Device for in-situ real-time in-vivo detection of local fibrosis degree after neoadjuvant chemoradiotherapy of rectal cancer and storage medium

Abstract

The invention relates to the technical field of intra-operative evaluation of rectal cancer surgical operations, in particular to a device for in-situ real-time in-vivo detection of local fibrosis degree after neoadjuvant chemoradiotherapy of rectal cancer and a storage medium. The device is composed of a miniature laser, a base, a hinged type light path, a portable imaging scanning probe and a suspension device, the front end of the portable imaging scanning probe is 20 cm long, the portable imaging scanning probe can enter the rectum, multiphoton autofluorescence and second harmonic imaging are carried out on rectal tumors and intestinal walls, 142 features (including morphological features, texture features and intensity features) of collagen are extracted, and the size of the portable imaging scanning probe is larger than that of the suspension device. According to the method, the local fibrosis degree after neoadjuvant chemoradiotherapy of the rectal cancer is analyzed, whether anastomotic leakage, anastomotic stenosis and postoperative rectum functions occur or not after an operation is pre-judged before or during the operation, and meanwhile, a storage medium can conduct data calculation, analysis, statistics, classification and storage on fibrosis quantitative information.

Description

Technical Field

[0001] The present invention relates to the technical field of intraoperative evaluation of rectal cancer surgery, and in particular to a device and a storage medium for in situ real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy of rectal cancer. Background Art

[0002] Colorectal cancer is one of the most common malignant tumors in the world. In 2022, the number of new cases worldwide was >1.92 million, and the number of deaths was >900,000. The incidence rate ranked third in the world, and the mortality rate ranked second. In my country, the number of new cases in 2022 was approximately 517,100, and the number of deaths was approximately 240,000. The incidence rate ranked second, and the mortality rate ranked fourth. In recent years, the incidence rate has shown a clear upward trend. Compared with Western countries, the incidence rate of colorectal cancer in my country shows the characteristics of "three highs and one low": high incidence of rectal cancer, high proportion of low-position tumors, high incidence in young people but low early diagnosis rate. Therefore, many patients with rectal cancer are already in the locally advanced stage (Locally advanced rectal cancer, LARC) when they seek medical treatment, that is, the TNM stage is T 3-4 N0M0 or T 1-4 N 1- 2M0. Due to the anatomical characteristics of the rectum and tumor infiltration, these patients often experience local recurrence after direct surgery.

[0003] Preoperative neoadjuvant chemoradiotherapy (nCRT) can reduce the local recurrence rate, increase the anal preservation rate, improve the tumor resection rate and prolong the disease-free survival. Therefore, it is recommended as a treatment option for LARC by the National Comprehensive Cancer Network (NCCN) guidelines. However, increasing evidence shows that neoadjuvant chemoradiotherapy can increase the incidence of complications after rectal cancer surgery. Among them, the most serious complication is anastomotic leakage (AL), with a reported incidence ranging from 5% to 20%. Other complications include anastomotic stenosis. Therefore, routine preventive ileostomy is required, which significantly affects the patient's quality of life. Therefore, it is crucial for surgeons to identify patients at high risk of AL as early as possible.

[0004] Collagen fibers, as the main component of the extracellular matrix (ECM), are the main contributors to the function of the extracellular matrix. In addition, collagen structure also affects blood supply and the functions of different cells. Type I collagen, the main component of collagen fibers, is one of the important components to ensure the mechanical stability of the anastomosis. A potential factor for the occurrence of AL is the change in collagen structure. Therefore, by judging the degree of fibrosis, the possibility of AL can be better judged, thereby identifying high-risk patients.

[0005] Several studies have shown that the occurrence of AL is not only affected by the surgeon's skills, but also by the patient's clinical and pathological characteristics. Many risk factors have been reported to be associated with AL, such as gender, age, preoperative albumin, nCRT, etc. However, it is still difficult to accurately predict AL based on clinical and pathological characteristics.

[0006] Intraoperative frozen biopsy is the most commonly used and effective intraoperative detection method in clinical practice. However, it has the following limitations: (1) Time-consuming: Each intraoperative frozen biopsy takes about 30 minutes; (2) Irreversible in vitro: Intraoperative frozen biopsy requires cutting off tissue for biopsy, which is irreversible and harmful to the patient; (3) Poor collagen imaging effect: HE staining used in intraoperative frozen biopsy is not strong enough to distinguish between cell structures and collagen fibers.

[0007] Among the current tissue collagen quantification methods, the most commonly used is the analysis of collagen (hydroxyproline) content. The main operation process is: the tissue section is dried to constant weight at 100°C and then hydrolyzed in a 6 mol / L HCl solution at 110°C for 18 hours; after the acid hydrolysis solution is evaporated, the residue is washed three times with deionized water, and the residual acid must be completely evaporated between each washing step to remove the residual acid; after the sample is dissolved and treated with ethyl acetate citrate buffer (pH 6), the hydroxyproline value is analyzed by colorimetry, and the analysis result multiplied by a fixed coefficient of 7.46 is the collagen content, in ug / mg. The operation process of this method is relatively cumbersome and time-consuming, and the collagen content level in the tissue is obtained in a general and indirect manner, lacking a direct and global evaluation of the microstructure and characteristics of collagen.

[0008] Single-photon confocal laser endoscopy (CLE) is a commonly used in-situ real-time in vivo detection method in clinical practice. However, single-photon confocal endoscopic optical biopsy faces two major challenges in practical application: 1. Shallow imaging depth: It is impossible to obtain some deep lesion information such as fibrotic lesions after neoadjuvant treatment of locally advanced rectal cancer. 2. Dependence on exogenous fluorescent dyes: namely, sodium fluorescein, which may cause varying degrees of adverse reactions in the human body, including yellowing of urine and skin, and even induce allergic reactions, such as dyspnea, severe hypersensitivity reactions, and even fatal consequences.

[0009] In summary, there is currently a lack of an excellent method in clinical practice to perform in situ and real-time in vivo detection of the rectum during surgery to qualitatively and quantitatively analyze collagen and determine the degree of local fibrosis. Summary of the invention

[0010] The present invention provides a device and storage medium for in situ and real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer. The device consists of five parts: a miniaturized laser, a base, an articulated optical path, a portable imaging scanning probe, and a suspension. The storage medium can calculate, analyze, count, classify, and store quantitative fibrosis information.

[0011] The technical solution adopted by the present invention is: a device for in situ real-time in vivo detection of the degree of local fibrosis of rectal cancer after neoadjuvant chemoradiotherapy, characterized by: comprising a miniaturized laser, a base, an articulated optical path, a portable imaging scanning probe, and a suspender.

[0012] As a further improvement of the present invention, the miniaturized laser uses an ultrafast pulse laser with a central wavelength of 780nm, provides laser pulses with a repetition frequency of 80MHz, an average power of 0 to 140mW, a time length of each laser pulse of 100fs, and a pulse energy of 1.7nJ;

[0013] As a further improvement of the present invention, a miniaturized laser, a laser controller, and a computer host are fixed in the base;

[0014] As a further improvement of the present invention, the articulated optical path is mounted on the base, and the laser light of the miniaturized laser is sent into the portable imaging scanning probe through the articulated optical path;

[0015] As a further improvement of the present invention, the portable imaging scanning probe is used to excite the second harmonic generation (SHG) and autofluorescence (TPEF) signals generated by the tissue sample, and focus on the sample through an objective lens, and the generated signals are collected by a photomultiplier tube and a sensor;

[0016] As a further improvement of the present invention, the suspender is composed of a support column, an adjustment mechanism, a lever arm, and two hook mechanisms, and is used to bear the weight of the articulated optical path and the portable imaging scanning probe, so that the articulated optical path can extend freely.

[0017] As a further improvement of the present invention, the miniaturized laser has two output ports, one of which is a 1560nm laser wavelength output of a ring erbium-doped fiber laser, which is converted into a 100fs laser pulse output of 780nm through dual-frequency generation.

[0018] As a further improvement of the present invention, the laser controller is connected to the computer host via a USB port, and the laser controller is connected to the miniaturized laser via a D-sub37 cable.

[0019] As a further improvement of the present invention, a plurality of movable casters and a plurality of support adjustment blocks are connected to the bottom of the base.

[0020] As a further improvement of the present invention, the articulated optical path is divided into four sections, including a 25-centimeter short arm, two 50-centimeter long arms and a tubular arm connected to a portable imaging scanning probe, and the articulated arms are connected by a total of six bearings; nine first-plane tuning mirrors and two pairs of 90-degree rotating prisms are provided in the articulated optical path, and the laser is refracted by the nine first-plane tuning mirrors and the two pairs of 90-degree rotating prisms and sent into the portable imaging scanning probe.

[0021] As a further improvement of the present invention, the principle of the portable imaging scanning probe is as follows: the laser is first injected into the galvanometer through two second plane tuning mirrors. After entering the galvanometer, the laser is focused by a tubular lens, and then calibrated by a scanning lens to enter the rear hole of the objective lens. Finally, the laser is focused on the sample through the objective lens to excite the tissue sample to generate SHG and TPEF signals. The generated SHG and TPEF signals are collected by the same objective lens and scanning lens. The spectroscope reflects the emission signal to the photomultiplier tube. A filter is inserted between the spectroscope and the photomultiplier tube to reflect light in the range of 600-700nm to the sensor for reflection imaging. Two filters are inserted before the photomultiplier tube performs signal selection. The SHG is 390 / 10BP and the TPEF is 550 / 80BP. The SHG and TPEF signals can be collected respectively for easy analysis.

[0022] As a further improvement of the present invention, the support column is fixedly connected to the base, the top of the support column is rotatably connected to the lever arm through an adjustment mechanism, and the adjustment mechanism is connected to the lever arm through an adjustment pull rod, the two hook mechanisms are located at the bottom of the lever arm and are slidably connected thereto, and the support column is connected to a display screen through a bracket.

[0023] As a further improvement of the present invention, the adjustment mechanism includes a support frame and a support top plate fixedly connected to the support column, a rotating column rotatably connected between the support frame and the support top plate, and the rotating column is fixedly connected to the force arm, a first drive motor is fixedly installed on one side of the support frame, a first gear is provided at the output end of the first drive motor, and a second gear meshing with the first gear is provided on the outer fixed sleeve of the rotating column.

[0024] As a further improvement of the present invention, the hook mechanism includes a side frame slidably connected to the force arm, a second drive motor and a third drive motor are fixedly connected to one side of the side frame, a plurality of rollers are rotatably connected to the top end of the side frame and the output shaft of the second drive motor is connected to one of the rollers, a rotating roller is rotatably connected to the bottom end of the side frame and the output shaft of the third drive motor is connected to the rotating roller, a lifting rope is wrapped around the outside of the rotating roller, and a hook is provided at the bottom end of the lifting rope for connecting an articulated optical path and a portable imaging scanning probe.

[0025] A storage medium for in situ and real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer, including one of a solid-state hard drive and a flash memory card, is used to store quantitative fibrosis data and related information, so as to quantitatively detect the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer, predict whether anastomotic leakage, anastomotic stenosis and postoperative rectal function will occur before and during surgery, and provide assistance to surgeons in making decisions before and during surgery.

[0026] Beneficial effects of the present invention: The device and storage medium for in situ and real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer of the present invention can enter the rectum, detect the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer in situ and real-time in vivo (including collagen fiber morphological characteristics, texture characteristics, and intensity characteristics), and provide a storage medium to store fibrosis quantitative data and related information, thereby quantitatively detecting the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer, predicting whether anastomotic leakage, anastomotic stenosis and postoperative rectal function will occur before or during surgery, and providing assistance to surgeons in making decisions before or during surgery (such as whether intraoperative preventive ileostomy is needed). BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 It is a schematic diagram of the overall structure of a device and a storage medium for in situ real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer according to the present invention;

[0028] Figure 2 It is a partial structural schematic diagram of a device for in situ real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer and a storage medium of the present invention;

[0029] Figure 3 It is a schematic diagram of the internal structure of a portable imaging scanning probe of a device for in situ real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy of rectal cancer and a storage medium of the present invention;

[0030] Figure 4 This is a miniaturized laser front panel diagram of a device for in situ real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy of rectal cancer and a storage medium of the present invention;

[0031] Figure 5 This is a device for in situ real-time in vivo detection of local fibrosis after neoadjuvant chemoradiotherapy of rectal cancer and a miniaturized laser rear panel of the storage medium of the present invention.

[0032] Figure 6 This is a schematic diagram of the connection between a device for in situ real-time in vivo detection of the degree of local fibrosis of rectal cancer after neoadjuvant chemoradiotherapy and a miniaturized laser, a laser controller, and a computer host of the storage medium of the present invention;

[0033] Figure 7 It is a device for in-situ real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy of rectal cancer and an articulated optical path internal laser path diagram of a storage medium of the present invention;

[0034] Figure 8 It is a collagen characteristic value catalogue diagram of a device for in situ real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy of rectal cancer and a storage medium of the present invention;

[0035] Fig. 9 This is a multiphoton imaging effect diagram of a device for in situ real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer and a storage medium of the present invention;

[0036] Fig.10 The present invention discloses a device for in situ real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer and a collagen prediction flow chart of anastomotic leakage of the storage medium.

[0037] As shown in the figure: 1. Miniaturized laser; 2. Base; 3. Articulated optical path; 3.1. Short arm; 3.2. Long arm; 3.3. Tube arm; 4. Portable imaging scanning probe; 4.1. Objective lens; 4.2. Photomultiplier tube; 4.3. Sensor; 4.4. Second plane tuning mirror; 4.5. Galvanometer; 4.6. Tubular lens; 4.7. Scanning lens; 4.8. Spectroscope; 4.9. Filter; 5. Suspension; 5.1. Support column; 5.2. Adjustment mechanism; 5.2.1. Support frame; 5.2.2. Support top Plate; 5.2.3, rotating column; 5.2.4, first drive motor; 5.2.5, first gear; 5.2.6, second gear; 5.3, lever arm; 5.4, hook mechanism; 5.4.1, side frame; 5.4.2, second drive motor; 5.4.3, third drive motor; 5.4.4, roller; 5.4.5, lifting rope; 5.4.6, hook; 5.5, adjusting rod; 5.6, bracket; 5.7, display screen; 6, laser controller; 7, computer host; 8, movable casters; 9, support adjustment block. DETAILED DESCRIPTION

[0038] The directional terms such as up, down, left, right, front, back, front, back, top, bottom, etc. mentioned or may be mentioned in this specification are defined relative to their structure and are relative concepts. Therefore, they may change accordingly according to their different locations and different usage states; therefore, these or other directional terms should not be interpreted as restrictive terms.

[0039] The singular forms "a", "an", and "the" used in this specification are intended to include the plural forms, unless the context clearly indicates otherwise. It should also be understood that the term "and / or" used herein refers to and includes any or all possible combinations of one or more of the associated listed items.

[0040] In order to make the technical problems, technical solutions and beneficial effects to be solved by this application more clearly understood, the present application is further described in detail below in conjunction with the accompanying drawings and implementation methods. It should be understood that the implementation methods described herein are only used to explain this application and are not used to limit this application.

[0041] The present invention provides the following Figure 1-7 The device shown is for in situ and real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer, and is composed of a miniaturized laser 1, a base 2, an articulated optical path 3, a portable imaging scanning probe 4, and a suspender 5. The miniaturized laser 1 adopts an ultrafast pulse laser with a central wavelength of 780nm, provides laser pulses with a repetition frequency of 80MHz, an average power of 0 to 140mW, a time length of each laser pulse of 100fs, and a pulse energy of 1.7nJ; the miniaturized laser 1, a laser controller 6, and a computer host 7 are fixed in the base 2; the articulated optical path 3 is installed on the base 2, and the laser of the miniaturized laser 1 is sent to the portable imaging scanning probe 4 through the articulated optical path 3; the portable imaging scanning probe 4 is used to excite the SHG and TPEF signals generated by the tissue sample, and focus on the sample through the objective lens 4.1, and the generated signal is collected by the photomultiplier tube 4.2 and the sensor 4.3; the suspender 5 is composed of a support column 5.1, an adjustment mechanism 5.2, a force arm 5.3, and two hook mechanisms 5.4, and is used to bear the weight of the articulated optical path 3 and the portable imaging scanning probe 4, so that the articulated optical path 3 can be freely extended.

[0042] like Figure 4-6 As shown, the miniaturized laser 1 in the present invention has two output ports, one of which is the 1560nm laser wavelength output of the ring erbium-doped fiber laser, which is converted into a 100fs laser pulse output of 780nm through dual-frequency generation; the laser controller 6 is connected to the computer host 7 through a USB port, and the laser controller 6 is connected to the miniaturized laser 1 through a D-sub37 line.

[0043] like Figure 1 As shown, in the present invention, a plurality of movable casters 8 and a plurality of support adjustment blocks 9 are connected to the bottom of the base 2.

[0044] like Figure 1 and Figure 7 As shown, the articulated optical path 3 in the present invention is divided into four sections, including a 25 cm short arm 3.1, two 50 cm long arms 3.2 and a tube arm 3.3 connected to the portable imaging scanning probe 4, and the articulated arms are connected by a total of six bearings; nine first plane tuning mirrors and two pairs of 90-degree rotating prisms are arranged in the articulated optical path 3, and the laser is sent into the portable imaging scanning probe 4 through the refraction of the nine first plane tuning mirrors and the two pairs of 90-degree rotating prisms.

[0045] like Figure 3 As shown, the principle of the portable imaging scanning probe 4 in the present invention is as follows: the laser is first injected into the galvanometer 4.5 through two second plane tuning mirrors 4.4. After entering the galvanometer 4.5, the laser is focused by the tubular lens 4.6, and then calibrated by the scanning lens 4.7 to enter the rear hole of the objective lens 4.1. Finally, the laser is focused on the sample through the objective lens 4.1 to excite the tissue sample to generate SHG and TPEF signals. The generated SHG and TPEF signals are collected by the same objective lens 4.1 and the scanning lens 4.7. The beam splitter 4.8 reflects the emission signal to the photomultiplier tube 4.2. A filter 4.9 is inserted between the beam splitter 4.8 and the photomultiplier tube 4.2 to reflect the light in the range of 600-700nm to the sensor 4.3 for reflection imaging. Two filters are inserted before the photomultiplier tube 4.2 for signal selection. The SHG is 390 / 10BP and the TPEF is 550 / 80BP. The SHG and TEP signals can be collected respectively for easy analysis.

[0046] like Figure 1 and Figure 2 As shown, in the present invention, the support column 5.1 is fixedly connected to the base 2, the top of the support column 5.1 is rotatably connected to the lever arm 5.3 through an adjusting mechanism 5.2, and the adjusting mechanism 5.2 and the lever arm 5.3 are connected through an adjusting pull rod 5.5, two hook mechanisms 5.4 are located at the bottom of the lever arm 5.3 and are slidably connected thereto, and the support column 5.1 is connected to a display screen 5.7 through a bracket 5.6.

[0047] like Figure 1 and Figure 2As shown, the adjustment mechanism 5.2 in the present invention includes a support frame 5.2.1 and a support top plate 5.2.2 fixedly connected to the support column 5.1, a rotating column 5.2.3 rotatably connected between the support frame 5.2.1 and the support top plate 5.2.2, and the rotating column 5.2.3 is fixedly connected to the force arm 5.3, a first drive motor 5.2.4 is fixedly installed on one side of the support frame 5.2.1, a first gear 5.2.5 is provided at the output end of the first drive motor 5.2.4, and a second gear 5.2.6 meshing with the first gear 5.2.5 is provided on the outer fixed sleeve of the rotating column 5.2.3.

[0048] like Figure 1 and Figure 2 As shown, the hook mechanism 5.4 includes a side frame 5.4.1 which is slidably connected to the force arm 5.3, a second drive motor 5.4.2 and a third drive motor 5.4.3 are fixedly connected to one side of the side frame 5.4.1, a plurality of rollers 5.4.4 are rotatably connected to the top of the side frame 5.4.1, and the output shaft of the second drive motor 5.4.2 is connected to one of the rollers 5.4.4, a rotating roller is rotatably connected to the bottom end of the side frame 5.4.1, and the output shaft of the third drive motor 5.4.3 is connected to the rotating roller, a hanging rope 5.4.5 is wound around the outside of the rotating roller, and a hanging hook 5.4.6 for connecting the articulated optical path 3 and the portable imaging scanning probe 4 is provided at the bottom end of the hanging rope 5.4.5.

[0049] A storage medium for in situ and real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer, including one of a solid-state hard drive and a flash memory card, is used to store quantitative fibrosis data and related information, so as to quantitatively detect the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer, predict whether anastomotic leakage, anastomotic stenosis and postoperative rectal function will occur before and during surgery, and provide assistance to surgeons in making decisions before and during surgery.

[0050] Embodiment 1

[0051] This embodiment provides a device for in situ and real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer, as described below.

[0052] 1. Miniaturized Laser

[0053] The laser uses an ultrafast pulse laser with a central wavelength of 780nm. The laser can provide laser pulses with a repetition frequency of 80MHz and an average power of 0 to 140mW. The duration of each laser pulse is about 100fs and the pulse energy is about 1.7nJ. Due to the short pulse duration, after focusing to a sub-micro spot size, the peak laser intensity at the focus is close to 1×1012W / cm2, which far exceeds the minimum laser intensity required to excite spontaneous fluorescence and second harmonic emission of most biological tissues.

[0054] As attached Figure 4 As shown, the front panel of the laser is shown. The laser has two output ports. The left side is the 1560nm laser wavelength output of the erbium-doped ring fiber laser ( Figure 4 A), a 1560nm laser is transmitted through the barium borate crystal in the laser, and converted into a 780nm 100fs laser pulse after dual frequency generation ( Figure 4 B), A switch is used on the back of the laser to control the output wavelength of the laser, and the laser source is fixed to the base by three short steel clamps located at the bottom of the laser.

[0055] As attached Figure 5 As shown in the figure, the functions and effects of each control unit on the rear panel of the laser controller are shown. The laser controller is connected to the computer host through a standard USB port. The laser controller is connected to the computer host through a D-sub37 cable ( Figure 5 The middle port 11) is connected to the laser.

[0056] As attached Figure 6 As shown, the connection between the laser, the laser controller and the computer host is shown. After the three are connected through the above interface, the power of the laser can be controlled through the graphical user interface pre-installed on the computer to adjust the power of the laser.

[0057] (ii) Base

[0058] The base is fixed with a laser, a laser controller and a computer host; the bottom of the base is connected with a plurality of movable casters and a plurality of supporting adjustment blocks.

[0059] (III) Articulated optical path

[0060] The laser emits laser light and enters the portable imaging scanning probe through the articulated optical path. The entire articulated optical path structure is shown in the attached figure. Figure 7 As shown in FIG. 1 , the articulated arm consists of 4 parts, namely, a 25 cm short arm, two 50 cm long arms and a 50 mm long tube arm connected to the portable imaging scanning probe. Each articulated arm is connected by 6 bearings ( Figure 7 The laser beam is transmitted through 9 plane tuning mirrors (BE1-BE6) in sequence. Figure 7 TM1-TM9) and 2 pairs of 90 degree rotating prisms ( Figure 7 STM1) is sent to the portable imaging scanning probe.

[0061] (IV) Portable imaging scanning probe

[0062] The internal structure diagram of the portable imaging scanning probe is shown in the attached figure. Figure 3As shown, the principle is as follows: the laser is first injected into the galvanometer through two second plane tuning mirrors. After entering the galvanometer, the laser is focused by the tubular lens, and then calibrated by the scanning lens to enter the rear hole of the objective lens. Finally, the laser is focused on the sample through the objective lens to excite the tissue sample to generate SHG and TPEF signals. The generated SHG and TPEF signals are collected by the same objective lens and scanning lens. The beam splitter reflects the emission signal to the photomultiplier tube. A filter is inserted between the beam splitter and the photomultiplier tube to reflect light in the range of 600-700nm to the sensor for reflection imaging. Two filters are inserted before the photomultiplier tube performs signal selection. The SHG is 390 / 10BP and the TPEF is 550 / 80BP. The SHG and TEP signals can be collected respectively for easy analysis.

[0063] (V) Suspension

[0064] The suspender is composed of a support column, an adjustment mechanism, a lever arm, and two hook mechanisms. The specific structure is shown in the attached Figure 1 and attached Figure 2 As shown, it is used to bear the weight of the articulated optical path and the portable imaging scanning probe, so that the articulated optical path can be freely extended.

[0065] Among them, the support column is fixedly connected to the base, the top of the support column is rotatably connected to the force arm through an adjustment mechanism, and the adjustment mechanism is connected to the force arm through an adjustment pull rod, the two hook mechanisms are located at the bottom of the force arm and are slidably connected to it, and the support column is connected to a display screen through a bracket.

[0066] The adjustment mechanism includes a support frame and a support top plate fixedly connected to the support column, a rotating column rotatably connected between the support frame and the support top plate, and the rotating column is fixedly connected to the force arm, a first drive motor is fixedly installed on one side of the support frame, an output end of the first drive motor is provided with a first gear, and a second gear meshing with the first gear is provided on the outer fixed sleeve of the rotating column. The angle of the force arm can be adjusted by driving the first drive motor, so that it can be used at multiple angles, thereby improving the flexibility and adaptability of the equipment.

[0067] The hook mechanism includes a side frame slidably connected to the force arm, a second drive motor and a third drive motor are fixedly connected to one side of the side frame, a plurality of rollers are rotatably connected to the top end of the side frame and the output shaft of the second drive motor is connected to one of the rollers, a rotating roller is rotatably connected to the bottom end of the side frame and the output shaft of the third drive motor is connected to the rotating roller, a lifting rope is wound around the outside of the rotating roller, a lifting rope is provided at the bottom end of the lifting rope for connecting an articulated optical path and a portable imaging scanning probe, the rollers can be driven to rotate by the drive of the second drive motor, so that the positions of the two hook mechanisms can be adjusted, thereby changing the position of the portable imaging scanning probe, and the rotating rollers can be driven to rotate by the drive of the third drive motor, so as to retract and release the lifting rope, and then the height of the portable imaging scanning probe is adjusted by the lifting hook, which is suitable for use in different scenarios.

[0068] Embodiment 2

[0069] This embodiment utilizes a device for in situ real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer provided in the first embodiment to perform quantitative analysis of collagen and predict the prognosis of patients, as described below.

[0070] After multiphoton microscopy imaging, quantitative analysis of collagen and prediction of patient prognosis were performed through the following steps: (1) collagen second harmonic imaging; (2) binary image generation and collagen feature extraction; (3) selection of collagen features and construction of collagen scores; (4) classification of patients based on screening results combined with clinical characteristics.

[0071] (I) Collagen Second Harmonic Imaging

[0072] The second harmonic generation emitted by collagen was captured by the multiphoton microscopy imaging device described above.

[0073] (II) Binary image generation and collagen feature extraction

[0074] Taking advantage of the fact that the second harmonic in multiphoton imaging is very sensitive to collagen, the SHG signal image is segmented into collagen pixels and background pixels using the Gaussian mixture model algorithm, and the image is converted into a collagen binary image using the fiber network extraction method. On this basis, 142 collagen feature values ​​including collagen morphological features and textural features such as collagen area, length, width, curvature, cross-linking density and collagen direction index are extracted, including two types of morphological features and textural features (see the attached figure for specific collagen feature values). Figure 8 ).

[0075] The morphological characteristics include collagen area, length, width, number, straightness, cross-link density, cross-link space, and collagen orientation index.

[0076] The texture features include six eigenvalues ​​of mean, standard deviation, skewness, kurtosis, energy and entropy based on the histogram-based approach, 80 eigenvalues ​​of contrast, correlation, energy and homogeneity generated by five pixel distances (N=1, 2, 3, 4, 5) and four different directions (0°, 45°, 90°, 135°) based on the gray-level co-occurrence matrix (GLCM), and 48 eigenvalues ​​obtained by convolving the SHG signal image in four dimensions and six directions through Gabor wavelet transform filtering, and calculating the mean and standard deviation of each level respectively. There are a total of 134 eigenvalues ​​in the texture features.

[0077] (III) Selection of collagen features and construction of collagen scores

[0078] LASSO binary logistic regression was used to select significant predictors from all collagen features, and then the predictors were weighted by their respective LASSO regression coefficients. Finally, the collagen score for each patient was calculated by linear combination in the original and validation cohorts.

[0079] (IV) Classify patients based on screening results combined with clinical characteristics

[0080] According to the collagen score of each patient, the patients were divided into a high-risk group and a low-risk group for anastomotic leakage.

[0081] In summary, it can be seen from the above-mentioned Embodiment 1 and Embodiment 2 that the imaging principle of the device for in situ real-time in vivo detection of the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer of the present invention adopts multiphoton imaging. Compared with single-photon imaging, the device has a deeper imaging depth and a higher magnification, and has a higher detection accuracy; at the same time, the device realizes in situ real-time in vivo imaging during rectal cancer surgery. Compared with intraoperative freezing, the device does not require tissue resection and can observe tissue collagen structure, distribution, etc. in real time, which reduces time loss and damage to patients and increases detection flexibility; and the device realizes quantitative analysis of the degree of local fibrosis in patients with rectal cancer after nCRT through multiphoton imaging, which provides a basis for screening high-risk patients after nCRT, thereby improving patient prognosis and realizing individualized treatment.

[0082] Therefore, the present invention can analyze the collagen of the rectal cancer microenvironment after nCRT in situ, in real time, and in vivo, and the collagen characteristic score calculated by the extracted collagen characteristic parameters can be used to predict prognosis and guide clinical diagnosis and treatment.

[0083] The above implementation modes are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned implementation modes, a person skilled in the art should understand that the technical solutions described in the aforementioned implementation modes can still be modified, or some of the technical features can be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the implementation modes of the present invention.

Claims

1. A device for in situ real-time in vivo detection of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer, characterized by: The device is composed of a miniaturized laser (1), a base (2), an articulated optical path (3), a portable imaging scanning probe (4), and a suspender (5). The miniaturized laser (1) uses an ultrafast pulse laser with a central wavelength of 780 nm, provides laser pulses with a repetition frequency of 80 MHz, an average power of 0 to 140 mW, a duration of each laser pulse of 100 fs, and a pulse energy of 1.7 nJ; A miniaturized laser (1), a laser controller (6), and a computer host (7) are fixed inside the base (2); The articulated optical path (3) is installed on the base (2), and the laser light of the miniaturized laser (1) is sent to the portable imaging scanning probe (4) through the articulated optical path (3); The portable imaging scanning probe (4) is used to excite the second harmonic generation (SHG) and autofluorescence (TPEF) signals generated by the tissue sample and focus them on the sample through the objective lens (4.1). The generated signals are collected by the photomultiplier tube (4.2) and the sensor (4.3); The suspender (5) is composed of a support column (5.1), an adjustment mechanism (5.2), a force arm (5.3), and two hook mechanisms (5.4), and is used to bear the weight of the articulated optical path (3) and the portable imaging scanning probe (4), so that the articulated optical path (3) can be freely extended.

2. The device for in situ real-time in vivo detection of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer according to claim 1, characterized in that: The miniaturized laser (1) has two output ports, one of which is a 1560nm laser wavelength output of a ring erbium-doped fiber laser, which is converted into a 780nm 100fs laser pulse output through dual-frequency generation.

3. The device for in situ real-time in vivo detection of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer according to claim 1, characterized in that: The laser controller (6) is connected to the computer host (7) via a USB port, and the laser controller (6) is connected to the miniaturized laser (1) via a D-sub37 cable.

4. The device for in situ real-time in vivo detection of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer according to claim 1, characterized in that: The bottom of the base (2) is connected to a plurality of movable casters (8) and a plurality of support adjustment blocks (9).

5. The device for in situ real-time in vivo detection of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer according to claim 1, characterized in that: The articulated optical path (3) is divided into four sections, including a 25-centimeter short arm (3.1), two 50-centimeter long arms (3.2), and a tube arm (3.3) connected to a portable imaging scanning probe (4), and the articulated arms are connected via a total of six bearings; nine first-plane tuning mirrors and two pairs of 90-degree rotating prisms are arranged in the articulated optical path (3), and laser light is sent into the portable imaging scanning probe (4) through refraction of the nine first-plane tuning mirrors and the two pairs of 90-degree rotating prisms.

6. The device for in situ real-time in vivo detection of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer according to claim 1, characterized in that: The principle of the portable imaging scanning probe (4) is as follows: the laser is first injected into the galvanometer (4.5) through two second plane tuning mirrors (4.4). After entering the galvanometer (4.5), the laser is focused by the tubular lens (4.6), and then calibrated by the scanning lens (4.7) before entering the rear hole of the objective lens (4.1). Finally, the laser is focused on the sample through the objective lens (4.1), and the tissue sample is stimulated to generate SHG and TPEF signals. The generated SHG and TPEF signals are emitted by the same objective lens (4.1) and the scanning lens (4.7). .7) collection, the spectroscope (4.8) reflects the emission signal to the photomultiplier tube (4.2), a filter (4.9) is inserted between the spectroscope (4.8) and the photomultiplier tube (4.2), and the light in the range of 600-700nm is reflected to the sensor (4.3) for reflection imaging, and two filters are inserted before the photomultiplier tube (4.2) for signal selection, SHG is 390 / 10BP, and TPEF is 550 / 80BP, so that SHG and TPEF signals can be collected respectively for easy analysis.

7. The device for in situ real-time in vivo detection of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer according to claim 1, characterized in that: The support column (5.1) is fixedly connected to the base (2); the top of the support column (5.1) is rotatably connected to the lever arm (5.3) via an adjustment mechanism (5.2); the adjustment mechanism (5.2) and the lever arm (5.3) are connected via an adjustment pull rod (5.5); the two hook mechanisms (5.4) are located at the bottom of the lever arm (5.3) and are slidably connected thereto; the support column (5.1) is connected to a display screen (5.7) via a bracket (5.6).

8. The device for in situ real-time in vivo detection of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer according to claim 7, characterized in that: The adjustment mechanism (5.2) comprises a support frame (5.2.1) and a support top plate (5.2.2) fixedly connected to the support column (5.1); a rotating column (5.2.3) rotatably connected between the support frame (5.2.1) and the support top plate (5.2.2); and the rotating column (5.2.3) is fixedly connected to the force arm (5.3); a first drive motor (5.2.4) is fixedly mounted on one side of the support frame (5.2.1); a first gear (5.2.5) is provided at the output end of the first drive motor (5.2.4); and a second gear (5.2.6) meshing with the first gear (5.2.5) is provided on the outer fixed sleeve of the rotating column (5.2.3).

9. The device for in situ real-time in vivo detection of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer according to claim 7, characterized in that: The hook mechanism (5.4) comprises a side frame (5.4.1) slidably connected to the force arm (5.3); a second drive motor (5.4.2) and a third drive motor (5.4.3) are fixedly connected to one side of the side frame (5.4.1); a plurality of rollers (5.4.4) are rotatably connected to the top of the side frame (5.4.1), and the output shaft of the second drive motor (5.4.2) is connected to one of the rollers (5.4.4); a rotating roller is rotatably connected to the bottom of the side frame (5.4.1), and the output shaft of the third drive motor (5.4.3) is connected to the rotating roller; a hanging rope (5.4.5) is wound around the outside of the rotating roller; and a hanging hook (5.4.6) for connecting the articulated optical path (3) and the portable imaging scanning probe (4) is provided at the bottom of the hanging rope (5.4.5).

10. A storage medium for in situ real-time in vivo detection of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer, characterized by: It includes a solid-state drive and a flash memory card, which are used to store quantitative fibrosis data and related information, so as to quantitatively detect the degree of local fibrosis after neoadjuvant chemoradiotherapy for rectal cancer, predict whether anastomotic leakage, anastomotic stenosis and postoperative rectal function will occur before and during surgery, and provide assistance to surgeons in making decisions before and during surgery.

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