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Background Blepharitis caused by Demodex infestation is very common in clinical practice.There are various methods mentioned in the study of Demodex infestation in China,but a unified introduction and evaluation of the operating procedures is lacked.A quick and accurate clinical diagnostic method for Demodex infestation needs to be further studied.Objective This study aimed to establish operation procedures for the clinical examination of eyelid Demodex infestation,which were applied to evaluate the conditions of eyelid Demodex infestation in ocular patients with discomfort.Methods One thousand and fifty-two patients with eye dryness,eye itchiness or other symptoms were selected for slit lamp examination and photographing of the eyelid margin area.Three eyelashes with associated scurf from each superior eyelid were plucked out for examination of Demodex under the microscope.Positive findings included observation of Demodex mites or eggs.Their amounts were recorded individually for all eyelash samples.Results A procedure for observing,recording and reporting eyelid Demodex infestations in patients was successfully established.By using this procedure,1 052 patients were investigated for the examination of Demodex infestations.Demodex mites or eggs were found in 582 cases (55.3％).The positive rate of Demodex infestation increased with age,and the population over 60 years group had the highest positive rate,showing a significant difference among the different age groups (x2=10.547,P=0.001).There was no significant difference in positive rate between male patients and female patients (P =0.352).The test turnaround time (TAT) for one examination was (11.4±5.2) seconds.Conclusions The operational procedure for examining the palpebral margin Demodex infestation by the slit-lamp,optical microscope,photographing and laboratory reports is established.It is simple and quick in the appliation for the clinical diagnosis of eyelid Demodex infestation.
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Objective To investigate the effect of N-methyl-N-nitrosourea (MNU) on expressions of interphotoreceptor retinoid-binding protein (IRBP) and recoverin in rat retinal.Methods Thirty female Sprague-Dawley (SD) rats were randomly divided into five groups,including normal control group,and MNU treatment one day,three days,seven days and 10 days groups,each group had six rats.Rats in MNU-treatment groups received a single intraperitoneal injection of 40 mg /kg MNU,while rats in the normal control group received intraperitoneal injection of saline 5 ml/kg.Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence were used to detect the retinal expression of IRBP and recoverin.Results RT-PCR results indicated that retinal IRBP levels decreased after MNU injection compared with normal control (P＜0.01),while recoverin levels increased (one day:P＞0.05; three days:P＜0.05; seven days:P＜0.5; 10 days:P＜0.05).Immunofluorescence assays demonstrated that normally IRBP protein is mainly in the inner and outer segments of photoreceptors.After MNU treatment,IRBP protein was detected in all retinal layers but much faint.Recoverin was expressed in the inner nuclear layer,inner plexiform layer and ganglion cell layer,and its expression was increased after MNU injection.Conclusion MNU suppresses IRBP expression and promotes recoverin expression in rat retina.
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It is kno wn that 8-Br-cAMP is one of selective bi nding site analogues for cAMP RIIα to af fect cell growth through regulation of g ene expression.The p16,p21wafl,p53 a nd Rb are antioncogenes which affect cel l growth through control of cell cycle.T he aim of this study is to investigate t he 8-Br-cAMP effect on the expression of antioncogenes in human HXO-Rb44 cells. Methods Cultured HXO-Rb44 cells in RPMI -1 640 medium were divided into two aliquot s.8-Br-cAMP (2×10-5mol/L) was added i nto one aliquot for 24h as the experime ntal group(EG),the another aliquot witho ut 8-Br-cAMP as the control group(CG).Af ter 24h,the cell suspension was dropped onto the nitrocellulose membrane.The mR NA of p16,p21wafl,wild type(w)p53,mut ant type(m) p53 and Rb were used respec tively with biotin-labeled cDNA probes b y intact cell RNA dot blot.The immunorea ctivity(IR) of P16,P21wafl,PRb,PCN A,cdk2 and cdk4 were detected respecti vely with specific monoclonal antibodies on dot blot.ResultsThe mRNA dot blot s ignals of mp53 and protein dot blot of cdk2-IR,cdk4-IR and PCNA-IR in EG were weaker than those in CG(P＜0.05～0.01). W hile,the mRNA signals of p16,p21wafl,wp53 and Rb in EG were stronger than tho se in CG(P＜0.05～0.01).The intensity of ea ch protein dot blot was consistent with that of their RNA dot blot (except for w P53-IR and mP53-IR not to be done).Conc lusions(1)8-Br-cAMP could up-regul ate expression of antioncogenes includin g p16,p21wafl,wp53,Rb,and protein exp ression of P16,P21wafl and PRb.(2) 8-Br-cAMP could down-regulate mp53 gene expression and protein expression of cd k2,cdk4 and PCNA.The results suggest t hat 8-Br-cAMP could inhibit human HXO-Rb 44 cell growth through interfering rela ted gene expression of cell cycle.
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Objective] To find a rapid method for diagnosing Acanthamoeba keratitis and identifing Acanthamoeba . [Methods] 10% potassium hydroxide(KOH) wet mount preparations, Acanthamoeba culture, inverted phase contrast microscopy,and pathological examination using H.E. staining and PAS staining. [Results]Using corneal scrapings and corneal materials obtained from surgery,7 cases and 5 cases of Acanthamoeba keratitis were diagnosed by 10% KOH wet mount preparations. 6 strains of Acanthamoeba were isolated in corneal materials of 6 cases by protozoa culture method. The cysts, trophozoites and pseudopods on the trophozoites of Acanthamoeba were directly observed under the inverted phase contrast microscope. The cysts and trophozoites of Acanthamoeba were seen by H.E. staining and PAS staining with 20 h. [Conclusion] Acanthamoeba keratitis could be rapidly diagnosed by 10% KOH wet mount preparations and inverted phase contrast microscopy. Acanthamoeba organisms could be directly observed and identified under inverted phase contrast microscope.