Quantitative experiment research method for influence of semiconductor laser on functional activity of neural stem

Abstract

The invention provides two quantitative experiment research methods for influence of semiconductor laser on functional activity of a neural stem. The invention discloses a quantitative experiment research method for influence of semiconductor laser on functional activity of a nerve stem. The quantitative experiment research method sequentially comprises the following steps: (1) connection of a nerve stem specimen and an instrument: putting an in-vitro nerve stem into a multi-electrode nerve specimen box, wherein the adopted multi-electrode nerve specimen box comprises a pair of stimulating electrodes, a grounding electrode and two pairs of recording electrodes; (2) electrically stimulating the nerve trunk; (3) irradiating the nerve stem with laser: irradiating the local section of the nerve stem on the bearing platform by using a semiconductor laser source, and recording the irradiation time length, wavelength and irradiation intensity of the semiconductor laser; (4) electrically stimulating the nerve trunk again; and (5) data processing. According to the invention, the influence result of laser irradiation on the action potential waveform amplitude and conduction velocity of the neural stem can be quantitatively revealed.

Description

Technical Field

[0001] The present invention belongs to the technical field of body function experimental research, and particularly relates to a quantitative experimental research method for the influence of two semiconductor lasers on the functional activity of nerve trunks. Background Art

[0002] The nerve trunk is an important structure of the body's nervous system. It can generate and conduct action potentials, thereby realizing the rapid long-distance conduction of regulatory information in the body, which is crucial for the function and coordination of various systems and organs in the body. The nerve trunk compound action potentials (abbreviated as nerve trunk AP, where AP is the English abbreviation of action potentials), is a composite electrical signal recorded on the surface of the nerve trunk when many nerve fibers contained in the nerve trunk generate action potentials due to excitation and conduct to the recording electrode. It is also an important manifestation of the excitatory functional activity of the nerve trunk. Among them, the waveform amplitude and conduction velocity of the nerve trunk action potential are the main indicators of the excitatory functional activity of the nerve trunk.

[0003] In recent years, semiconductor lasers have gradually gained attention in biomedical research and have also begun to be clinically applied. For example, in research related to optogenetic technology, lasers with specific wavelengths emitted by semiconductor lasers can penetrate biological tissues and activate photosensitive proteins introduced into specific types of neurons through genetic engineering technology, thereby achieving precise stimulation and regulation of neurons; some studies have tried to directly irradiate the living nerves of animals within the surgical field of view with semiconductor lasers in order to change nerve function; again, in human physical therapy, especially in physical therapy for pain relief, semiconductor laser therapy devices are also common equipment, and its mechanism is also related to the fact that semiconductor lasers can change the excitatory function of local nerve trunks.

[0004] Theoretically, the influence of semiconductor lasers with different parameters (such as wavelength, intensity or power, irradiation time, etc.) on nerve trunk action potentials should be quantitatively studied, which is of great significance for deeply understanding the characteristics of semiconductor lasers in changing nerve trunk function and thus more effectively using lasers to improve nervous system function. However, in the prior art, no convenient, practical, scientific and reliable quantitative experimental research method for the influence of semiconductor lasers with different parameters (such as wavelength, intensity, irradiation time) on the waveform amplitude and conduction velocity of action potentials of isolated nerve trunks has been found. The internal reasons are as follows: The potential of the nerve trunk is actually very weak (about 10 mV) and the duration is short (1 - 2 ms), and any minor external changes may affect the result analysis. Moreover, the nerve trunk specimen is an in vitro biological specimen, which will gradually change with the extension of the in vitro time, and the biological properties of the nerve trunk specimen may also change with the increase in the number of electrical stimulations. In order to compare the changes in the functional activity of the in vitro nerve trunk before and after laser irradiation, the common idea that comes to mind is to first perform electrical stimulation on the in vitro nerve and measure the action potential of the nerve trunk before laser irradiation, then irradiate the in vitro nerve with laser, and then measure the action potential of the nerve trunk on the in vitro nerve after laser irradiation, and finally compare the action potentials of the nerve trunk before and after laser irradiation. If the action potentials of the two groups of nerve trunks obtained according to this common idea may be different, but the above differences may be caused by the following two other factors in addition to possibly being caused by laser irradiation: First, the time points for obtaining the action potential data of the two groups of nerve trunks are different; Second, the number of electrical stimulations experienced by the nerve trunk specimen when obtaining the action potential data of the two groups is different. Therefore, if the experimental research is carried out according to the above common idea, it is difficult to separate the changes caused solely by the laser irradiation factor from the multi-factor results, and it is difficult to quantitatively reveal the precise influence result of laser irradiation on the nerve trunk. Summary of the Invention

[0005] The first object of the present invention is to provide a quantitative experimental research method for the influence of semiconductor laser on the functional activity of nerve trunk, which can quantitatively reveal the influence result of laser irradiation on the waveform amplitude of the nerve trunk action potential.

[0006] The second object of the present invention is to provide a quantitative experimental research method for the influence of semiconductor laser on the functional activity of nerve trunk, which can quantitatively reveal the influence result of laser irradiation on the conduction velocity of the nerve trunk action potential.

[0007] The first object of the present invention can be achieved according to the following scheme: A quantitative experimental research method for the influence of semiconductor laser on the functional activity of nerve trunk, characterized by successively including the following steps: (1)Connection of the nerve trunk specimen and the instrument: Place the excised nerve trunk into a multi-electrode nerve specimen chamber. The multi-electrode nerve specimen chamber used includes a pair of stimulating electrodes, a grounding electrode, and two pairs of recording electrodes. The pair of stimulating electrodes consists of a first stimulating electrode and a second stimulating electrode. The first pair of recording electrodes consists of a first recording electrode and a second recording electrode. The second pair of recording electrodes consists of a third recording electrode and a fourth recording electrode. The seven electrodes, namely the first stimulating electrode, the second stimulating electrode, the grounding electrode, the first recording electrode, the second recording electrode, the third recording electrode, and the fourth recording electrode, are arranged in sequence along the horizontal direction. A supporting platform for supporting the nerve trunk is provided between the second recording electrode and the third recording electrode. The nerve trunk is placed on the first stimulating electrode, the second stimulating electrode, the grounding electrode, the first recording electrode, the second recording electrode, the supporting platform, the third recording electrode, and the fourth recording electrode. (2)Electrical stimulation of the nerve trunk: Apply electrical stimulation to the nerve trunk through the first stimulating electrode and the second stimulating electrode to induce the nerve trunk to generate action potentials and conduct them to the recording electrodes. Record the biphasic nerve trunk action potential waveform AP1 generated by the first pair of recording electrodes and the biphasic nerve trunk action potential waveform AP2 generated by the second pair of recording electrodes. (3)Laser irradiation of the nerve trunk: Use a semiconductor laser source to irradiate a local segment of the nerve trunk on the supporting platform, and record the irradiation time length, wavelength, and irradiation intensity of the semiconductor laser. (4)Re-electrical stimulation of the nerve trunk: Apply electrical stimulation to the nerve trunk again through the first stimulating electrode and the second stimulating electrode, and the parameters of the electrical stimulation are the same as those in step (2). Again, induce the nerve trunk to generate action potentials and conduct them to the recording electrodes. Record the biphasic nerve trunk action potential waveform AP3 generated by the first pair of recording electrodes and the biphasic nerve trunk action potential waveform AP4 generated by the second pair of recording electrodes. (5)Data processing: First, measure the maximum amplitude f of the upward waveform of the biphasic nerve trunk action potential AP1 1 , the maximum amplitude f of the upward waveform of the biphasic nerve trunk action potential AP2 2 , the maximum amplitude f of the upward waveform of the biphasic nerve trunk action potential AP3 3 , and the maximum amplitude f of the upward waveform of the biphasic nerve trunk action potential AP4 4 respectively. Finally, use the formula to calculate the amplitude influence degree value c = (f 4 / f 3 ) / (f 2 / f 1 ).

[0008] The above-mentioned amplitude influence degree value c reflects the influence degree of the semiconductor laser on the amplitude of the nerve trunk waveform under the laser parameter conditions in step (3). The closer the influence degree value c is to 1, the smaller the influence of the semiconductor laser under the laser parameter conditions on the amplitude of the nerve trunk action potential waveform; on the contrary, the farther the influence degree value c is from 1, the greater the influence of the semiconductor laser under the laser parameter conditions on the amplitude of the nerve trunk action potential waveform.

[0009] Furthermore, as long as the laser parameter conditions in step (3) are changed in series and the experimental steps are repeated, the amplitude influence degree values c under different laser wavelength / irradiation intensity / irradiation time length and other parameter conditions can be collected, and based on this, the relationship curves between the irradiation wavelength and the amplitude influence degree value c, the relationship curves between the irradiation intensity and the amplitude influence degree value c, and the relationship curves between the irradiation time length and the amplitude influence degree value c can be made, so as to very intuitively and quantitatively reveal the influence degree of various different wavelengths / irradiation intensities / irradiation time lengths, etc. of the semiconductor laser on the amplitude of the nerve trunk action potential waveform, as well as the change trend of this influence degree.

[0010] The second object of the present invention can be achieved according to the following scheme: A quantitative experimental research method for the influence of a semiconductor laser on the functional activity of a nerve trunk, characterized by successively including the following steps: (1) Connection of the nerve trunk specimen and the instrument: The excised nerve trunk is placed in a multi-electrode nerve specimen box. The multi-electrode nerve specimen box used includes a pair of stimulating electrodes, a grounding electrode, and two pairs of recording electrodes. A pair of stimulating electrodes consists of a first stimulating electrode and a second stimulating electrode. The first pair of recording electrodes consists of a first recording electrode and a second recording electrode. The second pair of recording electrodes consists of a third recording electrode and a fourth recording electrode. The seven electrodes of the first stimulating electrode, the second stimulating electrode, the grounding electrode, the first recording electrode, the second recording electrode, the third recording electrode, and the fourth recording electrode are arranged horizontally in sequence; A supporting platform for supporting the nerve trunk is provided between the second recording electrode and the third recording electrode; The nerve trunk is placed on the first stimulating electrode, the second stimulating electrode, the grounding electrode, the first recording electrode, the second recording electrode, the supporting platform, the third recording electrode, and the fourth recording electrode; (2) Electrical stimulation of the nerve trunk: Electrical stimulation is applied to the nerve trunk through the first stimulating electrode and the second stimulating electrode to induce the nerve trunk to generate an action potential and conduct it to the recording electrode; The biphasic nerve trunk action potential waveforms AP1 generated by the first pair of recording electrodes and the biphasic nerve trunk action potential waveforms AP2 generated by the second pair of recording electrodes are recorded; (3) Laser irradiation of the nerve trunk: The local segment of the nerve trunk on the supporting platform is irradiated by a semiconductor laser source, and the irradiation time length, wavelength, and irradiation intensity of the semiconductor laser are recorded; (4) Re - stimulate the nerve trunk again: Apply electrical stimulation to the nerve trunk again through the first stimulating electrode and the second stimulating electrode, and the parameters of the electrical stimulation are the same as those in step (2), and induce the nerve trunk to generate action potentials again and conduct them to the recording electrode; Record the biphasic nerve trunk action potential waveform AP3 generated by the first pair of recording electrodes and the biphasic nerve trunk action potential waveform AP4 generated by the second pair of recording electrodes; (5) Data processing: First, determine the time points when the upward waveform peaks of the biphasic nerve trunk action potential waveform AP1, the biphasic nerve trunk action potential waveform AP2, the biphasic nerve trunk action potential waveform AP3, and the biphasic nerve trunk action potential waveform AP4 appear; Then, measure the time interval length t between the time point when the electrical stimulation starts in step (2) and the time point when the waveform peak of the biphasic nerve trunk action potential waveform AP1 appears 1 , measure the time interval length t between the time point when the electrical stimulation starts in step (2) and the time point when the waveform peak of the biphasic nerve trunk action potential waveform AP2 appears 2 , measure the time interval length t between the time point when the electrical stimulation starts in step (4) and the time point when the waveform peak of the biphasic nerve trunk action potential waveform AP3 appears 3 , measure the time interval length t between the time point when the electrical stimulation starts in step (4) and the time point when the waveform peak of the biphasic nerve trunk action potential waveform AP4 appears 4 ; According to the formula v 1 = m / t 1 Calculate the average conduction velocity v of the nerve trunk action potential AP1 1 , according to the formula v 2 = n / t 2 Calculate the average conduction velocity v of the nerve trunk action potential AP2 2 , according to the formula v 3 = m / t 3 Calculate the average conduction velocity v of the nerve trunk action potential AP3 3 , according to the formula v 4 = n / t 4 Calculate the average conduction velocity v of the nerve trunk action potential AP4 4 , where m is the horizontal distance between the mid - point of the second stimulating electrode and the first pair of recording electrodes, n is the horizontal distance between the mid - point of the second stimulating electrode and the second pair of recording electrodes, and finally use the formula d=(v 4 / v 3 ) / ( v 2 / v 1 ) to calculate the conduction velocity influence degree value d.

[0011] The above influence degree value d reflects the influence degree of the semiconductor laser on the conduction velocity of the nerve trunk under the laser parameter conditions in step (3). The closer the conduction velocity influence degree value d is to 1, the smaller the influence of the semiconductor laser under the laser parameter conditions on the conduction velocity of the nerve trunk action potential; conversely, the farther the conduction velocity influence degree value d is from 1, the greater the influence of the semiconductor laser under the laser parameter conditions on the conduction velocity of the nerve trunk action potential; if the conduction velocity influence degree value d > 1, it indicates that the conduction velocity increases after laser irradiation, and vice versa, it decreases.

[0012] Furthermore, as long as the laser parameter conditions in step (3) are serially changed and the experimental steps (2) to (4) are repeated, the conduction velocity influence degree values d under different parameter conditions such as laser wavelength / irradiation intensity / irradiation time length can be collected, and based on this, the relationship curves between the irradiation wavelength and the conduction velocity influence degree value d, the relationship curves between the irradiation intensity and the conduction velocity influence degree value d, and the relationship curves between the irradiation time length and the conduction velocity influence degree value d can be plotted, so as to very intuitively and quantitatively reveal the influence degree of various parameters such as different wavelengths / irradiation intensities / irradiation time lengths of the semiconductor laser on the conduction velocity of the nerve trunk action potential, as well as the change trend of this influence degree.

[0013] The present invention has the following advantages and effects: First, in the experimental design scheme of the present invention, the electrical stimulation acts on the left end of the nerve trunk, and on the right side are successively the grounding electrode, the first pair of recording electrodes, the supporting platform, and the second pair of recording electrodes. In this case, the nerve trunk action potential conducts from left to right. The laser only irradiates a small section of the nerve trunk on the supporting platform between the first pair of recording electrodes and the second pair of recording electrodes. Therefore, the action potential of the first pair of recording electrodes is not affected by laser irradiation, while the action potential of the second pair of recording electrodes will be affected by laser irradiation; on the other hand, during the implementation of steps (2) to (4), the biological property changes of the entire nerve trunk due to non-laser factors such as time lapse and electrical stimulation are basically the same. Therefore, the change amplitude caused by non-laser factors of the second pair of recording electrodes can be estimated by calculating the change amplitude of the first pair of recording electrodes, that is, the action potential of the first pair of recording electrodes can be used as an autologous parallel control for the laser to affect the nerve trunk action potential. Furthermore, by using the action potential data of the first pair of recording electrodes, the influence of non-laser factors in the action potential data of the second pair of recording electrodes is eliminated, and finally, the influence of the laser irradiation factor is measured by using the action potential of the second pair of recording electrodes with reference to the action potential of the first pair of recording electrodes.

[0014] Second, specifically, the first solution of the present invention introduces the biphasic nerve trunk AP between the first pair of recording electrodes as a reference for the biphasic nerve trunk AP of the second pair of recording electrodes. When comparing the waveform amplitudes of the biphasic nerve trunk APs of the second pair of recording electrodes before and after laser irradiation (i.e., comparing f 4 and f 2 ), the factors of the waveform amplitudes (f 3 and f 1 ) of the biphasic nerve trunk APs of the first pair of recording electrodes before and after laser irradiation are introduced as references. Thus, the influences of "the acquisition time points of the two groups of nerve trunk action potential data are different" and "the number of electrical stimulation times experienced by the nerve trunk specimen during the acquisition of the two groups of nerve trunk action potential data" are basically eliminated (because the acquisition time points of the biphasic nerve trunk AP1 and the biphasic nerve trunk AP2 are basically the same, and the number of electrical stimulation times experienced by the nerve trunk specimen during acquisition is the same, and the acquisition time points of the biphasic nerve trunk AP3 and the biphasic nerve trunk AP4 are basically the same, and the number of electrical stimulation times experienced by the nerve trunk specimen during acquisition is the same). As a result, the influence of laser irradiation on the waveform amplitude of the nerve trunk action potential can be more purely stripped and quantitatively revealed. Furthermore, the relationship curves between the irradiation wavelength and the amplitude influence degree value c, the relationship curves between the irradiation intensity and the amplitude influence degree value c, and the relationship curves between the irradiation time length and the amplitude influence degree value c can be made, so as to very intuitively and quantitatively show the influence degree of semiconductor laser on the waveform amplitude of the nerve trunk action potential under different wavelengths / different intensities (powers) / different irradiation time lengths and other conditions, as well as the change trend of this influence degree.

[0015] Third, specifically, the second solution of the present invention introduces the biphasic nerve trunk AP between the first pair of recording electrodes as a reference for the biphasic nerve trunk AP of the second pair of recording electrodes. When comparing the conduction velocities of the biphasic nerve trunk APs of the second pair of recording electrodes before and after laser irradiation (comparing v 4 and v 2 ), the conduction velocities (v 3 and v 1Using the factors as a reference, the influences of "the acquisition time points of the action potential data of the two groups of nerve trunks are different" and "the number of electrical stimulation times experienced by the nerve trunk specimens when acquiring the action potential data of the two groups of nerve trunks are different" can be basically eliminated (because the acquisition time points of the biphasic nerve trunk AP1 and the biphasic nerve trunk AP2 are basically the same, and the number of electrical stimulation times experienced by the nerve trunk specimens when acquiring them is the same; the acquisition time points of the biphasic nerve trunk AP3 and the biphasic nerve trunk AP4 are basically the same, and the number of electrical stimulation times experienced by the nerve trunk specimens when acquiring them is the same). Thus, the influence result of laser irradiation on the conduction velocity of the nerve trunk action potential can be purely separated and quantitatively revealed. Furthermore, the relationship curves between the irradiation wavelength and the influence degree value d of the conduction velocity, the relationship curves between the irradiation intensity and the influence degree value d of the conduction velocity, and the relationship curves between the irradiation time length and the influence degree value d of the conduction velocity can be plotted, so as to very intuitively and quantitatively show the influence degree of semiconductor lasers on the conduction velocity of the nerve trunk action potential under different wavelengths / different intensities (powers) / different irradiation time lengths and other conditions, as well as the change trend of this influence degree. Description of the Drawings

[0016] Figure 1 It is a schematic diagram of a nerve trunk placed on a multi-electrode nerve specimen box.

[0017] Figure 2 It is a schematic diagram of the structure between a semiconductor laser source and a supporting platform.

[0018] Figure 3 It is a schematic diagram of the biphasic nerve trunk AP1 between the first recording electrode and the second recording electrode obtained in step (2).

[0019] Figure 4 It is a schematic diagram of the biphasic nerve trunk AP2 between the third recording electrode and the fourth recording electrode obtained in step (2).

[0020] Figure 5 It is a schematic diagram of the biphasic nerve trunk AP3 between the first recording electrode and the second recording electrode obtained in step (4).

[0021] Figure 6 It is a schematic diagram of the biphasic nerve trunk AP4 between the third recording electrode and the fourth recording electrode obtained in step (4). Detailed Implementation Modes Example 1

[0022] A quantitative experimental research method for the influence of semiconductor lasers on the functional activity of nerve trunks successively includes the following steps: (1) Connection of the nerve trunk specimen and the instrument: Place the active and good frog sciatic nerve trunk 8 into the multi-electrode nerve specimen box 10; Figure 1As shown, the multi - electrode nerve specimen box 10 adopted includes a pair of stimulating electrodes, a grounding electrode 5 and two pairs of recording electrodes. A pair of stimulating electrodes consists of a first stimulating electrode 6 and a second stimulating electrode 7. The first pair of recording electrodes consists of a first recording electrode 1 and a second recording electrode 2. The second pair of recording electrodes consists of a third recording electrode 3 and a fourth recording electrode 4. The first stimulating electrode 6, the second stimulating electrode 7, the grounding electrode 5, the first recording electrode 1, the second recording electrode 2, the third recording electrode 3, and the fourth recording electrode 4 are arranged in sequence horizontally from left to right. A supporting platform 9 for supporting the nerve trunk is provided between the second recording electrode 2 and the third recording electrode 3; the isolated frog sciatic nerve trunk 8 is placed on the first stimulating electrode 6, the second stimulating electrode 7, the grounding electrode 5, the first recording electrode 1, the second recording electrode 2, the supporting platform 9, the third recording electrode 3, and the fourth recording electrode 4. A small piece of filter paper is padded on the supporting platform 9 and soaked with Ringer's solution to keep the surface of the nerve trunk 8 moist with Ringer's solution; (2)Electrical stimulation of the nerve trunk: Apply electrical stimulation (in this embodiment, the electrical stimulation is a square wave with an intensity of 2 V and a pulse width of 0.05 ms) to the isolated frog sciatic nerve trunk 8 through the first stimulating electrode 6 and the second stimulating electrode 7, inducing the isolated frog sciatic nerve trunk 8 to generate action potentials and conduct them to the recording electrodes; Record the biphasic nerve trunk action potential waveform AP1 (as shown in Figure 3 ) generated by the first pair of recording electrodes (the first recording electrode 1 and the second recording electrode 2) and the biphasic nerve trunk action potential waveform AP2 (as shown in Figure 4 ) generated by the second pair of recording electrodes (the third recording electrode 3 and the fourth recording electrode 4): (3)Laser irradiation of the nerve trunk: Use a semiconductor laser source 12 to irradiate a local segment of the isolated frog sciatic nerve trunk 8 on the supporting platform 9, and record the irradiation time length, wavelength, and irradiation intensity of the semiconductor laser; A baffle 11 with a light - limiting aperture 110 is placed between the semiconductor laser source 12 and the supporting platform 9, as shown in Figure 2 ; In the first embodiment, the distance between the baffle 11 and the laser action platform 9 is 3 cm, the diameter of the light - limiting aperture is 0.3 cm. When laser irradiation is carried out, it passes through the light - limiting aperture 110 of the baffle 11, so that the influence of the laser is only limited to a small segment of the nerve placed on the laser action platform, and at the same time, it is convenient to carry out some related experimental operations (such as dripping Ringer's solution on the specimen during the experimental interval); (4)Re - electrical stimulation of the nerve trunk: After laser irradiation, apply electrical stimulation to the isolated frog sciatic nerve trunk 8 again through the first stimulating electrode 6 and the second stimulating electrode 7, and the parameters of the electrical stimulation are the same as those in step (2), inducing the isolated frog sciatic nerve trunk 8 to generate action potentials again and conduct them to the recording electrodes; Record the biphasic nerve trunk action potential waveform AP3 (as shown in Figure 5as shown) and the biphasic nerve trunk action potential waveform AP4 generated by the second pair of recording electrodes (as Figure 6 shown); 5) Data processing: First, measure the maximum amplitude of the upward waveform of the biphasic nerve trunk action potential AP1 ( Figure 3 f in 1 ), the maximum amplitude of the upward waveform of the biphasic nerve trunk action potential AP2 ( Figure 4 f in 2 ), the maximum amplitude of the upward waveform of the biphasic nerve trunk action potential AP3 ( Figure 5 f in 3 ), and the maximum amplitude of the upward waveform of the biphasic nerve trunk action potential AP4 ( Figure 6 f in 4 ); Finally, calculate the amplitude influence degree value c, c = (f 4 / f 3 / (f 2 / f 1 ).

[0023] In the above-mentioned first embodiment, multiple lasers with different wavelengths can be further selected, and for each wavelength, the experimental steps can be repeated according to the above method, and the amplitude influence degree value c under the irradiation conditions of lasers with different wavelengths can be measured, and based on this, a relationship curve between the wavelength and the influence degree value c can be made. Similarly, multiple lasers with different irradiation intensities / irradiation time lengths can be further selected and the experimental steps can be repeated according to the above method, and the amplitude influence degree value c under different irradiation intensities / irradiation time lengths can be measured, and based on this, a relationship curve between the irradiation intensity and the amplitude influence degree value c and a relationship curve between the irradiation time length and the amplitude influence degree value c can be made. By the same token, the irradiation distance and irradiation range of the laser can be changed, and the change trend of the amplitude influence degree value c under different irradiation distances / irradiation ranges can be further measured and studied. Example Two

[0024] The quantitative experimental research method for the influence of the semiconductor laser in Example Two on the functional activity of the nerve trunk includes steps (1) to (5) in sequence, where steps (1) to (4) in Example Two are the same as steps (1) to (4) in Example One and will not be elaborated; And step (5) in Example Two is as follows: Data processing: Determine the time point when the waveform peak of the biphasic nerve trunk action potential waveform AP1 appears ( Figure 3 T point in 1 ), the time point when the waveform peak of the biphasic nerve trunk action potential waveform AP2 appears ( Figure 4 T point in 2 ), the time point when the waveform peak of the biphasic nerve trunk action potential waveform AP3 appears ( Figure 5T in 3 point), the time point when the waveform peak of the biphasic compound nerve action potential AP4 appears ( Figure 5 T in 4 point); Next, measure the time point T when the electrical stimulation starts in step (2) 20 and the time point T when the waveform peak of the biphasic compound nerve action potential AP1 appears 1 The time interval length t between the two 1 , measure the time point T when the electrical stimulation starts in step (2) 20 and the time point T when the waveform peak of the biphasic compound nerve action potential AP2 appears 2 The time interval length t between the two 2 , measure the time point T when the electrical stimulation starts in step (4) 40 and the time point T when the waveform peak of the biphasic compound nerve action potential AP3 appears 3 The time interval length t between the two 3 , measure the time point T when the electrical stimulation starts in step (4) 40 and the time point T when the waveform peak of the biphasic compound nerve action potential AP4 appears 4 The time interval length t between the two 4 , according to the formula v 1 =m / t 1 Calculate the average conduction velocity v of the compound nerve action potential AP1 1 , according to the formula v 2 =n / t 2 Calculate the average conduction velocity v of the compound nerve action potential AP2 2 , according to the formula v 3 =m / t 3 Calculate the average conduction velocity v of the compound nerve action potential AP3 3 , according to the formula v 4 =n / t 4 Calculate the average conduction velocity v of the compound nerve action potential AP4 4 , where m is the lateral distance between the midpoint of the second stimulating electrode 7 and the first pair of recording electrodes (i.e., the lateral midpoint between the first recording electrode 1 and the second recording electrode 2), and n is the lateral distance between the midpoint of the second stimulating electrode 7 and the second pair of recording electrodes (i.e., the lateral midpoint between the third recording electrode 3 and the fourth recording electrode 4), as Figure 1 shown; Finally, calculate the conduction velocity influence degree value d, d=( v 4 / v 3 ) / ( v 2 / v 1 )。

[0025] In the second embodiment described above, multiple lasers with different wavelengths can be further selected. By repeating the experimental steps for each wavelength according to the above method, the influence degree value d of the conduction velocity under the irradiation of lasers with different wavelengths can be measured, and based on this, a relationship curve between the wavelength and the influence degree value d of the conduction velocity can be plotted. Similarly, multiple lasers with different irradiation intensities / irradiation time lengths can be further selected to repeat the experimental steps, and the influence degree value d of the conduction velocity under different irradiation intensities / irradiation time lengths can be measured, and based on this, a relationship curve between the irradiation intensity and the influence degree value d of the conduction velocity and a relationship curve between the irradiation time length and the influence degree value d of the conduction velocity can be plotted. By the same token, the irradiation distance and irradiation range of the laser can be changed, and the change trend of the influence degree value d of the conduction velocity under different irradiation distances / irradiation ranges can be further measured and studied.

Claims

1. A quantitative experimental research method for the effect of semiconductor laser on the functional activity of nerve stems, characterized in that The following steps are included in sequence: (1) Connection between nerve trunk specimen and instrument: Place the isolated nerve trunk into a multi-electrode nerve specimen box, the multi-electrode nerve specimen box comprising a pair of stimulating electrodes, a grounding electrode and two pairs of recording electrodes, the pair of stimulating electrodes comprising a first stimulating electrode and a second stimulating electrode, the first pair of recording electrodes comprising a first recording electrode and a second recording electrode, and the second pair of recording electrodes comprising a third recording electrode and a fourth recording electrode; a total of seven electrodes, namely, the first stimulating electrode, the second stimulating electrode, the grounding electrode, the first recording electrode, the second recording electrode, the third recording electrode and the fourth recording electrode, are sequentially arranged in a transverse direction, a supporting platform for supporting the nerve trunk is provided between the second recording electrode and the third recording electrode; the nerve trunk is placed on the first stimulating electrode, the second stimulating electrode, the grounding electrode, the first recording electrode, the second recording electrode, the supporting platform, the third recording electrode and the fourth recording electrode; (2) Electrical stimulation of the nerve trunk: applying electrical stimulation to the nerve trunk through the first stimulation electrode and the second stimulation electrode to induce the nerve trunk to generate action potentials and conduct them to the recording electrode; recording the biphasic nerve trunk action potential waveform AP1 generated by the first pair of recording electrodes and the biphasic nerve trunk action potential waveform AP2 generated by the second pair of recording electrodes; (3) Laser irradiation of nerve trunk: Use a semiconductor laser source to irradiate a local segment of the nerve trunk on the supporting platform, and record the irradiation time, wavelength, and intensity of the semiconductor laser; (4) Electrically stimulating the nerve trunk again: applying electrical stimulation to the nerve trunk again through the first stimulation electrode and the second stimulation electrode, and the parameters of the electrical stimulation are the same as those in step (2), again inducing the nerve trunk to generate an action potential and conduct it to the recording electrode; recording the biphasic nerve trunk action potential waveform AP3 generated by the first pair of recording electrodes and the biphasic nerve trunk action potential waveform AP4 generated by the second pair of recording electrodes; (5) Data processing: First, measure the maximum amplitude f1 of the upward waveform of the biphasic nerve trunk action potential AP1, the maximum amplitude f2 of the upward waveform of the biphasic nerve trunk action potential AP2, the maximum amplitude f3 of the upward waveform of the biphasic nerve trunk action potential AP3, and the maximum amplitude f4 of the upward waveform of the biphasic nerve trunk action potential AP4; finally, use the formula c=(f4 / f3) / (f2 / f1) to calculate the amplitude influence value c.

2. A quantitative experimental research method for the effect of semiconductor laser on the functional activity of nerve stems, characterized in that The following steps are included in sequence: (1) Connection between nerve trunk specimen and instrument: Place the isolated nerve trunk into a multi-electrode nerve specimen box, which includes a pair of stimulating electrodes, a grounding electrode and two pairs of recording electrodes. The pair of stimulating electrodes consists of a first stimulating electrode and a second stimulating electrode. The first pair of recording electrodes consists of a first recording electrode and a second recording electrode. The second pair of recording electrodes consists of a third recording electrode and a fourth recording electrode. The first stimulating electrode, the second stimulating electrode, the grounding electrode, the first recording electrode, the second recording electrode, the third recording electrode and the fourth recording electrode are arranged in a horizontal arrangement in sequence. A supporting platform for supporting the nerve trunk is provided between the second recording electrode and the third recording electrode. The nerve trunk is placed on the first stimulating electrode, the second stimulating electrode, the grounding electrode, the first recording electrode, the second recording electrode, the supporting platform, the third recording electrode and the fourth recording electrode. (2) Electrical stimulation of the nerve trunk: applying electrical stimulation to the nerve trunk through the first stimulation electrode and the second stimulation electrode to induce the nerve trunk to generate action potentials and conduct them to the recording electrode; recording the biphasic nerve trunk action potential waveform AP1 generated by the first pair of recording electrodes and the biphasic nerve trunk action potential waveform AP2 generated by the second pair of recording electrodes; (3) Laser irradiation of nerve trunk: Use a semiconductor laser source to irradiate a local segment of the nerve trunk on the supporting platform, and record the irradiation time, wavelength, and intensity of the semiconductor laser; (4) Electrically stimulating the nerve trunk again: applying electrical stimulation to the nerve trunk again through the first stimulation electrode and the second stimulation electrode, and the parameters of the electrical stimulation are the same as those in step (2), again inducing the nerve trunk to generate an action potential and conduct it to the recording electrode; recording the biphasic nerve trunk action potential waveform AP3 generated by the first pair of recording electrodes and the biphasic nerve trunk action potential waveform AP4 generated by the second pair of recording electrodes; (5) Data processing: first determine the time points at which the upward waveform peaks appear in the biphasic nerve trunk action potential waveforms AP1, AP2, AP3, and AP4; then, measure the time interval t1 between the time point at which the electrical stimulation starts in step (2) and the time point at which the waveform peak appears in the biphasic nerve trunk action potential waveform AP1, measure the time interval t2 between the time point at which the electrical stimulation starts in step (2) and the time point at which the waveform peak appears in the biphasic nerve trunk action potential waveform AP2, and measure the time interval t3 between the time point at which the electrical stimulation starts in step (4) and the time point at which the waveform peak appears in the biphasic nerve trunk action potential waveform AP3. The time interval t3 is measured in step (4) between the time when the electrical stimulation is started and the time when the peak of the biphasic nerve trunk action potential waveform AP4 appears; the average conduction velocity v1 of the nerve trunk action potential AP1 is calculated according to the formula v1=m / t1, the average conduction velocity v2 of the nerve trunk action potential AP2 is calculated according to the formula v2=n / t2, the average conduction velocity v3 of the nerve trunk action potential AP3 is calculated according to the formula v3=m / t3, and the average conduction velocity v4 of the nerve trunk action potential AP4 is calculated according to the formula v4=n / t4, where m is the lateral distance between the second stimulation electrode and the midpoint of the first pair of recording electrodes, and n is the lateral distance between the second stimulation electrode and the midpoint of the second pair of recording electrodes. Finally, the conduction velocity influence value d is calculated using the formula d=(v4 / v3) / (v2 / v1).

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