1.The regulation of estradiol on growth dynamics of human LECs affected by increasing telomerase activity
Jie, WANG ; Gangjin, KANG ; Xuefeng, YUAN ; Manhua, XU ; Yan, JIANG ; Bo, LUO
Chinese Journal of Experimental Ophthalmology 2016;34(3):219-225
Background Human LECs can express telomerase activity,which participates in the formation of cataract.It is reported that estrogen can increase the expression of telomerase activity in human endometrial cancer and breast cancer cells and play an important role in promoting proliferation and anti-apoptosis,but whether estrogen exerts its role on human LECs is still unclear.Objective This study aimed to investigate whether β-estradiol (β-E2) can increase the telomerase activity of human LECs and the influence of β-E2 on proliferation and apoptosis of human LECs.Method Human LECs line was cultured and passaged in vitro,and 1×10-6 mol/L β-E2 was added in the medium for 0,6,12,24 and 48 hours,and reverse transcription PCR was used to determine the optimal time of the expression of human telomerase reverse transcriptase (hTERT) mRNA in the cells.Cultured cells were divided into five groups.The cells in the blank contol group were cultured in the routin medium.Ethanol of 0.1% was added in the solvent control group,and 1 × 10-8,1 × 10-7 or 1 × 10-6 mol/L β-E2 was added in the medium in different contents of β-E2 groups,respectively.The relative expression level of hTERT mRNA in different groups was detected by reverse transcription PCR.Telomere repeat amplification protocol (TRAP)-ELISA was employed to determine the telomerase activity.The proliferative value of the cells was assayed by cell counting kit-8,and the apoptosis rate of the cells was examined by Hoechst33258 staining.Results The optimal time of β-E2 to rise the expression of hTERT mRNA (absorbance) was at 24 hours under the 1×10-6 mol/L.The relative expression levels of hTERT mRNA in the cells were 0.477±0.015,0.712±0.013 and 0.914±0.031 in the 1 ×10-8,1 ×10-7 and 1 ×10-6 mol/L β-E2 group,which were signifincatly higher than 0.428±0.010 in the blank control group and 0.426±0.010 in the solvent control group (all at P<0.05).The telomerase activity values (absorbance) were 0.711 ±0.015,0.941±0.010 and 1.249±0.047 in the 1×10-8,1×10-7and 1×10-6 mol/L β-E2 group,which were higher than 0.535±0.013 in the blank control group and 0.543 ±0.013 in the solvent control group (all at P =0.000).The proliferantive values of the cells (absorbance) were significantly raised in the 1 × 10-8 mol/L β-E2 group compared with l × 10-7 and 1 × 10-6 mol/L β-E2 group (both at P =0.000),and no significnant difference was found in the proliferetive values between the blank control group and the solvent control group (P =0.718,0.856).The apoptosis rates of the cells in the 1 × 10-8,1 × 10-7 and 1 × 10-6 mol/L β-E2 group were lower than those in the the blank control group and the solvent control group (all at P=0.000),and there was no significant difference between the blank control group and the solvent control group (P =0.777).No obvious correlation was found between the HLECs preliferative values and hTERT mRNA expression levels or telomerase activity values (r=-0.299,P=0.278;r=-0.157,P=0.576).However,significantly negative correlations were seen between apoptosis rates and hTERT mRNA expression levels or telomerase activity values (r =-0.975,P=0.000;r=-0.981,P=0.000).Conclusions β-E2can increase the activity of telomerase in human LECs,and high dose of β-E2can inhibit apoptosis,but it dose not promote proliferation.
2.Cross-cultural adaptation, preliminary validity and reliability and modification of the Chinese Version of the KING′s Parkinson′s Disease Pain Scale
Hanying GU ; Chengjie MAO ; Fen WANG ; Xiaoyu CHENG ; Ming LIU ; Manhua LIU ; Yajun LUO ; Yiming WANG ; Chunfeng LIU
Chinese Journal of Neurology 2022;55(9):968-978
Objective:To perform cross-cultural adaption of the KING′s Parkinson′s Disease Pain Scale (KPPS), explore its reliability and validity in Chinese Parkinson′s disease (PD) patients, and to create the new version of the pain scale which adapts to the Chinese PD patients.Methods:This study enrolled 225 patients, including 121 men and 104 women who were selected from the Outpatient Center of Movement Disorders Clinic of the Second Affiliated Hospital of Soochow University from July 2018 to July 2020. All patients completed the evaluation of the Chinese Version of KPPS (KPPS-CV). According to the preliminary evaluation results, the expert group modified KPPS-CV after discussion, and developed a Modified KPPS-CV (MKPPS-CV). These patients then completed the MKPPS-CV evaluation during the 3-month follow-up. Cross-cultural adaptation was performed according to published international guidelines that include translation, back-translation, expert review, and pretesting. The following psychometric properties were evaluated: basic item analysis; floor and ceiling effects; construct validity; content validity; criterion validity (Spearman′s rho between the KPPS-CV and Numeric Rating Scale); internal consistency reliability (Cronbach′s alpha); test-retest reliability (intra-class correlation coefficient, ICC).Results:In item analysis, 50% of the items had poor discrimination (critical ratio<3.0), and floor effect was found in all domains (proportion of 0 point>15%). The items were reclassified after exploratory factor analysis. The content validity of item 3, item 10 and item 11 was low (item-level content validity index<0.78). Criterion validity showed the highest correlations (Spearman′s rho>0.88) between the KPPS-CV and Numeric Rating Scale. While overall scale reliability was minimally acceptable at 0.46, which showed a poor reliability of this scale. Test-retest reliability was excellent for each item (Spearman's rho>0.85). The Cronbach′s alpha of MKPPS-CV (0.76) was higher than that of KPPS-CV (0.46). It showed a great improvement after the modifying.Conclusions:When using scales that are not developed for local populations, differences in culture and clinical practices should be taken into account. MKPPS-CV is an acceptable, valid measure to evaluate pain in Chinese PD patients, which is more suitable for Chinese people.
3.Observation on the effect of modified buffy coat centrifugal canning method on improving the quality of platelet concentrates
Manhua LUO ; Meiyan KANG ; Jin LI ; Xianbin WU
Chinese Journal of Blood Transfusion 2023;36(2):193-195
【Objective】 To evaluate the quality of platelet concentrates (PCs) prepared by traditional and modified buffy coat centrifugal canning methods, and to explore the effect of modified buffy coat centrifugal canning method on improving the quality of PCs. 【Methods】 The buffy coat centrifugal canning methods was divided into traditional group and modified group. In the traditional group, the buffy coat component bag and empty bag were directly combined layer by layer and vertically canned for light centrifugation. In the modified group, a foam spacer was added between the buffy coat component bag and the empty bag for light centrifugation layer by layer. The effects of the two groups of centrifugal canning methods on the preparation quality of PCs were observed. 【Results】 The platelet content in PCs prepared by the modified buffy coat centrifugal canning method was significantly higher than that in the traditional group, and the mixed amount of red blood cells was lower than that in the traditional group (P<0.05). Although there was no statistically significant difference in the qualified rate of platelet content between the two groups (P>0.05), the qualified rates of platelet content and mixed red blood cell in the modified group were higher than those in the traditional group (P<0.05). 【Conclusion】 The modified centrifugal canning method with buffy coat can improve the quality of PCs, which is convenient and cheap, and is worth popularizing.