The present application claims the priority of the prior applications in China with the Application Nos. CN202211569444.8; CN202211569445.2; CN202211569450.3; and CN202211581572.4 filed on Dec. 8, 2022. The description, claims, and drawings of the description incorporated by reference herein are incorporated in their entirety.
The present disclosure belongs to the technical field of biomaterials and particularly relates to a composite decellularized matrix membrane for preventing calculi and use thereof.
An incidence of bladder cancer is the first named among urological malignancies and increases year by year. Researchers perform a sampling survey of patients who died due to malignant tumors. The result shows that the number of the dead due to bladder cancer is first ten. Bladder cancer poses a serious threat to life quality and health of patients and can be classified into muscle-invasive bladder cancer and non-muscle-invasive bladder cancer depending on whether the tumor invades a muscle layer, wherein the muscle-invasive bladder cancer is a fatal disease. Radical total cystectomy, lymph node dissection, and permanent urinary diversion are a main surgery mode for treating recurrent and multiple invasive bladder cancer at present, are also a first choice method for treating muscle-invasive bladder cancer, and can effectively reduce recurrent metastasis and improve a survival rate. However, the surgery is difficult and time-consuming, and has many complications after the surgery. The method destroys a normal structure and function of a digestive tract of a patient, also has many complications such as intestinal obstruction, metabolism disorder or nutrition disorder, mucus secretion in an artificial bladder, inflammation, calculus formation, etc. after the surgery, and seriously affects life quality of the patient after the surgery.
A non-crosslinked extracellular matrix-based biomaterial uses an animal small intestine submucosa (SIS) tissue as a raw material, such that risks such as immunogenicity and the like are removed, and a structure and active ingredients of a natural extracellular matrix are retained. After the material is implanted into a body, the material endogenously induces regeneration and repair, realizes tissue function regeneration, can be completely degraded in a body, has advantages of inflammation tolerance, adhesion prevention and the like, and is almost suitable for the regeneration and repair of all soft tissues of human body.
In order to solve the above problems of a bladder surgery, it is urgent to find a composite decellularized matrix membrane for preventing calculi.
The present disclosure provides a composite decellularized matrix membrane for preventing calculi and use thereof. The composite decellularized matrix membrane prepared by the present disclosure is prepared by wrapping one layer of a polymer membrane with two layers of decellularized matrix membranes. The composite decellularized matrix membrane has effects on preventing water infiltration and calculi, makes a patient suffer less, recover faster, live better, and cost less, while medical resources are saved, and hope for treatment is also brought for some patients with a contraindication to an in-situ ileal neobladder surgery.
To solve the above problems, the present disclosure uses the following technical solutions.
A composite decellularized matrix membrane for preventing calculi comprises a decellularized matrix membrane and a polymer membrane. The composite decellularized matrix membrane is formed by wrapping one layer of a polymer membrane with two layers of decellularized matrix membranes.
The decellularized matrix membrane is prepared from at least a biomaterial of a porcine small intestine, a porcine bladder, and porcine skin. The polymer membrane is prepared from at least a polymer material of polycaprolactone, polylactic acid, and polyurethane.
Preferably, the decellularized matrix membrane is prepared from a porcine small intestine. Further, the polymer membrane is prepared from polycaprolactone. In some embodiments, the polymer membrane is prepared from polylactic acid. In some embodiments, the polymer membrane is prepared from polyurethane. Use of the composite decellularized matrix membrane in the preparation of an artificial bladder for preventing water infiltration. The composite decellularized matrix membrane comprises the composite decellularized matrix membrane in the above embodiment. Use of the composite decellularized matrix membrane in the preparation of an absorbably degradable artificial bladder for preventing water infiltration. The composite decellularized matrix membrane comprises the composite decellularized matrix membrane in the above embodiment. Specifically, when the decellularized matrix membrane is prepared from a porcine small intestine and the polymer membrane is polycaprolactone, polylactic acid or polyurethane, the composite decellularized matrix membrane is usable for preparing a water infiltration-preventing and/or absorbably degradable artificial bladder.
In some embodiments, the decellularized matrix membrane is prepared from at least a biomaterial of a porcine small intestine, a porcine bladder, and porcine skin. The polymer membrane is prepared from at least a polymer material of polycaprolactone, polydimethylsiloxane, polyurethane, polylactic acid-glycolic acid, polyvinyl alcohol, and polyhydroxy fatty acid ester.
Preferably, the decellularized matrix membrane is prepared from a porcine small intestine and the polymer membrane is prepared from polycaprolactone. In some embodiments, the polymer membrane is prepared from polyurethane. In some embodiments, the polymer membrane is prepared from polyvinyl alcohol. In some embodiments, the polymer membrane is prepared from polyhydroxy fatty acid ester. Use of the composite decellularized matrix membrane in the preparation of an artificial bladder for avoiding formation of calculi. The composite decellularized matrix membrane comprises the composite decellularized matrix membrane in the above embodiment. Specifically, when the polymer membrane is polycaprolactone, polydimethylsiloxane, polyurethane, polylactic acid-glycolic acid, polyvinyl alcohol, and polyhydroxy fatty acid ester, the composite decellularized matrix membrane is usable for preparing an artificial bladder for avoiding formation of calculi.
Preferably, when the decellularized matrix membrane is prepared from a porcine small intestine and the polymer membrane is polycaprolactone or polyurethane, the composite decellularized matrix membrane is usable for preparing an absorbably degradable artificial bladder for preventing calculi and water infiltration. The “preventing calculi” mentioned above refers to not producing calculi in a bladder patch, repair or bladder replacement surgery.
In some embodiments, the decellularized matrix membrane is prepared by the following steps:
A method for preparing the composite decellularized matrix membrane comprises the following steps:
The mold in step (2) is a balloon or a contractive spherical support.
A mass-volume ratio of the polymer material to the solvent in step (1) is (1-10):100. Preferably, the polymer material is selected from polycaprolactone, further, the solvent is selected from chloroform, and a preferable mass-volume ratio is 1:100.
In one aspect, the present disclosure provides a composite decellularized matrix membrane with a good toughness, wherein the composite decellularized matrix membrane comprises a decellularized matrix membrane and a polymer membrane, and the composite decellularized matrix membrane is formed by wrapping one layer of the polymer membrane with two layers of the decellularized matrix membranes.
In some embodiments, the decellularized matrix membrane is prepared from at least a biomaterial of a porcine small intestine, a porcine bladder, and porcine skin.
In some embodiments, the polymer membrane is prepared from at least a polymer material of polycaprolactone, polydimethylsiloxane, polyurethane, polylactic acid-glycolic acid, polyether-ether-ketone, and polystyrene.
In some embodiments, the decellularized matrix membrane is prepared from a porcine small intestine.
In some embodiments, the polymer membrane is prepared from polycaprolactone.
In some embodiments, the polymer membrane is prepared from polyurethane.
In some embodiments, the polymer membrane is prepared from polyether-ether-ketone.
In some embodiments, the polymer membrane is prepared from polystyrene.
In another aspect, the present disclosure provides a composite decellularized matrix membrane for preventing inflammation, wherein the composite decellularized matrix membrane comprises a decellularized matrix membrane and a polymer membrane, and the composite decellularized matrix membrane is formed by wrapping one layer of the polymer membrane with two layers of the decellularized matrix membranes.
In some embodiments, the decellularized matrix membrane is prepared from at least a biomaterial of a porcine small intestine, a porcine bladder, and porcine skin.
In some embodiments, the polymer membrane is prepared from at least a polymer material of polycaprolactone, polydimethylsiloxane, polyurethane, polylactic acid-glycolic acid, polyvinyl alcohol, and polyhydroxy fatty acid ester.
In some embodiments, the decellularized matrix membrane is prepared from a porcine small intestine.
In some embodiments, the polymer membrane is prepared from polycaprolactone.
In some embodiments, the polymer membrane is prepared from polydimethylsiloxane.
In some embodiments, the polymer membrane is prepared from polyvinyl alcohol.
In some embodiments, the polymer membrane is prepared from polyhydroxy fatty acid ester.
In some embodiments, the composite decellularized matrix membrane comprises the composite decellularized matrix membrane according to claims 1-8.
In another aspect, the present disclosure provides a composite decellularized matrix membrane for preventing deformation, wherein the composite decellularized matrix membrane comprises a decellularized matrix membrane and a polymer membrane, and the composite decellularized matrix membrane is formed by wrapping one layer of the polymer membrane with two layers of the decellularized matrix membranes, wherein the decellularized matrix membrane is prepared from at least a biomaterial of a porcine small intestine, a porcine bladder, and porcine skin.
In some embodiments, the polymer membrane is prepared from at least a polymer material of polycaprolactone, polyalkylcyanoacrylate, polylactic acid-glycolic acid, and L-polylactic acid.
In some embodiments, the decellularized matrix membrane is prepared from a porcine small intestine.
In some embodiments, the polymer membrane is prepared from polytetrafluoroethylene.
In some embodiments, the polymer membrane is prepared from polypropylene.
In some embodiments, the polymer membrane is prepared from poly(N-vinylpyrrolidone).
In some embodiments, the polymer membrane is prepared from polyethylene or polyvinylpyrrolidone.
In some embodiments, the present disclosure provides use of the composite decellularized matrix membrane in the preparation of a ureter for preventing inflammation. The composite decellularized matrix membrane comprises the aforementioned composite decellularized matrix membrane. The polymer membrane is prepared from at least a polymer material of polycaprolactone, polyalkylcyanoacrylate, polylactic acid-glycolic acid, and L-polylactic acid.
The present disclosure has the following beneficial effects:
In order to describe the present disclosure more specifically, the technical solutions of the present disclosure will be described in detail below in combination with the drawings and specific embodiments. These descriptions only show how the present disclosure is realized and do not limit the specific scope of the present disclosure. The scope of the present disclosure is defined by the claims.
The decellularized matrix membrane provided by the present example comprised the following preparation steps:
The decellularized matrix membrane provided by the present example comprised the following preparation steps:
The decellularized matrix membrane provided by the present example comprised the following preparation steps:
The decellularized matrix membrane provided by the present example comprised the following preparation steps:
The decellularized matrix membrane provided by the present example comprised the following preparation steps:
The decellularized matrix membrane provided by the present example comprised the following preparation steps:
The decellularized matrix membrane provided by the present example comprised the following preparation steps:
The decellularized matrix membrane provided by the present example comprised the following preparation steps:
The decellularized matrix membrane provided by the present example comprised the following preparation steps:
The decellularized matrix membrane provided by the present example comprised the following preparation steps:
The composite decellularized matrix membrane provided by the present example comprised the following preparation steps as shown in
The composite decellularized matrix membrane provided by the present example comprised the following preparation steps:
The composite decellularized matrix membrane provided by the present example comprised the following preparation steps:
The composite decellularized matrix membrane provided by the present example comprised the following preparation steps:
The composite waterproof balloon provided by the present example comprised the following preparation steps:
The composite waterproof balloon provided by the present example comprised the following preparation steps:
The composite waterproof balloon provided by the present example comprised the following preparation steps:
The preparation steps of the present comparative example were the same as those of comparative example 1. A difference was that the polymer material is L-polylactic acid.
Test samples: spherical composite decellularized matrix membranes prepared in examples 6 and 7 and comparative examples 1 and 2;
Test method: a non-testing end was blocked, artificial urine at a temperature of 37+/−2° C. was filled into the spherical composite decellularized matrix membrane, the spherical composite decellularized matrix membrane was placed at a room temperature for 4 h, whether liquid seeps or not on a surface of the spherical composite decellularized matrix membrane was observed by naked eyes, and whether liquid seeps or not on an outer surface of the spherical composite decellularized matrix membrane was wiped by hands shown in
meanwhile, another composite decellularized matrix membrane was taken, a non-testing end was blocked, artificial urine at a temperature of 37+/−2° C. was filled in the spherical composite decellularized matrix membrane, a pressure value was applied to a testing end and at least kept for 12 h, whether the sample leaks or not was visually inspected, and test results were shown in the following table.
Testing results: A water infiltration-preventing performance of the membrane in examples 6 and 7 was better than that of the membrane in comparative examples 1 and 2. Besides, the membrane in examples 6 and 7 can bear a positive pressure pressurization of 8 KPa at most, while a critical value capable of being borne by a human bladder was 40 cm of a water column, namely about 4 KPa. The water infiltration-preventing performance of the membrane in examples 6 and 7 can reach a standard of the human bladder.
Test samples: composite decellularized matrix membrane prepared in example 5.1.
Rabbit bladder surgery: (1) anesthesia: an experimental rabbit was anesthetized by an intraperitoneal injection with sodium pentobarbital (40 mg/kg) at a concentration of 1%.
(2) Skin preparation: hairs on an abdomen of the experimental rabbit were removed in a sterile environment and the skin was cleaned and disinfected with 2% iodophor.
(3) A wound with a length of 7-8 cm was cut on the abdominal skin of the experimental rabbit, a bladder was found out, a wound with a length of 1 cm was cut on the bladder, and a sample was cut to a proper size and attached to the wound to be sutured.
(4) Activities and feeding of the experimental rabbit were observed periodically and the experimental rabbit was dissected 1 month after the surgery for a degradation study.
Porcine bladder surgery: (1) 30 min before a surgery, a pig was intramuscularly injected with 0.05 mg/kg of atropine, 0.1 mg/kg of midazolam and 5 mg of a morphine hydrochloride injection, the pig was in a supine position with four limbs fixed, an ear vein was punctured, and 5% of a glucose injection was dripped into an abdomen at a lower abdominal center incision.
(2) An incision was 15-20 cm long. Skin at two sides of the incision was fixed by using a thumb and a forefinger of a surgeon, a tip of a scalpel was vertically punctured into the skin, then the scalpel was rotated to a 45° oblique angle of a skin surface, the skin and a tissue 5 cm below the skin were uniformly incised by using the scalpel, and the scalpel was rotated to be 90° to a vertical direction to a skin surface and taken out; if a cut length of a subcutaneous tissue was shorter than that of the skin, the subcutaneous tissue can be cut by scissors; and a cutting force should be suitable to cut out the skin at one time, such that an incision edge was neat and linear, and uneven edges caused by multiple times of incision to influence healing were avoided.
(3) Subcutaneous fat, muscle, and fascia were isolated. The sample was clamped by two hemostatic forceps and lifted upwards by a surgeon and cut between the two forceps. A proper force was exerted. A tip of the scalpel was always upward to avoid injuring deep organs and tissues. Abdominal wall muscles at 0.5 cm away from left and right sides of a center line of an abdominal wall were respectively clamped by using toothed forceps respectively, the scalpel was used vertically to cut a small opening, and the abdominal wall muscles were cut along the center line.
(4) Sterile gauze was soaked by normal saline and then spread near the incision, such that a subsequent operation was convenient, a bladder was reached, and a wound of 2-3 cm of the bladder was longitudinally cut.
(5) Prepared bladder support materials were respectively sewn on the experimental animal, whether the materials were firm or not was checked, a 0-4 micro suture was used, continuous suture and discontinuous overlock were performed, retention catheterization was performed after a repair, and a water injection test was performed.
(6) Continuous suture was performed layer by layer with 2-0 silk thread and abdominal wall muscles and skin were closed. Finally, skin incision was adjusted by using toothed forceps for folding and the skin was sterilized by iodophor for 2 times.
(7) An inhalation of an anesthetic was stopped after the surgery was finished, the experimental animal was given a full oxygen breathing, and after the experimental animal was awake, a tracheal catheter was removed after the experimental animal recovered a spontaneous breathing.
(8) Activities and feeding of the experimental pig were observed periodically and the experimental pig was dissected 2 month after the surgery for a degradation study.
Testing results: a condition of the experimental rabbit 1 month after the surgery was shown in
Pathological figures of the bladder of the experimental rabbit were shown in
A suture condition of the experimental pig was shown in
Pathological sections of specimens taken 2 months after the surgery were shown in
In conclusion, the rabbit bladder surgery and the porcine bladder surgery both showed that the decellularized matrix membrane prepared by the present disclosure can prevent water infiltration and was absorbably degradable when used for a bladder repair and a total bladder repair. Predictably, in a clinical practice, when the composite decellularized matrix membrane provided by the present disclosure can be degraded, absorbed, and free of residue, a healed bladder was mainly an autologous tissue.
A preparation method of the composite decellularized matrix membrane provided by the present example comprised the following steps:
The preparation steps and the amount of the polymer of the example were basically the same as those in example 8 except that polylactic acid-glycolic acid was selected as a polymer material and chloroform was selected as a solvent.
The preparation steps and the amount of the polymer of the example were basically the same as those in example 8 except that polydimethylsiloxane was selected as a polymer material and chloroform was selected as a solvent.
The preparation steps and the amount of the polymer of the example were basically the same as those in example 8 except that polyvinyl chloride was selected as a polymer material and cyclohexanone was selected as a solvent.
The preparation steps and the amount of the polymer of the example were basically the same as those in example 8 except that polyvinyl alcohol was selected as a polymer material and chloroform was selected as a solvent.
The preparation steps and the amount of the polymer of the example were basically the same as those in example 8 except that polyhydroxy fatty acid ester was selected as a polymer material and chloroform was selected as a solvent.
The preparation steps and the amount of the polymer of the example were basically the same as those in example 8 except that polyether-ether-ketone was selected as a polymer material and chloroform was selected as a solvent.
The preparation steps and the amount of the polymer of the example were basically the same as those in example 8 except that polyacrylonitrile was selected as a polymer material.
The preparation steps and the amount of the polymer of the example were basically the same as those in example 8 except that polystyrene was selected as a polymer material and dimethyl sulfoxide was selected as a solvent.
Test samples: the composite decellularized matrix membranes prepared in examples 5.1 and 8-16.
Test method: the test method was the same as the porcine bladder surgery.
Test results: postoperative conditions of the composite decellularized matrix membranes prepared from SIS with PU, PCL, PLGA, PDMS, PVA, and PHA polymer materials were shown in
Postoperative conditions of the composite decellularized matrix membranes prepared from SIS with PVC, PEEK, PAN, and PS polymer materials were shown in
Test samples: the composite decellularized matrix membrane prepared in example 5.1.
Test method: normal urinary tract epithelial cells SVHUC-1 transfected with a GFP fluorescent protein were inoculated on a bladder patch repaired by using the composite decellularized matrix membranes prepared in example 5.1, after the cells were adhered to a wall for 8 h, the cells were fully rinsed for 3 times, then adherent cells were desorbed, and a fluorescence intensity was measured.
Testing results: as shown in
Test samples: the composite decellularized matrix membrane prepared in example 5.1.
Test method: a medium cultured with a bladder patch repaired using the composite decellularized matrix membrane prepared in example 5.1 for a certain period of time (day 1, day 3, day 6, and day 9) was cultured together with normal urinary tract epithelial cells SVHUC-1 transfected with a GFP fluorescent protein for 24 h. A cell activity was examined.
Test results: the cell activity was shown in
Test samples: the composite decellularized matrix membranes prepared in examples 5.1 and 8-16.
Test method: the test method was the same as the porcine bladder surgery.
Test results: calculus formation of the composite decellularized matrix membranes prepared from SIS with PVC, PEEK, PAN, and PS polymer materials was shown in
Test samples: the composite decellularized matrix membranes prepared in examples 5.1 and 8-16.
Test method: the composite decellularized matrix membrane was folded at 180° or folded again.
Test results: a qualified toughness standard was that an artificial bladder was folded at 180° in any direction and then folded at 180° again in the same direction, an artificial bladder should be recovered, and defects such as folding or rupture and the like should not occur in an artificial bladder body by visual inspection.
The test results of the composite decellularized matrix membranes prepared from SIS with PU, PCL, PLGA, PDMS, PEEK, and PS polymer materials were shown in
The test results of the composite decellularized matrix membranes prepared from SIS with PVC and PHA polymer materials were shown in
The test results of the composite decellularized matrix membranes prepared from SIS with PVA and PAN polymer materials were shown in
The above experimental results were specifically shown in the following table:
Result analysis: when the composite decellularized matrix membranes prepared from the decellularized matrix membrane and PCL, PU, PLGA, and PDMS used for a bladder patch, inflammation and calculus formation did not occur. Meanwhile, after folded at 180° in any direction and then folded at 180° again in the same direction, the artificial bladder can restore to an original shape and it was determined that defects such as folding or rupture and the like did not occur according to a folded appearance. An inflammation phenomenon and calculus formation did not occur, indicating that the artificial bladder prepared from the composite decellularized matrix membrane provided by the present disclosure in clinical practice did not enable a patient to be like a patient subjected to a sigmoid colorectal cystectomy who easily suffered from serious complications such as urinary tract inflammation, water electrolyte acid-base equilibrium disorder, even intestinal tumors and the like. The artificial bladder had few complications, did not damage an intestinal tract, and greatly reduced surgical risks and patient injury. A qualified toughness indicated that after the composite decellularized matrix membrane provided by the present disclosure was used for the bladder surgery, the membrane had a lower distortion resistance and good deformation performance in a human body, no folding and rupture fracture occurred due to deformation of the composite membrane, urine leakage was prevented, patch suturing was not affected during bladder contracture, foreign body sensation did not easily occur after the surgery, and the membrane had good stability.
A preparation method of the tubular composite decellularized matrix membrane provided by the present example comprised the following steps:
The preparation steps of the example were basically the same as those in example 17 except that polyalkylcyanoacrylate (PACA) was selected as a polymer material.
The preparation steps of the example were basically the same as those in example 17 except that polylactic acid-glycolic acid (PLGA) was selected as a polymer material.
The preparation steps of the example were basically the same as those in example 17 except that L-polylactic acid (PLLA) was selected as a polymer material.
The preparation steps of the example were basically the same as those in example 17 except that polyethylene was selected as a polymer material.
The preparation steps of the example were basically the same as those in example 17 except that polytetrafluoroethylene (PTFE) was selected as a polymer material.
The preparation steps of the example were basically the same as those in example 17 except that polypropylene (PP) was selected as a polymer material.
The preparation steps of the example were basically the same as those in example 17 except that polycarbonate (PC) was selected as a polymer material.
The preparation steps of the example were basically the same as those in example 17 except that polyvinylpyrrolidone (PVP) was selected as a polymer material.
The preparation steps of the example were basically the same as those in example 17 except that poly(N-vinylpyrrolidone) (PNVP) was selected as a polymer material.
Test samples: composite decellularized matrix membranes prepared in examples 17-26.
Test method: the test method was the same as the porcine bladder surgery. A test part was replaced into a ureter.
Test results: the test results of the composite decellularized matrix membranes prepared from SIS with PACA, PCL, PLGA, PLLA, and PTEF polymer materials were shown in
A postoperative condition of the composite decellularized matrix membranes prepared from SIS with PE, PP, PC, PVP, and PNVP polymer materials were shown in
Results of a postoperative stenosis test of the composite decellularized matrix membranes prepared from SIS with PACA, PCL, PLGA, PLLA, PTEF, PP, and PNVP polymer materials were shown in
Test samples: composite decellularized matrix membranes prepared in examples 17-26.
Test method: a test device shown in
Test results: acceptance criteria were that when a force of 1 N was applied on a product, a deflection should be less than or equal to 25 mm and a deformation lasting for 1 min should be less than or equal to 2.5 mm. Experimental results of a ureter were shown in the following table:
Result analysis: when the composite decellularized matrix membranes prepared from SIS with PCL, PACA, PLGA, and PLLA polymer materials used in a clinical experiment of the ureter, inflammation and stenosis did not occur, rejection reaction brought by an allogeneic tissue and ureteral hydrops and hydronephrosis caused by postoperative stenosis were avoided, and a problem of histocompatibility easily caused by an artificial material replacement was solved. Occurrences of complications of a patient were avoided, and surgical risks and pains of the patient were greatly relieved. Meanwhile, rigidity and strength of the membrane enable the membrane to have a certain supporting force in a body of the patient. In a normal floating range of the ureter, no contracture occurred and perforation or displacement deformation was avoided.
All the patents and publications mentioned in the description of the present disclosure indicate that these are public technologies in the art and can be used by the present disclosure. All the patents and publications cited herein are listed in the references, just as each publication is specifically referenced separately. The present disclosure described herein can be realized in the absence of any one element or multiple elements, one restriction or multiple restrictions, where the limitation is not specifically described here. For example, in each example, the terms “comprise”, “substantially composed of” and “composed of” can be replaced by the remaining two terms of either. The so-called “a” here only means “a kind”, not excluding only one, but also can indicate two or more. The terms and expressions used herein are descriptive, without limitation. Besides, there is no intention to indicate that these terms and interpretations described in the description exclude any equivalent features. However, it can be known that any appropriate changes or modifications can be made within the scope of the present disclosure and claims. It can be understood that the examples described in the present disclosure are some preferred examples and features. A person skilled in the art can make some modifications and changes according to the essence of the description of the present disclosure. These modifications and changes are also considered to fall within the scope of the present disclosure and the scope limited by independent claims and dependent claims.
Number | Date | Country | Kind |
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2022115694448 | Dec 2022 | CN | national |
2022115694452 | Dec 2022 | CN | national |
2022115694503 | Dec 2022 | CN | national |
2022115815724 | Dec 2022 | CN | national |