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Aim: To evaluate the potential of two trabecular meshwork (TM)-specific promoters, Chitinase 3-like 1 (Ch3L1) and matrix gla protein (MGP), for improving specificity and safety in glaucoma gene therapy based on self-complementary AAV2 (scAAV2) vector technologies., Methods: An scAAV2 vector with C3 transferase (C3) as the reporter gene (scAAV2-C3) was selected. The scAAV2-C3 vectors were driven by Ch3L1 (scAAV2-Ch3L1-C3), MGP (scAAV2-MGP-C3), enhanced MGP (scAAV2-eMGP-C3) and cytomegalovirus (scAAV2-CMV-C3), respectively. The cultured primary human TM cells were treated with each vector at different multiplicities of infections. Changes in cell morphology were observed by phase contrast microscopy. Actin stress fibers and Rho GTPases/Rho-associated protein kinase pathway-related molecules were assessed by immunofluorescence staining, real-time quantitative polymerase chain reaction and Western blot. Each vector was injected intracamerally into the one eye of each rat at low and high doses respectively. In vivo green fluorescence was visualized by a Micron III Retinal Imaging Microscope. Intraocular pressure (IOP) was monitored using a rebound tonometer. Ocular responses were evaluated by slit-lamp microscopy. Ocular histopathology analysis was examined by hematoxylin and eosin staining., Results: In TM cell culture studies, the vector-mediated C3 expression induced morphologic changes, disruption of actin cytoskeleton and reduction of fibronectin expression in TM cells by inhibiting the Rho GTPases/Rho-associated protein kinase signaling pathway. At the same dose, these changes were significant in TM cells treated with scAAV2-CMV-C3 or scAAV2-Ch3L1-C3, but not in cells treated with scAAV2-eMGP-C3 or scAAV2-MGP-C3. At low-injected dose, the IOP was significantly decreased in the scAAV2-Ch3L1-C3-injected eyes but not in scAAV2-MGP-C3-injected and scAAV2-eMGP-C3-injected eyes. At high-injected dose, significant IOP reduction was observed in the scAAV2-eMGP-C3-injected eyes but not in scAAV2-MGP-C3-injected eyes. Similar to scAAV2-CMV-C3, scAAV2-Ch3L1-C3 vector showed efficient transduction both in the TM and corneal endothelium. In anterior segment tissues of scAAV2-eMGP-C3-injected eyes, no obvious morphological changes were found except for the TM. Inflammation was absent., Conclusion: In scAAV2-transduced TM cells, the promoter-driven efficiency of Ch3L1 is close to that of cytomegalovirus, but obviously higher than that of MGP. In the anterior chamber of rat eye, the transgene expression pattern of scAAV2 vector is presumably affected by MGP promoter, but not by Ch3L1 promoter. These findings would provide a useful reference for improvement of specificity and safety in glaucoma gene therapy using scAAV2 vector., (International Journal of Ophthalmology Press.)

Aim: To investigate the causal gene mutation and clinical characteristics for two Chinese families with autosomal dominant congenital coralliform cataract., Methods: Two Chinese pedigrees with congenital cataract were investigated. Routine ophthalmic examinations were performed on all patients and non-affected family members. Peripheral blood samples were collected, and the genomic DNAs were extracted. The coding regions of proband's DNAs were analyzed with cataract gene panel. The identified mutation was amplified by polymerase chain reaction, and automated sequencing was performed in other members of two families to verify whether the mutated gene was co-segregated with the disease., Results: Congenital coralliform cataract was inherited in an autosomal dominant mode in both pedigrees. For each family, more than half of the family members were affected. All patients presented with severe visual impairment after birth as a result of bilateral symmetric coralliform lens opacification. An exact the same defect in the same gene, a heterozygous mutation of c.70C>A (p. P24T) in exon 2 of γD-crystallin gene, was detected in both probands from each family. Sanger sequencing analysis demonstrated that the mutated CRYGD was co-segregated in these two families., Conclusion: A c.70C>A (p. P24T) variant in CRYGD gene was reconfirmed to be the causal gene in two Chinese pedigrees. It is known that mutated CRYGD caused most of the congenital coralliform cataracts, suggesting that the CRYGD gene is associated with coralliform congenital cataract., (International Journal of Ophthalmology Press.)

Objective: To observe the efficacy of acupuncture combined with Heixiaoyao powder for children with cerebral palsy (liver- qi stagnation, spleen-kidney deficiency syndrome) and its effect on serum immune indexes and nerve growth related protein., Methods: A total of 180 children with cerebral palsy were randomly divided into a combined group (60 cases, 2 cases dropped off), an acupuncture group (60 cases, 4 cases dropped off) and a Chinese medication group (60 cases, 5 cases dropped off). On the basis of conventional treatment, the children in the combined group were treated with acupuncture [Baihui (GV 20), Sishencong (EX-HN 1), Shenting (GV 24), Benshen (GB 13), 30 min each time, twice a day] and Heixiaoyao powder; the children in the acupuncture group were treated with acupuncture, and the children in the Chinese medication group were treated with Heixiaoyao powder, the treatment was same with the combined group. All the children were treated for 6 consecutive days and rest for 1 day, totaling for 8 weeks. The Gesell developmental schedules (GDS) and TCM symptom scores were recorded before treatment, after treatment and at 3-month, 6-month and 12-month follow-up visit; the serum immune indexes (IgA, IgG and IgM) and nerve growth related protein [myelin basic protein (MBP), high mobility group box-1 (HMGB 1 ), neuronspecific enolase (NSE)] were detected before and after treatment. The clinical efficacy of each group was evaluated., Results: The total effective rate was 91.4% (53/58) in the combined group, which was higher than 80.4% (45/56) in the acupuncture group and 78.2% (43/55) in the Chinese medication group ( P <0.05). Compared before treatment, the GDS scores in the three groups were increased and the TCM symptom scores were reduced after treatment and at 3-month, 6-month and 12-month follow-up visit ( P <0.05). The GDS score in the combined group was higher than that in the acupuncture group and the Chinese medication group, and the TCM symptom score was lower than that in the acupuncture group and Chinese medication group ( P <0.05). After treatment, the serum levels of IgA, IgG and IgM in the combined group were increased ( P <0.05), and the serum levels of MBP, HMGB l and NSE were decreased ( P <0.05), and the improvements were superior to those in the acupuncture group and the Chinese medication group ( P <0.05)., Conclusion: Acupuncture combined with Heixiaoyao powder could effectively improve the development quotient in children with cerebral palsy (liver- qi stagnation, spleen-kidney deficiency syndrome), regulate the serum immune indexes and nerve growth related protein levels.

Objective: To study whether diffusion weighted imaging (DWI) and apparent diffusion coefficient (ADC) can reflect angiogenesis and the expression of the vascular endothelial growth factor (VEGF) by analyzing the correlation between the features of DWI and angiogenesis in prostate cancer (PCa)., Methods: We studied the clinical and pathological data of 38 patients with histologically proven PCa, who were examined in the supine position with a 1.5T superconductive magnetic scanner (Siemens Sonata) with a pelvic phased array multi-coil. DWI was obtained by echo planar imaging (EPI) sequence. Another 33 cases of benign prostate hyperplasia (BPH) and 15 healthy volunteers were detected for the ADC value in the PCa and BPH tissues and the peripheral zone (PZ). All the PCa samples were examined for microvascular density (MVD) and VEGF., Results: The ADC values of PCa, BPH and PZ were (49.32 +/- 12.68) x 10(-5) mm2/s, (86.73 +/- 26.75) x 10(-5) mm2/s and (126.25 +/- 27.21) x 10(-5) mm2/s, the former lower than the latter two (P < 0.05). The expressions of MVD and VEGF in PCa were higher than in BPH (P < 0.05). The correlation was negative between the ADC value and MVD of PCa (r = -0.510, P < 0.05) , and positive between the expressions of VEGF and MVD (r = 0.481, P < 0.05). The ADC values of the VEGF-positive and -negative groups were (47.27 +/- 9.55) x 10(-5) mm2/s and (55.06 +/- 11.6) x 10(-5) mm2/s (P < 0.05)., Conclusion: The ADC value reflects the angiogenesis in differentiated prostate cancer, and DWI therefore helps to evaluate the biological features of PCa in vivo.