1.The proliferative inhibition and apoptosis promotion of Smac on human lens epithelial cells
Deqian, KONG ; Guangying, ZHENG ; Li, LI ; Yue, LIU ; Fangfei, CAI
Chinese Journal of Experimental Ophthalmology 2017;35(3):233-238
Background Posterior capsular opacification (PCO) is a primary complication after extracapsular cataract extraction.The mechanism of PCO is associated with proliferation,migration and epithelialmesenchymal transition (EMT) of human lens epithelial cells (LECs).To explore the target treatment of PCO is very important.Objective This study was to investigate the biological effects of second mitochondria-derived activator of caspases (Smac) on the proliferation and apoptosis of LECs.Methods Human LECs line (HLE-B3) and Smac-overexpressed LECs line were cultured,and the cells were transfected using small interfering RNA (siRNA)-Smac3 plasmid with green fluorescent protein (GFP) for 24 hours.Different concentration of transforming growth factor-β2 (TGF-β2) (5,10,20 and 50 μg/ml) or 200 μmol/L H2O2 were added respectively into the culture medium to establish PCO model and oxidative stress model.Cell counting kit-8 (CCK-8) assay was used to compare the cell proliferative activity among PBS group,TGF-β2 group and Smac-hyperexpression +TGF-β2 group.Flow cytometry was used to evaluate the apoptotic rate of the PBS group,H2 O2 group and siRNA-Smac+H2 O2 group.The expressions of Smac,caspase-3 and proliferating cell nuclear antigen (PCNA) mRNA and their proteins in the cells were detected by real-time quantitative PCR (RT-PCR) and Western blot.Results The GFP+ cells were≥ 80% 12 hours after siRNA-Smac3 transfection,with the optimal plasmid of siRNA-Smac3.GFP+ cell rate was (72.32 ± 2.31)% in the siRNA-Smac3 transfection group,which was significantly higher than that in the blank plasmid group ([4.91 ±0.24] %) (t=116.342,P<0.001).The relevant expression levels of Smac was 35.21 ±4.11 in the Smachyperexpression group,and that in the blank plasmid group was 15.24±2.48,with a significant difference between them (t =215.47,P<0.05).The cell viability of 20 ng/ml TGF-β2 affected PBS group,TGF-β2 group and Smachyperepression+TGF-β2 group was (98.4 ± 1.7) %,(98.9 ± 0.1) % and (64.2 ± 3.1) %,and the cell viability of Smac-hyperepression+TGF-β2 group was significantly lower in the Smac-hyperepression+TGF-β2 group than that in the TGF-β2 group (P<0.05).The apoptotic rate in the PBS group,H2 O2 group and siRNA-Smac+H2 O2 group were (2.9 ± 1.2) %,(45.1 ±4.5) % and (27.5 ± 1.8) %,and the apoptotic rate was evidently lower in the siRNA-Smac +H2O2 group than that in the H2O2 group (P<0.05).RT-PCR results showed that the expression levels of caspase-3 mRNA in PBS group,H2 O2 group and siRNA-Smac + H2 O2 group were 0.321 ± 0.103,0.715 ± 0.112 and 0.479 ±0.209,respectively.Compared with the H2 O2 group,the relative expression level of caspase-3 mRNA in siRNA-Smac+ H2O2 group was significantly decreased,the difference was statistically significant (P< 0.05).The PCNA mRNA expression levels in PBS group,TGF-β2 group and Smac-hyperepression+TGF-β2 group were 0.299±0.013,0.645± 0.102 and 0.490±0.209,respectively.Western blot results showed that the relative expression of caspase-3 protein in siRNA-Smac+H2O2 group and H2O2 group was 0.712±0.012 and 0.973±0.051,with significant difference between the two groups (t =132.52,P<0.05).The relative expression of PCNA protein in Smac-hyperepression+TGF-β2 group was 0.782±0.212,which was lower than 1.126±0.251 in the TGF-β2 group (P<0.05).Conclusions Smac may prevent and treat PCO by inhibiting the proliferation and promoting apoptosis of human LECs.
2.Effect of fluorescence method on cardiopulmonary function, immune function and safety during pulmonary segmental resection
Liang SHAN ; Guang YANG ; Deqian KONG
Chinese Journal of Postgraduates of Medicine 2023;46(6):548-552
Objective:To explore the effects of fluorescence method and improved expansion and collapse method on cardiopulmonary function, immune function and safety during segmental pneumonectomy.Methods:One hundred and fifty-six patients with lung segmental resection operation in Yucheng People′s Hospital from March 2018 to August 2020 were enrolled. They were divided into two groups according to the random number table method, each group with 78 patients. The patients in the observation group were treated with fluorescence method and the patients in the control group were treated with modified expansion collapse method. The operation indexes, immune function and cardiopulmonary function of the two groups were compared, and the incidence of postoperative complications were counted.Results:The intraoperative time and the appearance time of intersegmental plane in the observation group were shorter than those in the control group: (105.33 ± 10.62) min vs. (139.46 ± 12.58) min, (15.46 ± 1.13) s vs. (724.56 ± 65.65) s, there were statistical differences ( P<0.05). After operative for 1, 7 d, the levels of 6 min walking distance, peak expiratory flow rate, percentage of forced vital capacity and percentage forced expiratory volume in 1 second in the observation group were higher than those in the control group: after operative for 1 d: (475.36 ± 10.29) m vs. (412.11 ± 13.33) m, (284.52 ± 10.33) L/min vs. (251.13 ± 12.89) L/min, (85.65 ± 3.21)% vs. (81.13 ± 1.43)%, (83.25 ± 2.55)% vs. (74.49 ± 2.53)%; after operative for 7 d: (510.23 ± 16.66) m vs. (488.33 ± 15.42) m, (302.13 ± 15.58) L/min vs. (285.12 ± 10.22) L/min, (93.46 ± 5.79)% vs. (88.44 ± 5.44)%, (92.25 ± 2.32)% vs. (85.54 ± 2.13)%, there were statistical differences ( P<0.05). After operative for 1, 7 d, the levels of CD 4+/CD 8+, CD 8+, CD 4+ in the observation group were higher than those in the control group: after operative for 1 d: 0.85 ± 0.10 vs. 0.52 ± 0.04, 0.305 ± 0.025 vs. 0.285 ± 0.012, 0.325 ± 0.021 vs. 0.304 ± 0.025; after operative for 7 d: 1.13 ± 0.10 vs. 1.02 ± 0.07, 0.324 ± 0.029 vs. 0.306 ± 0.023, 0.359 ± 0.024 vs. 0.332 ± 0.025, there were statistical differences ( P<0.05). The rate of postoperative complications in the observation group was lower than that in the control group: 2.56%(2/78) vs. 11.54%(9/78), there was statistical difference ( χ2 = 4.79; P = 0.029). Conclusions:Both the fluorescence method and the modified inflation-collapse method have certain therapeutic effects, but the combination of fluorescence method in segmentectomy can provide reliable technical support and reduce the impact on cardiopulmonary function.
3.Long-term clinical effect of aspheric multifocal intraocular lens implantation for developmental cataracts in childhood
Li LI ; Guangying ZHENG ; Yating ZHAO ; Yun LI ; Deqian KONG
Chinese Journal of Experimental Ophthalmology 2018;36(3):209-214
Objective To observe the long-term visual quality and clinical effect of aspheric diffractive multifocal intraocular lens (IOL) implantation for congenital and developmental cataracts in childhood.Methods A retrospective cohort study on multifocal IOL implantation for congenital and developmental cataracts in childhood (aged 8 to 14 years) was performed in the First Affiliated Hospital of Zhengzhou University from August 2013 to January 2015.The clinical data of 67 eyes from 46 congenital cataract patients who received phacoemulsification with IOL implantation were collected.The AMO (ZMB00) IOL was implanted in 34 eyes of 24 patients in the multifocal IOL group,and Bausch & Lomb (MI60) IOL was implanted in 33 eyes of 22 patients in the monofocal IOL group.The distance,intermediate and near vision acuity were analyzed in 3,6 and 12 months after surgery,including uncorrected visual acuity (UCVA) and best corrected visual acuity (BCVA).The wavefront aberrations,modulation transfer functions (MTF) and stereopsis were obtained with iTrace Analysis System,Optec-6500 Visual Functional Analyzer and Titmus Stereo Test Chart,respectively in 12 months after surgery.The near additional degree,removing glasses rate and myopic shift were compared between the two groups in postoperative 12 months.Results The intermediate and near UCVA in the multifocal IOL group were obviously better than those in the monofocal IOL group 3,6 and 12 months after surgery (intermediate:Z=-3.74,-4.36,-3.66;all at P=0.00.near:Z=-2.67,-2.50,-2.33;all at P<0.05).There were no significant differences between the two groups in total aberration,high and low order total aberrations,comatic aberration,trefoil aberration,spherical aberration and secondary astigmatism under the 5.0 mm optical zone in 12 months after surgery (all at P>0.05).The MTFs under the 5.0 mm optical zone and 5,10,15,20,25,30 c/d in the multifocal IOL group were insignificantly lower than those in the monofocal IOL group (all at P>0.05).In addition,the near stereopsis,near additional degree and myopic shift (AD) were reduced in the multifocal IOL group compared with monofocal IOL group (both at P<0.05).The glasses removing rate was 93.3% in the multifocal IOL group,which was significantly higher than 33.3% in the monofocal IOL group (x2 =23.25,P =0.00).No significant difference in the incidence of posterior capsular opacification was found between the two groups (P>0.05).The myopic shifting rates were 16.7% and 83.3% in the multifocal IOL group and monofocal IOL group,with a significant difference between the two groups (x2=15.02,P=0.00).Conclusions The aspheric multifical IOL implantation can achieve good and stable distance,intermediate and near visual acuities,provide better near stereopsis,reduce postoperative dependence on spectacles and decrease the incidence of myopic shift in child cataract patients.