An experimental data anomaly automatic correction method and system about HSPA8

Abstract

This invention discloses an automatic correction method and system for experimental data anomalies related to HSPA8, belonging to the field of bioinformatics processing technology. The method includes: acquiring HSPA8 detection data, homology perturbation data, and Phlda3 association data to construct a standardized dataset; decoupling the perturbation data to construct degradation activity parameters; combining interaction deviation indices and degradation activity parameters to generate anomaly correction parameters; performing weighted fusion and boundary truncation based on the anomaly correction parameters; and outputting corrected data after double residual envelope discrimination. This method distinguishes between real metabolic changes and experimental physical errors at the mechanistic level, prevents extreme abnormal samples from causing abrupt numerical changes, ensures that the reconstructed results strictly conform to the objective physical laws of protein metabolism, and effectively improves the accuracy and reliability of quantitative analysis under complex perturbation environments.

Description

Technical Field

[0001] This invention relates to the field of bioinformatics processing technology, and in particular to an automatic correction method and system for experimental data anomalies related to HSPA8. Background Technology

[0002] In molecular biology mechanism studies, Western blotting is often used to assess the expression level of target proteins (such as HSPA8) and their spatial co-expression relationships with associated proteins (such as Phlda3). To investigate the metabolic patterns of specific proteins, researchers typically introduce MG132 (a proteasome inhibitor) and CHX (a protein synthesis inhibitor) to perform homology perturbation experiments to observe the stabilization accumulation and decay kinetics of proteins.

[0003] However, Western blotting experiments are cumbersome and highly susceptible to systematic physical errors such as fluctuations in lysis buffer titers, non-specific antibody binding contamination, and uneven membrane development, leading to frequent non-linear distortions in the extracted band grayscale data. Existing data processing techniques typically only perform static grayscale quantification and simple linear background subtraction, failing to differentiate at the biological mechanism level whether abnormal fluctuations in the data stem from genuine degradation kinetic changes or from experimental system failures or antibody contamination artifacts. Lacking a cross-validation mechanism that combines spatial interaction ratios with temporal degradation trajectories, current techniques are prone to producing erroneous quantification results when dealing with complex experimental data exhibiting background drift, potentially leading to serious misjudgments in downstream mechanism analysis. Summary of the Invention

[0004] In view of the aforementioned existing problems, the present invention is proposed.

[0005] Therefore, this invention provides an automatic correction method for experimental data anomalies related to HSPA8, which solves the technical problem that existing protein blot data processing techniques cannot distinguish between real metabolic changes and experimental physical errors at the biological mechanism level, resulting in nonlinear distortion and misjudgment of downstream mechanisms in the quantitative data of target proteins under complex perturbation environments.

[0006] To solve the above-mentioned technical problems, the present invention provides the following technical solution:

[0007] In a first aspect, the present invention provides an automatic correction method for experimental data anomalies related to HSPA8, which includes: acquiring HSPA8 protein blot detection data from the same biological sample source, homology perturbation response data based on MG132 and CHX, and Phlda3 association data at the 13891 Da position, and performing normalization processing to obtain a standardized dataset.

[0008] Decoupling analysis was performed on the homogeneous perturbation response data in the standardized dataset to obtain the stabilized release amount and the decay loss amount, and a degradation activity parameter reflecting the degradation potential of HSPA8 was constructed.

[0009] For the HSPA8 protein Western blot detection data and Phlda3 association data in the standardized dataset, a ratio calculation and deviation assessment are performed to obtain the interaction deviation index. The interaction deviation index is then coupled with the degradation activity parameter to generate anomaly correction parameters.

[0010] Based on the aforementioned anomaly correction parameters, the HSPA8 protein blot detection data are weighted and fused and reconstructed. The double residual envelope of MG132 and CHX is used to perform consistency discrimination on the fusion and reconstruction results, and the corrected HSPA8 experimental data are output.

[0011] As a preferred embodiment of the automatic correction method for experimental data anomalies regarding HSPA8 described in this invention, the normalization process to obtain a standardized dataset includes splitting protein lysates from the same biological sample into equal volumes to form a basic detection sample, an MG132-treated sample, and a CHX-treated sample, respectively.

[0012] Under the same Western blot detection conditions, Western blot imaging was performed on the basic detection sample, the MG132-treated sample, and the CHX-treated sample. The migration interval corresponding to the HSPA8 main band and a preset migration interval centered at 13891 Da were located in each lane, and the integrated grayscale values ​​of the corresponding bands were extracted. At least three background sampling windows far from the band region were selected in each lane to obtain the background grayscale values ​​and the total protein integral value of the corresponding lane. For the CHX-treated sample, Western blot imaging was performed at multiple preset time points, and the integrated grayscale values ​​of the corresponding lanes at each time point were extracted to form CHX time-series response data.

[0013] Based on the strip integral gray value, background gray value, and total protein integral value, HSPA8 protein blot detection data, MG132 perturbation response data, CHX time series response data, and Phlda3 association data were constructed by combining background gray median subtraction and total protein normalization. The above data were then subjected to uniform standardization processing to obtain standardized data, and a standardized dataset was constructed.

[0014] As a preferred embodiment of the automatic correction method for experimental data anomalies related to HSPA8 described in this invention, the steps of obtaining the stabilized release amount and the decay loss amount respectively include: calculating the relative change rate of the MG132 perturbation response data in the standardized dataset relative to the HSPA8 protein blot detection data, and performing nonlinear logarithmic dimensionality reduction on the relative change rate to extract the stabilized release amount; extracting the logarithmic decay amount of the CHX time-series response data in the standardized dataset at each preset time point relative to the HSPA8 protein blot detection data, allocating progressive weights according to the span ratio of each preset time point in the overall observation time window, and performing time-series accumulation on the weighted logarithmic decay amount to extract the decay loss amount.

[0015] As a preferred embodiment of the automatic correction method for experimental data anomalies regarding HSPA8 described in this invention, the construction of the degradation activity parameter includes: constructing a degradation amplitude correction factor using the stabilized release amount, and performing decoupling and reconstruction processing on the decay loss amount and the stabilized release amount by calculating the ratio of the decay loss amount to the degradation amplitude correction factor to generate the degradation activity parameter.

[0016] As a preferred embodiment of the automatic correction method for experimental data anomalies regarding HSPA8 described in this invention, the method for obtaining the interaction deviation index includes: calculating the relative expression ratio based on the Phlda3 association data corresponding to the basic detection in the standardized dataset and the HSPA8 protein Western blot detection data corresponding to the basic detection.

[0017] The relative expression ratio is compared with a preset reference ratio center to obtain the deviation; after normalizing the deviation, it is substituted into the hyperbolic tangent function to perform bidirectional soft saturation nonlinear mapping to obtain the interaction deviation index.

[0018] As a preferred embodiment of the automatic correction method for experimental data anomalies regarding HSPA8 described in this invention, the generation of anomaly correction parameters includes: assigning preset weight coefficients to the interaction deviation index and the degradation activity parameter respectively and performing a linear combination, substituting the result of the linear combination as a negative exponential term into the logistic regression mapping model to perform probability transformation processing, and using the processing result as the anomaly correction parameters.

[0019] As a preferred embodiment of the automatic correction method for experimental data anomalies related to HSPA8 described in this invention, the weighting and fusion reconstruction of HSPA8 protein blot detection data includes: using degradation activity parameters to recalibrate the potential energy of MG132 perturbation response data in the standardized dataset to obtain a correction reference value.

[0020] Using the anomaly correction parameter as the fusion weight, linear interpolation fusion was performed between the HSPA8 protein blot detection data and the corrected reference value to obtain the fusion reconstruction result;

[0021] The consistency judgment of the fusion reconstruction results includes calculating the stabilization direction residual of the fusion reconstruction results relative to the MG132 disturbance response data, and the decay direction residual of the fusion reconstruction results relative to the corresponding decay trend of the CHX time series response data.

[0022] Preset penalty weights are assigned to the stabilization direction residual and the decay direction residual respectively and combined to obtain the double residual envelope score; the double residual envelope score is compared with a preset consistency threshold, and when the double residual envelope score is greater than or equal to the consistency threshold, the fusion reconstruction result is output as the corrected HSPA8 experimental data.

[0023] The corrected HSPA8 experimental data includes fusion reconstruction results, anomaly correction parameters, and double residual envelope scores.

[0024] Secondly, the present invention provides an automatic correction system for experimental data anomalies related to HSPA8, comprising: a data module, which acquires HSPA8 protein blot detection data from the same biological sample source, homology perturbation response data based on MG132 and CHX, and Phlda3 association data at the 13891 Da position, and performs normalization processing to obtain a standardized dataset.

[0025] The analysis module performs decoupling analysis on the homogeneous perturbation response data in the standardized dataset to obtain the stabilized release amount and the decay loss amount, and constructs degradation activity parameters that reflect the degradation potential of HSPA8.

[0026] The anomaly correction module performs ratio calculation and deviation assessment on the HSPA8 protein Western blot detection data and Phlda3 association data in the standardized dataset to obtain the interaction deviation index, and performs coupling processing on the interaction deviation index in combination with the degradation activity parameter to generate anomaly correction parameters.

[0027] The execution module performs weight allocation and fusion reconstruction on the HSPA8 protein blot detection data according to the anomaly correction parameters, and uses the double residual envelope of MG132 and CHX to perform consistency judgment on the fusion reconstruction results, and outputs the corrected HSPA8 experimental data.

[0028] Thirdly, the present invention provides a computer device including a memory and a processor, wherein the memory stores a computer program, wherein: when the computer program is executed by the processor, it implements any step of the automatic correction method for experimental data anomalies regarding HSPA8 as described in the first aspect of the present invention.

[0029] Fourthly, the present invention provides a computer-readable storage medium having a computer program stored thereon, wherein: when the computer program is executed by a processor, it implements any step of the automatic correction method for experimental data anomalies regarding HSPA8 as described in the first aspect of the present invention.

[0030] The beneficial effects of this invention are as follows: By standardizing the HSPA8 protein blot detection data, MG132 perturbation response data, and CHX time-series response data of homologous biological samples, and combining this with Phlda3 correlation data to construct a multidimensional data foundation, systematic errors caused by differences in experimental batches, antibody titer fluctuations, and development conditions can be effectively eliminated. Based on this, by decoupling the stabilization release amount and decay loss amount, degradation activity parameters reflecting the true degradation potential of the protein are constructed, achieving quantitative characterization of the dynamic metabolic characteristics of the protein. Simultaneously, a nonlinear deviation evaluation mechanism based on hyperbolic tangent mapping is introduced to constrain the relative expression relationship between HSPA8 and Phlda3, effectively suppressing the amplified influence of abnormally high noise signals on the formulation results. Furthermore, by combining degradation activity parameters and interaction deviation indices, anomaly correction parameters are generated through a logistic regression model, achieving fusion decision-making of spatial and temporal dimensions. Finally, through weight allocation and fusion reconstruction combined with a double residual envelope discrimination mechanism, biologically consistent correction results are output. This improves the reliability and interpretability of protein experimental data and reduces the impact of human error and non-specific interference on the results. Attached Figure Description

[0031] To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the following description of the embodiments will be briefly introduced. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0032] Figure 1 This is a flowchart of an automatic correction method for experimental data anomalies related to HSPA8.

[0033] Figure 2 This is a schematic diagram of the degradation activity parameter generation process for an automatic correction method for experimental data anomalies related to HSPA8.

[0034] Figure 3 This is a diagram of a computer setup for an automatic correction method for experimental data anomalies related to HSPA8. Detailed Implementation

[0035] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

[0036] Many specific details are set forth in the following description in order to provide a full understanding of the invention. However, the invention may also be practiced in other ways different from those described herein, and those skilled in the art can make similar extensions without departing from the spirit of the invention. Therefore, the invention is not limited to the specific embodiments disclosed below.

[0037] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The phrase "in one embodiment" appearing in different places in this specification does not necessarily refer to the same embodiment, nor is it a single or selective embodiment that is mutually exclusive with other embodiments.

[0038] Reference Figures 1-3 As one embodiment of the present invention, this embodiment provides an automatic correction method for experimental data anomalies related to HSPA8, comprising the following steps:

[0039] S1: Obtain HSPA8 protein blot detection data, homology perturbation response data based on MG132 and CHX, and Phlda3 association data at the 13891 Da position from the same biological sample, and perform normalization processing to obtain a standardized dataset.

[0040] In this embodiment, "same biological sample source" means that the protein lysis buffers used to form the basic detection sample, MG132-treated sample, and CHX-treated sample all originate from the same batch, the same treatment background, and the same sample collection time of the target biological sample, rather than from spliced ​​samples from different culture batches, different animal individuals, or different time points. This ensures that the subsequently obtained HSPA8 protein blot detection data, MG132 perturbation response data, CHX time-series response data, and Phlda3 association data are biologically homologous, avoiding the misinterpretation of differences in the basic state between different samples as abnormal signals.

[0041] In practice, the target sample is first selected and a total protein lysis buffer is prepared. This embodiment consistently uses AR42J cells as the example object. The same batch of cells is collected under the same culture conditions, and total protein is extracted using RIPA lysis buffer. After centrifugation to remove cell debris, the supernatant is obtained as the total protein lysis buffer. The AR42J cells, RIPA lysis buffer, Western blotting, and subsequent CHX and MG132 treatment conditions are disclosed in the document.

[0042] After obtaining the total protein lysate, the lysate was divided into three groups of equal volume. Based on the protein concentration determination results, the total protein content participating in subsequent electrophoresis loading was kept consistent across the three groups, rather than simply being distributed by volume. The three groups were the basic detection group, the MG132 treatment group, and the CHX treatment group.

[0043] The basic detection group refers to the sample group used to collect the basic HSPA8 band and Phlda3 band without additional proteasome inhibition or protein synthesis inhibition treatment; the MG132 treatment group refers to the sample group used to characterize the HSPA8 stabilization response after the addition of MG132; the CHX treatment group refers to the sample group used to form the HSPA8 time-decay sequence after the addition of CHX at multiple preset time points.

[0044] The basic detection group provides the original state, the MG132 treatment group provides the response state under the background of proteasome inhibition, and the CHX treatment group provides the dynamic change state after protein synthesis is blocked. The three together constitute the input basis for subsequent automatic correction of abnormalities.

[0045] For the MG132 treatment group, MG132 was added to the sample to a final concentration of 10 μM and the treatment was continued for 6 hours. The validity of the MG132 treatment group was determined by whether the sample group met two conditions: first, the source of the samples used before and after treatment was completely consistent; second, except for MG132, the other culture, temperature, and incubation time conditions remained unchanged during the treatment.

[0046] For the CHX treatment group, CHX was added to the samples at a final concentration of 100 μg / mL, and samples were collected at 0, 2, 4, and 6 hours. CHX time-series response data were generated by repeatedly performing Western blot analysis on the same group of samples at fixed time points after CHX addition, and extracting the integrated gray values ​​of the corresponding bands from each time point.

[0047] After sample processing, the samples from the basic detection group, MG132 treatment group, and CHX at each time point were subjected to SDS-PAGE electrophoresis, membrane transfer, blocking, antibody incubation, and ECL chemiluminescence imaging, and Western blot images were acquired under the same imaging parameters.

[0048] Under identical Western blot detection conditions, the following criteria must be met: consistent gel concentration; consistent transfer procedure; consistent primary and secondary antibody sources and dilution ratios; consistent ECL developing reagent; consistent imaging equipment; and consistent exposure rules. Only when these conditions are consistent can the subsequently extracted band integrated gray values ​​be horizontally comparable.

[0049] After image acquisition, band window localization is performed for each lane. HSPA8 protein blot detection data refers to the raw grayscale data obtained by integrating the migration interval corresponding to the main HSPA8 band within the lane corresponding to the basic detection group; Phlda3 associated data refers to the raw grayscale data obtained by integrating the preset migration interval centered at 13891 Da within the lane corresponding to the basic detection group. Here, "13891 Da position" should not be understood as an absolute pixel, but rather as a "preset band interval centered at the migration position corresponding to 13891 Da"; the theoretical molecular weight corresponds to the migration range, not a fixed single point on the image; if the band actually falls within this preset migration interval, it can be determined that the acquisition condition of "Phlda3 associated data at the 13891 Da position" is met.

[0050] After locating the HSPA8 and Phlda3 band windows, it is necessary to select a background sampling window and a total protein baseline region in each lane. A background sampling window is a blank image area located within the same lane, far from the target band region, and not overlapping with other visible bands. "Far from the band region" is defined as the background window boundary not overlapping with the HSPA8, Phlda3, or other clearly visible band windows. If a background window shows obvious bright spots, trailing, or other band afterimages, it is considered invalid and needs to be reselected. At least three background sampling windows mean that at least three separate background windows are set in each lane, and their grayscale values ​​are extracted separately.

[0051] Within the same lane, the total protein integral value also needs to be extracted. The total protein integral value refers to the overall integrated grayscale of the total protein stained area within the same lane, used to reflect the total protein loading in that lane. After transfer, the entire lane is stained with total protein, and the overall integrated grayscale value of that lane is calculated by image analysis software. The reason for using the total protein integral value instead of a single internal control protein such as β-actin or GAPDH is that under conditions including CHX and MG132 perturbation, a single internal control protein may fluctuate, while the total protein value more stably reflects the sample loading amount.

[0052] In this context, HSPA8 represents the target protein to be analyzed for automatic anomaly correction; MG132 represents the proteasome inhibitor treatment conditions used to generate a stabilizing perturbation response; CHX represents the protein synthesis inhibitor treatment conditions used to generate a time-decrease response; 13891Da represents the position corresponding to the theoretical relative molecular weight of Phlda3; Phlda3 represents the associated protein that interacts with HSPA8; AR42J represents the target cell model used in the examples; RIPA represents the lysis buffer used to extract the total protein lysis buffer; SDS-PAGE represents the protein electrophoresis separation step; and ECL represents the chemiluminescent imaging method.

[0053] After completing band integration, background sampling, and total protein integration, standardized processing was performed on the basic detection group, MG132 treatment group, CHX samples at each time point, and Phlda3 band data. The HSPA8 standardized value in the basic detection group can be obtained using the following formula:

[0054]

[0055] in, This represents the HSPA8 normalized gray value of the basic test group; This represents the original integrated grayscale of the HSPA8 stripe in the basic detection group; med represents the median value of the background window. This represents the grayscale value of the background sampling window for the basic detection group; This represents the total protein score of the basic testing group; This represents a tiny constant to prevent the denominator from being zero.

[0056] The HSPA8 normalized value in the MG132 processing group can be obtained according to the following formula:

[0057]

[0058] in, This represents the HSPA8 normalized grayscale value of the MG132 processing group; This indicates the original integrated grayscale of the HSPA8 stripe corresponding to the MG132 processing group; This represents the grayscale value of the background sampling window for the MG132 processing group; This represents the total protein score of the MG132 treatment group.

[0059] The standardized HSPA8 values ​​of CHX samples at each time point can be obtained using the following formula:

[0060]

[0061] in, Indicates a point in time HSPA8 normalized grayscale values; This indicates the CHX processing group at time point. The original integral grayscale of the HSPA8 stripe below; Indicates the time sampling point of the CHX processing group; This indicates the CHX processing group at time point. The grayscale value of the background sampling window below; This indicates the CHX processing group at time point. The total protein score.

[0062] The standardized Phlda3 value in the basic test group can be obtained according to the following formula:

[0063]

[0064] in, This represents the Phlda3 normalized gray value of the basic detection group; This represents the original integrated gray level of the Phlda3 strip within the migration interval corresponding to 13891Da in the basic detection group.

[0065] After obtaining the standardized values ​​mentioned above, the HSPA8 standardized values ​​of the basic detection group, the HSPA8 standardized values ​​of the MG132 treatment group, the HSPA8 standardized values ​​of CHX at various time points, and the Phlda3 standardized values ​​of the basic detection group were combined to form a standardized dataset. The "standardized dataset" refers to a data set obtained after processing with a unified source, unified perturbation rules, unified imaging conditions, unified background subtraction method, and unified total protein normalization method. This standardized dataset includes the HSPA8 standardized values ​​of the basic state, the HSPA8 standardized values ​​of the MG132 treatment state, the HSPA8 standardized values ​​of CHX at multiple time points, and the Phlda3 standardized values ​​of the basic state.

[0066] The data construction method using samples from the same biological source ensures that basic detection, MG132 treatment, and CHX treatment are all performed under the same batch of lysis buffer conditions, avoiding baseline differences caused by different culture batches and sampling times. Traditional Western blot analysis often directly compares results from different experiments, which can easily misinterpret inherent differences between samples as expression abnormalities. Here, homology splitting is used to ensure that all subsequent data are consistent in biological context, reducing error propagation from the source.

[0067] At the image processing level, the integrated grayscale values ​​of the bands are obtained under the same electrophoresis, transfer, and development conditions. Simultaneously, a background sampling window and the total protein integral value are introduced as dual calibration benchmarks. Compared to methods relying solely on a single internal control protein, background median subtraction eliminates local development noise, and total protein normalization corrects for variations in sample loading. The combination of these two methods makes the grayscale values ​​more comparable and stable.

[0068] Further, by acquiring data at multiple time points using CHX, time-series response data were generated, transforming static protein bands into information sequences with a time dimension. This processing allows subsequent analysis to move beyond the limitations of expression intensity at a single moment and reflect the changes in protein over time, thus providing a data foundation for subsequent metabolic behavior analysis.

[0069] S2: Decouple the homologous perturbation response data in the standardized dataset to obtain the stabilization release amount and decay loss amount, and construct degradation activity parameters that reflect the degradation potential of HSPA8.

[0070] like Figure 2 As shown, basic detection data from the standardized dataset is used. MG132 disturbance response data And introduce the standard deviation of background noise. Perform a valid response determination; the This is the standard deviation of the background noise calculated from the grayscale values ​​of the background sampling window in step S1. The specific operation is as follows: the system calculates... The difference, and determine whether the difference is greater than If determined to be greater than If the MG132-induced stabilization response is confirmed to be valid, subsequent logarithmic compression calculations are triggered; if the result is determined to be less than or equal to... If the response is invalid, the amount of stable release will be determined. The value is directly assigned to zero. Assuming the response is valid, the system, in order to reduce the impact of extreme release samples on the model, executes the following formula for stabilizing the release amount extraction:

[0071]

[0072] in, This represents the stabilized release parameter extracted from the MG132 perturbation channel after decoupling.

[0073] CHX time series response data in the dataset Before performing time-series accumulation, the system first detects the internal reference protein at the end of the time points for analysis. The retention ratio is used to determine cytotoxicity cutoff. Specifically, the system retrieves the grayscale value of the internal reference protein at the end sampling point and calculates its proportion relative to the baseline state. If this proportion is below 0.75, it is determined that cell death has triggered a disruption of order. The 0.75 threshold is an empirical threshold obtained through frame-by-frame statistical analysis. After confirming that the temporal logic has not been disrupted, the system constructs an increasing non-uniform temporal weight based on the position of each time point within the overall observation window to highlight the contribution of the later-stage decay to the degradation rate. The following temporal weighted loss accumulation formula is then executed:

[0074]

[0075] in, This represents the decay loss parameter extracted from the CHX tracking channel, used to characterize the level of loss in overall sample stability without the replenishment of new protein; Indicates the total number of CHX sampling points; Indicates the index of the current time point; This represents the scale value of the last valid time point involved in the calculation, and its relation to... The ratio constitutes a non-linear weight that increases over time. The calculated... It fully reflects the natural decay trajectory of HSPA8 without synthetic intervention.

[0076] After acquiring the dynamic indicators of the two independent dimensions mentioned above, the system enters the decoupling and reconstruction stage of degradation potential energy. This involves reconstructing the quotient between "current loss" and "suppressible release," thus transforming the system from a phenomenological description to a quantifiable mechanism. Specifically, the system utilizes the obtained... Construct a degradation magnitude correction factor and calculate it. The ratio of this correction factor is used to construct the degradation activity parameter using the following formula:

[0077]

[0078] in, The degradation activity parameter, which represents the final generated degradation potential energy of HSPA8, has the physical significance of characterizing the true background degradation kinetic energy after removing the proteasome recovery factor. This appears and represents the attenuation loss calculated above; This represents the stabilized release amount calculated above. This operation achieves deep decoupling at the kinetic level: as the decaying loss... When the levels are greatly elevated but primarily attributed to the proteasome-sensitive pathway, synchronous increases occur. This will effectively smooth out the denominator. The value is used to accurately filter out abnormal epigenetic degradation caused by highly active ubiquitination.

[0079] We decoupled the MG132 and CHX perturbation data separately, separating the various influencing factors that were originally mixed in with a single grayscale change. The MG132 channel reflects the release changes of proteins under inhibited degradation, while the CHX channel reflects the decay process of proteins under inhibited synthesis. By processing these separate channels, the data features corresponding to different biological mechanisms can be extracted independently.

[0080] In the process of stabilizing release extraction, logarithmic compression is introduced to smooth out large grayscale changes and avoid abnormal amplification caused by oversaturation of development or sample loading deviation. Compared with directly using ratios or differences, compression significantly reduces the impact of extreme values ​​on the overall results, making the parameters more stable.

[0081] In calculating the decay loss, weights are allocated proportionally over time spans and accumulated over time, allowing later time points to have a greater impact on the overall decay trend, thus more accurately reflecting the dynamic characteristics of the protein degradation process. Finally, a degradation activity parameter is constructed using the ratio of release to loss, achieving a quantitative description of the driving force behind protein degradation and avoiding biases caused by a single indicator.

[0082] S3: Perform ratio calculation and deviation assessment on the HSPA8 protein blot detection data and Phlda3 association data in the standardized dataset to obtain the interaction deviation index, and perform coupling processing on the interaction deviation index in combination with the degradation activity parameter to generate anomaly correction parameters.

[0083] Upon receiving the degradation activity parameters, a quantitative verification mechanism for the co-expression relationship between HSPA8 and Phlda3 is immediately initiated. In the ratio calculation and deviation assessment operations, a nonlinear deviation quantification model based on hyperbolic tangent mapping is chosen. This is because if a conventional linear ratio is used for subtraction, the ratio difference will be infinitely amplified when high-intensity false positive spots appear in the developed image, leading to overcompensation or even collapse of the subsequent reconstruction algorithm. The hyperbolic tangent function, however, possesses a natural soft-saturation boundary constraint capability, enabling it to smoothly and symmetrically map any physical deviation to a fixed threshold range.

[0084] Specifically, the system extracts basic state data, calculates the real-time relative expression ratio of Phlda3 to HSPA8, compares the difference with the preset reference ratio center, performs normalization and compression, and executes interaction deviation evaluation calculation.

[0085]

[0086] in, The generated interaction deviation index is used to quantitatively characterize the physical degree to which the binding state of HSPA8 and Phlda3 in the current sample deviates from the normal biological axis, and its value range is strictly limited to -1 to 1. Represents the hyperbolic tangent mapping function; This indicates a preset reference ratio center, which is the average ratio of HSPA8 to Phlda3 obtained based on pre-experimental statistics. This represents a preset reference discrete scale, which is the standard deviation of the ratio obtained statistically from pre-experimental data. After the calculation, the system transforms the static image grayscale ratio into a spatial constraint index with a clear biological determination direction. .

[0087] After obtaining the interaction deviation index, the system fuses it with the degradation activity parameter. A heterogeneous parameter coupling decision model based on logistic regression is chosen for this coupling process because the interaction deviation index (characterizing spatial anomalies) and the degradation activity parameter (characterizing temporal kinetic anomalies) belong to completely different physical dimensions, and direct addition lacks mathematical basis. Introducing the exponential structure of logistic regression can compress these two heterogeneous multidimensional features into a continuous probabilistic weight between 0 and 1, perfectly matching the system's decision requirement for the next step of adaptive reconstruction.

[0088] Specifically, the system assigns preset weight coefficients to the interaction deviation index and degradation activity parameter, and performs a linear combination. Then, it substitutes these coefficients as negative exponential terms into the mapping function to perform probability transformation processing, generating anomaly correction parameters. The calculation is as follows:

[0089]

[0090] in, This represents the final generated anomaly correction parameters; Represents an exponential function composed of natural constants; This represents the preset interaction deviation weighting coefficient; The interaction deviation index appears and represents the value generated in the previous calculation step; This represents a preset degradation potential energy weighting coefficient; the weighting coefficient and All of these are deterministic constants obtained by fitting pre-experimental data with maximum likelihood estimation.

[0091] In the spatial dimension, by calculating the relative expression ratio of Phlda3 to HSPA8 and comparing it with a reference matching center, an index reflecting the degree of deviation from the interaction relationship is obtained. Traditional methods usually directly use the ratio to judge expression changes, which is easily affected by local noise; here, by introducing a reference matching center, the judgment is based on statistical regularities rather than single-point observations, thus improving stability.

[0092] In the deviation assessment process, a hyperbolic tangent mapping is introduced to compress the deviation, limiting the deviation value to a finite range and preventing high-intensity abnormal signals from excessively affecting the results. Compared with the linear interpolation method, this nonlinear mapping can effectively suppress extreme disturbances caused by factors such as developing black spots and background fluctuations.

[0093] In the fusion decision-making stage, interaction deviation indicators and degradation activity parameters are jointly processed, and spatial relationships and temporal dynamics information are unified into a single evaluation framework through logistic regression. Data from different sources are converted into continuous probability forms, enabling subsequent decisions to no longer rely on a single threshold judgment, but to provide stable results by integrating multidimensional information.

[0094] S4: Based on the anomaly correction parameters, the HSPA8 protein blot detection data are weighted and fused and reconstructed. The double residual envelope of MG132 and CHX is used to perform consistency discrimination on the fusion and reconstruction results, and the corrected HSPA8 experimental data are output.

[0095] Upon receiving the anomaly correction parameters, the system immediately initiates the numerical reconstruction and closed-loop verification procedure for the original HSPA8 detection data. In the specific operations of weight allocation and fusion reconstruction, a progressive weighted fusion model based on degradation potential energy recalibration is selected. This model is chosen because directly replacing the original detection values ​​with theoretically calculated values ​​would not only disrupt the original data distribution but also easily lead to abrupt numerical drops when encountering extreme anomalies, significantly reducing the algorithm's fault tolerance and robustness. In contrast, using a continuous weighted fusion model based on correction parameters can progressively correct for anomalies decoupled from biological mechanisms while fully preserving the original physical background of the detection data.

[0096] The specific operation involves the system extracting MG132 perturbation response data from the standardized dataset and recalibrating its background potential using degradation activity parameters to obtain a corrected reference value characterizing the upper limit of the ideal state. Subsequently, the system uses anomaly correction parameters as fusion weights to perform linear interpolation fusion between the HSPA8 protein blot detection data and the corrected reference value, and performs a fusion reconstruction operation.

[0097]

[0098]

[0099] in, This represents the corrected reference value obtained after recalibration based on the degradation potential energy, used to provide a theoretical upper limit anchor point for correction. This represents the MG132 disturbance response data; This represents the fusion and reconstruction result generated after weight allocation and linear interpolation; The HSPA8 protein blot detection data output from the preceding step S1 is generated and retrieved. Through this calculation, when... When the value is extremely small, the fusion and reconstruction results Seamless degradation is preserved as the original background; when When the value approaches 1, the result of fusion and reconstruction is... Automatic approximation of corrected reference values ​​strongly constrained by biological mechanisms .

[0100] After completing the integration and reconstruction, Subsequently, the system does not directly output this as the final result, but must perform a dual residual envelope consistency judgment operation on it. For this consistency judgment operation, this embodiment adopts a bidirectional envelope verification model based on the homologous perturbation boundary. The purpose of using this model is to introduce endogenous boundary constraints of the experimental system to prevent data distortion caused by over-correction of the algorithm. Specifically, since the MG132 perturbation response data reflects the upper limit of protein accumulation when the descent solution of the experimental system is blocked, and the CHX time-series response data reflects the background decay law in the absence of synthesis, if the fusion reconstruction result can fall within the theoretical constraint range of both, it indicates that the corrected data conforms to the biological background law of the current sample. In specific operation, the system calculates the stabilization direction residual of the fusion reconstruction result relative to the MG132 perturbation response data, and the decay direction residual of the fusion reconstruction result relative to the corresponding decay trend of the CHX time-series response data. Then, preset penalty weights are assigned to the above bidirectional residuals and combined to construct a negative exponential envelope score, and the following consistency judgment operation is performed:

[0101]

[0102]

[0103]

[0104] in, This represents the stabilization direction residual, used to measure whether the correction result exceeds the theoretical boundary of proteasome inhibition in terms of spatial displacement; This appears and represents the aforementioned newly generated fusion and reconstruction result; This represents the residual in the decay direction, used to measure whether the correction result still follows the natural degradation law in the logarithmic time domain; Represents the natural logarithm function; The decay loss amount generated from the preceding step S2 appears and is invoked; This represents the final generated double residual envelope score, with a value ranging from zero to one. Represents an exponential function; This represents the preset stabilization residual penalty weight; This represents the preset attenuation residual penalty weight, the and All of these are preset weight constants based on fitting the trajectory of historical normal samples.

[0105] Obtaining the double residual envelope score Then, the system executes the final output decision strategy: the system scores the double residual envelope. Consistency threshold with preset (the above) To compare with the empirical lower limit threshold obtained through preliminary experimental data; in the double residual envelope scoring Greater than or equal to the consistency threshold When the system determines that the current fusion reconstruction result has biological consistency, it will combine the fusion reconstruction result (F), the anomaly correction parameter (Q), and the double residual envelope score (F). These data are packaged together and output as the final corrected HSPA8 experimental data; the double residual envelope scoring... Less than the consistency threshold If the system detects an irreversible collapse of the experimental system, it will intercept the output and trigger a reconstruction failure warning.

[0106] During the data reconstruction stage, degradation activity parameters were introduced to recalibrate the potential energy of the MG132 perturbation data. This ensures that the reference value is no longer a simple observation but an upper limit of expression corrected for degradation capacity. Compared to directly using the original MG132 data, this process avoids misleading results due to anomalous accumulation under suppression conditions.

[0107] During the fusion process, anomaly correction parameters are used as weights to perform linear interpolation between the original detection data and the corrected reference values, enabling the results to automatically adjust according to the degree of anomaly. When the degree of anomaly is low, the original data characteristics are preserved; when the degree of anomaly is high, the data converges towards the reference value, thus achieving adaptive correction rather than fixed rule replacement.

[0108] In the consistency assessment stage, the reconstruction results are constrained by constructing and weighting the dual residuals of the stabilization and decay directions. Only data that simultaneously satisfy both biological trends are retained as the final results, thereby avoiding misjudgments caused by a single indicator and improving the biological credibility and consistency of the results.

[0109] This embodiment also provides an automatic correction system for experimental data anomalies related to HSPA8, including:

[0110] The data module acquires HSPA8 protein blot detection data from the same biological sample source, homology perturbation response data based on MG132 and CHX, and Phlda3 association data at the 13891 Da position, and performs normalization processing to obtain a standardized dataset.

[0111] The analysis module performs decoupling analysis on the homogeneous perturbation response data in the standardized dataset to obtain the stabilized release amount and the decay loss amount, and constructs degradation activity parameters that reflect the degradation potential of HSPA8.

[0112] The anomaly correction module performs ratio calculation and deviation assessment on the HSPA8 protein blot detection data and Phlda3 association data in the standardized dataset to obtain the interaction deviation index. It then performs coupling processing on the interaction deviation index in conjunction with the degradation activity parameter to generate anomaly correction parameters.

[0113] The execution module performs weight allocation and fusion reconstruction on the HSPA8 protein blot detection data according to the anomaly correction parameters, and uses the double residual envelope of MG132 and CHX to perform consistency judgment on the fusion reconstruction results, and outputs the corrected HSPA8 experimental data.

[0114] This embodiment also provides a computer device applicable to the automatic correction method for experimental data anomalies related to HSPA8, comprising: a memory and a processor; the memory is used to store computer-executable instructions, and the processor is used to execute the computer-executable instructions to implement the automatic correction method for experimental data anomalies related to HSPA8 as proposed in the above embodiment.

[0115] The computer device can be a terminal, comprising a processor, memory, communication interface, display screen, and input devices connected via a system bus. The processor provides computing and control capabilities. The memory includes non-volatile storage media and internal memory. The non-volatile storage media stores the operating system and computer programs. The internal memory provides an environment for the operation of the operating system and computer programs stored in the non-volatile storage media. The communication interface is used for wired or wireless communication with external terminals; wireless communication can be achieved through Wi-Fi, carrier networks, NFC (Near Field Communication), or other technologies. The display screen can be an LCD screen or an e-ink screen. The input devices can be a touch layer covering the display screen, buttons, a trackball, or a touchpad on the computer device's casing, or an external keyboard, touchpad, or mouse.

[0116] This embodiment also provides a storage medium storing a computer program that, when executed by a processor, implements the automatic correction method for experimental data anomalies related to HSPA8 as proposed in the above embodiments. The storage medium can be implemented by any type of volatile or non-volatile storage device or a combination thereof, such as Static Random Access Memory (SRAM), Electrically Erasable Programmable Read-Only Memory (EEPROM), Erasable Programmable Read-Only Memory (EPROM), Programmable Red-Only Memory (PROM), Read-Only Memory (ROM), magnetic storage, flash memory, magnetic disk, or optical disk.

[0117] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. An automatic correction method for experimental data anomalies related to HSPA8, characterized in that, include: We obtained HSPA8 protein blot detection data, homology perturbation response data based on MG132 and CHX, and Phlda3 association data at the 13891 Da position from the same biological sample, and performed normalization processing to obtain a standardized dataset. Decoupling analysis was performed on the homogeneous perturbation response data in the standardized dataset to obtain the stabilized release amount and the decay loss amount, and a degradation activity parameter reflecting the degradation potential of HSPA8 was constructed. For the HSPA8 protein Western blot detection data and Phlda3 association data in the standardized dataset, a ratio calculation and deviation assessment are performed to obtain the interaction deviation index. The interaction deviation index is then coupled with the degradation activity parameter to generate anomaly correction parameters. Based on the aforementioned anomaly correction parameters, the HSPA8 protein blot detection data are weighted and fused and reconstructed. The double residual envelope of MG132 and CHX is used to perform consistency discrimination on the fusion and reconstruction results, and the corrected HSPA8 experimental data are output.

2. The automatic correction method for experimental data anomalies regarding HSPA8 as described in claim 1, characterized in that: The normalization process to obtain a standardized dataset includes splitting protein lysates from the same biological sample into equal volumes to form a basic detection sample, an MG132-treated sample, and a CHX-treated sample, respectively. Under the same Western blot detection conditions, Western blot imaging was performed on the basic detection sample, the MG132-treated sample, and the CHX-treated sample. The migration interval corresponding to the HSPA8 main band and a preset migration interval centered at 13891 Da were located in each lane, and the integrated grayscale values ​​of the corresponding bands were extracted. At least three background sampling windows far from the band region were selected in each lane to obtain the background grayscale values ​​and the total protein integral value of the corresponding lane. For the CHX-treated sample, Western blot imaging was performed at multiple preset time points, and the integrated grayscale values ​​of the corresponding lanes at each time point were extracted to form CHX time-series response data. Based on the strip integral gray value, background gray value, and total protein integral value, HSPA8 protein blot detection data, MG132 perturbation response data, CHX time series response data, and Phlda3 association data were constructed by combining background gray median subtraction and total protein normalization. The above data were then subjected to uniform standardization processing to obtain standardized data, and a standardized dataset was constructed.

3. The automatic correction method for experimental data anomalies regarding HSPA8 as described in claim 2, characterized in that: The steps of obtaining the stabilized release amount and the decay loss amount include: calculating the relative change rate of the MG132 perturbation response data in the standardized dataset relative to the HSPA8 protein blot detection data, and performing nonlinear logarithmic dimensionality reduction on the relative change rate to extract the stabilized release amount; extracting the logarithmic decay amount of the CHX time-series response data in the standardized dataset at each preset time point relative to the HSPA8 protein blot detection data, assigning progressive weights according to the span ratio of each preset time point in the overall observation time window, and performing time-series accumulation on the weighted logarithmic decay amount to extract the decay loss amount.

4. The automatic correction method for experimental data anomalies regarding HSPA8 as described in claim 3, characterized in that: The degradation activity parameters are constructed by using the stabilized release amount to construct a degradation amplitude correction factor, and by calculating the ratio of the decay loss amount to the degradation amplitude correction factor, performing decoupling and reconstruction processing on the decay loss amount and the stabilized release amount to generate the degradation activity parameters.

5. The automatic correction method for experimental data anomalies regarding HSPA8 as described in claim 4, characterized in that: The method for obtaining the interaction deviation index includes: calculating the relative expression ratio based on the Phlda3 association data corresponding to the basic detection in the standardized dataset and the HSPA8 protein Western blot detection data corresponding to the basic detection. The relative expression ratio is compared with a preset reference ratio center to obtain the deviation; after normalizing the deviation, it is substituted into the hyperbolic tangent function to perform bidirectional soft saturation nonlinear mapping to obtain the interaction deviation index.

6. The automatic correction method for experimental data anomalies regarding HSPA8 as described in claim 5, characterized in that: The generation of anomaly correction parameters includes: assigning preset weight coefficients to the interaction deviation index and the degradation activity parameter respectively and performing a linear combination, substituting the result of the linear combination as a negative exponential term into the logistic regression mapping model to perform probability transformation processing, and using the processing result as the anomaly correction parameters.

7. The automatic correction method for experimental data anomalies regarding HSPA8 as described in claim 6, characterized in that: The weighting and fusion reconstruction of the HSPA8 protein blot detection data includes: using degradation activity parameters to recalibrate the potential energy of the MG132 perturbation response data in the standardized dataset to obtain a corrected reference value; Using the anomaly correction parameter as the fusion weight, linear interpolation fusion was performed between the HSPA8 protein blot detection data and the corrected reference value to obtain the fusion reconstruction result; The consistency judgment of the fusion reconstruction results includes calculating the stabilization direction residual of the fusion reconstruction results relative to the MG132 disturbance response data, and the decay direction residual of the fusion reconstruction results relative to the corresponding decay trend of the CHX time series response data. Preset penalty weights are assigned to the stabilization direction residual and the decay direction residual respectively and combined to obtain the double residual envelope score; the double residual envelope score is compared with a preset consistency threshold, and when the double residual envelope score is greater than or equal to the consistency threshold, the fusion reconstruction result is output as the corrected HSPA8 experimental data. The corrected HSPA8 experimental data includes fusion reconstruction results, anomaly correction parameters, and double residual envelope scores.

8. An automatic data anomaly correction system for HSPA8 experimental data, based on the automatic data anomaly correction method for HSPA8 according to any one of claims 1 to 7, characterized in that: The data module acquires HSPA8 protein blot detection data from the same biological sample source, homology perturbation response data based on MG132 and CHX, and Phlda3 association data at the 13891 Da position, and performs normalization processing to obtain a standardized dataset. The analysis module performs decoupling analysis on the homogeneous perturbation response data in the standardized dataset to obtain the stabilized release amount and the decay loss amount, and constructs degradation activity parameters that reflect the degradation potential of HSPA8. The anomaly correction module performs ratio calculation and deviation assessment on the HSPA8 protein Western blot detection data and Phlda3 association data in the standardized dataset to obtain the interaction deviation index, and performs coupling processing on the interaction deviation index in combination with the degradation activity parameter to generate anomaly correction parameters. The execution module performs weight allocation and fusion reconstruction on the HSPA8 protein blot detection data according to the anomaly correction parameters, and uses the double residual envelope of MG132 and CHX to perform consistency judgment on the fusion reconstruction results, and outputs the corrected HSPA8 experimental data.

9. A computer device comprising a memory and a processor, wherein the memory stores a computer program, characterized in that: When the processor executes the computer program, it implements the steps of the automatic correction method for experimental data anomalies related to HSPA8 as described in any one of claims 1 to 7.

10. A computer-readable storage medium having a computer program stored thereon, characterized in that: When the computer program is executed by the processor, it implements the steps of the automatic correction method for experimental data anomalies related to HSPA8 as described in any one of claims 1 to 7.

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