CN111383774A - System for screening treatment regimens for brain gliomas - Google Patents
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Abstract
The present disclosure relates to a system for screening brain glioma treatment protocols, the system comprising a culture device, an input device, a computing device, and an output device; wherein, the culture device is used for culturing to obtain brain glioma organoids; the input device is used for inputting the tumor cell proliferation level, the immune cell positive rate and the angiogenesis level of the brain glioma organoid to be detected; wherein the level of tumor cell proliferation comprises a Ki67 positivity rate; the multiple immune cell positive rates comprise a microglia positive rate, a cytotoxic T cell positive rate, a helper T cell positive rate and a natural killer cell positive rate; the angiogenesis levels comprise the positive rate of the blood vessel marker molecule CD31 and the positive rate of CD 105; the output device is used for outputting the effectiveness of the brain glioma treatment scheme. The screening result of the system for the treatment scheme of the glioma can truly and accurately judge the treatment effect of the treatment scheme of the glioma on the glioma tissues in the body of the patient.
Description
Technical Field
The disclosure relates to the technical field of biological medicine, in particular to a system for screening a treatment scheme of brain glioma.
Background
The brain glioma is the primary intracranial tumor with the highest incidence rate, accounts for about 46 percent of the intracranial tumor, and accounts for about 31.1 percent of the intracranial tumor in the middle-aged and young people aged 20-59 in China. In order to better guide the clinical treatment of patients with brain glioma, a reasonable treatment scheme for brain glioma needs to be screened out.
In the related art, a glioma cell line is used as an in vitro glioma model for screening glioma treatment protocols. However, the screening results of the glioma treatment schemes using the glioma cell lines are still not accurate enough, and the therapeutic effects of the glioma treatment schemes on glioma tissues cannot be truly reflected.
Disclosure of Invention
The purpose of the present disclosure is to provide a system for screening a glioma treatment plan, which is used to solve the problem that the screening result of the existing screening glioma treatment plan cannot truly reflect the treatment effect of the glioma treatment plan on glioma tissues.
In order to achieve the above object, the present disclosure provides a system for screening a brain glioma treatment plan, characterized in that the system comprises a culturing device, an input device, a computing device and an output device; wherein the content of the first and second substances,
the culture device is used for realizing the following culture operation steps:
a. mixing the brain glioma tissue block with a culture medium, and culturing to obtain a brain glioma organoid;
b. according to a preset treatment scheme of the brain glioma, treating and culturing the brain glioma organoid to obtain a brain glioma organoid to be detected;
c. obtaining the proliferation level of tumor cells, the positive rates of a plurality of immune cells and the angiogenesis level of the brain glioma organoid to be detected;
the input device is used for inputting the tumor cell proliferation level, the immune cell positive rate and the angiogenesis level of the brain glioma organoid to be detected; wherein the level of tumor cell proliferation comprises a Ki67 positivity rate; the multiple immune cell positive rates comprise a microglia positive rate, a cytotoxic T cell positive rate, a helper T cell positive rate and a natural killer cell positive rate; the angiogenesis levels comprise the positive rate of the blood vessel marker molecule CD31 and the positive rate of CD 105;
the computing device comprises a memory having a computer program stored therein and a processor configured to execute the computer program stored in the memory to implement a modeling algorithm and an algorithm of a discriminant function as shown in equation (1); the modeling algorithm is a support vector machine algorithm and/or a minimum deviation two-times algorithm;
F(c)＝sgn[f1(c1)+f2(c2)+f3(c3)+f4(c4)+f5(c5)+f6(c6)+f7(c7)+b]
formula (1)
In the formula (1), F (c) represents the effectiveness of the treatment scheme of the brain glioma, F (c) returns that the value is-1 to represent the effectiveness, returns that the value is 1 to represent the ineffectiveness; c. C1、c2、c3、c4、c5、c6And c7Respectively and sequentially showing Ki67 positive rate, microglia positive rate, cytotoxic T cell positive rate, helper T cell positive rate, natural killer cell positive rate, CD31 positive rate and CD105 positive rate; f. of1(c1)、f2(c2)、f3(c3)、f4(c4)、f5(c5)、f6(c6) And f7(c7) Respectively obtaining kernel functions obtained by training according to the modeling algorithm, and b obtaining critical score values obtained by training according to the modeling algorithm;
the output device is used for outputting the effectiveness of the brain glioma treatment scheme.
Optionally, the culture medium in step a comprises DMEM/F12 medium, Neurobasal medium, cell culture additive Glutamax, non-essential amino acids NEAAs, vitamin a-free cell culture additive B-27, penicillin-streptomycin, and insulin.
Optionally, in the culture medium, the DMEM/F12 culture medium is used in an amount of 0.4-0.6 parts by volume, the Neurobasal medium Neurobasal is used in an amount of 0.4-0.6 parts by volume, the cell culture additive Glutamax is used in an amount of 0.005-0.02 parts by volume, the non-essential amino acids NEAAs are used in an amount of 0.005-0.02 parts by volume, the cell culture additive B-27 containing no vitamin a is used in an amount of 0.01-0.04 parts by volume, the penicillin-streptomycin is used in an amount of 0.005-0.02 parts by volume, and the insulin is used in an amount of 1.5-4 micrograms per milliliter.
Optionally, the glioma tissue mass has a diameter of 0.3-1 cm.
Optionally, when the glioma tissue block is mixed with the culture medium in step a, the amount of the culture medium is 1-10mL relative to 100mg of the glioma tissue block.
Optionally, the culturing in step a results in a brain glioma organoid comprising:
placing the mixture of the brain glioma tissue block and the culture medium in an incubator for shake culture, replacing the culture medium every 2-3 days in the culture process, and continuously culturing for 3-21 days to obtain the brain glioma organoid; wherein the culture conditions comprise: the temperature is 34-40 ℃, and the concentration of carbon dioxide is 4-8%; the shaking speed of the shaking culture is 70-150 rpm.
Alternatively, in the formula (1), f1(c1)＝0.7×c1，f2(c2)＝0.05×c2，f3(c3)＝0.05×c33，f4(c4)＝0.05×c4，f5(c5)＝0.05×c5，f6(c6)＝0.05×c6，f7(c7)＝0.05×c7，b＝-0.15。
Optionally, the microglial cell positive rate comprises a microglial marker molecule Ibal positive rate, the cytotoxic T cell positive rate comprises a cytotoxic T cell marker molecule CD8 positive rate, and the helper T cell positive rate comprises a helper T cell marker molecule CD4 positive rate; the natural killer cell positive rate comprises the positive rate of the marker molecule CD56 of the natural killer cells.
The present disclosure also provides a molecular marker for screening a treatment regimen for brain glioma, the molecular marker comprising a marker molecule Ki67 for tumor cell proliferation level, a marker molecule Ibal for microglia, a marker molecule CD8 for cytotoxic T cells, a marker molecule CD4 for helper T cells, a marker molecule CD56 for natural killer cells, a marker molecule CD31 for angiogenesis, and a marker molecule CD105 for angiogenesis.
The present disclosure also provides use of an agent for detecting molecular markers including a marker molecule Ki67 for tumor cell proliferation level, a marker molecule Iba1 for microglia, a marker molecule CD8 for cytotoxic T cells, a marker molecule CD4 for helper T cells, a marker molecule CD56 for natural killer cells, a marker molecule CD31 for angiogenesis, and a marker molecule CD105 for angiogenesis in the preparation of a kit for screening a treatment regimen for brain glioma; the reagent for detecting the molecular marker can detect the positive rate of the molecular marker.
According to the technical scheme, the system for screening the treatment scheme of the brain glioma, which is provided by the disclosure, is provided with the culture device, and can be used for culturing the brain glioma organoids and screening the brain glioma organoids obtained by culture for the treatment scheme of the brain glioma; meanwhile, the discrimination algorithm provided by the disclosure can accurately and quickly discriminate the treatment effect of the brain glioma treatment scheme.
Additional features and advantages of the disclosure will be set forth in the detailed description which follows.
Detailed Description
The following describes in detail specific embodiments of the present disclosure. It should be understood that the detailed description and specific examples, while indicating the present disclosure, are given by way of illustration and explanation only, not limitation.
A first aspect of the present disclosure provides a system for screening a glioma treatment protocol, the system comprising a culture device, an input device, a computing device, and an output device; wherein the culture device is used for realizing the following culture operation steps: a. mixing the brain glioma tissue block with a culture medium, and culturing to obtain a brain glioma organoid; b. according to a preset treatment scheme of the brain glioma, treating and culturing the brain glioma organoid to obtain a brain glioma organoid to be detected; c. obtaining the proliferation level of tumor cells, the positive rates of a plurality of immune cells and the angiogenesis level of the brain glioma organoid to be detected; the input device is used for inputting the tumor cell proliferation level, the immune cell positive rate and the angiogenesis level of the brain glioma organoid to be detected; wherein the level of tumor cell proliferation comprises a Ki67 positivity rate; the multiple immune cell positive rates comprise a microglia positive rate, a cytotoxic T cell positive rate, a helper T cell positive rate and a natural killer cell positive rate; the angiogenesis levels comprise the positive rate of the blood vessel marker molecule CD31 and the positive rate of CD 105; the computing device comprises a memory having a computer program stored therein and a processor configured to execute the computer program stored in the memory to implement a modeling algorithm and an algorithm of a discriminant function as shown in equation (1); the modeling algorithm is a support vector machine algorithm and/or a minimum deviation two-times algorithm;
F(c)＝sgn[f1(c1)+f2(c2)+f3(c3)+f4(c4)+f5(c5)+f6(c6)+f7(c7)+b]
formula (1)
In the formula (1), F (c) represents the effectiveness of the treatment scheme of the brain glioma, F (c) returns that the value is-1 to represent the effectiveness, returns that the value is 1 to represent the ineffectiveness; c. C1、c2、c3、c4、c5、c6And c7Respectively and sequentially showing Ki67 positive rate, microglia positive rate, cytotoxic T cell positive rate, helper T cell positive rate, natural killer cell positive rate, CD31 positive rate and CD105 positive rate; f. of1(c1)、f2(c2)、f3(c3)、f4(c4)、f5(c5)、f6(c6) And f7(c7) Respectively, the kernels obtained by training according to the modeling algorithmB is a critical score value obtained by training according to the modeling algorithm; the output device is used for outputting the effectiveness of the brain glioma treatment scheme.
Through the technical scheme, the system for screening the treatment scheme of the brain glioma, which is provided by the disclosure, is provided with the culture device, the brain glioma organoid can be cultured, and the brain glioma organoid obtained by culture is used for screening the treatment scheme of the brain glioma.
The technical scheme of the disclosure provides a high-specificity and high-sensitivity biomarker and a curative effect prediction model for the curative effect screening of a glioma treatment scheme, and provides a novel and efficient auxiliary tool for clinically formulating the glioma treatment scheme.
According to the present disclosure, in order to increase the success rate of culturing the brain glioma organoids, the culture medium in step a preferably comprises DMEM/F12 culture medium, Neurobasal medium, cell culture additive Glutamax, non-essential amino acids NEAAs, cell culture additive B-27 without vitamin a, penicillin-streptomycin and insulin. The culture medium containing the components can provide proper nutrient substances for growth and proliferation of the glioma cells and cells in a glioma microenvironment, provide basic conditions for directly utilizing the glioma tissue blocks to culture glioma organoids, and improve the success rate of glioma organoid culture.
According to the present disclosure, the amount of each component in the culture medium may vary within a wide range, for example, the DMEM/F12 culture medium may be used in an amount of 0.4 to 0.6 parts by volume, the Neurobasal medium may be used in an amount of 0.4 to 0.6 parts by volume, the cell culture additive Glutamax may be used in an amount of 0.005 to 0.02 parts by volume, the non-essential amino acids NEAAs may be used in an amount of 0.005 to 0.02 parts by volume, the cell culture additive B-27 containing no vitamin a may be used in an amount of 0.01 to 0.04 parts by volume, the penicillin-streptomycin may be used in an amount of 0.005 to 0.02 parts by volume, and the insulin may be used in an amount of 1.5 to 4 micrograms per milliliter. The components are all commercial products, and details of specific components and contents of the components are not repeated in the disclosure.
Preferably, the brain glioma bulk has a diameter of 0.3-1 cm. The brain glioma tissue block is directly used for culturing the brain glioma organoid, and the cultured brain glioma organoid can simulate the brain glioma tissue microenvironment in a patient body more truly, so that the reliability of the detection result is further improved.
Preferably, the step a further comprises the step of shearing the brain glioma tissue at a low temperature to obtain the brain glioma tissue mass, and then performing the operation of the step a.
According to the present disclosure, the relative amount of the glioma tissue mass to the culture medium when the glioma tissue mass is mixed with the culture medium in step a can vary within a wide range, for example, the amount of the culture medium is 1-10mL per 100mg of the glioma tissue mass when the glioma tissue mass is mixed with the culture medium in step a.
Optionally, the culturing in step a results in a brain glioma organoid comprising: placing the mixture of the brain glioma tissue block and the culture medium in an incubator for shake culture, replacing the culture medium every 2-3 days in the culture process, and continuously culturing for 3-21 days to obtain the brain glioma organoid; wherein the culture conditions comprise: the temperature is 34-40 ℃, and the concentration of carbon dioxide is 4-8%; the shaking speed of the shaking culture is 70-150 rpm.
Alternatively, in the formula (1), f1(c1)＝0.7×c1，f2(c2)＝0.05×c2，f3(c3)＝0.05×c33，f4(c4)＝0.05×c4，f5(c5)＝0.05×c5，f6(c6)＝0.05×c6，f7(c7)＝0.05×c7And b is-0.15. In addition, f is1(c1)、f2(c2)、f3(c3)、f4(c4)、f5(c5)、f6(c6)、f7(c7) And b may vary with the bias of the marker molecule selection and detection means, or may vary with factors such as the size of the data scale of the training data set. F above1(c1)、f2(c2)、f3(c3)、f4(c4)、f5(c5)、f6(c6)、f7(c7) And b is obtained by training the data in the embodiment with a modeling algorithm of a minimum-deviation two-times, and does not limit the scope of the disclosure.
Preferably, the microglial cell positive rate comprises a microglial marker molecule Ibal positive rate, the cytotoxic T cell positive rate comprises a cytotoxic T cell marker molecule CD8 positive rate, and the helper T cell positive rate comprises a helper T cell marker molecule CD4 positive rate; the natural killer cell positive rate comprises the positive rate of the marker molecule CD56 of the natural killer cells.
A second aspect of the present disclosure provides a molecular marker for screening a treatment regimen for brain glioma, wherein the molecular marker includes a marker molecule Ki67 for a proliferation level of tumor cells, a marker molecule Ibal for microglia, a marker molecule CD8 for cytotoxic T cells, a marker molecule CD4 for helper T cells, a marker molecule CD56 for natural killer cells, a marker molecule CD31 for angiogenesis, and a marker molecule CD105 for angiogenesis.
A third aspect of the present disclosure provides a use of an agent for detecting molecular markers including a marker molecule Ki67 at a tumor cell proliferation level, a marker molecule Ibal of microglia, a marker molecule CD8 of cytotoxic T cells, a marker molecule CD4 of helper T cells, a marker molecule CD56 of natural killer cells, a marker molecule CD31 of angiogenesis, and a marker molecule CD105 of angiogenesis in the preparation of a kit for screening a treatment regimen for brain glioma; the reagent for detecting the molecular marker can detect the positive rate of the molecular marker.
The present disclosure is further illustrated by the following examples, but is not to be construed as being limited thereby. The raw materials, reagents, instruments and equipment related to the embodiments of the present disclosure can be purchased from the market unless otherwise specified.
Where specific experimental temperatures are not noted in the examples of the present disclosure, the experimental temperatures are all room temperature (20-25 ℃). The sources of reagents used in the examples of the disclosure are as follows:
DMEM/F12 medium was purchased from Thermo Fisher Scientific, Neurobasal medium from Thermo Fisher Scientific, cell culture supplement Glutamax from Thermo Fisher Scientific, non-essential amino acids NEAAs from Thermo Fisher Scientific, cell culture supplement B-27 without vitamin A from Thermo Fisher Scientific, penicillin-streptomycin from Thermo Fisher Scientific, and insulin from ChemMedExpress.
The culture medium used in the embodiments of the present disclosure is in an autonomous configuration, comprising: 0.5 parts by volume of DMEM/F12 medium, 0.4 parts by volume of Neurobasal medium, 0.02 parts by volume of cell culture additive Glutamax, 0.02 parts by volume of non-essential amino acids NEAAs, 0.04 parts by volume of cell culture additive B-27 without vitamin a, 0.02 parts by volume of penicillin-streptomycin, 1.5 micrograms per milliliter of insulin.
The present disclosure is described below in examples with reference to brain gliomas.
Example 1
This example illustrates the culture of glioma organoids of the present disclosure
Collecting postoperative fresh brain glioma tissue, transporting to a laboratory on ice within 0.5 h, pre-cleaning with normal saline, then repeatedly cleaning twice with DMEM/F12 culture medium containing 1 × Normocin antibiotics to obtain cleaned brain glioma tissue, shearing the cleaned brain glioma tissue on ice into brain glioma tissue blocks with the diameter of 0.3-1cm, and cutting the brain glioma tissue blocks into brain glioma tissue blocks with the diameter of 0.3-1cmMixing the sheared brain glioma tissue block with a culture medium, and placing the mixture in a 6-well plate at 37 ℃ and 5% CO2The cell culture box is used for culturing, wherein each culture hole contains 3mL of culture medium and 100mg of glioma tissue block, a shaking table is used for continuously shaking at the speed of 100rpm in the culture process, the culture medium is replaced every 3 days, and the glioma organoid is continuously cultured for 18 days, so that the glioma organoid is obtained.
Example 2
This example illustrates the establishment of a model for screening a glioma treatment protocol according to the present disclosure
1. The treatment scheme comprises the following steps: the glioma organoids of example 1 were divided into 2 equal parts (meaning that after the organoid tissue pieces were mixed with the culture medium uniformly, they were equally divided into two equal parts by volume), and the glioma organoids of each part were photographed under a microscope and the diameter of each organoid tissue was recorded. And adding an anti-glioma drug (crizotinib) into one part of the culture system to ensure that the concentration of the crizotinib in the culture system is 3 mu mol/L, and then continuously culturing for 1 week by using the culture method in the embodiment 1 to obtain the glioma organoids of the group to be detected. The other of the 2 equal parts was added with the same volume of DMSO as the above crizotinib, and then cultured for another 1 week in the culture method of example 1 to obtain a control group (reference group) brain glioma organoid. Each organoid in the test group and the control group was photographed under a microscope, and the average diameters of the two groups of organoid tissues were counted, respectively.
Brain glioma organoids from 50 patients were cultured repeatedly as described above.
2. And (3) judging the curative effect: and when the average diameter of the tumor organoids of the group to be detected is less than or equal to 0.8 time of the average diameter of the tumor organoids of the control group, judging that the treatment scheme is effective, otherwise, judging that the treatment scheme is ineffective.
3. The judgment shows that 13 cases of the glioma organoids to be tested have the treatment effect, and 37 cases of the glioma organoids to be tested have no obvious treatment effect.
Taking the brain glioma organoid to be detected with the treatment effect and the brain glioma organoid without the treatment effect, respectively carrying out frozen section and/or paraffin embedding section to obtain brain glioma organoid section, and carrying out the following detection:
a. brain glioma cell proliferation level detection
Respectively staining the sections with Ki67, and counting the Ki67 positive rate;
b. positive rate of immune cells in brain glioma organoids
Taking the slices to respectively detect the positive rates of microglia, cytotoxic T cells, helper T cells and natural killer cells;
c. detection of concentration levels of brain glioma cell vascular markers
The sections were further stained with vascular markers CD31 and CD105, respectively, and tested for CD 31-positive rate and CD 105-positive rate.
And using the detection result of the brain glioma organoid to be detected with the treatment effect as a training set, using the detection result of the brain glioma organoid to be detected without the treatment effect as a test set, and adopting a partial least square method to construct a screening model of a brain glioma treatment scheme, wherein the model is as shown in formula (1):
F(c)＝sgn[f1(c1)+f2(c2)+f3(c3)+f4(c4)+f5(c5)+f6(c6)+f7(c7)+b]
formula (1)
In the formula (1), F (c) represents the effectiveness of the treatment scheme of the brain glioma, F (c) returns that the value is-1 to represent the effectiveness, returns that the value is 1 to represent the ineffectiveness; c. C1、c2、c3、c4、c5、c6And c7Respectively and sequentially showing Ki67 positive rate, microglia positive rate, cytotoxic T cell positive rate, helper T cell positive rate, natural killer cell positive rate, CD31 positive rate and CD105 positive rate; f. of1(c1)、f2(c2)、f3(c3)、f4(c4)、f5(c5)、f6(c6) And f7(c7) Respectively a kernel function obtained by training according to the modeling algorithm, b is obtained by training according to the modeling algorithmA threshold score value to; specifically, in the formula (1), f1(c1)＝0.7×c1，f2(c2)＝0.05×c2，f3(c3)＝0.05×c3，f4(c4)＝0.05×c4，f5(c5)＝0.05×c5，f6(c6)＝0.05×c6，f7(c7)＝0.05×c7And b is-0.15, that is to say the discriminant function is specifically:
F(c)＝sgn[0.7×c1+0.05×c2+0.05×c3+0.05×c4+0.05×c5+0.05
×c6+0.05×c7-0.15]
formula (2).
Example 3
This example illustrates the validation of a glioma treatment protocol screening model of the present disclosure
Re-culturing 50 groups of glioma organoids by using the method in example 1, performing treatment culture by using the treatment scheme in example 2 to obtain the glioma organoids to be tested, and classifying the cultured glioma organoids to be tested into an effective treatment group and an ineffective treatment group according to the discrimination method in example 2. Respectively detecting the Ki67 positive rate, the microglia marker molecule Ibal positive rate, the cytotoxic T cell marker molecule CD8 positive rate, the helper T cell marker molecule CD4 positive rate, the natural killer cell marker molecule CD56 positive rate, the angiogenesis marker molecule CD31 positive rate and the angiogenesis marker molecule CD105 positive rate of the brain glioma organ organs to be detected in the effective treatment group and the ineffective treatment group, further verifying the performance of the screening model of the brain glioma treatment scheme, and evaluating indexes including area under the operating characteristic curve (ROC curve for short) of a receiver (accurate under the curve, AUC) of prediction Accuracy (ACC). The result of the ACC analysis of the formula (2) was found to be 80% and the AUC to be 0.85.
Example 4
This example serves to illustrate the validation of the glioma treatment protocol screening model of the present disclosure.
The 3 groups of glioma organoids were re-cultured using the method of example 1, the glioma organoids were photographed under a microscope, and the diameter of each organoid tissue was recorded. And (3) performing treatment culture by using the treatment scheme of the embodiment 2 to obtain the brain glioma organoids to be detected, and dividing the brain glioma organoids to be detected into an effective treatment group and an ineffective treatment group by using the method and the discrimination model disclosed by the disclosure. Each organoid in the effective treatment group and the ineffective treatment group was photographed under a microscope, and the average diameters of the two groups of organoid tissues were counted.
The performance of the brain glioma treatment scheme screening model is further verified by a method of counting the diameters of each organoid tissue in the effective treatment group and the ineffective treatment group (the diameters are used as volume indexes because the brain glioma organoids are mainly spherical and spheroids in shape). The mean diameters of the effective treatment group and the ineffective treatment group are shown in table 1:
table 1:
as can be seen from table 1, the discrimination result of the discrimination model provided by the present disclosure on the treatment effect of the glioma treatment plan coincides with the discrimination result of the conventional discrimination method, and it can be seen that the discrimination model of the present disclosure can accurately and rapidly discriminate the treatment effect of the glioma treatment plan.
The preferred embodiments of the present disclosure have been described in detail above, however, the present disclosure is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present disclosure within the technical idea of the present disclosure, and these simple modifications all fall within the protection scope of the present disclosure.
It should be noted that, in the foregoing embodiments, various features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various combinations that are possible in the present disclosure are not described again.
In addition, any combination of various embodiments of the present disclosure may be made, and the same should be considered as the disclosure of the present disclosure, as long as it does not depart from the spirit of the present disclosure.
Claims (10)
1. A system for screening brain glioma treatment protocols, the system comprising a culture device, an input device, a computing device, and an output device; wherein the content of the first and second substances,
the culture device is used for realizing the following culture operation steps:
a. mixing the brain glioma tissue block with a culture medium, and culturing to obtain a brain glioma organoid;
b. according to a preset treatment scheme of the brain glioma, treating and culturing the brain glioma organoid to obtain a brain glioma organoid to be detected;
c. obtaining the proliferation level of tumor cells, the positive rates of a plurality of immune cells and the angiogenesis level of the brain glioma organoid to be detected;
the input device is used for inputting the tumor cell proliferation level, the immune cell positive rate and the angiogenesis level of the brain glioma organoid to be detected; wherein the level of tumor cell proliferation comprises a Ki67 positivity rate; the multiple immune cell positive rates comprise a microglia positive rate, a cytotoxic T cell positive rate, a helper T cell positive rate and a natural killer cell positive rate; the angiogenesis levels comprise the positive rate of the blood vessel marker molecule CD31 and the positive rate of CD 105;
the computing device comprises a memory having a computer program stored therein and a processor configured to execute the computer program stored in the memory to implement a modeling algorithm and an algorithm of a discriminant function as shown in equation (1); the modeling algorithm is a support vector machine algorithm and/or a minimum deviation two-times algorithm;
F(c)＝sgn[f1(c1)+f2(c2)+f3(c3)+f4(c4)+f5(c5)+f6(c6)+f7(c7)+b]
formula (1)
In the formula (1), F (c) represents the effectiveness of the treatment scheme of the brain glioma, F (c) returns that the value is-1 to represent the effectiveness, returns that the value is 1 to represent the ineffectiveness; c. C1、c2、c3、c4、c5、c6And c7Respectively and sequentially showing Ki67 positive rate, microglia positive rate, cytotoxic T cell positive rate, helper T cell positive rate, natural killer cell positive rate, CD31 positive rate and CD105 positive rate; f. of1(c1)、f2(c2)、f3(c3)、f4(c4)、f5(c5)、f6(c6) And f7(c7) Respectively obtaining kernel functions obtained by training according to the modeling algorithm, and b obtaining critical score values obtained by training according to the modeling algorithm;
the output device is used for outputting the effectiveness of the brain glioma treatment scheme.
2. The system of claim 1, wherein the culture medium in step a comprises DMEM/F12 medium, Neurobasal medium Neurobasal, cell culture additive Glutamax, non-essential amino acids NEAAs, cell culture additive B-27 without vitamin a, penicillin-streptomycin, and insulin.
3. The system of claim 2, wherein the DMEM/F12 medium is used in an amount of 0.4-0.6 parts by volume, Neurobasal medium is used in an amount of 0.4-0.6 parts by volume, cell culture additive Glutamax is used in an amount of 0.005-0.02 parts by volume, non-essential amino acids NEAAs are used in an amount of 0.005-0.02 parts by volume, cell culture additive B-27 containing no vitamin a is used in an amount of 0.01-0.04 parts by volume, penicillin-streptomycin is used in an amount of 0.005-0.02 parts by volume, and insulin is used in an amount of 1.5-4 micrograms per milliliter.
4. The system of claim 1, wherein the brain glioma tissue mass has a diameter of 0.3-1 cm.
5. The system according to any one of claims 1 to 4, wherein the amount of the culture medium used in the step a when mixing the brain glioma tissue mass with the culture medium is 1 to 10mL relative to 100mg of the brain glioma tissue mass.
6. The system of any one of claims 1-4, wherein said culturing in step a results in a glioma organoid comprising:
placing the mixture of the brain glioma tissue block and the culture medium in an incubator for shake culture, replacing the culture medium every 2-3 days in the culture process, and continuously culturing for 3-21 days to obtain the brain glioma organoid; wherein the culture conditions comprise: the temperature is 34-40 ℃, and the concentration of carbon dioxide is 4-8%; the shaking speed of the shaking culture is 70-150 rpm.
7. The system of claim 1, wherein in formula (1), f1(c1)＝0.7×c1，f2(c2)＝0.05×c2，f3(c3)＝0.05×c3，f4(c4)＝0.05×c4，f5(c5)＝0.05×c5，f6(c6)＝0.05×c6，f7(c7)＝0.05×c7，b＝-0.15。
8. The system according to claim 1, wherein the microglial positive rate comprises a microglial marker molecule Iba1 positive rate, the cytotoxic T-cell positive rate comprises a cytotoxic T-cell marker molecule CD8 positive rate, and the helper T-cell positive rate comprises a helper T-cell marker molecule CD4 positive rate; the natural killer cell positive rate comprises the positive rate of the marker molecule CD56 of the natural killer cells.
9. A molecular marker for screening a treatment regimen for brain glioma, comprising a marker molecule Ki67 for the level of tumor cell proliferation, a marker molecule Iba1 for microglia, a marker molecule CD8 for cytotoxic T cells, a marker molecule CD4 for helper T cells, a marker molecule CD56 for natural killer cells, a marker molecule CD31 for angiogenesis, and a marker molecule CD105 for angiogenesis.
10. Use of a reagent for detecting molecular markers for the preparation of a kit for screening a treatment regimen for brain gliomas, characterized in that the molecular markers comprise the marker molecule Ki67 for the level of tumor cell proliferation, the marker molecule Iba1 for microglia, the marker molecule CD8 for cytotoxic T cells, the marker molecule CD4 for helper T cells, the marker molecule CD56 for natural killer cells, the marker molecule CD31 for angiogenesis and the marker molecule CD105 for angiogenesis; the reagent for detecting the molecular marker can detect the positive rate of the molecular marker.
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