Magnetic resonance imaging combined with serum endolipin and galactagoglobin-3 to diagnose cerebral infarction in the elderly with diabetes mellitus

BACKGROUND Magnetic resonance imaging(MRI)combined with serum endothelin and galactagoglobin-3(Gal-3)can improve the clinical diagnosis of diabetes mellitus complicated with cerebral infarction.AIM To analyze the clinical value of MRI combined with serum endolipin and Gal-3 for the diagnosis of cerebral infarction in the elderly with diabetes mellitus.METHODS One hundred and fifty patients with acute cerebral infarction hospitalized between January 2021 and December 2023 were divided into two groups according to comorbid diabetes mellitus,including 62 and 88 cases in the diabetic and nondiabetic cerebral infarction groups.Serum samples were collected to detect the expression of serum endolipoxins,and Gal-3,and cranial MRI was performed at admission.Differences between the two groups were compared to analyze the diagnostic value of these parameters.RESULTS Serum endolipin and Gal-3 expression were higher in the diabetic cerebral infarction group(P<0.05).The arterial wall area,vessel area,normalized wall index,and lumen stenosis rate were higher in the diabetic cerebral infarction group,while the rate of arterial lumen moderate and severe stenosis was 48.39% higher(36.36%,P<0.05).The percentage of large(29.03%)and multiple infarcts(33.87%)in the diabetic cerebral infarction group was higher(13.64% and 20.45%),and the incidence rate of lacunar infarcts was lower(37.10%vs 65.91%)(P<0.05).The total incidence of arterial plaque in patients in the diabetic cerebral infarction group was 96.77% higher(69.32%),while the incidence of necrotic lipid core plaque was 58.06%higher(26.14%)(P<0.05).Receiver operating characteristic curve analysis was performed to assess the diagnosis utility of these techniques.MRI in combination with serum endoglin and Gal-3 had the highest area under the curve,the Yoden index,sensitivity and specificity(P<0.05).CONCLUSION The expression of serum endolipin and Gal-3 in elderly patients with diabetes mellitus with cerebral infarction showed an elevated trend,and the degree of luminal stenosis was severe.MRI predominantly revealed large and multiple infarct foci.This combined index examination can improve the clinical diagnosis of diabetes mellitus combined with cerebral infarction.

Magnetic resonance diffusion tensor imaging following major ozonated autohemotherapy for treatment of acute cerebral infarction

Major ozonated autohemotherapy has been shown to promote recovery of upper limb motor function in patients with acute cerebral infarction, but whether naajor ozonated autohelnotherapy affects remote in）ury remains poorly understood. Here, we assumed that major ozonated autohemotherapy contributes to recovery of clinical function, possibly by reducing remote injury after acute cerebral infarction. Sixty acute cerebral infarction patients aged 30-80 years were equally and randomly allocated to ozone treatment and control groups. Patients in the ozone treatment group received medical treatment and major ozonated autohemotherapy （47 mg/L, 100 mL ozone） for 10 ± 2 days. Patients in the control group received medical treatment only. National Institutes of Health Stroke Scale score, modified Rankin scale score, and reduced degree of fractional anisotropy values of brain magnetic resonance diffusion tensor imaging were remarkably decreased, brain function improved, clinical efficiency significantly increased, and no obvious adverse reactions detected in the ozone treatment group compared with the control group. These findings suggest that major ozonated autohemotherapy promotes recovery of neurological function in acute cerebral infarction patients by reducing re,note injury, and additionally, exhibits high safety.

Remodeling of motor cortex function in acute cerebral infarction patients following human urinary kallidinogenase A functional magnetic resonance imaging evaluation after 6 months

A total of 29 patients were treated within 48 hours after acute subcortical cerebral infarction with Xuesaitong or Xuesaitong plus human urinary kallidinogenase for 14 days. Neurological deficits, activity of daily living, and evaluations of distal upper limb motor functions at the 6-month follow-up showed that patients treated with Xuesaitong plus human urinary kallidinogenase recovered better than with Xuesaitong alone. In addition, functional MRI revealed that activation sites were primarily at the ipsilesional side of injury in all patients. Human urinary kallidinogenase induced hyperactivation of the ipsilesional primary sensorimotor cortex, premotor cortex, supplementary motor area, and contralesional posterior parietal cortex. Results showed that human urinary kallidinogenase improved symptoms of neurological deficiency by enhancing remodeling of long-term cortical motor function in patients with acute cerebral infarction.

Metabolic Changes in Rats with Photochemically Induced Cerebral Infarction and the Effects of Batroxobin:A Study by Magnetic Resonance Imaging,~1H-and ^(31)p-Magnetic Resonance Spectroscopy

Metabolic changes in rats with photochemically induced cerebral infarction and the effects of batroxobin were investigated 1, 3, 5 and 7 days after infarction by means of magnetic resonance imaging (MRI), 1H- and 31P-magnetic resonance spectroscopy (MRS). A region of T2 hyperintensity was observed in left temporal neocortex in infarction group and batroxobin group 1, 3, 5 and 7 days after infarction. The volume of the region gradually decreased from 1 day to 7 days after infarction. The ratio of NAA/Cho + Cr in the region of T2 hyperintensity in the infarction group was significantly lower than that in the corresponding region in the sham-operated group 3, 5 and 7 days after infarction respectively (P

Magnetic resonance perfusion imaging evaluation in perfusion abnormalities of the cerebellum after supratentorial unilateral hyperacute cerebral infarction

Magnetic resonance imaging （MRI） data of 10 patients with hyperacute cerebral infarction （≤6 hours） were retrospectively analyzed.Six patients exhibited perfusion defects on negative enhancement integral maps,four patients exhibited perfusion differences in pseudo-color on mean time to enhance maps,and three patients exhibited perfusion differences in pseudo-color on time to minimum maps.Dynamic susceptibility contrast-enhanced perfusion weighted imaging revealed a significant increase in region negative enhancement integral in the affected hemisphere of patients with cerebral infarction.The results suggest that dynamic susceptibility contrast-enhanced perfusion weighted imaging can clearly detect perfusion abnormalities in the cerebellum after unilateral hyperacute cerebral infarction.

Consistency between magnetic resonance diffusion-weighted images and pathological findings in a hyperacute cerebral infarction rabbit model

BACKGROUND： Because magnetic resonance diffusion-weighted imaging is sensitive to water molecule movement, it has particular advantages for early diagnosis of cerebral infarction. However, the relationship between apparent diffusion coefficient changes with ischemia time, particularly relative apparent diffusion coefficient and tissue pathological changes remains controversial. OBJECTIVE： To explore the correlation between apparent diffusion coefficient changes and pathologic changes in hyperacute cerebral infarction. DESIGN, TIME AND SETTING： A randomized, controlled, animal experiment of neuroimaging. The study was performed at the Laboratory of Radiology Department, Longgang Central Hospital of Shenzhen from October 2007 to October 2008. MATERIALS： Magnetic resonance scanner was purchased from Philips Medical Systems, Best, the Netherlands. METHODS： A total of 42 healthy, adult, New Zealand rabbits were randomly assigned into sham-operation, ischemia 0.5-, 1-, 2-, 3-, 4-, and 6-hour groups, with six animals in each group. Local cerebral ischemia model was established by right middle cerebral artery occlusion, and cranial MRI scanning and pathologic observation were performed, respectively, at 0.5, 1,2, 3, 4, and 6 hours following ischemia. The middle cerebral artery of sham-operation group was only exposed, but not occluded. Images at the above-mentioned time points were also collected. MAIN OUTCOME MEASURES： Apparent diffusion coefficient and relative apparent diffusion coefficient values of abnormal signal on diffusion-weighted imaging were calculated and compared with pathological changes in the ischemic region. RESULTS： No abnormal diffusion-weighted imaging signals or pathological changes were observed in the sham-operation group. Abnormal signal intensity on diffusion-weighted imaging was first observed in the 0.5-hour group. Apparent diffusion coefficient and relative apparent diffusion coefficient values decreased in all middle cerebral artery occlusion rabbits and reached lowest levels at 3 hours, followed by a gradual increase. The right ischemic basal ganglia region with high signal intensity on diffusion-weighted imaging extended with increasing time of occlusion, and the pathologic outcome corresponded with MRI changes. CONCLUSION： Relative apparent diffusion coefficient values changed regularly with ischemia time and displayed good correspondence to pathological manifestations.

Neuroimaging diagnosis for cerebral infarction An 8-year bibliometric analysis

OBJECTIVE： To identify global research trends in neuroimaging diagnosis for cerebral infarction using a bibliometric analysis of the Web of Science. DATA RETRIEVAL： We performed a bibliometric analysis of data retrieval for neuroimaging diagnosis for cerebral infarction containing the key words ＂CT, magnetic resonance imaging, MRI, transcranial Doppler, transvaginal color Doppler, digital subtraction angiography, and cerebral infarction＂ using the Web of Science. SELECTION CRITERIA： Inclusion criteria were： （a） peer-reviewed articles on neuroimaging diagnosis for cerebral infarction which were published and indexed in the Web of Science; （b） original research articles and reviews; and （c） publication between 2004-2011. Exclusion criteria were： （a） articles that required manual searching or telephone access; and （b） corrected papers or book chapters. MAIN OUTCOME MEASURES： （1）Annual publication output; （2） distribution according to country; （3） distribution according to institution; （4） top cited publications; （5） distribution according to journals; and （6） comparison of study results on neuroimaging diagnosis for cerebral infarction. RESULTS： Imaging has become the predominant method used in diagnosing cerebral infarction. The most frequently used clinical imaging methods were digital subtraction angiography, CT, MRI, and transcranial color Doppler examination. Digital subtraction angiography is used as the gold standard. However, it is a costly and time-consuming invasive diagnosis that requires some radiation exposure, and is poorly accepted by patients. As such, it is mostly adopted in interventional therapy in the clinic. CT is now accepted as a rapid, simple, and reliable non-invasive method for use in diagnosis of cerebrovascular disease and preoperative appraisal. Ultrasonic Doppler can be used to reflect the hardness of the vascular wall and the nature of the plaque more clearly than CT and MRI. CONCLUSION： At present, there is no unified standard of classification of cerebral infarction imaging. Detection of clinical super-acute cerebral infarction remains controversial due to its changes on imaging, lack of specificity, and its similarity to a space-occupying lesion. Neuroimaging diagnosis for cerebral infarction remains a highly active area of research and development.

Three-dimensional arterial spin labeling and diffusion kurtosis imaging in evaluating perfusion and infarct area size in acute cerebral ischemia

BACKGROUND Early thrombolytic therapy is crucial to treat acute cerebral infarction,especially since the onset of thrombolytic therapy takes 1-6 h.Therefore,early diagnosis and evaluation of cerebral infarction is important.AIM To investigate the diagnostic value of magnetic resonance multi-delay threedimensional arterial spin labeling(3DASL)and diffusion kurtosis imaging(DKI)in evaluating the perfusion and infarct area size in patients with acute cerebral ischemia.METHODS Eighty-four patients who experienced acute cerebral ischemia from March 2019 to February 2021 were included.All patients in the acute stage underwent magnetic resonance-based examination,and the data were processed by the system’s own software.The apparent diffusion coefficient(ADC),average diffusion coefficient(MD),axial diffusion(AD),radial diffusion(RD),average kurtosis(MK),radial kurtosis(fairly RK),axial kurtosis(AK),and perfusion parameters post-labeling delays(PLD)in the focal area and its corresponding area were compared.The correlation between the lesion area of cerebral infarction under MK and MD and T2-weighted imaging(T2WI)was analyzed.RESULTS The DKI parameters of focal and control areas in the study subjects were compared.The ADC,MD,AD,and RD values in the lesion area were significantly lower than those in the control area.The MK,RK,and AK values in the lesion area were significantly higher than those in the control area.The MK/MD value in the infarct lesions was used to determine the matching situation.MK/MD<5 mm was considered matching and MK/MD≥5 mm was considered mismatching.PLD1.5s and PLD2.5s perfusion parameters in the central,peripheral,and control areas of the infarct lesions in MK/MD-matched and-unmatched patients were not significantly different.PLD1.5s and PLD2.5s perfusion parameter values in the central area of the infarct lesions in MK/MD-matched and-unmatched patients were significantly lower than those in peripheral and control areas.The MK and MD maps showed a lesion area of 20.08±5.74 cm^(2) and 22.09±5.58 cm^(2),respectively.T2WI showed a lesion area of 19.76±5.02 cm^(2).There were no significant differences in the cerebral infarction lesion areas measured using the three methods.MK,MD,and T2WI showed a good correlation.CONCLUSION DKI parameters showed significant difference between the focal and control areas in patients with acute ischemic cerebral infarction.3DASL can effectively determine the changes in perfusion levels in the lesion area.There was a high correlation between the area of the infarct lesions diagnosed by DKI and T2WI.

Multimodal magnetic resonance imaging for assessing lacunar infarction after proximal middle cerebral artery occlusion in a canine model

Background A new lacunar infarction model was recently established in beagle dogs through proximal middle cerebral artery （MCA） occlusion by thrombus. This study aimed to characterize the model by multimodal magnetic resonance imaging （MRI） and to investigate its potential role for the future stroke research. Methods The left proximal MCA was embolized with an autologous thrombus in six beagles. Diffusion-weighted imaging （DWI） and T2-weighted imaging （T2Wl） were performed every half hour during the first six hours after occlusion, followed by three time points at 12 hours, 24 hours, and one week. Perfusion-weighted imaging （PWI） and magnetic resonance angiography （MRA） were carried out at six hours, 24 hours and one week. The PWI-DWI mismatch ratio was defined as （PWI-DWl）/DWl ischemic volume. Results Lacunar infarcts induced by MCA occlusion were located in the left caudate nucleus and internal capsule. All the lesions could be detected within two hours by DWI. Lesion volume on DWl increased in a time dependent manner, from （87.19±67.16） mm3 at one hour up to （368.98±217.05） mm3 at 24 hours （P=0.009）, while that on PWl gradually decreased from （7315.00±2054.38） mm3at six hours to （4900.33±1319.71） mm3 at 24 hours and （3334.33±1195.11） mm3 at one week （P=0.002）. The mismatch ratio was 41.93±22.75 at six hours after ischemia, showing ＂extensive mismatch＂, and decreased to 18.10±13.74 at 24 hours （P=0.002）. No MCA recanalization was observed within 24 hours after MCA occlusion. Conclusions Lacunar infarction induced by proximal MCA occlusion could be detected early by DWl and was characterized by extensive PWI-DWl mismatch. Multimodal MRI is useful to demonstrate the natural evolution of PWI-DWl mismatch. This ischemic model could be further used for investigating early thrombolysis in lacunar stroke showing extensive mismatch.

脑桥梗死锥体束继发Wallerian变性的磁共振扩散张量成像研究

目的应用磁共振扩散张量成像(DTI)研究脑桥梗死后锥体束继发Wallerian变性的演变过程与神经功能恢复的关系。方法对16例脑桥梗死患者(病例组)于发病<7 d、15 d、30 d、90 d和180 d进行DTI检查。另选16例正常志愿者作为对照组。测量梗死区下方的延髓、上方的大脑脚、内囊后肢及中央前后回皮质的各向异性分数(FA)值,比较病例组各时间点梗死侧与对侧以及对照组各部位FA值的差异,计算FA比值(rFA),并进行临床神经功能评分分析。结果与对照组比较,病例组脑桥梗死灶同侧的延髓、大脑脚、内囊后肢以及中央前后回皮质FA值均有不同程度降低,这种降低的趋势在发病15 d时最为显著,180 d基本恢复正常。患者发病15d时的rFA与90d、180d时的Fugl-Meyer运动功能评分均呈正相关。结论 DTI能够早期检测脑桥梗死后锥体束纤维继发的Wallerian变性。发病早期病灶同侧锥体束FA值下降的程度可预示患者远期运动功能恢复的程度。

脑梗死后锥体束Wallerian变性的DTI研究进展

运动通路缺血性脑卒中可导致病灶远端的锥体束发生Wallerian变性,扩散张量成像技术显示Wallerian变性不仅具有较高的敏感性和特异性,而且能在活体定量监测其演变过程,这对判断脑卒中患者的预后具有重要意义。本文主要针对脑卒中后锥体束Wallerian变性的扩散张量研究进展进行综述。

磁共振弥散张量成像对Wallerian变性诊断价值的评价

目的:探讨磁共振弥散张量成像技术(diffusion tensor imaging,DTI)在Wallerian变性中的诊断价值。方法:对40例正常志愿者、10例正常变异者和40例病变者分别进行DTI和常规MRI扫描。结果:正常志愿者大脑脚、脑桥基底部皮质脊髓束走行区各向异性分数(fractional anisotropy,FA)均值分别为0.73和0.63,其FA比值(数值小的一侧比大的一侧)分别为97.6%和96.8%;正常变异者大脑脚、脑桥基底部皮质脊髓束走行区FA均值分别为0.72和0.64,其FA值比值分别为96%和95%,与正常志愿者相比,差异无统计学意义(P>0.05);病变者患侧大脑脚、脑桥基底部FA均值分别为0.64及0.52。其FA比值(患侧FA值比健侧FA值)分别为76.0%和77.0%,患侧大脑脚、脑桥基底部皮质脊髓束的FA值及其比值较正常者明显减小,差异有统计学意义(P<0.05)。结论:DTI技术通过测量FA值及其比值及神经纤维束的三维重建,较常规MRI更为准确客观地诊断Wallerian变性。

脑梗塞后锥体束Wallerian变性的MRI表现

目的分析脑梗塞后锥体束Wallerian变性的MRI表现及其临床意义。方法回顾性分析15例经常规MRI扫描发现有Wallerian变性的脑梗塞病人的临床及MRI资料。结果常规MRI在脑梗塞后1-2周也能发现典型的Wallerian变性,Wallerian变性的出现与梗塞的大小有关,Wallerian变性越重,出现的越早,病人的预后越差。结论Wallerian变性的观察有助于临床医师对病人肢体功能恢复的评估。

脑损伤后锥体束Wallerian变性的氢质子磁共振波谱成像

目的:探讨氢质子磁共振波谱(1HMRS)成像技术检测和定量分析脑损伤后锥体束Wallerian变性的可行性。方法:对15例单侧脑损伤且出现同侧锥体束Wallerian变性患者和15例年龄匹配的健康志愿者(对照组)行常规MRI检查,并同时行1HMRS检查,测量双侧大脑脚锥体束走行区的NAA/Cr、Cho/Cr值,比较该值在脑损伤组与对照组间的差异。结果:脑损伤后同侧大脑脚谱线图显示NAA峰减低,Cho峰升高,NAA/Cr值较对侧明显减低,Cho/Cr值明显升高,与对侧及对照组比较,差异均存在统计学意义(P<0.001),而病变组对侧大脑脚及对照组双侧大脑脚的NAA/Cr、Cho/Cr值差异无统计学意义。结论:1HMRS技术可以检测并定量分析脑损伤后锥体束Wallerian变性。

Wallerian变性的MRI诊断

目的 :研究Wallerian变性的MRI表现 ,提高诊断正确率。方法 :结合原发性脑损伤的发生情况 ,对 2 7例Wallerian变性的MRI资料进行分析。结果 :2 7例中 7例原发性脑损伤后 15～ 2 4周患者 ,Walleri an变性分别表现为同侧放射冠、内囊后肢、大脑脚和脑桥基底部的T2 WI高信号区 ;2 0例原发性脑损伤后 1.5～ 7年患者 ,Wallerian变性表现为同侧脑干萎缩。结论

脑梗死后皮质脊髓束Wallerian变性的DTI纵向研究

目的探讨应用扩散张量成像（DTI）观察脑梗死后从早期、亚急性至慢性期同侧皮质脊髓束（CST）的DTI参数随时间变化的规律，由此检测皮质脊髓束发生Wallerian变性的时间演变过程。方法选取2008年11月至2010年1月间在复旦大学附属中山医院住院治疗的皮层下运动通路脑梗死患者34例，发病时间从5h至2年。在入院时行MRI＋DTI检查。34例中选取5例患者，在发病1W内、第2周、1个月、3个月、6个月和1年分别行MRI＋DTI检查。采用工作站之Functool软件，重建出平均弥散系数（MD）、各向异性分数（FA）的参数图。通过放置兴趣区（ROI）的方式计算出FA值、MD值、rFA、rMD。ROI放置在双侧大脑脚。结果脑梗死后病灶同侧大脑脚区rFA在1月内下降最明显，1个月至3个月下降变慢，3个月后rFA变化幅度很小基本保持平稳。而rMD在脑梗死2W内无明显变化，2W至3个月上升，3个月后基本保持稳定。5例患者不同时间点的DTI研究能够显示这种变化过程。结论脑梗死后从早期、亚急性至慢性期同侧大脑脚处皮质脊髓束的FA值进行性减少。DTI有利于检测脑梗死后皮质脊髓束发生Wallerian变性的程度及动态演变过程。

常规MR诊断Wallerian变性的临床价值

目的 探讨常规MRI在诊断脑损害后锥体束Wallerian变性中的价值。方法 对69例脑损害后锥体束Wallerian变性患者行MRI检查。常规行快自旋回波序列（FSE常规横断位、矢状位及冠状位扫描）：1.5T：T1WI FSE（TR550ms TE 15ms）T2WI FSE（TR 2 800ms TE 131ms）0.5T：T1WI SE（TR 500ms,TE 15ms）;T2WI FSE（TR 3 000ms,TE130ms）,层厚5mm,层间距1.0mm。结果 69例Wallerian患者中,Wallerian患者脑损害发生时间与检查时间为5周-3年,其中4-5周2例,5周-12月60例,大于12月7例,异常信号经放射冠、穿过内囊膝部及（或）后肢的前半部、大脑脚外侧及脑桥基底部达延髓锥体,且连续多于3个层面（3个层面以上）可观察到;常规MRI条状异常信号为稍长T1信号,长T2信号2例,T2Flair序列呈斑片状稍高信号;67例条状异常信号为长T1长T2信号,有12例观察到病侧脑干萎缩,以中脑患侧大脑脚萎缩较显著。结论 MRI可明确显示脑损害后锥体束Wallerian变性,有助于临床医师对患者预后正确评估及制定运动功能康复的治疗计划。

FA值在脑梗死及其继发Wallerian变性中的临床意义

目的 探讨FA值对脑梗死及其继发Wallerian变性的临床诊断价值.方法 收集脑梗死患者32例,所有患者均分别在发病后7天内、3～4周进行DTI检查,测量梗死区、健侧相应部位脑组织、双侧大脑脚的FA值,对比检查结果.结果 ①脑梗死灶、患侧大脑脚较对侧相应部位FA值明显下降,差异有统计学意义（P＜0.05）；②脑梗死灶、患侧大脑脚FA值均呈逐渐下降趋势,前后两次FA测量值差异均有统计学意义（P＜0.05）；③健侧对照区脑组织、健侧大脑脚前后两次FA值变化均无统计学意义（P＞0.05）.结论 FA值在脑梗死及其继发Wallerian变性的病理进程中逐渐降低,在Wallerian变性的早期诊断及患者运动功能预后判断中有重要临床意义.

中枢神经系统Wallerian变性MR影像特征分析

目的分析多种原因引起的锥体束wallerian变性的MRI影像学特征。方法分析34例我院MRI发现的wallerian变性患者的影像学特点,总结统计学结果。结果在原发病变基础上,可见继发病变均沿锥体束方向分布;MRI表现为等或长T_1、等或稍高T_2信号,FLAIR呈稍高信号,早期DWI呈高信号,中晚期DWI呈等信号;变性晚期均显示伴有患侧大脑脚及桥脑萎缩。结论中枢神经系统多种原发病可累及锥体束导致锥体束继发性Wallerian变性,影像科医师结合wallerian变性的病变特点及MRI影像特点,可以提高对该病的诊断。

Offspring of rats with cerebral hypoxia-ischemia manifest cognitive dysfunction in learning and memory abilities

Neonatal hypoxic-ischemic encephalopathy is a serious neurological disease,often resulting in long-term neurodevelopmental disorders among surviving children.However,whether these neurodevelopmental issues can be passed to offspring remains unclear.The right common carotid artery of 7-day-old parental-generation rats was subjected to permanent ligation using a vessel electrocoagulator.Neonatal hypoxic-ischemic rat models were established by subjecting the rats to 8%O2–92%N2 for 2 hours.The results showed that 24 hours after hypoxia and ischemia,pathological damage,cerebral atrophy,liquefaction,and impairment were found,and Zea-Longa scores were significantly increased.The parental-generation rats were propagated at 3 months old,and offspring were obtained.No changes in the overall brain structures of these offspring rats were identified by magnetic resonance imaging.However,the escape latency was longer and the number of platform crossings was reduced among these offspring compared with normal rats.These results indicated that the offspring of hypoxic-ischemic encephalopathy model rats displayed cognitive impairments in learning and memory.This study was approved by the Animal Care&Welfare Committee of Kunming Medical University,China in 2018(approval No.kmmu2019072).