Screening of potential drugs for the treatment of diabetic kidney disease using single-cell transcriptome sequencing and connectivity map data.

OBJECTIVE: To explore the feasibility of screening potential drugs for the treatment of diabetic kidney disease (DKD) using a single-cell transcriptome sequencing dataset and Connectivity Map (CMap) database screening. METHODS: A DKD single-nucleus transcriptome sequencing dataset was analyzed using Seurat 4.0 to obtain specific podocyte subclusters and differentially expressed genes (DEGs) related to DKD. These DEGs were subsequently subjected to a search against the CMap database to screen for drug candidates. Cell and animal experiments were conducted to evaluate the efficacy of the top 3 drug candidates. RESULTS: Initially, we analyzed the DKD single-nucleus transcriptome sequencing dataset to obtain intrinsic renal cells such as podocytes, endothelial cells, mesangial cells, proximal tubular cells, collecting duct cells and immune cells. Podocytes were further divided into four subclusters, among which the proportion of POD_1 podcytes was significantly greater in DKD kidneys than in control kidneys (34.0 % vs. 3.4 %). The CMap database was searched using the identified DEGs in the POD_1 subcluster, and the drugs, including tozasertib, paroxetine, and xylazine, were obtained. Cell-based experiments showed that tozasertib, paroxetine and xylazine had no significant podocyte toxicity in the concentration range of 0.01-50 µM. Tozasertib, paroxetine, and xylazine all reversed the advanced glycation end products (AGEs)-induced decrease in podocyte marker levels, but the effect of paroxetine was more prominent. Animal experiments showed that paroxetine decreased urine ALB/Cr levels in DKD model mice by approximately 51.5 % (115.7 mg/g vs. 238.8 mg/g, P < 0.05). Histopathological assessment revealed that paroxetine attenuated basement membrane thickening, restored the number of foot processes of podocytes, and reduced foot process fusion. In addition, paroxetine also attenuated renal tubular-interstitial fibrosis. Mechanistically, paroxetine inhibited the expression of GRK2 and NLRP3, decreased the phosphorylation level of p65, restored NRF2 expression, and relieved inflammation and oxidative stress. CONCLUSION: This strategy based on single-cell transcriptome sequencing and CMap data can facilitate the identification and aid the rapid development of clinical DKD drugs. Paroxetine, screened by this strategy, has excellent renoprotective effects.

Meta-Analysis of the Association between Fibroblast Growth Factor 23 and Mortality and Cardiovascular Events in Hemodialysis Patients.

INTRODUCTION: This study was conducted to ensure the correlation between fibroblast growth factor 23 (FGF23) and death and cardiovascular events in hemodialysis patients. METHODS: We conducted a literature search of PubMed database to April 2018 for relevant articles that reported the association between FGF23 and risk of all-cause mortality and cardiovascular disease (CVD) events. The meta-analysis was performed using the Revman 5.3 software. RESULTS: A total of 7 articles were included, and all reported mortality in hemodialysis patients, and 3 reported cardiovascular events in hemodialysis patients. Since the current reagent can detect C-terminal FGF23 (cFGF23) and intact-FGF23 (iFGF23), we discuss the association between cFGF23 and iFGF23 and death and cardiovascular events in hemodialysis patients. The correlation between serum iFGF23 levels and death in hemodialysis patients: high levels of iFGF23 vs. low levels of iFGF23: RR 1.14, 95% CI (1.01-1.30), p = 0.04 (I2 = 0%, p = 0.38). The correlation between serum C-terminal levels and death in hemodialysis patients: high levels of cFGF23 versus low levels of cFGF23: RR 1.39, 95% CI (1.21-1.59), p < 0.001 (I2 = 2%, p = 0.38). The correlation of serum iFGF23 levels with cardiovascular events: high levels of iFGF23 versus low levels of iFGF23: RR 1.21, 95% CI (1.13-1.30), p < 0.001 (I2 = 0%, p = 0.49). A paper has reported the association between cFGF23 and CVD events, so we did not conduct meta-analysis. CONCLUSIONS: Elevated serum FGF23 levels are positively associated with all-cause mortality and cardiovascular events in hemodialysis patients, with a 14 or 39% increase in all-cause mortality and a 21% increased risk of cardiovascular events.

Metastatic orbital tumors in southern China during an 18-year period.

OBJECTIVE: To determine the primary sites, clinical features, treatment, and outcome of 46 patients with cancer metastatic to the orbits in southern China. METHODS: Retrospective review of medical records at a tertiary center in southern China between Jan 1, 1991 and Dec 31, 2008. RESULTS: The study included 35 males and 11 females, with an age range of 3-78 years (average 47 years). All of the patients came from southern China. The left orbit was affected more frequently than the right orbit (3:2), and no cases were bilaterally involved. The most common primary cancer that metastasizes to the orbit is nasopharyngeal carcinoma (30.34%), followed by lung cancer (8.70%), liver cancer (6.52%), breast carcinoma (4.35%), renal cell cancer (4.35%), medullary thyroid carcinoma (2.17%), lacrimal gland carcinoma (2.17%), stomach carcinoma (2.17%), mediastinum carcinoma (2.17%), other forms (6.52%), and unknown carcinomas (30.34%). Orbital metastasis was the first sign of systemic cancer in 24 patients (52.17%). The five most common clinical findings at the initial examination were abnormal eye position (65.22%), proptosis (63.04%), increased orbital pressure (54.35%), diplopia (54.35%), and limited ocular motility (52.17%), followed by swollen eyelid (50%), visible mass or swelling (46%), blurred vision (41%), conjunctival hyperemia, and edema (35%), pain (28%), blepharoptosis (20%), papilledema (17%), and vasodilatation of the retina (7%). Primary treatment included surgical excision, radiotherapy, systemic chemotherapy, and observation. Among the patients (78% of 46) with sufficient follow-up time (mean 20.7 months), 30.56% of them died of disseminated metastasis. Twenty-five percent of them were in remission with no active metastases and 27.78% of them were living with active metastases. Overall, the mean survival time was 18 months (range, 5-38 months) after diagnosis of orbital metastasis. CONCLUSIONS: The most common primary cancer that metastasizes to the orbit is nasopharyngeal carcinoma (30.34%), followed by lung cancer (8.70%) and liver cancer (6.52%) in southern China. Orbital metastasis can display a variety of clinical features and should be considered in patients with known systemic cancer. The systemic prognosis is generally poor.

Early changes in ocular surface and tear inflammatory mediators after small-incision lenticule extraction and femtosecond laser-assisted laser in situ keratomileusis.

PURPOSE: To characterize the early ocular-surface changes or tear inflammatory-mediators levels following small-incision lenticule extraction (ReLEx smile) and femtosecond laser-assisted laser in situ keratomileusis (FS-LASIK). METHODS: Forty-seven myopic subjects were recruited for this prospective study. Fifteen underwent ReLEx smile and thirty-two underwent FS-LASIK. Corneal fluorescein (FL) staining, tear break-up time (TBUT), Schirmer I test (SIT), ocular surface disease index (OSDI) and central corneal sensitivity were evaluated in all participants. Tears were collected and analyzed for interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), nerve growth factor (NGF) and intercellular adhesion molecule-1 (ICAM-1) levels using multiplex magnetic beads. All measurements were preformed preoperatively and 1 day, 1 week, 1 month and 3 months postoperatively. RESULTS: FL scores in ReLEx smile group were lower than those of FS-LASIK group 1 week postoperatively (P = 0.010). Compared to the FS-LASIK group, longer TBUT were observed in ReLEx smile group 1 month (P = 0.029) and 3 months (P = 0.045) postoperatively. No significant differences were found in tear secretion for the two groups (P>0.05). OSDI scores were higher in FS-LASIK group 1 month after surgery (P = 0.020). Higher central corneal sensitivity was observed in ReLEx smile group 1 week, 1 month and 3 months (P<0.05) postoperatively. Compared to FS-LASIK group, lower and faster recovery of IL-6 and NGF levels in tears was observed in ReLEx smile group postoperatively (P<0.05). Tears TNF-α and ICAM-1 concentrations were not significantly different between the two groups at any follow-up time (P>0.05). Moreover, IL-6 and NGF levels correlated with ocular surface changes after ReLEx smile or FS-LASIK. CONCLUSIONS: In the early postoperative period, ReLEx smile results in milder ocular surface changes than FS-LASIK. Furthermore, the tear inflammatory mediators IL-6 and NGF may play a crucial role in the ocular surface healing process following ReLEx smile and FS-LASIK.

Effects of silicone hydrogel contact lenses on ocular surface after Sub-Bowman's Keratomileusis.

PURPOSE: To evaluate the efficacy of silicone hydrogel contact lenses on ocular surface after Sub-Bowman Keratomileusis (SBK). MATERIALS AND METHODS: Forty-six patients suffered from myopia underwent a bilateral SBK. Post-operatively, one eye of each patient wore a PureVision contact lens for 24 h as a treated eye and the contralateral eye was as a blank control. Afterwards, corneal fluorescein (FL) staining, tear break-up time (TBUT), schirmer I test (SIT), central corneal thickness (CCT), ocular surface disease index (OSDI), corneal hysteresis (CH), corneal resistance factor (CRF) and corneal flap complications were assessed 1 d (except for CH and CRF), 1 week, 1 month and 3 months postoperatively. RESULTS: Following SBK, in contrast to the control, corneal fluorescein staining of treated eyes were significantly reduced and tear break-up time of treated eyes were significantly improved at 1 d and 1 week after SBK. However, Schirmer I test of treated and control eyes were not different after SBK. Central corneal thickness of treated eyes were significantly thinner than that of control at 1 d after SBK, however, there were no differences at other time points. Ocular surface disease index of treated eyes were obviously alleviated more than that of control at 1 d after SBK, but no differences were found at other visits. Moreover, Corneal hysteresis and corneal resistance factor of treated and un-treated eyes were not different after surgery. And also the rate of corneal flap complications were not different between treated and control eyes after SBK. CONCLUSIONS: Silicone hydrogel contact lenses played a positive role in accelerating corneal epithelial healing, enhancing tear film stability and reducing discomfort of patients in the early stage after SBK.

Changes in corneal biometry and the associated histology in rhesus monkeys wearing orthokeratology contact lenses.

PURPOSE: This study compared the effect of orthokeratology (OK) lens wear on corneal topography and microstructure between the 1-night and 30-night wearers to predict any adverse responses of the eyes to long-term OK lens wear. METHODS: Nine rhesus monkeys with one eye wearing an OK lens and the fellow eye wearing a rigid gas permeable (RGP) lens were used in this study. The lenses were worn for 11 hours, with the measurement of corneal curvature and thickness and axial components of the eye at 0 night (n = 9), 1 night (n = 9), and 30 nights (n = 6). Histology was performed at 1 and 30 nights of lens wear. RESULTS: Corneal surface power in the 3-mm central region was reduced in the OK eye when compared with the RGP eye (P < 0.05). Central corneal thickness was only reduced in 30 nights of OK lens wear (P < 0.05). Under light microscopy, the 1-night OK or RGP eyes showed normal morphology in all layers of the cornea. The 30-night OK eyes showed a thinned central corneal epithelium and a thickened but less stratified paracentral corneal epithelium when compared with the 30-night RGP eyes. Under electron microscopy, intercellular junctions of corneal epithelium in the 30-night OK eyes were much looser than those in the 1-night OK eyes. However, the density and morphology of hemidesmosomes were similar between the OK and RGP eyes at 30 nights of lens wear. CONCLUSIONS: Overnight OK can change the corneal curvature through anterior corneal remodeling, resulting in an effective correction of myopia. However, a decrease in intercellular junctions during long-term wear may compromise the functions of the corneal epithelium as a border mechanical barrier.