Characterisation of thickness changes in the peripapillary retinal nerve fibre layer in patients with Leber's hereditary optic neuropathy.

BACKGROUND: Limited studies have identified the changes in peripapillary retinal nerve fibre layer (pRNFL) thickness in patients with chronic Leber's hereditary optic neuropathy (LHON) at different stages of the disease. We aimed to characterise the pRNFL thickness changes in patients with LHON having m.11778G>A (MT-ND4) mutation. METHODS: This retrospective cross-sectional study included 221 eyes from patients with LHON (n=145)-classified into seven groups according to disease duration-and 52 eyes from healthy controls (n=26). All subjects underwent pRNFL examinations. pRNFL thickness of the superior, nasal and inferior, and temporal quadrants, and the 360° average were measured. RESULTS: Within 3 months of onset, the temporal pRNFL thickness decreased significantly, whereas the remaining quadrants and the average pRNFL thickness initially increased. The temporal quadrant (p<0.01) and average pRNFL thickness (p=0.02) significantly decreased at 3-6 months. Excluding that in the nasal quadrant (p=0.93), pRNFL thickness significantly decreased in all other quadrants at 6-9 months. At 9-12 months, the average and individual quadrant pRNFL thicknesses continued to decrease. Compared with 12-24 months, the pRNFL thickness was thinner at 24-60 months and >60 months. CONCLUSIONS: The papillomacular bundle was affected first and preferentially in LHON. pRNFL thickness initially increased and then decreased, corresponding to the retinal ganglion cell swelling and apoptosis. pRNFL thinning first occurred in the temporal quadrant, followed by the inferior and superior quadrants, and finally, the nasal quadrant. pRNFL continued to thin slowly in some quadrants even after 60 months.

Factors associated with rapid improvement in visual acuity in patients with Leber's hereditary optic neuropathy after gene therapy.

PURPOSE: To analyse the factors associated with rapid and significant improvement in visual acuity in patients with Leber's hereditary optic neuropathy (LHON) after gene therapy and explain the theory of this improvement. METHODS: We recruited 149 patients with LHON, who underwent gene therapy, and divided them into two groups according to the absence or presence of rapid and significant visual acuity improvements within 3 days of treatment. A bivariate logistic regression model was used to analyse relevant factors including age, the period between onset and treatment, baseline values of best corrected visual acuity (BCVA), visual field index (VFI) and pretreatment average retinal nerve fibre-layer thickness (GRNFL). The corresponding parameters for the injected and non-injected eyes were analysed separately. RESULTS: The period between onset and treatment, and pretreatment baseline BCVA was significantly associated with rapid and significant improvement in visual acuity for both, the injected and non-injected eyes. Moreover, pretreatment baseline VFI and GRNFL were not significantly associated with rapid and significant improvement in visual acuity. Age was significantly associated with rapid and significant improvement in visual acuity of the injected eyes. CONCLUSION: The period between onset and treatment, and pretreatment baseline BCVA may be important predictors of rapid and significant improvement in visual acuity in patients with LHON after gene therapy.

Dividing CKD stage 3 into G3a and G3b could better predict the prognosis of IgA nephropathy.

Chronic kidney disease (CKD) stage 3 was divided into stage G3a and stage G3b in the 2013 Kidney Disease Improving Global Outcomes guidelines. Whether it is appropriate to regard 45 mL/min/per 1.73 m2 as the threshold value of G3a/G3b staging and whether dividing CKD stage 3 into G3a/G3b plays a useful role in assessing the prognosis of patients with IgA nephropathy (IgAN) remain unknown. Three hundred and ninety patients from the First Affiliated Hospital of Zhengzhou University and Peking University First Hospital diagnosed with IgAN in CKD stage 3 were enrolled and successfully followed up. Cox proportional hazards model was used to analyze hazard ratios of reaching the composite endpoints (doubling of serum creatinine, end-stage renal disease: estimated glomerular filtration rate (eGFR) <15 ml/min/per 1.73 m2 or renal replacement therapy, or death) for patients with different eGFR and risk factors affecting composite endpoints. The Kaplan-Meier method was used to calculate the cumulative renal survival rate of patients. When eGFR was lower than 45 ml/min/per 1.73 m2, the hazard ratio increased sharply for patients in CKD stage 3 who reached the composite endpoints. Renal injury and prognosis were significantly different between patients in the G3a and G3b groups. Stage G3b was a major risk factor affecting prognosis. A threshold value of 45 ml/min/per 1.73 m2 appears appropriate to assess the prognosis of IgAN patients with CKD stage 3. Dividing IgAN patients with CKD stage 3 into G3a and G3b is very useful to help understand disease conditions and for predicting the risk for disease progression.

[Differentiation of mesenchymal stem cells induced by co-culture method].

OBJECTIVE: To investigate the potential differentiation of human mesenchymal stem cells(MSCs)into epithelium-like cells by an in vitro co-culture method. METHODS: The human conjunctival epithelium was obtained by digestion with dispase2, and the MSCs were isolated by density gradient centrifugalization. All cells were identified according to their morphologies and cell-surface antigen profiles by immunocytochemical analysis. The MSCs underwent co-culture with conjunctival epithelium in the manner without cell-to-cell contract. The morphological characterizes of cells were observed under contrast microscope, and the cytokeratin-4 expressions of the differentiated cells were identified by immunocytochemistry staining, reverse transcriptase polymerase chain reaction(RT-PCR), and Western blotting. RESULTS: Immunocytochemistry showed that positive expression of CD29 and negative expression of CD34 in the in vitro cultured MSCs. Cytokeratins4(CK4)was positively expressed in the human conjunctival epithelium. After co-cultured with conjunctival epithelial for 10 days, CK4 was detected in differentiated cells by immunocytochemistry, RT-PCR, and Western blotting. CONCLUSION: MSCs can differentiate into epithe1ium-like cells after having been co-cultured with human conjunctival epithelium.

[Inhibitory and lethal effects of human-mouse chimeric antibody against CD80 on the growth of B lymphoma cell lines Raji and Daudi].

AIM: To study the inhibitory and lethal effects of human-mouse chimeric antibody against CD80 (named ch-4E5) on the growth of B lymphoma cell lines Daudi and Raji. METHODS: Immunofluorescence and flow cytometry were used to analyze the detection of membrane CD80 in Raji and Daudi by ch-4E5. After the co-culture of ch-4E5 with Raji and Daudi, respectively, at the final concentration of 10 mg/L, the expression of Fas and FasL was observated to be at the 0 h, 4 h, 10 h, 16 h, 24 h and 48 h by direct immunofluorescence and flow cytometry, and the blocking effect on cell growth of ch-4E5 was determined at 72 h by MTT assay. MTT assay was also used to study the ADCC effect with PBMC as effector cells and Raji and Daudi as target cells. The efficiency target ratio was 20:1. RESULTS: The combination rate between ch-4E5 and Raji and Daudi was 98.6% and 96.4%, respectively. After the co-culture of ch-4E5 with Raji and Daudi cells for 4 hs, the expression of FasL in Raji began to up-regulate. It reached the peak at 16 h and its positive rate was 16.8%. Compared with human IgG1 control group, it was increased obviously (P<0.01). The expression of Fas increased at 10 h, and then reached the top at 24 h. The combination rate was 15.6%. There was significant deviation compared with human IgG1 control group (P<0.01). Moreover, the expression of FasL and Fas on Daudi was also altered. The trend was similar to these on Raji, and the highest expression was 15.9% and 13.7%, respectively. The inhibitory rate was 34.60% and 32.64% respectively when ch-4E5 with Raji and Daudi had been co-cultured for 72 h (P<0.01). Furthermore, ch-4E5 could mediate the ADCC effect and the maximum killing rate was 55.61% and 54.42%, respectively(P<0.01). CONCLUSION: The human-mouse chimeric antibody against CD80 can inhibit the proliferation of B lymphoma cell lines Daudi and Raji cells through Fab and Fc sections in vitro.