Results Of Immunohistochemical Staining Research Articles

Programmed cell death factor 4-mediated hippocampal synaptic plasticity is involved in early life stress and susceptibility to depression

Early life stress (ELS) increases the risk of depression later in life. Programmed cell death factor 4 (PDCD4), an apoptosis-related molecule, extensively participates in tumorigenesis and inflammatory diseases. However, its involvement in a person’s susceptibility to ELS-related depression is unknown. To examine the effects and underlying mechanisms of PDCD4 on ELS vulnerability, we used a “two-hit” stress mouse model: an intraperitoneal injection of lipopolysaccharide (LPS) into neonatal mice was performed on postnatal days 7–9 (P7-P9) and inescapable foot shock (IFS) administration in adolescent was used as a later-life challenge. Our study shows that compared with mice that were only exposed to the LPS or IFS, the “two-hit” stress mice developed more severe depression/anxiety-like behaviors and social disability. We detected the levels of PDCD4 in the hippocampus of adolescent mice and found that they were significantly increased in “two-hit” stress mice. The results of immunohistochemical staining and Sholl analysis showed that the number of microglia in the hippocampus of “two-hit” stress mice significantly increased, with morphological changes, shortened branches, and decreased numbers. However, knocking down PDCD4 can prevent the number and morphological changes of microglia induced by ELS. In addition, we confirmed through the Golgi staining and immunohistochemical staining results that knocking down PDCD4 can ameliorate ELS-induced synaptic plasticity damage. Mechanically, the knockdown of PDCD4 exerts neuroprotective effects, possibly via the mediation of BDNF/AKT/CREB signaling. Combined, these results suggest that PDCD4 may play an important role in the ELS-induced susceptibility to depression and, thus, may become a therapeutic target for depressive disorders.

The utilization of chitosan/bletilla striata hydrogels to elevate anti-adhesion, anti-inflammatory and pro-angiogenesis properties of polypropylene mesh in abdominal wall repair

Abstract Polypropylene (PP) mesh is commonly used in abdominal wall repair due to its ability to reduce the risk of organ damage, infections, and other complications. However, the PP mesh often leads to adhesion formation and does not promote functional tissue repair. In this study, we synthesized one kind of aldehyde Bletilla striata polysaccharide (BSPA) modified chitosan (CS) hydrogel based on Schiff base reaction. The hydrogel exhibited a porous network structure, a highly hydrophilic surface, and good biocompatibility. We wrapped the PP mesh inside the hydrogel and evaluated the performance of the resulting composites in a bilateral 1 × 1.5 cm abdominal wall defect model in rats. The results of gross observation, histological staining, and immunohistochemical staining demonstrated the positive impact of the CS hydrogel on anti-adhesion and wound healing effects. Notably, the addition of BSPA to the CS hydrogel further improved the performance of the composites in vivo, promoting wound healing by enhancing collagen deposition and capillary rearrangement. This study suggested that the BSPA-modified CS hydrogel significantly promoted the anti-adhesion, anti-inflammatory and pro-angiogenesis properties of PP meshes during the healing process. Overall, this work offers a novel approach to the design of abdominal wall repair patches.

Bioinformatics-based analysis of the effect of general Transcription Factor IIH on prognosis of Prostate cancer

Objective: To investigate the genetic and expression characteristics of transcription factor IIH (TFIIH) in pre-initiationcomplex in prostate cancer (PCa) and its relationship with prostate cancer progression. Methods: Analyzing the expression characteristics and clinical signification of TFIIH subunits about 495 cases of PCa and 52 cases of adjacent cancer in The Cancer Genome Atlas-Prostate adenocarcinoma (TCGA-PRAD) database. PCa microarray chip was used to verify the correlation between the key factor General Transcription Factor IIH Subunit 4 (GTF2H4) in TFIIH and clinical features. Results: The 495 patients with PCa were (61.01±6.82) years old.The mRNA expression of ERCC3、GTF2H4 and MNAT1 were high in PCa tissues with GS≥8(P<0.05). The expression of GTF2H4 and MNAT1 were relevant to the pathological stages(P<0.05). High expression of GTF2H4 has higher biochemical recurrence (BCR) rate in PCa patients(HR=2.47, 95%CI:1.62-3.77, P<0.001), which has better predictive effect of BCR in PCa patients(The 3rd, 5th, and 7th year AUC all>0.7) than other subunits, and it has been verified in four additional databases. Single-factor Cox regression analysis showed that GTF2H4 were risk factors for BCR (HR=2.470, 95%CI:1.620-3.767, P<0.001) and GTF2H5 were protective factors(HR=0.506,95%CI: 0.336-0.762, P=0.001). The results of immunohistochemical staining showed that the protein expression of GTF2H4 was correlated with the clinical features of PCa patients.The differences of the above results were statistically significant. Conclusion: GTF2H4, the key factor of TFIIH, is highly expressed in PCa and indicates a poor prognosis.

Transcriptome sequencing reveals novel biomarkers and immune cell infiltration in esophageal tumorigenesis.

Esophageal squamous cell carcinoma (ESCC) is one of the most common malignancies worldwide, and its development comprises a multistep process from intraepithelial neoplasia (IN) to carcinoma (CA). However, the critical regulators and underlying molecular mechanisms remain largely unknown. To explore the genes and infiltrating immune cells in the microenvironment that are associated with the multistage progression of ESCC to facilitate diagnosis and early intervention. A mouse model mimicking the multistage development of ESCC was established by providing warter containing 4-nitroquinoline 1-oxide (4NQO) to C57BL/6 mice. Moreover, we established a control group without 4NQO treatment of mice. Then, transcriptome sequencing was performed for esophageal tissues from patients with different pathological statuses, including low-grade IN (LGIN), high-grade IN (HGIN), and CA, and controlled normal tissue (NOR) samples. Differentially expressed genes (DEGs) were identified in the LGIN, HGIN, and CA groups, and the biological functions of the DEGs were analyzed via Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses. The CIBERSORT algorithm was used to detect the pattern of immune cell infiltration. Immunohistochemistry (IHC) was also conducted to validate our results. Finally, the Luminex multiplex cytokine analysis was utilized to measure the serum cytokine levels in the mice. Compared with those in the NOR group, a total of 681541, and 840 DEGs were obtained in the LGIN, HGIN, and CA groups, respectively. Using the intersection of the three sets of DEGs, we identified 86 genes as key genes involved in the development of ESCC. Enrichment analysis revealed that these genes were enriched mainly in the keratinization, epidermal cell differentiation, and interleukin (IL)-17 signaling pathways. CIBERSORT analysis revealed that, compared with those in the NOR group, M0 and M1 macrophages in the 4NQO group showed stronger infiltration, which was validated by IHC. Serum cytokine analysis revealed that, compared with those in the NOR group, IL-1β and IL-6 were upregulated, while IL-10 was downregulated in the LGIN, HGIN, and CA groups. Moreover, the expression of the representative key genes, such as S100a8 and Krt6b, was verified in external human samples, and the results of immunohistochemical staining were consistent with the findings in mice. We identified a set of key genes represented by S100a8 and Krt6b and investigated their potential biological functions. In addition, we found that macrophage infiltration and abnormal alterations in the levels of inflammation-associated cytokines, such as IL-1β, IL-6, and IL-10, in the peripheral blood may be closely associated with the development of ESCC.

A case of pancreatic PEComa with prominent inflammatory cell infiltration: the inflammatory subtype is a distinct histologic group of PEComa

BackgroundPEComa is a mesenchymal tumor that can occur in various organs including the uterus and soft tissues. PEComas are composed of perivascular epithelioid cells, and angiomyolipoma (AML), clear cell sugar tumor (CCST), and lymphangiomyomatosis (LAM) are considered lesions of the same lineage as tumors of the PEComa family. Histologically, a common PEComa shows solid or sheet-like proliferation of epithelioid cells. This is accompanied by an increase in the number of dilated blood vessels. Here, we report a case of pancreatic PEComa with marked inflammatory cell infiltration.Case presentationA 74-year-old male patient underwent an appendectomy for acute appendicitis. Postoperative computed tomography and magnetic resonance imaging revealed a 30 × 25 mm non-contrast-enhanced circular lesion in the tail of the pancreas. The imaging findings were consistent with a malignant tumor, and distal pancreatectomy was performed. Histologically, most area of the lesion was infiltrated with inflammatory cells. A few epithelioid cells with large, round nuclei, distinct nucleoli, and eosinophilic granular cytoplasm were observed. Spindle-shaped tumor cells were observed. Delicate and dilated blood vessels were observed around the tumor cells. Immunohistochemically, the atypical cells were positive for αSMA, Melan A, HMB-45, and TFE3. The cytological characteristics of the tumor cells and the results of immunohistochemical staining led to a diagnosis of pancreatic PEComa.ConclusionsA histological variant known as the inflammatory subtype has been defined for hepatic AML. A small number of tumor cells present with marked inflammatory cell infiltration, accounting for more than half of the lesions, and an inflammatory myofibroblastic tumor-like appearance. To our knowledge, this is the first report of pancreatic PEComa with severe inflammation. PEComa is also a generic term for tumors derived from perivascular epithelioid cells, such as AML, CCST, and LAM. Thus, this case is considered an inflammatory subtype of PEComa. It has a distinctive morphology that is not typical of PEComa. This histological phenotype should be widely recognized.

Clinicopathological and genetic characteristics of common lymphatic malformations in superficial soft tissues

Objective: To investigate the clinicopathological features, classification, and genetic characteristics of common lymphatic malformation (CLM) in superficial soft tissue. Methods: A retrospective study of 110 patients with the diagnosis of CLM at the Henan Province People's Hospital, China from August 2019 to August 2022 was performed. The clinicopathological features, relevant immunohistochemical (IHC) staining results, and fluorescence quantitative PCR of PIK3CA mutation were analyzed, and patients were followed up. Results: Among the 110 CLM patients, there were 53 males and 57 females; 65 cases (65/110, 59.1%) were first detected when the patients were≤2 years old. The most common location was the head and neck in 41 cases (41/110, 37.3%). Clinically, 102 cases (102/110, 92.7%) were solitary, 83 cases (83/110, 75.5%) were skin-colored, 69 cases (69/110, 62.7%) had indistinct borders, and 10 cases (10/110, 9.1%) had diffuse and severe macroscopic manifestations. There were 52 macrocystic type (52/110, 47.3%), 23 microcystic type (23/110, 20.9%), and 35 combined type (35/110, 31.8%). The macrocystic CLM presented as soft, translucent masses with large cystic cavities on the cut surface, and histologically they were composed of large, irregularly dilated channels that were thicker with irregular smooth muscle and lymphocytic infiltration. Microcystic CLM showed wartlike projections or translucent blisters on the skin, with small honeycomb structures on the cut surface, and histologically consisted of round or angular dilated small lymphatic vessels with little or no smooth muscle. The combined CLM had both macrocystic and microcystic morphologies. IHC staining showed that the lymphatic endothelial cells were positive for LYVE-1, D2-40, PROX1, CD31, and VEGFR3 but negative for CD34; in the macrocystic and combined CLM vessel walls were positive for SMA. Eight of 13 CLM had PIK3CA mutation. All patients were followed up, and 24 (24/110, 21.8%) had relapses, which more frequently occurred in combined type, followed by microcystic type. Conclusions: CLM is a congenital vascular malformation composed of dilated, abnormal lymphatic channels, with PIK3CA mutation. There are significant differences in clinicopathological characteristics among the different types. Since microcystic and combined CLM are prone to recurrence, accurate pathological subtyping is necessary to guide treatment and to predict prognosis.

Sarcomatoid renal pelvis carcinoma: Experience of treatment at a single-institution

ObjectivesSarcomatoid renal pelvis carcinoma (SRPC) is a rare variant of RPC. We aimed to summarize the clinicopathological features and prognostic factors of SRPC. MethodsIn this retrospective study, we collected data from 24 patients with SRPC who were treated at the Department of Urology, Affiliated Hospital of Qingdao University between 2008 and 2021. The clinicopathological features of the patients were obtained from their medical records to evaluate the diagnosis, prognostic factors, and response to systemic therapy. ResultsImmunohistochemical staining revealed that cytokeratin was expressed in 19 patients with SRPC, while vimentin was expressed in all patients. Computer tomography showed these tumors as low-density (n = 12) or mixed-density masses, with or without necrotic areas (n = 12). All patients showed different degrees of enhancement on computed tomography. Lymph node metastasis was present in 6 patients and distant metastasis in 5. The median survival of all patients was 28 months. Patients without metastasis had a median survival of 46 months compared with 18 months in those with metastasis (P < 0.05). Necrosis had no significant influence on prognosis (P > 0.05). The median survival of patients with and without hydronephrosis was 18 and 104 months (P < 0.05). Among patients without metastasis, those without hydronephrosis survived longer than those with hydronephrosis (104 vs 18 months, P < 0.05), and necrosis had no effect on prognosis. In patients with metastasis, necrosis and hydronephrosis had no effect on prognosis (P > 0.05). ConclusionThe prognosis of SRPC is poor, and the clinical stage, particularly the presence of distant metastasis, has a significant impact on prognosis.

Notch signaling genes and CD8+ T-cell dynamics: Their contribution to immune-checkpoint inhibitor therapy in oral squamous cell carcinoma: A retrospective study.

Aberrant Notch signaling pathway has been related with the tumorigenesis in head and neck region, involving oral cavity. Here, we report the correlation between mutations in the Notch signaling pathway and CD8+ T-cell infiltration via PD-L1, which lead to enhanced antitumor immunity and may target for immune-checkpoint inhibitors (ICIs) therapy. This retrospective study analyzed the results of immunohistochemical staining for PD-L1 and CD8+ T-cell infiltration in 10 patients and whole-exome sequencing (WES) was conducted on five of these patients to identify frequently mutated genes. Four of 10 patients were positive for PD-L1 and CD8+ T. By analyzing WES in three of these four patients, we notably identified the mutations of NOTCH1, FBXW7, and noncoding RNA intronic mutation in NOTCH2NLR in two of these three patients. This study may enable better selection of ICI therapy with CD8+ T-cell infiltration via PD-L1 expression for oral squamous cell carcinoma patients with mutations in Notch signaling pathway.

Orthodontic tension promotes cementoblast mineralization by regulating autophagy

Background/purposeExternal root resorption is a main side effect of orthodontic treatment and is more likely to occur on the pression side than the tension side. To explore the potential protective mechanisms on the tension side, this study investigated the influence of mechanical tension on cementoblast mineralization and elucidated the role of autophagy in mediating this process. Materials and methodsMechanical tension was applied to cementoblasts using iStrain. The expression of mineralization-related and autophagy-related markers was detected by qRT-PCR, Western blot analysis, and immunofluorescence staining. RNA sequencing identified key regulators. Immunohistochemical staining assessed related markers expression in in vivo experiments. ResultsApplying tension to cementoblasts increased mineralization-related gene expression in a force-dependent and time-dependent manner. The immunohistochemical staining result of in vivo experiments supported these findings, demonstrating elevated expression of mineralization markers under tension. Mechanical tension also enhanced autophagic activity in cementoblasts, which was demonstrated by the results of qRT-PCR, Western blot analysis, immunofluorescence staining, and in vivo experiments. Suppression of autophagy with chloroquine attenuated the mineralization of cementoblasts induced by tension stimulus. RNA-seq identified Postn as a key regulator, and the knockdown of Postn impaired the mechanical tension-promoted mineralization of cementoblasts. ConclusionThis study proposed the tension-induced promotion in mineralization of cementoblasts and emphasized the mediating role of autophagy in this process. Postn, a mediator connecting autophagy and mineralization, was identified as a key regulator. These discoveries helped elucidate orthodontic-related microprocesses on tooth roots and offer potential targets for therapeutic interventions to prevent and restore external root resorption clinically.

The Effect of Necrostatin -1 and Enoxaparin Molecules on Random Pattern Flap Viability

Distal flap necrosis is seen more often in random pattern flaps and is an important complication that shortens the flap length. There has been much research many drugs and molecules in an effort to prevent this complication. The aim of this study was to investigate the efficacy of necrostatin-1 and enoxaparin molecules in preventing distal flap necrosis and increasing flap viability in a random pattern flap model created in rats. A total of 32 Wistar albino female rats, each weighing 300-350 gr were separated into 4 groups. All the animals underwent an operation to create a 3×9 cm caudal-based Mcfarlane flap. The treatments defined for each group were applied. Full layer tisssue samples 1×1 cm2 were taken from all the flaps and stored until histopathological and immunohistochemical examination, the parameters of inflammation, capillary proliferation, necrosis, fibroblast proliferation and fibrosis were compared histopathologically. In the necrostatin-1 group, the inflammation, necrosis and fibrosis scores were observed to be lower and the capillary proliferation and fibroblast proliferation scores were higher. In the enoxaparin group, the fibroblast proliferation and capillary proliferation scores were higher. The receptor interacting protein kinase-1 immunohistochemical staining results showed statistically significantly less staining in the necrostatin-1 group compared to the other groups. The results of this study suggest that necrostatin molecule has important therapeutic potential in increasing flap viability in the random pattern flap model, considering the percentage of flap necrosis, and the immunohistochemical and histopathological data. The flap necrosis percentage and histochemical parameters of the enoxaparin molecule demonstrate that the effects on flap viability are limited.

MUC1 promotes RIF by regulating macrophage ROS-SHP2 signaling pathway to up-regulate inflammatory response and inhibit angiogenesis.

To explore the effect of MUC1 on recurrent implantation failure (RIF) and its molecular mechanism. Bioinformation analysis was used to find possible molecular mechanisms of specific genes in the pathogenesis of RIF. The number of M1 and M2 macrophages was measured by flow cytometry. Immunohistochemical staining and western blotting were used to detect the expression of related proteins. Angiogenesis capacity was measured by cell tube-formation assay. Bioinformatics analysis results suggest that MUC1 may play an important role in RIF. The results of flow cytometry showed that compared with NC group, M1 macrophages increased significantly and M2 macrophages decreased significantly in MUC1 OE group. The results of immunohistochemical staining showed that MUC1 could inhibit the expression of VEGF. Western blotting results showed that MUC1 could significantly increase the expression of P22, P47, gp91, p-TBK1, IFNγ and IL-1β, and decrease the expression of p-SHP2, p-PI3K, p-mTOR, HIF1α and VEGF. After the addition of ROS inhibitor and PI3K inhibitor, the effect of MUC1 on the above proteins was eliminated. The results of tube formation experiments showed that MUC1 could inhibit vascular formation. As a promising biomarker for the diagnosis of RIF, MUC1 can promote RIF by regulating macrophage ROS-SHP2 signaling pathway to up-regulate inflammatory response and inhibit angiogenesis.

Efficacy of immune checkpoint inhibitors in SMARCA4-deficient and TP53 mutant undifferentiated lung cancer.

The present study was conducted to characterize the clinicopathologic characteristics, immunohistochemical staining results, and immune checkpoint inhibitors (ICIs) efficacy in patients with SMARCA4-deficient/TP53 mutant lung cancer. Patients diagnosed with advanced or metastatic undifferentiated lung cancer harboring SMARCA4-deficient and TP53 mutations, however, without targetable sensitive mutations were retrieved from the electronic medical record system. Descriptive statistics were used to describe the baseline characteristics and clinical features including age, gender, eastern cooperative oncology group performance status, disease stage, smoking status, chief complaint, site of the primary mass, tumor size, gross type, symptoms, local invasion, and metastatic sizes. Immunological markers and potential drive genes were detected by immunohistochemical staining and next generation sequencing. Efficacy and safety profile of ICIs in included patients was evaluated with progression-free survival and overall survival. Between January 2019 and September 2022, there were 4 patients included within the inclusion criteria in the present study. Biomarkers including CK, CK7, and integrase interactor 1 were detected positive, however, other immunological markers including CK20, CD56, P63, P40, NapsinA, TTF-1, CgA, Syn, BRG1, or PD-L1 were detected negative among them. Results of next generation sequencing panel were failed to discover any targetable sensitive mutations. A total of 4 mutation types of TP53, including p.C141Y, p.S240G, p.E339X (terminator acquired), and p.L130F detected for the patients, respectively. Microsatellite stability status, as well as low tumor mutation burden was identified among all the patients. Median progression-free survival for ICIs as first line treatment and median overall survival were 3.25 months (range from 1.3 to 6.8 months), and 6.0 months (range from 2.7 to 9.6 months), respectively. Our results indicated that advanced lung cancer patients harboring co-occurring SMARCA4-deficient/TP53 mutations might respond to ICIs treatment, though within negative programmed cell death-ligand 1 expression or low tumor mutation burden. However, hyperprogressive disease by ICIs may also happen for such patients. The mutation types of TP53 might play a role during the exposure of ICIs, however, need further identification in basic experiments.

Elucidating the mechanisms of formononetin in modulating atherosclerotic plaque formation in ApoE-/- mice

BackgroundAtherosclerosis(AS) poses a pressing challenge in contemporary medicine. Formononetin (FMN) plays a crucial role in its prevention and treatment. However, the detailed impact of FMN on the stability of atherosclerotic plaques and its underlying mechanisms remain to be elucidated.MethodsAn intervention consisting of FMN was given along with a high-fat food regimen in the ApoE-/- mouse model. The investigation included the evaluation of the degree of atherosclerotic lesion, the main components of the plaque, lipid profiles, particular markers indicating M1/M2 macrophage phenotypes, the quantities of factors related to inflammation, the infiltration of macrophages, and the identification of markers linked to the α7nAChR/JAK2/STAT3 axis effect molecules.ResultsThe evaluation of aortic morphology in ApoE-/-mice revealed that FMN significantly improved the plaque area, fibrous cap protrusion, lipid deposition, and structural alterations on the aortic surface, among other markers of atherosclerosis,and there is concentration dependence. Furthermore, the lipid content of mouse serum was assessed, and the results showed that the low-, medium-, and high-dosage FMN groups had significantly lower levels of LDL-C, ox-LDL, TC, and TG. The results of immunohistochemical staining indicated that the low-, medium-, and high-dose FMN therapy groups had enhanced CD206 expression and decreased expression of CD68 and iNOS. According to RT-qPCR data, FMN intervention has the potential to suppress the expression of iNOS, COX-2, miR-155-5p, IL-6, and IL-1β mRNA, while promoting the expression of IL-10, SHIP1, and Arg-1 mRNA levels. However, the degree of inhibition varied among dosage groups. Western blot investigation of JAK/STAT signaling pathway proteins and cholinergic α7nAChR protein showed that p-JAK2 and p-STAT3 protein expression was suppressed at all dosages, whereas α7nAChR protein expression was enhanced.ConclusionsAccording to the aforementioned findings, FMN can reduce inflammation and atherosclerosis by influencing macrophage polarization, blocking the JAK/STAT signaling pathway, and increasing α7nAChR expression.

The significance of an immunohistochemical marker-based panel in assisting the diagnosis of parathyroid carcinoma

Abstract Purpose Parathyroid carcinoma (PC) is an endocrine malignancy with a poor prognosis. However, the diagnosis of PC is still a difficult problem. A model with immunohistochemical (IHC) staining of 5 biomarkers has been reported from limited samples for the differential diagnosis of PC. In the present study, a series of IHC markers was applied in relatively large samples to optimize the diagnostic model for PC. Methods In this study, 44 patients with PC, 6 patients with atypical parathyroid tumors and 57 patients with parathyroid adenomas were included. IHC staining for parafibromin, Ki-67, galectin-3, protein-encoding gene product 9.5 (PGP9.5), E-cadherin, and enhancer of zeste homolog 2 (EZH2) was performed on formalin-fixed, paraffin-embedded tissue samples. The effects of clinical characteristics, surgical procedure, and IHC staining results of tumor tissues on the diagnosis and prognosis of PC were evaluated retrospectively. Results A logistic regression model with IHC results of parafibromin, Ki-67, and E-cadherin was created to differentiate PC with an area under the curve of 0.843. Cox proportional hazards analysis showed that negative parafibromin staining (hazard ratio: 3.26, 95% confidence interval: 1.28–8.34, P = 0.013) was related to the recurrence of PC. Conclusion An IHC panel of parafibromin, Ki-67 and E-cadherin may help to distinguish PC from parathyroid neoplasms. Among the 6 IHC markers and clinical features examined, the risk factor related to PC recurrence was parafibromin staining loss.

Effect of huankuile on colon injury in rats with ulcerative colitis by reducing TNF-α and MMP9

ObjectiveTo explore the mechanism of huankuile (HKL) in colon injury repair in rats with ulcerative colitis (UC).MethodsFifty SPF Wistar male rats were divided randomly into a normal group, a negative control group, an HKL intervention group (‘HKL group’) and a 5-aminosalicylic acid intervention group (‘5-ASA group’). After 14 days of intervention with corresponding drugs, pathological scores were obtained using the results of immunohistochemical staining; morphological changes were observed by hematoxylin–eosin staining, and the mRNA expression levels of tumour necrosis factor-α (TNF-α), matrix metalloproteinase 9 (MMP9) and interleukin-13 (IL-13) were detected by real-time quantitative PCR.ResultsAfter the successful construction of the rat model, it was compared with the rats in the normal group. In the negative group, it was found that the expression of TNF-α and MMP9 was significantly increased in the colonic mucosal epithelia of the rats, the pathological score was significantly increased (P < 0.05), and the mRNA expression levels of TNF-α, MMP9 and IL-13 were increased (P < 0.05). After treatment with HKL, the colonic morphology of the rats returned to normal, the expression of TNF-α and MMP9 in the colonic mucosal epithelium of the rats returned to normal, the pathological score grade was significantly reduced (P < 0.05), and the mRNA expression levels of TNF-α, MMP9 and IL-13 were reduced; these results were largely consistent with those of the normal group, with no statistically significant difference.ConclusionHKL effectively improved the general symptoms and tissue injury in UC rats, and the therapeutic effect was better than that of 5-ASA group. Ulcerative colitis in rats increased the expression of TNF-α, MMP9 and IL-13. HKL repaired UC-induced colonic injury in rats by decreasing the expression of TNF-α, MMP9 and IL-13.

MiR-22 inhibits myocardial fibrosis in rats with myocardial infarction by targeting PTEN/Akt/mTOR signaling pathway.

This study aimed to evaluate the effects of miR-22 on myocardial fibrosis in rats with myocardial infarction (MI) and to further explore the possible underlying mechanism. A total of 80 rats were randomly divided into Sham group, miR-22 overexpression group, MI group or MI + miR-22 overexpression group. Reverse transcription-polymerase chain reaction (RT-PCR) results showed that compared with Sham group, miR-22 expression level in myocardial tissues of rats decreased significantly in MI group. Overexpression of miR-22 could remarkably relieve cardiac insufficiency in MI rats, increase EF% and FS%, and reduce collagen deposition and the mRNA expression level of fibrosis-promoting genes in myocardial tissues of MI rats. The cross-sectional area of myocardial cells in MI + miR-22 mimic group was smaller than that in MI group. According to the results of immunohistochemical staining, overexpression of miR-22 notably reduced the level of oxidative stress marker 4-HNE in myocardial tissues of MI rats. Meanwhile, myocardial cells in MI + miR-22 mimic group exhibited a prominently lower apoptosis rate than those in MI group. Furthermore, Western blotting results demonstrated that overexpression of miR-22 inhibited the activation of the phosphatase and tensin homolog deleted on chromosome ten (PTEN)/Akt/mTOR signaling pathway in myocardial tissues of MI rats. The inhibitory effects of miR-22 on myocardial fibrosis and hypertrophy after MI in rats may be related to its inhibition on the PTEN/Akt/mTOR signaling pathway. All our findings suggested that miR-22 is expected to become a targeted drug for the clinical treatment of MI.

Association Between PD-L1 status Detected by SP142 and SP263 Antibody Clones and Clinical and Morphological Features of Gastric Cancer Factors

The aim is to study the relationship of PD-L1 status of SP142 and SP263 clones in gastric cancer with its clinical and morphological parameters. Material and methods. Surgical material obtained from 131 patients with a confirmed diagnosis of stomach cancer. Antibodies to PD-L1 SP263 were stained 127 cases, antibodies to PD-L1 SP142 – 126 cases. Reactions with PD-L1 were performed using the Ventana BenchMark Ultra device (Roche Ventana, USA) according to the protocols recommended by the manufacturer of primary antibodies. The results of immunohistochemical staining were compared with the clinical and morphological characteristics of gastric cancer using statistical analysis. Results. The positive PD-L1 status detected by clones SP263 and SP142 is significantly associated with type 2 macroscopic form according to the R. Bormann classification (p=0,003/p=0,003), tubular morphological type according to the WHO 5th edition, 2019 classification (p=0,001/p=0,018), intermediate morphological type according to the classification of P. Lauren (p=0,027/p=0,005) and the absence of signet ring cells (p=0,001/p=0,010). Staining with both clones used was not associated with the gender and age of patients, the size and localization of the tumor, the degree of its differentiation, the presence or absence of emboli in the lumen of blood and lymph vessels, the presence of affected lymph nodes and distant metastases, as well as the clinical stage (p&gt;0,05). The largest proportion of PD-L1 positive cases in the study of SP263/SP142 clones was found in tubular type tumors (75,6%/84,6%) according to the WHO 5th edition, 2019 classification, intermediate type (60,0%/68,0%) according to P. Lauren, type 2 (65,4%/68,0%) according to R. Bormann and in the absence of signet ring cells (70,7%/76,9%). Conclusion. Statistically significant clinical and morphological parameters of gastric cancer, where a reliable relationship between them and the expression of PD-L1 clones SP263 and SP142 was revealed, are: type 2 macroscopic form according to R. Bormann (p=0,003/p=0,003), tubular morphological type according to the classification of WHO 5th edition, 2019) (p=0,001/p=0,018), intermediate type according to P. Lauren (p=0,027/p=0,005) and the absence of signet ring cells (p=0,001/p=0,010).

Kv 11.1 Expression Is Associated With Malignancy of Canine Mammary Gland Tumors.

The expression level of the voltage-dependent potassium channel Kv 11.1 was shown to be associated with the clinicopathological features, aggressiveness, and prognosis of human breast cancer. Canine mammary gland tumor (cMGT) is the most common tumor type in intact female dogs; however, the significance of Kv 11.1 in cMGT is unknown. The aim of this study was to identify Kv 11.1 expression in 57 benign and malignant cMGT tissues from dogs and to investigate the correlation of Kv 11.1 expression with the clinicopathological parameters and prognosis of cMGT. A total of 57 samples were collected from cMGTs surgically resected at the Veterinary Medical Teaching Hospital, Seoul National University and subjected to immunohistochemistry assay using rabbit anti-Kv 11.1 polyclonal antibody. Immunohistochemical staining results were evaluated as the sum of intensity and percentage scores. The correlation between immunohistochemistry scores and clinicopathological parameters was investigated. Immunohistochemical analysis revealed that Kv 11.1 immunoreactivity was higher in benign cMGTs than in malignant cMGTs. Kv 11.1 expression was significantly associated with tumor malignancy (p<0.001), tumor size (p<0.001), histological grade (p<0.05), and age at the time of mastectomy (p<0.05). This study presents the first evidence of Kv 11.1 expression in cMGTs and indicates an inverse correlation between Kv 11.1 expression and tumor malignancy. Kv 11.1 expression can be used as a prognostic biomarker and a tool for the management of cMGTs.

Sinonasal Burkitt’s CD20 Negative Lymphoma : A Case Report

Burkitt's lymphoma is a B Cells non-lymphoma Hodgkin's that is aggressive, has a high degree of gradation, and is made up of diffuse, non-cleaved, undifferentiated, tiny cells that usually expresses CD20 positive . A very uncommon kind of juvenile cancer is Burkitt lymphoma (LB). 1, 2 100 new cases are recorded annually in the United States, compared to a frequency of 100 cases per million children in Africa. We report this case because of the rarity of Burkitt’s CD20 negative lymphoma. A 9-year-old girl who has a main complaint of eye swollen for almost two months. A bump with redness, swelling, and discomfort in the left eye that radiates to the head occurs after two weeks of before entering the hospital. Another complaint made by the patient was a big, pus-oozing lump in the left cheek and nose. On MSCT Scan examination, a solid mass was found in the left maxillary sinus, left ethmoid sinus, right left frontal sinus, left sphenoid sinus, left nasal cavity, and entered the extra orbital and left intraconal cavities, which forced the bulbus oculi to anterosuperior and caused proptosis bulbi. The mass also extended to the left parasella and caused destruction of the medial, anterior, and lateral walls of the left maxillary sinus. the patient will be treated with chemotherapy with the CHOP regimen with a dose of Cyclophosphamide 600 mg/m2 per body surface area (LPT), Doxorubicin 50 mg/m2 per LPT, Oncovin/Vincristine 1.4 mg/m2 per LPT, and Prednisone 50 mg. So the patient received Cyclophophamide 468 mg, Doxorubicin 40 mg, and Oncovin/Vincristine 1 mg. Then the patient was admitted to undergo the first chemotherapy. In this case it was suffered by a 9 year old girl and the symptoms of a lump in the eye and left cheek were getting bigger and bigger, with histopathological results obtained cells with "Starrys Sky" pattern, which supports the diagnosis of Burkitt's Lymphoma so that the patient receives chemotherapy treatment. The result of immunohistochemical staining for CD20 was negative.The chemotherapy regimen consist of cychlosfamide, doxorubicin, oncovin and prednisone. Patient was diagnosed with stage II E Burkitt lymphoma, ECOG I, ccomplete respons.

Monomorphic Epitheliotropic Intestinal T-Cell Lymphoma:Pathological Phenotype and Genetic Mutation Analysis of 12 Patients

Introduction: Monomorphic Epitheliotropic Intestinal T-Cell Lymphoma (MEITL) is a primary intestinal lymphoma. Previously, it was referred to as Enteropathy Associated T-cell Lymphoma (EATL) type II. However, in the 2016 revision of the WHO classification, MEITL was recognized as a separate entity independent of EATL. EATL is mainly observed in people of Nordic descent and is closely associated with celiac disease, whereas MEITL is more commonly found in Asians and Hispanics without a history of celiac disease. MEITL is an extremely rare condition, accounting for approximately 5% of all Peripheral T-cell Lymphomas (PTCLs). It predominantly affects the small intestine, especially the jejunum. The tumor cells often display an activated cytotoxic T-cell phenotype and express CD8 and CD56. Due to its rarity, there is limited understanding of this disease. Between 2017 and 2022, our center diagnosed only 14 cases of MEITL. We analyzed the clinical characteristics and pathological phenotypes of these patients and further investigated the status of gene somatic mutations using a panel-based genomic exome analysis. Results: Clinical features: From the medical records system, a total of 622 gastrointestinal live tissue biopsy samples were searched from October 2017 to September 2022. Among them, 67 cases (10.8%) were diagnosed as PTCLs, with only 15 cases (2.4%) being identified as MEITL, accounting for 22.3% of all gastrointestinal PTCLs. Out of the 15 MEITL cases, 9 were male, and 6 were female, with a median age of 61 years(figure). Treatment and Efficacy: Seven patients underwent elective surgery due to the involvement of malignant tumors in the gastrointestinal tract, while six patients received emergency surgery for intestinal perforation. Three patients did not undergo chemotherapy, and the remaining 12 patients received a median of 5 (range 1-8) cycles of chemotherapy using the CHOP/CHOP-like regimen. For elderly and frail patients, the dosage was reduced by 20% to 50%. After treatment, one patient achieved complete remission (CR/CRu), three patients had a partial response (PR), six patients had stable disease (SD), and two patients had uncontrolled disease (PD). Pathological Characteristics: The diagnosis of MEITL was confirmed by two hematopathologists. The immunohistochemical staining results are shown in Figure. The results revealed that all patients expressed CD3, CD8, and CD56. Only four cases expressed CD4, including three cases with partial expression. TIA1 was expressed in 11 patients, and granzyme B was expressed in 11 patients, with eight cases showing co-expression of TIA1 and granzyme B. None of the patients expressed CD20, while one case showed weak expression of CD30. Ki67 expression was observed in all cases, with seven cases having Ki67 expression above 80% and all others showing expression above 50%. Gene Mutations: We conducted the Next-Generation Sequencing in 12 patients. The genes include STAT5B (9 cases), SETD2 (7 cases), NRAS (3 cases), KRAS (4 cases), JAK3 (4 cases), BRAF (1 case), BCOR (1 case), CREBBP (4 cases), EZH2 (1 case), EP300 (1 case), JAK1 (1 case), TP53 (3 cases), ATM (1 case), SOCS1 (1 case), CHD2 (1 case), and JAK2 (1 case). All patients showed abnormalities in two or more genes. Survival: Median follow-up time was 4.45 months, and among 15 patients, 14 experienced disease progression. The median progression-free survival (PFS) was 2.9 months (95% CI: 1.7 months to 4.2 months). Up to the follow-up cutoff, 11 patients had died, and the median overall survival (OS) was 4.8 months (95% CI: 3.2 months to 6.4 months). Patients who underwent early elective surgery showed a significant improvement in OS (P=0.012), with a median OS of 18.0 months. Based on gene mutation analysis, mutations in STAT5B and RAS (NRAS or KRAS) (P=0.038, 0.007) were associated with poorer OS, whereas abnormalities in genes like SETD2 did not affect patient OS. Conclusion: Our results show that MEITL is extremely rare, accounting for only 2.4% of all lymphomas involving the intestines. Patients with MEITL have a poor prognosis, with a median OS of less than six months. However, early elective surgery can significantly improve patient survival. MEITL is characterized by a cytotoxic T-cell phenotype, with the majority expressing TIA1 and granzyme B. Commonly occurring gene mutations are found in STAT5B, SETD2, and NRAS or KRAS. Mutations in STAT5B and RAS genes (NRAS or KRAS) often indicate a shorter OS.