BackgroundBigbag intraocular lens (IOL),due to its special conducive design to support vitreous and retina after cataract surgery in high myopia,and ensure the visualization of peripheral retinal,is closely concerned.But whether its concave design has the advantage in reducing aberrations on the basis of stability or not is worth to be studied.ObjectiveThis study was to evaluate the high order aberrations(HOA) of phacoemulsification and the Bigbag IOL implantation in patients with myopia and cataract.MethodsA retrospective case-observational study was designed.Total 39 eyes of 39 cases with cataract and high myopia were enrolled in this study.The patients were divided into Bigbag IOL group and AR40e IOL group.Phaco-chop technique and IOL implantation were performed in April to June,2010 in Tianjin Eye Hospital.Two months after surgery,the operated eyes were clinical examined and the aberrations were detected including Root-Mean-Square (RMS),coma,spherical and Trefoil.Postoperative RMS values of HOA components of the whole and interior optic with iTrace were compared.Results The operation was smoothly with the correct IOL position after two months.No complication was found.Under the 4.5 mm pupil diameters,the whole ocular HOA RMS values and coma were significantly different between Bigbag IOL group and AR40e IOL group( t =-3.296 、-3.322,P＜0.05 ),but no significant differences were seen in spherical and Trefoil aberrations ( t =- 1.256、- 0.573,P＞ 0.05 ).Regarding to the interior optic with iTrace aberration,only the coma showed the significant difference between Bigbag IOL group and A R40e IOL group( t =-2.004,P＜0.05 ),and there were no significant differences in RMS,spherical and Trefoil aberrations( t=-1.073、0.380、0.288,P＞0.05 ).ConclusionsThe Bigbag IOL,a design for high myopia,is safe and effective after implant for the high myopia with cataract.It offers more exact obligate degree and better visual quality.The chief total and interior HOA components difference between groups is coma aberration.

Background The test for visual acuity is the conventional standard for evaluating visual quality.However,there is little correlation between visual acuity and visual disability.Therefore,other clinical observations such as contrast sensitivity or straylight have been used in the clinical assessment of visual quality after cataract surgery.Objective This study was to examine the change in the amount of straylight in cataractous eyes and to evaluate the correlation of the types of cataract with visual quality.Methods A non-randomized case-controlled study was designed.Eighty eyes of 40 patients with age-related cataract were enrolled in this study,including 22 eyes with hard nucleus cataract,19 eyes with cortical cataract,23 eyes with mixed cataract and 16 eyes with posterior subcapsular cataract diagnosed based on the criteria from Lens Opacities Classification System Ⅲ(LOCS Ⅲ).The values for straylight and best corrected visual acuity (BCVA) were measured with the C-quant straylight meter and Snellen chart,respectively.The differences in the straylight values among the different types of cataract were analyzed,and the correlation of the straylight value with age or BCVA was assessed.Forty eyes of 40 age-matched normal people served as controls.Results Examination was completed in sixty-two eyes of 31 patients in the cataract group at a completion rate of 77.5％,and all the subjects in the control group finished the examination at a rate of 100％.The mean straylight value was (2.06±0.88) log in the cataract group and (1.96±0.42) log in the control group,showing a significant difference between them (t =3.251,P＜0.01).The respective mean C-quant measurements for hard nucleus cataract,cortical cataract,nuclear-cortical cataract and posterior subcapsular cataract patients were (1.96±0.42) log,(1.91 ±0.16) log,(2.05 ±0.19) log and (2.48 ±0.66) log,respectively,with a significant difference among these four groups (F =2.156,P =0.019).The highest straylight value was detected in the posterior subcapsular cataract group.The straylight value was enlarged with the increase of age with a regression equation of Y=0.0010X+ 1.025 in the hard nucleus cataract group (r =0.455,P ＜ 0.05).In addition,the negative linear correlation was found between the straylight value and BCVA in both the hard nucleus cataract group and cortical cataract group (r=-0.590,-0.697,P＜0.01).However,no correlation was found in the mixed cataract group and posterior subcapsular cataract group (r =-0.240,-0.235,P＞0.05).Conclusions The C-quant straylight meter can objectively reflect the visual function for hard nucleus and cortical cataractous eye.Posterior subcapsular cataract produces straylight and exerts a great influence on visual quality due to early glare sensation,so it should be benefit to perform surgery earlier.

Objective To explore the effect of Mycobacterium tuberculosis antigen (Mtb-Ag) on neutrophils apoptosis.Methods The fresh isolated neutrophils from healthy adults blood were cultured with Mtb-Ag for 24 h,with or without pretreatment of nuclear factor -?B (NF-?B) inhibitor N-tosyl-L-phenylanyl chloromethyl ketone (TPCK) for 30 minutes.Annexin V staining and Flow cytometry were used to measure cell apoptosis of neutrophils.NF-?B DNA binding was measured by gelelectrophorestic mobility shift assay (EMSA) in neutrophils after incubated with Mtb-Ag for 0,1,2,4,6,24 hours.Results Comparing to the spontaneous apoptosis (55%?6%) of neutrophils after culture in vitro for 24 h,treatment of Mtb-Ag (1.125 mg/ml) decreased the cell apoptosis of neutrophils (32%?3%).The NF-?B shift bands were detected at 1 h in neutrophils after stimulated by Mtb-Ag,and reached maximum peak at 2 hours,and then returned to basal levels within 24 h.Pretreatment of TPCK inhibited the anti-apoptosis role of Mtb-Ag in neutrophils.Conclusion Mtb-Ag prevents neutrophils apoptosis and its inhibitory role concerns NF-?B pathway.

Objective To explore the method to develop a gene vaccine of human cytomegalovirus (HCMV) phosphoprotein 65 (pp65) against its infection. Methods HCMV strain AD169 was propagated in WI-38 cell and viral DNA was extracted as a template for polymerase chain reaction (PCR) amplification of UL83 (pp65), the resulting of PCR was subcloned into pUC118HincII/BAP plasmid and DNA sequence analysis conformed the fidelity of the PCR. The vector pcDNA 5.0 was designed to correctly place CMV promoter sequence, pp65 sequence and secret signal sequence (mouse immunoglobulin kappachain for efficient secretion of recombination protein) into its genomic DNA. Exchanged primers of pp65 sequence, CMV promoter sequence and secret signal sequence to confirm the result by PCR screening. The vector pcDNA 5.0 was transfected into CHO cell, supernatants of transfected cells were extracted and purified. Recombination protein from supernatants was detected by gel electrophoresis and dot blot hybridization of Western- ECL system. Results Compared the sequence of pp65 gene with the standard sequence of pp65 from Medline,it was found that the concordant rate between them was 99.99%,only a nonsense mutation occurrences at 1 455 base.A pcDNA 5.0 Eukaryotic expression vector was established, which including CMV promoter sequence,secretion signal sequence and pp65 sequence. PCR screening and the pp65 protein expressed in CHO cell confirmed it. Extraction from supernatants in transfected CHO cells was recombination protein of pp65, which was detected by gel electrophoresis and dot blot hybridization of Western- ECL system and western blotting.Conclusion Subunit vaccine of HCMV is gained,which is a transfer eukaryotic expression vector pcDNA 5.0 constructed by CMV promoter sequence,secretion signal sequence and pp65 gene sequence.

OBJECTIVE:To establish an RP-HPLC method for content determination of s-omeprazole sodium and its related substances.METHODS:The separation of s-omeprazole sodium and the related substances was carried out on a Phenomenex Luna C18 column,the mobile phase consisted of methanol-0.033 mol?L-1 ammonium dihydrogen phosphate-triethylamine (58∶41.8∶0.2,adjusted to pH 7.0 by phosphate acid).The detection wavelength was 302 nm,the flow rate was 1.0 mL?min-1,the column temperature was 25 ℃,and the sample size was 20 ?L.RESULTS:The linear range of omeprazole sodium was 10～500 mg?L-1 (r=0.999 7).The average recovery rate was 100.27% (RSD=0.74%).The average content of the related substances in samples was 0.42%.CONCLUSION:This method is simple,accurate,specific and applicable for content determination of s-omeprazole sodium and its related substances.

OBJECTIVE:To establishment the method for the determination of content of vitamin A palmitate(VAP) and its related substances by HPLC. METHODS:The HPLC conditions were consisted of Phenomenex Luna C18 column with a mobile phase of a mixture of acetonitrile-isopropanol (90∶10) ,the detection wavelength of 328 nm,the column temperature of 30 ℃ and the flow rate of 1.0 mL?min-1. RESULTS:VAP was completely separated from impurities,the linearity range was 90～400 mg?L-1(r=0.999 2). The average recovery rate was 99.60% (RSD=1.32%). The average content of the related substances were lower than 2.53% . CONCLUSION: This accurate and reliable HPLC method is applicable for the quality control of VAP.

This study was aimed to examine the effect of TREK-1 silencing on the function of astrocytes. Three 21-nucleotide small interfering RNA (siRNA) duplexes (siT1, siT2, siT3) targeting TREK-1 were constructed. Cy3-labeled dsRNA oligmers were used to determine the transfection efficiency in cultured astrocytes. TREK-1-specific siRNA duplexes (siT1, siT2, siT3) at the optimal concentration were transfected into cultured astrocytes, and the most efficient siRNA was identified by the method of immunocytochemical staining and Western blotting. The proliferation of astrocytes tranfected with TREK-1-targeting siRNA under hypoxia condition was measured by fluorescence-activated cell sorting (FACS). The results showed that TREK-1 was expressed in cultured astrocytes. The dsRNA oligmers targeting TREK-1 could be transfected efficiently in cultured astrocytes and down-regulate the expression of TREK-1 in astrocytes. Moreover, the down-regulation of TREK-1 in astrocytes contributed to the proliferation of astrocytes under hypoxia condition as determined by cell cycle analysis. It was concluded that siRNA is a powerful technique that can be used to knockdown the expression of TREK-1 in astrocytes, which helps further investigate the function of TREK-1 channel in astrocytes under physicological and pathological condition.

Objective To investigate BAALC(brain and acute leukemia cytoplasmic)gene expression in patients with de novo acute myeloid leukemia(AML)and its clinical significance. Methods BAALC expression was determined by real-time quantitative polymerase chain reaction(RQ-PCR) in 63 de novo AML patients.The association between BAALC expression and therapeutic effect was analyzed.Results The correlation coefficiencies were over 0.99 for standard curves of RQ-PCR method. BAALC expression was detected in 49(78%)AML patients.The peripheral WBC counts,hemoglobin, platelet counts and the bone mahow blast cell percentage at onset in 31 AML patients with high BAALC expression were(26.3?18.1)?10~9/L,(78.3?21.8)g/L,(76.9?64.5)?10~9/L and(61.2?22.3)% and those of 32 AML patients with low BAALC expression were(30.2?21.7)?10~9/L,(81.6?30.9)g/L, (73.9?57.2)?10~9/L,(54.3?16.3)%,respectively.No statistic differences were found between these two groups.The AML patients with normal chromosome karyotypes are more likely to have a high BAALC expression(68%)compared with those with abnormal chromosome karyotypes(23%,?~2=12.093,P= 0.001).AML patients with normal cytogenetics and high BAALC expression shows significant lower CR rate (65%)compared with those with low BAALC expression(84%,?~2=6.573,P=0.013). Conclusion High BAALC expression may define an important risk factor in AML with normal cytogenetics and predicts an adverse prognosis.

Objective To study the changes of phosphatidylcholine-specific phospholipase D (PC-PLD) enzyme activity of glomerular mesangial cells (GMCs) in high glucose and the effect of PLD on cytoskeleton. Methods Rats GMCs were cultured in high glucose(30 mmol/L) for 48 hours. PLD enzyme activity was measured by enzyme coupled colorimetry. PKC activity was measured by substrate phosphorylation, and fluorescence intensity of F-actin was observed by FITC-labeled antibody and laser scan cofocal microscopy (LSCM) . Results In high glucose, PLD and PKC activities were obviously elevated, but fluorescence intensity of F-actin was decreased and its cytoskeletal pattern was disassembly. After treatment with inhibtor of PLD, PKC activity was significantly decreased, and intensity and organization of F-actin were recovered. Conclusion Elevation of PLD activity induced by high glucose may influence the cytoskeleton and contraction of CMC through PKC pathway.

OBJECTIVE To study the pharmacokinetics of heat shock protein 65-mucin 1 (HSP65-MUC1) recombinant fusion protein vaccine in Macaca mulatta monkeys and tumor-bearing mice. METHODS HSP65-MUC1 was labeled by radioactive isotope 125I. M. mulatta monkeys were randomly divided into sc and iv administration groups. Simultaneously, sc administration group was designed as a multiple dose group in which M. mulatta monkeys were sc given [ 125I] HSP65-MUC1 40 μg·g-1, once every 2 weeks for a total of 3 times. Size exclusion chromatography ( SEC) was used to determine concentrations of HSP65-MUC1 in serum samples. The tumor-bearing mice were randomly divided into 0.5, 1.5, 4, 8 and 24 h groups. Mice were sc given [125I] HSP65-MUC1 550 μg·kg-1, tissues were collected and tissue distribution of [125I] HSP65-MUC1 in tumor-bearing mice was studied using trichloroacetic acid (TCA) precipitation method. RESULTS The absolute bioavailability of [125I]HSP65-MUC1 was 38.33% after M. mulatta monkeys were sc given [125I]HSP65-MUC1. In multiple dose group, concentrations of [125I]HSP65-MUC1 after the third dose administration was compared to that of the first dose administration. The accumulation factor (AUC3/AUC1) was 1.17 ±0.25. Distribution of [ 125I]HSP65-MUC1 was significantly different compared with general polypeptide and protein drugs after sc in tumor-bearing mice. The concentration in lymph nodes was the highest. The concentration in other immune tissues, such as thymus and spleen, were not relatively high, but their declined tendency was slow after reaching the peak concentration (cmax ). However, the concentrations in the serum and some other tissues with a large blood volume, such as the heart, liver, and lung, were relatively low and declined quickly after reaching cmax. Its level in the tumor was not very high. [125 I] HSP65-MUC1 was excreted mainly by the kidneys. CONCLUSION The bioavailability of [125I]HSP65-MUC1 is 38.33% after sc administration in M. mulatta. After multiple-dose administration, the vaccine does not accumulate in the body, whose concentration is the highest in lymph nodes after [1251] HSP65-MUC1 was sc given in tumor-bearing mice, but is not very high in tumor. Besides, the vaccine declined tendency is slow after reaching cmax in immune tissues such as thymus and spleen compared with other tissues with a large blood volume.