Blood marker for assisting diagnosis of limb nerve captive pressure injury

Abstract

The invention belongs to the field of biological medicine industry, and discloses a blood marker for assisting diagnosis of limb nerve captive pressure injury. The inventor verifies that the LRG1 is the peripheral blood marker of the limb nerve entrapment injury for the first time, the average value of the LRG1 in blood is higher than that of a normal person when the limb tumor causes the nerve entrapment, the LRG1 is more obviously increased along with the aggravation of the injury, and the LRG1 serving as the peripheral blood marker of the limb tumor causes the nerve entrapment has higher sensitivity and specificity. The invention proposes and verifies that the LRG1 is the peripheral blood marker of the nerve entrapment caused by the limb tumor for the first time, and the result can be used for auxiliary diagnosis of the nerve entrapment caused by the limb tumor.

Description

Technical Field

[0001] The present invention belongs to the field of the biomedical industry, and particularly relates to the application of an LRG1 quantitative reagent in the preparation of a reagent for detecting limb nerve entrapment injury. Background Art

[0002] 1. Compression injuries of limb nerves are common in carpal tunnel syndrome, cubital tunnel syndrome, supinator syndrome, pronator teres syndrome, piriformis syndrome, common peroneal nerve entrapment syndrome, and tarsal tunnel syndrome, etc., among which compression of peripheral nerves in the upper limb is more common.

[0003] 1.1 Carpal Tunnel Syndrome The carpal tunnel is located at the root of the palm. The bottom and both sides are formed by the carpal bones, and the transverse carpal ligament spans across it, forming a fibro-osseous canal. Carpal tunnel syndrome (CTS), commonly known as mouse hand, is a peripheral nerve entrapment syndrome caused by compression of the median nerve in the carpal tunnel. The main manifestations are pain in the front of the wrist and numbness and weakness in the hand, commonly seen in the radial 3.5 fingers of the median nerve distribution.

[0004] 1.2 Pronator Teres Syndrome The median nerve is compressed by the tendinous arch between the two heads of the pronator teres at the proximal end of the forearm. It is more common when the forearm rotates strongly repeatedly. The main manifestations are pain in the proximal end of the forearm, mainly pain in the pronator teres area, which can radiate to the elbow, upper arm, neck, and wrist. Numbness in the radial side of the palm and the radial 3.5 fingers, weakened pinch force of the thumb and index finger, flexion of the metacarpophalangeal joint of the thumb and the proximal interphalangeal joint of the index finger, and hyperextension of the distal interphalangeal joint when the thumb and index finger oppose each other, with mild atrophy of the thenar muscle.

[0005] 1.3 Supinator Syndrome Supinator syndrome is caused by compression of the deep branch of the radial nerve (posterior interosseous nerve) near the tendinous arch of the supinator muscle. The clinical manifestations are dysfunction of the extensor muscles of the forearm; difficulty in abducting and extending the thumb, and inability of the metacarpophalangeal joints of the 2nd - 5th fingers to extend actively; the wrist joint can extend actively (the radial wrist extensors are not innervated by the deep branch of the radial nerve), and there is no sensory abnormality in the tiger's mouth area.

[0006] 1.4 Cubital Tunnel Syndrome Two bony prominences can be felt on the posteromedial side of the elbow joint (i.e., between the medial epicondyle of the humerus and the olecranon of the ulna). There is an arc-shaped narrow and deep bony groove between the two, and there is a fibrous sheath tube - the ulnar nerve groove inside it, in which a tough cord-like structure can be felt, that is, the ulnar nerve. Around the elbow joint, the ulnar nerve runs in a tunnel called the cubital tunnel (ulnar cubital tunnel). Narrowly defined, the cubital tunnel refers to the ulnar nerve groove, while in a broad sense, the scope of the cubital tunnel can extend from the Struthers arch on the medial side of the upper arm (about 8 cm from the medial epicondyle) to the proximal part of the forearm (between the ulnar and humeral heads of the flexor carpi ulnaris). Compression, traction, or friction of the ulnar nerve in this channel will produce a series of symptoms and signs. Common clinical manifestations include paresthesia or numbness in the ring and little fingers. In severe cases, there will be weakness of hand strength, muscle atrophy, decreased flexibility, and inability to perform fine movements, etc.

[0007] 1.5 Ulnar tunnel syndrome at the wrist Ulnar tunnel syndrome at the wrist, also known as Guyon canal syndrome, pisohamate hiatus syndrome, Ramsay-Hunt syndrome. The cross-section of the ulnar tunnel at the wrist is triangular. The anterior wall is the superficial transverse carpal ligament, the posterior wall is the deep transverse carpal ligament, and the medial wall is the pisiform bone and pisohamate ligament. The ulnar nerve and the ulnar artery and vein pass through it. Compression of the ulnar nerve inside it causes ulnar tunnel syndrome. Involvement of the superficial branch causes sensory disturbance in the area innervated by the ulnar nerve. Compression of the deep branch can lead to atrophy and weakness of the intrinsic muscles of the hand, deep pain and burning pain in the hand, significant nocturnal pain, adduction of the thumb, weakness in abduction and adduction of the other four fingers, and the ring and little fingers may show claw-shaped deformities. The paper pinch test and Froment test are positive.

[0008] 1.6 Common peroneal nerve entrapment syndrome This disease is caused by entrapment of the common peroneal nerve in the osteofascial canal of the fibular neck. Injury and external compression are common causes. It is manifested as pain, numbness, and motor disorders in the lateral side of the foot and calf. Weakness in dorsiflexion of the ankle and extension of the toes, and weakness or disappearance of eversion force. There may be sensory disturbance in the lateral side of the calf and the lateral side of the foot, and there may be tenderness and Tinel sign at the fibular neck.

[0009] 1.7 Tarsal tunnel syndrome Also known as the plantar tunnel syndrome or the tarsal tunnel syndrome, it refers to a series of clinical symptoms and signs caused by entrapment of the tibial nerve during its course through the tarsal tunnel located posterior and inferior to the medial malleolus to the sole of the foot. The onset of the disease in patients is slow and mostly occurs on one side. In the early stage, it is manifested as intermittent pain, tightness, swelling discomfort, or numbness in the sole of the foot and the heel. The pain sometimes radiates to the calf and sometimes there are twitches along the foot arch. It is aggravated after standing or walking for a long time, and there is a history of waking up due to pain at night. Most patients can get relief after taking off their shoes.

[0010] 2. Mechanism of peripheral nerve entrapment injury, mechanism of nerve entrapment injury: One end of the peripheral nervous system is connected to the brain or spinal cord of the central nervous system, and the other end is connected to the organs of various systems of the body. A nerve is composed of nerve fibers, which are composed of the long processes of neurons and a part of the glial cells that surround them. The processes of Schwann cells, which belong to glial cells, surround the axons of neurons to form a myelin sheath with a concentric lamellar structure. The myelin sheath has a certain insulating and supporting effect, ensuring the high-speed conduction of electrical signals along the axons. According to whether nerve fibers have a myelin sheath, they can be divided into myelinated fibers and unmyelinated fibers.

[0011] In addition to the neurilemma and myelin sheath formed by Schwann cells, nerve fibers are also surrounded by connective tissue to form the endoneurium, which contains a fibrous matrix and fibroblasts. Many nerve fibers are bundled together, and the outside is surrounded by a relatively fine layer of connective tissue called the perineurium. The perineurium generally contains 15 - 20 layers of cells and is a metabolically active diffusion barrier.

[0012] Nerve bundles of different thicknesses are concentrated to form a nerve, and the outside is surrounded by an epineurium composed of a loose connective tissue. The epineurium contains collagen fibers, fibroblasts, fat, lymphatic vessels, blood vessels, and nerves (i.e., the nerves of the nerve).

[0013] Blood vessels of the nerve: The blood vessels of the nerve are divided into an extrinsic system and an intrinsic system.

[0014] Extrinsic system: That is, the local nutritive blood vessels and epineurial blood vessels. They originate from the accompanying blood vessels of adjacent tissues. At regular intervals, they give off segmental blood vessels to the epineurium and then divide into ascending and descending branches. The ascending and descending branches of adjacent segmental blood vessels anastomose with each other to form longitudinally arranged epineurial blood vessels.

[0015] The epineurial blood vessels give off short transverse or oblique branches that pass through the epineurium to the interbundle region to form interbundle blood vessels. The branches of the interbundle blood vessels pass through the perineurium and enter the bundle to form intrafascicular microvessels and finally reach the endoneurium to complete the blood supply of the nerve fibers.

[0016] Intrinsic system: It refers to the longitudinally running microvascular network within the endoneurium. The two vascular systems have rich anastomoses. The vascular smooth muscle of the intrinsic system is poorly developed and lacks self-regulatory function. There are nerve plexuses on the walls of the blood vessels of the extrinsic system, which have a certain regulatory function on the blood vessels.

[0017] There is nerve tissue in the connective tissue (epineurium) of the peripheral nerve. These nerves of the nerve are also called nervi nervorum. The function of the nervi nervorum is to ensure the intrinsic sensitivity of the nerve, that is, it can regulate external mechanical stimuli (such as nerve compression) and its own metabolic nutrition. In addition, the outer membrane of the blood vessels of the nerve is also innervated, which can be used to regulate the contraction and relaxation of the blood vessels.

[0018] Mechanical properties of the nerve Motility: It refers to the fact that the activities of the body exert various forces on nerve tissues. Nerves have the property of adapting to external forces. For example, within the path (duct) through which they pass, they adapt in a sliding manner; they change their shape (wrinkling, twisting, folding, etc.) to reduce external force damage and withstand a certain degree of stretching.

[0019] Viscoelasticity: When an object with viscoelastic properties is subjected to an external force, it can immediately adapt by deforming; however, after the external force is removed, it still retains a part of the deformation. Elastic objects are different. These objects will immediately return to their original shape after the external force is removed.

[0020] Nerve tissue is a structure with viscoelastic properties. When a nerve is subjected to tensile external forces, the viscoelastic properties can provide more adaptability to the nerve tissue, so that it will not break even if it is overstretched. When encountering compressive external forces, it is very easy to cause damage to the nerve, and even a small force will have the same effect.

[0021] The internal tension environment of nerve tissue Endogenous intraneural pressure represents the sum of all intracellular (axonal hydrostatic pressure); this hydrostatic pressure is further increased by the blood pressure in the nervi vasorum; the viscoelastic properties make nerve tissue prone to exposure to longitudinal, centrifugal, and distal-directed tensions; as long as the intensity of the tension changes, the endogenous intraneural pressure will immediately change accordingly. Exogenous intraneural pressure acts on various sheaths that surround the nerve.

[0022] The relative relationship between tension and pressure: Directly compressing nerve tissue will cause a sharp increase in tissue tension; applying a longitudinal pull to a already tense nerve tissue will reduce it.

[0023] Peripheral nerve injuries include endogenous nerve injuries: fibrosis, hematoma, or endogenous pressure of nerve tissue, primary local ischemia, and obstruction of lymphatic and venous return. Exogenous nerve injuries: Nerves are prone to entrapment injuries when passing through osseofibrous canals, or entering and exiting the deep fascia, or passing through the interior of muscles, etc. For example, the median nerve in the carpal tunnel, the median nerve under the pronator teres, the musculocutaneous nerve passing through the coracobrachialis, and the superficial peroneal nerve emerging from the deep fascia at the middle and distal 1 / 3 of the calf are all areas where nerves are prone to entrapment. Exogenous nerve entrapment: According to the nature of the entrapment, it can be divided into acute nerve entrapment and chronic nerve entrapment.

[0024] Acute nerve compression can cause obstruction of the blood vessels within the nerve, affecting nerve function, and also mechanically damage the nerve structure. After chronic nerve compression, the epineurium thickens, myelinated nerves undergo demyelination changes, and even nerve fiber degeneration may occur (also known as secondary degeneration, which refers to a series of degenerative changes and phagocytosis processes such as axonal necrosis, myelin sheath decomposition and disappearance, and proliferation of the neurilemma at the distal end after axonal injury due to blocked axoplasmic transport). In addition, after nerve compression, the motor nerve conduction velocity first increases, and as the disease progresses, the nerve conduction velocity slows down.

[0025] Multiple nerve compression syndrome refers to the situation where a nerve is compressed at multiple sites during its course. Each individual compression is not sufficient to produce any symptoms, but together they can manifest as nerve compression symptoms.

[0026] In 1973, Upton and Mccomas clinically observed evidence of cervical nerve root lesions in carpal tunnel syndrome and ulnar nerve lesions. They also pointed out that diabetes is prone to carpal tunnel syndrome due to multiple nerve compressions: each individual compression is not sufficient to produce any symptoms, but together they may cause symptoms. For example, the transverse carpal ligament already compresses the median nerve, but there are no symptoms. As age increases, once cervical spondylosis or thoracic outlet syndrome appears, typical carpal tunnel syndrome manifestations will occur. Therefore, it is very necessary for us to understand the compression sites and their fixed points of the nerve during its course.

[0027] 3. Impact after nerve injury When a peripheral nerve presents with single or multiple compression syndromes during its course, mild nerve injury symptoms often occur. If the nerve injury is mild and the nerve is irritated, the muscles it innervates show increased tension, or increased tension in some muscle fibers (cords can be palpated within the muscle). Its muscle strength may increase in the early stage and decrease in the later stage.

[0028] The skin sensation in the nerve innervation area is sensitive, decreased, or painful. In addition, after nerve injury, the impairment of proprioceptive afferentation in the body can lead to joint or body instability. The motor perception system maintains the balance of joints and the body. If sensory nerve fibers are damaged, resulting in reduced, increased, or abnormal information transmitted to the central nervous system, abnormal efferent manifestations such as joint or body instability can occur. If motor nerve fibers are damaged, even if the body signals are transmitted normally, abnormal efferent manifestations will also occur, manifested as joint or body instability.

[0029] 4. Clinical manifestations According to the different sites of nerve compression and nerve structures, the symptoms manifested by compression at different sites are also different. The common symptoms are mainly: Pain is often manifested as numb pain in the innervated area. It usually hurts most at night or in the early morning, and there may be radiating pain. The pain can be relieved after appropriate activities. For example, in carpal tunnel syndrome, there is often numb pain and weakness at the fingertips of the thumb, index finger and middle finger first, and it can be relieved after shaking the wrist appropriately. Sensory disturbance, manifested as sensory loss or abnormality in the innervated area.

[0030] Deformity: Hand deformities may occur in some nerve compressions. For example, in cubital tunnel syndrome, atrophy of the hypothenar and interosseous muscles occurs, and the ring and little fingers show claw-like deformities. In supinator syndrome, the wrist joint can be actively extended, but it deviates to the affected side, and the extensor function of the forearm is impaired.

[0031] Motor disturbance: The nerve motor function is damaged, manifested as difficulty in holding objects and limping when walking. For example, in supinator syndrome, abduction and extension of the thumb are impaired, and the metacarpophalangeal joint cannot be actively extended. In piriformis syndrome, there may be painful limping and mild atrophy of the calf muscles.

[0032] 5. Diagnosis of peripheral nerve compression: It usually involves medical history taking, physical examination, and necessary imaging examinations and electrophysiological tests to confirm the location and severity of nerve compression.

[0033] Medical history taking: Doctors will ask patients in detail about their symptoms, duration, inducing factors, etc. Common symptoms include pain, numbness, tingling or weakness, and these symptoms may worsen or relieve with activity.

[0034] Physical examination: By palpation, observing movements and performing specific position tests, doctors can evaluate nerve function. For example, carpal tunnel syndrome can be detected by Tinel's sign or Phalen's test.

[0035] Imaging examination: Imaging methods such as X-ray, ultrasound, CT or MRI can help identify structural problems causing nerve compression, such as skeletal deformities, tumors or cysts.

[0036] Electrophysiological examination: Electromyography and nerve conduction velocity tests can measure the electrical activities of nerves and muscles, and evaluate the degree and location of nerve damage. These tests are very important for determining the specific nature of nerve damage.

[0037] Through the above steps, the cause and location of nerve compression can be determined more accurately, providing guidance for subsequent treatment. Early identification and intervention help improve symptoms and prevent long-term damage.

[0038] 6. Prevention and treatment Nerve compression caused by tumors can generally be solved by physical therapy, drug therapy, surgical treatment, etc.

[0039] Physical therapy: It may belong to a benign tumor, and the tumor is relatively large, pressing on the nerve. Physical therapy such as massage, acupuncture, etc. can be used to improve the condition.

[0040] Drug therapy: If the nerve compressed by the tumor shows pain, under the guidance of a doctor, pain-relieving and nerve-nourishing drugs such as ibuprofen tablets and oryzanol tablets can be taken for treatment.

[0041] Surgical treatment: If the tumor is large and continuously presses on the nerve, it may have a certain impact on physical health. Therefore, it is necessary to promptly treat it by surgically removing the tumor.

[0042] 7. Relationship with proteins in the blood: Proteins are the executors of gene functions. The study of protein structure, localization, and protein-protein interactions will provide a direct basis for clarifying the essence of life phenomena. Almost all physiological and pathological processes, as well as the actions of drugs and environmental factors, depend on proteins and cause changes in the proteome. Before any disease shows any detectable symptoms, certain proteins must have changed. Therefore, finding the key proteins and biomarker proteins for various diseases is of great significance for disease diagnosis, pathological research, and drug screening. Leucine-rich repeat family proteins, which include eight leucine-rich repeats (LRR), have been shown by research to be transmembrane proteins. They mainly participate in normal physiological activities as cell adhesion molecules or ligand-binding proteins.

[0043] Leucine-rich α2-glycoprotein 1 (LRG1), is a member of the leucine-rich repeat family. LRG1 belongs to the LRR protein family. Past studies have shown that LRG1 is involved in important physiological and pathological processes of the body, such as protein interaction, signal transduction, and cell adhesion.

[0044] Currently, a variety of proteins with leucine-rich repeat structures have been found in the nervous system. In some neurodegenerative diseases, a decrease in the concentration of LRG1 in cerebrospinal fluid has been detected, and the expression of human LRG1 has been observed in the dissected cerebral cortex. In past studies, some scholars performed proteomic analysis on cerebrospinal fluid specimens of normal individuals and patients with idiopathic normal pressure hydrocephalus (INPH), and found that the expression of LRG1 was specifically increased in the cerebrospinal fluid of INPH patients. Thus, LRG1 has potential value for the clinical diagnosis of INPH. The abnormal expression of LRG1 is associated with the occurrence of various tumors, such as glioma, lung cancer, colon cancer, epithelial ovarian cancer, hepatocellular carcinoma, etc.

[0045] In the process of studying retinopathy caused by vascular remodeling, it was found that LRG1 can promote the mitosis of endothelial cells and the neovascularization of blood vessels. This hypothesis was further confirmed in the in vitro lumen formation assay of human vascular endothelial cells (HUVEC). By providing recombinant human LRG1 as a medium, a significant increase in lumen formation and branching can be caused, while anti-LRG1 antibody significantly blocks the formation of lumens. Another study shows that the angiogenic activity of LRG1 is not limited to the eyes and may also play an important role in other major biological processes, including tumors and immune responses, etc. The regulatory effect of LRG1 on the TGF-β1 signaling pathway provides strong evidence for studying the function of LRG1. LRG1 is a very promising therapeutic target for controlling ocular angiogenic diseases and may also play an important role in other diseases such as cancer and atherosclerosis.

[0046] The function of LRG1 may be related to its LRR structure. The function of LRG is considered to play a regulatory role by binding to other proteins such as cytochrome C, thereby restricting the functions of some proteins. Gliomas originate from glial cells and are the most common tumors in the central nervous system, accounting for 30% of all brain and central nervous system tumors and 80% of all malignant brain tumors. Related experiments have shown that LRG1 plays an important role in promoting the proliferation, migration, and invasion of glioma cells, and these functions are achieved through mediating the transforming growth factor β1 (TGF-β1) signal transduction pathway. Moreover, LRG1 can promote endothelial cell proliferation and angiogenesis through the TGF-β signaling pathway, suggesting that LRG1 may be involved in the occurrence and development of astrocytic malignancies. Previous studies have shown that inhibiting the expression of LRG1 inhibits the in vitro growth of glioma cells, and in xenograft mouse models, it delays the occurrence of gliomas, providing evidence for the tropic effect of LRG1 on glioma pathological changes. LRG1 is related to atherosclerosis and microcirculation disorders and may be used as a marker of vascular injury in different vascular beds. LRG1 has a potential role in the early identification of the risk of vascular diseases, and the pathological basis and mechanisms of these correlations still need to be further explored. It can be considered that altering the activity of LRG1 has therapeutic potential in vascular diseases, especially in minimally invasive surgery and endovascular revascularization. The expression of LRG1 is upregulated after myocardial infarction. As a novel cardiovascular protective factor that inhibits cardiac remodeling after myocardial infarction, LRG1 may become a therapeutic target for cardiovascular diseases. Gene deletion of LRG1 exacerbates cardiac dysfunction after myocardial infarction and increases myocardial fibrosis. In LRG1-deficient mice, the smad1 / 5 / 8 pathway is impaired, resulting in a decrease in capillary density at the infarct border. Gene ablation of LRG1 exacerbates the reduction in capillary density after myocardial infarction in the cardiac remodeling model. In addition, our study shows that the level of LRG1 in the blood increases after cardiopulmonary cerebral resuscitation in patients with cardiac arrest, which may be related to cerebral ischemia, brain injury, and vascular injury caused by cardiac arrest, consistent with the mechanism of increased LRG1 in the blood of patients with cerebral infarction and myocardial infarction.

[0047] On the other hand, as an angiogenic factor, LRG1 may have promising prospects for the treatment of vascular diseases by regulating the smad1 / 5 / 8 pathway, but its specific regulatory mechanism is still unclear.

[0048] The prior Chinese patent application CN117110626A of the inventor discloses that when there is a neck artery lesion, the average value of LRG1 in the blood is higher than that of normal people, and as the lesion worsens, the shedding of plaques in the neck blood vessels increases more significantly. When used as a peripheral blood marker for dangerous neck artery lesions, it has high sensitivity and specificity and can be used in combination with the results of B-ultrasound and other methods for diagnosing neck artery lesions. CN115856325A discloses that the content of LRG1 in peripheral blood is closely related to lower limb artery injury. By detecting the content of LRG1 in peripheral blood, it is possible to well predict lower limb artery injury, especially lower limb arteriosclerosis or plaques, lower limb artery stenosis, thrombosis or occlusion, and especially can well predict lower limb arteriosclerosis or plaques, lower limb artery stenosis, thrombosis or occlusion caused by hypertension, hyperlipidemia or hyperglycemia. CN110261617A discloses that LRG1 can be used as a peripheral blood marker for intracerebral hemorrhage.

[0049] In the prior art, there is no research indicating that LRG1 is related to limb nerve compression injury. Summary of the Invention

[0050] The purpose of the present invention is to overcome at least one deficiency of the prior art and provide the use of an LRG1 quantitative reagent in the preparation of a reagent for detecting limb nerve compression injury.

[0051] The technical solution adopted by the present invention is: In the first aspect of the present invention, there is provided: The use of an LRG1 quantitative reagent in the preparation of a reagent for diagnosing or assisting in the diagnosis of limb nerve compression injury.

[0052] In some examples, the LRG1 quantitative reagent is an ELISA reagent, an immunofluorescence reagent, or a mass spectrometry analysis reagent.

[0053] In some examples, the sample detected by the LRG1 quantitative reagent is peripheral blood.

[0054] In some examples, the limb nerve compression injury is a limb nerve compression injury caused by a tumor.

[0055] In the second aspect of the present invention, there is provided: A system for diagnosing or assisting in the diagnosis of limb nerve compression injury, comprising: An LRG1 quantification device for quantifying the amount of LRG1 in a sample; An analysis device for determining the risk of limb nerve compression injury based on the content of LRG1; A result output device for outputting the analysis result of the analysis device.

[0056] In some examples, the LRG1 quantification device is an ELISA detection device, an immunoblotting device, an immunofluorescence device, or a mass spectrometry device.

[0057] In some examples, a high level of LRG1 indicates a high risk of limb nerve entrapment injury.

[0058] In some examples, a high level of LRG1 means that its content in peripheral blood samples is higher than 5 ng / mL.

[0059] In some examples, the sample is peripheral blood.

[0060] In some examples, the limb nerve entrapment injury is a limb nerve entrapment injury caused by a tumor.

[0061] The beneficial effects of the present invention are as follows: The inventors first studied and confirmed that LRG1 is a peripheral blood marker for limb nerve entrapment injury. The average value of LRG1 in the blood is higher in nerve entrapment injury than in normal people, and it increases more significantly with the aggravation of the injury. As a peripheral blood marker for nerve entrapment caused by limb tumors, it has high sensitivity and specificity.

[0062] The test results show that with a peripheral blood LRG1 content of 5 ng / ml as the CUTOFF value, among 30 patients with limb nerve entrapment injury, 28 cases were successfully detected, 2 cases were false negatives, and the sensitivity was 93.33%; among 60 normal people, 12 were false positives, and the specificity was 80%. After surgery for limb nerve entrapment injury, LRG1 decreased significantly compared with before surgery, P<0.001, and was comparable to the level of normal people, P>0.05. Detailed implementation manners

[0063] The first aspect of the present invention provides: The application of an LRG1 quantitative reagent in the preparation of a reagent for diagnosing or assisting in the diagnosis of limb nerve entrapment injury.

[0064] The LRG1 quantitative reagent can be various conventional protein quantitative reagents without special requirements. In some examples, the LRG1 quantitative reagent is an ELISA reagent, an immunofluorescence reagent, or a mass spectrometry analysis reagent. The LRG1 quantitative reagent is preferably an ELISA reagent, which is convenient to operate and the results can meet the requirements.

[0065] In some examples, the sample detected by the LRG1 quantitative reagent is peripheral blood. Peripheral blood is easy to obtain and is acceptable to people.

[0066] In some examples, the limb nerve entrapment injury is a tumor, especially a limb nerve entrapment injury caused by a benign tumor.

[0067] The second aspect of the present invention provides: A diagnostic or auxiliary diagnostic system for limb nerve entrapment injury, comprising: An LRG1 quantification device for quantifying the amount of LRG1 in a sample; An analysis device for determining the risk of peripheral nerve entrapment injury based on the LRG1 content; A result output device for outputting the analysis result of the analysis device.

[0068] There is no special requirement for the type of the LRG1 quantification device, as long as it can effectively quantify the amount of LRG1. In some examples, the LRG1 quantification device is an ELISA detection device, an immunoblotting device, an immunofluorescence device or a mass spectrometry device.

[0069] In some examples, a high LRG1 content indicates a high risk of peripheral nerve entrapment injury.

[0070] In some examples, a high LRG1 content means that its content in the peripheral blood sample is higher than 5 ng / mL.

[0071] In some examples, the sample is peripheral blood.

[0072] In some examples, the peripheral nerve entrapment injury is caused by a tumor, especially a benign tumor.

[0073] Screening and collection of clinical samples: Collect 30 cases of peripheral nerve entrapment injury caused by limb benign tumors in the past 5 years, and 30 cases of other upper limb benign masses without nerve entrapment. Record gender, age. All patients are measured for blood pressure, blood lipid, blood sugar, liver function, kidney function, four coagulation items, ECG, etc. All cases are confirmed by ultrasound, CT and nerve electrophysiology. At the same time, 60 normal people with matching age, gender and other conditions are selected as controls. Specimen collection: Collect 5 mL of venous blood from patients meeting the inclusion criteria, let it stand for 15 min, centrifuge at 3000 r / min for 15 min, extract the serum, put it into a sterile cryopreservation tube, and store it at -80 °C for research. Re-draw blood for detection in patients with nerve entrapment injury after surgery.

[0074] Study on blood samples of case patients Separate the serum. The inspection method is as follows: Under the action of a high-speed centrifuge, separate the serum and cells.

[0075] Main equipment, materials and reagents: homogenizer, high-speed centrifuge, 3 kDa ultrafiltration centrifugal tube, ICP-MS. The water used in the experiment process is ultrapure water with a resistivity of 18.2 MΩ / cm. Sample pretreatment methods: One full-automatic biochemical analyzer (one Beckman DXC800 and one AU400 each), one ten-thousandth electronic analytical balance (Shimadzu), microplate reader, Roche electrochemiluminescence automatic immunoassay analyzer (Elecsys2010), specific protein analyzer (imported), pure water machine (MEDICA60), electrothermal constant temperature water bath (DK-600), low-temperature high-speed centrifuge (sigma), ultra-low temperature freezer (Thermo Fisher Scientific, USA), low-temperature freezer (HFC350, Germany), etc. The above equipment conditions can fully ensure the implementation of this project.

[0076] Statistical methods: The T-test was used to compare the means of each group. The SPSS statistical software was used, and P < 0.05 was considered statistically significant.

[0077] Results: The normal value was determined as the mean plus 2 standard deviations as the normal range. The normal upper limit of the LRG1 detection kit produced in Huizhou (LRG1 Huizhou) was 5 ng / ml. The reagent produced in Huizhou was used to detect 30 cases of benign tumors in the limbs without nerve compression and 30 cases of benign tumors in the limbs with nerve compression. The CUTOFF value was 5 ng / ml. The results exceeded the CUTOFF in 28 cases, with 2 false negatives, and the sensitivity was 93%. There were 60 normal people, 12 of whom exceeded the CUTOFF value, with a false positive rate of 20% and a specificity of 80%.

[0078] The changes in peripheral blood LRG1 in patients with nerve compression injury of the limbs are shown in Table 1.

[0079] Table 1: Changes in peripheral blood LRG1 in patients with benign tumors of the limbs accompanied by nerve compression injury (X ± SD, unit: ng / ml) Number of people LRG1 (Huizhou) / ng / ml Normal people 60 3.2±1.1 Peripheral tumors without nerve compression injury 30 3.6±1.0 Peripheral tumors with nerve compression injury 30 12.6± 2.1 Compared with normal people and those without nerve compression, the level of LRG1 in patients with nerve compression injury of the limbs was significantly increased, P < 0.001. There was no difference between those with simple benign tumors without nerve compression injury and normal people.

[0080] Sensitivity and specificity of LRG1 in diagnosing nerve compression injury of the limbs Sensitivity = number of true positives / (number of true positives + number of false negatives) * 100%, the percentage of correctly diagnosing patients Specificity = number of true negatives / (number of true negatives + number of false positives) * 100%, the percentage of correctly diagnosing non-patients False positive rate = number of false positives / number of negative cases in the gold standard False negative rate = number of false negatives / number of positive cases in the gold standard The normal upper limit of the LRG1 detection kit produced in Huizhou is 5 ng / ml. The reagent produced in Huizhou was used to detect 30 cases of limb tumors with nerve compression injury. The CUTOFF value was 5 ng / ml. The results showed that 28 cases exceeded the CUTOFF value, 2 cases were false negatives, and the sensitivity reached 93.33%. Among 60 normal people, 12 exceeded the CUTOFF value, and the false positive rate was 20%.

[0081] The changes in LRG1 before and after the treatment of limb nerve compression injury are shown in Table 2.

[0082] Table 2: Changes in LRG1 before and after the treatment of limb nerve compression injury Number of cases LRG1 / ng / ml Normal people 60 3.2±1.1 Before surgery for peripheral tumors with nerve compression injury 30 12.6± 2.1 After surgery for peripheral tumors with nerve compression injury 19 3.2± 2.3 After the operation of limb nerve compression injury, LRG1 decreased significantly compared with that before the operation, P < 0.001, and was comparable to the level of normal people, P > 0.05.

[0083] The above test results show that when limb tumors cause nerve compression, the average value of LRG1 in the blood is higher than that of normal people, and it increases more significantly with the aggravation of the injury. As a peripheral blood marker for limb tumors causing nerve compression, it has high sensitivity and specificity.

[0084] The present invention first proposes and verifies that LRG1 is a peripheral blood marker for limb tumors causing nerve compression, and its results can be used for the diagnosis of limb tumors causing nerve compression.

[0085] The above is a further detailed description of the present invention, and it should not be regarded as a limitation to the specific implementation of the present invention. For those of ordinary skill in the technical field to which the present invention belongs, simple deductions or substitutions without departing from the concept of the present invention are within the protection scope of the present invention.

[0086] References: [1] Tessa Gordon 1, Nasim Amirjani, David C Edwards, K Ming Chan,Brief post-surgical electrical stimulation accelerates axon regeneration and muscle reinnervation without affecting the functional measures in carpal tunnel syndrome patients. Trial Exp Neurol. 2010 May;223(1):192 - 202. [2] Proteomic analysis of hippocampus in mice following long-term exposure to low levels of copper. Qian Sun, Ming Ying, Quan Ma, Zhijun Huang, Liangyu Zou, Jianjun Liu, Zhixiong Zhuang, Xifei Yang. Toxicol. Res., 2016, 4, 28 [3] Serum proteomic analysis reveals potential serum biomarkers for occupational medicamentosa-like dermatitis caused by trichloroethylene Peiwu Huang a,d,1,Xiaohu Ren a,1, Zhijun Huang c,1, Xifei Yang a, Wenxu Hong a,Yanfang Zhang b, Hang Zhang a,d, Wei Liu a, Haiyan Huang a, Xinfeng Huang a,Desheng Wu a,Linqing Yang a, Haiyan Tang b, Li Zhou a, Xuan Li a, Jianjun Liu. Toxicology Letters 2014, 6, 21(sci 3.75) [4] 1. Ditochondrial proteomic alterations caused by long-term low –dose copper exposure in mouse cortex. Xuemei Lin, Gang Wei, Zhijun Huang, etc. Toxicology Letter, 2016 [5]. Detection of the coagulation factor VII R353Q genotype in Han patients with intracerebral hemorrhage in South China. 2007, 6 《Chinese Journal of Arteriosclerosis》 [6]. Research progress on the application of gene chips in the diagnosis and treatment of cardiovascular diseases. 2005, 10 《China Medical Engineering》 [7]. Application of factor DNA molecular genetic markers RFLP, STR, SNP in coagulation genotype analysis. 2006, 4 《Chinese Journal of Misdiagnostics》。

Claims

1. Use of an LRG1 quantitative reagent in the preparation of a reagent for diagnosing or assisting in the diagnosis of limb nerve compression injury.

2. The application according to claim 1, characterized in that, The LRG1 quantitative reagent is an ELISA reagent, an immunofluorescence reagent, or a mass spectrometry analysis reagent.

3. The application according to claim 1, characterized in that The sample detected by the LRG1 quantitative reagent is peripheral blood.

4. The application according to claim 1, characterized in that, The limb nerve compression injury is a limb nerve compression injury caused by a tumor.

5. A diagnostic or auxiliary diagnostic system for limb nerve compression injury, comprising: an LRG1 quantification device for quantifying the amount of LRG1 in a sample; an analysis device for determining the risk of limb nerve compression injury based on the content of LRG1; a result output device for outputting the analysis result of the analysis device.

6. The diagnosis system for limb nerve entrapment injury according to claim 5, characterized in that, The LRG1 quantification device is an ELISA detection device, an immunoblotting device, an immunofluorescence device, or a mass spectrometry device.

7. The diagnosis system for limb nerve compression injury according to claim 5, wherein A high content of LRG1 indicates a high risk of limb nerve compression injury.

8. The diagnosis system for limb nerve compression injury according to claim 7, characterized in that, A high content of LRG1 means that its content in the peripheral blood sample is higher than 5 ng / mL.

9. The diagnosis system for limb nerve compression injury according to claim 5, wherein The sample is peripheral blood.

10. The diagnosis system for limb nerve entrapment injury according to claim 5, wherein The limb nerve compression injury is a limb nerve compression injury caused by a tumor.