplasia after surgery were selected as our subjects. The levels of pepsin, enterokinase and bilirubin in laryngeal tissue samples of the two groups were detected by immunohistochemical method. RESULTS: The OR values (95% CI) of pepsin, enterokinase and bilirubin were 0.67 (0.19-2.36), 0.80 (0.22-2.98) and 1.33 (0.30-5.96), respectively, in the univariate analysis. Besides, in the multivariate analysis, the OR values (95% CI) of pepsin, enterokinase and bilirubin were 0.57 (0.14-2.30), 0.73 (0.18-2.92) and 1.40 (0.30-6.53), respectively. CONCLUSION: Larger sample size should be applied to prospective studies on whether reflux is a risk factor for laryngeal cancer.

lncRNA JPX/miR-33a-5p/Twist1 axis regulates tumorigenesis and metastasis of lung cancer by activating Wnt/ß-catenin signaling.

BACKGROUND: MicroRNAs (miRNAs) and Twist1-induced epithelial-mesenchymal transition (EMT) in cancer cell dissemination are well established, but the involvement of long noncoding RNAs (lncRNAs) in Twist1-mediated signaling remains largely unknown. METHODS: RT-qPCR and western blotting were conducted to detect the expression levels of lncRNA JPX and Twist1 in lung cancer cell lines and tissues. The impact of JPX on Twist1 expression, cell growth, invasion, apoptosis, and in vivo tumor growth were investigated in lung cancer cells by western blotting, rescue experiments, colony formation assay, flow cytometry, and xenograft animal experiment. RESULTS: We observed that lncRNA JPX was upregulated in lung cancer metastatic tissues and was closely correlated with tumor size and an advanced stage. Functionally, JPX promoted lung cancer cell proliferation in vitro and facilitated lung tumor growth in vivo. Additionally, JPX upregulated Twist1 by competitively sponging miR-33a-5p and subsequently induced EMT and lung cancer cell invasion. Interestingly, JPX and Twist1 were coordinately upregulated in lung cancer tissues and cells. Mechanically, the JPX/miR-33a-5p/Twist1 axis participated in EMT progression by activating Wnt/ß-catenin signaling. CONCLUSIONS: These findings suggest that lncRNA JPX, a mediator of Twist1 signaling, could predispose lung cancer cells to metastasis and may serve as a potential target for targeted therapy.

Method of fiber transfer delay measurement based on phase quantization and delay synthesis.

A novel method, to the best of our knowledge, of fiber transfer delay (FTD) measurement based on phase quantization and delay synthesis is proposed and demonstrated. By detecting the differential phase shifts of a set of frequency-multiplied RF signals transmission through the fiber link with and without the FTD under the test, the ${2}\pi $2π phase ambiguity problem can be solved. To avoid the phase quantization error near the digital quantization boundary, a self-check and error-correction method is proposed so as to greatly improve the reliability of measurement. In the experiment, the measurement repeatability around 0.018 ps within a period of 80 s is achieved for a back-to-back fiber link, and a test resolution of 0.03 ps is proved with a motorized tunable delay line. The system is available for measurement of a large FTD range up to 100 µs with no dead zone.

Dysregulation of INF2-mediated mitochondrial fission in SPOP-mutated prostate cancer.

Next-generation sequencing of the exome and genome of prostate cancers has identified numerous genetic alternations. SPOP (Speckle-type POZ Protein) was one of the most frequently mutated genes in primary prostate cancer, suggesting SPOP is a potential driver of prostate cancer development and progression. However, how SPOP mutations contribute to prostate cancer pathogenesis remains poorly understood. SPOP acts as an adaptor protein of the CUL3-RBX1 E3 ubiquitin ligase complex that generally recruits substrates for ubiquitination and subsequent degradation. ER-localized isoform of the formin protein inverted formin 2 (INF2) mediates actin polymerization at ER-mitochondria intersections and facilitates DRP1 recruitment to mitochondria, which is a critical step in mitochondrial fission. Here, we revealed that SPOP recognizes a Ser/Thr (S/T)-rich motif in the C-terminal region of INF2 and triggers atypical polyubiquitination of INF2. These ubiquitination modifications do not lead to INF2 instability, but rather reduces INF2 localization in ER and mitochondrially associated DRP1 puncta formation, therefore abrogates its ability to facilitate mitochondrial fission. INF2 mutant escaping from SPOP-mediated ubiquitination is more potent in prompting mitochondrial fission. Moreover, prostate cancer-associated SPOP mutants increase INF2 localization in ER and promote mitochondrial fission, probably through a dominant-negative effect to inhibit endogenous SPOP. Moreover, INF2 is important for SPOP inactivation-induced prostate cancer cell migration and invasion. These findings reveal novel molecular events underlying the regulation of INF2 function and localization, and provided insights in understanding the relationship between SPOP mutations and dysregulation of mitochondrial dynamics in prostate cancer.

Quality of Life in Treating Persistent Neurogenic Dysphagia with Cricopharyngeal Myotomy.

Neurological impairment is an important cause of dysphagia. This study analyzed whether quality of life (QoL) is improved after coblation-assisted endoscopic cricopharyngeal myotomy (CAECPM) for patients with persistent neurological dysphagia who meet the criteria by using the Chinese version of the Swallow Quality-of-Life Questionnaire (CSWAL-QOL). 22 patients with dysphagia for more than 6 months after stroke or lateral skull base surgery were screened. All patients exhibited a poor response to conservative treatment such as swallowing rehabilitation. Videofluoroscopic swallowing studies (VFSS) showed a restricted cricopharyngeal opening. The preoperative CSWAL-QOL score was 377.7 (311.3-493.0) out of 1000; the postoperative score was 641.7 (293.7-758.3) out of 1000; the preoperative median dysphagia frequency was 41.4 (25.7-61.4) out of 100; and the postoperative median score was 64.3 (24.3-80.0). A significant difference was found between preoperative and postoperative scores together with dysphagia frequency (P < 0.05). Among all the variables, laryngeal elevation ability was statistically significantly correlated with efficacy of CAECPM (P = 0.01). These values indicate that quality of life could be improved after CAECPM for patients with persistent neurological dysphagia, who have cricopharyngeal achalasia. The ability of laryngeal elevation has significant influence. The CSWAL-QOL can be used to assess different aspects of the swallow-related quality of life of these patients.

Generation and Detection of Pure Spin Current in an H-Shaped Structure of a Single Metal.

Distinct from all the existing methods for determining the spin Hall angle with bilayers, we have developed a new approach based on the mesoscopic H-shaped structure, which generates and detects pure spin current in a single metal. Using this approach we have unequivocally addressed the long-standing controversy of the magnitude of the spin Hall angle of gold. By exploiting the long spin diffusion length of Cu and the large spin-orbit coupling of Bi, we have realized a very large spin Hall effect in 10 nm Cu films delta doped with 0.15 nm of Bi. This new artificial material can generate a large spin-orbit torque to switch an adjacent Fe layer. We have thus demonstrated new artificial materials with a simultaneously large spin Hall angle and long spin diffusion length.

Nodeless Bulk Superconductivity in the Time-Reversal Symmetry Breaking Bi/Ni Bilayer System.

Epitaxial bilayer films of Bi(110) and Ni host a time-reversal symmetry breaking superconducting order with an unexpectedly high transition temperature T_{c}=4.1 K. Using time-domain THz spectroscopy, we measure the low energy electrodynamic response of a Bi/Ni bilayer thin film from 0.2 to 2 THz as a function of temperature and magnetic field. We analyze the data in the context of a Bardeen-Cooper-Schrieffer-like superconductor with a finite normal-state scattering rate. In a zero magnetic field, all states in the film become fully gapped, providing important constraints into possible pairing symmetries. Our data appear to rule out the odd-frequency pairing that is natural for many ferromagnetic-superconductor interfaces. By analyzing the magnetic field-dependent response in terms of a pair-breaking parameter, we determine that superconductivity develops over the entire bilayer sample which may point to the p-wave like nature of unconventional superconductivity.