阅读说明：本技术 膝骨关节炎关节腔内注射治疗的个体化预测方法 (Individualized prediction method for intracavitary injection treatment of knee osteoarthritis ) 是由 金丽霞 姜畅 徐康力 陈子贤 曹渊武 张键 于 2021-09-23 设计创作，主要内容包括：本发明提供了一种膝骨关节炎关节腔内注射治疗的个体化预测方法,该预测方法包括：使用数字评分量表在每次注射前评估患者的分数；通过流式细胞技术,检测关节腔液体的淋巴细胞、T细胞和白细胞数；第一次和第二次关节腔注射前,若数字评分量表的评分改善了2分或以上,则继续治疗；若数字评分量表的评分改善了1分,且淋巴细胞/白细胞比例增加,则继续治疗；若数字评分量表的评分无改善,淋巴细胞/白细胞比例增加且T细胞/白细胞比例增加,则继续治疗；本发明能够个体化预测治疗效果,在进行完整治疗周期之前,提前辨别出治疗效果不好的患者,及早建议其改行其他治疗方案,以减少关节腔内注射相关并发症,并促进医疗资源合理使用。(The invention provides an individualized prediction method for knee osteoarthritis joint intracavity injection treatment, which comprises the following steps: evaluating the patient's score prior to each injection using a numerical scoring scale; detecting the number of lymphocytes, T cells and white cells of the joint cavity liquid by a flow cytometry technology; continuing treatment if the score on the numerical score scale is improved by 2 or more points prior to the first and second joint cavity injections; if the score on the numerical score scale improves by 1 point and the lymphocyte/leukocyte ratio increases, treatment continues; if there is no improvement in the score on the numerical score scale, the lymphocyte/leukocyte ratio is increased and the T cell/leukocyte ratio is increased, then treatment is continued; the invention can individually predict the treatment effect, distinguish patients with poor treatment effect in advance before carrying out a complete treatment cycle, and recommend other treatment schemes to be changed as soon as possible so as to reduce complications related to injection in joint cavities and promote reasonable use of medical resources.)

1. An individual prediction method for knee osteoarthritis joint intracavity injection treatment is characterized by comprising the following steps: which comprises the following steps:

1) and pain assessment: using a numerical scoring scale, assessing the score of the patient prior to each injection;

2) and detecting joint cavity liquid: detecting the number of lymphocytes, T cells and white cells of the joint cavity liquid by a flow cytometry technology;

3) scoring according to a numerical scoring scale, and analyzing the cellular components of the articular cavity fluid.

2. The method for individualized prediction of intracavitary injection therapy for knee osteoarthritis according to claim 1, characterized in that: in step 3), the individualized prediction is as follows: prior to the first and second joint cavity injections, the treatment is continued if the score on the numerical scale improves by 2 points or more.

3. The method for individualized prediction of intracavitary injection therapy for knee osteoarthritis according to claim 1, characterized in that: in step 3), the individualized prediction is as follows: if the score on the first and second numerical scoring scale improves by 1 point and the lymphocyte/leukocyte ratio increases prior to the first and second joint cavity injections, treatment is continued, otherwise treatment is discontinued.

4. The method for individualized prediction of intracavitary injection therapy for knee osteoarthritis according to claim 1, characterized in that: in step 3), the individualized prediction is as follows: if the score on the first and second numerical scale is not improved, the lymphocyte/leukocyte ratio is increased and the T cell/leukocyte ratio is increased prior to the first and second joint cavity injections, the treatment is continued, otherwise the treatment is abandoned.

5. The method for individualized prediction of intracavitary injection therapy for knee osteoarthritis according to claim 1, characterized in that: in step 1), 0-10 in the numerical rating scale represent pain of different degrees, 0 is no pain, and 10 is severe pain.

Technical Field

The invention belongs to the technical field of medicine, and particularly relates to an individualized prediction method for the intra-articular injection treatment effect of knee osteoarthritis.

Background

Knee Osteoarthritis (KOA) is a common osteoarticular disease which is frequently seen in old people, and the knee swelling and pain is a remarkable clinical manifestation, which seriously affects the health and life quality of the old people. Symptomatic knee osteoarthritis is present in 10% of men and 13% of women over the age of 60.

Current treatments for KOA are based on conservative treatment to delay the time of knee replacement by alleviating local symptoms. Intra-articular injection is the main conservative treatment of KOA. The currently used injections include sodium hyaluronate, glucocorticoid, chitosan, Platelet Rich Plasma (PRP), autologous isolated stem cells, and the like. The injection in the joint cavity needs to be repeated, the general treatment course is once a week for 5 weeks, and the effective rate is 60-70%. The multiple injections in the joint cavity bring pressure for patients to see and treat, and also increase the risks of infection and bleeding in the joint cavity.

Disclosure of Invention

Aiming at the defects in the prior art, the invention aims to provide an individualized prediction method for knee osteoarthritis joint intracavity injection treatment.

In order to achieve the above purpose, the solution of the invention is as follows:

an individualized prediction method for knee osteoarthritis joint intracavity injection treatment, which comprises the following steps:

1) and pain assessment: using a numerical scoring scale (NRS), the patient's score is assessed prior to each injection;

2) and detecting joint cavity liquid: detecting the number of lymphocytes, T cells and white cells of the joint cavity liquid by a flow cytometry technology;

3) scoring according to a numerical scoring scale, and analyzing the cellular components of the articular cavity fluid.

As an embodiment of the present invention, in step 3), the individualized prediction is: NRS was used for scoring before the first and second joint cavity injections, and treatment continued if NRS score improved by 2 points or more.

As an embodiment of the present invention, in step 3), the individualized prediction is: if the first and second NRS scores improved by 1 point and the lymphocyte/leukocyte ratio increased prior to the first and second joint cavity injections, treatment is continued, otherwise treatment is discontinued.

As an embodiment of the present invention, in step 3), the individualized prediction is: if the first and second NRS scores do not improve, the lymphocyte/leukocyte ratios increase and the T cell/leukocyte ratios increase before the first and second joint cavity injections, the treatment is continued, otherwise the treatment is abandoned, e.g. if the lymphocyte/leukocyte ratios decrease or the T cell/leukocyte ratios decrease.

As an embodiment of the present invention, in step 1), 0-10 in NRS represents pain of various degrees, 0 is no pain, and 10 is severe pain.

Due to the adoption of the scheme, the invention has the beneficial effects that:

the invention can individually predict the intracavity injection treatment effect of the knee osteoarthritis patient, distinguish the patient with poor treatment effect in advance before carrying out the complete treatment period, propose other treatment schemes to change as soon as possible, and reduce the injection times from the original 5 times to 2 times, thereby helping the clinician to make treatment decisions, reducing the related complications of the intra-articular injection, promoting the reasonable use of medical resources and improving the use efficiency of the medical resources.

Drawings

FIG. 1 is a flow chart of the individual prediction method for injecting the treatment into the joint cavity of knee osteoarthritis according to the present invention.

FIG. 2 is a schematic view of flow cytometry detection of immunocytes in example 1 of the present invention.

Detailed Description

The invention provides an individualized prediction method for intracavitary injection treatment of knee osteoarthritis joints, so that a clinician is helped to decide whether to complete the full-course operation of intracavitary injection.

The individualized prediction method for the knee osteoarthritis joint intracavity injection treatment comprises the following steps:

1) evaluation of pain: using the NRS scale, patients were assessed for NRS score prior to each injection.

NRS pain score refers to a numerical rating scale by using 0-10 to represent varying degrees of pain, 0 being no pain, 10 being the most severe pain, asking the patient how severe the pain is, or allowing the patient to self-circle a number that best represents the level of pain itself.

2) Detection of joint cavity liquid: the lymphocyte, T cell and leukocyte counts of the joint cavity liquid are detected by the flow cytometry.

3) The personalized prediction method comprises the following steps: as shown in fig. 1.

Scoring using the NRS assessment scale prior to the first and second joint cavity injections, and continuing treatment if the NRS score improves by 2 points or more.

Ii) if the first and second assessment of NRS score improved by 1 point, then the first and second effusion compartment are analyzed for changes in lymphocyte to leukocyte ratios, and if the lymphocyte to leukocyte ratio increased, treatment is continued, and if the ratio decreased, the treatment regimen is not recommended to continue because of its lesser potential benefit.

And iii) if there is no improvement in NRS scores from the first and second assessments, analyzing the first and second articular cavity droppings for changes in lymphocyte and T cell leukocyte ratios, and continuing treatment if the lymphocyte/leukocyte ratio is increased and the T cell/leukocyte ratio is increased, otherwise not recommending continuation of the treatment regimen because of its potential benefit is less.

The present invention will be further described with reference to the following examples.

Example 1:

1) instruments and reagents

The flow cytometer used was FACSCANTIII and FACSLYRic purchased from BD, the centrifuge, the micro-sample application gun and the 1.5mL Ep tube were purchased from Eppendorf, Germany, the refrigerator was purchased from Haier, Qingdao, the disposable pipette and the flow tube (specification: 12X 75) were purchased from Kyoto, Zhejiang, and the horizontal shaker, the suction filter and the 1 XPBS solution were prepared by itself. Or the same kind of instruments and reagents with the same functions.

2) Sample collection and processing

According to the routine articular cavity injection treatment process, the external upper edge of the patella is selected to insert a needle, the needle needs to be withdrawn before injection to determine whether the patella is in the articular cavity, and partial articular cavity effusion is extracted to relieve the articular cavity pressure. The articular cavity fluid obtained in this process was used as a specimen for sampling. Extracting the joint cavity effusion by a needle cylinder for 2mL, adding the joint cavity effusion into an EDTA anticoagulation tube, storing the joint cavity effusion in a refrigerator at 4 ℃, and treating the joint cavity effusion within 24 h. When in treatment, the EDTA anticoagulation tube filled with joint cavity hydrops is placed at 1500rpm for centrifugation for 5min, and supernatant is discarded; transferring the residual liquid to a flow tube, and carrying out immune cell detection after washing.

3) Immune cell detection

Adding 2mL of 1 XPBS solution into a flow tube with 1mL of joint cavity effusion, uniformly blowing and beating the solution by using a disposable pipette, centrifuging the solution at 1500rpm for 5min, and washing the solution twice. After discarding the supernatant, the cells were resuspended. To the resuspended cell fluid was added 2. mu.L of the following antibody combination (Table 1):

TABLE 1

Fluorescent channel	BV421	V500
			Name of label	CD3	CD45
Dosage of mu L	1	1

After mixing the cells and the antibody, the mixture was mixed well and incubated for 15min at room temperature in the dark. During the incubation period, the Lysing Solution 10 × Concentrate was diluted to 1 × erythrocyte lysate, 1mL of 1 × erythrocyte lysate was added to the cell suspension after the incubation was completed, mixed well, and incubated at room temperature in the dark for 10 min. After centrifugation at 1500rpm for 5min, the cells were washed once with 1 XPBS solution. After discarding the supernatant, it was resuspended in 100. mu.L of 1 XPBS solution and ready for detection on a flow machine.

4) Flow cytometer detection and analysis

Immune cells were harvested using a BD FACSCAnto II flow cytometer and BD FACS Diva 8.0 software. As shown in FIG. 2, cell debris and adherent cells and debris were removed by SSC-A/FSC-A, FSC-H/FSC-A, and SSC-A/CD45 was gated to recognize Leukocytes (Leukocytes) and Lymphocytes (Lymphocytes), and SSC-A/CD3 was gated to recognize T cells (T cells). All cells were collected and the total number of leukocytes, lymphocytes and T cells were determined. BD FACS Diva 8.0 software was used for data analysis of immune cells.

Example 2:

based on a prospective observational cohort study, a total of 18 patients were enrolled, with inclusion criteria: patients undergoing intra-articular injection therapy for knee osteoarthritis; the age is not limited, and the nature is not limited; when the joint cavity is injected, a certain amount of joint fluid can be extracted; it is expected that 5 joint cavity injections (a conventional course of treatment including 5 aspirations and joint cavity injections) can be completed in our hospital; subjects signed the book for informed consent. The effective number of people is 13, and the effective rate is 72.2%. NRS pain scores from 5 visits of the 18 patients and cytokine and flow cytometry analysis of joint cavity effusion for 86 total times were collected and analyzed, and statistical methods such as logistic analysis gave the following effective rate data.

1) Prior to treatment by intra-articular cavity injection, patients were assessed for pain using the NRS assessment scale.

2) According to the conventional intra-articular injection treatment process, before injection, the joint cavity needs to be withdrawn to determine whether the joint cavity is in the articular cavity, and partial effusion of the articular cavity is withdrawn to relieve the pressure of the articular cavity.

3) Performing flow cytometry on the effusion of the knee joint cavity: the total number of leukocytes, number of lymphocytes and number of T cells were determined.

4) The following individualized predictions were made:

a) if the NRS score is improved by 2 points or more before the first and second articular cavity injections, and early data suggest that the improvement effective rate reaches 99%, the treatment is continued.

b) If the NRS score is improved by 1 point before the first and second joint cavity injections, the change of the proportion of lymphocytes in the white blood cells in the joint cavity effusion for the first and second times needs to be analyzed, and if the proportion of the lymphocytes/white blood cells is increased, early data suggest that the improvement efficiency is over 99 percent. If the ratio is reduced, earlier data suggests that the improvement efficiency is less than 1%.

c) If the NRS is not improved in the first and second evaluations, the first and second articular cavity dropsy are analyzed for changes in the proportion of lymphocytes and T cells in the leukocytes, and if the lymphocyte/leukocyte ratio is increased and the T cell/leukocyte ratio is increased, the prophase data suggest that the improvement efficiency rate is over 60%, otherwise the efficiency rate is less than 1%.

The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. It will be readily apparent to those skilled in the art that various modifications to these embodiments and the generic principles defined herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above-described embodiments. Those skilled in the art should appreciate that many modifications and variations are possible in light of the above teaching without departing from the scope of the invention.