Free Radical Enhancer Xenobiotic is an Inducer of Cataract in Rabbit

Free radical enhancers, diquat, paraquat, plumbagin and juglone were used to study the oxy radical-induced damage to the rabbit lens in vitro and in vivo. Each compound caused a 6–8 fold increase in malondialdehyde (MDA) and a 30–55% decrease in reduced glutathione of the lens in vim. These peroxidative and oxidative changes were potentiated in the presence of 100% 0., abolished by N, and prevented by desferal-Mn (III) (DF-Mn) or liposomal superoxide dismutase (LSOD) indicating the involvement of O₂^?.

Diquat injected intravitreally as a single dose (300nmole in 30μl of isotonic saline) in the right eye of a 5-wk-old Dutch belted rabbit, induced early cataract after 24–72h. The lens of the contralateral control eye injected with isotonic saline had no change. In the right eye, O₂,^? and OH -productions were significantly (P < 0.01) higher; O₂-, was about 16 fold higher in the aqueous humor and vitreous humor, and 5 fold in the lens and retina, and OH. was 35 fold higher in the aqueous humor, 2 fold in vitreous humor and 5 fold in the lens and retina as compared to the respective tissues of the control eye. Enhanced lipid peroxidation in the lens was apparent from the higher levels of MDA and formation of aminophospholipid-MDA Schiff-base conjugates.

We propose that cyclic oxidation-reduction of xenobiotics coupled to the endogenous redox systems in the eye, could generate oxy radicals in excessive amounts, triggering cataractogcnesis.     

Site‐specific rapid deamidation and isomerization in human lens αA‐crystallin in vitro

Recent studies have suggested that the isomerization/racemization of aspartate residues in proteins increases in aged tissues. One such residue is Asp151 in lens‐specific αA‐crystallin. Although many isomerization/racemization sites have been reported in various proteins, the factors that lead to those modifications in proteins in vivo remain obscure. Therefore, an in vitro system is needed to assess the mechanisms of modifications of Asp under various conditions. Deamidation of Asn to Asp in proteins occurs more rapidly than isomerization/racemization of Asp, although the reaction passes through the same intermediate in both pathways. Here, therefore, we replaced Asp151 in human lens αA‐crystallin with Asn by using site‐directed mutagenesis. The recombinant protein was expressed in Escherichia coli and used to investigate the deamidation/isomerization/racemization of Asn151 after incubation at 50°C for various durations and under different pH. After incubation, the mutant αA‐crystallin was subjected to enzymatic digestion followed by liquid chromatography–MS/MS to evaluate the ratio of modifications in Asn151‐containing peptides. The Asp151Asn αA‐crystallin mutant showed rapid deamidation to Asp with the formation of specific Asp isomers. In particular, deamidation increased greatly under basic conditions. By contrast, subunit–subunit interactions between αA‐crystallin and αB‐crystallin had little effect on the modification of Asn151. Our findings suggest that the Asp151Asn αA‐crystallin mutant represents a good in vitro model protein to assess deamidation, isomerization, and the racemization intermediates. Furthermore, our in vitro results show a different trend from in vivo data, implying the presence of specific factors that induce racemization from L‐Asp to D‐Asp residues in vivo.     

Nucleotide-Specific Recognition of Iron-Responsive Elements by Iron Regulatory Protein 1

IRP1 [iron regulatory protein (IRP) 1] is a bifunctional protein with mutually exclusive end-states. In one mode of operation, IRP1 binds iron-responsive element (IRE) stem–loops in messenger RNAs encoding proteins of iron metabolism to control their rate of translation. In its other mode, IRP1 serves as cytoplasmic aconitase to correlate iron availability with the energy and oxidative stress status of the cell. IRP1/IRE binding occurs through two separate interfaces, which together contribute about two-dozen hydrogen bonds. Five amino acids make base-specific contacts and are expected to contribute significantly to binding affinity and specificity of this protein:RNA interaction. In this mutagenesis study, each of the five base-specific amino acids was changed to alter binding at each site. Analysis of IRE binding affinity and translational repression activity of the resulting IRP1 mutants showed that four of the five contact points contribute uniquely to the overall binding affinity of the IRP1:IRE interaction, while one site was found to be unimportant. The stronger-than-expected effect on binding affinity of mutations at Lys379 and Ser681, residues that make contact with the conserved nucleotides G16 and C8, respectively, identified them as particularly critical for providing specificity and stability to IRP1:IRE complex formation. We also show that even though the base-specific RNA-binding residues are not part of the aconitase active site, their substitutions can affect the aconitase activity of holo-IRP1, positively or negatively.     

Role of long non‐coding RNA MIAT in proliferation,apoptosis and migration of lens epithelial cells: a clinical and in vitro study

Age‐related cataract is among the most common chronic disorders of ageing and is the world's leading blinding disorder. Long non‐coding RNAs play important roles in several biological processes and complicated diseases. However, the role of lncRNAs in the setting of cataract is still unknown. Here, we extracted total RNAs from the transparent and age‐matched cataractous human lenses, and determined lncRNA expression profiles using microarray analysis. We found that 38 lncRNAs were differentially expressed between transparent and cataractous lenses. 17 of 20 differentially expressed lncRNAs were further verified by quantitative RT‐PCRs. One top abundant lncRNA, MIAT, was specifically up‐regulated both in the plasma fraction of whole blood and aqueous humor of cataract patients. MIAT knockdown could affect the proliferation, apoptosis and migration of Human lens epithelial cells (HLECs) upon oxidative stress. Posterior capsule opacification (PCO) is a common complication of cataract surgery, which is associated with abnormal production of inflammatory factors. MIAT knockdown could repress tumour necrosis factor‐α‐induced abnormal proliferation and migration of HLECs, suggesting a potential role of MIAT in PCO‐related pathological process. Moreover, we found that MIAT acted as a ceRNA, and formed a feedback loop with Akt and miR‐150‐5p to regulate HLEC function. Collectively, this study provides a novel insight into the pathogenesis of age‐related cataract.     

Within‐host interactions shape virulence‐related traits of trematode genotypes

Within‐host interactions between co‐infecting parasites can significantly influence the evolution of key parasite traits, such as virulence (pathogenicity of infection). The type of interaction is expected to predict the direction of selection, with antagonistic interactions favouring more virulent genotypes and synergistic interactions less virulent genotypes. Recently, it has been suggested that virulence can further be affected by the genetic identity of co‐infecting partners (G × G interactions), complicating predictions on disease dynamics. Here, we used a natural host–parasite system including a fish host and a trematode parasite to study the effects of G × G interactions on infection virulence. We exposed rainbow trout (Oncorhynchus mykiss) either to single genotypes or to mixtures of two genotypes of the eye fluke Diplostomum pseudospathaceum and estimated parasite infectivity (linearly related to pathogenicity of infection, measured as coverage of eye cataracts) and relative cataract coverage (controlled for infectivity). We found that both traits were associated with complex G × G interactions, including both increases and decreases from single infection to co‐infection, depending on the genotype combination. In particular, combinations where both genotypes had low average infectivity and relative cataract coverage in single infections benefited from co‐infection, while the pattern was opposite for genotypes with higher performance. Together, our results show that infection outcomes vary considerably between single and co‐infections and with the genetic identity of the co‐infecting parasites. This can result in variation in parasite fitness and consequently impact evolutionary dynamics of host–parasite interactions.     

微生态治疗老年性阴道炎的临床观察

目的探讨采用乳杆菌活菌胶囊治疗老年性阴道炎的临床效果。方法将50例绝经妇女诊断为老年性阴道炎患者随机分成两组,研究组26例每晚清洁外阴后,阴道深部放置乳杆菌活菌胶囊500mg,连续10d。对照组24例每晚阴道内置人甲硝唑500mg,连续10d。两组均在停药后第3-7天复查比较其治愈率、总有效率。两组治愈者均于停药后90d复查,比较其复发率。结果研究组治愈率为73．07％,总有效率为96．14％。对照组治愈率为62．50％,总有效率为91．66％,两组差异无显著性（P〉0．05）。研究组复发率为0,对照组复发率为20．83％,两组差异有显著性（P〈0．05）。结论乳杆菌活菌阴道胶囊恢复阴道微生态平衡,增加阴道黏膜抵抗力。与抗生素药物比较复发率低,又无毒副作用,是治疗老年性阴道炎首进药物。     

Antioxidants and cataract

Abstract

The major causes for cataract formation are free radicals, and these free radicals are neutralized by the presence of endogenous antioxidants in the eye. Using xenobiotics, it has been confirmed that free radicals mediate the formation of cataract. Two cataract model-selenite model and the diabetic cataract model-have been developed to study the pathophysiology of cataract formation due to free radicals and the role of antioxidants during the process of cataractogenesis. This review focuses on natural compounds with antioxidant properties that could actually be applied as an interventional strategy on a large scale and are also relatively inexpensive. A brief overview of plants with antioxidant properties that in addition possess potential anti-cataract properties has been discussed. In addition to plants, three natural compounds (curcumin, vitamin C and vitamin E), on which a lot of data exist showing anti-cataract and antioxidant activities, have also been discussed. These antioxidants can be supplemented in the diet for a better defence against free radicals. Studies on vitamin C and vitamin E have proved that they are capable of preventing lipid peroxidation, thereby preventing the generation of free radicals, but their efficacy as anti-cataract agent is questionable. Unlike vitamins C and E, curcumin is well established as an anti-cataract agent, but the issue of curcumin bioavailability is yet to be addressed. Nanotechnology proves to be a promising area in increasing the curcumin bioavailability, but still a lot more research needs to be done before the use of curcumin as an effective anti-cataract agent for humans.     

分离自老年病房的鲍曼不动杆菌耐药机制初步研究*

目的:检测老年住院患者分离的鲍曼不动杆菌的主要耐药基因,并研究不同耐药基因型与耐药表型之间的对应关系。方法:用PCR方法检测分离自老年住院患者的不同标本来源的170例非重复鲍曼不动杆菌的耐药基因。检测的耐药基因包括D类碳青霉烯酶:bla_(OXA-51),bla_(OXA-23),bla_(OXA-24),bla_(OXA-58),B类金属碳青霉烯酶:bla_(VIM),bla_(IMP),bla_(SIM),bla_(GIM),bla_(DIM),bla_(NDM-1),以及A类超广谱β-内酰胺酶:blaKPC,共计11种。根据检测结果对菌株进行基因分型,并研究不同基因型与CRAB和CSAB这两种耐药表型之间的对应关系。结果:170株鲍曼不动杆菌的固有基因bla_(OXA-51)均为阳性,此外,主要检出基因为bla_(OXA-23),共124株。另外检测出blaKPC12株,bla OXA-58 6株,bla_(NDM-1)3株,bla_(SIM)2株,bla_(OXA-24)、bla_(VIM)和bla_(DIM)各1株,IMP和GIM未检出。根据检出耐药基因的不同组合,分为bla OXA-51+bla_(OXA-23)阳性为基础的A型(124株)及bla_(OXA-23)阴性为基础的B型(bla_(OXA-51),39株)、C型(bla_(OXA-51)+bla_(OXA-58),6株)、D型(bla_(OXA-51)+bla_(OXA-24),1株)共计四类基因型。从耐药表型来看,128株碳青霉烯耐药菌中有122株bla_(OXA-23)为阳性,在CRAB中占95.3%(122/128),42株碳青霉烯敏感株中,有40株bla_(OXA-23)为阴性,在CSAB中占95.2%(40/42)。结论:老年病房流行的耐碳青霉烯鲍曼不动杆菌的耐药基因型以bla_(OXA-23)阳性为主。其与鲍曼不动杆菌CRAB耐药表型、bla_(OXA-23)阴性与CSAB耐药表型之间有良好的对应关系。     

老年患者血培养病原菌分布及耐药性调查

目的了解老年患者血液感染的病原菌种类以及对抗菌药物的耐药状况。方法对2290份老年住院患者血培养标本中186例培养阳性检出菌及药敏结果进行统计学分析。结果在分离的172株细菌中,革兰阴性菌、革兰阳性菌及真菌分别占54.7%、34.9%和10.5%;革兰阴性菌中以大肠埃希菌占绝对优势(20.9%),假单胞菌其次(10.5%),革兰阳性菌中以凝固酶阴性葡萄球菌(CNS)占绝对优势(20.9%),金黄色葡萄球菌仅为2.9%。药敏试验表明老年菌血症的病原菌对常用抗菌药物的耐药严重,尤其是CNS及假单胞菌属细菌表现为较高的耐药率及多重耐药性。结论老年菌血症病原菌以机会致病菌感染比例较高,致病菌耐药问题严重,及时监测老年致病菌的变迁和耐药发展趋势以指导临床用药至关重要。     

盐酸多奈哌齐联合综合康复训练对老年痴呆患者认知功能、事件相关电位及血清BDNF、IGF-1的影响

目的:探讨盐酸多奈哌齐联合综合康复训练对老年痴呆患者认知功能、事件相关电位及血清脑源性神经营养因子(BDNF)、胰岛素样生长因子1(IGF-1)的影响。方法:选取2017年3月~2020年3月期间来我院接受治疗的120例老年痴呆患者,根据随机数字表法分为对照组(n=60,盐酸多奈哌齐治疗)和联合组(n=60,盐酸多奈哌齐联合综合康复训练治疗)。对比两组疗效,治疗前、治疗3个月后的认知功能评分、事件相关电位、血清BDNF和IGF-1水平变化,以及治疗期间不良反应发生情况。结果:联合组的临床总有效率为91.67%,高于对照组的70.00%(P<0.05)。两组治疗3个月后日常生活能力评估量表(ADL)、临床痴呆量表(CDR)评分降低,且联合组低于对照组(P<0.05);简易精神状态量表(MMSE)评分升高,且联合组高于对照组(P<0.05)。两组治疗3个月后N1潜伏期、P2潜伏期均缩短,P3波幅均升高,且联合组治疗3个月后P2潜伏期短于对照组,P3波幅高于对照组(P<0.05);两组治疗3个月后N1潜伏期比较未见统计学差异(P>0.05)。联合组治疗3个月后BDNF水平高于对照组(P<0.05);两组治疗3个月后IGF-1水平比较未见统计学差异(P>0.05)。对照组与联合组不良反应发生率对比无统计学差异(P>0.05)。结论:老年痴呆患者在盐酸多奈哌齐基础上联合综合康复训练,认知功能可得到明显改善,同时还可调节患者事件相关电位及血清BDNF、IGF-1水平。