Immune modulation of Th1/Th2/Treg/Th17/Th9/Th21 cells in rabbits infected with Eimeria stiedai.

Introduction: Despite long-term integrated control programs for Eimeria stiedai infection in China, hepatic coccidiosis in rabbits persists. Th1, Th2, Th17, Treg, Th9, and Th21 cells are involved in immune responses during pathogen infection. It is unclear whether Th cell subsets are also involved in E. stiedai infection. Their roles in the immunopathology of this infection remain unknown. Therefore, monitoring these T-cell subsets' immune responses during primary infection of E. stiedai at both transcriptional (mRNA) and protein (cytokines) levels is essential. Methods: In experimentally infected New Zealand white rabbits, mRNA expression levels of their transcript-TBX2 (Th1), GATA3 (Th2), RORC (Th17), Foxp3 (Treg), SPI1 (Th9), and BCL6 (Th21)-were evaluated using quantitative real-time polymerase chain reaction (qRT-PCR), whereas Th1 (IFN-g and TNF-a), Th2 (IL4), Th17 (IL17A and IL6), Treg (IL10 and TGF-b1), Th9 (IL9), and Th21 (IL21) cytokines were measured using enzyme-linked immunosorbent assays (ELISAs). Results: We found that levels of TBX2, GATA3, RORC, SPI1, and BCL6 in the livers of infected rabbits were elevated on days 5 and 15 post-infection (PI). The concentrations of their distinctive cytokines IFN-g and TNF-a for Th1, IL4 for Th2, IL17A for Th17, IL9 for Th9, IL21 for Th21, and IL10 for Treg IL10 were also significantly increased on days 5 and 15 PI, respectively (p < 0.05). On day 23 PI, GATA3 with its cytokine IL4, RORC with IL17A, Foxp3 with IL10 and TGF-b1, and SPI1 with IL9 were significantly decreased, but TBX2 with IFN-g and IL6 remained elevated. Discussion: Our findings are the first evidence of Th1/Th2/Treg/Th17/Th9/Th21 changes in E. stiedai-infected rabbits and provide insights into immune regulation mechanisms and possible vaccine development.

Novel lncRNA-prader willi/angelman region RNA, SNRPN neighbour (PWARSN) aggravates tubular epithelial cell pyroptosis by regulating TXNIP via dual way in diabetic kidney disease.

OBJECTIVES: Elevated thioredoxin-interacting protein (TXNIP)-induced pyroptosis contributes to the pathology of diabetic kidney disease (DKD). However, the molecular mechanisms in dysregulated TXNIP in DKD remain largely unclear. MATERIALS AND METHODS: Transcriptomic analysis identified a novel long noncoding RNA-Prader Willi/Angelman region RNA, SNRPN neighbour (PWARSN)-which was highly expressed in a proximal tubular epithelial cell (PTEC) under high glucose conditions. We focused on revealing the functions of PWARSN in regulating TXNIP-mediated pyroptosis in PTECs by targeting PWARSN expression via lentivirus-mediated overexpression and CRISPR-Cas9-based knockout in vitro and overexpressing PWARSN in the renal cortex by AAV-9 targeted injection in vivo. A number of molecular techniques disclosed the mechanisms of PWARSN in regulating TXNIP induced-pyroptosis in DKD. RESULTS: TXNIP-NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome and PTEC pyroptosis were activated in the renal tubules of patients with DKD and in diabetic mice. Then we explored that PWARSN enhanced TXNIP-driven PTECs pyroptosis in vitro and in vivo. Mechanistically, cytoplasmic PWARSN sponged miR-372-3p to promote TXNIP expression. Moreover, nuclear PWARSN interacted and facilitated RNA binding motif protein X-linked (RBMX) degradation through ubiquitination, resulting in the initiation of TXNIP transcription by reducing H3K9me3-enrichment at the TXNIP promoter. Further analysis indicated that PWARSN might be a potential biomarker for DKD. CONCLUSIONS: These findings illustrate distinct dual molecular mechanisms for PWARSN-modulated TXNIP and PTECs pyroptosis in DKD, presenting PWARSN as a promising therapeutic target for DKD.

Two-dimensional shear wave elastography with two different systems for the diagnosis of breast lesions.

BACKGROUND: Two-dimensional (2D) - shear wave elastography (SWE) has made promising advances in the diagnostic of breast lesions. However, few studies have assessed whether the diagnostic effectiveness of different platforms employing 2D-SWE is equal or different. OBJECTIVE: To compare the diagnostic effectiveness of 2D-SWE techniques from two different systems in differentiating malignant breast lesions from benign ones. METHODS: A total of 84 breast lesions were retrospectively analyzed by experienced radiologists using 2D-SWE on two ultrasound systems, i.e. system-1 (LOGIQ E9 system, GE Healthcare, Wauwatosa, WI, USA), and system-2 (Aixplorer US system, SuperSonic Imagine, Aix-en-Provence, France). Qualitative and quantitative parameters including color sign, the maximum elasticity modulus values (E-max), the mean elasticity modulus values (E-mean) and standard deviation (E-sd) of elasticity modulus values in two 2D-SWE systems were analyzed. The diagnostic performance between system-1 and system-2 were evaluated in terms of the areas under the receiver operating characteristic curves (AUROCs). RESULTS: Among the 84 lesions in this study, 66 (78.6%) were benign and 18 (21.4%) were malignant. E-max in system-1 showed the best diagnostic performance with a cut-off value of 174.5 kPa with the associated sensitivity and specificity of 100.0% and 80.3% respectively. Meanwhile, E-sd in system-2 displayed the best diagnostic performance with a cut-off value of 12.7 kPa, with the associated sensitivity and specificity of 94.4% and 80.3% respectively. The diagnostic performance of the two 2D-SWE systems was not statistically different according to receiver operating characteristic curve (ROC) analysis of E-max, E-mean, and E-sd. CONCLUSION: For identifying breast lesions, system-1 and system-2 appear to be similar in diagnostic performance. However, different cut-off values for different parameters might be selected to obtain the best diagnostic performance for the two 2D-SWE systems.

Chemically modified phosphorene as efficient catalyst for hydrogen evolution reaction.

Hydrogen gas produced by electrolysis has been considered as an excellent alternative to fossil fuels. Developing non-noble metal catalysts with high electrocatalytic activities is an effective way to reduce the cost of hydrogen production. Recently, black phosphorus (BP) based materials have been reported to have good potential as electrocatalysts for hydrogen evolution reaction (HER). Herein, we systematically study the catalytic performance of monolayer BP (phosphorene) and several chemically modified phosphorenes (N/S/C/O doping and adsorbed NH2/OH functional groups) for HER on the basis of first-principles calculations. For pristine phosphorene, the armchair edge shows much better catalytic activity than the plane site and zigzag edge. The electronic states of phosphorene near the Fermi level are strongly influenced by chemical modifications. Both of doping heteroatoms into the lattice and introducing NH2/OH functional groups can effectively improve the catalytic performance of the plane site and zigzag edge site, but slightly degrade the armchair edge. These theoretical results shed light on the microscopic understanding of the active sites in BP based electrocatalysts for HER and pave the way for further improving their catalytic performance.

[Glaucoma and Moscow Eye Microsurgery complex (Moroz) keratoprosthesis].

OBJECTIVE: To investigate the relationship between glaucoma and Moscow Eye Microsurgery Complex (Moroz) keratoprosthesis. METHODS: Analysis and retrospective review of consecutive clinical case series. A total of 90 patients had a Moroz keratoprosthesis implantation between Apr, 2000 and Jun, 2011 at our hospital.Fifteen eyes of 15 patients were included in this research. Twelve eyes were identified glaucoma before Moroz keratoprosthesis surgery, and 3 eyes developed glaucoma afterward.Intraocular pressure (IOP), best-corrected visual acuity (BCVA), and treatment of glaucoma were recorded at 1 to 3 months after the Moroz keratoprosthesis implantation. RESULTS: Moroz keratoprosthesis surgery improved vision dramatically in the majority of patients in our study. The most common preoperative corneal diagnosis was alkali burn (8 eyes, 53.3%), which maybe the risk factor of secondary glaucoma after keratoprosthesis surgery.Five of these 15 patients received cyclocryotherapy after the surgery to control elevated IOP. BCVA decreased from 0.8 to 0.2, 0.25 to light perception, 0.3 to 0.1, 0.2 to hand movement, and 0.25 to counting fingers, respectively. CONCLUSIONS: Moroz keratoprosthesis caused glaucoma in only a small number of patients.Visual field, optic nerve appearance and IOP were the main diagnostic modalities after Moroz keratoprosthesis implantation.

Ocular trauma score in transferred fireworks-related ocular injuries: a case series.

BACKGROUND: Our aim was to review the characteristics of transferred fireworks-related ocular damage and to evaluate the prognostic value of the ocular trauma score (OTS) for these injuries. METHODS: This study included 22 transferred patients (19 male, 3 female; mean age 22.6±14.9 years) (25 eyes). The data were retrospectively reviewed, including the characteristics of the geography, types of fireworks, status of injuries, therapeutic procedures, and the best-corrected visual acuity (BCVA). All the injured eyes were classified using the OTS at the time of the initial examination. RESULTS: Twenty eyes (80%) were in OTS category 1, three eyes (12%) were in OTS category 2, and two eyes (8%) were in OTS category 3. All cases received surgical therapy. Six eyes (24%) were enucleated, four (16%) of which achieved an improvement in their final BCVA. There was a statistically significant improvement in final BCVA between OTS category 1 and the other two OTS categories (p=0.016). CONCLUSION: The aforementioned transferred fireworks-related ocular injury cases occurred mainly in young adults, men and active participants, all of which incurred serious vision loss and blindness. The OTS is quite effective for classifying the status and estimating the prognosis of transferred fireworks-related ocular injuries.

Changes in objective vault and effect on vision outcomes after implantable Collamer lens implantation: 1-year follow-up.

PURPOSE: To investigate the changes in vault and the effect on visual outcomes 1 year after implantable Collamer lens (ICL) implantation. METHODS: In this retrospective study, 127 eyes of 66 patients undergoing ICL implantation were examined both before and up to 1 year after the surgery. The examination contents included white-to-white (WTW) diameter, central vault of the ICL (distance between posterior surface of ICL and anterior surface of crystalline lens), refractive error, and wavefront high-order aberration (HOA). All statistical analyses were performed using SPSS 13.0 software. RESULTS: A significant decrease in vault was noted up to 1 month, after which the value stabilized (p=0.001). The moderate vault decreased significantly after the first 3 months postsurgery (paired-samples t test, p<0.05). Low vault showed a tendency to increase and high vault showed a tendency to decrease, but not significantly, over time. There was no statistically significant correlation between the amount of vault and the refractive error (Pearson correlation coefficient R=0.111, p=0.473) and there was a statistically significant correlation between the vault and HOAs (R=0.304, p=0.024). CONCLUSIONS: Implantable Collamer lens vault over the crystalline lens had the tendency toward a slight decrease with time and did not significantly affect the vision outcome 1 year after surgery.

Patients' subjective visual experiences during vitreous surgery under local anesthesia.

OBJECTIVE: To investigate patient's visual experience during vitreous surgery under local anesthesia. METHODS: A prospective and randomized study of 76 patients that underwent vitreous surgeries in our hospital between July 2010 and December 2010 was designed. All patients were interviewed half an hour before and within 1 hour after the surgery using a standardized questionnaire. Basic characteristics of patients and their intraoperative visual experiences like light, color, moving object, pain, fear, and the desire for general anesthesia before and after the surgery were recorded. RESULTS: Sixty patients (78.9%) perceived at least light perception, and 16 patients (21.1%) got no light perception throughout the entire duration of the operation. Forty one patients (53.9%) perceived only light, while 19 patients (25%) experienced moving objects. Thirty nine patients (51.3%) were frightened during their intraoperative visual experiences. Patients with better preoperative best-corrected visual acuity (BCVA) were more likely to be frightened (p=0.002). The frightened experience was affected notably by the different perceptions during operation, moving object (p=0.024), light (p=0.071), and color (p=0.071). Patients below 50 years old, especially from 20 to 30, were more likely to choose general anesthesia after the vitreous surgery. CONCLUSIONS: Most patients (78.9%) experienced at least light perception during the vitreous surgery under local anesthesia. There were no significant differences between the various visual experiences and their basic characteristics. Patients with better preoperative BCVA, perceived moving objects in their visual sensations seemed more likely to be frightened.

[Construction of lentiviral expression vector containing Foxp3 gene and its expression in DC2.4 cell line].

AIM: To construct a Foxp3 lentiviral vector and transfer it into DC2.4 cells, which provides Foxp3+DC cells for further study on its immune modulation. METHODS: We cloned mouse Foxp3 gene into lentiviral vector(pGC-FU) and acquired the plasmid pGC-FU-Foxp3. PCR and sequencing analysis were made for verifying the positive clones. The virus packaging plasmids(pGC-FU-Foxp3, pHelper1.0 and pHelper2.0) were contransfected into 293T cells, and the Lentivirus-Foxp3 was harvested from 293T cells. The Lentivirus-Foxp3 was used to infect DC2.4 cells in vitro and the expression of Foxp3 in infected DC2.4 cells was detected with flow cytometry(FCM). RESULTS: PCR and sequencing revealed that the pGC-FU-Foxp3 plasmid was correctly constructed. The Lentivirus-Foxp3 with a titer of 2×10(8); TU/mL was successfully packaged. Foxp3 expression in DC2.4 cells infected with the Lentivirus-Foxp3 was increased significantly compared with negative control lentivirus. CONCLUSION: The pGC-FU-Foxp3 plasmid has been successfully constructed and the Lentivirus-Foxp3 has been successfully packaged. Foxp3 can be enhanced in DC cells infected with the Lentivirus-Foxp3.

[The observation of the effects of a micromolecular compound J2 on rat corneal allograft using atomic force microscopy].

OBJECTIVE: To study the cell membrane of corneal endothelium with a micromolecular compound J2 in corneal allograft of rat using atomic force microscopy (AFM). METHODS: Cohort study. Subjects were divided into two groups: group A (n = 15): experimental group; group B (n = 15): placebo control group. At the fifth, tenth, fifteen, twentieth, twenty-fifth day after penetrating keratoplasty, the donor implant was separated from receipt bed, one part of which was stained by HE and the others fixed into AFM sample. Amplitude and height images were obtained in the tapping mode with a scan rate of 2 Hz and an integral gain of 0.3 to 0.5. Statistical analysis was performed using single-factor analysis of variance and P value was calculated. RESULTS: The average transplant survival time in group A was (33.12 ± 6.80) d, and those in group B was (18.87 ± 4.19) d. There were significant difference between two group (F = 47.7449, P = 0.00). There were obvious differences on ultrastructure measured by AFM between two groups. At the fifth day after penetrating keratoplasty, regular hexagonal structure of corneal endothelium was observed by AFM in both two group. The diameter of corneal endothelium was about 15 µm, uneven microstructure of cell could be found. The time being, different changes were arose in two group: a clear microstructure could be found in group A, however the microstructure of cell could not be recognized in group B. One way analysis of variance showed that significant differences on parameters (Ra, Rp and Rv) were found between two groups (P < 0.05). At the fifth day after penetrating keratoplasty, group A: Ra (97.64 ± 31.58) nm, Rp (297.79 ± 25.19) nm, Rv (545.55 ± 25.83) nm; group B: Ra (112.61 ± 34.29) nm, Rp (265.06 ± 24.17) nm, Rv (544.41 ± 21.78) nm (Fa = 30.9416, P = 0.0000; Fp = 263.6018, P = 0.0000; Pv = 1.2013, P = 0.2735). At the tenth day after penetrating keratoplasty, group A: Ra (102.98 ± 32.98) nm, Rp (711.38 ± 21.94) nm, Rv (639.89 ± 22.58) nm; group B: Ra (222.85 ± 31.28) nm, Rp (111.22 ± 20.35) nm, Rv (746.49 ± 23.17) nm (Fa = 2086.4535, P = 0.0000; Fp = 53768.4676, P = 0.0000; Pv = 3257.3178, P = 0.0000). At the fifteenth day after penetrating keratoplasty, group A: Ra (87.44 ± 34.97) nm, Rp (344.18 ± 21.09) nm, Rv (482.61 ± 22.27) nm; group B: Ra (197.64 ± 35.72) nm, Rp (510.76 ± 24.98) nm, Rv (545.62 ± 23.17) nm (Fa = 1458.1057, P = 0.0000; Fp = 7788.6963, P = 0.0000; Pv = 1153.2860, P = 0.0000). At the twentieth day after penetrating keratoplasty, group A: Ra (85.85 ± 32.53) nm, Rp (348.69 ± 21.26) nm, Rv (367.65 ± 23.12) nm; group B: Ra (201.36 ± 34.12) nm, Rp (788.58 ± 20.34) nm, Rv (563.33 ± 21.01) nm (Fa = 1801.1215, P = 0.0000; Fp = 67 057.9516, P = 0.0000; Fv = 11 770.2195, P = 0.0000). At the twenty-fifth day after penetrating keratoplasty, group A: Ra (104.97 ± 32.47) nm, Rp (395.05 ± 20.38) nm, Rv (396.17 ± 21.59) nm; group B: Ra (43.85 ± 31.28) nm, Rp (249.88 ± 20.79) nm, Rv (154.88 ± 22.37) nm (Fa = 551.4134, P = 0.0000; Fp = 7458.9255, P = 0.0000; Pv = 18 070.5189, P = 0.0000). CONCLUSIONS: The morphology and ultrastructure of corneal endothelium in group A with J2 were different from group B by observation with AFM. J2 was an effect micromolecular in prevention of corneal allograft rejection.

Experimental study of the mechanism of tolerance induction in dexamethasone-treated dendritic cells.

BACKGROUND: The aim of this study was to investigate the mechanisms underlying tolerance induction of dexamethasone (Dex)-treated dendritic cells (DCs). MATERIAL/METHODS: Well-grown DC2.4 cells were randomly assigned to receive control, 50 µg/L, 100 µg/L, or 200 µg/L of dexamethasone and then were cultured for 6 days. The expressions of CD80, CD86, galectin-9, and PD-L1 on the surface of DC2.4 cells were analyzed with flow cytometry and the level of IL-12 secreted by DC2.4 cells was determined by ELISA. The stimulating activity of DC2.4 cells on allogeneic T cells was assessed with mixed lymphocyte reaction. Dexamethasone-treated DC2.4 cells were co-cultured with allogeneic splenic lymphocytes and the Foxp3 expression in naive T lymphocytes was determined with flow cytometry. RESULTS: Compared with the control group, the expressions of CD80, CD86, galectin-9, and PD-L1 on the surface of DC2.4 cells exposed to different doses of dexamethasone showed no significant changes; however, dexamethasone treatment significantly reduced IL-12 secretion and inhibited DC2.4's stimulation on the proliferation of allogeneic T lymphocytes. Moreover, dexamethasone-treated DC2.4 cells effectively promoted FOXP3 expression in naive T lymphocytes. CONCLUSIONS: DC2.4 is a stable cell line with high expressions of CD80, CD86, and PD-L1. Dexamethasone does not significantly change the cell phenotype of DC2.4 cells, but inhibits the secretion of IL-12 cytokine and attenuates DC2.4's stimulation of the proliferation of allogeneic T cells. Dexamethasone-treated DC2.4 cells also effectively promote FOXP3 expression in naive T lymphocytes.

[The progress of corneal crosslink in corneal disease].

Corneal crosslink through UVA-riboflavin photochemistry has been shown to be an effective treatment for keratoconus, related keratectasias and infectious melting keratitis that did not respond to systemic therapy. It is a new approach to increase the mechanical and biochemical stability of the stromal tissue. The purpose of this paper is to review the basic principle, procedure, laboratory and clinical researches about corneal crosslink. In the current situation, it can state only that UVA-riboflavin induced corneal crosslink leads in the majority of the cases to a halt of the progression, and the complications of the corneal endothelial cells, crystalline lens and retina seem to be rare.

New surgical therapy for conjunctival lymphangiectasia.

AIM: To report 7 cases of conjunctival lymphangiectasia, introduce a simple operation method and discuss the result of the pathological examination. METHODS: Clinical data including medical history, physical examination, previous surgery history were collected in 7 patients with conjunctival lymphangiectasia. Ocular data including visual acuity, Slit-lamp biomicroscopy were released, as were results of histopathology studies. Surgical method was also introduced by video. RESULTS: Seven eyes of 7 patients (4 male and 3 female) with biopsy-proven conjunctival lymphangiectasia underwent the resection. Despite different surgical intervention, the optima method has been founding to treatment conjunctival lymphangiectasia. It is to reduce recurrence that the most concerned problem. In these 7 cases reported, patients kept healthy for 18 months without any one recurrence, whose visual acuity were not affected. Pathological report showed cystic tissue consists of squamous epithelium with significant hyperplasia. CONCLUSION: This operation method is simple and easy to complete, as it is deserved to accept for surgeons and patients on the basis of its advantages.

Estrogen regulates expression of osteoprotegerin and RANKL in human periodontal ligament cells through estrogen receptor beta.

BACKGROUND: The function of periodontal ligament (PDL) cells may be affected by estrogen. PDL cells synthesize the receptor activator of nuclear factor-kappa B ligand (RANKL) and its decoy receptor, namely, osteoprotegerin (OPG), which directly controls osteoclastogenesis. The primary aim of this study was to investigate how estrogen affects the expression of OPG and RANKL in human PDL (hPDL) cells. METHODS: We used a short interfering RNA technique to inhibit estrogen receptor beta (ERbeta) expression in hPDL cells; the cells were cultured with a saturating concentration of 17beta-estradiol (10(7) M) for 48 hours. Changes in the expression of OPG and RANKL were determined by reverse transcription-polymerase chain reaction and supported by Western blot analysis. RESULTS: Estradiol caused an increase in OPG expression and decreased RANKL expression in hPDL cells. However, it had no effect on the expression of OPG and RANKL in hPDL-siERbeta (the short interfering RNA to block ER beta) cells. CONCLUSION: Estrogen may play an important role in exerting antiresorptive effects on alveolar bone, at least in part, by increasing the expression level of OPG versus that of RANKL via ERbeta in hPDL cells.