Research in revealing the effects on Cuscuta chinensis to diarrhea type irritable bowel syndrome based on network pharmacology and molecular docking potential mechanism.

To explore the potential mechanism in Cuscuta sinensis on diarrhea-type irritable bowel syndrome using network pharmacology and molecular docking techniques. First, the active components and related targets of Cuscuta were found setting oral utilization >30% and drug-like properties greater than or equal to 0.18 as filter information from TCMSP database. The targets of diarrheal irritable bowel syndrome were compiled by searching DrugBank, GeneCards, OMIM, PharmGkb, and TTD databases. The intersections of drugs and targets related to the disease were taken for gene ontology enrichment and Kyoto encyclopedia of genes and genomes enrichment analyses, to elucidate the potential molecular mechanisms and pathway information of Cuscuta sinensis for the treatment of diarrheal irritable bowel syndrome. The protein-protein interaction network was constructed by using the STRING database and visualized with Cytoscape_v3.10.0 software to find the protein-protein interaction network core At last, molecular docking was performed to validate the combination of active compounds with the core target. The target information of Cuscuta and diarrhea-type irritable bowel syndrome was compiled, which can be resulted in 11 active compounds such as quercetin, kaempferol, isorhamnetin, ß-sitosterol, and another 17 core targets such as TP53, IL6, AKT1, IL1B, TNF, EGFR, etc, whose Kyoto encyclopedia of genes and genomes was enriched in the pathways of lipids and atherosclerosis, chemical carcinogenesis-receptor activation, PI3K-Akt signaling pathway, and fluid shear stress and atherosclerosis, etc. Docking demonstrated that the core targets and the active compounds were able to be better combined. Cuscuta chinensis may exert preventive effects on diarrhea-type irritable bowel syndrome by reducing intestinal inflammation, protecting intestinal mucosa, and playing an important role in antioxidant response through multi-targets and multi-pathways.

Study on the transformation of nitrate nitrogen by manganese-catalyzed iron-carbon micro-electrolysis and microbial coupling.

Nitrate-nitrogen pertains to the nitrogen component of the overall nitrate present in a given sample in order to reduce nitrate nitrogen pollution in water, nitrate nitrogen removal methods based on iron-carbon micro-electrolysis have become a key research focus. The process and mechanism of nitrate nitrogen removal by microbial coupling was comprehensively explored in a novel iron-carbon micro-electrolysis (ICME) system. In order to establish the transformation pathway of nitrate nitrogen in water, the transformation paths of nitrate nitrogen in water before and after coupling microorganisms in three groups of continuous flow reaction devices, namely sponge iron (s-Fe0), sponge iron + biochar (s-Fe0/BC) and sponge iron + biochar + manganese sand (s-Fe0/BC/MS), were studied. The morphology and composition changes of sponge iron were analyzed by means of characterization, and the microbial population changes in the three groups were analyzed by high-throughput sequencing. Results showed that the nitrate conversion rate in the s-Fe0, s-Fe0/BC and s-Fe0/BC/MS systems reached 99.48%, 99.57% and 99.36%, respectively, with corresponding ammonia nitrogen generation, rates of 3.77%, 9.34% and 11.24% and nitrogen generation rates of 95.71%, 90.23% and 88.12%. Scanning electron microscopy imaging showed that in the s-Fe0/BC and s-Fe0/BC/MS systems the surface of sponge iron was highly corroded, with granular substances in the corrosion product clusters. X-ray photoelectron spectroscopy analysis found that the relative contents of Fe2O3 in the surface oxides of sponge iron after microbial coupling were 38.02% and 71.27% in the s-Fe0/BC and s-Fe0/BC/MS systems, while the relative Fe3O4 contents were 61.98% and 28.72%, respectively. Microbial high-throughput sequencing analysis revealed that the Chao and Ace index values in the s-Fe0 system were 871.89 and 880.78, while in the s-Fe0/BC system they were 1012.05 and 1017.29, and in the s-Fe0/BC/MS system were 1241.09 and 1198.29, respectively. The relative proportion of Thauera in the s-Fe0, s-Fe0/BC, and s-Fe0/BC/MS systems was 16.76%,14.25% and 10.01%, while the proportion of Acetoanaerobium was 15.36%, 13.27% and 11.11%, and the proportion of Chloroflexi was 0%, 1.11% and 2.18%, respectively. Furthermore, FAPROTAX function annotation found that the expression levels of chemoheterotrophs in the s-Fe0, s-Fe0/BC and s-Fe0/BC/MS systems were 43 316 OTU, 37 289 OTU and 34 205 OTU, while nitrate respiration expression levels were 16 230 OTU, 15 483 OTU and 9149 OTU, with nitrogen respiration expression levels of 16 328 OTU, 15 493 OTU and 9154 OTU, respectively. These findings suggest that nitrate is converted into nitrogen gas and ammonia nitrogen through the actions of the coupled system of sponge iron/biochar/manganese sand and microorganisms. The catalytic effect of MnO2 promotes the conversion of Fe2+ to Fe3+, generating more electrons, allowing denitrifying bacteria to reduce more nitrate nitrogen, effectively coupling the manganese-catalyzed ICME reaction and microbial denitrification. The micro-electrolysis system and the addition of manganese sand enhanced biodiversity within the s-Fe0/BC/MS system. The heterotrophic bacteria Thauera and Acetoanaerobium were the dominant microorganisms in all three systems, although the micro-electrolysis system with added manganese sand significantly reduced the proportion of facultative bacteria Thauera and Acetoanaerobium and promoted the growth of autotrophic Chloroflexi bacteria. The ecological functions of the three systems were mainly nitrate respiration and nitrogen respiration. By comparing the expression levels of nitrate respiration and nitrogen respiration in s-Fe0/BC and s-Fe0/BC/MS systems, it can be seen that the addition of manganese sand reduced microbial activity.

LncRNA STAT3-AS regulates endometrial receptivity via the STAT3 signaling pathway.

Endometrial receptivity is critical for the successful establishment of pregnancy in ruminants. Interferon tau (IFNT) plays a key role in promoting embryo attachment by activating the Janus kinase/signal transducer and activator of transcription pathway, which induces the expression of a series of interferon-stimulated genes (ISGs). In our previous study, sequencing analysis of goat endometrial epithelial cells (gEECs) treated with 20 ng/mL IFNT revealed a differentially expressed long non-coding RNA located on the STAT3 antisense chain, which we designated STAT3-AS. The aim of this study was to investigate the role and mechanism of STAT3-AS in establishing endometrial receptivity in goats. The results showed that STAT3-AS was expressed in both the nucleus and cytoplasm of gEECs, and its expression increased significantly in the uterus on day 15 of pregnancy. STAT3-AS expression was upregulated in gEECs treated with IFNT alone or in combination with progesterone and estradiol. Knockdown of STAT3-AS using specific short interfering RNA significantly inhibited the expression of classical ISGs such as interferon-stimulated gene 15 and 2',5'-oligodenylate synthetase 2, as well as uterine endometrial receptivity-related genes including homeobox gene A11, integrin beta 3, and vascular endothelial growth factor. Moreover, gEEC proliferation and the STAT3 pathway were suppressed in the absence of STAT3-AS. However, pretreatment with the STAT3 activator RO8191 restored the effect of silencing STAT3-AS on endometrial receptivity. Overall, these results suggest that STAT3-AS is an important regulator of endometrial receptivity in goats and that it regulates endometrial receptivity through the STAT3 pathway.

Bowen's disease of the vulva and perianal area in a SLE patient successfully treated with Photodynamic therapy: A case report.

Photodynamic therapy (PDT) has emerged as a non-invasive treatment modality for superficial skin cancers. It has the advantage of greater tolerance and providing better cosmetic outcomes than conventional treatment methods. Because of the rarity of extensive Bowen's disease located in the genital area, evidence of efficacy for therapies is mainly based on case reports and clinical experience. This report presents a case of a 32-year-old female with Bowen's disease of the vulva and perianal area with systemic lupus erythematosus successfully treated by 5-aminolaevulinic acid PDT. There was no evidence of recurrence after five-years of follow-up.

HMGB1 Signaling-Mediated Tumor Immunity in Cancer Progress.

Tumor immunity is a cycle that begins with the release of antigens from tumor cells and ends with the destruction of tumor cells. High mobility group box 1 (HMGB1) is a nonhistone protein widely present in the nucleus of mammalian cells and can be released by immune cells or tumor cells. As a proinflammatory mediator or alarm protein, the activity and function of HMGB1 are determined by the environment, binding receptors, redox status and posttranslational modifications (PTMs), and HMGB1 plays a key role in inflammation and tumor immune processes. In this review, we summarize in detail the current studies on the dual role of HMGB1 in tumor immunity, focusing mainly on immunosuppressive effects, such as regulatory T cells (Tregs), myeloid-derived suppressor cells (MDSCs) and tumor-associated macrophages (TAMs), as well as antitumor immunoenhancement effects, such as immunogenic cell death (ICD). Finally, we discuss the potential and challenges of HMGB1 in antitumor immunotherapy.

IDR-induced CAR condensation improves the cytotoxicity of CAR-Ts against low-antigen cancers.

Chimeric antigen receptor (CAR)-T cell-based therapies demonstrate remarkable efficacy for the treatment of otherwise intractable cancers, particularly B-cell malignancies. However, existing FDA-approved CAR-Ts are limited by low antigen sensitivity, rendering their insufficient targeting to low antigen-expressing cancers. To improve the antigen sensitivity of CAR-Ts, we engineered CARs targeting CD19, CD22, and HER2 by including intrinsically disordered regions (IDRs) that promote signaling condensation. The "IDR CARs" triggered enhanced membrane-proximal signaling in the CAR-T synapse, which led to an increased release of cytotoxic factors, a higher killing activity towards low antigen-expressing cancer cells in vitro, and an improved anti-tumor efficacy in vivo. No elevated tonic signaling was observed in IDR CAR-Ts. Together, we demonstrated IDRs as a new tool set to enhance CAR-T cytotoxicity and to broaden CAR-T's application to low antigen-expressing cancers.

Low-Velocity-Favored Transition Radiation.

When a charged particle penetrates through an optical interface, photon emissions emerge-a phenomenon known as transition radiation. Being paramount to fundamental physics, transition radiation has enabled many applications from high-energy particle identification to novel light sources. A rule of thumb in transition radiation is that the radiation intensity generally decreases with the decrease of particle velocity v; as a result, low-energy particles are not favored in practice. Here, we find that there exist situations where transition radiation from particles with extremely low velocities (e.g., v/c<10^{-3}) exhibits comparable intensity as that from high-energy particles (e.g., v/c=0.999), where c is the light speed in free space. The comparable radiation intensity implies an extremely high photon extraction efficiency from low-energy particles, up to 8 orders of magnitude larger than that from high-energy particles. This exotic phenomenon of low-velocity-favored transition radiation originates from the interference of the excited Ferrell-Berreman modes in an ultrathin epsilon-near-zero slab. Our findings may provide a promising route toward the design of integrated light sources based on low-energy electrons and specialized detectors for beyond-standard-model particles.

Interfacial Cherenkov radiation from ultralow-energy electrons.

Cherenkov radiation occurs only when a charged particle moves with a velocity exceeding the phase velocity of light in that matter. This radiation mechanism creates directional light emission at a wide range of frequencies and could facilitate the development of on-chip light sources except for the hard-to-satisfy requirement for high-energy particles. Creating Cherenkov radiation from low-energy electrons that has no momentum mismatch with light in free space is still a long-standing challenge. Here, we report a mechanism to overcome this challenge by exploiting a combined effect of interfacial Cherenkov radiation and umklapp scattering, namely the constructive interference of light emission from sequential particle-interface interactions with specially designed (umklapp) momentum-shifts. We find that this combined effect is able to create the interfacial Cherenkov radiation from ultralow-energy electrons, with kinetic energies down to the electron-volt scale. Due to the umklapp scattering for the excited high-momentum Bloch modes, the resulting interfacial Cherenkov radiation is uniquely featured with spatially separated apexes for its wave cone and group cone.

Free-electron Brewster-transition radiation.

We reveal a mechanism to enhance particle-matter interactions by exploiting the pseudo-Brewster effect of gain materials, presenting an enhancement of at least four orders of magnitude for light emission. This mechanism is enabled by the emergence of an unprecedented phase diagram that maps all phenomena of free-electron transition radiation into three distinct phases in a gain-thickness parameter space, namely, the conventional, intermediate, and Brewster phases, when an electron penetrates a dielectric slab with a modest gain and a finite thickness. Essentially, our revealed mechanism corresponds to the free-electron transition radiation in the Brewster phase, which also features ultrahigh directionality, always at the Brewster angle, regardless of the electron velocity. Counterintuitively, we find that the intensity of this free-electron Brewster-transition radiation is insensitive to the Fabry-Pérot resonance condition and, thus, the variation of slab thickness, and moreover, a weaker gain could lead to a stronger enhancement for light emission.

Emergent Peptides of the Antifibrotic Arsenal: Taking Aim at Myofibroblast Promoting Pathways.

Myofibroblasts are the principal effector cells driving fibrosis, and their accumulation in tissues is a fundamental feature of fibrosis. Essential pathways have been identified as being central to promoting myofibroblast differentiation, revealing multiple targets for intervention. Compared with large proteins and antibodies, peptide-based therapies have transpired to serve as biocompatible and cost-effective solutions to exert biomimicry, agonistic, and antagonistic activities with a high degree of targeting specificity and selectivity. In this review, we summarize emergent antifibrotic peptides and their utilization for the targeted prevention of myofibroblasts. We then highlight recent studies on peptide inhibitors of upstream pathogenic processes that drive the formation of profibrotic cell phenotypes. We also briefly discuss peptides from non-mammalian origins that show promise as antifibrotic therapeutics. Finally, we discuss the future perspectives of peptide design and development in targeting myofibroblasts to mitigate fibrosis.

Knockdown of HMGB1 inhibits the crosstalk between oral squamous cell carcinoma cells and tumor-associated macrophages.

Tumor-associated macrophages (TAMs), the major component of the tumor microenvironment (TME), play distinctly different roles in different tumors. High mobility group box 1 (HMGB1), a nonhistone protein in the nucleus, can perform functions during inflammation and cancers. However, the role of HMGB1 in the crosstalk between oral squamous cell carcinoma (OSCC) cells and TAMs remains unclear. Here, we established a coculture system of TAMs and OSCC cells to explore the bidirectional effect and potential mechanism of HMGB1 in OSCC cell-TAM interactions. Our results showed that HMGB1 was significantly upregulated in OSCC tissues and positively associated with tumor progression, immune cell infiltration and macrophage polarization. Then, knocking down HMGB1 in OSCC cells inhibited the recruitment and polarization of cocultured TAMs. Moreover, the knockdown of HMGB1 in macrophages not only suppressed polarization, but also inhibited cocultured OSCC cell proliferation, migration and invasion in vitro and in vivo. Mechanistically, macrophages secreted higher levels of HMGB1 than OSCC cells, and dampening endogenous HMGB1 reduced HMGB1 secretion. Both OSCC cell-generated and macrophage-endogenous HMGB1 may regulate TAM polarization by promoting receptor TLR4 expression and NF-κB/p65 activation and enhancing IL-10/TGF-ß expression. HMGB1 in OSCC cells may regulate macrophage recruitment via IL-6/STAT3. In addition, TAM-derived HMGB1 may affect aggressive phenotypes of cocultured OSCC cells by regulating the immunosuppressive microenvironment through the IL-6/STAT3/PD-L1 and IL-6/NF-κB/MMP-9 pathways. In conclusion, HMGB1 may regulate the crosstalk between OSCC cells and TAMs, including modulating macrophage polarization and attraction, enhancing cytokine secretion, and remodeling and creating an immunosuppressive TME to further affect OSCC progression.

Effect of anesthesia assistance on the detection rate of precancerous lesions and early esophageal squamous cell cancer in esophagogastroduodenoscopy screening: A retrospective study based on propensity score matching.

Background: Esophagogastroduodenoscopy (EGD) screening is vital for the early diagnosis of esophageal squamous cell cancer (ESCC). However, improvement in the detection rate of precancerous lesions and early ESCC with anesthesia assistance (AA) has not yet been investigated. This retrospective study aimed to evaluate the effect of AA on the detection rate of precancerous lesions and early ESCC in patients undergoing EGD screening and identify risk factors affecting the detection rate. Methods: We reviewed patients' electronic medical records who underwent EGD screening between May 2019 and August 2020. Patients were divided into two groups based on whether they received AA: those in Group A underwent EGD screening with AA, and patients in Group O underwent EGD screening without AA. Propensity score matching (PSM) was used to account for differences in baseline characteristics. Detection rates of precancerous lesions and early ESCC were compared between the two groups following PSM. Binary logistic regression was used to identify risk factors affecting the detection rate. Results: The final analysis included 21,835 patients (Group A = 13,319, Group O = 8,516) from 28,985 patients who underwent EGD screening during the study period. Following PSM, 6009 patients remained in each group for analysis. There was no significant difference in the detection rate of precancerous lesions and early ESCC between Groups A and O (1.1% vs. 0.8%, p > 0.05). Binary logistic regression showed that age (50-59 years, 60-69 years and 70-79 years), higher endoscopist seniority, high-definition (HD) endoscopy, narrow-band imaging (NBI), and number of endoscopic images were all independent risk factors that affected the detection rate of precancerous lesions and early ESCC. Conclusion: There was no statistically significant difference in the detection rate of precancerous lesions and early ESCC between patients who underwent EGD screening with and without AA. All independent risk factors that affected the detection rate of precancerous lesions and early ESCC included the following: age (50-59 years, 60-69 years and 70-79 years), higher endoscopist seniority, HD endoscopy, NBI, and number of endoscopic images. Endoscopists should consider all these factors as much as possible when performing EGD screening.

Electrosynthesis of polymer-grade ethylene via acetylene semihydrogenation over undercoordinated Cu nanodots.

The removal of acetylene impurities remains important yet challenging to the ethylene downstream industry. Current thermocatalytic semihydrogenation processes require high temperature and excess hydrogen to guarantee complete acetylene conversion. For this reason, renewable electricity-based electrocatalytic semihydrogenation of acetylene over Cu-based catalysts is an attractive route compared to the energy-intensive thermocatalytic processes. However, active Cu electrocatalysts still face competition from side reactions and often require high overpotentials. Here, we present an undercoordinated Cu nanodots catalyst with an onset potential of -0.15 V versus reversible hydrogen electrode that can exclusively convert C2H2 to C2H4 with a maximum Faradaic efficiency of ~95.9% and high intrinsic activity in excess of -450 mA cm-2 under pure C2H2 flow. Subsequently, we successfully demonstrate simulated crude ethylene purification, continuously producing polymer-grade C2H4 with <1 ppm C2H2 for 130 h at a space velocity of 1.35 × 105 ml gcat-1 h-1. Theoretical calculations and in situ spectroscopies reveal a lower energy barrier for acetylene semihydrogenation over undercoordinated Cu sites than nondefective Cu surface, resulting in the excellent C2H2-to-C2H4 catalytic activity of Cu nanodots.

USP18 promotes endometrial receptivity via the JAK/STAT1 and the ISGylation pathway.

Interferon-tau (IFNT), a pregnancy recognition signal in ruminants, promotes the establishment of endometrial receptivity by inducing the expression of interferon-stimulated genes (ISGs) via the Janus kinase/signal transducer and activator of transcription (JAK/STATs) signaling pathway. However, the precise mechanisms remain largely unknown. Ubiquitin-specific protease 18 (USP18) acts specifically on the ISGylation modification system to exert deubiquitination and participates in the regulation of the type I IFN signaling pathway. The purpose of this study was to determine the role and mechanism of USP18 on endometrial receptivity in goat. USP18 was mainly localized in the uterine luminal and glandular epithelium, and its expression levels were significantly increased from days 5-18 of early pregnancy. Progesterone (P4), estradiol (E2), and IFNT significantly stimulated USP18 expression in goat endometrial epithelial cells (gEECs) cultured in vitro. Meanwhile, the markers of endometrial receptivity HOXA11, ITGB1, ITGB3, and ITGB5 were significantly upregulated after USP18 overexpression in gEECs. However, USP18 interference significantly inhibited the expression of HOXA10, ITGB1, ITGB3, and ITGB5 in gEECs. In addition, both the phosphorylation levels of STAT1 and the expression of ISGylation-modified proteins were significantly increased after USP18 silencing in gEECs. Furthermore, pretreatment with the STAT1 inhibitor Fludara markedly restored the effect of USP18 interference in gEECs. In summary, USP18 may play an important role in promoting goat endometrial receptivity by regulating the JAK/STAT1 pathway and ISGylation.

Hypoxia-mediated chemotaxis and residence of macrophage in decidua by secreting VEGFA and CCL2 during normal pregnancy.

In brief: Hypoxia is vital for the establishment of the maternal-fetal interface during early pregnancy. This study shows that decidual macrophages (dMφ) could be recruited and reside in decidua under the regulation of hypoxia/VEGFA-CCL2 axis. Abstract: Infiltration and residence of decidual macrophages (dMφ) are of great significance to pregnancy maintenance for their role in angiogenesis, placental development, and inducing immune tolerance. Besides, hypoxia has now been acknowledged as an important biological event at maternal-fetal interface in the first trimester. However, whether and how hypoxia regulates biofunctions of dMφ remain elusive. Herein, we observed increased expression of C-C motif chemokine ligand 2 (CCL2) and residence of macrophages in decidua compared to secretory-phase endometrium. Moreover, hypoxia treatment on stromal cells improved the migration and adhesion of dMφ. Mechanistically, these effects might be mediated by upregulated CCL2 and adhesion molecules (especially ICAM2 and ICAM5) on stromal cells in the presence of endogenous vascular endothelial growth factor-A (VEGFA) in hypoxia. These findings were also verified by recombinant VEGFA and indirect coculture, indicating that the interaction between stromal cells and dMφ in hypoxia condition may facilitate dMφ recruitment and residence. In conclusion, VEGFA derived from a hypoxic environment may manipulate CCL2/CCR2 and adhesion molecules to enhance the interactions between dMφ and stromal cells and thus contribute to the enrichment of macrophages in decidua early during normal pregnancy.

Application of simultaneous ultrasonic curing on pork (Longissimus dorsi): Mass transport of NaCl, physical characteristics, and microstructure.

This study aimed to investigate the effect of ultrasound curing with various working modes and frequency combinations, including mono-, dual- and tri-frequency, on the content of NaCl and tenderness of pork loins (Longissimus dorsi). The physical qualities, myoglobin, moisture migration, distribution, and microstructure of pork were also evaluated. The results displayed that the NaCl content of samples cured by simultaneous ultrasound (100 W/L) working mode with a frequency combination of 20, 40, and 60 kHz was higher than that of other ultrasound working modes. The effect of ultrasonic brining was significantly better than the static curing when the saline solution was >35 mL. In addition, the samples cured by simultaneous ultrasound had better physical qualities, including more pickling absorptivity, less cooking loss, and lower hardness, tenderness, and chewiness value. The intensity of lightness was reduced, although redness and yellowness remained unaltered compared to static curing. The myoglobin content decreased drastically without changing the oxygenation level, and the relaxation time of T2b and T21 was delayed. The microstructure indicated that the ultrasonic treatment could promote changes in meat texture. Overall, the simultaneous ultrasound at various frequencies could efficiently accelerate NaCl penetration and improve pork quality.

PROTAC Degrader of Estrogen Receptor α Targeting DNA-Binding Domain in Breast Cancer.

PROteolysis-TArgeting Chimeras (PROTACs) are a powerful class of drugs that selectively degrade the proteins of interest (POIs) through cellular ubiquitination mechanisms. Estrogen receptor α (ERα) plays a vital role in the pathogenesis and treatment of breast cancer. In this work, the DNA-binding domain (DBD) of ERα was selected as the target to avoid drug resistance caused by the ligand-binding domain (LBD) of ERα. The estrogen response element (ERE), a natural DNA sequence binding with DBD of ERα, was chosen as a recognized unit of PROTAC. Therefore, we designed a nucleic acid-conjugated PROTAC, ERE-PROTAC, via a click reaction, in which the ERE sequence recruits ERα and the typical small molecule VH032 recruits the von Hippel-Lindau (VHL) E3 ligase. The proposed ERE-PROTAC showed to efficiently and reversibly degrade ERα in different breast cancer cells by targeting the DBD, indicating its potential to overcome the current resistance caused by LBD mutations.

Low-density lipoprotein receptor deficiency reduced bone mass in mice via the c-fos/NFATc1 pathway.

AIM: The low-density lipoprotein receptor (LDLR) plays a crucial role in regulating lipid metabolism. However, whether LDLR deficiency affects bone mass and morphology remains controversial. This study aimed to analyze the bone phenotypes of LDLR knockout (LDLR-/-) mice. MAIN METHODS: Eight-week-old LDLR-/- and wild-type (WT) mice were subjected to microcomputed tomography to detect bone phenotypes. Enzyme-linked immunosorbent assay kits were used to detect the serum estrogen levels and matrix metalloproteinase 9 (MMP-9) levels in tissue homogenates. Von Kossa, toluidine blue, tartrate-resistant acid phosphatase (TRAP) staining, and calcein labeling were performed to explore bone turnover parameters. In vitro, osteoclastogenesis was induced in bone marrow cells from LDLR-/- mice and WT mice in the presence or absence of 17ß-estradiol. The microphotographs and number of osteoclasts were validated using TRAP staining. Relative gene expression during osteoclast differentiation and maturation was determined by quantitative real-time polymerase chain reaction. KEY FINDINGS: LDLR deficiency results in reduced bone mineral density of the tibial cancellous bone, indicating bone loss to some extent in LDLR-/- mice. LDLR deficiency significantly increased the number of osteoclasts, but not osteoblasts. In vitro, bone marrow cells from LDLR-/- mice displayed enhanced osteoclastic potential along with increased expression of TRAP, cathepsin K, nuclear factor of activated T-cells 1 (NFATc1), c-fos, and MMP-9 and inhibited dendritic cell-specific transmembrane protein expression. Moreover, 17ß-estradiol treatment can inhibit osteoclastogenesis in vitro. SIGNIFICANCE: Our data demonstrated that LDLR deficiency promoted osteoclastogenesis by upregulating c-fos and NFATc1 expression, reducing cancellous bone mass in LDLR-/- mice.

Biallelic mutations of TTC12 and TTC21B were identified in Chinese patients with multisystem ciliopathy syndromes.

BACKGROUND: Abnormalities in cilia ultrastructure and function lead to a range of human phenotypes termed ciliopathies. Many tetratricopeptide repeat domain (TTC) family members have been reported to play critical roles in cilium organization and function. RESULTS: Here, we describe five unrelated family trios with multisystem ciliopathy syndromes, including situs abnormality, complex congenital heart disease, nephronophthisis or neonatal cholestasis. Through whole-exome sequencing and Sanger sequencing confirmation, we identified compound heterozygous mutations of TTC12 and TTC21B in six affected individuals of Chinese origin. These nonsynonymous mutations affected highly conserved residues and were consistently predicted to be pathogenic. Furthermore, ex vivo cDNA amplification demonstrated that homozygous c.1464 + 2 T > C of TTC12 would cause a whole exon 16 skipping. Both mRNA and protein levels of TTC12 were significantly downregulated in the cells derived from the patient carrying TTC12 mutation c.1464 + 2 T > C by real-time qPCR and immunofluorescence assays when compared with two healthy controls. Transmission electron microscopy analysis further identified ultrastructural defects of the inner dynein arms in this patient. Finally, the effect of TTC12 deficiency on cardiac LR patterning was recapitulated by employing a morpholino-mediated knockdown of ttc12 in zebrafish. CONCLUSIONS: To the best of our knowledge, this is the first study reporting the association between TTC12 variants and ciliopathies in a Chinese population. In addition to nephronophthisis and laterality defects, our findings demonstrated that TTC21B should also be considered a candidate gene for biliary ciliopathy, such as TTC26, which further expands the phenotypic spectrum of TTC21B deficiency in humans.