Development and evaluation of a program based on a generative pre-trained transformer model from a public natural language processing platform for efficiency enhancement in post-procedural quality control of esophageal endoscopic submucosal dissection.

BACKGROUND: Post-procedural quality control of endoscopic submucosal dissection (ESD) is emphasized in guidelines. However, this process can be tedious and time-consuming. Recently, a pre-training model called generative pre-trained transformer (GPT) on a public natural language processing platform has emerged and garnered significant attention, whose capabilities align well with the post-procedural quality control process and have the potential to streamline it. Therefore, we developed a simple program utilizing this platform and evaluated its performance. METHODS: Esophageal ESDs were retrospectively included. The manual quality control process was performed and act as reference standard. GPT's prompt was optimized through multiple iterations. A Python program was developed to automatically submit prompt with pathological report of each ESD procedure and collect quality control information provided by GPT. Its performance on quality control was evaluated with accuracy, precision, recall, and F-1 score. RESULTS: 165 cases were involved into the dataset, of which 5 were utilized as the prompt optimization dataset and 160 as the validation dataset. Definitive prompt was achieved through seven iterations. Time spent on the validation dataset by GPT was 13.47 ± 2.43 min. Accuracies of pathological diagnosis, invasion depth, horizontal margin, vertical margin, vascular invasion, and lymphatic invasion of the quality control program were (0.940, 0.952) (95% CI), (0.925, 0.945) (95% CI), 0.931, 1.0, and 1.0, respectively. Precisions were (0.965, 0.969) (95% CI), (0.934, 0.954) (95% CI), and 0.957 for pathological diagnosis, invasion depth, and horizontal margin, respectively. Recalls were (0.940, 0.952) (95% CI), (0.925, 0.945) (95% CI), and 0.931 for factors as mentioned, respectively. F1-score were (0.945, 0.957) (95% CI), (0.928, 0.948) (95% CI), and 0.941 for factors as mentioned, respectively. CONCLUSIONS: This quality control program was qualified of post-procedural quality control of esophageal ESDs. GPT can be easily applied to this quality control process and reduce workload of the endoscopists.

Comparison of efficiency and safety between dual-clip and rubber band-assisted ESD and conventional ESD for colonic lateral spreading tumors (LSTs) with different levels of technical difficulty: a retrospective case-control study.

BACKGROUND: Dual-clip and rubber band-assisted endoscopic submucosal dissection (DCRB-ESD) is a useful technique in the management of lateral spreading tumors (LSTs) of the colon and is suggested by researchers compared with conventional ESD (C-ESD). The aim of this retrospective study is to further analyze the efficiency and safety of DCRB-ESD in a setting with varying technical difficulties. METHODS: Patients who underwent endoscopic treatment (DCRB-ESD or C-ESD) due to LSTs between Jan 1st, 2019 and Jan 1st, 2022, were retrospectively collected. Patients were classified into the following two groups: the DCRB-ESD group (n = 46) and the C-ESD group (n = 81). Baselines were compared and propensity score matching (PSM) was employed to manage the heterogeneity. The technical difficulty and outcomes of the two groups were evaluated based on a semiquantitative model (CS-CRESD) previously described. RESULTS: The baseline characteristics of the two groups were balanced except sex and LST classification before PSM and were corrected after PSM. The median ESD operation time of DCRB-ESD was shorter than that of C-ESD (32 vs 41 and 30 vs 44 before and after PSM respectively, P < 0.05). The operation durations of cases with different CS-CRESD scores were different (P < 0.05). In the subgroup with a score of 0, DCRB-ESD showed no advantage than C-ESD in terms of operation duration before and after PSM. In subgroups with a score of 1-3, DCRB-ESD was faster than C-ESD. In subgroups with a score of 4-5, the between-group operation duration was not significantly different due to the limited number of cases, although the median time of DCRB-ESD was shorter. The R0 resection rates, curative resection, complications, and additional surgery in both groups were not significantly different. No adverse events, such as a clip falling off or rubber band rupturing occurred during this study. CONCLUSION: DCRB-ESD was an efficient and safe procedure in the management of colonic LSTs. With DCRB-ESD, the operation duration of difficult cases can be shortened without sacrificing complication risk. However, not all cases would benefit from DCRB-ESD. For easy cases (CS-CRESD score = 0), DCRB-ESD may not be prior to C-ESD by experienced endoscopists. A pre-ESD technical difficulty evaluation was recommended to decide whether to perform DCRB-ESD or not.

Development of a droplet digital PCR assay to detect bovine alphaherpesvirus 1 in bovine semen.

BACKGROUND: Infectious bovine rhinotracheitis (IBR) caused by bovine alphaherpesvirus 1 (BoHV-1) is one of the most important contagious diseases in bovine. This is one of the most common infectious disease of cattle. This has led to high economic losses in the cattle farming industry. BoHV-1 can potentially be transmitted via semen during natural or artificial insemination (AI). Therefore, testing methods for the early diagnosis of BoHV-1 infection are urgently needed for international trade of ruminant semen. In this study, we developed a novel droplet digital PCR (ddPCR) assay for the detection of BoHV-1 DNA in semen samples. RESULTS: The ddPCR results showed that the detection limit was 4.45 copies per reaction with high reproducibility. The established method was highly specific for BoHV-1 and did not show cross-reactivity with specify the organisms (BTV, BVDV, Brucella, M . bovis). The results of clinical sample testing showed that the positivity rate of ddPCR (87.8%) was higher than that of qPCR (84.1%). CONCLUSIONS: The ddPCR assay showed good accuracy for mixed samples and could be a new added diagnostic tool for detecting BoHV-1.

ERK1/2 mitogen-activated protein kinase mediates downregulation of intestinal tight junction proteins in heat stress-induced IBD model in pig.

In many mammalian species, including pigs, heat stress (HS) detrimentally leads to epithelium damage and increases intestinal permeability. However, the underlying molecular mechanisms are not thoroughly investigated yet. This study aimed to examine the RIP1/RIP3-ERK1/2 signaling pathway that regulates the expression of tight junction proteins in HS-treated pigs. In in vitro cultured intestinal porcine epithelial cells (IPEC-J2), HS induced the expression of tight junction proteins, ZO-1, claudin-1, and claudin-4, that are regulated by the ERK1/2-MAPK signaling pathway. Further, high expression of HSP70 in IPEC-J2 cells induced a significant decrease in receptor-interacting protein 1/3 (RIP1/3), phosphorylated ERK, and tight junction protein claudin-1 (P < 0.05). Necrostatin-1 (A selective inhibitor of RIPK1) suppressed the upregulation of phosphorylated ERK1/2 induced by HS, indicating that the RIP1/RIP3 regulates ERK1/2 phosphorylation in IPEC-J2 under heat stress. In addition, HS significantly damaged the intestinal morphology characterized by reduction of villus length and crypt depth in in vivo porcine model. Moreover, the expression of tight junction, ZO-1, and claudin-4 were downregulated, whereas phosphorylated p38 and ERK1/2 were upregulated in the duodenum of heat-stressed pigs. Interestingly, a decrease in ZO-1 and claudin-1 was observed in the colon, where phosphorylated ERK1/2 was similar to that in the duodenum. Our results demonstrate that RIP1/RIP3-ERK1/2 signaling pathway regulates the expression of tight junction proteins in HS-pigs. This finding further advances the intestinal barrier function's underlying mechanisms associated with signaling regulation.

Dynamics of apoptosis-related gene expression during follicular development in yak.

The potential reproduction power of domestic animals is limited by a complicated follicular atresia process. P53, caspase-9 (Casp9), Bax, Bcl-2 and Fas play a crucial role in the ovarian mitochondrion-dependent apoptosis and death receptor pathway. In accordance with this study, the expression levels of Casp9, Bax, Bcl-2 and Fas were analysed in ovaries and oviducts of yak by immunohistochemistry (IHC). P53 and the above in ovarian granulosa cells (GCs) from atretic (3-6 mm) to healthy follicles (6-8 mm) and in oviducts were examined from the luteal phase to the follicular phase during the oestrous circle by Western blot (WB) and real-time PCR (RT-PCR). Results demonstrated that typical classic apoptotic factors Casp9, Bax, Bcl-2 and Fas were expressed in the cytoplasm and zonal pellucida of oocytes, primordial follicles, primary follicles, ovarian surface epithelium, ovarian GCs, granular lutein cells, surface epithelia in oviduct uterotubal junction and oviduct ampulla during the luteal phase. RT-PCR and WB revealed that P53 and Fas significantly increased in GCs of atretic follicles. P53 and Casp9 increased in oviduct epithelium during the luteal phase, but Fas was unchanged. A contrary tendency was noted in Bcl-2 and Bax expression. Overall, P53 and Fas play an essential role in inducing GC apoptosis, and Bax, Bcl-2, Casp9 and P53 are involved in oviduct epithelial regeneration in yak.

Delayed bowel obstruction after seat belt injury: a case report.

BACKGROUND: Delayed bowel obstruction due to seat belt injury is extremely rare. The delayed onset of nonspecific symptoms makes a timely diagnosis difficult. A deep understanding of the characteristics of this condition is helpful for early diagnosis and treatment. CASE PRESENTATION: A 39-year-old male was transferred to our hospital from another hospital complaints of progressive abdominal distension and severe weakness. In the previous hospital, he was diagnosed with "adult megacolon" and was recommended for surgical treatment. In our hospital, he was diagnosed with delayed bowel obstruction due to seat belt injury and underwent surgical intervention. Following laparoscopic adhesiolysis and resection of the narrow small intestine, his symptoms improved rapidly, and he was discharged. CONCLUSION: Delayed bowel obstruction due to seat belt injury may present clinical symptoms any time after the injury. Imaging examination, ileus tube and small colonoscopy may provide us with valuable cues for the diagnosis and treatment of delayed bowel obstruction, and laparoscopy may be an alternative approach in surgical intervention.

Proteomic study of hypothalamus in pigs exposed to heat stress.

BACKGROUND: With evidence of warming climates, it is important to understand the effects of heat stress in farm animals in order to minimize production losses. Studying the changes in the brain proteome induced by heat stress may aid in understanding how heat stress affects brain function. The hypothalamus is a critical region in the brain that controls the pituitary gland, which is responsible for the secretion of several important hormones. In this study, we examined the hypothalamic protein profile of 10 pigs (15 ± 1 kg body weight), with five subjected to heat stress (35 ± 1 °C; relative humidity = 90%) and five acting as controls (28 ± 3 °C; RH = 90%). RESULT: The isobaric tags for relative and absolute quantification (iTRAQ) analysis of the hypothalamus identified 1710 peptides corresponding to 360 proteins, including 295 differentially expressed proteins (DEPs), 148 of which were up-regulated and 147 down-regulated, in heat-stressed animals. The Ingenuity Pathway Analysis (IPA) software predicted 30 canonical pathways, four functional groups, and four regulatory networks of interest. The DEPs were mainly concentrated in the cytoskeleton of the pig hypothalamus during heat stress. CONCLUSIONS: In this study, heat stress significantly increased the body temperature and reduced daily gain of body weight in pigs. Furthermore, we identified 295 differentially expressed proteins, 147 of which were down-regulated and 148 up-regulated in hypothalamus of heat stressed pigs. The IPA showed that the DEPs identified in the study are involved in cell death and survival, cellular assembly and organization, and cellular function and maintenance, in relation to neurological disease, metabolic disease, immunological disease, inflammatory disease, and inflammatory response. We hypothesize that a malfunction of the hypothalamus may destroy the host physical and immune function, resulting in decreased growth performance and immunosuppression in heat stressed pigs.

Magnetic molecularly imprinted specific solid-phase extraction for determination of dihydroquercetin from Larix griffithiana using HPLC.

The naturally occurring quercetin flavonoid, dihydroquercetin, is widely distributed in plant tissues and has a variety of biological activities. Herein, a magnetic molecularly imprinted solid-phase extraction was tailor made for selective determination of dihydroquercetin in Larix griffithiana using high-performance liquid chromatography. Amino-functionalized core-shell magnetic nanoparticles were prepared and characterized using scanning electron microscopy, transmission electron microscopy, vibrating sample magnetometry, and infrared spectroscopy. The polymer had an average diameter of 250 ± 2.56 nm and exhibited good stability and adsorption for template molecule, which is enriched by hydrogen bonding interaction. Multiple factors for extraction, including loading, washing, elution solvents, and extraction time, were optimized. The limit of detection was 1.23 µg/g. The precision determined at various concentration of dihydroquercetin was less than 4% and the mean recovery was between 74.64 and 101.80%. It has therefore been shown that this protocol can be used as an alternative extraction to quantify dihydroquercetin in L. griffithiana and purify quercetin flavonoid from other complex matrices.

Endoscopic resection of giant gastrointestinal stromal tumor at the esophagogastric junction: a case report.

BACKGROUND: Gastrointestinal stromal tumors (GISTs) at the esophagogastric junction are rare and its treatment is complicated and challenging. Endoscopic resection has advantages with less complications compared to open and laparoscopic surgery. CASE PRESENTATION: We report a 33-year-old male patient who was admitted to our department complaining of abdominal fullness for 20 days. A huge submucosal tumor at the esophagogastric junction was found by upper gastrointestinal endoscopy. We successfully resected the lesion through endoscopic submucosal excavation without complications, which was pathologically confirmed to be a GIST. The patient was discharged 5 days after operation and has been doing well, and there was no recurrence 8 months after the operation. CONCLUSION: ESE is possibly an effective and minimally invasive method of giant esophagogastric junction stromal tumor.

Extraction and identification of matrine-type alkaloids from Sophora moorcroftiana using double-templated molecularly imprinted polymers with HPLC-MS/MS.

Double-templated molecularly imprinted polymers with specific recognition of three matrine-type alkaloids were prepared using matrine and oxymatrine as the template molecules. An approach based on double-templated molecularly imprinted solid-phase extraction coupled with high-performance liquid chromatography and tandem mass spectrometry was then developed to extract and purify matrine, oxymatrine, and sophocarpine from Sophora moorcroftiana in the Tibetan plateau herbs. The polymers were characterized by Fourier-transform infrared spectroscopy and scanning electron microscopy. Their adsorption characteristics were evaluated using adsorption kinetics, isotherms, selectivity, and recycling experiments. This polymer exhibited excellent molecular recognition ability and good selectivity. The obtained polymers as adsorbent was further used for the determination of three matrine-type alkaloids coupled to high-performance liquid chromatography with tandem mass spectrometry, the recoveries of three matrines spiked at three concentration levels in samples were 73.25-98.42% (n = 5) with a relative standard deviation less than 6.82%. The limits of detection for the method were 9.23-15.42 µg/kg (S/N = 3). This proposed method was assessed to be an effective method for simultaneous extraction, isolation, and identification of matrine, oxymatrine, and sophocarpine from Sophora moorcroftiana.

[Helminth Infections in Goats in Nimu County of Tibet].

Postmortem examinations were made in 99 goats in Nimu County of Tibe, and parasites were collected and identified based on morphology. The collected parasites were categorized, and infection status was analyzed. The helminth infection rate was 100% among the goats, and all showed a pattern of mixed infection. The identified parasites belonged to 21 species, 15 genera, and 9 families. The Trichuris genus（36.4%） was the most prevailing among nematodes in the gastrointestinal tract; Paramphistomum cervi（60.6%） and Paramphistomum gotoi（60.6%） were predominant among trematodes detected; Cysticercus tenuicollis（52.5%） was the predominant cestode detected; and Orientobilharzia turkestanicum was the major parasite detected in the portal vein (69.7%).

[Effect of Fuzhenghuayu compound on hepatocyte expression of Nrf2 in a mouse model of hepatic fibrosis].

OBJECTIVE: To investigate the effect of Fuzhenghuayu compound (FZHc) on expression of nuclear factor E2-related factor 2 (Nrf2) in hepatocytes under conditions of hepatic fibrosis using a mouse model. METHODS: Mice were randomly assigned to a control group and a hepatic fibrosis model group. The control group was further divided into three subgroups for use as normal controls (A1), mineral oil-treated controls (A2), and FZHc-treated controls (A3); the hepatic fibrosis model group was administered carbon tetrachloride (CC14 dissolved in mineral oil and injected intraperitoneally) and further divided into four subgroups for use as 6-weeks models (B1), 10-weeks models (B2), low-dose (L)-FZHc models (C1), and high-dose (H)-FZHc models (C2). The FZHc (capsule powder diluted with double-distilled water to 0.1 g/mL) was administered via gastric perfusion to groups A3, C1, and C2 starting at week 7 of the experiment. At the end of week 6 and 10, hepatic specimens were collected and evaluated for degree of hepatic fibrosis and inflammation using routine haematoxylin-eosin staining and Masson staining. Immunohistochemical analysis was performed to measure the hepatocyte expression of Nrf2, NAD(P)H quinine oxidoreductase 1 (Nqol), a-smooth muscle actin (a-SMA) and fibronectin (FN). Real-time fluorescence quantitative PCR was used to measure Nrf2 mRNA expression. Western blotting was used to detect Nrf2 and Nqol total protein expression and Nrf2 nuclear translocation. F test, LSD test and ridit test were used for statistical analyses. RESULTS: Compared with the B2 group (ridit value: 0.09), the model groups treated with FZHc showed significantly lower degrees of hepatic inflammation and fibrosis for both the low (C1 group, ridit value: 0.32) and high doses (C2 group, ridit value: 0.40) (F =82.927, P less than 0.05). In addition, compared with the B2 group, the model groups treated with FZHc showed significantly decreased expression of a-SMA and FN proteins, with a dose-dependent trend (by immunohistochemistry: C 1 group at the end of 10 weeks, F =77.421, 118.262, P less than 0.05; C2 group, P =0.002, 0.013) and significantly increased expression of Nrf2 and Nqol proteins (by immunohistochemistry:C1 and C2 groups at the end of 10 weeks, F =182.537, 75.615, P less than 0.05 and by westen blotting: F =45.664, 127.673, P less than 0.05), which also showed a dose-dependent trend (C2 group, P =0.000, 0.014; 0.005, 0.014). Western blotting also indicated that the amount of nuclear transported Nrf2 was higher in the C1 and C2 groups at the end of 10 weeks (vs. B2 group, F =94.787, P less than 0.05), and the amount of nuclear transported Nrf2 was significantly higher in the C2 group (vs. C1 group, P =0.044). Nrf2 mRNA expression was significantly higher in the C1 group than in the B2 group (F =3230.105, P less than 0.05), and the C2 group had more substantially increased expression (P =0.001); there was no statistical difference found between groups B1 and B2 (P =0.094). CONCLUSION: Fuzhenghuayu compound increased the expression of Nrf2 mRNA and protein under conditions of hepatic fibrosis in mice and stimulated Nrf2 nuclear transport, as well as increased expression of the Nrf2 target gene Nqol that is known to suppress activation of hepatic stellate cells and decrease the deposition of FN. Therefore, Fuzhenghuayu compound may ameliorate hepatocyte injury in hepatic fibrosis in mice by exerting an antihepatic fibrosis effect.

Adult diffuse hepatic hemangiomatosis lesion occupying the entire abdominal and pelvic cavities: a case report.

Introduction: Adult diffuse hepatic hemangiomatosis (DHH) is an extremely rare disease. Consequently, its characteristics are poorly understood. Herein, we report a case of adult DHH involving both liver lobes but without extrahepatic involvement. To the best of our knowledge, this the largest reported adult DHH to date. Case presentation: A 51-year-old man was admitted due to abdominal distension and dyspnea. Physical examination revealed marked liver enlargement. Color Doppler, plain and contrast-enhanced computed tomography, and contrast-enhanced magnetic resonance imaging revealed a hepatic lesion sized 35.1 × 32.1 × 14.1 cm occupying nearly the entire abdominal and pelvic cavities. Diagnosis was established by liver puncture biopsy. The patient exhibited clinical signs of portal hypertension and hypersplenism, but remains free of serious DHH-related complications. He is followed up regularly, with proactive evaluation for future liver transplantation. Conclusion: This case will contribute to the current knowledge on the clinical and imaging features of this rare entity.

Detection of early hidden carcinoma of cardia by reversely entering the esophagus with magnetically controlled capsule endoscopy: A case report.

Magnetically controlled capsule endoscopy (MCE) is a non-invasive method for gastropathy examination. However, due to the influence of gravity and lumen structure, the traditional capsule endoscopy rapidly passes through the cardia, leading to insufficient observation of the cardia mucosa. Case Summary. The patient, a 53-year-old male, had a history of subarachnoid hemorrhage for 5 years, and it has been 5 years since the aneurysm embolization.Computed Tomography Angiography (CTA) indicated the presence of an anterior cruciate aneurysm. Given the risks associated with traditional intubated gastroscopy, magnetic controlled capsule gastroscopy was chosen for gastric examination. Following the standard operating procedure, routine magnetic controlled capsule endoscopy was performed, and no lesions were detected.We combined magnetic force and patient posture adjustment to guide the capsule to pass through the cardia slowly and return to the esophagus, successfully detecting a concealed cardia lesion.Afterwards, the lesions of the cardia were treated with a magnifying gastroscope and endoscopic submucosal dissection (ESD).Pathological findings showed that adenocarcinoma was confined to the mucosa membrane, and in the postoperative pathological study, no tumor remnants or metastasis were discovered. This paper reports a case of a patient undergoing a physical examination, but no lesion was found during a routine examination using the magnetically controlled capsule gastroscope. However, we discovered a case of hidden early cardia cancer after guiding the capsule gastroscope back into the esophagus under magnetic control.

Network pharmacology analysis of the regulatory effects and mechanisms of ALAE on sow reproduction in vivo and in vitro.

ETHNOPHARMACOLOGICAL RELEVANCE: Reproductive ability of sows is a primary element influencing the development of pig farming. Herbal extracts of Angelica sinensis (Oliv.) Diels, Astragalus mongholicus Bunge, Eucommia ulmoides Oliv., and Polypodium glycyrrhiza D.C.Eaton showed effects on improvement of reproduction in sows. AIMS OF THE STUDY: To investigate the mechanism of the treatment effects by a compound of these four Chinese herbs in a 1:1:1:1 ratio (ALAE) on endometriosis, endometritis, uterine adhesion, intrauterine growth retardation, pre-eclampsia, and its enhancement of reproductive efficiency in sows. MATERIALS AND METHODS: Active components of ALAE were identified by using ultra-performance liquid chromatography-mass spectrometry analysis and network pharmacology. Then we used the results to construct a visualization network. Key targets and pathways of ALAE involved in sow reproduction improvement were validated in sow animals and porcine endometrial epithelial cells (PEECs). RESULTS: A total of 62 active compounds were found in ALAE (41 in Polypodium glycyrrhiza D.C.Eaton, 5 in Astragalus mongholicus Bunge, 11 in Eucommia ulmoides Oliv., 5 in Angelica sinensis (Oliv.) Diels) with 563 disease-related targets (e.g. caspase-3, EGFR, IL-6) involved in EGFR tyrosine kinase inhibitor resistance, PI3K-AKT, and other signaling pathways. Molecular docking results indicated GC41 (glabridin), GC18 (medicarpin), EGFR and CCND1 are possible key components and target proteins related to reproductive improvement in sows. In PEECs, EGFR expression decreased at the mRNA and protein levels by three doses (160, 320, and 640 µg/mL) of ALAE. The phosphorylation of downstream pathway PI3K-AKT1 was enhanced. The expression of inflammatory factors (IL-6, IL-1ß), ESR1 and caspase-3 decreased through multiple pathways. Additionally, the expression levels of an anti-inflammatory factor (IL-10), angiogenesis-related factors (MMP9, PIGF, PPARγ, IgG), and placental junction-related factors (CTNNB1, occludin, and claudin1) increased. Furthermore, the total born number of piglets, the number of live and healthy litters were significantly increased. The number of stillbirths decreased by ALAE treatment in sow animals. CONCLUSIONS: Dministration of ALAE significantly increased the total number of piglets born, the numbers of live and healthy litters and decreased the number of stillbirths through improving placental structure, attenuating inflammatory response, modulating placental angiogenesis and growth factor receptors in sows. The improvement of reproductive ability may be related to activation of the EGFR-PI3K-AKT1 pathway in PEECs. Moreover, ALAE maybe involved in modulation of estrogen receptors, apoptotic factors, and cell cycle proteins.

Siraitia grosvenorii Extract Protects Lipopolysaccharide-Induced Intestinal Inflammation in Mice via Promoting M2 Macrophage Polarization.

Siraitia grosvenorii has anti-inflammatory, antioxidant, and immune-regulating effects, while macrophages play an important role in reducing inflammation. However, it is still unclear whether Siraitia grosvenorii extract (SGE) is effective in reducing inflammation by regulating macrophages. This study investigated the regulatory effect of SGE on macrophage polarization in a lipopolysaccharide (LPS)-induced intestinal inflammation model after establishing the model in vitro and in vivo. The results from the in vivo model showed that, compared with the LPS group, SGE significantly improved ileal morphology, restored the ileal mucosal barrier, and reduced intestinal and systemic inflammation by increasing CD206 and reducing iNOS proteins. In the in vitro model, compared with the LPS group, SGE significantly reduced the expression of iNOS protein and cytokines (TNF-α, IL-1ß, and IFN-γ) while significantly increasing the protein expression of CD206 in RAW264.7 cells. In conclusion, SGE can alleviate intestinal inflammation, protect the mucus barrier, and block the systemic immunosuppressive response by increasing M2 macrophages.

Preliminary Proteomic Study of the Porcine Pituitary Gland under Heat Stress.

Although numerous studies have shown that the hypothalamic-pituitary-adrenal axis plays a vital role in the response to environmental stress by mediating the production of a series of hormones, the mechanism underlying these effects has not been elucidated. This study used proteomics techniques to investigate the differentially expressed proteins (DEPs) in the pituitary glands of pigs and to elucidate the potential changes in the immune-neuroendocrine system under heat stress (HS). In total, 2517 peptides corresponding to 205 proteins were detected. A comparison of the expression patterns between HSs and healthy controls revealed 56 DEPs, of which 31 were upregulated and 25 were downregulated. Ingenuity pathway analysis (IPA) was used to reveal the subcellular characteristics, functional pathways, regulatory networks, and upstream regulators of the identified proteins. The results showed that these differentially expressed proteins were involved in intercellular communication, interactions, apoptosis, nervous system development, functions, abnormalities and other functions, and in the regulatory network. Moreover, the upstream regulators of the differentially expressed proteins were mainly transcriptional regulators, hormones, and cytokines. Thus, the functional network and pathway analyses could provide insights into the complexity and dynamics of HS-host interactions and may accelerate our understanding of the mechanisms underlying HS.

Protective Effects of Carbonated Chitosan Montmorillonite on Vomitoxin-Induced Intestinal Inflammation.

Exposure to vomitoxin (DON) can negatively impact the intestinal health of livestock and poultry, leading to compromised nutrient absorption and utilization, resulting in slowed growth and reduced production efficiency. In this study, we synthesized carbonated chitosan montmorillonite intercalation complexes (CCM) through solution precipitation. The successful formation of intercalation complexes was confirmed by examining functional groups and surface features using infrared spectroscopy and scanning electron microscopy. To assess the impact of CCM on DON-infected mice, we established an experimental mouse model of jejunal inflammation induced by DON infection. We analyzed the effects of CCM on blood biochemical and conventional indices, jejunal inflammatory factors, pathological changes, and the expression of proteins in the MAPK pathways in DON-infected mice. Our results indicate that CCM effectively mitigates the adverse effects of DON on growth performance, jejunal injury, and the inflammatory response in mice. CCM supplementation alleviated the negative effects of DON infection on growth performance and reduced intestinal inflammation in mice. Moreover, CCM supplementation successfully inhibited the activation of the mitogen-activated protein kinase (MAPK) signaling pathway induced by DON. These findings suggest that the mitigating effect of CCM on DON-induced inflammatory injury in the murine jejunum is closely linked to the regulation of the MAPK signaling pathway.

Baicalin methyl ester prevents the LPS - induced mice intestinal barrier damage in vivo and in vitro via P65/TNF-α/MLCK/ZO-1 signal pathway.

The effect of baicalin methyl ester (BME) on the regulation of mice intestinal barrier in the inflammatory response was studied in vivo and in vitro. Thirty six C57/BL mice were randomly divided into six groups (n = 6): control group; LPS group (LPS 3.5 mg/kg given intraperitoneal [ip] on day 7 of the study only), PBS group, and three BME groups (low: 50 mg/kg; medium: 100 mg/kg; high: 200 mg/kg) orally dosed with BME for 7d and LPS ip on day 7. All mice were sacrificed on day 8, and jejunum tissue collected for histopathology (H&E and PAS staining), protein expression of pro-inflammatory factors (TNF-α, IL-6, IL-8, IFN-γ) by ELISA, and intestinal tight junction proteins (ZO-1, occludin, claudin-1 and claudin-4) by Western Blot. Compared with the control group, LPS significantly increased the serum cytokines DAO (p < 0.01) and DLA (p < 0.01), upregulated the expression of pro-inflammatory factors, MLCK proteins (p <0.05) and increased the MLCK/ZO-1ratio (p <0.001). LPS also decreased the expression of claudin-4 (p < 0.01) in the jejunum and induced an inflammatory response damaging the jejunal mucosal barrier. Pretreatment with BME (100-200 mg/kg) significantly decreased the cytokines DAO (p < 0.05) and DLA (p < 0.01) in the serum, pro-inflammatory factors in the jejunum, significantly down-regulated the expression of MLCK (p <0.05) and the ratio of MLCK/ZO-1(p <0.001) but upregulated the expressions of ZO-1(p < 0.01), occludin (p < 0.05), claudin-1(p < 0.05) and claudin-4 (p < 0.05), and thereby restored the intestinal tissue structure, suggestive of alleviation of LPS-induced intestinal inflammation by BME. In vitro, MODE-K cells (derived from mice intestinal epithelium) were exposed to BME at 0 (control group-No LPS), 10, 20 and 40 µM BME for 24 h prior to LPS addition at 50 µg/mL for 2 h. LPS significantly increased the expression of pro-inflammatory factors, MLCK (p < 0.01) and the ratio of MLCK/ZO-1(p <0.001), decreased the expressions of ZO-1 (p < 0.05), occludin (p < 0.01), claudin-1 (p < 0.01) and claudin-4 (p < 0.01) in MODE-K cells compared with the control group. Compared with the LPS group, BME (10 - 40 µM) significantly decreased the expression of pro-inflammatory factors, MLCK (p < 0.05) and the ratio of MLCK/ZO-1(p <0.01) but increased the expressions of ZO-1(p < 0.01), occludin (p < 0.05) and claudin-4(p < 0.01) indicating an up-regulation of the expression of tight junction proteins by BME. On addition of extrinsic TNF-α plus LPS, the TNF- α level increased (p < 0.001) in MODE-K cells and the protein expression of MLCK (p < 0.01) was markedly up-regulated. Molecular docking predicted BME interacted with P65 by forming hydrogen bonds. IP-WB further confirmed that BME was directly bound to P65 protein in MODE-K cells. In conclusion, BME was able to restore the intestinal barrier through the P65 / TNF-α / MLCK / ZO-1 signaling pathway.

Chitosan-gentamicin conjugate attenuates heat stress-induced intestinal barrier injury via the TLR4/STAT6/MYLK signaling pathway: In vitro and in vivo studies.

Heat stress (HS) has a negative impact on animal health. A modified chitosan-gentamicin conjugate (CS-GT) was prepared to investigate its potential protective effects and mechanism of action on heat stress-induced intestinal mucosa injury in IPEC-J2 cells and mouse 3D intestinal organs in a mouse model. CS-GT significantly (P < 0.01) reversed the decline in transmembrane resistance and increased the FITC-dextran permeability of the IPEC-J2 monolayer fusion epithelium caused by heat stress. Heat stress decreased the expression of the tight binding proteins occludin, claudin1, and claudin2. However, pretreatment with CS-GT significantly increased (P < 0.01) the expression of these tight binding proteins. Mechanistically, CS-GT inhibited the activation of the TLR4/STAT6/MYLK signaling pathway induced by heat stress. Molecular docking showed that CS-GT can bind effectively with TLR4. In conclusion, CS-GT alleviates heat stress-induced intestinal mucosal damage both in vitro and in vivo. This effect is mediated, at least partly, by the inhibition of the TLR4/STAT6/MYLK signaling pathway and upregulation of tight junction proteins. These findings suggest that CS-GT may have therapeutic potential in the prevention and treatment of heat stress-related intestinal injury.