The OmpR-like Transcription Factor as a Negative Regulator of hrpR/S in Pseudomonas syringae pv. actinidiae.

Bacterial canker of kiwifruit is a devastating disease caused by Pseudomonas syringae pv. actinidiae (Psa). The type III secretion system (T3SS), which translocates effectors into plant cells to subvert plant immunity and promote extracellular bacterial growth, is required for Psa virulence. Despite that the "HrpR/S-HrpL" cascade that sophisticatedly regulates the expression of T3SS and effectors has been well documented, the transcriptional regulators of hrpR/S remain to be determined. In this study, the OmpR-like transcription factor, previously identified by DNA pull-down assay, was found to be involved in the regulation of hrpR/S genes, and its regulatory mechanisms and other functions in Psa were explored through techniques including gene knockout and overexpression, ChIP-seq, and RNA-seq. The OmpR-like transcription factor had binding sites in the promoter region of the hrpR/S, and the transcriptional level of the hrpR/S increased after the deletion of OmpR-like and decreased upon its overexpression in an OmpR-like deletion background. Additionally, OmpR-like overexpression reduced the strain's capacity to form biofilms and lipopolysaccharides, led to its slow growth in King's B medium, and reduced its swimming ability, although there was no significant effect on its pathogenicity against kiwifruit hosts. Our results indicated that OmpR-like directly and negatively regulates the transcription of hrpR/S and may be involved in the regulation of multiple biological processes in Psa. Our results provide a basis for further understanding the transcriptional regulation mechanism of hrpR/S in Psa.

High-Intensity Use of Smartphone Can Significantly Increase the Diagnostic Rate and Severity of Dry Eye.

Purpose: To investigate the effects of high-intensity use of smartphones on ocular surface homeostasis and to explore whether high-intensity use of handheld digital devices can cause false increase of dry eye diagnostic rate. Methods: In this prospective self-control study, 60 subjects (120 eyes) were recruited and asked to read on smartphones provided by the same manufacturer for two consecutive hours. This study was conducted during 8:00 - 10:00 AM to eliminate the influence of digital equipment used the previous day. Ophthalmological examinations [non-invasive tear breakup time (NIBUT), fluorescein breakup time (FBUT), Schirmer I test, corneal fluorescein staining (CFS), bulbar conjunctival redness and meibomian gland (MG) assessment] and a questionnaire survey were conducted before and after the reading test. Based on the collected data, the changes in ocular surface damage and subjective symptoms of the subjects were evaluated, and the differences in the diagnostic rate of dry eye before and after high-intensity use of smartphones were compared. Results: The diagnostic rate of dry eye was sharply increased (61.7% vs. 74.2%). The severity of dry eye also changed significantly, and the moderate and severe degree increased after reading (10% vs. 15%; 5% vs. 10.8%). The aggravated severity subjects had lower MG expressibility and more evident bulbar conjunctival redness compared to the non-aggravated severity subjects. After 2 h of continuous reading, NIBUT-First, NIBUT-Average and FBUT-Average were significantly decreased, while the proportion of BUT ≤ 5 s increased significantly. Non-invasive keratograph tear meniscus height(NIKTMH) decreased significantly compared to the baseline level, while the proportion of NIKTMH<0.20 mm increased significantly. No significant difference was observed in the Schirmer I test and CFS score between the two groups. Compared to the baseline, evident aggravation was observed in bulbar conjunctival redness. The Ocular Surface Disease Index (OSDI) was significantly higher than the baseline after the reading test. Conclusion: Diagnostic indicators related to dry eye are rapidly deteriorating after high-intensity smartphone use, especially those with lower MG expressibility and ocular redness. High-intensity smartphone use can increase the false positive rate of dry eye diagnosis by disturbing ocular surface homeostasis.

Identification of Genetic and Chemical Factors Affecting Type III Secretion System Expression in Pseudomonas syringae pv. actinidiae Biovar 3 Using a Luciferase Reporter Construct.

The type III secretion system (T3SS) is a key factor in the pathogenesis of Pseudomonas syringae pv. actinidiae biovar 3 (Psa3), the causal agent of a global kiwifruit bacterial canker pandemic. To monitor the T3SS expression levels in Psa3, we constructed a luciferase reporter plasmid-expressing HrpAPsa3-NLuc fusion protein. The expression of HrpA-NLuc was induced in hrp-inducing conditions whereas the level of luciferase activity correlated with the expression of hrp/hrc genes in Psa3 confirmed the reliability of the reporter construct. Based on the readout of the NLuc reporter construct, three small molecule compounds 4-methoxy-cinnamic acid, sulforaphane, and ferulic acid were determined as T3SS inhibitors in Psa3, whereas sodium acetate was determined to be a T3SS inducer. Moreover, the aqueous extract of fruit inhibited the accumulation of HrpA-NLuc in Psa3 in medium and in planta. Additionally, the T3SS inhibitors suppress Psa3 virulence, whereas the T3SS inducer promotes Psa3 virulence on kiwifruit. Thus, our findings may provide clues to why the fruit is not infected by Psa3, and the Psa3 T3SS inhibitors have potential as alternatives to current nonspecific antimicrobials for disease management.

In Situ Visualization of Reversible Diels-Alder Reactions with Self-Reporting Aggregation-Induced Emission Luminogens.

The dynamic reversible Diels-Alder (DA) reactions play essential roles in both academic and applied fields. Currently, in situ visualization and direct monitoring of the formation and cleavage of covalent bonds in DA reactions are hampered by finite compatibility and expensive precise instruments, especially limited in solid reactions. We herein report a fluorescence system capable of in situ visualization by naked eyes and monitoring DA/retro-DA reactions. With the fluorescence quenching effect, the synthesized TPEMI could work as an innovative self-indicator for both DA termination and retro-DA occurrence. The fluorescence increases during DA reactions, and the mechanism is investigated to establish qualitative and quantitative relations. Besides rapid screening of reaction conditions and monitoring of DA exchange processes, the TPEMI fluorescence system can visualize heterogeneous and solid-state reactions with the AIE character. The TPEMI platform is expected to offer novel insights into reversible DA processes and dynamic covalent chemistry.

Catechol Moiety Integrated Tri-Aryl Type AIEgen for Visual and Quantitative Boronic Acid Detection.

Novel functional AIEgen based on three compact bound aryl skeletons is designed and synthesized. This tri-aryl type luminogen (TA-Catechol) embedded with catechol moiety responds rapidly to series of boronic acids. Real-time visual and quantitative dual-mode detection method is established for the first time with modest precision and low detection limit (8.0 µM). Detailed mechanistic discussion identifies tetra-coordinated boronic species as the key intermediate within sensing procedure. Wide range of organic boronic acids compatible with this strategy is displayed which is promising in high throughput screening technology. Furthermore, solid-state sensing capability of TA-Catechol is also demonstrated.

Prospective trial of a 2940 nm Er:YAG laser for the treatment of meibomian gland dysfunction.

PURPOSE: The primary objective was to evaluate the efficacy and safety of Er:YAG laser treatment for meibomian gland dysfunction (MGD) in a prospective study. METHODS: A total of 128 eyes from 64 patients with MGD were enrolled to receive either three Er:YAG laser treatments with meibomian gland expression (MGX) or MGX-alone treatment sessions at 3-week intervals. The Standard Patient Evaluation of Eye Dryness (SPEED) validated questionnaire; fluorescein breakup time of the tear film (FBUT); corneal fluorescein staining (CFS); lid margin abnormalities; meibomian gland morphology (meiboscore); lower tear meniscus height (TMH); and assessment of 15 meibomian glands in the lower eyelids, including total meibomian gland secretion quality (TMGS), the number of glands secreting any liquid (GSAL), and the number of glands yielding optimal clear liquid secretion (GYCL), were assessed at day (D)0, D21, D42, and D63 for the Er:YAG-MGX group and D0 and D63 for the MGX group. RESULTS: At D63, significant decreases in SPEED scores and lid margin abnormalities as well as significant increases in FBUT, TMGS, and GSAL were observed in both groups (all p < 0.05). The Er:YAG-MGX group showed a significantly better improvement in SPEED scores, TMGS, and GYCL than the MGX group (all p < 0.05). CONCLUSION: Although preliminary, the study results of Er:YAG laser treatment for dry eye syndrome caused by MGD are promising. Er:YAG laser treatment may be a new direction for managing MGD. TRIAL REGISTRATION: The study was registered at www.chictr.org.cn : ChiCTR1900026004.

Intrinsic hydroquinone-functionalized aggregation-induced emission core shows redox and pH sensitivity.

Aggregation-induced emission (AIE) fluorophores exhibit strong fluorescence in an aggregated state but emit no or weak fluorescence in dilute solutions. This emerging class of AIE optical materials comprise a variety of functionalities. Here an AIE luminescence core, 1-hydroquinol-1,2,2-triphenylethene (HQTPE), has been designed and synthesized. This AIE core is simple but is fundamentally important to chemistry because of its intrinsic redox and pH activities. The incorporation of hydroquinone (HQ) moiety into a common AIE core tetraphenylethene (TPE) yields HQTPE with unique fluorescent properties like nonlinear self-quenching over most other AIE-active fluorophores (AIEgens) so far reported. There are differences of photochemical properties between HQTPE, 1-benzoquinol-1,2,2-triphenylethene (QTPE, the oxidized counterpart) and its anions. Interestingly, as the solution concentration is increased, AIEgen HQTPE shows stronger fluorescence but QTPE exhibits rapid quenching of fluorescence in a nonlinear fashion, which are in agreement with theoretical studies. The fluorescence of HQTPE is also highly dependent on the pH value of media. We have further explored HQTPE as an ultrasensitive redox probe and efficient deoxidizer, which could lead to potential applications in health care, food security, environmental monitoring, optic and electronic devices.

Neutrophil extracellular traps may have a dual role in Pseudomonas aeruginosa keratitis.

Pseudomonas aeruginosa (P. aeruginosa) keratitis is a sight-threatening and rapidly progressive corneal disease. Neutrophils and neutrophil extracellular traps (NETs) are widely thought to play a vital role in hosts' immune defenses against bacteria, such as P. aeruginosa. The present study aimed to investigate the dynamics of the formation and the role of NETs in P. aeruginosa keratitis. First, scratched corneas of mice models were treated with 1 × 108 colony-forming units (CFU)/ml of P. aeruginosa suspension or normal saline (NS). Second, after 48 h postinfection, the infected corneas were treated with TobraDex, Tobrex, 0.1% dexamethasone, or NS four times a day, respectively. Clinical examination, hematoxylin and eosin (H&E) staining, immunofluorescence staining, scanning electron microscopy, and bacterial burden testing were performed on the corneas. Tobrex reduced neutrophil infiltration and corneal P. aeruginosa burden. Dexamethasone reduced NETs, bacterial burden, and severe neutrophil infiltration. TobraDex produced a greater reduction in the amount of neutrophils, NETs, and bacterial burden and the results of Tobrex-treated group were between them. These findings corresponded with the clinical findings that TobraDex- and Tobrex-treated mice exhibited slight corneal damage, while dexamethasone-treated mice exhibited very severe corneal damage. Cumulatively, our data suggest that NETs may play a dual role of infection control and corneal damage in P. aeruginosa keratitis. Furthermore, combination treatment targeting NET formation and bacteria may serve as a way of improving the clinical outcomes of bacterial keratitis.

Influence of Pseudomonas autoinducer N-3-oxododecanoyl homoserine lactone on human corneal epithelial cells.

The quorum-sensing (QS) signaling-dependent extracellular virulence factors of Pseudomonas aeruginosa can cause infections such as P. aeruginosa keratitis. P. aeruginosa communicates by secreting and sensing small chemical molecules called autoinducers in QS system. The key QS signal molecule, N-3-oxododecanoyl-homoserine lactone (3OC12HSL), can affect the behavior of host cells and initiate immune response. In this report we investigated the influence of 3OC12HSL on human corneal epithelial cells (HCECs) and the mechanisms of 3OC12HSL on activated toll-like receptor 2 (TLR2)-dependent interleukin-8 (IL-8) secretion in HCECs. Cells were cultured under different concentrations of 3OC12HSL. Cell viability was assessed using Crystal violet staining and the cell counting kit-8 assay. We demonstrated the administration of 3OC12HSL decreased HCEC viability and survival in a concentration- and time-dependent manner. At high concentrations, 3OC12HSL rapidly promoted a time-dependent increase in the expressions of TLR2 and TLR4. It was found that the nuclear translocation and expression of nuclear factor-κB (NF-κB) were also increased in response to 3OC12HSL treatment. The significantly elevated expressions of TLR2, TLR4, and NF-κB, encouraged us to further test their mechanisms that cause inflammatory response. Among the inflammatory factors examined (IL-6, IL-8, IL-10, and TNF-α), we found that IL-8 was significantly increased after treatment with 3OC12HSL and its expression was inhibited when TLR2 was specifically blocked or silenced. These results indicated that the QS signaling molecule 3OC12HSL could be recognized by the host innate immune system in HCECs. This recognition then triggered an immune inflammatory response involving the activation of TLR2 and an increase in expression of IL-8. This crosstalk between 3OC12HSL and host immunity in HCECs contributes to the development and progression of P. aeruginosa keratitis.

Neutrophil extracellular traps promote corneal neovascularization-induced by alkali burn.

OBJECTIVES: To investigate the effects of neutrophil extracellular traps (NETs) on angiogenesis in vitro and in vivo and the regulatory role of mammalian target of rapamycin (mTOR) activity in it. METHODS: The regulatory role of mTOR in NETs formation was explored. In vitro, human neutrophils were pretreated with rapamycin. NETs formation was measured using immunofluorescence staining of NETs markers, SYTOX Green and PicoGreen after NaOH stimulation. In vivo, mice were treated with rapamycin, and NETs formation in cornea was measured using immunofluorescence staining 7 days after alkali burn. Then, the effects of NETs on angiogenesis were investigated. In vitro, human neutrophils were treated with DNase I or rapamycin. NETs were isolated after NaOH stimulation and the isolated NETs were co-culture with human umbilical vein endothelial cells (HUVECs). HUVECs migration, proliferation, and inflammatory activation were measured. In vivo, mice were injected subconjunctivally with supernatant containing NETs. Corneal neovascularization was visualized by immunofluorescence staining. RESULTS: NETs structures can be observed in NaOH-stimulated neutrophils and alkali-burned mouse cornea compared with normal group. Treated with rapamycin enhanced NETs formation in response to NaOH management compared with DMSO control in vitro and in vivo. NETs increased the migration, proliferation and inflammatory activation of HUVECs, and subconjunctival injection of NETs promoted inflammatory and angiogenic response in corneal alkali burn model. CONCLUSIONS: NETs formation can be triggered by NaOH stimulation. mTOR activity has a negative regulatory effect on NETs formation. NETs promoted angiogenic responses and inflammatory activation of HUVECs and increased corneal neovascularization and inflammatory response.

Preparation, Characterization and Diagnostic Valuation of Two Novel Anti-HPV16 E7 Oncoprotein Monoclonal Antibodies.

At present, the clinical detection method of human papillomavirus (HPV) is mainly based on the PCR method. However, this method can only be used to detect HPV DNA and HPV types, and cannot be used to accurately predict cervical cancer. HPV16 E7 is an oncoprotein selectively expressed in cervical cancers. In this study, we prepared an HPV16 E7-histidine (HIS) fusion oncoprotein by using a prokaryotic expression and gained several mouse anti-HPV16 E7-HIS fusion oncoprotein monoclonal antibodies (mAbs) by using hybridoma technology. Two mAbs, 69E2 (IgG2a) and 79A11 (IgM), were identified. Immunocytochemistry, immunofluorescence, immunohistochemistry, and Western blot were used to characterize the specificity of these mAbs. The sequences of the nucleotide bases and predicted amino acids of the 69E2 and 79A11 antibodies showed that they were novel antibodies. Indirect enzyme-linked immunosorbent assay (ELISA) with overlapping peptides, indirect competitive ELISA, and 3D structural modeling showed that mAbs 69E2 and 79A11 specifically bound to the three exposed peptides of the HPV16 E7 (HPV16 E749-66, HPV16 E773-85, and HPV16 E791-97). We used these two antibodies (79A11 as a capture antibody and 69E2 as a detection antibody) to establish a double-antibody sandwich ELISA based on a horseradish peroxidase (HRP)-labeled mAb and tetramethylbenzidine (TMB) detection system for quantitative detection of the HPV16 E7-HIS fusion oncoprotein, however, it was not ideal. Then we established a chemiluminescence immunoassay based on a labeled streptavidin-biotin (LSAB)-ELISA method and luminol detection system-this was sufficient for quantitative detection of the HPV16 E7-HIS fusion oncogenic protein in ng levels and was suitable for the detection of HPV16-positive cervical carcinoma tissues. Collectively, we obtained two novel mouse anti-HPV16 E7 oncoprotein mAbs and established an LSAB-lumino-dual-antibody sandwich ELISA method for the detection of the HPV16 E7-HIS fusion oncogenic protein, which might be a promising method for the diagnosis of HPV16-type cervical cancers in the early stage.

Immunodiagnosis and Immunotherapeutics Based on Human Papillomavirus for HPV-Induced Cancers.

Infection with human papillomavirus (HPV) is one of the main causes of malignant neoplasms, especially cervical, anogenital, and oropharyngeal cancers. Although we have developed preventive vaccines that can protect from HPV infection, there are still many new cases of HPV-related cancers worldwide. Early diagnosis and therapy are therefore important for the treatment of these diseases. As HPVs are the major contributors to these cancers, it is reasonable to develop reagents, kits, or devices to detect and eliminate HPVs for early diagnosis and therapeutics. Immunological methods are precise strategies that are promising for the accurate detection and blockade of HPVs. During the last decades, the mechanism of how HPVs induce neoplasms has been extensively elucidated, and several oncogenic HPV early proteins, including E5, E6, and E7, have been shown to be positively related to the oncogenesis and malignancy of HPV-induced cancers. These oncoproteins are promising biomarkers for diagnosis and as targets for the therapeutics of HPV-related cancers. Importantly, many specific monoclonal antibodies (mAbs), or newly designed antibody mimics, as well as new immunological kits, devices, and reagents have been developed for both the immunodiagnosis and immunotherapeutics of HPV-induced cancers. In the current review, we summarize the research progress in the immunodiagnosis and immunotherapeutics based on HPV for HPV-induced cancers. In particular, we depict the most promising serological methods for the detection of HPV infection and several therapeutical immunotherapeutics based on HPV, using immunological tools, including native mAbs, radio-labelled mAbs, affitoxins (affibody-linked toxins), intracellular single-chain antibodies (scFvs), nanobodies, therapeutical vaccines, and T-cell-based therapies. Our review aims to provide new clues for researchers to develop novel strategies and methods for the diagnosis and treatment of HPV-induced tumors.

Glucocorticoids May Exacerbate Fungal Keratitis by Increasing Fungal Aggressivity and Inhibiting the Formation of Neutrophil Extracellular Traps.

Purpose: To evaluate whether glucocorticoids affect the prognosis of fungal keratitis by inhibiting the formation of neutrophil extracellular traps (NETs).Methods: A mouse model of Candida albicans (C.albicans) keratitis was established. Animals were randomly assigned to treatment with 0.1% dexamethasone (DXM) eye drops and normal saline (3 times each day for 3 days). The effects of DXM on fungal keratitis were assessed using clinical scores, immunofluorescence staining, histopathological examination, scanning electron microscopy (SEM), and pathogen burden assay. All the analyses were performed using SPSS software version 17.0 (Chicago, IL).Results: NETs formation was noteworthy in the cornea lesions of fungal keratitis. The clinical score of the DXM-treated group was significantly higher than that of the control group (P < .05). During the measured period, corneas from DXM-treated group contained more C.albicans than those from the control group by histology and pathogen burden assay. Compared with the control group, the DXM treatment group had a higher depth of infiltration of C.albicans. Histological and immunofluorescence staining showed that there were fewer neutrophils in the cornea focus of DXM-treated group (P < .05）, and the number of NETs formed in scrapings from control group was higher than that in the DXM treatment group on day 3 (P < .05, Z = -3.56)) and day 5 (P < .05, Z = -3.69). In a similar amount of cell scraping, the NETs of neutrophils formation from the DXM-treated group were also less than that from the control group.Conclusion: Our results indicated that NETs were involved in the immune response in C.albicans keratitis. Glucocorticoids may exacerbate fungal keratitis not only by increasing fungal aggressivity and reducing the infiltration of neutrophils but also by inhibiting the formation of NETs.

Demethylzeylasteral inhibits glioma growth by regulating the miR-30e-5p/MYBL2 axis.

Glioma is the most common and malignant form of primary brain tumour, and is characterised by high proliferation and extensive invasion and neurological destruction. Demethylzeylasteral (T-96), which is extracted from Tripterygium wilfordii, is considered to have immunosuppressive, anti-inflammatory and anti-angiogenic effects. Here, the anti-tumour effect of T-96 on glioma was evaluated. Our results demonstrated that T-96 significantly inhibited glioma cell growth and induced cell cycle arrest in G1 phase but did not induce apoptosis. Cell invasion and migration were dramatically suppressed after treatment with T-96. Almost all genes related to cell cycle and DNA replication were downregulated after treatment with T-96. Our results showed that miR-30e-5p was noticeably upregulated after T-96 treatment, and MYBL2, which is involved in cell cycle progression and is a target gene of miR-30e-5p, was significantly reduced in synchrony. Overexpression of MYBL2 partially rescued the T-96-induced inhibition of cell growth and proliferation. Moreover, a miR-30e-5p antagomir significantly reduced the upregulation of miR-30e-5p expression induced by T-96, leading to recovery of MYBL2 expression, and partially rescued the T-96-induced inhibition of cell growth and proliferation. More important, T-96 effectively upregulated miR-30e-5p expression and downregulated MYBL2 expression, thus inhibiting LN-229 cell tumour growth in a mouse model. These results indicated that T-96 might inhibit glioma cell growth by regulating the miR-30e-5p/MYBL2 axis. Our study demonstrated that T-96 might act as a promising agent for malignant glioma therapy.

Evaluation of artificial tears on cornea epithelium healing.

AIM: To observe the efficacy of different artificial eye drops on corneal epithelium healing in rabbit. METHODS: Thirty-five rabbits with 6 mm diameter central corneal epithelium removed were randomly assigned to six groups: 0.9% normal saline (NS) group, 0.1% hyaluronate (HA) group, 0.3% HA group, Tears Naturale Free® (TNF) group, 0.4% polyethylene glycol (PEG) group, 0.5% carboxymethyl cellulose (CMC) group and blank control group. Treatments were administered topically four times daily. Corneal epithelium healing was evaluated by the percentage reduction in wound area at 24, 36, 48, 60, and 72h after removal of the corneal epithelium. Cornea re-epithelialization was also assessed by histological analysis and electron microscopy. RESULTS: All corneal wounds completely re-epithelialized in less than 72h. The average re-epithelialization time was 47.61±4.25h in the 0.3% HA group and 49.72±1.05h in the 0.9% NS group, followed by 0.1% HA, TNF, 0.4% PEG, 0.5% CMC, and lastly by the control group. Compared to the control group, there were significant differences among 0.3% HA, 0.9% NS, PEG, and TNF (P<0.05) groups. At the first 24h, re-epithelialization at the 0.3% HA, TNF, and 0.9% NS treatment groups were significantly faster than the other groups. At 48h post-wounding, corneal epithelium is nearly completing re-epithelialization at 0.3% HA and 0.9% NS treatment groups. Electron microscopy revealed that there were a large number of vacuoles in the cells of the 0.9% NS group at 72h. CONCLUSION: Artificial tears promote corneal re-epithelium varied in the efficacy. Obviously, all artificial eye drops better than blank group. In the process of corneal healing, corneal epithelium cells suffered from hypoxia caused by NS.

Cobalt-Catalyzed Cross-Dehydrogenative Coupling Reaction between Unactivated C(sp2)-H and C(sp3)-H Bonds.

Catalytic oxidative cross-dehydrogenative coupling between unactivated C(sp2)-H and C(sp3)-H bonds is achieved by the cobalt-catalyzed o-alkylation reaction of aromatic carboxamides containing (pyridin-2-yl)isopropyl amine (PIP-NH2) as a N,N-bidentate directing group. Many different C(sp3)-H bonds in alkanes, toluene derivatives and even in the α-position of ethers and thioethers can be used as coupling partners. This method has a broad substrate scope and the tolerance of various functional groups.

Serum Vitamin A Levels May Affect the Severity of Ocular Graft-versus-Host Disease.

Allogeneic hematopoietic stem cell transplantation (HSCT) is a well-established therapeutic option for a range of inherited and acquired hematological disorders. However, graft-versus-host disease (GVHD) remains the leading cause of non-relapse mortality in allogeneic HSCT recipients. Ocular involvement occurs in up to 80% of chronic GVHD patients. In our cases, the diagnosis of vitamin A deficiency was suspected for GVHD patients. Serum vitamin A measurements were conducted to confirm clinical suspicions. Our study revealed significant decrease in serum levels of vitamin A in chronic liver GVHD patients. Although there have been many studies evaluating ocular manifestations in patients with GVHD, the present study is, to our knowledge, the first to study the relationship between vitamin A and ocular manifestations of GVHD in humans. Our data suggest that vitamin A deficiency affects the severity of ocular GVHD in adults.

Dexamethasone Inhibits S. aureus-Induced Neutrophil Extracellular Pathogen-Killing Mechanism, Possibly through Toll-Like Receptor Regulation.

Neutrophils release neutrophil extracellular traps (NETs) in a pathogen-killing process called NETosis. Excessive NETs formation, however, is implicated in disease pathogenesis. Therefore, to understand how NETosis is regulated, we examined the effect of dexamethasone (DXM), an anti-inflammatory drug, on this process and the role of toll-like receptors (TLRs). We stimulated human neutrophils with phorbol 12-myristate 13-acetate (PMA) or Staphylococcus aureus (S. aureus) and quantified NETs formation. We also examined the effect of DXM on the bactericidal effect of NETs and the role of reactive oxygen species (ROS) and nuclear factor (NF)-κB in DXM-regulated NETosis. DXM significantly inhibited S. aureus-induced NETosis and extracellular bacterial killing. ROS production and NF-κB activation were not involved in DXM-regulated NETosis. TLR2 and TLR4, but not TLR5 or TLR6, modified S. aureus-induced NETs formation. Neither DXM nor TLRs were involved in PMA-induced NETosis. Furthermore, TLR2 and TLR4 agonists rescued DXM-inhibited NETosis, and neither TLR2 nor TLR4 antagonists could further inhibit NETosis reduction induced by DXM, indicating that DXM may inhibit NETosis by regulating TLR2 and TLR4. In conclusion, the mechanisms of S. aureus- and PMA-induced NETosis are different. DXM decreases NETs formation independently of oxidant production and NF-κB phosphorylation and possibly via a TLR-dependent mechanism.

A treatment method for chronic suppurative lacrimal canaliculitis using chalazion forceps.

PURPOSE: The purpose of this study is to evaluate the effectiveness of chronic suppurative lacrimal canaliculitis treatment using chalazion forceps. PATIENTS AND METHODS: A prospective study was performed on consecutive patients who accepted the aid of chalazion forceps to treat chronic suppurative lacrimal canaliculitis. Two different treatment methods using chalazion forceps were performed according to the degree of lacrimal canaliculitis. Postoperatively, the patients received 0.5% levofloxacin eye drops four times per day and 0.5 g oral levofloxacin tablets once per day for 4 days. The follow-up period was more than 3 months. Lacrimal irrigation, the condition of the lacrimal punctum, and patients' symptoms were carefully evaluated. RESULTS: In total, 32 patients met the criteria for chronic suppurative lacrimal canaliculitis. Included were 6 males and 26 females. Their average age was 51.7 ± 14.9 years (range; 19-80 years), and all had unilateral canaliculitis. The mean duration of the symptoms was 18.9 ± 9.8 months (range; 3-48 months). The mean follow-up time was 14.7 ± 7.8 months. The signs and symptoms resolved completely in all patients within 15 days, and no recurrence was observed. No patients reported epiphora after the treatment. CONCLUSIONS: The use of chalazion forceps is effective in treating chronic suppurative lacrimal canaliculitis. The forceps may offer an alternative treatment technology in the management of suppurative lacrimal canaliculitis.

Cobalt-Catalyzed C(sp(2) )-H Methylation by using Dicumyl Peroxide as both the Methylating Reagent and Hydrogen Acceptor.

The first cobalt-catalyzed direct methylation of a C(sp(2) )-H bond using dicumyl peroxide (DCP) as both the methylating reagent and hydrogen acceptor has been established. The reaction proceeded without the use of any additives, and was proven to be applicable to various amides bearing a 2-pyridinylisopropyl (PIP) directing group, providing an efficient access to o-methyl aryl amides with high functional-group tolerance. Preliminary mechanistic studies suggest a radical process would be involved in the catalytic process.