Objective To explore the efficacy and safety of pegylated doxorubicin liposome(PLD)in neoadjuvant chemotherapy(NAC)for breast cancer.Methods A total of 126 breast cancer patients treated with the NAC regimen containing PLD in the Third Xiangya Hospital of Central South University from Janu-ary 2018 to December 2022 were selected as the research subjects.The clinical efficacy,pathological efficacy and occurrence of adverse reactions of the patients were observed,and total pathological complete response(tpCR)rates of different molecular subtypes were analyzed.The changes in left ventricular ejection fraction(LVEF)were compared between before and after treatment.Results Among 126 patients,the clinical efficacy of NAC in 119 cases was evaluated,and the objective response rate(ORR)was 67.2％(80/119).A total of 123 patients were evaluated for the pathological efficacy,breast cancer pathological complete response(bpCR)rate was 13.8％(17/123);tpCR rate was 12.2％(15/123),in which the the human epidermal growth factor receptor 2(HER2)positive[hormone receptor(HR)negative]type had the highest rate(33.3％),followed by triple-negative(21.7％).There was no hand-foot syndrome occurred,and only one case developed the grade 2 oral mucositis.LVEF showed no statistically significant difference between before and after treatment[69.9％(51.0％,87.0％)vs.69.0％(57.0％,85.0％)](P＞0.05).Conclusion The NAC regimen containing PLD has a better effect on HER2—positive and triple-negative breast cancer.

Aim To investigate the effect and mechanism of angiotensin Ⅱ(Ang Ⅱ)on ferroptosis in white adi-pocytes.Methods The 3T3-L1 preadipocytes were differentiated into white adipocytes by inducer stimulation.The experiment was divided into control group,Ang Ⅱ group,Ang Ⅱ+Fer-1(ferroptosis inhibitor)group and Ang Ⅱ+PFT-α(p53 inhibitor)group.Ang Ⅱ was used to treat cells.RT-qPCR and Western blot were used to detect the expression levels of ferroptosis factors and adipokines.JC-1 kit was used to detect mitochondrial membrane potential(MMP)level.Iron ion kit was used to detect intracellular iron content.Glutathione(GSH)kit was used to detect GSH content.Fer-1 and Ang Ⅱ were added to treat cells to detect the the changes of ferroptosis level.The expression of p53 and spermidine/spermine N1-acetyltransferase 1(SAT1)protein was detected.Subsequently,PFT-α and Ang Ⅱ were added to co-treat cells to detect the changes of p53 and SAT1 protein expression,and to observe the effect of inhibiting p53 expression on the expression levels of ferroptosis factors and adipokines.Results 3T3-L1 cells were successfully differentiated into white adipocytes by stimulator-induced differentiation.Ang Ⅱ induced ferroptosis in white adipocytes.RT-qPCR results showed that compared with control group,the mRNA expression of anti-ferroptosis factor glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11)and iron regulatory protein 1(IRP-1)was down-regulated in Ang Ⅱ group,and the mRNA expression of pro-ferroptosis factor acyl-CoA synthetase of long-chain family member 4(ACSL4)was up-regulated.Western blot results showed that compared with control group,the protein expression of SLC7A11 and GPX4 was down-regulated in Ang Ⅱ group,and the protein expression of ACSL4 was up-regulated.Ang Ⅱ treatment increased the content of intracellular iron ions and decreased the levels of GSH and MMP.Compared with Ang Ⅱ group,the mRNA expression of IRP-1 and SLC7A11 was up-regulated in Ang Ⅱ+Fer-1 group.Ang Ⅱ induced changes in the expression profile of adipokines in white adipocytes.Western blot results showed that compared with control group,the protein ex-pression of pro-inflammatory adipokine leptin(LEP),resistin(RETN),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)was up-regulated in Ang Ⅱ group,and the protein expression of anti-inflammatory adipokine adiponectin(AD-PN)and omentin 1(ITLN1)was down-regulated.In addition,Ang Ⅱ increased the protein expression of p53 and SAT1.Inhibition of p53 expression can improve the level of ferroptosis and adipokine expression in white adipocytes trea-ted with Ang Ⅱ.Western blot results showed that compared with Ang Ⅱ group,the protein expression of p53 and SAT1 was down-regulated in Ang Ⅱ+PFT-α group,the protein expression of SLC7A11 and GPX4 was up-regulated,and the protein expression of ACSL4 was down-regulated.The protein expression of ADPN was up-regulated in Ang Ⅱ+PFT-αgroup,and the protein expression of TNF-α,LEP and RETN was down-regulated.Conclusion Ang Ⅱ induces fer-roptosis in white adipocytes through activating the p53/SAT1 signaling pathway.

Background:To compare the efficacy and safety of monthly administrations of gonadotropin releasing hormone (GnRH) agonists LY01005 and Zoladex ? in Chinese patients with premenopausal breast cancer. Methods:From October 2020 to November 2021, 188 premenopausal breast cancer patients were enrolled in 34 hospitals and randomized 1:1 to receive either LY01005 or Zoladex ? every 28 days for a total of three injections. All patients concomitantly received oral tamoxifen (TAM). The primary efficacy endpoint was cumulative probability of maintaining menopausal level [oestradiol (E2) ≤30 pg/ml] from day 29 to day 85. The second efficacy endpoint included changes in E2, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) compared with the baseline. Pharmacokinetics (PK), pharmacodynamics (PD), and safety were analyzed. The study also evaluated the pharmacokinetic and pharmacodynamic characteristics of LY01005. Results:A total of 188 patients were randomised and 187 patients received either LY01005 or Zoladex ?. Cumulative probabilities of maintaining menopausal level (E2≤30 pg/ml) from day 29 to day 85 were 93.1% for LY01005 and 86.3% for Zoladex ?. The between-group difference was 6.8% (95% CI: -2.3%, 15.9%) and primary efficacy in the LY01005 group was not inferior to that in the Zoladex ? group. Changes in E2, LH, and FSH levels compared with the baseline were equivalent between the two groups (E2: 89.34% to 90.23% vs. 82.11% to 85.02%; LH: 88.89% to 95.52% vs. 89.70% to 97.02%; FSH: 75.36% to 80.85% vs.73.07% to 80.24%, respectively). After three consecutive doses of LY01005, the LH and FSH levels of the subjects showed a transient increase after the first dose, reached a peak on the second day and then started to decrease. The LH and FSH reached a lower level and remained at or below that level until the 85th day. Both treatments were well-tolerated. Conclusion:LY01005 is as effective as Zoladex ? in suppressing E2 to menopausal levels in Chinese patients with premenopausal breast cancer, with a similar safety profile.

OBJECTIVE To optimize the extraction process of organic acids from Angelica sinensis using deep eutectic solvents （DESs）， and conduct characterization， antioxidant activity evaluation， and extraction mechanism analysis. METHODS The conductor-like screening model for realistic solvation with segment activity coefficients （COSMO-SAC） was employed to screen the types of DESs. With total organic acid content as the response value， single-factor experiments and Box-Behnken response-surface methodology were used to optimize the extraction conditions. Using A. sinensis decoction pieces and/or A. sinensis methanol extract as references， scanning electron microscope and Fourier transform infrared spectrometer （FTIR） were applied to characterize the products. Additionally， the 1，1-diphenyl-2-picrylhydrazyl （DPPH） and 2，2′-azino-bis（3-ethylbenzothiazoline-6- sulfonic acid） （ABTS） free radical scavenging capacities were determined. Density functional theory （DFT） was used to analyze the extraction mechanism of ferulic acid and chlorogenic acid by the DESs. RESULTS & CONCLUSIONS The optimal DESs was choline chloride-propanediol. The optimal extraction conditions for organic acids from A. sinensis were as follows： choline chloride- propanediol molar ratio of 1∶1， DESs water content of 70%， solid-liquid ratio of 1∶10， heating temperature of 57 ℃， and heating and stirring time of 8 min. In three validation experiments， the total organic acid content was 2.92 mg/g， yielding a relative error of 0.34% compared to the predicted value （2.91 mg/g）. Compared with A. sinensis decoction pieces and methanol extracts， the agglomerated structure of the DESs extract powder almost disappeared， showing the presence of lamellar structures similar to those of the intestinal wall. Compared with the methanol extract， the DES extract exhibited higher FTIR characteristic peak intensity and peak area integration， as well as stronger scavenging capacities against DPPH and ABTS free radicals. The extraction of organic acids from A. sinensis by DESs is the result of the combined effects of polarity matching， hydrogen bonding， and structural adaptation.

Objective To investigate serum metabolites and metabolic pathways alterations in patients with ischemic stroke(IS)through metabolomic analysis,and to identify reliable serum metabolic biomarkers for IS diagnosis.Methods This prospective study enrolled patients with IS admitted to the Department of Neurology at Xiangcheng People's Hospital of Suzhou from December 1,2022 to December 31,2023.Age-and sex-matched healthy individuals were recruited as controls during the same period.Baseline characteristics were collected,including age,sex,height,body mass index,and blood pressure.Venous blood samples were obtained after an 8 h fast for biochemical analysis of blood glucose,total bilirubin,serum creatinine,urea nitrogen,total cholesterol,triglycerides,high-density lipoprotein cholesterol,and low-density lipoprotein cholesterol.Serum metabolites of both groups were extracted and analyzed using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry.Metabolomic data were processed using Simca-p software for unsupervised principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)to evaluate group separation and experimental stability.Differential metabolites were defined by variable importance in projection(VIP)≥1.0,fold change(FC)≥2.0 or ≤0.5,and P＜0.05.Drug-derived exogenous metabolites were excluded by cross-referencing the Human Metabolome Database(HMDB,https://hmdb.ca/)and PubChem(https://pubchem.ncbi.nlm.nih.gov/).MetaboAnalyst 6.0(http://www.metaboanalyst.ca),a comprehensive web-based tool for metabolomic data analysis,was employed to map differential metabolites to the Kyoto encyclopedia of genes and genomes(KEGG)databased and to perform pathway enrichment analysis.Machine learning models were developed using Python.Least absolute shrinkage and selection operator(LASSO)regression and random forest(RF)algorithms were employed to identify diagnostic biomarkers capable of effectively distinguishing IS patients from controls.Metabolites identified by both methods were integrated into an extreme gradient boosting(XGBoost)model.Model performance was evaluated using receiver operating characteristic(ROC)curves with 5-fold cross-validation and internal validation(70％training,30％validation set).Results A total of 51 IS patients and 51 matched controls were included.(1)A total of 1 255 serum metabolites were identified(964 in positive ion mode,291 in negative ion mode).PC A and OPLS-DA demonstrated distinct metabolic separation between IS patients and controls.In IS group,260 metabolites were upregulated and 337 downregulated in positive ion mode;99 were upregulated and 34downregulated in negative ion mode.(2)Among the 1 255 metabolites,259 were identified as differential metabolites based on the criteria of VIP ≥ 1.0,FC≥2.0 or≤0.5 and P＜0.05.After excluding drug-derived metabolite through referencing HMDB and PubChem databases,a total of 220 endogenous differential metabolites were found to coexist in both positive and negative ion modes.Among them,119 metabolites were up-regulated and 101 were down-regulated in the IS group.The expression of these 220 metabolites showed significant differences between the IS and control groups.(3)KEGG pathway analysis highlighted five dysregulated pathways:upregulation of denovo triacylglycerol biosynthesis,glycerophosphate shuttle,and cardiolipin biosynthesis;downregulation of bile acid biosynthesis and methylhistidine metabolism.(4)LASSO and RF algorithms identified 24 and 30 candidate biomarkers,respectively.Four overlapping metabolites were selected:2-((3R)-3-((3R,5S,7S,9S,10S,13R,14S,17R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)butanamido)ethane-1-sulfonic acid(m/z 971.571 29),arginine-conjugated cholic acid(m/z 587.379 21),laccaic acid A(m/z 576.010 93)and NCGC00380235-01_C32H48O9_beta-D-xylopyranoside,3,17-dihydroxyspirosta-5,25(27)-dien-1-yl(m/z 559.326 48).The expression levels of 2-((3R)-3-((3R,5S,7S,9S,10S,13R,14S,17R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)butanamido)ethane-1-sulfonic acid(m/z 971.571 29),arginine-conjugated cholic acid(m/z 587.379 21),and laccaic acid A(m/z 576.010 93)were upregulated,while the expression level of NCGC00380235-01_C32H48O9_beta-D-xylopyranoside,3,17-dihydroxyspirosta-5,25(27)-dien-1-yl(m/z 559.326 48)was downregulated.An IS diagnostic model was established based on four metabolic biomarkers using the XGBoost algorithm.The area under the ROC curve was 1.000(95％CI 1.000-1.000)in the training set and 0.988 in the validation set(95％CI 0.963-1.000).Conclusions Patients with IS exhibit significant metabolic disturbance.The four identified biomarkers may serve as potential biomarkers for the effective identification of IS:2-((3 R)-3-((3R,5S,7S,9S,10S,13R,14S,17R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)butanamido)ethane-1-sulfonic acid(m/z971.571 29),arginine-conjugated cholic acid(m/z587.379 21),laccaic acid A(m/z 576.010 93),and NCGC00380235-01_C32H48O9_beta-D-xylopyranoside,3,17-dihydroxyspirosta-5,25(27)-dien-1-yl(m/z 559.326 48).

Aim To investigate the effect and mechanism of angiotensin Ⅱ(Ang Ⅱ)on ferroptosis in white adi-pocytes.Methods The 3T3-L1 preadipocytes were differentiated into white adipocytes by inducer stimulation.The experiment was divided into control group,Ang Ⅱ group,Ang Ⅱ+Fer-1(ferroptosis inhibitor)group and Ang Ⅱ+PFT-α(p53 inhibitor)group.Ang Ⅱ was used to treat cells.RT-qPCR and Western blot were used to detect the expression levels of ferroptosis factors and adipokines.JC-1 kit was used to detect mitochondrial membrane potential(MMP)level.Iron ion kit was used to detect intracellular iron content.Glutathione(GSH)kit was used to detect GSH content.Fer-1 and Ang Ⅱ were added to treat cells to detect the the changes of ferroptosis level.The expression of p53 and spermidine/spermine N1-acetyltransferase 1(SAT1)protein was detected.Subsequently,PFT-α and Ang Ⅱ were added to co-treat cells to detect the changes of p53 and SAT1 protein expression,and to observe the effect of inhibiting p53 expression on the expression levels of ferroptosis factors and adipokines.Results 3T3-L1 cells were successfully differentiated into white adipocytes by stimulator-induced differentiation.Ang Ⅱ induced ferroptosis in white adipocytes.RT-qPCR results showed that compared with control group,the mRNA expression of anti-ferroptosis factor glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11)and iron regulatory protein 1(IRP-1)was down-regulated in Ang Ⅱ group,and the mRNA expression of pro-ferroptosis factor acyl-CoA synthetase of long-chain family member 4(ACSL4)was up-regulated.Western blot results showed that compared with control group,the protein expression of SLC7A11 and GPX4 was down-regulated in Ang Ⅱ group,and the protein expression of ACSL4 was up-regulated.Ang Ⅱ treatment increased the content of intracellular iron ions and decreased the levels of GSH and MMP.Compared with Ang Ⅱ group,the mRNA expression of IRP-1 and SLC7A11 was up-regulated in Ang Ⅱ+Fer-1 group.Ang Ⅱ induced changes in the expression profile of adipokines in white adipocytes.Western blot results showed that compared with control group,the protein ex-pression of pro-inflammatory adipokine leptin(LEP),resistin(RETN),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)was up-regulated in Ang Ⅱ group,and the protein expression of anti-inflammatory adipokine adiponectin(AD-PN)and omentin 1(ITLN1)was down-regulated.In addition,Ang Ⅱ increased the protein expression of p53 and SAT1.Inhibition of p53 expression can improve the level of ferroptosis and adipokine expression in white adipocytes trea-ted with Ang Ⅱ.Western blot results showed that compared with Ang Ⅱ group,the protein expression of p53 and SAT1 was down-regulated in Ang Ⅱ+PFT-α group,the protein expression of SLC7A11 and GPX4 was up-regulated,and the protein expression of ACSL4 was down-regulated.The protein expression of ADPN was up-regulated in Ang Ⅱ+PFT-αgroup,and the protein expression of TNF-α,LEP and RETN was down-regulated.Conclusion Ang Ⅱ induces fer-roptosis in white adipocytes through activating the p53/SAT1 signaling pathway.

Objective To investigate serum metabolites and metabolic pathways alterations in patients with ischemic stroke(IS)through metabolomic analysis,and to identify reliable serum metabolic biomarkers for IS diagnosis.Methods This prospective study enrolled patients with IS admitted to the Department of Neurology at Xiangcheng People's Hospital of Suzhou from December 1,2022 to December 31,2023.Age-and sex-matched healthy individuals were recruited as controls during the same period.Baseline characteristics were collected,including age,sex,height,body mass index,and blood pressure.Venous blood samples were obtained after an 8 h fast for biochemical analysis of blood glucose,total bilirubin,serum creatinine,urea nitrogen,total cholesterol,triglycerides,high-density lipoprotein cholesterol,and low-density lipoprotein cholesterol.Serum metabolites of both groups were extracted and analyzed using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry.Metabolomic data were processed using Simca-p software for unsupervised principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)to evaluate group separation and experimental stability.Differential metabolites were defined by variable importance in projection(VIP)≥1.0,fold change(FC)≥2.0 or ≤0.5,and P＜0.05.Drug-derived exogenous metabolites were excluded by cross-referencing the Human Metabolome Database(HMDB,https://hmdb.ca/)and PubChem(https://pubchem.ncbi.nlm.nih.gov/).MetaboAnalyst 6.0(http://www.metaboanalyst.ca),a comprehensive web-based tool for metabolomic data analysis,was employed to map differential metabolites to the Kyoto encyclopedia of genes and genomes(KEGG)databased and to perform pathway enrichment analysis.Machine learning models were developed using Python.Least absolute shrinkage and selection operator(LASSO)regression and random forest(RF)algorithms were employed to identify diagnostic biomarkers capable of effectively distinguishing IS patients from controls.Metabolites identified by both methods were integrated into an extreme gradient boosting(XGBoost)model.Model performance was evaluated using receiver operating characteristic(ROC)curves with 5-fold cross-validation and internal validation(70％training,30％validation set).Results A total of 51 IS patients and 51 matched controls were included.(1)A total of 1 255 serum metabolites were identified(964 in positive ion mode,291 in negative ion mode).PC A and OPLS-DA demonstrated distinct metabolic separation between IS patients and controls.In IS group,260 metabolites were upregulated and 337 downregulated in positive ion mode;99 were upregulated and 34downregulated in negative ion mode.(2)Among the 1 255 metabolites,259 were identified as differential metabolites based on the criteria of VIP ≥ 1.0,FC≥2.0 or≤0.5 and P＜0.05.After excluding drug-derived metabolite through referencing HMDB and PubChem databases,a total of 220 endogenous differential metabolites were found to coexist in both positive and negative ion modes.Among them,119 metabolites were up-regulated and 101 were down-regulated in the IS group.The expression of these 220 metabolites showed significant differences between the IS and control groups.(3)KEGG pathway analysis highlighted five dysregulated pathways:upregulation of denovo triacylglycerol biosynthesis,glycerophosphate shuttle,and cardiolipin biosynthesis;downregulation of bile acid biosynthesis and methylhistidine metabolism.(4)LASSO and RF algorithms identified 24 and 30 candidate biomarkers,respectively.Four overlapping metabolites were selected:2-((3R)-3-((3R,5S,7S,9S,10S,13R,14S,17R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)butanamido)ethane-1-sulfonic acid(m/z 971.571 29),arginine-conjugated cholic acid(m/z 587.379 21),laccaic acid A(m/z 576.010 93)and NCGC00380235-01_C32H48O9_beta-D-xylopyranoside,3,17-dihydroxyspirosta-5,25(27)-dien-1-yl(m/z 559.326 48).The expression levels of 2-((3R)-3-((3R,5S,7S,9S,10S,13R,14S,17R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)butanamido)ethane-1-sulfonic acid(m/z 971.571 29),arginine-conjugated cholic acid(m/z 587.379 21),and laccaic acid A(m/z 576.010 93)were upregulated,while the expression level of NCGC00380235-01_C32H48O9_beta-D-xylopyranoside,3,17-dihydroxyspirosta-5,25(27)-dien-1-yl(m/z 559.326 48)was downregulated.An IS diagnostic model was established based on four metabolic biomarkers using the XGBoost algorithm.The area under the ROC curve was 1.000(95％CI 1.000-1.000)in the training set and 0.988 in the validation set(95％CI 0.963-1.000).Conclusions Patients with IS exhibit significant metabolic disturbance.The four identified biomarkers may serve as potential biomarkers for the effective identification of IS:2-((3 R)-3-((3R,5S,7S,9S,10S,13R,14S,17R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)butanamido)ethane-1-sulfonic acid(m/z971.571 29),arginine-conjugated cholic acid(m/z587.379 21),laccaic acid A(m/z 576.010 93),and NCGC00380235-01_C32H48O9_beta-D-xylopyranoside,3,17-dihydroxyspirosta-5,25(27)-dien-1-yl(m/z 559.326 48).

Background:To compare the efficacy and safety of monthly administrations of gonadotropin releasing hormone (GnRH) agonists LY01005 and Zoladex ? in Chinese patients with premenopausal breast cancer. Methods:From October 2020 to November 2021, 188 premenopausal breast cancer patients were enrolled in 34 hospitals and randomized 1:1 to receive either LY01005 or Zoladex ? every 28 days for a total of three injections. All patients concomitantly received oral tamoxifen (TAM). The primary efficacy endpoint was cumulative probability of maintaining menopausal level [oestradiol (E2) ≤30 pg/ml] from day 29 to day 85. The second efficacy endpoint included changes in E2, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) compared with the baseline. Pharmacokinetics (PK), pharmacodynamics (PD), and safety were analyzed. The study also evaluated the pharmacokinetic and pharmacodynamic characteristics of LY01005. Results:A total of 188 patients were randomised and 187 patients received either LY01005 or Zoladex ?. Cumulative probabilities of maintaining menopausal level (E2≤30 pg/ml) from day 29 to day 85 were 93.1% for LY01005 and 86.3% for Zoladex ?. The between-group difference was 6.8% (95% CI: -2.3%, 15.9%) and primary efficacy in the LY01005 group was not inferior to that in the Zoladex ? group. Changes in E2, LH, and FSH levels compared with the baseline were equivalent between the two groups (E2: 89.34% to 90.23% vs. 82.11% to 85.02%; LH: 88.89% to 95.52% vs. 89.70% to 97.02%; FSH: 75.36% to 80.85% vs.73.07% to 80.24%, respectively). After three consecutive doses of LY01005, the LH and FSH levels of the subjects showed a transient increase after the first dose, reached a peak on the second day and then started to decrease. The LH and FSH reached a lower level and remained at or below that level until the 85th day. Both treatments were well-tolerated. Conclusion:LY01005 is as effective as Zoladex ? in suppressing E2 to menopausal levels in Chinese patients with premenopausal breast cancer, with a similar safety profile.

AIM: To analyze changes in the ocular surface parameters of keratoconus after long-term wearing of rigid gas permeable contact lens(RGPCL).METHODS:Prospective case study. A total of 113 keratoconus patients(213 eyes)fitted with RGPCL in the optometry center of Gansu Provincial Hospital from January 2018 to January 2022 were included. They were divided into three groups according to the severity of keratoconus, including 42 cases(80 eyes)in mild keratoconus group, 54 cases(102 eyes)in moderate keratoconus group and 17 cases(31 eyes)in severe keratoconus group. Furthermore, the non-invasive tear break-up time(NIBUT), non-invasive tear meniscus height(NITMH), red eye index, lipid layer thickness, fluorescent corneal staining, meibomian gland secretory function, Schirmer I test and ocular surface disease index(OSDI)scores were observed by Keratograph analyzer before and after wearing RGPCL for 1 wk, 1, 3, 6, 12 mo, respectively.RESULTS: There were no statistical significance in the age, NIBUT, NITMH, lipid layer thickness, meibomian gland secretory function and Schirmer I test among the three groups(P>0.05), while there were statistical significance in the sphere, cylinder, spherical equivalent, best corrected visual acuity(BCVA), non-contact intraocular pressure(IOPNCT), anterior, posterior corneal surface Kmax, corneal surface thickness at the thinnest point, eye redness index, fluorescent corneal staining, and OSDI(P<0.05). In the mild keratoconus group, NIBUT had statistical differences at 3, 6 and 12 mo after wearing RGPCL(P<0.05), NITMH had statistical differences in 6 and 12 mo(P<0.05), the eye redness index, fluorescent corneal staining and OSDI scores had statistical differences in 1 wk and 1 mo(P<0.05), and lipid layer thickness and meibomian gland secretory function had statistical differences in 12 mo(P<0.05). In the moderate keratoconus group, there were statistical differences in NIBUT at 6 and 12 mo after wearing lenses(P<0.05); there were statistical differences in the NITMH, lipid layer thickness and meibomian secretory function at 12 mo after wearing lens(P<0.05); there were statistical differences in the eye redness index at 1 wk, 1 and 3 mo after wearing RGPCL(P<0.05); there were statistical differences in the fluorescent corneal staining at 1 wk after wearing RGPCL(P<0.05); there were statistical differences in the OSDI at 1 wk and 1 mo after wearing RGPCL(P<0.05). In the severe keratoconus group, there were statistical differences in the NIBUT, NITMH and eye redness index at 1 wk, 1, 3, 6 and 12 mo after wearing RGPCL(P<0.05); there were statistical differences in the lipid layer thickness at 6,12 mo after wearing RGPCL(P<0.05); there were statistical differences in the fluorescent corneal staining and OSDI scores at 1 wk, 6 and 12 mo after wearing RGPCL(P<0.05); there were statistical differences in the meibomian secretory function at 6 and 12 mo after wearing RGPCL(P<0.05); and there were statistical differences in the Schirmer I test at 12 mo after wearing RGPCL(P<0.05).CONCLUSION: Long-term wearing of RGPCL can affect the ocular surface microcirculation in keratoconus patients, thus making differences in patients subjective. However, it has no significant impact on the visual quality of patients. Therefore, long-term wearing of RGPCL is safely to control the progression of keratoconus.

Objective To investigate the effects of icariin on high glucose-induced autophagy and apoptosis of podocytes,and the regulating effects on mammalian target of rapamycin(mTOR)/serine-threonine kinase(Akt)/cyclic adenosine monophosphate response element binding protein(CREB)pathway.Methods The mouse podocytes MPC5 were taken and divided into five groups:normal control group(5.5 mmol·L-1 glucose),high glucose group(30 mmol·L-1 glucose),icariin group(30 mmol·L-1glucose+5 μmol·L-1icariin),GDC-0349 group(30 mmol·L-1glucose+50 μmol·L-1 GDC-0349),icariin+GDC-0349 group(30 mmol·L-1 glucose+5 μmol·L-1 icariin+50 μmol·L-1 GDC-0349).Cultured for 48 hours,the tetramethylazozolium salt method was used to detect the viability of MPC5 cells;acridine orange staining was used to observe the autophagy of MPC5 cells;apoptosis of MPC5 cells was detected by flow cytometry;Western blotting was used to detect the expression of autophagy[microtubule associated protein one light chain 3(LC3)II,LC3Ⅰ,autophagy-related protein(Beclin-1)],apoptosis[Bcl-2 related X protein(Bax),B cell lymphoma-2(Bcl-2)]and mTOR/Akt/CREB pathway-related proteins of MPC5 cells.Results Compared with the normal control group,the cell viability,expression levels of Bcl-2,phosphorylated mTOR(p-mTOR)/mTOR,phosphorylated Akt(p-Akt)/Akt,phosphorylated CREB(p-CREB)/CREB protein of MPC5 cells in the high glucose group were significantly decreased(P＜0.05),the autophagy ability was enhanced,the autophagosome showed orange fluorescence,and the apoptosis rate,LC3Ⅱ/LC3Ⅰ,Beclin-1,Bax protein expression levels were significantly increased(P＜0.05).Compared with the high glucose group,the cell viability,LC3Ⅱ/LC3Ⅰ,Beclin-1,Bcl-2,p-mTOR/mTOR,p-Akt/Akt,p-CREB/CREB protein expression levels of MPC5 cells in icariin group were significantly increased,the autophagy ability was further enhanced,the number of autophagosomes was increased,the autophagosomes showed brick red fluorescence(P＜0.05),the apoptosis rate and Bax protein expression level were significantly decreased(P＜0.05),and the cell viability,LC3Ⅱ/LC3Ⅰ,Beclin-1,Bcl-2,p-mTOR/mTOR,p-Akt/Akt and p-CREB/CREB proteins expression levels of MPC5 cells in GDC-0349 group were significantly decreased,the autophagy ability was weakened,the number of autophagosomes was reduced,the autophagosomes showed orange fluorescence(P＜0.05),and the apoptosis rate and Bax protein expression level were significantly increased(P＜0.05);icariin+GDC-0349 could reverse the effect of icariin on high glucose induced MPC5 cells(P＜0.05).Conclusion Icariin promotes elevated glucose-induced podocyte autophagy and inhibits apoptosis by activating the mTOR/Akt/CREB pathway.