Gut microbiota-lncRNA/circRNA crosstalk: implications for different diseases.

The gut microbiota features an abundance of diverse microorganisms and represents an important component of human physiology and metabolic homeostasis, indicating their roles in a wide array of physiological and pathological processes in the host. Maintaining balance in the gut microbiota is critical for normal functionality as microbial dysbiosis can lead to the occurrence and development of diseases through various mechanisms. Long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs) are non-coding RNAs that perform important regulatory functions for many processes. Furthermore, the gut microbiota and lncRNAs/circRNAs are known to interact in a range of both physiological and pathological activities. In this article, we review existing research relevant to the interaction between the gut microbiota and lncRNAs/circRNAs and investigate the role of their crosstalk in the pathogenesis of different diseases. Studies have shown that, the gut microbiota can target lncRNAs ENO1-IT1, BFAL1, and LINC00152 to regulate colorectal cancer development via various signaling pathways. In addition, the gut microbiota can influence mental diseases and lung tumor metastasis by modulating circRNAs such as circNF1-419, circ_0001239, circHIPK2 and mmu_circ_0000730. These findings provide a theoretical basis for disease prevention and treatment and suggest that gut microbiota-lncRNA/circRNA crosstalk has high clinical value.

Component analysis using UPLC-Q-TOF/MS and quality evaluation using fingerprinting and chemometrics for hops.

Hops, extensively cultivated in China for their food and medicinal applications, currently lack well-defined chemical markers to evaluate variations in their quality. The study aimed to explore variations in the quality of Chinese hops by the chemical characteristics of hops, employing UPLC-Q-TOF/MS, integrated with chemical fingerprinting and chemometrics. The results indicated that Chinese hops are abundant in polyphenols and bitter acids. The integration of UPLC-Q-TOF/MS, Chemical fingerprinting, and chemometrics revealed to be an accurate and effective approach for assessing the quality of Chinese hops. In this study, ten important chemical markers were found to be useful in differentiating various hop varieties. Moreover, the support vector machine showed a prediction accuracy of 92.3077% in identifying Chinese hop varieties. The strategy of the study lays the groundwork for classifying Chinese hop varieties and serves as a prerequisite for future quality control studies, particularly focusing on chemical compositions.

The Role of Circular RNA Variants Generated from the NFIX Gene in Different Diseases.

Circular RNAs (circRNAs) have been identified as important regulators in different developmental processes and disease pathogenesis. The loop structure of circRNAs makes them very stable in different conditions and microenvironments. circRNAs can affect microRNA (miRNA) and RNA binding protein (RBP) activity, encode functional proteins and regulate gene transcription. Recently, two circNFIX variants derived from the same gene, the Nuclear Factor I X (NFIX) gene, were determined as participants in the pathological processes of various diseases such as heart diseases and cancers. Both circNFIX variants are exonic circular RNAs and mainly function by sponging miRNAs. In this review, we summarize the current knowledge on circRNAs, elucidate the origins and properties of two circNFIX variants, explore the roles of two circNFIX variants in different diseases, and present clinical perspectives.

The important regulatory roles of circRNA­encoded proteins or peptides in cancer pathogenesis (Review).

Circular RNAs (circRNAs) represent a class of RNA molecules characterized by their covalently closed structures. There are three types of circRNAs, namely exonic circRNAs, exon­intron circRNAs and circular intronic RNAs. To date, four distinct mechanisms have been unveiled through which circRNAs exert their functional influence, including serving as microRNA (miRNA) sponges, interacting with RNA binding proteins (RBPs), modulating parental gene transcription and acting as templates for translation. Of note, among these mechanisms, the miRNA/RBP sponge function has been the most investigated one. Recent research has uncovered the presence of various proteins or peptides encoded by circRNA. CircRNAs are translated independent of the 5' cap and 3' polyA tail, which are typical elements for linear RNA translation. Some unique elements, such as internal ribosome entry sites and N­methyladenosine modifications, facilitate the initiation of translation. These circRNA­encoded proteins or peptides participate in diverse signalling pathways and act as important regulators in carcinogenesis by influencing cell proliferation, migration, apoptosis and other key processes. Consequently, circRNA­encoded proteins or peptides have great potential as therapeutic targets for anticancer drugs. The present comprehensive review aimed to systematically summarize the current understanding of circRNA­encoded proteins or peptides and to unveil their roles in carcinogenesis.

Underlying mechanisms and clinical potential of circRNAs in glioblastoma.

Glioblastoma (GBM) is the most malignant form of glioma and is difficult to diagnose, leading to high mortality rates. Circular RNAs (circRNAs) are noncoding RNAs with a covalently closed loop structure. CircRNAs are involved in various pathological processes and have been revealed to be important regulators of GBM pathogenesis. CircRNAs exert their biological effects by 4 different mechanisms: serving as sponges of microRNAs (miRNAs), serving as sponges of RNA binding proteins (RBPs), modulating parental gene transcription, and encoding functional proteins. Among the 4 mechanisms, sponging miRNAs is predominant. Their good stability, broad distribution and high specificity make circRNAs promising biomarkers for GBM diagnosis. In this paper, we summarized the current understanding of the characteristics and action mechanisms of circRNAs, illustrated the underlying regulatory mechanisms of circRNAs in GBM progression and explored the possible diagnostic role of circRNAs in GBM.

CircRNA-miRNA-VEGFA: an important pathway to regulate cancer pathogenesis.

Cancers, especially malignant tumors, contribute to high global mortality rates, resulting in great economic burden to society. Many factors are associated with cancer pathogenesis, including vascular endothelial growth factor-A (VEGFA) and circular RNAs (circRNA). VEGFA is a pivotal regulator of vascular development such as angiogenesis, which is an important process in cancer development. CircRNAs have covalently closed structures, making them highly stable. CircRNAs are widely distributed and participate in many physiological and pathological processes, including modulating cancer pathogenesis. CircRNAs act as transcriptional regulators of parental genes, microRNA (miRNA)/RNA binding protein (RBP) sponges, protein templates. CircRNAs mainly function via binding to miRNAs. CircRNAs have been shown to influence different diseases such as coronary artery diseases and cancers by regulating VEGFA levels via binding to miRNAs. In this paper, we explored the origin and functional pathways of VEGFA, reviewed the current understanding of circRNA properties and action mechanisms, and summarized the role of circRNAs in regulating VEGFA during cancer pathogenesis.

HaASR2 from Haloxylon ammodendron confers drought and salt tolerance in plants.

Abscisic acid (ABA), stress, and ripening-induced proteins (ASR), which belong to the ABA/WDS domain superfamily, are involved in the plant response to abiotic stresses. Haloxylon ammodendron is a succulent xerohalophyte species that exhibits strong resistance to abiotic stress. In this study, we isolated HaASR2 from H. ammodendron and demonstrated its detailed molecular function for drought and salt stress tolerance. HaASR2 interacted with the HaNHX1 protein, and its expression was significantly up-regulated under osmotic stress. Overexpression of HaASR2 improved drought and salt tolerance by enhancing water use efficiency and photosynthetic capacity in Arabidopsis thaliana. Overexpression of HaASR2 maintained the homeostasis of reactive oxygen species (ROS) and decreased sensitivity to exogenous ABA and endogenous ABA levels by down-regulating ABA biosynthesis genes under drought stress. Furthermore, a transcriptomic comparison between wild-type and HaASR2 transgenic Arabidopsis plants indicated that HaASR2 significantly induced the expression of 896 genes in roots and 406 genes in shoots under osmotic stress. Gene ontology (GO) enrichment analysis showed that those DEGs were mainly involved in ROS scavenging, metal ion homeostasis, response to hormone stimulus, etc. The results demonstrated that HaASR2 from the desert shrub, H. ammodendron, plays a critical role in plant adaptation to drought and salt stress and could be a promising gene for the genetic improvement of crop abiotic stress tolerance.

Analysis of the relationship between axial length, optic disc morphology, and regional variations in retinal vessel density in young adults with healthy eyes.

Purpose: To investigate the relationship between optic disc morphology, axial length, and regional distribution of retinal vessels in healthy eyes of young adults. Methods: Nine hundred and two healthy eyes were enrolled in this university-based, cross-sectional study. Spectral-domain optical coherence tomography angiography was used to measure the parapapillary retinal vessel density. We automated the process of calculating optic disc tilt and rotation by using a program written in Python. Relationships between optic disc rotation, optic disc tilt, parapapillary vessel density, and other ocular parameters were analyzed using regression models. Results: As axial length increased, optic disc morphology became more tilted and rotated inferiorly. The superficial vessel density (SVD) and radial peripapillary capillary density (RPCD) gradually decreased in all regions except for the temporal quadrant. Increased temporal SVD (OR [95% CI] = 1.081 [1.039, 1.124], p < 0.001), reduced nasal SVD (OR [95% CI] = 0.898 [0.861, 0.937], p < 0.01), and short relative lens position (OR [95% CI] = 0.126 [0.032, 0.495], p = 0.003) were significantly associated with the presence of a tilted optic disc. Inferior disc rotation was associated with decreased superior deep vessel density (DVD) and increased inferior DVD and temporal DVD after adjusting for sex and axial length. Conclusion: The tilted and rotated optic discs were associated with the distribution of SVD and DVD, respectively. We should fully consider the influence of optic disc morphology on parapapillary vessel density in eyes with myopia.

Potential of curcumin loaded nanoparticles in ovarian cancer: Investigation using gynecological color Doppler ultrasound technique.

Doppler ultrasonography is a type of medical ultrasonography that uses the Doppler effect to provide images of the movement of tissues and bodily fluids (typically blood) relative to the probe. To determine the potential of curcumin loaded nanoparticles in ovarian cancer which was diagnosed by using the gynecological color doppler ultrasound technique. Curcumin (CRMN) loaded chitosan nanoparticles were formulated using the ionotropic gelation method and characterized for particle size, zeta potential and polydispersity index (PDI). Clinical parameters like serum creatinine, blood serum urea nitrogen, resistance index and peak systolic velocity were evaluated. The drug loading efficiency was found between 11.38 to 17.45% with a particle size of 140-220nm. The zeta potential ranged between 19.12 to 23.14mV. Clinical parameters were found significantly changed when compared with before injection of CRMN-loaded nanoparticles. BUN was increased from 7.11±0.25to 28.27±6.65 mmol/L while SCr was also found to be increased from 52.71±3.14 µmol/L to 312.20±40.41 µmol/L. Collectively, these images of color doppler in animal model demonstrated efficient use in the diagnosis of ovarian cancer. This study confirms the potential of color doppler as an efficient medical imaging tool for ovarian cancer.

The Functional Roles of Lactobacillus acidophilus in Different Physiological and Pathological Processes.

Probiotics are live microorganisms that can be consumed by humans in amounts sufficient to offer health-promoting effects. Owing to their various biological functions, probiotics are widely used in biological engineering, industry and agriculture, food safety, and the life and health fields. Lactobacillus acidophilus (L. acidophilus), an important human intestinal probiotic, was originally isolated from the human gastrointestinal tract and its functions have been widely studied ever since it was named in 1900. L. acidophilus has been found to play important roles in many aspects of human health. Due to its good resistance against acid and bile salts, it has broad application prospects in functional, edible probiotic preparations. In this review, we explore the basic characteristics and biological functions of L. acidophilus based on the research progress made thus far worldwide. Various problems to be solved regarding the applications of probiotic products and their future development are also discussed.

The Functional Roles of Lactobacillus acidophilus in Different Physiological and Pathological Processes.

Probiotics are live microorganisms that can be consumed by humans in amounts sufficient to offer health-promoting effects. Owing to their various biological functions, probiotics are widely used in biological engineering, industry and agriculture, food safety, and the life and health fields. Lactobacillus acidophilus (L. acidophilus), an important human intestinal probiotic, was originally isolated from the human gastrointestinal tract and its functions have been widely studied ever since it was named in 1900. L. acidophilus has been found to play important roles in many aspects of human health. Due to its good resistance against acid and bile salts, it has broad application prospects in functional, edible probiotic preparations. In this review, we explore the basic characteristics and biological functions of L. acidophilus based on the research progress made thus far worldwide. Various problems to be solved regarding the applications of probiotic products and their future development are also discussed.

The circRNA-miRNA/RBP regulatory network in myocardial infarction.

Myocardial infarction (MI) is a serious heart disease that causes high mortality rate worldwide. Noncoding RNAs are widely involved in the pathogenesis of MI. Circular RNAs (circRNAs) are recently validated to be crucial modulators of MI. CircRNAs are circularized RNAs with covalently closed loops, which make them stable under various conditions. CircRNAs can function by different mechanisms, such as serving as sponges of microRNAs (miRNAs) and RNA-binding proteins (RBPs), regulating mRNA transcription, and encoding peptides. Among these mechanisms, sponging miRNAs/RBPs is the main pathway. In this paper, we systematically review the current knowledge on the properties and action modes of circRNAs, elaborate on the roles of the circRNA-miRNA/RBP network in MI, and explore the value of circRNAs in MI diagnosis and clinical therapies. CircRNAs are widely involved in MI. CircRNAs have many advantages, such as stability, specificity, and wide distribution, which imply that circRNAs have a great potential to act as biomarkers for MI diagnosis and prognosis.

Prevalence and inconformity of refractive errors and ocular biometry of 3573 medical university freshman students for 4 consecutive years.

AIM: To evaluate the prevalence of refractive errors and ocular biometry in 3573 freshman students at Tianjin Medical University for 4 consecutive years. METHODS: In this university-based, cross-sectional study, comprising 3573 students, visual acuity (VA), slit-lamp examination, non-cycloplegic auto-refraction, and ocular biometry were recorded. RESULTS: The prevalence of myopia increased annually, from 2017 to 2020 were 93.5%, 94.5%, 95.9%, and 96.2%, respectively (P=0.03), and the prevalence of high myopia was 25.7%, 26.9%, 28.6%, and 28.6%, respectively. Males tended to have a higher percentage of total astigmatism than females, with astigmatism ≥0.75 and ≥1.0 D criteria. The percentage of with-the-rule astigmatism, against-the-rule astigmatism, and oblique astigmatism was 90.3%, 5.8%, and 3.9%, respectively, with astigmatism ≥1.00 D criteria. The mean spherical equivalent, axial length (AL), central corneal thickness (CCT), anterior chamber depth (ACD), lens thickness (LT), corneal radius (CR), and lens position (LP) were 4.37±2.52 D, 25.28±1.24 mm, 539.49±34.98 µm, 3.31±0.34 mm, 3.47±0.21 mm, 7.8±0.28 mm, and 5.04±0.32 mm, respectively. With diopter increase in myopia, the AL became longer, CR became steeper, ACD became deeper, LT became thinner, and LP became more posterior (all P<0.01). Females had a shorter AL, thinner CCT, smaller CR, shallower ACD, thicker lens, and more anterior LP than males (P<0.01). The 64% of high myopia had AL≥26 mm, meanwhile, 5.8% mild myopia and 21.1% moderate myopia had AL≥26 mm. With AL≥26 mm, mild and moderate myopia compared to high myopia, AL was shorter (26.51±0.46 vs 26.87±0.70 mm), CR was larger (8.10±0.3 vs 7.85±0.23 mm) and LT was thinner (3.39±0.19 vs 3.45±0.19 mm, P<0.001). CONCLUSION: The prevalence of myopia and high myopia is significantly high in freshman students. The majority of astigmatism is with-the-rule. Inconformity of refractive errors and ocular biometry existed in some students. Attention should be paid to the ocular biometry of myopia.

Maslinic acid activates renal AMPK/SIRT1 signaling pathway and protects against diabetic nephropathy in mice.

BACKGROUND: Diabetic nephropathy has been a devastating complication. Clinically, there is an urgent need for nephroprotective agents to delay the onset of diabetic nephropathy and ameliorate its symptoms. Maslinic acid is a pentacyclic triterpene acid with protective effect on multiple organs against oxidative stress and inflammation. In this research, we hypothesized that maslinic acid protects renal function against diabetic nephropathy. METHODS: C57BL/6 J male mice administrated with 50 mg/kg of Streptozocin (STZ) daily were used to establish diabetic mouse model (blood glucose levels > 300 mg/dL). Urinary levels of albumin, total proteins, and creatinine were analyzed by an automatic analyzer. H&E staining was used to evaluate renal damage. qRT-PCR and ELISA were performed to investigate the inflammation and oxidative stress in renal tissues. Western blot was used to assess the activation of AMPK signaling. RESULTS: Maslinic acid treatment alleviated the loss of body weight and blood glucose in diabetic mice. Renal structure and function were protected by maslinic acid in diabetic mice. 20 mg/kg maslinic acid treatment for 8 weeks significantly alleviated the oxidative stress and inflammation in the kidney of diabetic rats. Maslinic acid treatment activated the renal AMPK/SIRT1 signaling pathway. CONCLUSION: Maslinic acid ameliorates diabetic nephropathy and activates the renal AMPK/SIRT1 signaling pathway.

Impact of a community-based health education intervention on awareness of influenza, pneumonia, and vaccination intention in chronic patients.

The study aimed to examine the impact of a community-based health education intervention on the awareness and attitude of influenza and Streptococcus pneumoniae vaccination in patients with chronic diseases. We selected study participants from two counties in Jiaxing City, Zhejiang Province. We conducted a household baseline survey with quarterly follow-up and health education for all chronic disease patients in the area. A total of 720 patients with chronic diseases were included in two rounds of questionnaire surveys before and after a year-long health education related to the influenza and Streptococcus pneumoniae vaccination. Before the community education, 59.9% and 59.6% of study participants self-report the awareness of the hazards of influenza and pneumonia, respectively; while only 17.7% and 6.0% study participants self-reported awareness of the influenza or pneumonia vaccination, respectively; the self-reported vaccination rates were 1.3% and 0% for influenza and pneumonia, respectively. One year after the intervention, all rates were increased significantly, but the influenza vaccination rate was still far below the WHO minimum target level (75%). The main reason for not getting vaccinated reported by study participants was that participants thought they were in good health and would not become sick easily. There was no statistically significant difference between the impacts of health care practitioners' and general practitioners' recommendations for vaccination. Continued and pertinent health education has a significant impact on influenza and pneumonia vaccination in patients with chronic diseases.

Relationship Between Myopia and Other Risk Factors With Anxiety and Depression Among Chinese University Freshmen During the COVID-19 Pandemic.

Purpose: To investigate the association of myopia and other risk factors with anxiety and depression among Chinese university freshmen during the coronavirus disease 2019 (COVID-19) pandemic. Methods: This cross-sectional study was conducted at the Tianjin Medical University from October 2020 to December 2020. Ophthalmic examination of the eyes was performed by an experienced ophthalmologist. Detailed information on depression, anxiety, and other risk factors was collected via the Self-rating Anxiety Scale and Self-rating Depression Scale. Results: The overall prevalence of anxiety and depression in our study was 10.34 and 25.13%, respectively. The prevalence of myopia and high myopia as 92.02 and 26.7%, respectively. There were significant associations between anxiety and spectacle power [odds ratios (OR) = 0.89; 95% CI: 0.81-0.98, P = 0.019], sphere equivalent (OR = 0.89; 95% CI: 0.81- 0.98, P = 0.025), sleep time (OR = 0.53; 95% CI: 0.35-0.79, P = 0.002), and body mass index (OR = 0.93; 95% CI: 0.86-0.99, P = 0.047). In the multivariable linear regression models, spectacle power (ß = -0.43; 95% CI: -0.68 to -0.19, P = 0.001) and sphere equivalent (ß = -0.36; 95% CI: -0.60 to -0.11, P = 0.005) were negatively associated with anxiety scores, whereas axial length (ß = 0.54; 95% CI: 0.02-1.07, P = 0.044) was positively correlated with anxiety scores. Every 1 h decrease in sleep time was associated with a 0.12-point increase in depression score. Conclusion: Myopia was associated with anxiety and anxiety scores. The greater the degree of myopia, the higher the anxiety score. However, myopia was not found to be associated with depression. The results highlight the importance of providing psychological support to students with myopia during the COVID-19 pandemic.

CircHIPK3 Plays Vital Roles in Cardiovascular Disease.

Circular RNAs (circRNAs) are covalently closed RNAs that function in various physiological and pathological processes. CircRNAs are widely involved in the development of cardiovascular disease (CVD), one of the leading causes of morbidity and mortality worldwide. CircHIPK3 is generated from the second exon of the HIPK3 gene, a corepressor of homeodomain transcription factors. As an exonic circRNA (ecRNA), circHIPK3 is produced through intron-pairing driven circularization facilitated by Alu elements. In the past 5 years, a growing number of studies have revealed the multifunctional roles of circHIPK3 in different diseases, such as cancer and CVD. CircHIPK3 mainly participates in CVD pathogenesis through interacting with miRNAs. This paper summarizes the current literature on the biogenesis and functions of circHIPK3, elucidates the role of circHIPK3 in different CVD patterns, and explores future perspectives.

Endometrial thickness in the prediction of neonatal adverse outcomes in frozen cycles for singleton pregnancies.

RESEARCH QUESTION: Does endometrial thickness (EMT) predict adverse neonatal outcomes in singleton pregnancies after in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) frozen embryo transfer (FET)? DESIGN: This retrospective study involved 13,383 women undergoing IVF/ICSI FET cycles between January 2010 and December 2018 in Women's Hospital of Zhejiang University. The primary outcome was preterm delivery (PTD). The secondary outcomes were small for gestational age (SGA), large for gestational age (LGA) and low birthweight (LBW). RESULTS: A total of 13,383 FET cycles resulting in 5220 singleton live births and 8163 failed cycles were included. Multiple spline regression visualization showed an increasing risk of PTD and LBW for a thin EMT. By comparing multiple cut-off points using area under the curve, a cut-off point of 8 mm was identified, which was used to categorize EMT. A reference point of EMT greater than 8 mm was used; after adjusting for covariates, individuals with EMT less than 8 mm had an adjusted odds ratio of 1.75 (95% CI 1.30 to 2.34) for PTD, 1.57 (95% CI 1.09 to 2.26) for LBW, 0.97 (95% CI 0.63 to 1.50) for SGA and 1.04 (95% CI 0.79 to 1.37) for LGA. Additional analyses showed similar increasing risk with a thin endometrium for both PTD with and without caesarean section, and PTD with low and normal birthweight percentiles. CONCLUSION: A clinical cut-off point of 8 mm has been identified, below which risk of PTD and LBW increases in women undergoing IVF/ICSI.

Computed Tomography Image Analysis and Clinical Diagnosis Value of Nasal Olfactory Neuroblastoma.

BACKGROUND: This article retrospectively analyzed the data of all patients diagnosed with malignant tumors of the nasal cavity and sinuses by pathologic section in all patients (including outpatients and inpatients) in our hospital from January 2008 to December 2017 (10 years) to discuss the nasal cavity The clinical characteristics of sinus malignant tumors and the clinical characteristics (symptoms, signs, imaging examinations) of 35 cases of olfactory neuroblastoma with high incidence were analyzed, and their treatment methods and prognosis were summarized for future nasal tumors. CLINICAL BASIS FOR PREVENTION AND TREATMENT: In our department, 300 cases of malignant tumors of nasal cavity and sinuses were treated within 10 years. AGE: Eighty-six cases (28.6%) were in the age group 50-59 years old, occupying first place, and 55 cases (18.3%) were in the age group 60-69 years old, occupying second place. By comparing groups A and B with malignant tumors of the nasal cavity and sinuses, the difference in age distribution of olfactory neuroblastoma is statistically significant. Of the 35 cases of olfactory neuroblastoma, 23 cases were diagnosed with nasal tumors on admission, 12 cases were initially diagnosed with nasal polyps (including 7 cases with hemorrhagic and necrotic nasal polyps), and 20 cases underwent rapid pathologic definite diagnosis during the operation. DIAGNOSIS FOR POSTOPERATIVE PATHOLOGIC RETURNS: On the basis of our analysis, the following conclusions can be drawn: Of the 300 cases of malignant tumors of the nasal cavity and paranasal sinuses, age distribution tends to be younger, the overall incidence of males is higher than that of females, but the proportion of male and female olfactory neuroblastoma is equal, and the incidence of adenoid cystic carcinoma and sarcoma. The morphology of cancer can be divided as follows: the tissue source is epithelial tissue, the cancer is harder, paler, drier, and often forms cancer nests, the parenchymal and interstitial boundaries are clear, and the fibrous tissue is hyperplastic. The incidence rate of females is higher than that of males, with the primary origin in the sinuses. Among them, the maxillary sinus is more common. Olfactory neuroblastoma, lymphoma, and sarcoma are gradually increasing in malignant tumors of the nasal cavity and sinuses.

Vulnerability Mining Method for the Modbus TCP Using an Anti-Sample Fuzzer.

Vulnerability mining technology is used for protecting the security of industrial control systems and their network protocols. Traditionally, vulnerability mining methods have the shortcomings of poor vulnerability mining ability and low reception rate. In this study, a test case generation model for vulnerability mining of the Modbus TCP based on an anti-sample algorithm is proposed. Firstly, a recurrent neural network is trained to learn the semantics of the protocol data unit. The softmax function is used to express the probability distribution of data values. Next, the random variable threshold and the maximum probability are compared in the algorithm to determine whether to replace the current data value with the minimum probability data value. Finally, the Modbus application protocol (MBAP) header is completed according to the protocol specification. Experiments using the anti-sample fuzzer show that it not only improves the reception rate of test cases and the ability to exploit vulnerabilities, but also detects vulnerabilities of industrial control protocols more quickly.