A novel mutation of p.F32I in GJA8 in human dominant congenital cataracts

AIM： To identify a causative mutation in a three-generation family with autosomal dominant congenital total cataract and dissect the molecular consequence of the identified mutation.METHODS： Clinical and ophthalmological examinations were performed on the affected and unaffected family members. Mutation were screened in recruited family members by polymerase chain reaction（PCR） of the two reported genes（CRYAA and GJA8） which were linked to human total cataracts and direct sequencing of the PCR product. The molecular consequences of the identified mutation was dissected. The plasmids carrying wild-type and mutant mouse ORF of Gja8, coding for connexin 50（Cx50）, were generated and ectopic expressed in 293 cells. Recombinant protein expression and cellular localization of recombinated Cx50 were assessed by confocal microscopy.RESULTS： Clinical and ophthalmological examinations were performed on the affected and unaffected family members. Mutation were screened in recruited family members by PCR of the two reported genes（CRYAA and GJA8） which were linked to human total cataractsand direct sequencing of the PCR product. The molecular consequences of the identified mutation was dissected.The plasmids carrying wild-type and mutant mouse ORF of Gja8, coding for Cx50, were generated and ectopic expressed in 293 cells. Recombinant protein expression and cellular localization of recombinated Cx50 were assessed by confocal microscopy. CONCLUSION： This study has identified a novel cataract mutation in GJA8, which adds a novel mutation to the existing spectrum of Cx50 mutations with cataract.The molecular consequences of p.F32 I mutation in GJA8 exclude instability and the mislocalization of mutant Cx50 protein.

A novel mutation of RPGR in a Chinese family with X-linked retinitis pigmentosa

·AIM:To identify potential mutations and elucidate the clinical findings of male patients and female carriers of X-Iinked retinitis pigmentosa(XLRP)in a Chinese family.·METHODS:A four generation pedigree was collected that consisted of 20 individuals.Genomic DNA was extracted from peripheral blood,and then the target fragments were amplified by PCR and sequenced directly.In addition,all affected patients and female carriers underwent comprehensively ophthalmic evaluation.·RESULTS:A novel mutation c.2865 G>A p.W955 X in RPGR gene was identified of this family,including four affected individuals and eight carriers.All male patients,aging from 7 to 31 y,tended to have more various,even potentially deleterious clinical features of RP.At the same time,individuals with heterozygous mutations(carriers)manifested a wide spectrum of clinical features.Herein,only two male patients and three female carriers manifested pathological myopia(PM).Among the female carriers,half of subjects who harbor poor visual acuity suffered esotropia or exotropia.Additionally,16.7%and 66.7%of carriers had abnormal electroretinogram(ERG)and fundus,respectively.·CONCLUSION:In this study,a novel mutation of the RPGR gene is identified,which broadens the spectrum of RPGR mutations,and elaborates the relationship between genotype and phenotype.

Transcriptional and signaling regulation in neural crest stem cell-derived melanocyte development： do all roads lead to Mitf？

人的 neurocristopathies 包括神经的冠(NC ) 的很多症候群，肿瘤，和 dysmorphologies 干细胞衍生物。在最近的年里，许多白看到基因在源于 NC 茎的 neurocristopathies 与 hypopigmentary 混乱和聋被联系了导出房间的 melanocyte 缺乏在开发期间。这些包括 PAX3， SOX10， MITF， SNAI2， EDNRB， EDN3，工具包，和 KITL。最近的研究在 melanocyte 开发在交往的基因的复杂网络揭示了令人吃惊的新卓见进 MITF 的一个中央角色。在这个观点，我们提供一些当前的调查结果的概述并且在 NC 茎期间探索这些基因的复杂功能的角色导出房间的 melanocyte 开发。

Novel mutations in PDE6B causing human retinitis pigmentosa

AIM: To identify the genetic defects of a Chinese patient with sporadic retinitis pigmentosa (RP). METHODS: Ophthalmologic examinations were performed on the sporadic RP patient, 144 genes associated with retinal diseases were scanned with capture next generation sequencing (CNGS) approach. Two heterozygous mutations in PDE6B were confirmed in the pedigree by Sanger sequencing subsequently. The carrier frequency of PDE6B mutations of reported PDE5B mutations based on the available two public exome databases (1000 Genomes Project and ESP6500 Genomes Project) and one in-house exome database was investigated. RESULTS: We identified compound heterozygosity of two novel nonsense mutations c.1133G>A (p.W378X) and c.2395C>T (p.R799X) in PDE6B, one reported causative gene for RP. Neither of the two mutations in our study was presented in three exome databases. Two mutations (p.R74C and p.T6041) in PDE5B have relatively high frequencies in the ESP6500 and in-house databases, respectively, while no common dominant mutation in each of the database or across all databases. CONCLUSION: We demonstrates that compound heterozygosity of two novel nonsense mutations in PDE6B could lead to RP. These results collectively point to enormous potential of next-generation sequencing in determining the genetic etiology of RP and how various mutations in PDE9B contribute to the genetic heterogeneity of RP.

R102W mutation in the RS1 gene responsible for retinoschisis and recurrent glaucoma

AIM: To identify the mutations in RS1 gene associated with typical phenotype of X-linked juvenile retinoschisis(XLRS) and a rare condition of concomitant glaucoma. ·METHODS: Complete ophthalmic examinations were performed in the proband. The coding regions of the RS1 gene that encode retinoschisin were amplified by polymerase chain reaction and directly sequenced. ·RESULTS: The proband showed a typical phenotype of XLRS with large peripheral retinal schisis in both eyes,involving the macula and combined with foveal cystic change,reducing visual acuity. A typical phenotype of recurrent glaucoma with high intraocular pressure(IOP) and reduced visual field was also demonstrated with the patient. Mutation analysis of RS1 gene revealed R102W(c.304C】T) mutations in the affected male,and his mother was proved to be a carrier with the causative mutation and another synonymous polymorphism(c.576C】CT). ·CONCLUSION: We identified the genetic variations of a Chinese family with typical phenotype of XLRS and glaucoma. The severe XLRS phenotypes associated with R102W mutations reveal that the mutation determines a notable alteration in the function of the retinoschisin protein. Identification of the disease-causing mutation is beneficial for future clinical references.

A Light-Deprivation Mouse Model Potentially for Studying the Complete Congenital Stationary Night Blindness

Current rodent models of the complete congenital stationary night blindness (CSNB1) were time- consuming in breeding and validation, which makes it imperative to find a more “easily handle” animal model to broaden our understanding of this disorder. In the present study, a light-deprivation (LD) mouse model was made to validate whether it was a more “suitable” animal mode for investigating the pathogenesis of the CSNB1. Compared with controls, the LD mice exhibited a remarkable reduction in the amplitude of the dark-adapted electroretinogram (ERG) b-wave, the Max-ERG b-wave and also the oscillatory potentials (Ops), indicating an abnormal activity of rod bipolar cells in the retina. However, the ERG a-wave was relatively normal in the LD mice, which was quite consistent with what was confirmed in previously reported animal models of the CSNB1 and CSNB patients. Taken together, the LD mouse model showed CSNB1-like negative ERG responses as evidenced by the abnormal b-wave. Our study will provide a potentially useful animal model to decipher the pathogenesis of the CSNB1.

A Short Glance at the Neural Circuitry Mechanism Underlying Depression

Depression leads to a large social burden because of its substantial impairment and disability in everyday activities. The prevalence and considerable impact of this disorder call for a better understanding of its pathophysiology to improve the diagnosis, treatment and prevention. Though productive animal models and pathophysiological theories have been documented, it is still very far to uncover the complex array of symptoms caused by depression. Moreover, the neural circuitry mechanism underlying behavioral changes in some depression-like behavior animals is still limited. Changes in the neural circuitry of amygdala, dorsal raphe nucleus, ventral tegmental area, hippocampus, locus coeruleus and nucleus accumbens are related to depression. However, the interactions between individual neural circuitry of different brain areas, have not yet been fully elucidated. The purpose of the present review is to examine and summarize the current evidence for the pathophysiological mechanism of depression, with a focus on the neural circuitry, and emphasize the necessity and importance of integrating individual neural circuitry in different brain regions to understand depression.

The precision and agreement of corneal thickness and keratometry measurements with SS-OCT versus Scheimpflug imaging

Purpose:To assess the repeatability and reproducibility of swept-source optical coherence tomography(SS-OCT)and Scheimpflug system and evaluate the agreement between the two systems in measuring multiple corneal regions in children.Methods:Pachymetric and keratometric maps for both systems were evaluated.Central,midperipheral and peripheral corneal thickness(CT),keratometry and astigmatism power vectors were recorded.The three outcomes yielded by the same observer were used to assess intraobserver repeatability.The differences in the mean values provided by each observer were used to evaluate interobserver reproducibility.Within-subject standard deviation,test-retest repeatability(TRT)and coefficient of variation(CoV)were used to analyze the intraobserver repeatability and interobserver reproducibility.Paired T-test and Bland-Altman were used to appraise interdevice agreement.Results:Seventy-eight eyes of 78 children were included.The CoV was≤2.12 and 1.10%,respectively,for repeatability and reproducibility.TRT and CoV were lower for central and paracentral CT measurements than for peripheral measurements.The SS-OCT device generated higher precision when acquiring CT data,whereas Scheimpflug system showed higher reliability when measuring corneal keratometry.Although the CT readings measured using SS-OCT were significantly thinner than Scheimpflug device(P<0.001),the central and thinnest CT values were still of high agreement.The interdevice agreement of keratometry measurement was high for the central corneal region and moderate for the paracentral and peripheral areas.Conclusions:The precision of CT measurements by SS-OCT was higher,while the reliability of keratometry measurements by the Scheimpflug system was higher in children.Apart from the measured values in the central corneal region,the thickness and keratometry readings should not be considered interchangeable between the two systems.

Effect of orthokeratology on precision and agreement assessment of a new sweptsource optical coherence tomography biometer

Background:To evaluate the effect of orthokeratology on precision of measurements in children using a new swept-source optical coherence tomography(SS-OCT)optical biometer(OA-2000),and agreement between its measurements and those provided by the commonly used IOLMaster based on partial coherence interferometry(PCI).Methods:This study recruited fifty-one eyes of 51 normal children(8–16 years).An operator took measurements with the two biometers.Then,a second operator took measurements with the SS-OCT biometer.After orthokeratology was performed for one month,the same operators repeated the same procedures.Axial length(AL),mean keratometry(Km)at 2.5 mm and 3.0 mm diameters(Km2.5 and Km3.0),central corneal thickness(CCT),anterior chamber depth(ACD),lens thickness(LT)and corneal diameter(CD)were analyzed.Results:With the SS-OCT optical biometer,the test-retest repeatability of AL measurements was<0.06 mm.For all parameters,the coefficients of variation were<1.23%and the intraclass correlation coefficients were>0.95.The 95%limits of agreement of difference between the two devices for CD parameter were up to 1.53 mm.After orthokeratology,the fluctuation ranges of difference for Km3.0 measurement was 1.11 times higher than before orthokeratology,while the absolute values of difference for AL,Km2.5,ACD and CD measurements were comparable.Conclusions:Before and after orthokeratology,the SS-OCT biometer showed high repeatability and reproducibility for all measurements.Wearing orthokeratology contact lenses affected the agreement between SS-OCT and PCI biometers for Km3.0 measurements.The CD measurement showed poor agreement between the two devices.

Structural insights into the CRISPR-Cas-associated ribonuclease activity of Staphylococcus epidermidis Csm3 and Csm6: a step for the development of novel genome editing tool

CRISPR-Cas systems has been developed as genome editing tools,which are originally discovered as sequence-specific defense system utilized by bacteria and archaea to fight with the invasion by foreign nucleic acids such as phages or plasmids.According to the current classification groups of CRISPR-Cas system,Type I,Type III and Type IV comprise the Class 1 systems for their function through multisubunit Cas protein complex to interference the targeted nucleic acids.In contrast,Class 2 systems,consisting of

The precision and agreement of corneal thickness and keratometry measurements with SS-OCT versus Scheimpflug imaging

Purpose:To assess the repeatability and reproducibility of swept-source optical coherence tomography(SS-OCT)and Scheimpflug system and evaluate the agreement between the two systems in measuring multiple corneal regions in children.Methods:Pachymetric and keratometric maps for both systems were evaluated.Central,midperipheral and peripheral corneal thickness(CT),keratometry and astigmatism power vectors were recorded.The three outcomes yielded by the same observer were used to assess intraobserver repeatability.The differences in the mean values provided by each observer were used to evaluate interobserver reproducibility.Within-subject standard deviation,test-retest repeatability(TRT)and coefficient of variation(CoV)were used to analyze the intraobserver repeatability and interobserver reproducibility.Paired T-test and Bland-Altman were used to appraise interdevice agreement.Results:Seventy-eight eyes of 78 children were included.The CoV was≤2.12 and 1.10%,respectively,for repeatability and reproducibility.TRT and CoV were lower for central and paracentral CT measurements than for peripheral measurements.The SS-OCT device generated higher precision when acquiring CT data,whereas Scheimpflug system showed higher reliability when measuring corneal keratometry.Although the CT readings measured using SS-OCT were significantly thinner than Scheimpflug device(P<0.001),the central and thinnest CT values were still of high agreement.The interdevice agreement of keratometry measurement was high for the central corneal region and moderate for the paracentral and peripheral areas.Conclusions:The precision of CT measurements by SS-OCT was higher,while the reliability of keratometry measurements by the Scheimpflug system was higher in children.Apart from the measured values in the central corneal region,the thickness and keratometry readings should not be considered interchangeable between the two systems.

Progressive cracking technique for phacoemulsification of superhard cataracts: a case report

Background:Complete nuclear disassembly of superhard cataracts cannot always be achieved by phaco chop,which is considered one of the best techniques for dealing with hard cataracts.We present a phaco chopprogressive cracking technique to divide superhard cataracts completely.Case presentation:We presented a case of cataract with over Grade V nucleus sclerosis and very low density of corneal endothelial cell(812 cells/mm^(2)).By performing the cataract surgery with our phaco chop-progressive cracking technique,the corneal endothelial cells were well protected and the patient’s visual acuity was markedly improved from finger counting at 40 cm to 20/200 the day after surgery without obvious corneal edema.Conclusions:Although an initial learning curve was needed,this phaco chop-progressive cracking technique could be of particular benefit to the superhard cataract,especially in patients with low density of corneal endothelial cells.

Dr. Jian Zhou： The great inventor of cervical ：ancer vaccine

In November 2005, scientists gathered at Times Square in New York City announced the exciting news of the successful development of the world＇s first cervical cancer vaccine. The cervical cancer vaccine, which is able to protect unexposed women against the infection with four HPV strains responsible for 70% of cervical cancers, was approved for human use by the US Federal Drug Administration （FDA） and a number of other countries.

Sirius Scheimpflug-Placido versus. ultrasound pachymetry for central corneal thickness: meta-analysis

Background:To compare the difference in central corneal thickness(CCT)measurements in normal eyes between a rotating Scheimpflug camera combined with a Placido-disk corneal topographer(Sirius,CSO,Italy)and ultrasound pachymetry(USP).Methods:A systematic literature search was conducted for relevant studies published on PubMed,Medline,EMBASE,and the Cochrane Library and ClinicalTrials.gov from inception to August 1st,2019.Primary outcome measures were CCT measurements between Sirius and USP.A random effects model was used to pool CCT measurements.Results:A total of twelve studies involving 862 eyes were included in this meta-analysis.The meta-analysis found CCT measurements between Sirius and USP to be statistically significantly different(P<0.0001).The mean difference between Sirius and USP was-11.26μm with a 95%confidence interval(CI)(-16.92μm,-5.60μm).The heterogeneity Was IP=60%(P=0.004).Conclusion:CCT measurements with the Sirius Scheimpflug-Placido topographer were statistically significantly lower than USP.However,it may be argued that the mean difference of 11.26μm is not a clinically significant difference.

Sirius Scheimpflug–Placido versus ultrasound pachymetry for central corneal thickness:meta-analysis

Background:To compare the difference in central corneal thickness(CCT)measurements in normal eyes between a rotating Scheimpflug camera combined with a Placido-disk corneal topographer(Sirius,CSO,Italy)and ultrasound pachymetry(USP).Methods:A systematic literature search was conducted for relevant studies published on PubMed,Medline,EMBASE,and the Cochrane Library and ClinicalTrials.gov from inception to August 1st,2019.Primary outcome measures were CCT measurements between Sirius and USP.A random effects model was used to pool CCT measurements.Results:A total of twelve studies involving 862 eyes were included in this meta-analysis.The meta-analysis found CCT measurements between Sirius and USP to be statistically significantly different(P<0.0001).The mean difference between Sirius and USP was−11.26μm with a 95%confidence interval(CI)(−16.92μm,−5.60μm).The heterogeneity was I^(2)=60%(P=0.004).Conclusion:CCT measurements with the Sirius Scheimpflug-Placido topographer were statistically significantly lower than USP.However,it may be argued that the mean difference of 11.26μm is not a clinically significant difference.

Keratolenticular adhesion removal for type 2 Peters anomaly:a case report

Background:Type 2 Peters anomaly is a rare anterior segment disorder characterized by central corneal leukoma with keratolenticular adhesion and cataract.Performing cataract surgery without corneal tissue transplantation in patients of type 2 Peters anomaly is extremely rare and challenging.We present a case of type 2 Peters anomaly treated by peeling off the adhesion without penetrating keratoplasty(PKP),in which restoration of corneal transparency is observed.Case presentation:An 11-month-old female infant of type 2 Peters anomaly presented with bilateral corneal opacity with distinct demarcation,keratolenticular adhesion and cataract,which was first noted at the age of 3 months.By peeling off the adhesion from corneal endothelium combined with lensectomy and vitrectomy,there was a gradual reduction in corneal opacity and improvement in visual acuity after surgery over a 2-year period.Her visual acuity had improved from light perception preoperatively to 20/50 at the latest follow-up.No sight-threatening postoperative complications were noted.Conclusion:It is safe and effective to peel off the keratolenticular adhesion in patients of type 2 Peters anomaly presented with distinctly demarcated corneal opacity.

Keratolenticular adhesion removal for type 2 Peters anomaly:a case report

Background:Type 2 Peters ano maly is a rare anted or segment disorder characterized by central corneal leukoma with keratolenticular adhesion and cataract.Performing cataract surgery without corneal tissue transplantation in patients of type 2 Peters anomaly is extremely rare and challenging.We present a case of type 2 Peters anomaly treated by peeling off the adhesion without penetrating keratoplasty(PKP),in which restoration of corneal transparency is observed.Case presentation:An 11-month-old female infant of type 2 Peters anomaly presented with bilateral corneal opacity with distinct demarcation,keratolenticular adhesion and cataract,which was first noted at the age of 3 months.By peeling off the adhesion from comeal endothelium combined with lensectomy and vitrectomy,there was a gradual reduction in corneal opacity and improvement in visual acuity after surgery over a 2-year period.The visual acuity had improved from light perception preoperatively to 20/50 at the latest follow-up.No sight-threatening postoperative complications were noted.Conclusion:It is safe and effective to peel off the keratolenticular adhesion in patients of type 2 Peters anomaly presented with distinctly demarcated corneal opacity.

Functional non-homologous end joining patterns triggered by CRISPR/Cas9 in human cells

CRISPR/Cas9-mediated genome engineering technologies are now widely applied in various organisms,including mouse and human cells（Cong et al.,2013;Mali et al.,2013;Yang et al.,2013;Hsu et al.,2014）.The most widely used customized CRISPR/Cas9（Sp Cas9）is derived from Streptococcus pyogenes（Cong et al.,2013）.

Can SpRY recognize any PAM in human cells?

The application of clustered regularly interspaced short palindromic repeats(CRISPR)and CRISPR-associated proteins(Cas)can be limited due to a lack of compatible protospacer adjacent motif(PAM)sequences in the DNA regions of interest.Recently,SpRY,a variant of Streptococcus pyogenes Cas9(SpCas9),was reported,which nearly completely fulfils the PAM requirement.Meanwhile,PAMs for Sp RY have not been well addressed.In our previous study,we developed the PAM Definition by Observable Sequence Excision(PAM-DOSE)and green fluorescent protein(GFP)-reporter systems to study PAMs in human cells.Herein,we endeavored to identify the PAMs of SpRY with these two methods.The results indicated that 5’-NRN-3’,5’-NTA-3’,and 5’-NCK-3’could be considered as canonical PAMs.5’-NCA-3’and 5’-NTK-3’may serve as non-priority PAMs.At the same time,PAM of 5’-NYC-3’is not recommended for human cells.These findings provide further insights into the application of SpRY for human genome editing.