Application of pulse ultrasound instrument in preparation of medical instrument for treating and / or preventing diabetic cataract of subject

Abstract

The invention belongs to the technical field of biology, and particularly relates to application of a pulse ultrasound instrument in preparation of a medical instrument for treating and / or preventing diabetic cataract of a subject. It is found for the first time that the expression level of epithelial marker protein E-cadherin can be improved, the expression level of interstitial marker protein alpha-SMA can be reduced, the development process of the diabetic cataract can be delayed, and the morbidity of the diabetic cataract can be reduced by irradiating the liver through a pulse ultrasonic instrument.

Description

Technical Field

[0001] The present application relates to the field of biotechnology, and in particular to the use of a pulsed ultrasound apparatus in the preparation of a medical device for treating and / or preventing diabetic cataracts in a subject. Background Art

[0002] Diabetic cataract (DC) is one of the complications of diabetes. It is caused by increased blood sugar concentration, which leads to increased osmotic pressure of aqueous humor in the eye, disordered glucose metabolism in the lens, which causes the lens to absorb water and swell, and then break or disintegrate, and finally the lens becomes completely turbid, causing blurred vision. The main symptoms of DC are simultaneous onset of both eyes, decreased vision, rapid progression, and snowflake-like turbidity of the lens. At present, the incidence of cataracts in diabetic patients is increasing year by year, and about 60%-65% of diabetic patients are accompanied by DC. The main characteristics of DC are rapid progression and rapid maturation of lens opacity. Epidemiological studies have shown that the prevalence of DC is 5 times higher than that of non-diabetic cataracts, and the age of onset is 20 years earlier than that of non-diabetic cataracts. The high incidence and high blindness rate of DC make it a serious social problem.

[0003] Unstable glucose levels in the body lead to changes in the morphology and function of the lens, especially the lens epithelial cells are most severely damaged. Although the lens is not vascularized and provides nutrients and oxygen through the aqueous humor, the glucose concentration in the blood will still affect the glucose content in the aqueous humor. Therefore, hyperglycemia is considered to be a key factor that promotes lens changes in diabetic patients. Of course, as a metabolic disease, there are also changes in various metabolic substances in the body of diabetic patients, which also affect the occurrence and development of cataracts. The specific pathogenesis of DC is still controversial and needs further study.

[0004] Currently, the main method for treating DC is surgical removal of cataracts and artificial lens implantation. Although surgical treatment has the characteristics of small incisions and short time. However, compared with non-diabetic patients, diabetic patients have a higher risk of intraoperative and postoperative complications, which can easily lead to inflammation and eye tissue damage in patients. In addition, many studies have introduced metabolic-related substances to treat DC through drug administration, but the difference between the substances introduced from the outside and the substances produced by the body has made its efficacy very limited. Summary of the invention

[0005] In view of the above-mentioned shortcomings of the prior art, the present invention provides the use of a pulsed ultrasound device in the preparation of a medical device for treating and / or preventing DC, so as to solve the above-mentioned technical problems such as the high risk of intraoperative and postoperative complications in patients, inflammation and eye tissue damage in patients, the gap between externally introduced substances and the body's own production of substances, and minimal therapeutic effect.

[0006] To achieve the above objectives, the scheme of this application is as follows:

[0007] In a first aspect, the present invention provides the use of a pulsed ultrasound apparatus in the preparation of a medical device for treating and / or preventing diabetic cataracts in a subject.

[0008] Optionally, the pulsed ultrasound device is used to output 180-220 mW / cm 2 The average sound intensity.

[0009] Optionally, the average sound intensity is output in a pulse mode.

[0010] Optionally, the pulsed ultrasound machine is used to irradiate the subject's liver.

[0011] Optionally, the irradiation frequency is 8-12 min / day for at least 30 consecutive days.

[0012] Optionally, during the irradiation process, the duty cycle is 15%-25%, and the ultrasonic frequency is 280-300kHz.

[0013] In a second aspect, the present invention provides the use of a pulsed ultrasound apparatus in the preparation of a medical device for increasing the expression level of the epithelial marker protein E-cadherin in a subject.

[0014] In a third aspect, the present invention provides the use of a pulsed ultrasound apparatus in the preparation of a medical device for reducing the expression level of interstitial marker protein α-SMA in a subject.

[0015] In a fourth aspect, the present invention provides the use of a pulsed ultrasound apparatus in the preparation of a medical device for delaying the progression of diabetic cataracts in a subject.

[0016] In a fifth aspect, the present invention provides the use of a pulsed ultrasound apparatus in the preparation of a medical device for reducing the incidence of diabetic cataracts in a subject.

[0017] Beneficial effects of the present invention:

[0018] This application discovered for the first time that irradiating the liver of a subject using a pulsed ultrasound device can increase the expression level of the epithelial marker protein E-cadherin, reduce the expression level of the interstitial marker protein α-SMA, delay the progression of diabetic cataracts, and reduce the incidence of diabetic cataracts. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the structures shown in these drawings without paying creative work.

[0020] Figure 1 This is the experimental flow chart of this application;

[0021] Figure 2 2A is a graph showing the test results of blood glucose related indicators, 2B is a graph showing the test results of fasting blood glucose (the ordinate is the concentration of circulating blood glucose), 2C is a graph showing the test results of body weight (the ordinate is body weight), 2D is a graph showing the test results of oral glucose tolerance test (the ordinate is the blood glucose concentration), 2E is a graph showing the test results of AUC (i.e., area under the curve, AUC = (1 / 2*fasting blood glucose + 1h + 1 / 2*2h) * 1h), 2F is a graph showing the test results of fasting serum insulin level (the ordinate is the fasting serum insulin concentration), 2G is a graph showing the test results of HOMA (HOMA = fasting insulin (μU / mL) × Fasting glucose (mmol / L) / 22.5),

[0022] Figure 3 Figure 3A shows the incidence and severity of cataracts. 3A shows the cataract screening record (slit lamp microscope, 40×) (Grand 0: transparent; Grand 1: peripheral vesicles; Grand 2: vesicles and cortical opacities; Grand 3: diffuse central opacity; Grand 4: nuclear cataracts. 3B shows the number and grade of lenses with DC in rats of each group at the same time point (the ordinate is the number of lenses with DC of different opacities). 3C shows the incidence of rats in each group at the 2nd, 4th, 6th, 8th, and 10th week after the start of treatment (opacity index = number of eyeballs of each grade × cataract grade / total number of eyeballs) (the ordinate is the incidence of DC). 3D shows the ocular appearance of the LIPUS group and the model group (Model group) after the end of treatment.

[0023] Figure 4Figures 4A and 4B are the results of pathological analysis of the anterior capsule and equatorial part of the lens tissue by hematoxylin-eosin (H&E) staining, immunoblotting and real-time quantitative polymerase chain reaction (qRT-PCR). Figure 4A is the result of H&E staining of rat lens tissue ((anterior capsule: upper figure, equatorial part: lower figure)), and the black arrows indicate liquefied vacuoles. Figures 4B and 4C are the results of Western Blot analysis of the expression of E-cadherin and α-SMA proteins in lens tissue, respectively. Figures 4D and 4E are the results of qRT-PCR analysis of the expression of E-cadherin and α-SMA mRNA in lens tissue, respectively.

[0024] Figure 5 This is a graph showing the results of blood biochemical analysis. DETAILED DESCRIPTION

[0025] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0026] In the following description, numerous details are discussed to provide a more thorough explanation of embodiments of the present invention; however, it is apparent to one skilled in the art that embodiments of the present invention may be practiced without these specific details.

[0027] (I) Construction and grouping of rat T2DM model

[0028] Experimental animals: 120 4-week-old SPF-grade male Sprague-Dawley (SD) rats were purchased from the Experimental Animal Center of Chongqing Medical University;

[0029] like Figure 1 As shown, a rat T2DM model was constructed, and the specific steps were as follows:

[0030] Four-week-old male SD rats were adaptively housed for 1 week and provided with drinking water ad libitum in a housing environment with a temperature of (25±3)°C and a relative humidity of (40±10)%, with a 12-h day and night cycle, and free access to food and water;

[0031] After the start of the experiment, the animals were maintained on a high-fat diet (D12451) (45% calories as fat) for 4 weeks to induce obesity and insulin resistance in SD rats;

[0032] After 4 weeks of high-fat diet, 150 rats (≥300 g) were randomly divided into a normal group (Normal, n=30), a model group (Model, n=30), an ultrasound group (LIPUS, n=30), a metformin treatment group (MET, n=30) and a metformin combined with ultrasound treatment group (LIPUS+MET, n=30). After fasting for 16 h, the rats in the Model group, LIPUS group, MET group and LIPUS+MET group were intraperitoneally injected with a low dose of streptozotocin (STZ, 18883-66-4, Solarbio, Beijing Solaibao Technology Co., Ltd, Beijing, China) (35 mg.kg -1 , 0.1M citric acid buffer, pH 4.5ip), was used to induce moderate levels of islet cell death, impair insulin secretion, and make rats form a model similar to late T2DM; the normal group (Normal group) rats were intraperitoneally injected with the same concentration of citric acid buffer at the same time as the modeling rats (the injection dose depends on the weight of the rats);

[0033] The blood glucose changes of modeling mice were recorded by a handheld blood glucose meter (Sinocare GA-3, Sinocare Inc., Changsha, China) at 48h, 72h and 96h after STZ injection. Rats with blood glucose levels (i.e., circulating blood glucose) ≥16.7mmol / L for 7d (i.e., 7 days) were considered to have successfully established a diabetes model.

[0034] All experimental procedures were performed in accordance with the guidelines of the Ethics Committee of Chongqing Medical University to ensure compliance with ethical and scientific standards;

[0035] Both the MET group and the LIPUS+MET group were given MET (i.e., metformin, B1970, Apexbio, Shanghai Weihuan Biotechnology Co., Ltd, Shanghai, China) drug, and the physiological saline mixture was prepared. All formulations were kept at pH = 7.2 to prevent physiological pain or gastrointestinal discomfort. The formulations were prepared daily to prevent degradation caused by storage of the drug aqueous solution. For consistency, the concentration was mg.kg -1 The difference between the MET group and LIPUS+MET is that the latter was treated with LIPUS stimulation. The irradiation parameters of LIPUS stimulation were: ultrasound intensity 200 mW / cm 2 , irradiation time 10min / d, duration 30d, duty cycle 20%, ultrasonic frequency 299kHz, modulation frequency 1kHz, working mode is pulse mode;

[0036] After successful modeling in the ultrasound group (LIPUS, n = 30), LIPUS stimulation was applied, and no MET drug was given, that is, the ultrasound group only received LIPUS stimulation;

[0037] After the model was successfully established, the normal group (Normal, n = 30) did not undergo LIPUS stimulation and did not receive MET drugs. That is, the normal group did not undergo LIPUS stimulation and did not receive MET drugs.

[0038] After the model was successfully established, the model group (Model, n = 30) was subjected to LIPUS sham stimulation (LIPUS without actual energy output). The sham stimulation time was the same as that of the LIPUS group, that is, the model group was not given MET drugs, and only LIPUS sham stimulation (LIPUS without actual energy output) was performed;

[0039] The dosage concentration of metformin was 200 mg / kg / d, and metformin was administered by gavage before ultrasound stimulation or sham ultrasound stimulation;

[0040] After the corresponding groups of rats were administered, they were anesthetized and subjected to LIPUS stimulation or sham stimulation, specifically:

[0041] All rats were anesthetized with 1–4% isoflurane (RWD, R510-22-10, Shenzhen Rayward LifeTechnology Co., Ltd, Shenzhen, China) under 1 L / min O ;

[0042] The rats were then placed on an electric heating pad to minimize the impact of the experimental operating table on the body temperature and other physiological factors of the experimental rats. The hair in the target area and surrounding areas of the stimulation was shaved clean and completely removed with a depilatory cream (Veet). The target area was determined according to the physiological anatomy of the rats, and the LIPUS probe (Chongqing Rong Hai Engineering Research Center of Ultrasound Medicine Co., Ltd, Chongqing, China) was placed in the target area (specifically the liver), and then ultrasonic stimulation or sham stimulation could be started. When LIPUS was working, the signal was transmitted between the LIPUS transducer and the target area through an ultrasonic coupling agent (Tianjin Chengxin Medical Auxiliary Materials Factory, Tianjin, China); the sham control included all steps of the LIPUS treatment, except that the LIPUS transducer was activated during the stimulation (the transducer power was turned off); the depth of the probe relative to the rat was always kept constant during the treatment (the low-intensity ultrasound transducer was kept at a fixed height above the laboratory workbench) to ensure that no pressure was applied to the organ through the transducer;

[0043] After LIPUS stimulation or sham stimulation was completed, the rats were observed to wake up from anesthesia and then placed back in the corresponding cage during the experiment.

[0044] 2. Effects of LIPUS irradiated liver tissue on blood glucose

[0045] During the experiment, a handheld blood glucose meter (Sinocare GA-3, Sinocare Inc., Changsha, China) was used to record the circulating blood glucose concentration and fasting blood glucose concentration of the rats. Figure 2 As shown in A and 2B, the weight of rats in each group was recorded during the experiment. Figure 2 As shown in C.

[0046] After the experiment, the rats were fasted for 12 hours, and the fasting blood glucose results were recorded. Figure 2 D, then a single oral administration of 2 g / kg glucose solution was performed by gavage with an oral syringe. After glucose administration, blood glucose was recorded by taking blood from the tail vein at 30 min, 60 min, 90 min, and 120 min. AUC-OGTT was calculated based on the AUC of glucose levels at baseline and 120 min after glucose overload. The results are shown in Figure 2 As shown in E.

[0047] The fasting serum insulin concentration was tested using a commercial ELISA kit, and the results are shown in 2F. The fasting glucose concentration was tested and HOMA was calculated according to the formula HOMA = fasting insulin (μU / mL) × fasting glucose (mmol / L) / 22.5. The results are shown in Figure 2 As shown in G.

[0048] Depend on Figure 2 A and 2B show that during the 30-day test period, the blood glucose of rats in the normal group (Normal group) was always within the normal range, and the blood glucose concentration of rats in the model group (Model group) gradually increased from a mean of 20.8mmol / L to a mean of 28.3mmol / L. In comparison, after daily LIPUS irradiation treatment, the peripheral blood glucose of diabetic rats decreased by about 3-5mmol / L after 30 days, and fasting blood glucose also decreased, which was comparable to the blood glucose reduction level of rats in the EMT group and LIPUS+MET group, and was statistically different from the model group (Model group).

[0049] Depend on Figure 2C shows that the weight of rats in the normal group (Normal group) increased by about 33.63g / rat, the weight of rats in the model group (Model group) decreased by about 28.32g / rat, the weight of rats in the LIPUS group decreased by about 16.84g / rat, the weight of rats in the MET group decreased by about 0.4g / rat, and the weight of rats in the MET+LIPUS group decreased by about 3.16g / rat. The weight of rats in the model group (Model group) was significantly lower than that in the normal group (Normal group). Unlike the situation in which the weight of rats in the model group (Model group) gradually decreased with the severity of the disease, the weight of rats in each treatment group showed a relative increase trend over time.

[0050] Depend on Figure 2 D and 2E show that oral administration of glucose did not cause significant changes in the blood glucose levels of rats in the normal group (Normal group), and the AUC at 120-min intervals did not change significantly. Compared with the rats in the normal group (Normal group), the fasting blood glucose levels of rats in the model group (Model group) were significantly increased (time 0 min), and the glucose tolerance to exogenous glucose was significantly impaired. In addition, compared with the model group (Model group), each treatment group significantly reduced blood glucose levels within 120 min. The 120-min integrated area under the blood glucose curve (AUC glucose ) was significantly higher than that of the normal group. Compared with the model group, the AUC glucose Significantly reduced.

[0051] Depend on Figure 2 F and 2G show that the serum insulin level of the rats in the model group decreased due to STZ administration. After 30 days of treatment, the serum insulin level of the rats in the model group further decreased, while the LIPUS treatment group prevented the deterioration of this situation, that is, prevented the decrease of serum insulin, and the effect of the MET group was particularly insignificant.

[0052] In summary, Figure 2 The results showed that LIPUS irradiation of liver tissue could prevent the decrease of serum insulin.

[0053] 3. LIPUS can reduce the incidence and severity of DC

[0054] After the rat T2DM model was successfully established, sodium pentobarbital, a mydriatic drug, was dripped into the rat's eyes. After the rat's pupil was fully dilated, the degree of lens opacity could be observed under a slit lamp. The grading standard of the Oxford University Ophthalmology Laboratory can be referred to: Stage 0 is a transparent lens; Stage I is mild opacity, with vacuolar changes in the peripheral cortex of the lens; Stage II is moderate opacity, with vacuoles extending toward the center and foggy opacity in the nucleus of the lens; Stage III is severe opacity, with vacuoles extending to the nucleus and worsening foggy opacity in the nucleus; Stage IV is nuclear opacity and develops into complete cataract. The opacity index was calculated according to the formula: opacity index = number of eyes in each stage × eye stage / total number of eyes to quantitatively evaluate the degree of lens opacity. The results are shown in Table 1. Figure 3 As shown in A;

[0055] Immediately after euthanasia, the rats in each group were completely excised and placed in sterile saline, with the optic nerve placed upward and fixed with forceps. Then an incision was made on the optic nerve entering the eye, and the sclera was pulled back to expose the lens. Subsequently, the ciliary body fragments attached to the equatorial plane of the lens were gently removed with forceps. The lens tissue of the left eye was fixed with eyeball fixative, and the lens tissue of the right eye was stored in a -80℃ refrigerator;

[0056] The incidence and severity of cataracts in each experimental group during the same period of time were recorded using a slit lamp microscope. Figure 3 As shown in B and 3C;

[0057] After treatment, the ocular appearance of the LIPUS group and the model group was as follows: Figure 3 As shown in D.

[0058] Depend on Figure 3It can be seen that under the slit lamp microscope, it can be found that the rats in the model group (Model group) began to have a small number of peripheral vesicles in the lens tissue in the second week after the successful modeling, which is a primary phenomenon of cataracts in the lens tissue, while the lens tissues of the rats in the LIPUS group, MET group and LIPUS+MET group remained transparent and normal. In the following 4th, 6th and 8th weeks, the lens tissues of the rats in the model group (Model group) gradually developed into vesicles and cortical opacities and diffuse central opacities. Until the 10th week, 95% of the rats in the model group (Model group) had cataracts, of which 95% of the lenses were in stage 4 and 5% of the lenses were in stage 3. Interestingly, the onset time of the LIPUS group and the MET group was the 4th week after the successful modeling, and the onset time of the LIPUS+MET group was the 6th week. In addition, the DC development rate of rats in the LIPUS group, MET group and LIPUS+MET group was slow. As of the 10th week, the lens tissue of rats in the normal group was always transparent throughout the study without turbidity. The incidence rate of rats in each treatment group was: 47% in the LIPUS group, 61.5% in the MET group, and 40% in the LIPUS+MET group; and the distribution of the lens in each stage of rats in the LIPUS group was as follows: Grand4: 20%, Grand3: 10%, Grand2: 60%, Grand1: 10%. The results show that LIPUS irradiation of liver tissue can reduce the incidence and severity of cataracts in diabetic rats.

[0059] To further analyze the effectiveness of LIPUS treatment, the present application performed hematoxylin-eosin (H&E) staining, immunoblotting, and real-time quantitative polymerase chain reaction (qRT-PCR) to perform pathological analysis of the anterior capsule and equatorial part of the lens tissue. Figure 4 As shown;

[0060] The specific steps of hematoxylin-eosin (H&E) staining are as follows: fresh liver, pancreas and lens tissues were fixed with 4% paraformaldehyde at room temperature, embedded in paraffin, and cut into slices with a thickness of 4 μm for histological examination. After deparaffinisation and rehydration, the prepared paraffin sections were stained with hematoxylin (H) solution for 8 min, then immersed in 1% acidic ethanol (70% volume concentration ethanol solution, the ethanol solution containing hydrochloric acid, the volume concentration of hydrochloric acid is 1%) for 5 times, and rinsed with distilled water; then, the sections were stained with eosin (E) solution for 3 min, then dehydrated with graded alcohol, washed with xylene, and the morphological changes were observed under an optical microscope (Olympus, Tokyo, Japan) after staining. The results are shown in Figure 2. Figure 4 As shown in A.

[0061] The specific steps of immunoblotting analysis were as follows: total protein was extracted from lens tissue using RAPI:PMSF (100:1) solution, and protein concentration was quantified using a BCA kit (Beyotime, Beyotime Biotechnology Co, Ltd, Shanghai, China); protein was mixed with 5×SDS sample buffer (4:1) (Beyotime, Beyotime Biotechnology Co, Ltd, Shanghai, China), and 40±5ug of protein sample was loaded on a 12% SDS-PAGE gel; blots were separated by electrophoresis and blotted on a PVDF membrane; then, the membrane was incubated with 5% bovine serum albumin (Solarbio, Beijing Solarbio Science & Technology Co., Ltd, Beijing, China) at room temperature for 1 h, and incubated with the primary antibody (specifically rabbit anti-e-cadherin (1:1000)) at 4°C overnight; the PVDF membrane (Millipore) was washed and incubated with the secondary antibody (specifically rabbit anti-α-SMA (1:1000)) at room temperature for 1 h (the antibody dilution concentration was as recommended in the manual); finally, chemiluminescent colorimetric solution (4A Biotech, Suzhou Sizhengbai Biotechnology Co., LTD, Suzhou, China) was used to visualize the images, and the intensity of each band was determined using Image J software (National Institutes of Health, USA), and the relative protein expression levels of GAPDH (AF7021, Affinity, Affinity Biosciences, OH, USA, 1:1000) or β-actin (AF7018, Affinity, Affinity Biosciences, OH, USA, 1:1000) were quantitatively compared. The results are shown in Figure 2. Figure 4 B and 4C;

[0062] The specific steps of real-time quantitative polymerase chain reaction (qRT-PCR) analysis are as follows: total cell RNA was extracted using an RNA extraction kit (Vazyme Biotech Co., Ltd, Nanjing, China), and RNA was reverse transcribed into cDNA using a reverse transcription kit (TaKaRa Biotechnology Co., Ltd, Oita Prefecture, Japan), PCR reaction solution was prepared, template and primers were added, and PCR amplification was performed using a real-time quantitative polymerase chain reaction (qRT-PCR) instrument (Agilent Technologies Co., Ltd, CA). The reaction conditions were: pre-denaturation at 95°C for 5 min, denaturation at 95°C for 5 s, annealing and extension at 60°C for 30 s, and repeated 40 cycles. GAPDH was used as the internal reference gene, and the relative expression of the gene was calculated using the 2-ΔΔCT formula. The primer sequences used (synthesized by a biological company) are shown in Table 1, the PCR reaction system is shown in Table 2, and the results are shown in Table 2. Figure 4 D and 4E.

[0063] Table 1 Primer sequences

[0064]

[0065] Table 2 PCR reaction system

[0066]

[0067] Depend on Figure 4 It can be seen that the lens fibers in the epidermis of the rats in the Normal group and the LIPUS group were arranged in an orderly and tight manner. In the rats in the Model group, some lens epithelial cells migrated to the epidermis, and the cell morphology changed from elliptical to spindle-shaped. In addition, the lens fibers in the cortex were edematous and vacuoles were formed, which was a manifestation of mesenchymal transformation ( Figure 4 A). The expression of E-cadherin protein in rats in the model group (Model group) decreased, and the expression of α-SMA protein increased, while the expression of E-cadherin protein in rats in the LIPUS group was upregulated, and the expression of α-SMA protein was downregulated ( Figure 4 B, 4C). The qRT-PCR test results also confirmed this ( Figure 4 D and 4E). The results indicate that LIPUS irradiation of liver tissue plays an important role in protecting the pathological changes of lens tissue under the long-term high-glucose environment of the body.

[0068] 4. Safety verification of LIPUS treatment

[0069] To verify the safety of LIPUS treatment, after obtaining serum samples, this application uses IDEXX Catalyst One biochemical analyzer to analyze the levels of albumin (ALB), alanine aminotransferase (ALT), aspartate aminotransferase (AST), glutamyl transferase (GGT), total bilirubin (TBIL) and other related indicators in the collected serum samples. The results are as follows Figure 5 shown.

[0070] Depend on Figure 5 It can be seen that compared with the rats in the model group, the various indicators of the rats in the LIPUS group were closer to the expression levels of the corresponding indicators in the normal group. This result shows that LIPUS irradiation plays a repairing role in liver tissue damage caused by STZ.

[0071] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the technology may modify or change the above embodiments without violating the spirit and scope of the present invention. Therefore, all equivalent modifications or changes made by a person of ordinary skill in the art without departing from the spirit and technical ideas disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. Use of a pulsed ultrasound device in the preparation of a medical device for treating and / or preventing diabetic cataracts in a subject.

2. The use according to claim 1, characterized in that The pulsed ultrasound instrument is used to output 180-220 mW / cm 2 The average sound intensity.

3. The use according to claim 2, characterized in that The average sound intensity is output in a pulse mode.

4. The use according to claim 1, characterized in that The pulsed ultrasound machine is used to irradiate the liver of a subject.

5. The use according to claim 4, characterized in that The irradiation frequency is 8-12 min / day for at least 30 consecutive days.

6. The use according to claim 4, characterized in that During the irradiation process, the duty cycle is 15%-25%, and the ultrasonic frequency is 280-300kHz.

7. The use of pulsed ultrasound apparatus in the preparation of medical devices for increasing the expression level of the epithelial marker protein E-cadherin in subjects.

8. The use of pulsed ultrasound in the preparation of medical devices for reducing the expression level of interstitial marker protein α-SMA in subjects.

9. Use of a pulsed ultrasound device in the preparation of a medical device for delaying the progression of diabetic cataracts in a subject.

10. Use of a pulsed ultrasound device in the preparation of a medical device for reducing the incidence of diabetic cataracts in a subject.

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