CN2661199Y - Microcarrier stirred type biological artificial liver support system - Google Patents
Microcarrier stirred type biological artificial liver support system Download PDFInfo
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- CN2661199Y CN2661199Y CN 200320109050 CN200320109050U CN2661199Y CN 2661199 Y CN2661199 Y CN 2661199Y CN 200320109050 CN200320109050 CN 200320109050 CN 200320109050 U CN200320109050 U CN 200320109050U CN 2661199 Y CN2661199 Y CN 2661199Y
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- blood plasma
- bioreactor
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Abstract
The utility model relates to a medical instrument, in particular to a bioartificial liver support system that temporarily substitutes the human liver. The utility model comprises a biological reactor with a substance exchanging function, an incubator with the function of a stirring incubation bottle, a plasma/ incubation liquid constant flow device, a slow magnetic stirrer, a constant temperature device (an air supplying and control device of CO2 and O2), a membrane oxygenator and a plasma pool. The utility model is the bioartificial liver support system, which is combined with the mechanical artificial liver, and can substitute the liver function of hepatic failure patient and support liver transplantation for the severe hepatic failure patient or help the hepatic failure patient gradually recover hepatic tissue and regeneration and compensation functions, thereby enhancing the survival rate and prolonging the survival time of the hepatic failure patient.
Description
Technical field:
This utility model relates to a kind of armarium, particularly a kind of temporary replacement human liver's biological artificial liver support system.Be specifically related to that a kind of and mechanical artificial liver is used in combination, can substitute liver failure patient liver function in a long time, support the severe liver patient that declines to wait for liver transplantation, perhaps make the liver failure patient recover self hepatic tissue and its regeneration and compensation gradually, patient's survival rate and the biological artificial liver support system of prolongation time-to-live thereby the raising liver declines.
Background technology:
Liver failure is common critical illness.China is " hepatitis big country ", and the average year sickness rate is about 1,00/,100,000, and promptly national annual acute viral hepatitis is about 1,200,000 examples, and according to investigations, actual morbidity number than the high 5-8 of number of reports doubly.Suffering from the chronic hepatitis patient now is 1,200 ten thousand examples, die from 300,000 examples of hepatopathy every year, wherein the highest by the hepatitis gravis liver failure sickness rate due to the hepatitis B, account for that about 3,000 ten thousand people may become chronic hepatitis patient among the 1.2 hundred million hepatitis B virus infection persons of whole world HBV number of the infected 1/3, it more than 1% hepatitis gravis, add the hepatitis gravis that hepatitis viruss such as third type, fourth type are independent or superinfection takes place, the situation is tense.Though country has organized repeatedly tackling key problem, treatment does not still have good plan, and case fatality rate is still up to 60-80%.Viral hepatitis is the serious harm people ' s health not only, and brings heavy economic losses for country and individual, China every year due to illness the direct economic loss that causes of virus hepatitis at least about 300-500 hundred million RMB.For this disease, the treatment means of using roughly has following several at present:
1. the secular clinical experience of Drug therapy proves, for the liver failure patient, because liver has been lost regeneration and compensation, Drug therapy is difficult to fundamentally make the hepatic tissue restore funcitons, so can not obtain desirable therapeutic.
2. the treatment of abiotic type artificial liver support system, dialyse by blood/plasma, or adopt activated carbon and ion exchange resin to carry out the blood/plasma perfusion, or means such as plasmapheresis, remove toxicant and chemical substances such as aromatic amino acid, phenol in the blood, to alleviate the burden of liver, the expectation patient recovers self hepatocellular compensation gradually.Artificial hepaticsupport system commonly used at present generally is divided into blood absorbent-type and plasmapheresis type.Blood absorbent-type artificial liver is owing to the adsorbing material poor selectivity, and the histocompatibility of material own is relatively poor, also will remove human body beneficial's material absorption in the absorption noxious substance, causes the particularly hematoblastic minimizing of hemocyte.Plasmapheresis abandons after by plasma separator the separating plasma in the blood being gone out, give the patient with normal person's blood plasma infusion again, certain biotic component such as albumin, thrombin when removing toxicant, have also been replenished, but still abandoned in the blood plasma human body beneficial's material, consumed a large amount of normal persons' blood plasma simultaneously.
3. high activity external source hepatocyte is cultivated in the treatment of biological artificial liver support system in bioreactor, then patient's blood plasma is circulated by bioreactor, both carry out mass exchange by semipermeable membrane, utilize functions such as hepatocellular synthetic, detoxifcation of external source and biotransformation to provide support for the liver failure patient.Previously studies show that biological artificial liver support system can for liver decline the patient further treatment gain time.
Chinese patent 01115670.8 (publication number CN1383898A) discloses composite artificial liver supporting system and using method thereof.Comprise the toxicant removed in the blood/plasma and blood/plasma displacement (or absorption) device and Biotype artificial liver device of harmful substance.The former comprises: be used to separate patient's blood plasma and will separate after the plasma separating unit of blood plasma discharge; The healthy blood plasma gatherer that pump, valve and pipeline are arranged; And make above-mentioned blood/plasma displacement (absorption) device be in thermostat under the human body temperature.The latter comprises: the bioreactor of being made up of incubator and mass exchange device; The blood/plasma constant-current device; Culture fluid constant-current device and thermostat.Can support liver failure patient's liver function on this technical know-how at certain hour, but also exist not enough: one, lack gas supply, control device.The hepatocyte certain density CO of living necessities that exsomatizes
2And O
2, traditional hepatocellular condition of culture is 37 ℃ of following 5%CO
2With 95% air.Need a large amount of exogenous hepatocyte of High Density Cultivation of exsomatizing in Biotype artificial liver back-up system, the oxygen that hepatocyte density height thereby hepatocyte obtain is limited, should strengthen oxygen and be provided with keeping its vigor and function.Foreign study shows 5%CO
2At least 30%O
2Concentration helps hepatocyte existence most.Two, the aggregation that hepatocyte forms has influenced therapeutic effect.New isolating hepatocyte is difficult to keep its activity and function owing to lacking three-dimensional bracket in suspension, clinic trial only can change hepatic insufficiency patient's liver function the short time.Above-mentioned patent application discloses the characteristic of utilizing hepatocyte to have the gathering growth and has adopted the adherent method of restriction as vibration etc. new isolating hepatocyte to be assembled mutually, form many cells spheroplast, but spheroplast is not of uniform size, be deposited on the bottom easily, interior confluent monolayer cells can not get prescribing adequate nutrition and oxygen, the death that comes off easily of outer field cell, is stopped up the fiber pipe exocoel at easily stifled film when flowing through bioreactor, and culture fluid flows and the mass exchange all effected.
Summary of the invention:
The purpose of this utility model is to overcome existing biological artificial liver support system deficiency, and a kind of human liver's of replacement biological artificial liver support system is provided.Be specifically related to a kind of stirring microcarrier formula Biotype artificial liver back-up system.In more detail, provide that a kind of and mechanical artificial liver is used in combination, can substitute liver failure patient liver function in a long time, support the severe liver patient that declines to wait for liver transplantation, perhaps make the liver failure patient recover self hepatic tissue and its regeneration and compensation gradually, patient's survival rate and the biological artificial liver support system of time-to-live thereby the raising liver declines.
This utility model is by the bioreactor that can carry out mass exchange; Incubator with stir culture bottle function; Blood plasma/culture fluid constant-current device; Magnetic stirrer at a slow speed; Thermostat; Membrane oxygenator; Make in CO2 and the supply of O2 gas, control device and blood plasma pond.
This utility model provides at the deficiency of existing similar inventions: 1. O2 gas supply, control device; 2. incubator with stir culture bottle function is provided; 3. corresponding magnetic stirrer at a slow speed; 4. membrane oxygenator also is provided, help each hepatocyte and obtain sufficient oxygen supply and oxygenate, native system can be cultivated microcarrier formula training method and combine with stirring-type, growth provides bigger surface area to cell attachment, utilizes nutritional labeling and gas in the cultivating system fully.
Bioreactor of the present utility model comprises: 1, mass exchange device, its film surface area that can be used for mass exchange reaches 2m
2, wherein be provided with import (1a) and outlet (1b) that patient's blood plasma is come in and gone out; Be used to contain import (1c) and the outlet (1d) that external source liver cell culture liquid is come in and gone out; Be used to add fresh medium (1a) or the fresh import that contains external source liver cell culture liquid (1c); With the boundling fiber pipe that semipermeable membrane is made, it separates patient's blood plasma and external source liver cell culture liquid, in can carrying out between the two, low molecular mass exchange.2, one is used for introducing and cultivates active exogenous hepatocellular stir culture bottle: bottle is provided with one and contains liver cell culture liquid with above-mentioned bioreactor and export the import that contains exogenous liver cell culture liquid (2b) that is communicated with, one with above-mentioned bioreactor in contain the outlet that contains exogenous liver cell culture liquid (2a) that the import of liver cell culture liquid is communicated with; A permeability cell is arranged in the culture bottle, and ambient atmosphere can infiltrate through stir culture bottle (2c) by permeability cell; A revisal pipe (2d) is arranged in this culture bottle, can add, change culture fluid through this pipe; Also be equipped with a probe tube (2e) in the bottle, draw culture fluid and detect pH, PCO
2, PO
2Electrolyte etc., 3, make patient's blood plasma between above-mentioned bioreactor and human recycle system, carry out the circulation constant-current device, 4, make and contain exogenous hepatocellular culture fluid and between above-mentioned bioreactor and stir culture bottle, carry out circulation culture fluid constant-current device, 5, a magnetic stirrer at a slow speed, described magnetic stirrer at a slow speed can be simultaneously provides power at 1000 milliliters stir culture bottle at least for 4 capacity, can cultivate the 4000m1 hepatocyte simultaneously, 6, a membrane oxygenator that is used for to the oxygen supply of liver cell culture liquid: this oxygenator is provided with oxygen inlet (6a) and oxygen outlet (6b), be used to allow and contain the import (6c) of external source liver cell culture liquid discrepancy and export (6d), described oxygen inlet is connected with the oxygen supply pipe, oxygen supply pipe and the shared oxygen supply pipeline of native system oxygen supply device, oxygen outlet is opened in the case, 7, one is used for sterilizing ultraviolet lamps, described being arranged in the casing, 8, illuminating lamp, and a CO
2Feeding mechanism is guaranteed CO
2Concentration between 0-20%, an O
2Feeding mechanism is guaranteed O
2Concentration is between 0-95%, one is used to make this bioreactor to remain on thermostat (casing) under the human body temperature, a fan, guarantee that exogenous liver cell culture ambient temperature, gas concentration and humidity keep homogeneous, a control ejector is with constant-current device, magnetic stirrer, thermostat, CO at a slow speed
2Feeding mechanism, O
2The feeding mechanism centralized Control, a blood plasma pond, make do not disturb mutually between the human circulation regulating liver-QI cell culture fluid circulation with one be used for the filterable device of little bolt, prevent that cell debris class material from entering human circulation, described little bolt filtering device wherein filter membrane aperture is 0.22～0.45 micron.
This utility model needs and the abiotic artificial liver coupling with separating plasma function, also can share with the abiotic artificial liver of various physical property antidotal.This utility model has similar instrumental function now more comprehensively, and cell culture system is more perfect rationally, and gas supply of the present utility model and control device can provide high concentration, regulatable CO
2And O
2The stir culture system that is adopted adds micro-carrier system, cultivate exogenous hepatocyte, be more suitable for hepatic cell growth, especially the hepatocellular cultivation of high density, can not only obviously increase and cultivate hepatocellular quantity, and can keep the good morphological characteristic of hepatocyte, keep albumen synthetic and bilirubin metabolism function and function of detoxification by three-dimensional supporting role is provided.Can carry out the microcarrier cell culture by this utility model, and have the following advantages: (1) offers the cell attachment growth in unit volume surface area is big, and the ratio of growth area/volume is big, and the space of cells survival is abundant, cultured cell density height; (2) microcarrier is accurate suspended state, can make full use of nutritional labeling and gas in the cultivating system, the solid space of cell growing three-dimensional is provided, for the free cell that suspends is provided by adherent adequate time and the space of providing, during especially for the mixing liver cell culture, form the microstructure of similar liver structure between hepatocyte easily, help keeping of hepatocellular growth and function.Main is when containing the hepatocellular culture fluid of microcarrierization and flowing between stir culture bottle and bioreactor, and hydrodynamics is more stable, and hepatocyte injury is littler.The new blood plasma pond of while this utility model, the circulation of human circulation regulating liver-QI cell culture fluid is not disturbed mutually, neither influence blood samples of patients kinetics, can accelerate liver cell culture liquid circulation rate again with blood plasma that strengthens the patient and culture fluid carries out mass exchange by semipermeable membrane efficient.
Mechanical part that this utility model adopts is commercial.
Description of drawings
Fig. 1 is this utility model structural representation.
Wherein: 1, mass exchange device, 2, the stir culture bottle, 3, the circulation constant-current device, 4, the culture fluid constant-current device, 5, magnetic stirrer at a slow speed, 6, membrane oxygenator, 7, Burdick lamp, 8, illuminating lamp.Fig. 2 is this utility model schematic appearance.
Wherein: 1, side opening 1,2, side opening 2,3, side opening 3.
The specific embodiment
Further specifying this utility model in conjunction with following embodiment connects and implementation method.
Hepatocyte is flowed (containing the blood plasma pond) at the hollow fiber conduit exocoel during treatment, use abiotic artificial liver displacement patient blood plasma earlier, remove most of toxicant in patient's blood plasma, the blood plasma pond is stayed in the storage of patient's part blood plasma, blood plasma constant-current device (3) in the Biotype artificial liver apparatus for supporting of the present invention then, by blood plasma being introduced in the Biotype artificial liver apparatus for supporting through side opening 3, again through above-mentioned bioreactor blood plasma import (1a), the blood plasma in the blood plasma pond imported in the above-mentioned bioreactor (1) carry out mass exchange; Simultaneously, Biotype artificial rami hepatici of the present invention holds that culture fluid constant-current device (4) will contain the exogenous hepatocellular culture fluid of the fresh microcarrierization of q.s in the system in the stir culture bottle (2), through side arm (2a) input oxygenator (6c), this moment, oxygen was entered by oxygen inlet (6a), (6b) disengages by oxygen outlet, and flow out the culture fluid import (1c) by above-mentioned bioreactor again of hepatocellular culture fluid from oxygenator (6d), in the input bioreactor (1), in making that above-mentioned patient's blood plasma and hollow fiber conduit that above-mentioned culture fluid is made by semipermeable membrane carry out, the small-molecule substance exchange, finish synthetic, metabolism, liver functions such as biotransformation, then of the culture fluid outlet (1d) of the culture fluid of having finished liver mass exchange function by bioreactor, be transported in the stir culture bottle (2b) with culture fluid constant-current device (4), blood plasma through the patient after the mass exchange then exports (1b) by the blood plasma in the bioreactor, send it back to blood plasma pond with blood plasma constant-current device (3) through side opening 2, again in the blood circulation by the defeated the Huis' body of abiotic artificial liver.Used connecting line is medical silicone tube or medical poly-third ethylene tube.Biotype artificial liver apparatus for supporting side opening 1 is drawn the outlet of pipeline for the permeability cell of stir culture device, revisal pipe and probe tube.
Microcarrier with the fresh medium dissolving disinfects will contain the sterilization stir culture bottle (2) that the microcarrier culture fluid injects silication again, place 37 ℃ to contain 5%CO in the stir culture bottle
2More than the 30min, then isolating exogenous hepatocyte is pressed 1 * 10 among the bioartificial liver of high-concentration oxygen
7The ratio of/ml is seeded in the blender jar, slowly stirs, and changes 1/2 culture medium every other day.
Cultivate in the embodiment 3 hepatocyte doughnuts
The exogenous hepatocyte that separates the back microcarrierization can be through above-mentioned bioreactor culture liquid import (1c), in the input bioreactor (1), make hepatocyte be evenly distributed on bioreactor hollow fiber conduit exocoel, culture fluid constant-current device (3) is drawn culture fluid from stir culture bottle (2a) then, input oxygenator (6c), enter bioreactor doughnut tube cavity by above-mentioned bioreactor blood plasma import (1b) again, hepatocyte is by nutritional labeling and oxygen in the semipermeable membrane picked-up culture fluid, discharge metabolite, culture fluid constant-current device (4) is failed culture fluid and is got back to stir culture bottle (2b) by the outlet of the blood plasma in the bioreactor (1a).Hepatocyte was fixed in the hollow fiber conduit exocoel when the exogenous hepatocyte that this method is cultivated was used for the treatment of.
Hepatocyte is flowed (containing the blood plasma pond) at the hollow fiber conduit exocoel during embodiment 4 treatments
Use abiotic artificial liver displacement patient blood plasma earlier, remove most of toxicant in patient's blood plasma, the blood plasma pond is stayed in the storage of patient's part blood plasma, blood plasma constant-current device (3) in the Biotype artificial liver of the present invention then system, by above-mentioned bioreactor blood plasma import (1a), the blood plasma in the blood plasma pond imported carry out mass exchange in the above-mentioned bioreactor; Simultaneously, Biotype artificial rami hepatici of the present invention holds that the culture fluid constant-current device will contain the exogenous hepatocellular culture fluid of the fresh microcarrierization of q.s in the system in the culture fluid, input oxygenator (6c) is the culture fluid import (1c) by above-mentioned bioreactor again, in the input bioreactor (1), in making that above-mentioned patient's blood plasma and hollow fiber conduit that above-mentioned culture fluid is made by semipermeable membrane carry out, the small-molecule substance exchange, finish synthetic, metabolism, liver functions such as biotransformation, then of the culture fluid outlet (1d) of the culture fluid of having finished liver mass exchange function by bioreactor, be transported in the stir culture bottle (2b) with the culture fluid constant-current device, blood plasma through the patient after the mass exchange then exports (1b) by the blood plasma in the bioreactor, send it back to blood plasma pond with the blood plasma constant-current device, again in the blood circulation by the defeated the Huis' body of abiotic artificial liver.
Hepatocyte is at hollow fiber conduit exocoel mobile (depletion of blood stock tank) when embodiment 5, treatment
Use abiotic artificial liver displacement patient blood plasma earlier, remove most of toxicant in patient's blood plasma, blood plasma constant-current device (3) in the Biotype artificial liver of the present invention then system, by above-mentioned bioreactor blood plasma import (1a), carry out mass exchange importing in the above-mentioned bioreactor (1) through the blood plasma after the displacement; Simultaneously, Biotype artificial rami hepatici of the present invention holds that culture fluid constant-current device (4) will contain the exogenous hepatocellular culture fluid of the fresh microcarrierization of q.s in the system in the culture fluid, input oxygenator (6c) is the culture fluid import (1c) by above-mentioned bioreactor again, in the input bioreactor (1), in making that above-mentioned patient's blood plasma and hollow fiber conduit that above-mentioned culture fluid is made by semipermeable membrane carry out, the small-molecule substance exchange, finish synthetic, metabolism, liver functions such as biotransformation, then of the culture fluid outlet (2b) of the culture fluid of having finished the liver function mass exchange by bioreactor, be transported in the stir culture bottle (2) with culture fluid constant-current device (4), blood plasma through the patient after the mass exchange then exports (1b) by the blood plasma in the bioreactor, with blood plasma constant-current device (3) it is sent back in the blood circulation of human body.
Hepatocyte is fixed in hollow fiber conduit exocoel (containing the blood plasma pond) during embodiment 7 treatments
Use abiotic artificial liver displacement patient blood plasma earlier, remove most of toxicant in patient's blood plasma, the blood plasma pond is stayed in the storage of patient's part blood plasma, blood plasma constant-current device (3) in the Biotype artificial liver of the present invention then system, by above-mentioned bioreactor blood plasma import (1a), blood plasma in the blood plasma pond imported in the above-mentioned bioreactor (1) carry out mass exchange, in making that above-mentioned patient's blood plasma and hollow fiber conduit that above-mentioned exogenous liver cell culture liquid is made by semipermeable membrane carry out, the small-molecule substance exchange, finish synthetic, metabolism, liver functions such as biotransformation, then the blood plasma through the patient after the mass exchange is then exported (1b) by the blood plasma in the bioreactor, send it back to blood plasma pond with blood plasma constant-current device (3), again in the blood circulation by the defeated the Huis' body of abiotic artificial liver.
Hepatocyte is fixed in hollow fiber conduit exocoel (depletion of blood stock tank) during embodiment 8 treatments
Use abiotic artificial liver displacement patient blood plasma earlier, remove most of toxicant in patient's blood plasma, blood plasma constant-current device (3) in the Biotype artificial liver of the present invention then system, by above-mentioned bioreactor blood plasma import (1a), carry out mass exchange importing in the above-mentioned bioreactor (1) through the blood plasma after the displacement, in making that above-mentioned patient's blood plasma and hollow fiber conduit that above-mentioned exogenous liver cell culture liquid is made by semipermeable membrane carry out, the small-molecule substance exchange, finish synthetic, metabolism, liver functions such as biotransformation, then the blood plasma through the patient after the mass exchange is then exported (1b) by the blood plasma in the bioreactor, it is sent back in the blood circulation of human body with blood plasma constant-current device (3).
Claims (7)
1, a kind of stirring microcarrier formula Bioartificial Liver Support System, comprise casing and control circuit, bioreactor, the thermostat that carry out carrying out circulation culture fluid constant-current device between circulation constant-current device, bioreactor and the stir culture bottle between bioreactor and the human recycle system, keeps the biological respinse actuator temperature, it is characterized in that also comprising incubator with stir culture bottle function, magnetic stirrer at a slow speed, membrane oxygenator, CO
2And O
2Gas supply, control device and blood plasma pond and filter the device of little bolt, described its film surface area of bioreactor that carries out mass exchange reaches 2m
2
2, stirring microcarrier formula Bioartificial Liver Support System according to claim 1 is characterized in that described constant-current device, magnetic stirrer, thermostat, CO2 feeding mechanism, O2 feeding mechanism centralized Control at a slow speed.
3, biological artificial liver support system according to claim 1 is characterized in that, the thermostat among the described stirring microcarrier formula Bioartificial Liver Support System is the small-sized temperature automatically controlled air-conditioner of energy.
4, biological artificial liver support system according to claim 1 is characterized in that the oxygen inlet of described membrane oxygenator device is connected with the oxygen supply pipe, oxygen supply pipe and the shared oxygen supply pipeline of native system oxygen supply device.
5, biological artificial liver support system as claimed in claim 1, it is characterized in that described little bolt filtering device wherein the filter membrane aperture be 0.22～0.45 micron.
6, biological artificial liver support system according to claim 1 is characterized in that in the described casing Burdick lamp being set.
7, stirring microcarrier formula Bioartificial Liver Support System according to claim 1 is characterized in that described magnetic stirrer at a slow speed stirs 4 high capacity culture bottles simultaneously.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200320109050 CN2661199Y (en) | 2003-10-21 | 2003-10-21 | Microcarrier stirred type biological artificial liver support system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200320109050 CN2661199Y (en) | 2003-10-21 | 2003-10-21 | Microcarrier stirred type biological artificial liver support system |
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| Publication Number | Publication Date |
|---|---|
| CN2661199Y true CN2661199Y (en) | 2004-12-08 |
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ID=34342979
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200320109050 Expired - Fee Related CN2661199Y (en) | 2003-10-21 | 2003-10-21 | Microcarrier stirred type biological artificial liver support system |
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| CN (1) | CN2661199Y (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102725008A (en) * | 2009-08-07 | 2012-10-10 | 汉莫堤克股份有限公司 | Device and method for eliminating biologically harmful substances from bodily fluids |
| CN106139289A (en) * | 2016-07-29 | 2016-11-23 | 武汉仝干医疗科技股份有限公司 | Semipermeable membrane layering integrated biological reactor |
-
2003
- 2003-10-21 CN CN 200320109050 patent/CN2661199Y/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102725008A (en) * | 2009-08-07 | 2012-10-10 | 汉莫堤克股份有限公司 | Device and method for eliminating biologically harmful substances from bodily fluids |
| CN106139289A (en) * | 2016-07-29 | 2016-11-23 | 武汉仝干医疗科技股份有限公司 | Semipermeable membrane layering integrated biological reactor |
| CN106139289B (en) * | 2016-07-29 | 2018-06-19 | 武汉仝干医疗科技股份有限公司 | Semi-permeable membrane is layered integrated biological reactor |
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| CF01 | Termination of patent right due to non-payment of annual fee |