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AIM:To study the regulation of endoplasmic reticulum stress(ERS)using the glucose regulated protein 78(GRP78)/CCAAT/enhancer binding protein homologous protein(CHOP)pathway and explore the related mech-anism of Wenyang-Shengji ointment in facilitating the repair of diabetic refractory wounds.METHODS:To establish a rat model of diabetic refractory wound repair,Sprague-Dawley(SD)rats were fed a high-fat diet and intraperitoneally in-jected with streptozotocin.Subsequently,full-thickness skin defects were induced in the dorsal region of the rats.The ex-periment included 4 groups:normal,model(diabetic refractory wounds),Wenyang-Shengji ointment,and Beifuxin(re-combinant bovine basic fibroblast growth factor gel)groups.The normal and model groups were treated with normal saline after disinfection.In the Wenyang-Shengji ointment and Beifuxin groups,the wounds were topically treated with the re-spective ointments once daily.After 14 d of treatment,wound healing was assessed and quantified using the wound healing rate.Hematoxylin-eosin(HE)staining was employed to examine the micromorphology of the wound tissue.Western blot analysis was performed to measure GRP78,CHOP and caspase-12 levels in the wound tissue.Immunohistochemical analy-sis was used to detect the expression and distribution patterns of GRP78,CHOP and caspase-12 in the wounds.Transmis-sion electron microscopy was used to observe reticulum numbers and swelling.Enzyme-linked immunosorbent assay was used to determine interleukin-1β(IL-1β)level as a pro-inflammatory factor within the wound.RESULTS:Indexes of each group were assessed 14 d after the corresponding intervention.Compared with normal group,the rats in model group exhibited a significant decrease in the wound healing rate(P＜0.01),accompanied by increased inflammatory exudation and poor granulation tissue growth.Additionally,there were increases in the expression levels of GRP78,CHOP and cas-pase-12 proteins(P＜0.01),as well as a significant elevation in the content of inflammatory factor IL-1β(P＜0.01).In contrast,compared with model group,treatment with Wenyang-Shengji ointment resulted in a significant improvement in wound healing rate(P＜0.01),reduction in inflammatory exudation,and enhanced granulation tissue growth(P＜0.01).Furthermore,there was a notable decrease in the protein expression of GRP78/CHOP/caspase-12 within the wound tissue following treatment with Wenyang-Shengji ointment(P＜0.01).The levels of inflammatory factor IL-1β also showed a sig-nificant decrease(P＜0.01).CONCLUSION:Wenyang-Shengji promotes the healing of diabetic refractory wounds,which may be associated with the downregulation of the GRP78/CHOP pathway,inhibition of excessive ERS,and reduc-tion in the level of wound cell apoptosis.
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Objective To observe the effects of Gegen Qinlian Decoction on pancreatic endoplasmic reticulum stress in mice with type 2 diabetes mellitus(T2DM);To explore its mechanism of action in the treatment of T2DM.Methods Totally 75 SPF male db/db mice were randomly divided into model group,metformin group,and Gegen Qinlian Decoction high-,medium-,low-dosage groups,with 15 mice in each group.15 db/m mice were set as the blank group.The administration groups received corresponding medicine for gavage for 12 weeks.Body mass,fasting blood glucose(FBG)and glycated hemoglobin(HbA1c)in mice were detected,HE staining was used to observe the pathological changes of pancreatic tissue,the apoptosis of islet cells was determined by TUNEL staining,Western blot was used to detect pancreatic tissue glucose regulatory protein 78(GRP78),protein kinase R-like endoplasmic reticulum kinase(PERK),p-PERK,activated transcription factor 4(ATF4)and C/EBP homologous protein(CHOP)protein expression,RT-PCR was used to detect pancreatic tissue PERK,ATF4,CHOP mRNA expressions.Results Compared with the blank group,the body mass,FBG and HbA1c contents in the model group significantly increased(P<0.01);the pancreatic tissue structure was incomplete,with blurry boundaries and vacuoles inside,leading to a significant increase in pancreatic islet cells apoptosis(P<0.01);the expressions of GRP78,p-PERK,ATF4,and CHOP proteins in pancreatic tissue significantly increased(P<0.01),and the mRNA expressions of PERK,ATF4 and CHOP significantly increased(P<0.01).Compared with the model group,the body mass,FBG and HbA1c contents of mice in each administration group significantly decreased(P<0.05,P<0.01);pathological changes in pancreatic tissue was reduced,and islet cells apoptosis decreased to varying degrees(P<0.05,P<0.01);the expressions of GRP78,p-PERK,ATF4 and CHOP proteins in pancreatic tissue significantly decreased(P<0.01)in Gegen Qinlian Decoction high-and medium-dosage groups and the metformin group,and the expressions of PERK,ATF4 and CHOP mRNA significantly decreased(P<0.05,P<0.01).Conclusion Gegen Qinlian Decoction may decreased pancreatic islet cells apoptosis,protect pancreatic cell function,and delay the progression of T2DM by inhibiting the endoplasmic reticulum stress PERK/ATF4/CHOP signaling pathway,and down-regulating the expressions of related genes and proteins.
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@#Objective To evaluate the protective effect of the activator of silent information regulator 2-related enzymes 1(SIRT1),SRT1720,on liver injury induced by acetaminophen(APAP)in mice and explore its mechanism. Methods Forty male C57BL/6J mice were randomly divided into normal control group,SRT1720 treatment group,APAP treatment group,and APAP + SRT1720 treatment group,with 10 mice in each group. Mice in SRT1720 and APAP + SRT1720 groups were given SRT1720(30 mg/kg body mass)by intragastric administration,while normal saline of equal volume was given by intragastric administration in control and APAP groups,once a day for 5 days;On the 6th day,mice in APAP and APAP + SRT1720 groups were injected i. p. with APAP(325 mg/kg body mass),while those in control and SRT1720 groups with equal volume of normal saline. After 24 hours,the peripheral blood was taken and the serum was separated,which were detected for the levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)by the corresponding kits;The liver tissue of mice was taken aseptically,observed for the pathological changes by HE staining,detected for the mRNA transcription levels of GRP78,PERK,eIF2 α,ATF4 and CHOP genes related to PERK-eIF2α-CHOP signaling pathway by RT-qPCR and detected for the relative expression levels of these corresponding proteins and Caspase12 protein by Western blot. Results Compared with normal control group,the serum ALT and AST levels of mice in APAP group significantly increased(t = 55. 21 and34. 29 respectively,each P < 0. 01);significant necrosis of hepatocytes was observed in liver tissue,the structure of hepatic lobules changed significantly,and the swelling and deformation of hepatocytes in some areas were serious;the mRNA transcription and relative protein expression levels of GRP78,PERK,eIFα,ATF4 and CHOP genes increased significantly(t = 9. 85～33. 89,each P < 0. 05)and the relative expression level of Caspase12 protein increased significantly(t = 11. 78,P < 0. 01). Compared with APAP group,the serum ALT and AST levels of mice in APAP + SRT1720 group decreased significantly(t = 42. 92 and 18. 02 respectively,each P < 0. 01);the degree of hepatocyte injury was obviously reduced and the number of swollen and deformed cells also significantly decreased;the mRNA transcription and relative protein expression levels of GRP78,PERK,eIF2α,ATF4 and CHOP decreased significantly(t = 6. 19～22. 43,each P < 0. 05)and the expression level of Caspase12 protein showed no significant decrease(t = 0. 34,P > 0. 05). Conclusion SRT1720improved APAP-induced liver injury in mice,possibly by inhibiting PERK-eIF2α-CHOP signaling pathway in endoplasmic reticulum stress(ERS).
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The biosynthesis and maturation of proteins are primarily regulated by the endoplasmic reticulum in its physiological state. Thus, the disruption of physiological homeostasis initiates the buildup of unfolded and misfolded proteins in the endoplasmic reticulum, resulting in endoplasmic reticulum stress (ERS) and unfolded protein response (UPR). One of the important pathways by which UPR maintains intracellular homeostasis under ERS is activating protein kinase R-like endoplasmic reticulum kinase (PERK). The activation of the PERK pathway stimulates eukaryotic translation initiation factor 2 subunit-α (eIF2α) phosphorylation and the selective translation of active transcription factor 4 (ATF4), and PERK induces cell apoptosis by directly binding to the promoter of pro-apoptotic transcription factor C/EBP homologous protein (CHOP). This signaling pathway is also one of the important mechanisms by which UPR participates in the regulation of hematological malignancies and immune cells in a tumor microenvironment. This article provides an overview of advancements in research into the PERK-eIF2α-ATF4-CHOP signaling pathway in hematological malignancies and the potential therapeutic benefits of targeting this signaling pathway.
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@#High-Grade B-Cell Lymphoma (HGBCL) with gene rearrangements in MYC and BCL2 and/or BCL6 is an aggressive malignancy usually presenting in advanced stages. Current recommendations suggest the use of regimens more intensive than R-CHOP (rituximab, cyclophosphamide, vincristine, doxorubicin, prednisone), which are based on retrospective studies and single-arm prospective trials that included patients who are mostly in the advanced stage, and did not receive consolidation radiotherapy. The optimal approach and treatment of HGBCL, whether limited-stage (LS) or advanced-stage, remains to be determined. Here we describe the promising outcomes of three patients with LS and low IPI HGBCL with the use of R-CHOP as induction chemotherapy regimen, which was followed by consolidation radiotherapy. Three women, 54-, 60-, and 64-years of age diagnosed to have HGBCL with MYC, and BCL2 and/or BCL6 rearrangements, with Ann Arbor stages I-IIE were included in this case series. All three patients had complete metabolic response to 6 cycles of R-CHOP and was subsequently treated with consolidation involved site radiotherapy (ISRT; total dose 30-36 Gy). Chemotherapy and radiotherapy were tolerated very well. All patients remain to be in remission, with the longest being at 23 months. Outcomes of patients with HGBCL generally remain to be poor, but this may not be the case for patients with limited-stage disease and favorable clinicopathologic risk profile. Nevertheless, the treatment of HGBCL is currently evolving and more studies are needed to determine the ideal approach and preferred chemotherapy regimen. Also, more studies are needed to elucidate the potential role of consolidation radiotherapy in patients with limited-stage HGBCL to improve survival outcomes. Findings of this case series suggest that patients with LS HGBCL may still derive benefit from R-CHOP followed by consolidation ISRT, but prospective trials are needed to confirm this.
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@#Objective: To explore the efficacy and potential mechanisms of the ethanol extract of Abelmoschus manihot (L.) Medic in contrast-induced nephropathy (CIN). Methods: CIN rat models and human renal proximal tubular cells (HK-2) with iopromide-induced injury were employed to mimic CIN conditions. The effect of Abelmoschus manihot extract on the rat models and HK-2 cells was evaluated. In rat models, kidney function, histology, oxidative stress and apoptosis were determined. In HK-2 cells, cell viability, apoptosis, mitochondrial membrane potential, and endoplasmic reticulum stress were assessed. Results: Abelmoschus manihot extract significantly improved structural and functional impairments in the kidneys of CIN rats. Additionally, the extract effectively mitigated the decline in cellular viability and reduced iopromide-induced oxidative stress and lipid peroxidation. Mechanistic investigations revealed that Abelmoschus manihot extract prominently attenuated acute endoplasmic reticulum stress-mediated apoptosis by downregulating GRP78 and CHOP protein levels. Conclusions: Abelmoschus manihot extract can be used as a promising therapeutic and preventive agent in the treatment of CIN.
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ObjectiveTo investigate the effect of Yishen Tongluo prescription （YSTLP） on apoptosis of renal tubular epithelial cells and explore the mechanism based on endoplasmic reticulum stress pathway of protein kinase R-like endoplasmic reticulum kinase （PERK）/activating transcription factor 4 （ATF4）/transcription factor C/EBP homologous protein （CHOP）. MethodThe db/db mice were randomly divided into model group， valsartan group （10 mg·kg-1）， and low， middle， high-dose YSTLP groups （1， 2.5， 5 g·kg-1）. Samples were collected after eight weeks of drug intervention. In addition， db/m mice in the same litter served as the control group. Human renal tubular epithelial cells （HK-2） were cultured in vitro and divided into the control group， advanced glycated end-product （AGE） group， and AGE + low， middle， and high-dose YSTLP groups （100， 200， 400 mg·L-1）. TdT-mediated dUTP nick end labeling （TUNEL） staining was used to detect the apoptosis rate of HK-2 cells. Methylthiazolyldiphenyl-tetrazolium bromide （MTT） assay was conducted to detect the viability of HK-2 cells. Calcium fluorescence probe staining and luciferase reporter gene method were adopted to detect the luciferase activity of folded protein response element （UPRE） and endoplasmic reticulum stress. Immunohistochemical （IHC） analysis was carried out to measure the protein expressions of phosphorylated PKR （p-PERK）， CHOP， and ATF4. Real-time polymerase chain reaction （Real-time PCR） was used to measure the mRNA expression levels of CHOP and X-box binding protein 1 （XBP1） in mouse kidney and HK-2 cells. Western blot was used to detect the protein expression level of p-PERK， PERK， CHOP， ATF4， B-cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， and cleaved Caspase-3 in mouse kidney and HK-2 cells. ResultIn the cellular assay， HK-2 cell viability was significantly reduced， and the apoptosis rate was elevated in the AGE group compared with the control group （P<0.01）. The mRNA and protein expression levels of apoptosis-related factor Bcl-2 were significantly reduced （P<0.01）， and those of Bax were significantly increased （P<0.01）. The protein expression level of cleaved Caspase-3 was significantly increased （P<0.01）. Compared with the AGE group， YSTLP administration treatment resulted in elevated cell viability and reduced apoptosis rate （P<0.01）. The mRNA and protein expression levels of Bcl-2 were significantly elevated in a time- and dose-dependent manner （P<0.01）， and those of Bax were significantly reduced in a time- and dose-dependent manner. The protein expression level of cleaved Caspase-3 was significantly reduced in a time- and dose-dependent manner （P<0.01）. The intracellular Ca2+ imbalance and UPRE luciferase fluorescence intensity were increased in the AGE group compared with the control group （P<0.01）. The mRNA levels of endoplasmic reticulum stress-related factors CHOP and XBP1 were significantly increased （P<0.01）， and the protein expression levels of p-PERK， CHOP， and ATF4 were significantly increased （P<0.05）. Compared with the AGE group， YSTLP effectively improved intracellular Ca2+ imbalance in HK-2 cells and decreased UPRE luciferase fluorescence intensity in a dose-dependent manner （P<0.01）. It reduced the mRNA levels of endoplasmic reticulum stress-related factors CHOP and XBP1 （P<0.01） and the protein expression levels of intracellular p-PERK， CHOP， and ATF4 in a dose- and time-dependent manner （P<0.01）. In animal experiments， the protein expression level of Bcl-2 was significantly reduced（P<0.01）， and that of cleaved Caspase-3 and Bax was significantly increased in the model group compared with the control group （P<0.05）. The protein expression level of Bcl-2 was dose-dependently elevated， and that of cleaved Caspase-3 and Bax was dose-dependently decreased in the YSTLP groups compared with the model group （P<0.01）. Compared with the control group， the mRNA expression levels of CHOP and XBP1 were significantly elevated in the model group （P<0.05， P<0.01）， and the protein expression levels of p-PERK， CHOP， and ATF4 were significantly increased （P<0.05）. Compared with the model group， YSTLP significantly decreased the mRNA expression levels of CHOP and XBP1 （P<0.01） and the protein expression levels of p-PERK， CHOP， and ATF4 （P<0.01）. ConclusionYSTLP can effectively inhibit endoplasmic reticulum stress and improve apoptosis of renal tubular epithelial cells， and its mechanism may be related to the regulation of the PERK/AFT4/CHOP pathway.
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A modified phaco chop technique for nuclear emulsification in posterior polar cataracts with nuclear sclerosis that avoids hydrodissection or nuclear rotation has been described. After performing a vertical chop maneuver to divide the nucleus, two pie-shaped nuclear fragments are removed from either side of the initial chop. The remaining nuclear fragments are then sequentially tumbled toward the center using the second instrument and emulsified while maintaining an intact epinuclear shell, which safeguards the fragile posterior capsule. The technique was successfully performed in 62 eyes of 54 patients with posterior polar cataract and grade II–IV nuclear sclerosis. The Chop and Tumble nucleotomy is a safe and effective technique for phacoemulsification in posterior polar cataracts with nuclear sclerosis, where hydrodissection and nuclear rotation are typically avoided
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Aims: the current research aimed to investigate LncRNA-MIAT in patients with nonHodgkin lymphoma (NHL) and to assess its correlation with clinicopathological features and treatment protocols of NHLs among Egyptian patients with Occult hepatitis C virus (HCV) infection (OCI). Patients & Methods: This study was conducted on 20 patients with NHL and 30 healthy subjects as the control group. All subjects were screened for HCV-RNA in both plasma and PBMCs. RT-PCR determined lncRNA-MIAT. Results: lncRNA-MIAT relative expression level was upregulated in NHL groups (2.73±0.86) compared to controls (1.06±0.07), P Ë0.001*. Among NHL, patients with OCI (3.2±0.63) had significantly higher levels of lncRNA-MIAT compared to HCV (2.6±1.08) and non-HCV (2.4±0.4), P Ë0.001*. Additionally, the relative expression levels of lncRNA-MIAT were significantly positively correlated with laboratory and clinicopathological features of NHL. Interestingly, concerning the treatment of DLBCLNHL, there were significantly higher levels of lncRNA-MIAT in no treatment subgroup (n=10, 3.31±0.95) compared to successfully treated subgroups [CHOP (n=7, 1.58±0.34) and R-CHOP (n=3, 11.16±0.21), P Ë0.001* Conclusions: lncRNA-MIAT level was upregulated in NHL patients, particularly patients with OCI. Thus, circulatory lncRNA-MIAT may serve as a promising non-invasive diagnostic marker for NHL associated with OCI
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Humans , Male , Female , Lymphoma, Non-Hodgkin , RNA, Long Noncoding , Myocardial Infarctionالملخص
Objective To explore the protective effect and mechanism of endoplasmic reticulum stress(ERS)signaling pathway PERK/eIF2α/ATF4/CHOP on BRL-3A cell injury by serum containing Zhige oral solution.Methods ①Preparation of drug-containing serum:SD male rats were randomly divided into 3 groups:Zhige oral solution group,Metadoxine group and normal group,and were gavaged with Zhige oral solution,Metadoxine and saline respectively,and the drug-containing serum was prepared by blood sampling from the abdominal aorta for 5 consecutive days of intervention.②Normal BRL-3A cells were cultured,and the cell survival rate of each group was determined by CCK-8 after 24h of cell intervention with different concentrations of ethanol(1.0%,1.5%,2.0%,2.5%,3.0%,5.0%,7.0%,10.0%).③Cultured normal BRL-3A cells were divided into normal group,model group,metadoxine group,and Zhige oral solution low,medium and high dose group(later referred to as low dose group,medium dose group and high dose group),After 24 h,the levels of γ-glutamyl transpeptidase(GGT)and lactate dehydrogenase(LDH)in the supernatant of each group were measured,and the survival rate of BRL-3A cells was measured by CCK-8.The expression levels of PERK/eIF2α/ATF4/CHOP pathway-related mRNA and protein in each group were measured by Real-time PCR and Western blot.Results ①After 24 h of cell intervention with the same concentration of ethanol(1.0%,1.5%,2.0%,2.5%,3.0%,5.0%,7.0%,10.0%),the cell survival rate gradually decreased with the increase of ethanol concentration,and when the ethanol concentration was 5%,the cell survival rate decreased significantly(P<0.05),so the ethanol concentrations of 1.0%,1.5%,2.0%,2.5%,3.0%,5.0%were chosen for the subsequent experiments.②Compared with the normal group,the GGT and LDH contents in the supernatant of the model group were significantly higher(P<0.05),and the expression of PERK/eIF2α/ATF4/CHOP pathway-related factors mRNA and protein were significantly higher(P<0.05),showing a significant state of liver injury.③Compared with the model group,the Zhige oral solution group and the metadoxine group showed different degrees of reduction in the above indexes,with the most significant effect in the Zhige oral solution high-dose group(P<0.05).Conclusion Serum containing Zhige oral solution ameliorated ethanol-induced hepatocyte injury in BRL-3A rats,and the mechanism may be related to the inhibition of endoplasmic reticulum stress PERK/eIF2α/ATF4/CHOP signaling pathway.
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AIM: To analyze the advantages of capsulorhexis-chop forceps assisted prechop(CCFP)technique in hard cataract phacoemulsification.METHODS: Prospective study. A total of 100 cases(100 eyes)with age-related grade IV hard cataract were included and randomly divided into 2 groups according to random number table, with 50 patients(50 eyes)in CCFP technique group(group A)and 50 cases(50 eyes)in stop-and-chop technique group(group B). The corneal endothelial cell count before and after operation, intraoperative US time, postoperative corneal endothelial cell loss rate, corneal edema grade at 1 and 7d and best corrected visual acuity(BCVA)were compared and statistically analyzed.RESULTS: The mean US time of group A was lower than that of group B [26.66(16, 40)s vs. 36.12(23, 46)s; Z=-5.56, P<0.01]. The mean corneal endothelial cell count in group A was higher than that in group B at 3mo after operation(2308.12±368.18cell/mm2 vs. 2104.06±379.87cell/mm2; t=2.728, P=0.008), and the loss rate of corneal endothelial cells in group A was lower than that in group B at 3mo after operation [10%(8%, 12%)vs. 17%(14%, 20%); Z=13.231, P<0.01]. The number of eyes with corneal edema of grade 0, 1, 2, 3 and 4 on 1d after surgery was 0, 23, 21, 6 and 0 in group A, respectively, while it was 0, 9, 26, 15 and 0 respectively in group B. Corneal edema in group A was less than that in group B(Z=10.514, P=0.005). The BCVA of group A was better than that of group B at 1d after operation, and there was significant difference in the number of eyes with different BCVA grades between the two groups(Z=7.176, P=0.029). There was no significant difference in the number of eyes with different grades of BCVA between the two groups at 3mo after surgery(Z=2.377, P=0.372).CONCLUSION: Compared with the stop-and-chop technique, CCFP technique uses less ultrasonic energy, has less damage to corneal endothelial cells and is suitable for hard cataract surgery.
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Renal tubular injury in patients with diabetic kidney disease(DKD) may be accompanied by glomerular and microvascular diseases. It plays a critical role in the progression of renal damage in DKD, and is now known as diabetic tubulopathy(DT). To explore the multi-targeted therapeutic effects and pharmacological mechanisms in vivo of total flavones of Abelmoschus manihot(TFA), an extract from traditional Chinese medicine for treating kidney disease, in attenuating DT, the authors randomly divided all rats into four groups: a normal control group(normal group), a DT model group(model group), a DT model+TFA-treated group(TFA group) and a DT model+rosiglitazone(ROS)-treated group(ROS group). The DT rat model was established based on the DKD rat model by means of integrated measures. After successful modeling, the rats in the four groups were continuously given double-distilled water, TFA suspension, and ROS suspension, respectively by gavage every day. After 6 weeks of treatment, all rats were sacrificed, and the samples of their urine, blood, and kidneys were collected. The effects of TFA and ROS on various indicators related to urine and blood biochemistry, renal tubular injury, renal tubular epithelial cell apoptosis and endoplasmic reticulum stress(ERS), as well as the activation of the protein kinase R-like endoplasmic reticulum kinase(PERK)-eukaryotic translation initiation factor 2α(eIF2α)-activating transcription factor 4(ATF4)-C/EBP homologous protein(CHOP) signaling pathway in the kidney of the DT model rats were investigated. The results indicated that hypertrophy of renal tubular epithelial cells, renal tubular hyperplasia and occlusion, as well as interstitial extracellular matrix and collagen deposition occurred in the DT model rats. Moreover, significant changes were found in the expression degree and the protein expression level of renal tubular injury markers. In addition, there was an abnormal increase in tubular urine proteins. After TFA or ROS treatment, urine protein, the characteristics of renal tubular injury, renal tubular epithelial cell apoptosis and ERS, as well as the activation of the PERK-eIF2α-ATF4-CHOP signaling pathway in the kidney of the DT model rats were improved to varying degrees. Therein, TFA was superior to ROS in affecting the pathological changes in renal tubule/interstitium. In short, with the DT model rats, this study demonstrated that TFA could attenuate DT by multiple targets through inhibiting renal tubular ERS-induced cell apoptosis in vivo, and its effect and mechanism were related to suppressing the activation of the PERK-eIF2α-ATF4-CHOP signaling pathway in the kidney. These findings provided preliminary pharmacological evidence for the application of TFA in the clinical treatment of DT.
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Rats , Animals , Abelmoschus , Reactive Oxygen Species/metabolism , Flavones/pharmacology , Endoplasmic Reticulum Stress , Diabetic Nephropathies/drug therapy , Apoptosis , Diabetes Mellitusالملخص
Purpose: To compare the clinical outcomes of endonucleation chop (EC) versus conventional crater (CC) chop techniques in phacoemulsification of hard nuclear cataracts. Methods: In this Prospective, longitudinal, randomized controlled study with double?blinding. Hundred consecutive eyes with uncomplicated cataracts (nucleus grades 3 and 4) were equally divided into Group A (EC) and Group B (CC). Intraoperative effective phacoemulsification time (EPT) and balanced salt solution (BSS) volume used was noted. Postoperative central corneal thickness (CCT), endothelial cell density (ECD), uncorrected Visual Acuity (UCVA), best?corrected visual acuity (BCVA), and IOP were recorded at days 1, 7, 30, and 90 postoperatively. The Chi?square test (categorical data), Mann–Whitney U test, and t?tests for other parameters were done. Results: The mean EPT in Groups A and B were 6.6 and 14.25 s, respectively (P < 0.001). The BSS volume used was 105.9 and 221.7 mL, respectively (P < 0.001). At 3 months, the ECD loss was 4.35 and 8.6%, respectively (P = 0.025). The?first day CCT was significantly increased in Group B but was the same in both groups at 3 months. A significant improvement in BCVA was noted in both groups. This new technique significantly reduces EPT, the BSS used, and ECD loss. Conclusion: Compared with the CC chop technique, the EC technique for phacoemulsification of hard nuclear cataracts conserved phacoemulsification energy and minimized exposure to the intraocular irrigating solution, provided a significant reduction in corneal endothelial damage, and led to faster visual rehabilitation
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Aim To investigate the effects of DAP on human rheumatoid arthritis synovial fibroblasts(RA-FLS)and its relationship with endoplasmic reticulum stress(ERS)PERK/ATF4/CHOP signaling pathway.Methods RA-FLS cells were cultured and identified by immunofluorescence assay for Vimentin and CD68.CCK-8 detected(0,5,10,20,30,40,50,60,70)mg·L-1 DAP on the proliferation activity of RA-FLS cells.According to the proliferation activity,the experiment was divided into blank group(Blank),low dose group(L-DAP),medium dose group(M-DAP),high dose group(H-DAP)and high dose+specific inhibitor 4-PBA group(H-DAP+4-PBA).The levels of TNF-α and IL-6 were detected by ELISA.Cell apoptosis was detected by flow cytometry.The invasion and migration of cells were detected by Transwell and scratches assays,and the expressions of endoplasmic reticulum stress-related proteins GRP78,PERK,P-PERK,ATF4,CHOP,Caspase-12,C-Caspase-12 and Bcl-2 were assessed by Western blot.Results CCK-8 results showed that compared with 0 mg·L-1 DAP group,the proliferation activity of each group in 20-70 mg·L-1 DAP group was significantly different(P<0.05),and the proliferation rate corresponding to 0,20,40 and 60 mg·L-1 DAP group was significantly different(P<0.05).This concentration was used as the basis for blank,low-dose,medium-dose,high-dose groups and high-dose+4-PBA experimental grouping.DAP could inhibit the release of inflammatory cytokines IL-6 and TNF-α from RA-FLS cells in concentration,reduce the invasion ability of cells,promote apoptosis,upregulate the expression of PERK,P-PERK,ATF4,GRP78,CHOP,Caspase-12, C-caspase-12 proteins and decrease the expression level of anti-apoptotic protein Bcl-2.Another set of experiments demonstrated that high dose DAP+4-PBA could up-regulate the expression of IL-6 and TNF-α,as well as the invasion of cells,and inhibit the expression of apoptosis and ERs-related proteins.Conclusions Daphresin regulates the secretion of inflammatory factors by activating the PERK/ATF4/CHOP signaling pathway,inhibits the proliferation and invasion of RA-FLS cells,and induces apoptosis,which is expected to be a potential therapeutic pathway for RA.
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Objetivo: Describir la posible presencia de complicaciones posoperatorias en pacientes diabéticos tipo 2, operados de catarata por la técnica de facochop. Métodos: Se realizó un estudio observacional, descriptivo, y longitudinal prospectivo, con 128 pacientes diabéticos (168 ojos) operados de catarata en el Centro de Microcirugía Ocular del Instituto Cubano de Oftalmología "Ramón Pando Ferrer", entre septiembre del año 2017 y diciembre de 2018. Las variables clínicas y epidemiológicas objeto del estudio fueron la edad, el sexo, el color de la piel, la queratometría, la profundidad de la cámara, la longitud axial, la dureza del cristalino, la presión intraocular pre- y posoperatoria y las complicaciones posoperatorias en las primeras 72 horas. Resultados: En el estudio predominaron las mujeres (63,3 por ciento), de piel blanca (74,2 por ciento), quienes presentaron ojos biométricamente normales, con una diferencia estadísticamente significativa. La presión intraocular aumentó a las 24 horas de la cirugía. De los 128 pacientes diabéticos tipo 2 operados de catarata senil por la técnica de facochop, se pudo determinar que el promedio y la desviación estándar de la edad se ubicaron en los 69,49 ± 8,96 años, y osciló entre los 50 y 88 años. Conclusiones: Las complicaciones posoperatorias más frecuentes son el edema corneal y la uveítis. Es significativamente más probable encontrar la uveítis posquirúrgica en pacientes menores de 70 años, y el edema corneal en quienes sobrepasan esta edad(AU)
Objective: Describe the possible presence of postoperative complications in type 2 diabetic patients undergoing phaco chop cataract surgery. Methods: An observational prospective longitudinal descriptive study was conducted of 128 diabetic patients (168 eyes) undergoing cataract surgery at the Center for Ocular Microsurgery of Ramón Pando Ferrer Cuban Institute of Ophthalmology from September 2017 to December 2018. The clinical and epidemiological variables analyzed were age, sex, skin color, keratometry, chamber depth, axial length, crystalline hardness, pre- and postoperative intraocular pressure, and postoperative complications in the first 72 hours. Results: A predominance was found of women (63.3 percent) of white skin color (74.2 percent) and biometrically normal eyes, with a statistically significant difference. Intraocular pressure rose 24 hours after surgery. Mean age and standard deviation were 69.49 ± 8.96 years, range of 50-88 years, in the 128 type 2 diabetic patients undergoing senile cataract surgery by the phaco chop technique. Conclusions: The most common postoperative complications are corneal edema and uveitis. It is significantly more probable to find postsurgical uveitis in patients aged under 70 years and corneal edema in patients aged over 70 years(AU)
الموضوعات
Humans , Female , Adult , Middle Aged , Aged , Postoperative Complications/therapy , Uveitis/complications , Cataract Extraction/methods , Diabetes Mellitus, Type 2/etiology , Microsurgery/methods , Corneal Edema/complications , Epidemiology, Descriptive , Prospective Studies , Longitudinal Studies , Observational Studies as Topicالملخص
Objective:To investigate the effect of Ziziphi Spinosae Semen （ZSS） and Albiziae Flos （AF） on behavior and endoplasmic reticulum stress endoplasmic reticulum stress protein kinase R-like endoplasmic reticulum kinase （PERK）/activated transcription factor 4 （ATF4）/CCAAT enhancer binding protein （CHOP） pathway in depression model rats， and to explore its antidepressant mechanism. Method:The male SD rats were divided into normal group， model group， ZSS-AF high dose， middle dose and low dose groups （16， 8， 4 g·kg<sup>-1</sup>） and Venlafaxine group （0.008 g·kg<sup>-1</sup>）， <italic>n</italic>=15 in each group. Except the normal group， the depression model was established in the rats of other 5 groups by the method of chronic unpredictable mild stress （CUMS） combined with isolated feeding. The normal group and model group were given with distilled water by gavage when modeling， while other groups received corresponding drug by intragastric administration for 21 days. Behavior changes of rats in each group were observed by the open field test and sugar water consumption test on 1<sup>th </sup>and 21<sup>th</sup>day of the experiment. The protein expressions of PERK， CHOP， B-cell lymphoma-2 associated X protein （Bax） and cysteine-containing aspartate-specific proteases-3（Caspase-3） were detected by Western blot（WB）， the ultrastructural changes of the hippocampus were observed by transmission electron microscope， the apoptosis of hippocampal neurons was observed by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL） method. Result:Compared with the normal group， the scores of open field test and sugar water consumption rate in model group rats decreased （<italic>P</italic><0.01）. Compared with the model group， the scores of open field test and water consumption rate increased （<italic>P</italic><0.01） in ZSS-AF groups and Venlafaxine group. Transmission electron microscope showed that the changes of neuronal damage in hippocampal were revealed in the model group， whereas those neuronal damages were relieved in ZSS-AF groups and Venlafaxine group. TUNEL method showed that the number of apoptotic neurons in hippocampal increased in the model group （<italic>P</italic><0.01）， but decreased in ZSS-AFgroups and Venlafaxine group （<italic>P</italic><0.01）. WB results showed that as compared with the normal group， protein expressions of PERK， CHOP， Bax and Caspase-3 were up-regulated significantly in the model group （<italic>P</italic><0.01）， whereas those were down-regulated in ZSS-AF groups and Venlafaxine group （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:The antidepressant effect of ZSS-AF herbal pair may be correlated with the regulation of endoplasmic reticulum stress PERK/ATF4/-CHOP pathway.
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@#AIM: To analyze the efficacy of bevel-up phaco chop versus bevel-down phaco drill in the treatment of hard nuclear cataract and the influence on corneal endothelium.<p>METHODS: Between August 2018 and April 2020, 94 patients(104 eyes)with hard(grade Ⅳ-Ⅴ)nuclear cataract treated with phacoemulsification were enrolled in this retrospective study. Among them, 50 eyes treated by bevel-up phaco chop were included in the control group, while 54 eyes treated by phaco drill technique were included in the observation group. The time of using ultrasound, ultrasonic energy and accumulated energy complex parameter in the two groups were recorded. The best corrected visual acuity(BCVA)in 7d, corneal endothelial cell count in 2mo, the proportions of normal hexagonal cells and the degree of corneal edema in 7d were compared between the two groups.<p>RESULTS: The time of using ultrasound, ultrasonic energy and accumulated energy complex parameter of the observation group were shorter and lower than those of the control group(<i>P</i><0.05). The BCVA of both groups increased after operation, and the observation group had higher BCVA than the control group at the same time(<i>P</i><0.05). In 1 and 2mo, the corneal endothelial cell counts and the proportions of normal hexagonal cells in both groups were lower than those before operation. Meanwhile, the corneal endothelial cell count and the proportions of normal hexagonal cells in the observation group were higher than those in the control group(<i>P</i><0.05). Besides, corneal edema was milder in the observation group than in the control group at 24h and 7d after operation(<i>P</i><0.05).<p>CONCLUSION: Compared with bevel-up phaco chop, phaco drill can significantly shorten the time of using ultrasound, reduce ultrasound energy, promote visual recovery, and reduce corneal endothelial cell damage.
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To explore the protective effect and mechanism of ethyl acetate extract from Bidens bipinnata on hepatocyte damage induced by endoplasmic reticulum stress. Tunicamycin was used to establish the damage model in L02 cells. Methyl thiazolyl tetrazolium(MTT) colorimetric assay was used to investigate the survival rate of ethyl acetate extract from B. bipinnata in L02 cells injury induced by endoplasmic reticulum stress; the protein expressions of endoplasmic reticulum stress-related molecule glucose regulated protein 78(GRP78), PKR-like ER kinase(PERK), eukaryotic initiation factor-2(eIF2α), activating transcription factor 4(ATF4), C/EBP homologous protein(CHOP), B-cell CLL/lymphoma 2(Bcl-2), Bal-2 associated X apoptosis regulator(Bax) were examined by Wes-tern blot. The expressions of the above proteins were also detected after endoplasmic reticulum stress inhibitor(4-phenyl butyric acid) and CHOP shRNA-mediated knockdowns were added. The expressions of GRP78, PERK, CHOP in L02 cells were observed by immunofluorescence method. The results showed that ethyl acetate extract from B. bipinnata could significantly increase the survival rate of L02 cell injury caused by endoplasmic reticulum stress in a dose and time-dependent manner(P<0.05 or P<0.01). The expression levels of GRP78, PERK, eIF2α, ATF4, CHOP and Bax in the drug treatment groups were significantly down-regulated(P<0.05 or P<0.01), while Bcl-2 was significantly up-regulated(P<0.01). After endoplasmic reticulum stress inhibitor and CHOP shRNA-mediated knockdowns were added, the expression levels of GRP78, PERK, eIF2α, ATF4, CHOP, Bax in the drug treatment groups were significantly down-regulated(P<0.01), whereas Bcl-2 was significantly up-regulated(P<0.01). Immunofluorescence results showed that the expressions of GRP78, PERK, CHOP were consistent with the Western blot method. In conclusion, ethyl acetate extract from B. bipinnata has a significant protective effect on the damage of L02 cells caused by endoplasmic reticulum stress. The mechanism may be related to the inhibition of endoplasmic reticulum stress and the down-regulation of apoptosis in cells through the PERK/eIF2α/ATF4/CHOP signaling pathway.
الموضوعات
Acetates , Apoptosis , Bidens , Endoplasmic Reticulum Stress , Hepatocytes , Transcription Factor CHOP/genetics , eIF-2 Kinase/geneticsالملخص
@#AIM: To discuss the advantages of press-and-chop technique compared with Nagahara phaco-chop technique.<p>METHORDS: Totally 70 patients(70 eyes)with age-related cataract were randomly divided into 2 groups, press-and-chop technique group(35 patients 35 eyes), phaco-chop technique group(35 patients 35 eyes). In all cases, surgery began with a clear corneal incision, capsulorhexis and hydrodissection. In the press-and-chop technique group, the superficial cortex and epinucleus were aspirated by the phaco tip, then press the center of the lens front surface with the phaco tip. The Nagahara chopper was set around the lens equator, then the phaco tip was driven into the nucleus from the main incision, pull the Nagahara chopper toward the phaco tip. The two instruments were then separated laterally to produce a complete fracture of the nucleus. In phaco-chop technique group, the phaco tip was buried in the center of the endonucleus. The Nagahara chopper was brought through the side-port incision and the equator of endonucleus was engaged by the chopper under the lower edge of the capsulorhexis and pulled toward the phaco tip. The 2 instruments were then separated laterally to produce a complete fracture of the nucleus. The U/S time, preoperative and postoperative corneal endothelial cell density, corneal endothelium loss rate, corneal edema at 1d, 7d, best corrected visual acuity before and after surgery were recorded.<p>RESULTS: The U/S time of press-and-chop technique group was lower than phaco-chop technique group [12.76(8.76,16.76)s <i>vs</i> 22.87(18.36, 27.38)s, <i>P</i><0.01]. The corneal endothelial cells density in press and chop technique group was higher than that in phaco-chop technique group 1mo after operation(2133.44±348.58/mm<sup>2</sup> <i>vs</i> 1957.94±280.54/mm<sup>2</sup>, <i>P</i><0.05), and the variation rate of corneal endothelial cells in press-and-chop technique group was lower than that in phaco-chop technique group 1mo after surgery [0.15(0.08,0.22)<i>vs</i> 0.22(0.16, 0.28), <i>P</i><0.01]. The corneal edema in press-and-chop technique group was lighter than that in phaco-chop technique group on the first day after surgery(<i>Z</i>=13.195, <i>P</i>=0.004), and corneal edema in both groups subsided on the 7d after surgery. There was no significant difference between two groups in BCVA on the first day after surgery(<i>Z</i>=-0.48, <i>P</i>=0.63).<p>CONCLUSION: Compared with Nagahara phaco-chop technique, press-and-chop technique is simple and safe with less complications.
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Aim To observe the protective effect of Gancao Xiexin Decoction (GXD) on intestinal epithelial cells (IECs) under endoplasmic reticulum stress from the perspective of PERK-elF2α-CH0P signaling pathway. Methods Caco-2 cells were cultured and divided into normal control group (NC), model control group(MC), low-dose GXD group(GXD-L), mediumdose GXD group (GXD-M) and high-dose GXD group (GXD-H). Models of the stress epithelial cells were induced by tunicamycin(Tm), and GXD groups were treated with GXD contained serum at the same time. The cell survival rate was assessed by CCK-8 method, the cell apoptotic rate and cell cycle distribution were determined by flow cytometry, and the cell barrier permeability was determined by TEER and FITC-dextran method; the expression levels of core proteins of PERK-elF2α-CH0P signaling pathway were detected by Western blot. Results Compared with MC group, GXD intervention could improve cell survival rate(P < 0. 05), reduce their apoptotic rate(P <0. 01), relieve cell cycle arrest(P < 0. 01), improve cell barrier permeability by increasing cell TEER value (P < 0. 01) and decreasing FITC-dextran concentration (P < 0.05), and the levels of p-PERK, p-elF2α, ATF4 and CHOP in GXD-M and GXD-H groups all visibly descended (P < 0. 01). Conclusions GXD can reduce the excessive apoptosis of IECs and protect intestinal epithelial cell barrier homeostasis by inhibiting the signal transduction of PERK-elF2α-CH0P apoptotic pathway.