Paris polyphylla ethanol extract and polyphyllin I ameliorate adenomyosis by inhibiting epithelial-mesenchymal transition.

BACKGROUND: The active ingredients of the Chinese medical herb Paris polyphylla, P. polyphylla ethanol extract (PPE) and polyphyllin I (PPI), potentially inhibit epithelial-mesenchymal transition (EMT) in tumors. However, the roles of these ingredients in inhibiting EMT in adenomyosis (AM) remain to be explored. PURPOSE: The primary goal of the study was to uncover the underlying molecular processes through which PPE and PPI suppress EMT in AM, alongside assessing the safety profiles of these substances. METHODS: To assess the suppressive impact of PPE on adenomyosis-derived cells (AMDCs), we employed Transwell and wound healing assays. The polyphyllins (PPI, PPII, PPVII) contained in PPE were characterized using high-performance liquid chromatography (HPLC). Then, bioinformatics techniques were performed to pinpoint potential PPI targets that could be effective in treating AM. Immunoblotting was used to verify the key proteins and pathways identified via bioinformatics. Furthermore, we examined the efficacy of PPE and PPI in treating Institute of Cancer Research (ICR) mice with AM by observing the morphological and pathological features of the uterus and performing immunohistochemistry. In addition, we assessed safety by evaluating liver, kidney and spleen pathologic features and serum test results. RESULTS: Three major polyphyllins of PPE were revealed by HPLC, and PPI had the highest concentration. In vitro experiments indicated that PPE and PPI effectively prevent AMDCs invasion and migration. Bioinformatics revealed that the primary targets E-cadherin, N-cadherin and TGFß1, as well as the EMT biological process, were enriched in PPI-treated AM. Immunoblotting assays corroborated the hypothesis that PPE and PPI suppress the TGFß1/Smad2/3 pathway in AMDCs to prevent EMT from progressing. Additionally, in vivo studies showed that PPE (3 mg/kg and 6 mg/kg) and PPI (3 mg/kg and 6 mg/kg), successfully suppressed the EMT process through targeting the TGFß1/Smad2/3 signaling pathway. Besides, it was observed that lower doses of PPE (3 mg/kg) and PPI (3 mg/kg) exerted minimal effects on the liver, kidneys, and spleen. CONCLUSIONS: PPE and PPI efficiently impede the development of EMT by inhibiting the TGFß1/Smad2/3 pathway, revealing an alternative pathway for the pharmacological treatment of AM.

[Developments in Research on the Relationship Between Porphyromonas gingivalis and Non-Oral Diseases].

Porphyromonas gingivalis ( P. gingivalis) is a common periodontal pathogen. Recently, there has been increasing evidence suggesting that P. gingivalis is not only a common pathogen in the oral cavity, but is also closely associated with non-oral diseases, including inflammatory bowel disease, cancer, cardiovascular diseases, Alzheimer's disease, rheumatoid arthritis, diabetes mellitus, premature birth and non-alcoholic hepatitis, etc. Herein, we reviewed the developments in recent years in research on the relationship between P. gingivalis, a periodontal pathogen, and non-oral diseases, which will help determine whether P. gingivalis could be used as an auxiliary diagnostic biomarker or a potential therapeutic target for these non-oral diseases, thus contributing to the development of treatment strategies for the relevant diseases.

Sulfated alginate oligosaccharide exerts antitumor activity and autophagy induction by inactivating MEK1/ERK/mTOR signaling in a KSR1-dependent manner in osteosarcoma.

Alginate oligosaccharide (AOS) has the function to inhibit tumor progression and the sulfated modification can enhance the antitumor activity. To date, the function and mechanism of sulfated AOS (AOS-SO4) in tumors remain largely elusive. We prepared AOS by the enzymatic degradation of alginate, collected AOS-SO4 by sulfating following the canonical procedure. Using these materials, in vitro assays showed that both AOS and AOS-SO4 elicited antitumor effects in osteosarcoma cells. Sulfated modification significantly enhanced the antitumor activity. In addition, AOS-SO4 had obvious effects on cell cycle arrest, apoptosis, and autophagy induction in vitro and in vivo. Mechanistically, we observed that AOS-SO4 treatment triggered proapoptotic autophagy by inhibiting MEK1/ERK/mTOR signaling. The ERK activator reversed AOS-SO4-induced autophagy. More importantly, we found that KSR1 interacted with MEK1 and functioned as a positive regulator of MEK1 protein in osteosarcoma cells. High KSR1 expression was significantly associated with poor survival in osteosarcoma patients. Together, these results suggest that AOS-SO4 has a better antitumor effect in osteosarcoma by inhibiting MEK1/ERK/mTOR signaling, which is KSR1-dependent; thus, AOS-SO4 can be a new potential therapeutic candidate for the treatment of osteosarcoma.

Genetically Encoded Sensor Enables Endogenous RNA Imaging with Conformation-Switching Induced Fluorogenic Proteins.

Genetically encoded molecular tools are crucial for live cell RNA imaging, and few are available for endogenous RNA imaging. We develop a new genetically encoded sensor using conformation switching RNA induced fluorogenic proteins that enable multicolor and signal-amplified imaging of endogenous RNAs. The sensor system is designed with an RNA sensing module and a degron-fused fluorescent protein reporter. Target RNA induces conformation switching of the RNA sensing module to form RNA aptamers that stabilize the degron-fused protein for fluorogenic imaging. This sensor is demonstrated for high-contrast imaging of survivin mRNA abundance and dynamics in live cells. Moreover, the sensor system is extended to a multicolor palette by screening fluorogenic proteins of distinct colors, and engineered into a signal amplifier using the split fluorescent protein design. The sensor is further exploited for imaging lncRNA MALAT-1 and its translocation dynamics during mitosis. Our sensor system can afford a valuable platform for RNA imaging in biomedical research and clinical theranostics.

Siderotic cataract with no signs of intraocular foreign body.

BACKGROUND: Ocular siderosis is a clinical condition induced by deposition of an iron-containing intraocular foreign body. We report a unique case of histopathologically proven lens siderosis in a young woman with a preceding history of trauma but no signs of retained intraocular foreign body. CASE PRESENTATION: A 32-year-old woman presented with an opacified lens showing brownish deposits on the anterior capsule and underwent cataract surgery. Preoperative ophthalmic examination did not show any retained intraocular foreign body. Histopathologic staining of the anterior capsule confirmed the presence of iron deposits and macrophages. Electroretinography examination performed in the postoperative period showed the changes characteristic of retinal degeneration in ocular siderosis. CONCLUSION: This case illustrates the importance of close monitoring of patients with a history of trauma or previous penetrating injury to the eye, even if there is no intraocular foreign body, because they might develop ocular siderosis at a later stage. This case report underscores the importance of electroretinography and histopathologic analysis, in addition to ophthalmic examination, in the diagnosis of ocular siderosis.

Diagnosis of pupillary block glaucoma after removal of congenital cataracts with intraoperative ultrasound biomicroscopy: a case report.

BACKGROUND: Aphakic glaucoma is a common complication after congenital cataract extraction, especially in those who have surgery during infancy. This case report describes a case of bilateral pupillary block glaucoma diagnosed with intraoperative ultrasound biomicroscopy (UBM) after removal of congenital cataract. CASE PRESENTATION: We present a case report of a 9-month-old infant with bilateral corneal enlargement and ocular hypertension after uneventful removal of congenital cataracts. Initial and follow-up examination findings were reviewed. The infant was suspected to have developmental glaucoma and schemed to have bilateral trabeculotomy until pupillary obstruction by vitreous herniation and angle closure with iris bombé were detected by intraoperative UBM. Anterior vitrectomy and goniosynechialysis were then performed as treatment. CONCLUSION: Pupillary block glaucoma is a rare type of infantile aphakic glaucoma. Application of intraoperative UBM can assist in the differential diagnosis of aphakic glaucoma in infants.

Elevated TGF-ß2 level in aqueous humor of cataract patients with high myopia: Potential risk factor for capsule contraction syndrome.

PURPOSE: To evaluate the level of transforming growth factor-ß2 (TGF-ß2) in the aqueous humor of highly myopic cataract patients and its correlation with capsule contraction syndrome. SETTING: Eye and ENT Hospital of Fudan University, Shanghai, China. DESIGN: Prospective comparative case series. METHODS: The highly myopic cataract patients were divided into the following 2 groups according to the Lens Opacity Classification System III: nuclear color (NC) 2 to 3 and NC 5 to 6. Aqueous humor TGF-ß2 concentrations were assayed in the highly myopic cataract and age-related cataract groups. The TGF-ß2, TGF-ßRII (the type II receptor for TGF-ß2), and α-smooth muscle actin (α-SMA) expressions in lens epithelial cells (LECs) were detected by real-time polymerase chain reaction and immunofluorescent staining. RESULTS: The study comprised 40 highly myopic cataract patients (40 eyes) and 20 patients (20 eyes) with age-related cataract as the control group. Compared with the control group, the highly myopic cataract group had significantly higher TGF-ß2 concentration in the aqueous humor and increased TGF-ßRII expression in LECs, especially in NC 5 to 6 cases. Expression of α-SMA was barely detectable in both groups. CONCLUSION: In highly myopic cataract patients, especially those with dark nuclei, elevated aqueous humor TGF-ß2 levels and the upregulated TGF-ßRII expression in LECs might contribute to the pathogenesis of capsule contraction syndrome through transdifferentiation of LECs into myofibroblasts. FINANCIAL DISCLOSURE: None of the authors has a financial or proprietary interest in any material or method mentioned.

Molecular Inflammation in the Contralateral Eye After Cataract Surgery in the First Eye.

PURPOSE: The purpose of this study was to assess the inflammatory status of the aqueous humor in the fellow eye after uneventful cataract surgery in the first eye. METHODS: At the screening stage, aqueous humor samples from 15 first-eye and 15 second-eye cataract patients were collected just before cataract surgery and assayed using human cytokine antibody array. Screened cytokines were then verified using a suspension array system with aqueous humor samples obtained from 35 first-eye and 36 second-eye cataract patients. RESULTS: The cytokine antibody array revealed that interleukin-1 receptor antagonist (Il-1ra) and macrophage inflammatory protein (MIP)-1a and MIP-1b were expressed at high levels in first-eye patients and were lower in second-eye patients, whereas opposite trends were found for monocyte chemoattractant protein 1 (MCP-1) and for regulated on activation, normal T expressed and secreted (RANTES) (all, P < 0.05, Student's t-test). However, only MCP-1 and IL-1ra were significantly different between the two groups after Bonferroni correction (both P < 0.00125). In the replication stage, the suspension cytokine array revealed that only MCP-1 expression was significantly greater in the aqueous humor of second-eye patients than in that of first-eye patients (P = 0.0067, Student's t-test). CONCLUSIONS: This study revealed that expression of MCP-1, a pain-related inflammatory chemokine, was significantly increased in aqueous humor in the contralateral eye after first-eye cataract surgery. This suggests there may be a sympathetic ophthalmic type uveitis in the contralateral eye after first-eye cataract surgery and that may help to explain why second-eye phacoemulsification is often more painful. (ClinicalTrials.gov number, NCT01824927.)

Plasma-enhanced microwave solid-state synthesis of cadmium sulfide: reaction mechanism and optical properties.

CdS synthesis by plasma-enhanced microwave physical vapor transport (PMPVT) has been developed in this work. The photoluminescence (PL), absorbance, Raman spectra and the mechanism of CdS crystal growth have been investigated. Furthermore, plasma-enhanced microwave chemical vapour transport (PMCVT) synthesis of CdS with additional chemical transport agents has been explored. In addition, other II-VI chalcogenides were also synthesized by PMPVT.

αA-crystallin gene CpG islands hypermethylation in nuclear cataract after pars plana vitrectomy.

PURPOSE: To investigate the DNA methylation status of αA-crystallin gene in cataract secondary to pars plana vitrectomy. METHODS: Anterior capsular membranes of 40 eyes of 40 patients with cataract secondary to vitrectomy were collected. Another 20 eyes of 20 patients who received pars plana vitrectomy and phacoemulsification in the primary procedure, were recruited as control. Methylation status of the CpG islands of αA-crystallin gene was analyzed by pyrosequencing. Expression of αA-crystallin was evaluated by real-time polymerase chain reaction and western blot. RESULTS: In the post vitrectomy group, five patients with posterior subcapsular opacity and four patients with cortical opacity were excluded from further analysis. The remaining 31 patients with nuclear cataract were assigned into two groups according to tamponade types: 19 of octafluoropropane (C3F8) and 12 of silicone oil (SiO). The average nuclear color grading was elevated both in C3F8 and SiO groups after vitrectomy. Compared to the control group, hypermethylation of the CpG islands in the αA-crystallin gene promoter was found in both post vitrectomy groups, accompanied by significantly reduced αA-crystallin expression. No statistically significant differences were found between the C3F8 and SiO groups either for DNA methylation status or αA-crystallin expression. CONCLUSIONS: CpG islands hypermethylation of αA-crystallin gene may be involved in nuclear cataract formation after pars plana vitrectomy.

Dual-color labeled anti-mucin 1 antibody for imaging of ovarian cancer: A preliminary animal study.

To investigate the feasibility of the anti-mucin 1 (anti-MUC1/CD227) antibody in the fluorescent imaging of ovarian cancer, the CD227 antibody and a control IgG antibody were labeled with a near-infrared dye [Cy5.5-N-hydroxysuccinimide (NHS)] and a green dye (fluorescein-NHS). In vivo fluorescence images were obtained at 4, 12 and 36 h after injection of the probes into OVCAR3 tumor-bearing mice. The tumor to background ratios were calculated for both probes. Ex vivo fluorescence images were obtained following sacrifice at 36 h. After conjugation to Cy5.5 and fluorescein, the dual-color labeled CD227 probe (Ab-FL-Cy5.5) could be visualized by both green and near-infrared fluorescence. Uptake by the tumors was higher for the Ab-FL-Cy5.5 than for the IgG-Cy5.5 probe. All tumors could be visualized by in vivo imaging with an acceptable tumor to background ratio. Ex vivo studies demonstrated the advantages of using green fluorescence imaging to guide the resection of tumor tissues. These preliminary data indicate that the Ab-FL-Cy5.5 probe is promising for further tumor imaging applications and clinical translation.

Identification of proteins that interact with alpha A-crystallin using a human proteome microarray.

PURPOSE: To identify proteins interacting with alpha A-crystallin (CRYAA) and to investigate the potential role that these protein interactions play in the function of CRYAA using a human proteome (HuProt) microarray. METHODS: The active full-length CRYAA protein corresponding to amino acids 1-173 of CRYAA was recombined. A HuProt microarray composed of 17,225 human full-length proteins with N-terminal glutathione S-transferase (GST) tags was used to identify protein-protein interactions. The probes were considered detectable when the signal to noise ratio (SNR) was over 1.2. The identified proteins were subjected to subsequent bioinformatics analysis using the DAVID database. RESULTS: The HuProt microarray results showed that the signals of 343 proteins were higher in the recombinant CRYAA group than in the control group. The SNR of 127 proteins was ≥ 1.2. The SNR of the following eight proteins was > 3.0: hematopoietic cell-specific Lyn substrate 1 (HCLS1), Kelch domain-containing 6 (KLHDC6), sarcoglycan delta (SGCD), KIAA1706 protein (KIAA1706), RNA guanylyltransferase and 5'-phosphatase (RNGTT), chromosome 10 open reading frame 57 (C10orf57), chromosome 9 open reading frame 52 (C9orf52), and plasminogen activator, urokinase receptor (PLAUR). The bioinformatics analysis revealed 127 proteins associated with phosphoproteins, alternative splicing, acetylation, DNA binding, the nuclear lumen, ribonucleotide binding, the cell cycle, WD40 repeats, protein transport, transcription factor activity, GTP binding, and cellular response to stress. Functional annotation clustering showed that they belong to cell cycle, organelle or nuclear lumen, protein transport, and DNA binding and repair clusters. CRYAA interacted with these proteins to maintain their solubility and decrease the accumulation of denatured target proteins. The protein-protein interactions may help CRYAA carry out multifaceted functions. CONCLUSIONS: One-hundred and twenty-seven of 17,225 human full-length proteins were identified that interact with CRYAA. The advent of microarray analysis enables a better understanding of the functions of CRYAA as a molecular chaperone.

Epigenetic regulation of αA-crystallin in high myopia-induced dark nuclear cataract.

PURPOSE: To assess the etiology of early-onset dark nucleus in high-myopic patients and its relationship with the epigenetic regulation of αA-crystallin (CRYAA). METHODS: We reviewed clinical data from patients who underwent cataract surgery at our center in 2012. Lens epithelial samples were collected during capsulorhexis, whereas young lens epithelium was donated. Cataract type and severity were graded according to the Lens Opacity Classification System III (LOCS III). DNA methylation was analyzed by pyrosequencing the CpG islands of the CRYAA promoter in the following groups: Age-Related Cataract (ARC) Nuclear Color (NC) 2-3; High-Myopic Cataract (HMC) NC2-3; ARC NC5-6; HMC NC5-6; and in young lenses graded NC1. We analyzed CRYAA expression by real-time polymerase chain reaction (PCR), reverse transcription PCR, and immunohistochemistry. RESULTS: The odds ratio of dark nucleus in high-myopic patients was 5.16 (95% confidence interval: 3.98-6.69; p<0.001). CpG islands in lens epithelial CRYAA promoter in the HMC NC5-6 Group exhibited the highest methylation of all the groups, but no statistically significant differences were evident between the HMC NC2-3 and ARC NC2-3 Groups. Likewise, CRYAA mRNA and protein levels in the HMC NC5-6 Group were significantly lower than the ARC NC5-6 Group and high-myopic controls. CONCLUSIONS: High myopia is a risk factor for dark nucleus. Downregulation of CRYAA via the hypermethylation of CpG islands in its promoter could underlie the earlier onset of dark nucleus in high-myopic patients.

catena-Poly[[(2-amino-1,3-benzothia-zole-6-carboxyl-ato-κ(2)O,O')(2,2'-bipyridyl-κ(2)N,N')cadmium]-µ-2-amino-1,3-benzothia-zole-6-carboxyl-ato-κ(3)N(1):O,O'].

In the title coordination polymer, [Cd(C(8)H(5)N(2)O(2)S)(2)(C(10)H(8)N(2))](n), the Cd(II) ion is coordinated by a bidentate 2,2-bipyridyl ligand, two O,O'-chelating 2-amino-1,3-benzothia-zole-6-carboxyl-ate (ABTC) ligands and one N-bonded ABTC ligand. The resulting CdN(3)O(4) coordination polyhedron approximates to a very distorted penta-gonal bipramid with one O and one N atom in axial positions. One of the ABTC ligands is bridging to an adjacent metal atom, generating an infinite chain propagating in [100]. A three-dimensional network is constructed from N-H⋯O and N-H⋯N hydrogen bonds and aromatic π-π stacking inter-actions [centroid-centroid separations = 3.641 (2) and 3.682 (3) Å].

Role of p63/p73 in epithelial remodeling and their response to steroid treatment in nasal polyposis.

BACKGROUND: Nasal polyposis (NP) is recognized as aberrant epithelial remodeling, but the molecular mechanism underlying this process is poorly understood. Two important p53 homologues (p63 and p73) play a key role in orchestrating the epithelial development. OBJECTIVE: We intended to study whether p63 and p73 are involved in the epithelial remodeling seen in patients with NP and their response to oral glucocorticosteroid (GC) treatment. METHODS: Nasal polyp tissues were obtained from 65 patients, and inferior turbinates were obtained from 19 control subjects without NP. Among patients with NP, 20 were treated with oral prednisone, so that 2 sets of polyp biopsy specimens were taken before (GC naive) and after (GC treated) treatment. Immunohistochemistry and quantitative PCR were performed to determine the expression levels of p63 and p73. RESULTS: The increase in p63-positive cell numbers was significant in GC-naive NP epithelium (46%) compared with that seen in control epithelium (5%), and it was positively related to the epithelial hyperplasia in patients with NP. The increase in N-terminal transactivation domain p73-positive cell numbers was found in 27% of GC-naive patients with NP and 16% of control subjects, with no statistical difference. The mRNA expression of both p63 and p73 was significantly upregulated in GC-naive patients with NP versus control subjects, and a positive correlation between the p63 and p73 mRNAs was found in all nasal tissues. Furthermore, the improvement of epithelial structure and reduction of p63 mRNA/protein levels were found in patients with NP after GC treatment. CONCLUSIONS: Our results suggest that the ectopic expression of p63 in multiple cell layers is an important pathologic phenomenon in the epithelial remodeling seen in chronically inflamed airway epithelium (eg, in patients with NP), and its aberrant expression can be suppressed with GC treatment.