1.Total saponins from Trillium tschonoskii maxim alleviates cerebral ischemia-reperfusion injury by inhibiting ferroptosis through Keap-1/Nrf2/HO-1 and Nrf2/SLC7A11/GPX4 pathways
Jian-Hong GAO ; Tian-Ying SONG ; Chao-Xi TIAN ; Fang-Yu ZHAO ; Yi-Duo HE ; Xin LIU ; Xian-Bing CHEN
Chinese Pharmacological Bulletin 2024;40(10):1850-1857
Aim To examine the neuroprotective im-pacts of total saponins from Trillium tschonoskii maxim(TST)on cerebral ischemia-reperfusion injury(CIRI)in rats and delve into the mechanisms of ferroptosis.Methods The CIRI model was prepared by dividing male SD rats into the model group,TST(0.1 g·kg-1)group,Donepezil hydrochloride(0.45 mg·kg-1)group,and sham group.The cognitive functions of rats in each group were assessed through the Morris water maze test,the changes in neurological function were evaluated using the Zea-Longa method,the infarct area was observed via TTC staining,and the pathologi-cal alterations in brain tissue were analysed using HE and Nissl staining.To further investigate the underly-ing mechanism,the mitochondrial structural changes were examined using transmission electron microscopy,and the levels of GSH-PX,MDA,and SOD were ana-lyzed.Additionally,the expressions of GPX4 and Nrf2 proteins were evaluated through immunohistochemistry and immunofluorescence.Furthermore,the protein lev-els of Keap1/Nrf2/HO-1 and Nrf2/SLC7A11/GPX4 pathways in rats were examined using Western blot-ting.Results The rats in the model group displayed diminished learning and memory capabilities in com-parison to those in the sham group,as well as a signifi-cantly increased cerebral infarction area and higher neurological function scores(P<0.01),significantly increased cerebral infarct area,disordered and loosely arranged neurons,and reduced Nissl bodies.Addition-ally,mitochondria showed typical signs of ferroptosis.Changes related to ferroptosis included decreased activ-ities of SOD and GSH-PX(P<0.01)and increased MDA levels(P<0.01).The expression of GPX4 and Nrf2-positive cells was significantly reduced,along with decreased fluorescence intensity of GPX4.Further-more,the protein expression of Keap1,Nrf2,HO-1,GPX4,SLC7A11 in the hippocampus decreased(P<0.05,P<0.01).Following the administration of TST,these effects showed improvement.Conclusions TST has neuroprotective effects,enhancing learning and memory abilities while reducing oxidative stress levels.The mechanism may involve the inhibition of ferroptosis through the Keap-1/Nrf2/HO-1 and Nrf2/SLC7 A11/GPX4 pathways.
2.Effect and complication among different kinds of spinal endoscopic surgery for lumbar disc herniation
Kang CHEN ; Fu-Guo YANG ; Yuan-Chao LUO ; Ren-Jian HE
China Journal of Orthopaedics and Traumatology 2024;37(3):228-234
Objective To compare clinical efficacy and complication rate of percutaneous endoscopic transforaminal discec-tomy(PETD),percutaneous endoscopic interlaminar discectomy(PEID)and unilateral biportal endoscopic(UBE)in treating single-segment lumbar disc herniation(LDH).Methods From October 2019 to August 2021,121 LDH patients with single-segment treated by spinal endoscopy were retrospectively analyzed and divided into three groups.In PETD group,there were 48 patients,including 19 males and 29 females,aged from 18 to 72 years old with an average of(44.0±13.9)years old;3 patients with L3,4 segments,27 patients with L4,5 segments,and 18 patients with L5S1 segments.In PEID group,there were 43 patients,including 23 males and 20 females,aged from 20 to 69 years old with an average of(40.1±12.1)years old;1 patient with L3.4 segments,15 patients with L4.5 segments,and 27 patients with L5S1 segments.In UBE group,there were 30 patients,including 12 males and 18 females,agedfrom29 to 72 years old with an average of(41.2±15.0)years old;1 patient with L3,4 segments,18 patients with L4,5 segments,and 11 patients with L5S1 segments.Operation time,blood loss,fluoroscopy times and complica-tions among three groups were observed and compared.Before opertaion,3 months after operation and at the latest follow-up,visual analogue scale(VAS)was used to evaluate low back pain and lower extremity pain,Oswestry disfunction index(ODI)was used to evaluate lumbar function,and modified MacNab was used to evaluate clinical efficacy at the latest follow-up.Re-sults All patients were performed endoscopic spinal surgery completely and were followed up for at least 12 months.One patient occurred dural sac rupture both in PETD and PEID group,and dural sac rupture was small,and there was no obvious discomfort after operation.Two patients were occurred intraoperative rupture of dural sac in UBE group.One patient was occurred cere-brospinal fluid leakage after operation,and was improved after rest in supine position and fluid rehydration.One patient without no significant postoperative discomfort.(1)There were no significant difference in operating time,blood loss and hospital stay between PETD and PEID group(P>0.05),while UBE group was higher than those of PETD and PEID group(P<0.05).There was no statistical significance in fluoroscopy times between PEID and UBE group(P>0.05),but PETD group was higher than that of PEID and UBE group(P<0.05).(2)VAS of low back pain at 3 months after operation in UBE group was higher than that in PETD and PEID group(P<0.05),but there was no significant difference between PETD and PEID group(P>0.05).At the latest follow-up,there was no significant difference in VAS of low back pain among three groups(P>0.05).(3)Lower ex-tremity pain of VAS and ODI among 3 groups after operation were significantly improved at all time points compared with those before opertaion(P<0.05),and there were no statistical significance between groups(P>0.05),and there were no statistical significance in interaction between different time points and operation groups(P>0.05).(4)At the latest follow-up,according to the modified MacNab standard,the results of PETD group were excellent in 27 patients,good in 16 patients,moderate in 4 patients,poor in 1 patient;in PEID group,27 patients got excellent result,12 good,3 moderate,and 1 poor;in UBE group,16 patients got excellent,10 good,2 moderate,and 2 poor.There was no significant difference among three groups(x2=0.308,P>0.05).Recurrence of lumbar disc herniation occurred in 1 patient among each three groups,symptoms were improved in 2 pa-tients after symptomatic treatment,and 1 patient was treated in other hospitals.Conclusion PETD,PEID and UBE techniques could achieve good early clinical effects in treating lumbar disc herniation with similar complication rates.Both of PETD and PEID are single-channel minimally invasive surgery,with mild intraoperative tissue damage and quick postoperative recovery;while intraoperative fluoroscopy of PETD was relatively more frequent,and PEID was more suitable for L5S1 segment;UBE is a two-channel surgery,in which the intraoperative soft tissue damage is more severe,but exposure is broad,which is more suit-able for complex cases.
3.Two kinds of percutaneous endoscopic lumbar decompression in the treatment of single level lumbar lateral recess stenosis
Kang CHEN ; Yuan-Chao LUO ; Fu-Guo YANG ; Ren-Jian HE
China Journal of Orthopaedics and Traumatology 2024;37(4):338-344
Objective To prospectively compare the clinical efficacy and radiographic outcomes between interlaminar per-cutaneous endoscopic lumbar decompression(IL-PELD)and transforaminar percutaneous endoscopic lumbar decompression(TF-PELD)in the treatment of single-segment lumbar lateral recess stenosis.Methods From April 2018 to July 2021,85 pa-tients with single-segment lumbar lateral recess stenosis underment percutaneous endoscopic lumbar decompression.There were 44 males and 41 females,aged from 49 to 81 years old with an average of(65.5±8.3)years old,duration of lumbar lateral re-cess stenosis ranging from 3 to 83 months with an average of(26.7±16.5)months.They were divided into IL-PELD group and TF-PELD group according to the different operation methods.There were 47 patients in the IL-PELD group,including 28 males and 19 females aged from 50 to 80 yeaes old with an average age was(66.7±9.3)years old.The disease duration ranged from 3 to 65 months with an average of(25.7±15.0)months.There were 38 patients in the TF-PELD group,including 16 males and 22 females,aged from 51 to 78 years old with an average of(64.1±7.6)years old.The disease duration ranged from 4 to 73 months with an average of(27.9±18.3)months The operation time,intraoperative blood loss,intraoperative fluoroscopy,hospi-talization day and complications of the two groups were recorded.Visual analogue scale(VAS)to evaluate low back pain and lower limb pain,Oswestry disability index(ODI)to evaluate lumbar function in preoperative and postoperative period(1 month,6 months and last follow-up)were recorded.the sagittal diameter of the lateral recess of the responsible intervertebral space in preoperative and 1 week after the operation were recorded.Results The operation was successfully completed in both groups without serious complications such as vascular injury,dural sac tear and nerve injury.The operation time in IL-PED group(69.3±19.3)min was significantly longer than that in TF-PELD group(57.5±14.5)min(P<0.05).There was no significant dif-ference in the intraoperative blood loss between the two groups(P>0.05).The number of intraoperative fluoroscopy in TF-PELD group(8.8±2.6)times was significantly higher than that in IL-PED group(4.8±1.2)times(P<0.05).The hospitalization days of the two groups were not statistically significant(P>0.05).VAS for low back and lower extremity pain and ODI were(5.1±2.2),(6.9±1.3)scores and(71.4±12.6)%in IL-PELD group,and(4.7±1.8),(6.9±1.3)scores and(68.4±13.9)%in TF-PELD group.In the IL-PELD group,the VAS of low back pain was(2.4±1.5),(1.6±0.8),(1.4±0.9)scores,and the VAS of lower extremity pain was(3.0±1.2),(1.6±0.7),(1.5±1.0)scores,ODI was(32.6±11.9)%,(17.4±6.5)%,(19.3±9.3)%;In TF-PELD group,the VAS of low back pain was(2.6±1.4),(1.5±0.6),(1.4±1.0)scores,and the VAS of lower extremity pain was(2.8±1.2),(1.6±0.6),(1.5±1.2)scores,The ODI was(32.0±11.2)%,(15.0±6.1)%,and(20.0±11.3)%.The VAS and ODI of the two groups at each time point after operation were significantly improved compared with those before operation(P<0.05),but there was no statistically significant difference between the groups(P>0.05),and there was no statistically sig-nificant difference in the interaction between different time points and groups(P>0.05).At 1 week after operation,the sagittal diameter of lateral recess in both groups was significantly increased compared with that before operation(P<0.05),but there was no significant difference between the two groups at each time point(P>0.05).According to the modified Macnab criteria,IL-PELD group was rated as excellent in 24 cases,good in 19 cases and fair in 4 cases.In TF-PELD group the results were ex-cellent in 19 cases,good in 15 cases,fair in 3 cases and poor in 1 case.There was no significant difference between the two groups(P>0.05).Conclusion IL-PELD and TF-PELD can expand the lateral recess in the treatment of single level lumbar lateral recess stenosis,and have achieved good clinical effects.
4.Percutaneous balloon mitral valvuloplasty guided by intracardiac echocardiography:a report of two cases
De-Jian LI ; Song CHEN ; Chao XU ; Xue JIANG ; Bo WANG ; Jian-Fei FENG ; Dong-Bang SONG ; Guo-Hui ZHANG ; Ming-Quan WANG ; Wei-Min WANG ; Da-Dong ZHANG
Chinese Journal of Interventional Cardiology 2024;32(5):295-297
For the past 30 years,percutaneous balloon mitral valve dilatation has been performed under the guidance of X-rays and bedside ultrasound.However,there are still some cases of mitral valve stenosis in the large atrium where balloon dilation failed.Intraperitoneal ultrasound-guided percutaneous balloon mitral valve plasty is accurate and feasible,which can reduce the occurrence of complications and improve the success rate of such elderly complex cases.Two patients with severe mitral stenosis underwent percutaneous balloon mitral valve plasty guided by intracardiac ultrasound.The operations were successful without any complications,which can provide reference for clinical treatment of mitral stenosis.
5.Protective Effect of Endogenous ω-3 Polyunsaturated Fatty Acid Against Cisplatin-Induced Myelosuppression
Qi-Hua XU ; Zong-Meng ZHANG ; Chao-Feng XING ; Han-Si CHEN ; Ke-Xin ZHENG ; Yun-Ping MU ; Zi-Jian ZHAO ; Fang-Hong LI
Journal of Experimental Hematology 2024;32(5):1601-1607
Objective:To investigate the protective effect of endogenous ω-3 polyunsaturated fatty acid(PUFA)against cisplatin-induced myelosuppression and the mechanism of reducing apoptosis in bone marrow nucleated cells using mfat-1 transgenic mice.Methods:The experimental animals were divided into 4 groups:wild-type mice normal control group,mfat-1 transgenic mice normal control group,wild-type mice model group and mfat-1 transgenic mice model group.The mice in the model group were injected intraperitoneally with 7.5 mg/kg cisplatin on day 0 and day 7 to construct a myelosuppression model,while the mice in the normal control group were injected intraperitoneally with an equal amount of saline,and their status was observed and their body weight was measured daily.Peripheral blood was taken after 14 day for routine blood analysis,and the content and proportion of PUFA in peripheral blood were detected using gas chromatography.Bone marrow nucleated cells in the femur of mice were counted.The histopathological changes in bone marrow were observed by histopathological staining.The apoptosis of nucleated cells and the expression level changes of apoptosis-related genes in the bone marrow of mice were detected by flow cytometry and fluorescence quantitative PCR.Results:Compared with wild-type mice,mfat-1 transgenic mice showed significantly increased levels of ω-3 PUFA in peripheral blood and greater tolerance to cisplatin.Peripheral blood analysis showed that endogenous ω-3 PUFA promoted the recovery of leukocytes,erythrocytes,platelets and haemoglobin in peripheral blood of myelosuppressed mice.The results of HE staining showed that endogenous ω-3 PUFA significantly improved the structural damage of bone marrow tissue induced by cisplatin.Flow cytometry and PCR showed that,compared with wild-type mice model group,the apoptosis rate of bone marrow nucleated cells in mfat-1 transgenic mice was significantly reduced(P<0.001),and the expression of anti-apoptotic genes Bcl-2 mRNA was significantly increased(P<0.01),while the expressions of pro-apoptotic genes Bax and Bak mRNA were significantly reduced(P<0.001,P<0.05).Conclusion:Endogenous ω-3 PUFA can reduce cisplatin-induced apoptosis in bone marrow nucleated cells,increase the number of peripheral blood cells and exert a protective effect against cisplatin-induced myelosuppression by regulating the expression of apoptosis-related genes.
6.A randomized positive parallel-controlled clinical study of Dibai Yijing Formula for the treatment of male infertility with essence deficiency in the kidney and damp-heat type in the essence chamber
Shuang WU ; Chao-Qian CHEN ; Yun-Zhi LI ; Qiu-Jian FENG ; Hui LÜ ; Xiao-Jing AN ; Jun GUO
National Journal of Andrology 2024;30(11):1019-1024
Objective:This study aims to evaluate the clinical efficacy and safety of Dibai Yijing Formula(DYF)in the treat-ment of male infertility with essence deficiency in the kidney and damp-heat in the essence chamber(Abbreviation:kidney deficiency and damp-heat type).Methods:This study employed a randomized,controlled clinical trial design,recruiting 72 male patients with infertility due to kidney deficiency and damp-heat type.Patients were randomly assigned to an treatment group(36 patients)and a control group(36 patients)using a random number table.The control group received oral Clomiphene Citrate Capsules(50 mg,twice daily),while the treatment group received oral DYF(one dose daily,200 ml each time,30 minutes after breakfast and dinner).Both groups underwent a 12-week treatment period.After treatment,sperm concentration(SC),percentage of progressively motile sperms(PR),total sperm motility[PR+percentage of non-progressively motile sperms(NP)],and semen volume(SV)were compared between the two groups before and after treatment.Additionally,the total score of Traditional Chinese Medicine(TCM)syndrome score and sperm DNA fragmentation index(DFI)and pregnancy outcomes of the patients'spouses were compared between the two groups.Results:Three patients dropped out from the treatment group and four from the control group.There were no statistically sig-nificant differences in semen parameters between the two groups(P>0.05).After treatment,the patients in the treatment group showed significant difference in the percentage of SC([19.42±5.30]x 106/ml vs[10.75±2.41]x 106/ml),PR([27.72±6.62]%vs[20.04±4.10]%),PR+NP([49.86±10.68]%vs(33.74±5.58)%],DFI([12.33±3.43]%vs[15.06±3.98]%)and TCM symtom score([7.45±1.82]vs[13.85±1.91]),and the difference was statistically significant(P<0.05).The patients in the control group showed significant difference in the percentage of SC([19.56±5.24]× 106/ml vs[11.31±2.08]× 106/ml)and TCM symptom score([12.81±1.86]vs[14.06±1.64]).But no significant changes were observed in the PR([21.75±5.93]%vs[20.05±4.67]%),PR+NP([34.23±7.15]%vs[32.35±4.09]%),SV([3.19±1.08]ml vs[3.12±1.13]ml),and DFI([15.11±4.76]%vs[15.51±4.35]%)were not statistically significant(P>0.05).Improve-ments in PR,PR+NP,TCM symptom score and DFI in the treatment group were better than those in the control group after treatment(P<0.05);the differences in SC and SV and spousal pregnancy in the treatment group were not statistically significant compared with those in the control group(P>0.05).No serious adverse events occurred in both groups during the treatment period.Conclusion:The treatment of male infertility with DYF is effective and safe.
7.Effects of Moluodan Dami Pills on chronic atrophic gastritis rats
Meng-Lei WANG ; Yi-Feng WU ; Jian-Liang SUI ; Miao-Miao YIN ; Hui-Yun LIU ; Qi-Chao LIU ; Zhuo-Chen WU ; Zhen WANG
Chinese Traditional Patent Medicine 2024;46(5):1476-1482
AIM To investigate the effects of Moluodan Dami Pills on chronic atrophic gastritis(CAG)rats and their mechanism.METHODS The rat models were randomly divided into the model group,the low-dose group and high-dose Moluodan Dami Pills groups(2.43 g/kg and 4.86 g/kg),and vitamin A group(0.32 g/kg),following the 16 weeks successful induction of CAG by five-factor modeling method,in contrast to another 10 normal rats of the control group.After 8 weeks corresponding administration,the rats of each group had their general physiological status and pH value of gastric juice assessed;their pathological changes of gastric mucosa observed by naked eyes combined with HE staining;their changes of gastrin-secreting cells(G cells)and somatostatin-secreting cells(D cells)in gastric mucosa observed by immunohistochemistry;and their serum levels of pepsinogen Ⅰ/pepsinogen Ⅱ(PG Ⅰ/PG Ⅱ)ratio,TNF-α and IL-6 detected by ELISA.RESULTS Compared with the model group,the groups intervened with Moluodan Damei Pills and vitamin A displayed lower pH values of gastric juice(P<0.05),improved pathological changes of gastric mucosa,increased G and D cells counts(P<0.05,P<0.01),increased ratio of serum PGⅠ/PGⅡ,and decreased levels of IL-6 and TNF-α(P<0.05,P<0.01).CONCLUSION Moluodan Dami Pills can effectively improve the symptoms of CAG rats through its influence on the number and secretion abilityof G and D cells,the levels of serum PG Ⅰ/PG Ⅱ ratio and inflammatory factors.
8.Clinical efficacy of AERD in the treatment of residual aortic dissection on Stanford type B aortic dissection patients after endovascular repair
Jinglong ZHANG ; Jia CHEN ; Chao LI ; Zilan SUN ; Zilin LI ; Jian ZUO
Chinese Journal of Thoracic and Cardiovascular Surgery 2024;40(6):331-334
Objective:Evaluate the safety and efficacy of aortic endovascular remodeling device (AERD) in the treatment of residual aortic dissection on Stanford type B aortic dissection patients after endovascular repair.Methods:Adopting a prospective cohort study design, according to the inclusion and exclusion criteria, a total number of 60 patients with Stanford B-type aortic dissection after TEVAR surgery from January 2022 to December 2023 were included. Collecting CTA imaging data of patients before and after endovascular treatment with AERD and using computer 3D reconstruction fluid dynamics to evaluate distal aortic remodeling.Results:The incidence of major adverse events and mortality rate associated with aortic dissection within 30 days were 0 after endovascular treatment with AERD. In the short term, AERD was safe for the treatment of residual aortic dissection on Stanford type B aortic dissection patients after endovascular repair. Endovascular treatment with AERD significantly increased the volume of the true lumen of the aorta in the dissection segment, with an increase of 88.44%. At the same time, endovascular treatment with AERD significantly reduced the volume of the false lumen of the aorta in the dissection segment, with a decrease of 61.04%. Endovascular treatment with AERD has no significant effect on the longest true lumen diameter of distal aorta of stent and upper margin aorta of superior mesenteric artery. Comparing to the longest diameter of the true lumen of the aorta, the change in the shortest diameter of the true lumen of the aorta is more significant, with the shortest diameter of true lumen of distal aortic of the stent increasing by 32.29%, the shortest diameter of true lumen of the narrowest part of the aorta increasing by 204.15%, the shortest diameter of true lumen of the upper margin aorta of superior mesenteric artery increasing by 80.76%, the shortest diameter of true lumen of the lower edge aorta of the renal artery increasing by 115.26%, the shortest diameter of the opening of the left iliac artery increasing by 152.01%, and the shortest diameter of opening of the right iliac artery increasing by144.21%.Conclusion:AERD is safe and effective in the treatment of residual aortic dissection on Stanford type B aortic dissection patients after endovascular repair. Endovascular treatment with AERD promotes true lumen dilation and false lumen reduction in the early postoperative stage and promotes benign remodeling of the distal aorta after TEVAR surgery.
9.Design and experimental study of wearable cardiopulmonary monitoring system
Wan-Jun SHUAI ; Shu-Li ZHAO ; Wen-Zhe LI ; Hua-Yong GAO ; Jian JIANG ; Xi CHEN ; Jin-Hua YANG ; Yong CHAO ; Zheng-Tao CAO
Chinese Medical Equipment Journal 2024;45(4):51-55
Objective To design a wearable cardiopulmonary monitoring system and validate its performance through preliminary human trials.Methods The wearable cardiopulmonary monitoring system was composed of a data collector,a wearing vest and an information management platform.The data collector used an EFM32GG330 SCM as the main microcon-troller unit(MCU),which included a respiratory modulation module,an ECG modulation module,a body position modulation module,a wireless communication module(involving in a Bluetooth module and a Wi-Fi module),a storage module and a power management module.The wearable vest had a cardigan-type structure,and was equipped with ECG sensors and respiratory motion sensors at its inner side.The information management platform was developed with Client/Server(C/S)architecture and Java/JavaScript.The system developed was compared with Mindray's IPM10 Patient Monitor routinely used in hospitals through preliminary human trials to verify its effectiveness in monitoring human heart rate and respiratory rate.Results The system developed could continuously monitor the human heart rate and respiratory rate for a long time,and the monitoring results had high consistency with those of Mindray's IPM10 Patient Monitor.Conclusion The system can be used for medical monitoring of cardiopulmonary indicators during training or exercise,providing accurate physiological information for health management.[Chinese Medical Equipment Journal,2024,45(4):51-55]
10.Expert consensus on the evaluation and rehabilitation management of shoulder syndrome after neek dissection for oral and maxillofacial malignancies
Jiacun LI ; Moyi SUN ; Jiaojie REN ; Wei GUO ; Longjiang LI ; Zhangui TANG ; Guoxin REN ; Zhijun SUN ; Jian MENG ; Wei SHANG ; Shaoyan LIU ; Jie ZHANG ; Jicheng LI ; Yue HE ; Chunjie LI ; Kai YANG ; Zhongcheng GONG ; Qing XI ; Bing HAN ; Huaming MAI ; Yanping CHEN ; Jie ZHANG ; Yadong WU ; Chao LI ; Changming AN ; Chuanzheng SUN ; Hua YUAN ; Fan YANG ; Haiguang YUAN ; Dandong WU ; Shuai FAN ; Fei LI ; Chao XU ; Wei WEI
Journal of Practical Stomatology 2024;40(5):597-607
Neck dissection(ND)is one of the main treatment methods for oral and maxillofacial malignancies.Although ND type is in con-stant improvement,but intraoperative peal-pull-push injury of the accessory nerve,muscle,muscle membrane,fascia and ligament induced shoulder syndrome(SS)is still a common postoperative complication,combined with the influence of radiochemotherapy,not only can cause pain,stiffness,numbness,limited dysfunction of shoulder neck and arm,but also may have serious impact on patient's life quality and phys-ical and mental health.At present,there is still a lack of a systematic evaluation and rehabilitation management program for postoperative SS of oral and maxillofacial malignant tumors.Based on the previous clinical practice and the current available evidence,refer to the relevant lit-erature at home and abroad,the experts in the field of maxillofacial tumor surgery and rehabilitation were invited to discuss,modify and reach a consenusus on the etiology,assessment diagnosis,differential diagnosis,rehabilitation strategy and prevention of SS,in order to provide clinical reference.

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