Mechanism of Yishen Tongluo Prescription in Inhibiting Endoplasmic Reticulum Stress and Improving Apoptosis of Renal Tubular Epithelial Cells Based on PERK/ATF4/CHOP / 中国实验方剂学杂志

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.

Chop and tumble nucleotomy – A technique for safe nuclear emulsification in posterior polar cataract

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

Effects and mechanisms of total flavones of Abelmoschus manihot in attenuating diabetic tubulopathy by targeting endoplasmic reticulum stress-induced cell apoptosis / 中国中药杂志

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.

Clinical significance of LncRNA-MIAT as a non-invasive diagnostic marker of nonHodgkin's lymphoma associated with occult hepatitis C virus infection

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 ˂0.001*. 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 ˂0.001*. 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 ˂0.001* 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

Application of capsulorhexis-chop forceps assisted prechop technique in hard cataract phacoemulsification / 国际眼科杂志(Guoji Yanke Zazhi)

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&#x003C;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&#x003C;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.

Comparative evaluation of modified crater (endonucleation) chop and conventional crater chop techniques during phacoemulsification of hard nuclear cataracts: A randomized study

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

Complicaciones posoperatorias inmediatas en pacientes diabéticos tipo 2 operados de catarata por la técnica de facochop / Immediate postoperative complications in type 2 diabetic patients undergoing phaco chop cataract surgery

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)

Protective effect of ethyl acetate extract from Bidens bipinnata on hepatocyte damage induced by endoplasmic reticulum stress / 中国中药杂志

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.

Application of different phacoemulsification and chopping techniques in hard nuclear cataract surgery / 国际眼科杂志(Guoji Yanke Zazhi)

@#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.

Comparison of press-and-chop and Nagahara phaco-chop phacoemulsification nucleotomy techniques / 国际眼科杂志(Guoji Yanke Zazhi)

@#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/mm² <i>vs</i> 1957.94±280.54/mm², <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.

Effects of Ziziphi Spinosae Semen and Albiziae Flos on Behavior and Endoplasmic Reticulum Stress PERK/ATF4/CHOP Pathway in Depression Model Rats / 中国实验方剂学杂志

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^-1） and Venlafaxine group （0.008 g·kg^-1）， <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^thand 21^thday 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.

Comparing the number of attempts required for complete nuclear chop using calibrated and conventional phacotip

Background: Phacoemulsification is a state of art technique with a steep learning curve the configuration of the phacotip affects the efficacy and execution of the nuclear chopping techniques. Inadequate penetration of phacotip may result in partial thickness nuclear cleavage with residual posterior plane and over enthusiastic penetration may result in posterior capsule rupture. This may be avoided if some estimate can be made preoperatively of the depth of penetration required to achieve full thickness crack.Methods: A total of 60 eyes of 60 patients with age related cataract with grade 4.0 to 6.9 (LOCS III) fulfilling the inclusion and exclusion criteria were included in the study after written informed consent. They were divided into two groups A and B with 30 patients each randomly using envelope method and were operated using conventional and calibrated phacotips respectively and the number of attempts required to achieve complete nuclear chop was noted.Results: In group A, vertical chop was safely and effectively done is 23 patients in just one attempt while it took 2 attempts in 6 patients and 3 attempts in 1 patient. In group B, it took just one attempt in 26 patients while 2 attempts were required in 4 patients. The two groups were comparable (p value 0.453) and was found to be statistically significant.Conclusions: With the use of a calibrated phacotip, the surgeon already knows to what depth he has to penetrate the tip into the center of nucleus depending upon the grade of cataract which results in safe and effective chop in minimum attempts where as in the conventional phacotip, it is more of a blind process. Hence the calibrated phacotip has taken the guess work out of question.

Saikosaponin-d simultaneously induces apoptotic and paraptosis-like cell death in hepatoma cells / 西安交通大学学报(医学版)

Objective: To investigate the cell death-inducing effect of saikosaponin-d (SSD) on human hepatoma Hep3B cells and its potential mechanism. Methods: Hepatoma Hep3B cells were divided into five groups: blank control group, DMSO (vehicle) group, and three SSD treatment groups treated with various doses of SSD (5, 10, and 15 μmol/L). MTT assay was used to evaluate cell viability. Flow cytometer was employed to quantitatively detect the percentage of dead cells after Annexin V-FITC/PI double staining. Apoptosis was detected morphologically after Hoechst 33258 staining. The activity of caspase-3 apoptotic protease was determined by spectrophotometry. Cell morphologic changes were observed with an inverted microscope. Western blotting and Real-time PCR were employed to evaluate the expression levels of C/EBP homology (CHOP) protein and mRNA, respectively. Results: MTT assay showed that SSD inhibited the viability of human hepatoma Hep3B cells in concentration- and time-dependent manners. Sinomenine hydrochloride induced the death of Hep3B cells in a concentration-dependent manner indicated by flow cytometry. After staining with Hoechst 33258, the nuclei of SSD-treated cells showed nucleosomal agglutination, nucleosomal shrinkage and fragmentation under the fluorescence microscope, which are the characteristics of apoptotic cells. SSD significantly activated the key apoptotic executor caspase-3. The occurrence of paraptosis, characterized by extensive cytoplasmic vacuoles, was observed in SSD-treated cells under an inverted microscope. The pretreatment of a pancaspase inhibitor Z-VAD-FMK completely inhibited caspase-3 activity triggered by SSD, but only partially suppressed cell death and could not reduce the cytoplasmic vacuolation in SSD-treated cells. The protein and mRNA expressions of CHOP, a stress-inducible molecule, were upregulated by SSD, which could not be inhibited by Z-VAD-FMK. Conclusion: SSD can simultaneously induce caspase-dependent apoptosis and caspase-independent paraptosis in human hepatoma Hep3B cells. The upregulated expression of CHOP may be the mechanism involved in SSD-induced paraptosis.

Retrospective study on outcomes of terminal chop technique of nuclear fragmentation in phacoemulsification and combined surgery: An observational study

Purpose: Terminal chop, a new technique of nuclear segmentation, has been recently described in literature. The technique is considered to be a safe and effective option. Methods: We did a retrospective study to evaluate the outcome of terminal chop. Patients operated for terminal chop (phacoemulsification with or without trabeculectomy) from October 2016 to July 2018 were included in the study. Results: In all, 933 patients were included in the study. The grade of nuclear sclerosis ranged from 2 to 5. All the patients had a significant visual improvement at 1 month postop. Around 3.4% had pseudoexfoliation and 8.89% of the eyes underwent combined surgery. Cumulative dissipated energy showed a proportionate relation with the grade of nucleus. Intraoperative complication rate was 0.08%. Postoperative complication rate was 3.50%. Conclusion: Terminal chop proved to be a safe, reproducible, and effective technique of nuclear segmentation in different grades of cataract.

Effects of difluoromethylalanine on cardiac hypertrophy and endoplasmic reticulum stress in type 2 diabetic rats / 中国药理学通报

Aim: To investigate the protective effects of difluoromethylornithine (DFMO) on cardiac hypertrophy and to explore its influence on the protein expression of GRP78 and CHOP in type 2 diabetic rats. Methods: The type 2 diabetic mellitus (T2DM) rat model was induced by 4 weeks' high sucrose-high fat diet feeding and one-time intraperitoneal injection of streptozotocin. Rats were randomly divided into three groups:control group, T2DM model group and DFMO treatment group. Rats of treatment group were treated with 2% DFMO drinking for 12 weeks. The changes in blood glucose and cardiac index were calculated. Myocardium superoxide dismutase (SOD), malondialdehyde (MDA) and the total antioxidant capacity (T- A0C) were determined. The morphological changes were observed by Masson staining, and the protein expressions of GRP78 and CHOP were measured by Western blot. Results: Compared with T2DM model group, the blood glucose, cardiac index and the degree of myocardial fibrosis significantly decreased (P < 0. 05), the content of MDA decreased and the activity of SOD and T-AOC increased. Meanwhile, the protein expressions of GRP78 and CHOP were down-regulated in treatment group (P < 0. 05). Conclusions: DFMO exerts protective effects on cardiac hypertrophy in T2DM rats, which may be related to its function of the down-regulation of protein expression of GRP78 and CHOP and inhibition of endoplasmic reticulum stress.

Efficacy analysis of rituximab combined with CHOP or EPOCH regimen in treatment of diffuse large B-cell lymphoma with newly diagnosed myc/bcl-2 double-expression / 白血病·淋巴瘤

Objective To evaluate the efficacy, safety and the prognosis of rituximab combined with CHOP (R-CHOP) or combined with EPOCH (R-EPOCH) regimen in treatment of newly diagnosed myc/bcl-2 double-expression diffuse large B-cell lymphoma (DLBCL). Methods The clinical efficacy, adverse reactions and survival of 16 DLBCL patients who received R-CHOP regimen with double-expression and 15 DLBCL patients who received R-EPOCH regimen with double-expression between August 2014 and December 2017 in the First Affiliated Hospital of Zhengzhou University were retrospectively analyzed, and all patients with double or triple-hit lymphoma were excluded. Results The ratio of the International Prognostic Index (IPI) score ≥3 in R-EPOCH group was higher than that in R-CHOP group [10 cases vs. 5 cases, χ2＝ 3.888, P＝0.049]. There were no statistical differences in other clinical data of both groups. Complete remission (CR) was achieved in 75% (12/16) of R-CHOP group and 67% (10/15) of R-EPOCH group, and there was no statistical difference between the two groups (χ2＝ 0.013, P＝ 0.908). The incidence of grade 3-4 leukopenia and grade 3-4 thrombocytopenia in R-EPOCH group was higher than that in R-CHOP group, and the difference was statistically significant [93% (14/15) vs. 38% (6/16), χ 2＝ 10.542, P＝ 0.01; 73% (11/15) vs. 6% (1/16), χ 2＝ 14.685, P< 0.01]. The 1-year overall survival (OS) rate and progression-free survival (PFS) rate was 86.5% and 81.3% in R-CHOP group, 76.0% and 51.4% in R-EPOCH group. There was no statistical difference in the 1-year OS rate between the two groups (P＞0.05), and the 1-year PFS rate in R-CHOP group was higher than that in R-EPOCH group (P<0.01). The 1-year OS rate and PFS rate was 100.0% and 93.8% in IPI<3 group, 65.5% and 45.0% in IPI≥3 group. The 1-year OS rate and PFS rate in IPI<3 group were superior to those in IPI≥3 group (all P ＞0.01). The 1-year OS rate and PFS rate of the female group were lower than those of the male group (59.6% vs. 100.0%, 44.2% vs. 86.3%), and there were statistical differences (all P<0.01). Cox regression analysis showed that high IPI score was an independent prognostic factor for all patients ( HR＝6.335, 95% CI 0.740-54.261, P＝0.092). Conclusions R-EPOCH and R-CHOP are the ideal treatment methods for the double-expression lymphoma. Both regimens have similarities in the treatment outcome of myc/bcl-2 double-expression DLBCL. The adverse reactions in R-EPOCH group are severer than those in R-CHOP group.

Endoplasmic Reticulum Stress Induces MUC5AC and MUC5B Expression in Human Nasal Airway Epithelial Cells

OBJECTIVES: Endoplasmic reticulum (ER) stress is known to be associated with inflammatory airway diseases, and three major transmembrane receptors: double-stranded RNA-activated protein kinase-like ER kinase, inositol requiring enzyme 1, and activating transcription factor 6 (ATF6) play important roles in ER stress-related proinflammatory signaling. However, the effects of ER stress and these three major signaling pathways on the regulation of the production of airway mucins in human nasal airway epithelial cells have not been elucidated. METHODS: In primary human nasal epithelial cells, the effect of tunicamycin (an ER stress inducer) and 4-phenylbutyric acid (4-PBA, ER stress inhibitor) on the expression of MUC5AC and MUC5B was investigated by reverse transcriptasepolymerase chain reaction, real-time polymerase chain reaction, enzyme immunoassay, and immunoblot analysis. Small interfering RNA (siRNA) transfection was used to identify the mechanisms involved. RESULTS: Tunicamycin increased the expressions of MUC5AC and MUC5B and the mRNA expressions of ER stress-related signaling molecules, including spliced X-box binding protein 1 (XBP-1), transcription factor CCAAT-enhancer-binding protein homologous protein (CHOP), and ATF6. In addition, 4-PBA attenuated the tunicamycin-induced expressions of MUC5AC and MUC5B and the mRNA expressions of ER stress-related signaling molecules. Furthermore, siRNA knockdowns of XBP-1, CHOP, and ATF6 blocked the tunicamycin-induced mRNA expressions and glycoprotein productions of MUC5AC and MUC5B. CONCLUSION.: These results demonstrate that ER stress plays an important role in the regulation of MUC5AC and MUC5B via the activations of XBP-1, CHOP, and ATF6 in human nasal airway epithelial cells.

Dose-Dense Rituximab-CHOP versus Standard Rituximab-CHOP in Newly Diagnosed Chinese Patients with Diffuse Large B-Cell Lymphoma: A Randomized, Multicenter, Open-Label Phase 3 Trial / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Rituximab with cyclophosphamide, doxorubicin, vincristine, and prednisone administered every 3 weeks (R-CHOP-21) is the standard care for diffuse large B-cell lymphoma (DLBCL). It is unknown whether the dose-dense R-CHOP (R-CHOP-14) could improve the outcome of the disease in Asian population. MATERIALS AND METHODS: Newly diagnosed DLBCL patients were centrally, randomly assigned (1:1) to receive R-CHOP-14 or R-CHOP-21. R-CHOP-14 was administered every 2 weeks, and R-CHOP-21 was administered every 3 weeks. Primary end point was disease-free survival (DFS). Secondary end points included overall survival (OS), progression-free survival (PFS), response rate and toxicities. RESULTS: Seven hundred and two patients were randomly assigned to receive R-CHOP-14 (n=349) or R-CHOP-21 (n=353). With a median follow-up of 45.6 months, the two groups did not differ significantly in 3-year DFS (79.6% for R-CHOP-14 vs. 83.2% for R-CHOP-21, p=0.311), 3-year OS (77.5% for R-CHOP-14 vs. 77.6% for R-CHOP-21, p=0.903), or 3-year PFS (63.2% for R-CHOP-14 vs. 66.1% for R-CHOP-21, p=0.447). Patients with an International Prognostic Index (IPI) score ≥ 2 had a poorer prognosis compared to those with an IPI score < 2. Grade 3/4 hematologic and non-hematologic toxicities were manageable and similar between R-CHOP-14 and R-CHOP-21. CONCLUSION: R-CHOP-14 did not improve the outcome of DLBCL compared to R-CHOP-21 in Asian population. With manageable and similar toxicities, both of the two regimens were suitable for Asian DLBCL patients. For high-risk patients with IPI ≥ 2, new combination regimens based on R-CHOP deserve further investigation to improve efficacy.

Effects of Salvianolic acid on endoplasmic reticulum stress pathway in brain hippocampus of PAP mice / 中国中西医结合急救杂志

Objective To investigate the effect of Salvianolic acid on endoplasmic reticulum stress (ERS) pathway in brain hippocampus of PAP mice. Methods Twenty PAP dual transgenic male mice were selected, they were randomly divided into a PAP mice model group and a Salvianolic acid group, 10 mice in each group; another 10 SPF grade C57BL/6J male mice were selected as a normal control group. In the Salvianolic acid group, 0.9% normal saline solution of Salvianolic lyophilized injection (400 g/L) of dosage 21 mg·kg-1·d-1 was injected intravenously through a tail vein of mice; the PAP mice model and normal control groups were given the same amount of 0.9% normal saline, and the therapeutic course was consecutive 4 weeks in the three groups. At the end of the 4th week, the Morris water maze test was carried out to observe the changes of escape latency, the third quadrant residence time (RTQ), entry angle into water and cross-platform times of mice in each group; amyloid precursor protein (APP) positive cell expression in cerebral hippocampus of mice were detected by immunohistochemistry; Western Blot was used to detect the expression level of PER like endoplasmic reticulum kinase-eukaryon initiation factor 2α-C/EBP homogenous protein (PERK-eIF2α-CHOP) pathway related proteins in hippocampus of mice. Results The escape latency of the PAP mice model group on the 1st to 5th day were significantly longer than those of the normal control group, although a downward trend was observed on the 5th day, it was still significantly longer than that of the model group (seconds: 58.41±2.36 vs. 28.60±10.15); compared with the PAP mice model group, the escape latency of Salvianolic acid group was shorter at each time point, and reached the shortest level on the 5th day (seconds: 31.97±8.36 vs. 58.41±2.36). In the PAP mice model group, the RTQ and the number of crossing platforms were significantly lower than those in the normal control group [RTQ (seconds): 8.27±2.95 vs. 15.97±7.33, numbers of crossing platforms (frequency/90 s): 0.70±0.95 vs. 2.70±0.48]; the entry angle was obviously greater than that of the normal control group [(47.94±4.68)°vs. (32.66±2.55)°, P < 0.05]. Compared with PAP mice model group, in Salvianolic acid group, the RTQ and number of crossing platform were significantly higher [RTQ (seconds): 13.57±1.86 vs. 8.27±2.95, number of crossing platforms (frequency/90 s):1.60±0.97 vs. 0.70±0.47], the entry angle was markedly smaller [(35.46±6.79)°vs. (47.94±4.68)°,P < 0.05]. The positive expression rate of APP and the protein expressions of CHOP, p-eIF2α in PAP mice model group were significantly higher than those in the normal control group [the positive rate of APP: (60.44±6.19)% vs. (21.05±5.87)%, CHOP protein expression (gray value): 3.09±0.07 vs. 1.46±0.09, p-eIF2αprotein expression (gray value): 0.98±0.09 vs. 0.47±0.06, all P < 0.01], the expression of PERK and p-PERK were lower than those in normal control group [PERK (gray value): 0.42±0.06 vs. 0.82±0.11, p-PERK protein expression (gray value): 0.98±0.09 vs. 0.64±0.10, both P < 0.01]; the positive expression rate of APP and protein expressions of CHOP, p-eIF2α in Salvianolic acid group were significantly lower than those in PAP mice model group [positive expression rate of APP: (33.09±10.33)% vs. (60.44±6.19)%, CHOP protein expression (gray value): 1.57±0.12 vs. 3.09±0.07, p-eIF2α protein expression (gray value): 0.80±0.07 vs. 0.98±0.09, all P < 0.01], while PERK and p-PERK expression were significantly higher than those in the model group [PERK (gray value): 0.89±0.12 vs. 0.42±0.06, p-PERK (gray value): 0.78±0.08 vs. 0.98±0.09, both P < 0.01]. Conclusion Salvianolic acid might work through the PERK-eIF2α-CHOP pathway to reduce the retention of APP in the hippocampus tissue of PAP dual-transgenic mice, thereby the learning ability of the mice is improved, and the progression of brain injury delayed.

Linfoma de la zona gris con características intermedias entre un linfoma difuso de células B grandes y un linfoma Hodgkin clásico. Reporte de un caso y revisión de la literatura / Grey zone lymphoma with features intermediate between diffuse large B-cell lymphoma and classical Hodgkin lymphoma. Case report and literature review

Resumen Entre los linfomas de la zona gris (LZG) encontramos neoplasias con características compartidas entre un linfoma difuso de células B grandes (LDCBG) y un linfoma de Hodgkin clásico (LHC). Lo poco habitual de la patología combinado con la heterogenicidad de la enfermedad, su reciente descripción como entidad específica que conlleva a dificultad y reto diagnóstico, así como la falta de suficiente experiencia terapéutica hacen de la enfermedad una entidad compleja de difícil diagnóstico y reto terapéutico que justifica su descripción continua. Se presenta una paciente con fiebre de un mes sin respuesta al manejo inicial, se estudió y realizó biopsia de ganglio inguinal izquierdo con resultado diagnóstico de LZG con características intermedias entre LDCBG y LHC. Aunque no existen guías establecidas para el manejo de esta entidad, la evidencia actual sugiere mejor respuesta en tratamientos dirigidos a LDCBG, misma terapia empleada en esta paciente con la cual se obtuvo respuesta favorable.

Abstract In the grey zone lymphomas (GZL), there are overlapping characteristics between diffuse large B-cell lymphoma (DLBCL) and classic Hodgkin lymphoma (CHL). The unusual nature of the pathology combined with the heterogeneity of the disease, its recent description as a specific entity, its diagnostic difficulty, and the lack of sufficient therapeutic experience justifies its continuous description. The case is presented of a patient with a fever of one month onset, with no response to initial management. A left inguinal lymph node biopsy reported a diagnosis of GZL with intermediate characteristics between DLBCL and CHL. Although there are no established guidelines for the management of this condition, the current evidence suggests a better response in treatments meant for diffuse large B-cell lymphoma. This same therapy was used in this patient, with a favourable clinical outcome.