CN111643510A - Application of anemonin B4 in preparation of medicine for lowering blood pressure - Google Patents
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Abstract
The invention discloses an application of anemonin B4 in preparing a medicine for reducing blood pressure. Pulsatillae saponin B4 has therapeutic effect on hypertension, and has effects of reducing blood pressure of renal hypertension, spontaneous hypertension, protecting hypertension and improving cardiac function.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to application of anemonin B4 in preparation of a medicine for reducing blood pressure.
Background
Abnormal rise of blood pressure is one of the main risk factors for occurrence and development of cardiovascular diseases, and long-term rise of blood pressure or unstable blood pressure causes serious damage to target organs such as heart, brain, kidney, blood vessels and the like, thereby seriously affecting prognosis of diseases. Therefore, the stabilization of blood pressure, the prevention and improvement of the damage of target organs caused by hypertension are the final objective of the treatment of hypertension. Pulsatillae saponin B4 is pentacyclic triterpenoid saponin extracted from Pulsatilla chinensis (Thunb.) nakai of perennial herb of Ranunculaceae, and has antiinflammatory, antibacterial, immunoregulatory, antitumor, antioxidant and antiviral effects, such as: application of Chinese pulsatilla saponin B4 with publication number CN 105213410A as immunomodulator in medicine for treating acute inflammation; the application of the pulsatilla saponin B4 with the publication number of CN 105535004A as an EV71 virus inhibitor in preparing the hand-foot-and-mouth disease resistant medicine is disclosed, but the pulsatilla saponin monomeric compound which has the effect of reducing blood pressure is not reported at present.
Disclosure of Invention
The invention researches the effect of the pulsatilla chinensis saponin B4 in the medicine for treating hypertension from the mechanism of hypertension, and provides scientific basis and experimental basis for preventing and treating hypertension subsequently by pulsatilla chinensis saponin B4.
The invention aims to provide application of anemonin B4 in preparation of a medicine for reducing blood pressure.
Wherein the medicament is a medicament for treating renal hypertension.
Wherein the medicament is a medicament for treating essential hypertension.
The invention at least comprises the following beneficial effects:
pulsatillae saponin B4 has therapeutic effect on hypertension, and has effects of reducing blood pressure of renal hypertension, spontaneous hypertension, protecting hypertension and improving cardiac function.
The pulsatilla saponin B4 is applied to the medicine for treating renal hypertension, can improve the content of Nitric Oxide (NO) in serum, and reduce the content of angiotensin II (Ang II) and Endothelin (ET) in the plasma to improve vasomotor function and reduce blood pressure, and can reduce HW/BW and LVI values of heart, inhibit myocardial hypertrophy, contribute to improving contraction and relaxation functions of heart chambers, and play a role in protecting hypertension and improving heart function.
When the pulsatilla saponin B4 is applied to the medicine for treating the spontaneous hypertension, the Nitric Oxide (NO) content in serum can be increased, the angiotensin II (AngII) content in the plasma can be reduced, the aorta internal diameter can be reduced, and the effects of reducing the blood pressure, protecting the hypertension and improving the cardiac function can be realized.
Experiments in specific embodiments prove the application of the pulsatilla saponin B4 in lowering blood pressure.
Drawings
FIG. 1 is a bar graph of the aortic inner diameters of rats in the placebo, model, B45 mg/kg, B410 mg/kg, B420 mg/kg and closantel potassium groups;
FIG. 2 is a bar graph of the EF values of rats in the placebo, model, B45 mg/kg, B410 mg/kg, B420 mg/kg and closartan potassium tablets groups;
FIG. 3 is a bar graph of the FS values of rats of the blank control group, the model group, the B45 mg/kg group, the B410 mg/kg group, the B420 mg/kg group and the closartan potassium tablet group.
Detailed Description
The present invention is further described in detail below with reference to examples to enable those skilled in the art to practice the invention with reference to the description.
It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.
It is to be noted that the experimental methods described in the following embodiments are all conventional methods unless otherwise specified, and the reagents and materials are commercially available unless otherwise specified.
The data obtained by the experiment are subjected to single-factor variance analysis and t test data by adopting STATA8.0 statistical software, the difference is P <0.05, the difference has statistical significance, and the smaller the P, the better the effect is.
Example 1
Animal experiments of the effect of pulsatilla saponin B4 on renal hypertension were as follows:
1.1 materials
1.1.1 Experimental animals
Healthy male SD rats, 60, SPF grade, body weight (180 ± 20) g, provided by the center of experimental animals at the university of cantonese medical, animal certification number: SCXK Gui 2009-. All experimental animals are raised in a controllable environment at the room temperature of 18-24 ℃ and the humidity of 40-50%, the animals eat and drink water freely during the experiment, and the circadian rhythm is normal.
1.1.2 major drugs and reagents
Pulsatillae saponin B4 (Jiangxi Bencao Tian Gong Tech science and technology, Inc., batch No. 2018042205); captopril (Shandesh, China pharmaceutical Co., Ltd., batch: AK 002); physiological saline for injection (Sichuan Kelun pharmaceutical Co., Ltd., batch No. L219012211); a nitric oxide detection kit (Nanjing institute of bioengineering, lot number: 20190303); angiotensin II ELISA assay kit (Rapiddio, USA, lot: 20190226Q); endothelin ELISA assay kit (Rapiddio, USA, batch: 20190301B).
ALC-NIBP non-invasive blood pressure measurement and analysis system (Australian Coute Biotech, Shanghai); 1/1000 precision balance (Metler-Tollido instruments Shanghai Co., Ltd., model: ME 204E); low temperature high speed centrifuge (Eppendorf Co., model 5425R); 722sp visible spectrophotometer (Shanghai prism technologies, Inc.); an enzyme-linked immunoassay instrument (BioTek company, America, model: SYNERGYH 1).
1.2 Experimental methods
1.2.1 establishment and grouping of animal models
50 healthy male SD rats are fed adaptively for 1 week, the blood pressure of the tail artery of each rat is measured by an ALC-NIBP noninvasive blood pressure determination and analysis system before model building, the measurement is repeated for 3 times, and the average value is taken and recorded. A renal hypertension rat model is prepared by adopting a 'two kidneys and one clamp' method. After 4 weeks of molding, 50 rats meet the systolic blood pressure (SP) which is more than 120mmHg, and the blood pressure after molding is more than 20mmHg higher than that before molding, namely the experimental renal hypertension rats. Taking 50 renal hypertension rats successfully modeled, randomly dividing the renal hypertension rats into 5 groups, namely 10 model control groups (model control groups), 10 positive drug control groups (captopril groups), 10 anemonin B4 high-dose groups, 10 medium-dose groups of anemonin B4 and 10 low-dose groups of anemonin B4; another 10 patients were treated (i.e., free unilateral renal artery, but no constriction, no hypertension formation) (sham control). After 4 weeks after operation, after the SP of the rat is stabilized, the blood pressure of the tail artery of the rat in the waking state is measured by an ALC-NIBP noninvasive blood pressure measurement and analysis system before administration. The pulsatilla chinensis saponin B4 high and medium and low dose groups are 5, 10 and 20mg/kg, the administration is carried out by intraperitoneal injection, the dose of the captopril group is 7.0mg/kg, the administration is carried out by intragastric injection, rats in the model control group and the sham operation control group are injected with physiological saline with the same volume as the rats in the abdominal cavity every day, the administration is carried out 1 time every day from the 5 th week, and the administration is carried out for 4 weeks continuously. Wherein the equivalent volume is the same as the administration volume of the abdominal cavity, namely 1mL/100 g; in the experiment, 7.0mg/kg of captopril is selected as the optimal administration dose, and the captopril is obtained by screening the searched documents and the pre-experimental results.
1.2.2 blood pressure measurement
The tail arterial Systolic Pressure (SP) of the rat in the waking state was measured by ALC-NIBP non-invasive blood pressure measurement and analysis system before, 4 weeks after and 4 weeks after the model was made, and the measurement was repeated 3 times, and the average value was taken, and the obtained data are detailed in Table 1.
Table 1: influence of Pulsatillae saponin B4 on blood pressure of renal hypertensive rat (
mmHg)
Note: the blood pressure of the patients before and after administration is compared,△P<0.05，△△P<0.01; compared with the model control group,#P<0.05，##P<0.01。
the data in table 1 represent the effect of pasqueflower saponin B4 on the blood pressure in renal hypertensive rats. The method specifically comprises the following steps: the mean blood pressure of the rats before molding was 106.34mmHg, the mean blood pressure of the rats after 4 weeks of molding was 151.15mmHg, and the blood pressure change was significantly different (P <0.01) compared with the sham control group (107.72 mmHg). After 4 weeks of administration, the pulsatilla saponin B4 high-medium low-dose group and the captopril group both generate obvious blood pressure lowering effects, and have significant difference (P <0.01) compared with a model control group, and compared with the self blood pressure before and after administration, the pulsatilla saponin B4 high-medium low-dose group (5mg/kg, 10mg/kg and 20mg/kg) and the captopril group both have significantly lower blood pressure than the blood pressure before administration, which indicates that the pulsatilla saponin B4 and the captopril have significant blood pressure lowering effects. The data in table 1 clearly show that the pulsatilla saponin B4 has a better blood pressure lowering effect when applied to the medicine for treating renal hypertension.
1.2.3 detection of hematological indicators
Continuously administering for 4 weeks, performing 10% chloral hydrate intraperitoneal injection anesthesia (3.0mL/kg) 12h after the last administration, and collecting blood via carotid artery intubation, EDTA-K2The anticoagulation tube is used for blood collection for separating plasma, and the vacuum blood collection tube (without anticoagulation) is used for blood collection for separating serum. EDTA-K2Centrifuging the anticoagulation tube blood sample by a low-temperature centrifuge at 4000r/min for 10min to separate plasma, storing the blood sample in a refrigerator at minus 80 ℃ for later use, placing the vacuum blood collection tube blood sample in a constant-temperature water tank at 37 ℃, standing for 30min to 50min, centrifuging the blood sample by the low-temperature centrifuge at 4 ℃ at 4000r/min for 10min after the serum is separated out, separating the serum, and storing the blood sample in the refrigerator at minus 80 ℃ for later use. (1) determining the content of serum NO by a colorimetric method, and operating the experimental steps according to the instruction of the NO detection kit. (2) The content of Ang II in blood plasma is determined by an enzyme-linked immunoassay method, and the experimental steps are operated according to the instruction of an Ang II ELISA determination kit. (3) The ET content of the plasma is determined by an enzyme-linked immunosorbent assay, and the experimental steps are operated according to the ET ELISA kit instruction. The data obtained by the detection are detailed in table 2.
Table 2: influence of Pulsatillae saponin B4 on serum NO and plasma Ang II and ET of renal hypertension rat
Note: compared with the model control group,#P<0.05，##P<0.01。
the data in Table 2 show the effect of Pulsatillae saponin B4 on the serum NO and plasma Ang II and ET content of renal hypertensive rats. The method specifically comprises the following steps: compared with a model control group, the serum NO content of the pulsatilla saponin B420 mg/kg group and the captopril group is obviously increased (P <0.01 or P <0.05), the plasma AngII content of the pulsatilla saponin B410 mg/kg group and the captopril group is obviously reduced (P <0.01), and the plasma ET content of the pulsatilla saponin B45 mg/kg group and the captopril group is obviously reduced (P < 0.01). The data in Table 2 show that the pulsatilla saponin B4 and the captopril can improve the NO content in serum and reduce the Ang II and ET content in the plasma, and are beneficial to improving the vasomotor function, protecting hypertension and reducing the blood pressure.
1.2.4 cardiac index determination
The rats were sacrificed and the hearts were weighed for total Heart Weight (HW) using a precision balance, the atria and right ventricles were trimmed off, the left ventricles (including the ventricular septum) were retained, weighed to obtain the Left Ventricular Weight (LVW), and the heart index (HW/BW) was calculated as total heart weight/body weight and the left ventricular weight index (LVI) as left ventricular weight/body weight. The data obtained by the measurement are detailed in table 3.
Table 3: influence of Pulsatillae saponin B4 on HW/BW and LVI values of renal hypertensive rat
Note: compared with the model control group,#P<0.05，##P<0.01; compared with the group of the pseudo-operation,△P<0.05，△△P<0.01。
the data in table 3 represent the effect of pasqueflower saponin B4 on the cardiac index of renal hypertensive rats. The method specifically comprises the following steps: after four weeks of administration, when the rats were sacrificed and the hearts were removed, the volume of the hearts of the rats in the model control group was increased and the ventricular wall muscle layer was thickened compared with that of the sham operation control group, and the ventricular wall muscle layer of each of the administered groups of pulsatilla saponin B4 was thinner than that of the model control group. Compared with the model control group, the HW/BW (heart weight/body weight) and LVI of each dose group and captopril group of the pulsatilla saponin B4 are reduced compared with the model control group. The data in Table 3 show that the pulsatilla saponin B4 can reduce HW/BW and LVI values of the heart, inhibit myocardial hypertrophy, improve the contraction and relaxation functions of ventricles and protect hypertension.
1.3 conclusion
The experimental result of the embodiment 1 shows that the pulsatilla saponin B4 has the effects of reducing the blood pressure of the renal hypertension rats, increasing the content of Nitric Oxide (NO) in serum, reducing the content of angiotensin II (Ang II) and Endothelin (ET) in the plasma to improve the vasomotor function and reduce the blood pressure, reducing the HW/BW and LVI values of the heart, inhibiting myocardial hypertrophy, and contributing to improving the contraction and relaxation functions of the ventricle and protecting the hypertension.
Example 2
Animal experiments of the effect of pulsatilla saponin B4 on spontaneous hypertension were as follows:
2.1 materials
2.1.1 Experimental animals
Healthy male 9-week-old Spontaneously Hypertensive Rats (SHR)50, 9-week-old male wistar kyoto rats (WKY)10, SPF grade, body weight (180 ± 20) g, provided by the experimental animals center of the university of medical science in guangxi, animal certification number: SCXK Gui 2009-. All experimental animals are raised in a controllable environment at the room temperature of 18-24 ℃ and the humidity of 40-50%, the animals eat and drink water freely during the experiment, and the circadian rhythm is normal.
2.1.2 major drugs and reagents
Pulsatillae saponin B4 (Jiangxi Bencao Tian Gong Tech science and technology, Inc., batch No. 2018042205); losartan potassium tablet (Hangzhou Moshadong pharmaceutical Co., Ltd., batch No. L016172); physiological saline for injection (Sichuan Kelun pharmaceutical Co., Ltd., lot number: L219012211); a nitric oxide detection kit (Nanjing institute of bioengineering, lot number: 20190303); angiotensin II ELISA assay kit (Rapiddio, USA, lot: 20190226Q).
ALC-NIBP non-invasive blood pressure measurement and analysis system (Australian Coute Biotech, Shanghai); 1/1000 precision balance (Metler-Tollido instruments Shanghai Co., Ltd., model: ME 204E); low temperature high speed centrifuge (Eppendorf Co., model 5425R); 722sp visible spectrophotometer (Shanghai prism technologies, Inc.); enzyme-linked immunoassay instrument (BioTek company, USA, model: SYNERGYH 1); small animal ultrasonic detectors (Visualsonics, Vev 770).
2.2 Experimental methods
2.2.1 animal grouping and administration
50 healthy male SHR rats 9 weeks old are fed adaptively for 1 week and are randomly divided into 5 groups, namely 10 model control groups, 10 positive drug control groups (losartan potassium tablets), 10 pulsatilla saponin B4 high-dose groups, 10 pulsatilla saponin B4 medium-dose groups and 10 pulsatilla saponin B4 low-dose groups; the blank group contained 10 WKY rats as a control group. The pulsatilla chinensis saponin B4 high, medium and low dose groups are administrated by intraperitoneal injection once a day at 5, 10 and 20mg/kg, the closantine potassium tablet group is ground and then administrated by intragastric administration once at 31.5mg/kg a day, and rats in the model group and the blank control group are administrated by intraperitoneal injection of physiological saline with the same volume for 1 time a day and are continuously administrated for 10 weeks. Wherein the equivalent volume is the same as the administration volume of the abdominal cavity, namely 1mL/100 g; in the experiment, 31.5mg/kg of losartan potassium tablets is selected as the optimal administration dose, and the losartan potassium tablets are obtained by searching documents and screening the results of preliminary experiments.
2.2.2 blood pressure measurement
The tail artery Systolic Pressure (SP) of the rat in the awake state was measured 3 times a week using an ALC-NIBP non-invasive blood pressure measurement analysis system before and 10 weeks after the administration, and the data obtained by averaging the values are described in Table 4.
Table 4: influence of Pulsatillae saponin B4 on blood pressure of spontaneously hypertensive rat (
mmHg)
Note: compared with the blank control group, the composition of the composition,**P<0.01，***P<0.001; in comparison with the set of models,△P<0.05，△△P<0.01，△△△P<0.001; compared with the losartan potassium tablet group,#P<0.05，##P<0.01，###P<0.001。
the data in table 4 represent the effect of pasqueflower saponin B4 on blood pressure in spontaneously hypertensive rats. The method specifically comprises the following steps: after 1 week of administration, the mean blood pressure of the model group is obviously increased compared with that of the blank control group (P < 0.001); the mean blood pressure of the losartan potassium group is obviously reduced compared with that of the model group (P < 0.001); compared with the model group, the pulsatilla saponin B4 administration group has no obvious antihypertensive effect; as the administration time was prolonged, the losartan potassium and anemonin high dose groups exhibited stable blood pressure lowering effects compared to the model group until after the fifth week. The data in table 4 show that pasqueflower saponin B4 also has a blood pressure lowering effect on spontaneous hypertension.
2.2.3 detection of hematological indicators
After the administration is finished, 10% chloral hydrate is anesthetized by intraperitoneal injection (3.0mL/kg) 12h after the last administration, and blood is taken through a carotid artery cannula, and EDTA-K2The anticoagulation tube is used for blood collection for separating plasma, and the vacuum blood collection tube (without anticoagulation) is used for blood collection for separating serum. EDTA-K2Centrifuging the anticoagulation tube blood sample by a low-temperature centrifuge at 4000r/min for 10min to separate plasma, storing the plasma in a refrigerator at minus 80 ℃ for later use, placing the vacuum blood collection tube blood sample in a constant-temperature water tank at 37 ℃, and standing for 30min to 50minAnd min, after the serum is separated out, centrifuging for 10min at 4 ℃ and 4000r/min by using a low-temperature centrifuge, separating the serum, and storing in a refrigerator at minus 80 ℃ for later use. (1) determining the content of serum NO by a colorimetric method, and operating the experimental steps according to the instruction of the NO detection kit. (2) The content of Ang II in blood plasma is determined by an enzyme-linked immunoassay method, and the experimental steps are operated according to the instruction of an Ang II ELISA determination kit. The data detected are shown in Table 5.
Table 5: influence of Pulsatillae saponin B4 on serum NO and plasma Ang II of spontaneous hypertensive rat
Note: compared with the model control group,#P<0.05，##P<0.01。
table 5 shows the effect of Pulsatillae saponin B4 on serum NO and plasma Ang II content of spontaneously hypertensive rats. The method specifically comprises the following steps: compared with the model group, the serum NO content of the pulsatilla saponin B45 mg/kg group and the losartan potassium group is remarkably increased (P <0.01 or P <0.05), and the plasma Ang II content of the pulsatilla saponin B45 mg/kg group and the losartan potassium group is remarkably reduced (P <0.01 or P < 0.05). The data in Table 5 show that the anemonin B4 can improve the NO content in serum and reduce the Ang II content in plasma, and is helpful for reducing blood pressure, protecting hypertension and improving cardiac function.
2.2.4 echocardiography testing
After weighing the rats, anesthetizing the rats by using 1% sodium pentobarbital according to the ratio of 8mL/kg, then slightly scraping the precordial hairs by using a blade, coating the rats with a coupling agent, and then detecting the rats by using a small animal ultrasonic detector. Placing the probe in front of the left chest and pointing to the upper right of the left chest to obtain a long-axis section of the left ventricle; the diameter of the root of the left ventricular outflow tract aorta in diastole is measured by a ruler, and the aorta inner diameter, EF and FS values are respectively measured by taking a short-axis section beside the mitral valve position, and the detailed values are shown in a bar chart of the aorta inner diameter of each group of rats in figure 1, a bar chart of the EF value of each group of rats in figure 2 and a bar chart of the FS value of each group of rats in figure 3, wherein P is less than 0.05, P is less than 0.01, P is less than 0.001, and the asterisks on the bar chart indicate that the statistical significance is achieved.
Referring to the bar graphs of the aortic inner diameters of rats of the blank control group, the model group, the group B45 mg/kg, the group B410 mg/kg, the group B420 mg/kg and the losartan potassium group in FIG. 1, it is shown that the aortic structures and functions are affected by long-term hypertension, and the aortic inner diameters of spontaneous hypertension rats of the model group in the experiment are obviously widened (P <0.001) compared with the blank control group; compared with the model group, the aortic inner diameter values of the pulsatilla saponin B45 mg/kg group and the above dose group and losartan potassium group are reduced (P is less than 0.001). The data in fig. 1 illustrate that pulsatillae radix saponin B4 can lower the inner diameter of aorta, contributing to the protection of hypertension.
EF. FS is an important index for evaluating left ventricular function, see the bar graph of the EF values of rats in the blank control group, model group, B45 mg/kg group, B410 mg/kg group, B420 mg/kg group and losartan potassium group of fig. 2, and the bar graph of the FS values of rats in the blank control group, model group, B45 mg/kg group, B410 mg/kg group, B420 mg/kg group and losartan potassium group of fig. 3 show that the EF and FS values of the model group are significantly reduced (P <0.001) compared with the blank control group; compared with the model group, the EF and FS values of the pulsatilla saponin B45 mg/kg group and the dosage groups above and the losartan potassium tablet group are obviously improved. The data in fig. 2 and 3 demonstrate that pulsatillae radix saponin B4 can improve the heart function of spontaneously hypertensive rats.
2.3 conclusion
The experimental result of the embodiment 2 shows that the pulsatilla saponin B4 has the effects of reducing the blood pressure of spontaneous hypertensive rats, increasing the content of Nitric Oxide (NO) in serum, reducing the content of angiotensin ii (Ang ii) in plasma, reducing the inner diameter of aorta, and increasing the EF and FS values, thereby achieving the effects of reducing the blood pressure and improving the cardiac function.
The experimental results of the embodiment 1 and the embodiment 2 show that the pulsatilla saponin B4 has a therapeutic effect on hypertension, reduces the blood pressure of renal hypertension rats and spontaneous hypertension rats, increases the NO content in the serum of the renal hypertension rats, reduces the content of Ang II and ET in the plasma, reduces the HW/BW and LVI values of the heart, inhibits myocardial hypertrophy, contributes to improving the contraction and relaxation functions of ventricles, and plays a role in protecting the hypertension; can also improve the NO content in the blood serum of a spontaneous hypertensive rat, reduce the Ang II content in the high blood plasma, reduce the inner diameter of the aorta, improve the heart function of the spontaneous hypertension, increase the EF and FS values and improve the heart function of the spontaneous hypertensive rat. In conclusion, the pulsatilla chinensis saponin B4 has obvious effect in the medicine for treating hypertension through the action mechanism, can reduce the blood pressure of renal hypertension and spontaneous hypertension, and has the effects of protecting hypertension and improving cardiac function.
The number of apparatuses and the scale of the process described herein are intended to simplify the description of the present invention.
While embodiments of the invention have been disclosed above, it is not limited to the applications set forth in the description and the embodiments, which are fully applicable in a variety of fields of endeavor to which the invention pertains, and further modifications may readily be made by those skilled in the art, it being understood that the invention is not limited to the details shown and described herein without departing from the general concept defined by the appended claims and their equivalents.
Claims (3)
1. The anemonin B4 can be used for preparing medicine for lowering blood pressure.
2. The use of anemonin B4 in the preparation of a medicament for lowering blood pressure according to claim 1, wherein the medicament is a medicament for treating renal hypertension.
3. The use of anemonin B4 in the preparation of a medicament for lowering blood pressure according to claim 1, wherein the medicament is a medicament for treating essential hypertension.
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| CN201911054468.8A CN110623971A (en) | 2019-10-31 | 2019-10-31 | Application of anemonin B4 in preparation of medicine for lowering blood pressure |
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| CN111643510A true CN111643510A (en) | 2020-09-11 |
| CN111643510B CN111643510B (en) | 2023-03-14 |
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| CN105213410A (en) * | 2015-10-20 | 2016-01-06 | 刘琦 | Anemoside B4 is as the application of immunomodulator in treatment acute inflammation medicine |
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- 2019-10-31 CN CN201911054468.8A patent/CN110623971A/en not_active Withdrawn
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- 2020-03-31 CN CN202010243790.1A patent/CN111643510B/en active Active
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