Objective To investigate the correlation between serum vascular endothelial growth factor(VEGF)level and the development and severity of diabetes mellitus cystoid macular edema(CME).Methods A total of 57 patients(57 eyes)with diabetic CME(the case group)and diabetes without eye complications(the control group)admitted to the Vitreoretinal Surgery Department of Tangshan Eye Hospital from June 2023 to June 2024 were prospectively selected,and all of them underwent systematic ophthalmic specialty examination and serum VEGF detection.Multivariate Logistic regression analysis was used to identify the risk factors of diabetes mellitus CME.The diagnostic value of VEGF in patients with diabetes CME was analyzed by receiver operating characteristic(ROC)curve.Spearman correlation was used to analyze the relationship between VEGF levels and the severity of CME in patients with diabetes,such as corrected visual,number of cystoid edema and non-round index of macular foveal avascular zone(FAZ).Results Compared with the control group,the course of diabetes in the case group was longer,the incidence of hypertension was higher,levels of glycosylated hemoglobin(HbA1c),serum creatinine and serum VEGF were higher,and the estimated glomerular filtration rate(eGFR)was lower(P＜0.05).Multivariate Logistic regression results showed that long duration of diabetes,increased levels of HbA1c and VEGF were risk factors for CME in diabetes mellitus patients,while elevated eGFR was protective factor(P＜0.05).ROC curve analysis showed that the area under the curve(AUC)of VEGF in diagnosing diabetes mellitus CME was 0.758(95%CI:0.669-0.834).The AUC of the combined application of diabetes duration,HbA1c,eGFR and VEGF in the diagnosis of diabetes mellitus CME was 0.916(95%CI:0.877-0.941),which was significantly higher than that of the application of VEGF alone(P＜0.05).The VEGF level was negatively correlated with the corrected visual acuity[0.34(0.24,0.44)]in patients with diabetic CME,and positively correlated with the detection of cystoid macular edema[3(2,5)]and FAZ non-roundness index[1.16(1.08,1.20)](rs were 0.771,0.700,respectively,P＜0.05).Conclusion Serum VEGF levels are closely related to the onset and severity of CME,and which can be used as a reliable reference index for the diagnosis of CME.

Objective To investigate the correlation between serum vascular endothelial growth factor(VEGF)level and the development and severity of diabetes mellitus cystoid macular edema(CME).Methods A total of 57 patients(57 eyes)with diabetic CME(the case group)and diabetes without eye complications(the control group)admitted to the Vitreoretinal Surgery Department of Tangshan Eye Hospital from June 2023 to June 2024 were prospectively selected,and all of them underwent systematic ophthalmic specialty examination and serum VEGF detection.Multivariate Logistic regression analysis was used to identify the risk factors of diabetes mellitus CME.The diagnostic value of VEGF in patients with diabetes CME was analyzed by receiver operating characteristic(ROC)curve.Spearman correlation was used to analyze the relationship between VEGF levels and the severity of CME in patients with diabetes,such as corrected visual,number of cystoid edema and non-round index of macular foveal avascular zone(FAZ).Results Compared with the control group,the course of diabetes in the case group was longer,the incidence of hypertension was higher,levels of glycosylated hemoglobin(HbA1c),serum creatinine and serum VEGF were higher,and the estimated glomerular filtration rate(eGFR)was lower(P＜0.05).Multivariate Logistic regression results showed that long duration of diabetes,increased levels of HbA1c and VEGF were risk factors for CME in diabetes mellitus patients,while elevated eGFR was protective factor(P＜0.05).ROC curve analysis showed that the area under the curve(AUC)of VEGF in diagnosing diabetes mellitus CME was 0.758(95%CI:0.669-0.834).The AUC of the combined application of diabetes duration,HbA1c,eGFR and VEGF in the diagnosis of diabetes mellitus CME was 0.916(95%CI:0.877-0.941),which was significantly higher than that of the application of VEGF alone(P＜0.05).The VEGF level was negatively correlated with the corrected visual acuity[0.34(0.24,0.44)]in patients with diabetic CME,and positively correlated with the detection of cystoid macular edema[3(2,5)]and FAZ non-roundness index[1.16(1.08,1.20)](rs were 0.771,0.700,respectively,P＜0.05).Conclusion Serum VEGF levels are closely related to the onset and severity of CME,and which can be used as a reliable reference index for the diagnosis of CME.

Objective To investigate the effect of decitabine (DAC) on human acute myeloid leukemia (AML) cell line HL-60 and the regulating of natural killer (NK) cell activating receptor (NKG2D) ligands(NKG2DL), and to detect the molecular mechanism of JAK-STAT3-SOCS signaling pathway. Methods The effect of DAC on the proliferation of HL-60 was detected by using CCK-8 assay. The cell apoptosis was analyzed by using Annexin-V/PI double standard method. The expressions of receptor NKG2DL including MICA/B and ULBPs in HL-60 cells were detected by using flow cytometry (FCM). The killing activity of NK cells was analyzed by using carboxy fluorescein diacetate succinimidyl ester (CFSE). The expressions of JAK/STAT3 signaling pathway or molecules including STAT3, its upstream kinases JAK1, JAK2 and the negative regulator of STAT3,SOCS-1,SOCS-3 were examined by Western blot.Methylation level of the SOCS-1,SOCS-3 gene after the treatment of DAC was analyzed by using methylation-sensitive high resolution melting(MS-HRM). Results There was an obvious inhibitory effect of DAC on HL-60 cells. The cell viability of HL-60 treated with 0.2, 0.5, and 1.0 μmol/L DAC for 48 h was decreased by (25±11) %, (39±8) % and (50±7)%(P<0.01)respectively compared with those cells without DAC treatment.The incidence of apoptosis was (24.77±7.50) %, (27.10±4.48) % and (30.53±3.93) % after DAC treatment for 48h respectively, which were higher than that of untreated cells[(3.11±0.50)%](P<0.01).DAC induced a significant up-regulation of MICA/B, ULBP-1, ULBP-3 in HL-60 cells, and enhanced the sensitivity of HL-60 cells to NK cytotoxicity. Western blot results showed that a down-regulating expression of STAT3 and JAK1, JAK2 protein was detected, in addition to the phosphor-STAT3 and phosphor-JAKs in HL-60 cells after DAC treatment, but the expressions of SOCS-1 and SOCS-3 protein were increased. HRM results showed that DAC could inhibit the methylation of SOCS-3 gene. Conclusion DAC can inhibit the proliferation of HL-60 cells, upregulate the expression of NKG2DL and enhance the cytotoxicity of NK targeted to HL-60 cells, which might be related to the activity regulation of intracellular JAK-STAT3-SOCS signaling pathway.

OBJECTIVETo investigate the molecular mechanism of the growth inhibitory effect of matrine on K562 cells in JAK/STAT3 mediated signal pathway.

METHODSWestern blot analyses were performed to investigate the differential expression of JAK2, STAT3, phosphor-STAT3 (Tyr705 & Ser727) and phosphor-JAK2 proteins after matrine treatment in K562 cells with or without human recombinant interleukin 6 (IL-6) pretreatment. The expression of STAT3 response gene products such as Bcl-xL, Cyclin D1 and c-Myc, were investigated by Western blot and quantitative real time RT-PCR (qRT-PCR). Expression of IL-6, a potent upstream activating factor of JAK/STAT3 pathway, was analyzed by both real time qRT-PCR and ELISA.

RESUTLSWestern blot revealed that matrine treatment resulted in a strong down-regulation of phosphor-STAT3 both in Tyr705 and Ser727 sites or phosphor-JAK2 proteins expression without significant effects on the total STAT3 and JAK2 proteins. The expression of phosphor-Tyr705 STAT3 and phosphor-Ser727 STAT3 was decreased to 0.370 ± 0.172 in K562 cells treated with 0.5 mg/ml matrine for 48 h, respectively, from 0.690 ± 0.119 and 1.150 ± 0.263 in control cells, accompanied with a dramatical down-regulation of phosphor-JAK2 from 0.670 ± 0.137 to 0.049 ± 0.057 (P<0.05). In addition, it was found that the expression of Bcl-xL, Cyclin D1, c-Myc was decreased both at the transcription and protein level in K562 cells after matrine treatment. Matrine treatment resulted in a significant decrease in the expression level of IL-6 in K562 cells from (35.1 ± 1.93) to (10.74 ± 1.83) and (8.66 ± 1.24) pg/ml at the dose of 0.5 and 0.8 mg/ml, respectively (p<0.05). Matrine treatment could diminish the up-regulation of STAT3, JAK2, phosphor-STAT3 and phosphor-JAK2 protein following pretreatment with IL-6 in K562 cells.

CONCLUSIONMatrine exerts its anti-leukemia effect by interfering with the JAK2/STAT3 signaling pathway. The inhibition of IL-6 expression may play a pivotal role in the disruption of JAK/STAT pathway by matrine.

Alkaloids ; Down-Regulation ; Humans ; Interleukin-6 ; Janus Kinase 2 ; K562 Cells ; Quinolizines ; STAT3 Transcription Factor ; Signal Transduction ; Up-Regulation

Objective To investigate the mechanism of matrine in inhibition of proliferation the proliferation of human chronic myeloid leukemia (CML) K562 cells via MEK-ERK signaling pathway. Methods Western blot was used to detect the expression of MEK1, ERK1/2, Shc and SHP2 (the signal effect molecules of MEK-ERK pathway) in K562 cells. The transcription and translation of bcr-abl and target protein (bcl-xL, Cyclin D1, c-myc and p27) were detected by RT-PCR and Western blot. Results Matrine was able to significantly inhibit the phosphorylation of MEK1, ERK1/2, Shc and SHP2 in K562 cells and suppress the protein and mRNA expression of bcr-abl. Moreover, the expressions of bcl-xL, Cyclin D1 and c-myc were down-regulated significantly, while the expression level of p27 (a negative regulator of cell cycle progression) was increased markedly after matrine treatment. Conclusions Suppression of the growth of human CML K562 cells is related to the inhibition of bcr-abl-mediated MEK-ERK pathway activity. The down-regulation of phosphorylated proteins or protein kinases activity in signaling pathways might be an important molecular mechanism in control the activity of MEK-ERK pathway.

Objective To observe the clinical therapeutic effects of radial optic neurotomy (RON) for central retinal vein occlusion (CRVO). Methods The clinical data of 12 patients (12 eyes) with CRVO who had undergone RON were retrospectively analyzed. The patients were examined by visual acuity and fundus examination, photography, fundus fluorescein angiography, and optical coherence tomography (OCT). The visual acuities of all of the 12 eyes with severe hemorrhage and retinal edema were less than 0.1. A microvitreoretinal blade or CRVO knife was penetrate into the nasal side of the optic disk, and the incision reached the cribriform plate and even the back, and then dissected at the nasal side of the cribriform plate. The postoperative follow-up period lasted 2 to 15 months with the mean of 6.5 months, and the visual acuity, the condition of visual field and ocular fundus of the eyes undergone the surgery were observed. Results In 12 eyes undergone RON successfully, the visual acuity after the surgery improved more or less in 9 (75%), and more than or up to 0.1 in 6. The results of the examinations showed that the retinal edema and hemorrhage disappeared gradually, varicosity mitigated, retinal serous detachment mitigated or disappeared, and macular cystoid edema faded; macular fovea recovered in 3 eyes, optic nerve had a hemorrhage during the operation in 3 eyes, and the dissected part of optic nerve became atrophic afterward in 4. Conclusions RON performed at the edge of the nasal side of the optic disc is safe and effective, which may improve the visual acuity of the eyes with CRVO and mitigate retinal hemorrhage and edema.