Novel MIP gene mutation causes autosomal-dominant congenital cataract.

AIM: To identify disease-causative mutations in families with congenital cataract. METHODS: Two Chinese families with autosomal-dominant congenital cataract (ADCC) were recruited and underwent comprehensive eye examinations. Gene panel next-generation sequencing of common pathogenic genes of congenital cataract was performed in the proband of each family. Sanger sequencing was used to valid the candidate gene mutations and sequence the other family members for co-segregation analysis. The effect of sequence changes on protein structure and function was predicted through bioinformatics analysis. Major intrinsic protein (MIP)-wildtype and MIP-G29R plasmids were constructed and microinjected into zebrafish single-cell stage embryos. Zebrafish embryonic lens phenotypes were screened using confocal microscopy. RESULTS: A novel heterozygous mutation (c.85G>A; p.G29R) in the MIP gene was identified in the proband of one family. A known heterozygous mutation (c.97C>T; p.R33C; rs864309693) in MIP was found in the proband of another family. In-silico prediction indicated that the novel mutation might affect the MIP protein function. Zebrafish embryonic lens was uniformly transparent in both wild-type PCS2+MIP and mutant PCS2+MIP. CONCLUSION: Two missense mutations in the MIP gene in Chinese cataract families are identified, and one of which is novel. These findings expand the genetic spectrum of MIP mutations associated with cataracts. The functional studies suggest that the novel MIP mutation might not be a gain-of-function but a loss-of-function mutation.

Selective Si-C(sp3) bond cleavage of a silyl-bridged amido alkyl ligand in an yttrium complex.

Compared with Si-C(sp2 and sp) bonds bearing neighboring π-bond hyperconjugative interactions, the activation of robust Si-C(sp3) bonds has proved to be a challenge. Herein, two distinct Si-C(sp3) bond cleavages have been realized by rare-earth-mediated and nucleophilic addition of unsaturated substrates. The reactions of TpMe2Y[κ2-(C,N)-CH(SiH2Ph)SiMe2NSiMe3](THF) (1) with CO or CS2 gave two endocyclic Si-C bond cleavage products, TpMe2Y[κ2-(O,N)-OCCH(SiH2Ph)SiMe2NSiMe3](THF) (2) and TpMe2Y[κ2-(S,N)-SSiMe2NSiMe3](THF) (3), respectively. However, 1 reacted with nitriles such as PhCN and p-R'C6H4CH2CN in a 1 : 1 molar ratio to yield the exocyclic Si-C bond products TpMe2Y[κ2-(N,N)-N(SiH2Ph)C(R)CHSiMe2NSiMe3](THF) (R = Ph (4); R = C6H5CH2 (6H); R = p-F-C6H4CH2 (6F); and R = p-MeO-C6H4CH2 (6MeO)), respectively. Moreover, complex 4 can continuously react with an excess of PhCN to form a TpMe2-supported yttrium complex with a novel pendant silylamido-substituted ß-diketiminato ligand, TpMe2Y[κ3-(N,N,N)-N(SiH2Ph)C(Ph)CHC(Ph)N-SiMe2NSiMe3](PhCN) (5).

Cooperative Rare-Earth/Lithium-Mediated Conversion of White Phosphorus.

The rare-earth/lithium cooperative effect on functionalization of white phosphorus has been investigated. The reaction of diazabutadiene-supported yttrium hydride chelated a LiPPh2 molecule (LY ⋅ THF)2 (µ-H)2 [µ-PPh2 (Li)] (1, L=N,N'-di(2,6-diisopropylphenyl)-1,4-diazabutadiene) with P4 gave two novel mixed Y/Li multinuclear polyphosphorus complexes (LY ⋅ THF)2 [cyclo-P3 ]Li(THF)3 (2) and [Li(THF)4 ]+ [(LY ⋅ THF)3 (norborane-P7 )Li(THF)]- (3), accompanied with the elimination of diphosphorus compound Ph2 PPPh2 (4) and H2 . However, the comparative reaction of yttrium hydride (LY ⋅ THF)2 (µ-H)2 with P4 afforded a trinuclear yttrium pyramid-P4 complex (LY ⋅ THF)3 (µ3 -P(PH)3 ) (5). Further investigations show that 5 cannot continuously react with LiPPh2 to form 2 and 3, and LiPPh2 reacted with P4 to form a Zintl-P7 lithium complex (TMEDA⋅Li)3 (Zintl-P7 ) (6) and 4. These results indicated that the cooperation of Y/Li for activation of P4 is a key for the formation of 2 and 3. All new compounds have been characterized by NMR spectroscopy and single-crystal X-ray diffraction studies.

Aptamer Inhibits Tumor Growth by Leveraging Cellular Proteasomal Degradation System to Degrade c-Met in Mice.

Current action mechanisms for aptamer-based therapeutics depend on occupancy-driven pharmacology to mediate protein functions. We report a new mechanism where aptamers leverage cellular proteasomal degradation system to degrade proteins for cancer treatment. A DNA aptamer (hereinafter referred to as c-Met-Ap) binds to the extracellular domain of mesenchymal-epithelial transition factor (c-Met) and selectively induces c-Met phosphorylation at Y1003 and Y1349. The phosphorylation of Y1003 recruits E3 ubiquitin ligase casitas B-lineage lymphoma, causing c-Met ubiquitination and degradation in the proteasome. Furthermore, c-Met-Ap can induce a decrease in the heterodimeric partner proteins of c-Met and the downstream effector proteins in the c-Met signal axis, effectively inhibiting tumor growth in A549 tumor-bearing BALB/c mice. Our study uncovers a novel, actionable mechanism for aptamer therapeutics and opens a new avenue for developing highly efficient anticancer drugs.

LncTRPM2-AS inhibits TRIM21-mediated TRPM2 ubiquitination and prevents autophagy-induced apoptosis of macrophages in asthma.

Long non-coding RNAs (lncRNAs) play a crucial role in macrophage development but little is known about their role in asthma. Here, we investigated the role of lncRNA lncTRPM2-AS in asthma and found that lncTRPM2-AS participates in the promotion of macrophage inflammation. Downregulation of lncTRPM2-AS promoted apoptosis and inhibited proliferation and production of cytokines including IL-1ß, IL-4, IL-6, IL-10, TNF-α, and TGF-ß. RNA-immunoprecipitation and mass spectrometry indicated that the protein TRPM2 interacted with both lncTRPM2-AS and the E3 ubiquitin ligase TRIM21. LncTRPM2-AS silencing enhanced the interaction between TRIM21 and TRPM2, resulting in elevated levels of ubiquitin-related degradation of TRPM2. Mutation analysis indicated that TRPM2 K1218 is a key site for TRIM21-dependent ubiquitination. Downregulation of lncTRPM2-AS significantly decreased intracellular calcium levels by restraining TRPM2 protein expression, which in turn decreased ROS levels and increased autophagy to promote macrophage apoptosis and reduce cytokine production, together inhibiting macrophage inflammation. Taken together, our findings demonstrate that lncTRPM2-AS blocks the ubiquitination of TRPM2 via TRIM21 and inhibits autophagy-induced apoptosis which may contribute to macrophage inflammation in asthma.

Incorporating Differential Gene Expression Analysis with Predictive Biomarkers to Identify Novel Therapeutic Drugs for Fuchs Endothelial Corneal Dystrophy.

PURPOSE: Based on the differential gene expression analysis for predictive biomarkers with RNA-Sequencing data from Fuchs endothelial corneal dystrophy (FECD) patients, we are aiming to evaluate the efficacy of Library of Integrated Network-based Cellular Signatures (LINCS) perturbagen prediction software to identify novel pharmacotherapeutic targets that can revert the pathogenic gene expression signatures and reverse disease phenotype in FECD. METHODS: A publicly available RNA-seq dataset was used to compare corneal endothelial specimens from controls and patients with FECD. Based on the differential gene expression analysis for predictive biomarkers, we evaluated the efficacy of LINCS perturbagen prediction software to identify novel therapeutic targets that can revert the pathogenic gene expression signatures and reverse disease phenotypes in FECD. RESULTS: The RNA-seq dataset of the corneal endothelial cells from FECD patients revealed the differential gene expression signatures of FECD. Many of the differential expressed genes are related to canonical pathways of the FECD pathogenesis, such as extracellular matrix reorganization and immunological response. The expression levels of genes VSIG2, IL18, and ITGB8 were significantly increased in FECD compared with control. Meanwhile, the expression levels of CNGA3, SMOX, and CERS1 were significantly lower in the FECD than in control. We employed LINCS L1000 Characteristic Direction Signature Search Engine (L1000-CDS2) to investigate pathway-based molecular treatment. L1000-CDS2 predicted that small molecule drugs such as histone deacetylase (HDAC) inhibitors might be a potential candidate to reverse the pathological gene expression signature in FECD. CONCLUSIONS: Based on differential gene expression signatures, several candidate drugs have been identified to reverse the disease phenotypes in FECD. Gene expression signature with LINCS small molecule prediction software can discover novel preclinical drug candidates for FECD.

Polymorphisms and Circulating Plasma Protein Levels of Immune Checkpoints (CTLA-4 and PD-1) Are Associated With Posner-Schlossman Syndrome in Southern Chinese.

Cytotoxic T-lymphocyte associated protein 4 (CTLA-4) and programmed cell death 1 (PD-1) are well-known key immune checkpoints that play a crucial dampening effect on regulating T-cell homeostasis and self-tolerance. In this study, we aimed to evaluate the association between immune checkpoints (CTLA-4 and PD-1) and Posner-Schlossman syndrome (PSS) in a southern Chinese population. A total of 137 patients with PSS and 139 healthy controls from a southern Chinese population were recruited. Five single nucleotide polymorphisms (SNPs) of CTLA-4 (rs733618, rs4553808, rs5742909, rs231775, and rs3087243) and five SNPs of PD-1 (rs10204525, rs2227981, rs2227982, rs41386349, and rs36084323) were genotyped by SNaPshot technique. Soluble CTLA-4 (sCTLA-4) and soluble PD-1 (sPD-1) were determined by ELISA and antibody array assay, respectively. The frequencies of T allele at rs733618 and A allele at rs231775 of CTLA-4 were significantly higher in PSS patients than in healthy controls (corrected p (Pc ) = 0.037; Pc = 0.044, respectively). The haplotype frequencies of CACGG haplotype (rs733618-rs4553808-rs5742909-rs231775-rs3087243) of CTLA-4 and TGAGC haplotype (rs10204525-rs2227981-rs2227982-rs41386349-rs36084323) of PD-1 in the PSS group was significantly lower than those in the control group (Pc = 0.015, p = 0.034, respectively). Circulating plasma levels of sCTLA-4 and sPD-1 in PSS patients were significantly higher than those in controls (all p < 0.001). The present study suggests that CTLA-4 and PD-1 genetic polymorphisms are associated with the susceptibility to PSS in a southern Chinese population. The upregulated circulating plasma protein levels of sCTLA-4 and sPD-1 might provide some hints regarding the dysfunction of immune checkpoints in PSS during the active status.

Human leucocyte antigen alleles confer susceptibility and progression to Graves' ophthalmopathy in a Southern Chinese population.

PURPOSE: To evaluate the contributions of human leucocyte antigen (HLA) class I and II genes in the development of Graves' ophthalmopathy (GO) in a Southern Chinese population. METHODS: Eight HLA loci were genotyped and analysed in 272 unrelated patients with Graves' disease (GD) or the proptosis and myogenic phenotypes of GO, and 411 ethnically matched control subjects. RESULTS: The allele frequencies of HLA-DRB1*16:02 and -DQB1*05:02 in the GD, proptosis and myogenic groups, HLA-B*38:02 and -DQA1*01:02 in the myogenic group were significantly higher than those in the control group, respectively (all corrected p values <0.05, OR >2.5). The haplotype frequencies of HLA-DRB1*16:02-DQA1*01:02-DQB1*05:02 and HLA-DRB1*16:02-DQA1*01:02-DQB1*05:02-DPA1*02:02-DPB1*05:01 in the proptosis and myogenic groups, and HLA-A*02:03-B*38:02-C*07:02 and HLA-A*02:03-B*38:02-C*07:02-DRB1*16:02-DQA1*01:02-DQB1*05:02-DPA1*02:02-DPB1*05:01 in the myogenic group were significantly higher than those in the control group respectively (all corrected p values <0.05, OR >2.5). The potential epitopes ('FLGIFNTGL' of TSHR, 'IRHSHALVS', 'ILYIRTNAS' and 'FVFARTMPA' of IGF-1R) were fitted exactly in the peptide-binding groove between HLA-DRA1-DRB1*16:02 heterodimer, and the epitopes ('ILEITDNPY' of THSR, 'NYALVIFEM' and 'NYSFYVLDN' of IGF-1R) were also fitted exactly in the peptide-binding groove between HLA-DQA1*01:02-DQB1*05:02 heterodimer. CONCLUSIONS: The HLA-DRB1*16:02 and -DQB1*01:02 alleles might be risk factors for GD including the proptosis and myogenic phenotypes of GO. The alleles HLA-B*38:02, -DQA1*01:02, the HLA haplotypes consisting of HLA-B*38:02, -DRB1*16:02, -DQA1*01:02 and -DQB1*05:02 might be susceptibility risk factors for GO. Simultaneously, some epitopes of TSHR and IGF-1R tightly binding to groove of HLA-DRA1-DRB1*16:02 or HLA-DQA1*01:02-DQB1*05:02 heterodimers might provide some hints on presenting the pathological antigen in GO.

Frequency of presenting visual acuity and visual impairment in Chinese college students.

AIM: To obtain the baseline data on presenting visual acuity (PVA) and evaluate the prevalence and associated factors for visual impairment based on PVA in 9070 Chinese college students. METHODS: The freshmen at a university in southern China, including 6527 undergraduate students and 2543 graduate students, were investigated for some socio-demographic characteristics and underwent routine medical examination, including measuring PVA, height, and weight. Visual impairment was defined according to the new World Health Organization criteria for blindness and visual impairment. RESULTS: In 9070 college students, the mean PVA in the better eye was 0.094±0.163 logMAR. The prevalence of visual impairment based on PVA was 2.7%. Only 38.3% college students had normal visual acuity [PVA equal to 0 logMAR (20/20) in both eyes]. There were 69.8% of students wearing spectacles. Logistic regression showed that home region (non-Guangdong provinces, P<0.0001, OR=1.70) was risk factor for visual impairment while BMI (P=0.001, OR=0.92) was protective factor from visual impairment. Ethnicity (Han Chinese, P<0.0001, OR=3.17) was risk factor for wearing spectacles while age (P=0.01, OR=0.90) was protective factor from wearing spectacles. CONCLUSION: This study provides the baseline data on PVA and the prevalence of visual impairment in Chinese college students. Our analyses reveal that BMI and home region are associated factors for visual impairment based on PVA, while age and ethnicity are associated factors for wearing spectacles.

Lithium chloride confers protection against viral myocarditis via suppression of coxsackievirus B3 virus replication.

Viral myocarditis (VMC) is a type of inflammation affecting myocardial cells caused by viral infection and has been an important cause of dilated cardiomyopathy (DCM) worldwide. Type B3 coxsackievirus (CVB3), a non-enveloped positive-strand RNA virus of the Enterovirus genus, is one of most common agent of viral myocarditis. Till now, effective treatments for VMC are lacking due to lack of drugs or vaccine. Lithium chloride (LiCl) is applied in the clinical management of manic depressive disorders. Accumulating evidence have demonstrated that LiCl, also as an effective antiviral drug, exhibited antiviral effects for specific viruses. However, there are few reports of evaluating LiCl's antiviral effect in mice model. Here, we investigated the inhibitory influence of LiCl on the CVB3 replication in vitro and in vivo and the development of CVB3-induced VMC. We found that LiCl significantly suppressed CVB3 replication in HeLa via inhibiting virus-induced cell apoptosis. Moreover, LiCl treatment in vivo obviously inhibited virus replication within the myocardium and alleviated CVB3-induced acute myocarditis. Collectively, our data demonstrated that LiCl inhibited CVB3 replication and negatively regulated virus-triggered inflammatory responses. Our finding further expands the antiviral targets of LiCl and provides an alternative agent for viral myocarditis.

[Digital simulation of unipedicular thoracolumbar vertebroplasty puncture].

OBJECTIVE: To measure such operative parameters of unipedicular kyphoplasty as optimal entry point, angle and depth so as to provide rationales for its clinical management and formulate a standardized protocol for unipedicular vertebroplasty. METHODS: Ten dry thoracolumbar specimens were prepared for measurement. The entry and target points were defined according to the Roy-Camille method. A 3mm Kirschner wire was used to puncture and view in the anteroposterior and lateral aspects of radiography until a satisfying position. The outside oblique and upward oblique angles were measured on the radiographic pictures. After extraction, the depth of Kirschner wire was measured. The positions of entry point were changed and the largest upward oblique angle and largest declination angle measured on the radiographic pictures. RESULTS: For safe puncturing, as the outside oblique and upward oblique angles enlarged from T(11) to L(3), the length enlarged from T(11) to T(12) and L(1) to L(3). The accepted error was that the largest upward oblique angle and largest declination angle enlarged from T(11) to L(3). The alteration range for outside oblique angle was extremely narrow. CONCLUSION: The experimental results provide the guiding data for the operative management of unipedicular thoracolumbar vertebroplasty. If the pedicle is too small or the angle too narrow, the operative sophistication of vertebroplasty will be highly demanding.