Replacement therapy of stem cells transplantation represents a potential treatment for neural retinal diseases.Despite the encouraging results in laboratory,the clinical application of cells replacement therapy is still difficult because the limitation of seed cells,immunologic rejection,oncogenicity and ethical problems,etc.Recent breakthrough in somatic reprogramming provides a promising solution overcoming these obstacles.Further researches on virus-free reprogramming will make the clinical application of stem cell replacement therapy possible.

Glaucoma is a common blinding eye disease mainly characterized by optic nerve damage.Lowing intraocular pressure is still the main managing method of glaucoma up to now.However,although the intraocular pressure is reasonably controlled,optic nerve damage is difficult to recovery and sustainable development in some patients.Therefore,the pathogenesis and prevention and treatment of glaucoma have always been the hot spot of the research of glaucoma in recent years.Even though the big progresses have been made in the experimental research of optical nerve injury and repair,little good evidence has been seen in the clinical management of glaucoma neuroprotection.In recent years,genomics study,stem cells study,molecular biological study,electronic technology application in medical research,especially the rise of big data era laid a good basis for the neuroprotection of glaucoma.Ophthalmologists should focus on new insights into the potential and beyond impact factors in the precise individual therapy of glaucoma neuroprotection.

Glaucoma ; diagnosis ; therapy ; Humans ; Research

The following progresses have been recently made on glaucoma in Zhongshan Ophthalmic Center,covering clinical and basic researches on molecular genetics and epidemiology,stem eells,pathogenesis of calcium and adenosine,neuroprotection,etc. New insight has been shown into about 30 papers published in international peer-review(SCI)journals.

The paper briefly reports the preliminary research results,focusing on the pathogenesis,mechanisms and interference strategies of glaucoma and myopia.(1)The myocilin gene has been found a mutation in Pro370Leu in GZ.1 and PN.1 glaucoma pedigrees,and the theory of protein misfolding and mitochondria dysfunction in glaucoma was then firefly elucidated.We further explored the modulation of wound healing by siRNA,neuro-protection by targeting drugs with drug deliver system and stem cells/iPS in glaucoma therapy,which offer a revolutionary strategy for glaucoma:differential treatment for specific stages.(2)Natural myopia model was successfully established in rhesus monkeys and retinal mechanisms and central nervous system were both found to play an important role during myopia development,and a variety of new optical interventions were therefore employed on human beings by series of randomized,controlled,double-masked prospective clinical trials.All these preliminary results offer a new insight into the new model for the diagnosis and treatment of glaucoma and myopia.

OBJECTIVE:To study the effects of grape seed proanthocyanidin(GSP)on the transmembrane transport of sodium glycocholate (GA) and sodium taurocholate (TA) in colon glandular cell Caco-2. METHODS:Caco-2 model was used,and RP-HPLC was conducted to determine the contents of GA and TA in cell culture medium. The test was divided into GSP group, GA group,TA group,GSP+GA group and GSP+TA group,the transport volumes of transporting GA and TA from Transwell apical (AP)side to basolateral(BL)side by Caco-2 cell at 0,2,4,8 h were detected,respectively. RESULTS:The linear ranges of GA and TA were 0.05-1.2 mmol/L(R2=0.9999). With the time passing,transport volumes of GA and TA in BL site in GA group and TA group were sharply increased;while the transport volumes were obviously decreased after adding GSP,with statistical signifi-cance(P<0.01). CONCLUSIONS:GSP has inhibitory effect on the transmembrane transport of GA and TA in Caco-2 cell.

strains that could produce raw starch-digesting glucoamylase were isolated from soil and mildewed rice.The highest raw starch-digesting glucoamylase activity strain named OR-1 was identified as Rhizopus.sp.The raw starch-digesting glucoamylase activity of the strain is 90U/mL.Through UV and NTG mutagenesis,the raw starch-digesting glucoamylase activity raised to 200U/mL and 325U/mL respectively.The RDA were 70% and 65% respectively.

ObjectiveTo detect the expression on p53 gene in retinal ganglion cells(RGCs) of rabbit eyes with different degrees of intraocular hypertension.MethodsIntraocular pressure(IOP)was elevated by intracameral injection of 0.10～0.15 ml of 2% methylcellulose.Animals were grouped up by different values of IOP.The rabbit eyes were enucleated,fixed,paraffin-embedded and cut into serial 6- μm thin sections.For histological evaluation,the slides were stained with DAB chromogen(conducting an immuohistochemical experiment),and were examined by light microscopic and automatic image analysis.ResultsThe number of RGCs with a positive reaction for p53 monoclonal antibody was statistically higher in the experimental eyes than in their control fellow eyes(P<0.01),and apoptosis was mainly found in the early stage of research.ConclusionOur research presents a new alternative view of apoptosis in experimental glaucoma(According to former reports,apoptosis was mainly found in the advanced stage of the patients with POAG)

BACKGROUND:Previous studies have suggested that achaete-scute homology 1 (ASCL1) plays a key role in the neuronal commitment. Therefore, somatic cels may directly differentiate into neurons by gene transfection ofASCL1, which wil provide new therapeutic strategies for optic nerve regeneration. OBJECTIVE:To construct a recombinant adenovirus vector expressing ratASCL1 gene for further research ofASCL1 gene function. METHODS:The ratASCL1 gene and advenovirus shuttle plasmid (pYr-adshuttle-4) which contained enhanced green fluorescent protein (EGFP) reporter gene were cleaved by restriction endonucleaseXhoI andEcoR I. The target gene fragments were connected together to generate a recombinant plasmid pYr-ads-4-rat-ASCL1 and then transfected into E.coliDH5α. The plasmid was confirmed to be constructed as expectation by enzyme digestion and sequence reaction. The plasmid pYr-ads-4-rat-ASCL1 and pAd/PL-DEST were reconstructed by homologous recombination processes to obtain rat ASCL1 recombinant adenovirus vector. The plasmid pYrAd-rASCL1 was linearized byPac I and subsequently transfected into HEK293 cels for packaging and amplification. RatASCL1 gene in the recombinant adenoviruses were identified by PCR. Virus titer was determined by tissue culture infectious dose 50. Infection efficiency was monitored by EGFP expression. RESULTS AND CONCLUSION:Restriction endonuclease digestion and DNA sequencing showed that the recombinant adenovirus vector pAd-rat-ASCL1 was constructed correctly. The positive amplification bands of 862 bp could be seen in PCR analysis. The virus titer reached 2×1010 pfu/mL. Infection efficiency of recombinant adenovirus in HEK293 cels was more than 80%. The results indicate that the recombinant the adenovirus vector containingASCL1 with high titer and infection efficiency has been successfuly constructed, which can be helpful for further research of the function and clinical application ofASCL1 gene for optic nerve regeneration.