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.

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.

【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.

Objective:Rejection remains the most important factor limiting the survival of transplanted kidneys.Although a pathological biopsy of the transplanted kidney is the gold standard for diagnosing rejection,its limitations prevent it from being used as a routine monitoring method.Recently,peripheral blood lymphocyte subpopulation testing has become an important means of assessing the body's immune system,however,its application value and strategy in the field of kidney transplantation need further exploration.Additionally,the development and utilization of routine test parameters are also important methods for exploring diagnostic strategies and predictive models for kidney transplant diseases.This study aims to explore the correlation between peripheral blood lymphocyte subpopulations and T cell-mediated rejection(TCMR)and antibody-mediated rejection(ABMR),as well as their diagnostic value,in conjunction with routine blood tests. Methods:A total of 154 kidney transplant recipients,who met the inclusion and exclusion criteria and were treated at the Second Xiangya Hospital of Central South University from January to December,2021,were selected as the study subjects.They were assigned into a stable group,a TCMR group,and an ABMR group,based on the occurrence and type of rejection.The basic and clinical data of these recipients were retrospectively analyzed and compared among the 3 groups.The transplant kidney function,routine blood tests,and peripheral blood lymphocyte subpopulation data of the TCMR group and the ABMR group before rejection treatment were compared with those of the stable group. Results:The stable,TCMR group,and ABMR group showed no statistically significant differences in immunosuppressive maintenance regimens or sources of transplanted kidneys(all P>0.05).However,the post-transplant duration was significantly longer in the ABMR group compared with the stable group(P<0.001)and the TCMR group(P<0.05).Regarding kidney function,serum creatinine levels in the ABMR group were higher than in the stable group and the TCMR group(both P<0.01),with the TCMR group also showing higher levels than the stable group(P<0.01).Both TCMR and ABMR groups had significantly higher blood urea nitrogen levels than the stable group(P<0.01),with no statistically significant difference between TCMR and ABMR groups(P>0.05).The estimated glomerular filtration rate(eGFR)was lower in both TCMR and ABMR groups compared with the stable group(both P<0.01).In routine blood tests,the ABMR group had lower hemoglobin,red blood cell count,and platelet count than the stable group(all P<0.05).The TCMR group had higher neutrophil percentage(P<0.05)and count(P<0.05)than the stable group,and the ABMR group had a higher neutrophil percentage than the stable group(P<0.05).The eosinophil percentage and count in the TCMR group were lower than in the stable and ABMR groups(all P<0.05).Both TCMR and ABMR groups had lower basophil percentage and count,as well as lower lymphocyte percentage and count,compared with the stable group(all P<0.05).There were no significant differences in monocyte percentage and count among the 3 groups(all P>0.05).In lymphocyte subpopulations,the TCMR and ABMR groups had lower counts of CD45+cells and T cells compared with the stable group(all P<0.05).The TCMR group also had lower counts of CD4+T cells,NK cells,and B cells than the stable group(all P<0.05).There were no significant differences in the T cell percentage,CD4+T cell percentage,CD8+T cell percentage and their counts,CD4+/CD8+T cell ratio,NK cell percentage,and B cell percentage among the stable,TCMR,and ABMR groups(all P>0.05). Conclusion:The occurrence of rejection leads to impaired transplant kidney function,accompanied by characteristic changes in some parameters of routine blood tests and peripheral blood lymphocyte subpopulations in kidney transplant recipients.The different characteristics of changes in some parameters of routine blood tests and peripheral blood lymphocyte subpopulations during TCMR and ABMR may help predict and diagnose rejection and differentiate between TCMR and ABMR.