Fungal keratitis is a serious blinding eye disease. The development of fungal infections depends primarily on the interaction of fungal virulence with host immune defense factors. The cornea is considered an immune-privileged organ, and resident macrophages are the main immune cells that respond to the heterogeneity exhibited by the microenvironment with their polarization. In the early stage of infection, macrophages polarize towards M1, which promotes inflammation and facilitates fungal clearance but produces a cellular storm that exacerbates immune damage; in the late stage of infection, macrophages polarize towards M2, which suppresses the inflammatory response and facilitates tissue repair, but may be immunosuppressed or even immune escape to the detriment of pathogen clearance. The balance between pro-inflammatory and anti-inflammatory responses is key to maintaining the functional integrity of the cornea. Current antifungal drug therapy is limited, so it is particularly important to find a therapeutic target for the inflammatory response triggered by the immune response in addition to antifungal therapy. In this review, the functional and phenotypic characterization of macrophage subsets associated with fungal keratitis was reviewed, more in-depth research is needed to explore the specific mechanisms by which macrophage polarization and their impact on fungal keratitis. Targeted regulation of macrophage differentiation based on their phenotype and function could be an effective approach to treat and manage fungal keratitis in the future.

Objective To investigate the risk factors associated with prolonged hospitalization in patients diagnosed with diabetic foot ulcers(DFU),to develop a predictive model,and to conduct internal validation of the model.Methods The clinical data of DFU patients admitted to West China Hospital,Sichuan University between January 2012 and December 2022 were retrospectively collected.The subjects were randomly assigned to a training cohort and a validation cohort at a ratio of 7 to 3.Hospital stays longer than 75th percentile were defined as prolonged length-of-stay.A thorough analysis of the risk factors was conducted using the training cohort,which enabled the development of an accurate risk prediction model.To ensure robustness,the model was internally validated using the validation cohort.Results A total of 967 inpatients with DFU were included,among whom 245 patients were identified as having an extended length-of-stay.The training cohort consisted of 622 patients,while the validation cohort comprised 291 patients.Multivariate logistic regression analysis revealed that smoking history(odds ratio[OR]=1.67,95%confidence interval[CI],1.13 to 2.48,P=0.010),Wagner grade 3 or higher(OR=7.13,95%CI,3.68 to 13.83,P＜0.001),midfoot ulcers(OR=1.99,95%CI,1.07 to 3.72,P=0.030),posterior foot ulcers(OR=3.68,95%CI,1.83 to 7.41,P＜0.001),multisite ulcers(OR=2.91,95%CI,1.80 to 4.69,P＜0.001),wound size≥3 cm2(OR=2.00,95%CI,1.28-3.11,P=0.002),and white blood cell count(OR=1.11,95%CI,1.05 to 1.18,P＜0.001)were associated with an increased risk of prolonged length of stay.Additionally,a nomogram was constructed based on the identified risk factors.The areas under the receiver operating characteristic(ROC)curves for both the training cohort and the validation cohort were 0.782(95%CI,0.745 to 0.820)and 0.756(95%CI,0.694 to 0.818),respectively,indicating robust predictive performance.Furthermore,the calibration plot demonstrated optimal concordance between the predicted probabilities and the observed outcomes in both the training and the validation cohorts.Conclusion Smoking history,Wagner grade≥3,midfoot ulcers,posterior foot ulcers,multisite ulcers,ulcer area≥3 cm2,and elevated white blood cell count are identified as independent predictors of prolonged hospitalization.Therefore,it is imperative that clinicians conduct a comprehensive patient evaluation and implement appropriate diagnostic and therapeutic strategies to effectively shorten the length of stay for DFU patients.

OBJECTIVE: To assess the value of single sperm sequencing in preimplantation genetic testing for monogenic disease (PGT-M). METHODS: A Chinese couple with two children whom had died of Spinal muscular atrophy (SMA) and attended the Jiangxi Provincial Maternal and Child Health Care Hospital in June 2020 was selected as the subject. Eleven single sperm samples were isolated by mechanical immobilization and subjected to whole genome amplification. Real-time PCR and Sanger sequencing were used to detect the SMN1 variants in the single sperm samples. Genomic DNA of the wife, her parents and the husband, as well as one single sperm sample harboring the SMN1 variant and two single sperm samples without the variant were used for the linkage analysis. Targeted capture and high-throughput sequencing were carried out to test 100 single nucleotide polymorphisms distributed within 2 Mb up- and downstream the variant site. The haplotypes linked with the SMN1 variants were determined by linkage analysis. Blastocyst embryos were harvested after fertilizing by intracytoplasmic sperm injection. Cells from the trophoblasts of each embryo were biopsied and subjected to whole genome amplification and targeted capture and high-throughput sequencing to determine their carrier status. Chromosomal aneuploidy of wild-type embryos was excluded. An euploid embryo of high quality was transferred. Amniotic fluid sample was taken at 18 weeks of gestation to confirm the status of the fetus. RESULTS: Genetic testing showed that the couple both had deletion of exons 7 ~ 8 of the SMN1 gene. The wife has inherited the deletion from her father, while the husband was de novo. The haplotypes of the husband were successfully constructed by single sperm sequencing. Preimplantation genetic testing has indicated that 5 embryos had harbored the heterozygous variant, 4 embryos were of the wild type, among which 3 were euploid. Prenatal diagnosis during the second trimester of pregnancy has confirmed that the fetus did not carry the deletion. CONCLUSION By single sperm sequencing and PGT-M, the birth of further affected child has been successfully avoided.
Humans ; Pregnancy ; Female ; Child ; Male ; Preimplantation Diagnosis ; East Asian People ; Semen ; Genetic Testing ; Muscular Atrophy, Spinal/genetics* ; Aneuploidy ; Blastocyst/pathology* ; High-Throughput Nucleotide Sequencing ; Spermatozoa

Objective To explore the characteristics of baseline inflammatory markers in diabetic foot patients and their relationship with the prognosis of diabetic foot ulcers.Methods The clinical data of diabetic foot patients(n=495)admitted to West China Hospital,Sichuan University since 2016 were retrospectively collected through the hospital electronic medical record system to analyze the characteristics of inflammatory markers and their relationship with the prognosis of diabetic foot ulcers.Results White blood cell count(WBC),erythrocyte sedimentation rate(ESR),C-reactive protein(CRP),procalcitonin(PCT),and interleukin-6(IL-6)levels were significantly higher in patients defined as grade 4 on the Wagner Scale than those in patients defined as grade 0-3 on the Wagner Scale.Neutrophil percentage(NE％)was higher in Wagner grade-4 patients than those in Wagner grade-0 and grade-1 patients and higher in Wagner grade-3 patients than those in Wagner grade-0 patients.NE％,CRP,PCT,and IL-6 levels were positively correlated with the severity of diabetic foot,with the respective odds ratio(OR)at 95％confidence interval(CI)being 1.038(1.019-1.056),1.019(1.012-1.026),8.225(2.015-33.576),and 1.017(1.008-1.025).Using Wagner grade-0 patients as the reference,patients with higher WBC were more likely to progress to Wagner grade 2,3,and 4,with the respective OR(95％CI)values being 1.260(1.096-1.447),1.188(1.041-1.356),and 1.301(1.137-1.490);patients with higher ESR were more likely to progress to Wagner grade 3 and 4,with the respective OR(95％CI)values being 1.030(1.006-1.054)and 1.045(1.019-1.071).Baseline ESR(P=0.008),CRP(P=0.039),and IL-6(P=0.033)levels were lower in patients who had received antibiotics prior to their admission than those in patients who had not received antibiotics before admission.The levels of WBC,NE％,ESR,PCT,and IL-6 were lower in the full recovery group than those in the group of patients who did not respond to treatment.The higher the levels of NE％and IL-6,the worse the prognosis of diabetic foot ulcers became,with the respective OR(95％CI)values being 1.030(1.010-1.051)and 1.008(1.002-1.013).Conclusion The severity of diabetic foot ulcers increased with the rise in baseline levels of inflammatory markers.Elevated baseline NE％and IL-6 levels suggest a poor prognosis.Our findings suggest that early assessment of diabetic foot infection and standardized antibiotic therapy should be implemented to improve the prognosis.

Objective:To investigate the in vitro interaction between antifungals and tacrolimus against Fusarium solani. Methods:According to Clinical and Laboratory Standards Institute(CLSI) M27-Ed4 and M38-A3, 24 strains of Fusarium solani isolated from clinic were used to test the minimal inhibitory concentration (MIC) of the four antifungals.Chessboard microdilution method was used to evaluate the interaction of tacrolimus (FK506) combined with four antifungals in vitro.The study protocol was approved by the Ethics Committee of Hainan Eye Hospital (No.2017-006). Results:The MIC ranges of fluconazole, voriconazole, natamycin and amphotericin B against 24 strains of Fusarium were 64-256 μg/ml, 1-8 μg/ml, 2-8 μg/ml and 1-8 μg/ml, respectively.Tacrolimus combined with fluconazole, voriconazole, natamycin and amphotericin B had synergistic effects on 12.5% (3/24), 4.2% (1/24), 16.7% (4/24) and 29.2% (7/24) Fusarium, respectively.There were no antagonistic effect in this study. Conclusions:Fusarium is sensitive to natamycin in vitro and partly sensitive to amphotericin B and voriconazole.Synergistic effects exist in few strains of Fusarium but no antagonistic effect.

Background Hainan Island is a tropical and subtropical area in China,with a suitable climate for fungal growth and breeding.Keratomycosis occurred frequently in this area.It is essential to analyze the causative genus and investigate the sensitive drugs prior to the clinical treatment of fungal keratitis.Objective This study attempted to analyze the fungal population of fungal corneal ulcer patients in Hainan Island of China and examine the susceptibility of antifungal drugs.Methods A series cases observational study was carried out.One hundred and seventy-four eyes of 174 fungal corneal ulcer patients were enrolled in Hainan Eye Hospital from February 2014 to May 2016 based on the diagnosis criteria in Ophthalmology.The specimens at corneal lesions were collected under the aseptic conditions,and fungus was examined by KOH based smear.Mycological culture was performed with potatodextrose agar and Sabourand's agar.The 82 cultured causative strains were identified to characterize the spectrum of the species associated with keratitis infections in Hainan Province.Susceptibilities of these causative strains to ten antifungals were determined in vitro by slip diffusion method.Results By smear microscopy,fungal hyphae and spores were found in 34 specimens of 126 patients,with the positive rate 27.0％.In 174 specimens of fungal culture,82 strains of fungi were found,with the positive rate 47.1％,and typing results revealed that the isolates were distributed among species in the fusarium (47.5％),non-spore fungus (12.2％),aspergillus (9.8％) and phoma (9.8％).Nystatin was proved to be the most effective drug to 77 strains of antifungal drug susceptibility,followed by amphotericin B and terbinafine,with the susceptibility of 88.3 ％,72.7 ％,and 71.4％,respectively.Conclusions Fusarium species,non-spore fungus,aspergillus,and phoma are common pathogenic fungi of keratomycosis around Hainan Island in China,and nystatin,amphotericin B and terbinafine are efficient in the treatment of the fungal corneal ulcer.

Objective To explore the dynamic change and clinical signiticance of plasma D-damer level in patients with cerebral hemorrhage and acute craniocerebral injury.Methods 50 patients with cerebral hemorrhage and 40 patients with acute craniocerebral injury were selected,The enzyme-linked immunosorbent assay (ELISA) was used to measure plasma D-dimer level in two groups of patients after onset,and the results were compared with 40 healthy controls.Results The levels of plasma D-dimer in the patients with cerebral hemorrhage were 1.59mg/L,2.10mg/L,1.03 mg/L,0.82mg/L at 3 h,6h,12h,2d after onset,which in the patients with acute craniocerebral injury were 1.61mg/L,2.02mg/L,1.01mg/L and 0.67mg/L,respectively.And the plasma D-dimer levels were 0.50mg/L,0.49mg/L,0.47mg/L,0.48mg/L in the control group at 3h,6h,12h and 2d after onset.The levels of plasma D-dimer in the patients with acute craniocerebral injury were significantly higher than those in the control group,and the differences were statistically significant (t =9.35,12.17,4.03,3.05,all P ＜ O.05).At 7d after onset,the D-dimer levels in the cerebral hemorrhage group and acute craniocerebral injury group were 0.53mg/L,0.55mg/L,respectively,which of the control group was 0.47mg/L,there was no statistically significant difference among the three groups(P ＞ 0.05).Conclusion Cerebral hemorrhage patients and acute craniocerebral injury patients have high coagulation and fibrinolytic activity in brief increase trend,dynamic observation of plasma D-dimer level in patients with cerebral hemorrhage and acute craniocerebral injury is helpful to determine courses,condition and evaluate prognosis.

OBJECTIVETo investigate the characteristics of multifocal electroretinograms (mf-ERG) of different phases in diabetic retinopathy (DR) and its clinical significance.

METHODSMultifocal electroretinograms in patients with DR (I - II stage) were tested with VERIS IV system.

RESULTSIn I - II stage, the absolute values of N1, P1 and N2 response densities, and the N1-P1 and P1-N2 response densities were attenuated is a field of about 45 degrees in diameter.

CONCLUSIONAs a new objective and quantitative examination for spatial visual function, multifocal electroretinograms may be valuable in the diagnosis of diabetic retinopathy.

Aged ; Diabetic Retinopathy ; pathology ; physiopathology ; Electroretinography ; methods ; Female ; Humans ; Male ; Middle Aged ; Retina ; pathology ; physiopathology ; Time Factors