Presence of Merkel cell polyomavirus DNA and large-T antigen in keratinocyte carcinomas and its correlation with immunohistochemical markers p16, p53 and ki67

Abstract Background Merkel cell polyomavirus (MCPyV), a human polyomavirus that is unequivocally linked to merkel cell carcinoma (MCC), has been found in association with keratinocytes carcinomas (KC), especially basal cell carcinoma (BCC) and cutaneous squamous cell carcinoma (cSCC). Nevertheless, there is scarce information about the possible involvement of MCPyV in the development of KC. Objectives To assess the presence of MCPyV DNA and Large-T Antigen (LT-Ag) via Polymerase Chain Reaction (PCR) and Immunohistochemistry (IHC) in cases of KC, and to correlate its presence with immunohistochemical markers p16, p53, and ki67, tumor type and subtype, sun-exposed location, and epidemiological data. Methods The prevalence of MCPyV DNA, LT-Ag, and immunohistochemical markers p16, p53, and ki67 was assessed by PCR and Immunohistochemistry (IHC) in 127 cases of KC, these results were correlated with tumor type and subtype, sun-exposed location, and epidemiological data. Results The MCPyV DNA was detected in 42.57% (43 of 101) cases by PCR, the LT-Ag was detected in 16.4% (20 of 122) of cases, p16 in 81.5% (97 of 119), p53 in 66.4% (83 of 125), ki67 in 89% (73 of 82). No correlation between MCPyV LT-Ag and DNA confronted with tumor type, subtype, location site, and immunohistochemical markers was found. A single correlation between the MCPyV LT-Ag and cSCC tumors and peri-tumoral lymphocyte cells was noted. Study limitations Further steps need to be taken to better evaluate the MCPyV influence and its possible role in KC carcinogenesis, as the evaluation of the virus genome state, the gene sequence that encodes LT-Ag in the KC tumor cells, and in situ hybridization for viral DNA or RNA in these cells. Conclusions Despite the frequent detection of MCPyV in KC, the data available so far does not support the hypothesis of a causal relationship between them.

Elevada expressão de galectina-3 em cicatrizes excessivas: Um estudo piloto / Elevated expression of galectin-3 in excessive scars: A pilot study

Introdução: Cicatrizes hipertróficas e queloides são tipos de cicatrizes excessivas de cicatrização anormal da pele. Galectina-3 (gal-3) é uma proteína da família das lectinas capaz de identificar carboidratos, que podem se combinar e atuar em diversas moléculas. Na literatura, a ação da gal-3 como principal agente regulador da fibrogênese já foi descrita, sendo atualmente utilizada na terapia antifibrótica de diversos órgãos como pulmão e fígado. O objetivo deste estudo piloto foi mostrar resultados preliminares encontrados na expressão de gal-3 em cicatrizes exacerbadas. Método: Foram coletadas 20 amostras de biópsias de cicatrizes excessivas (16 queloides e 4 cicatrizes hipertróficas) e 9 amostras de cicatrizes normais de 22 mulheres e 7 homens. Essas amostras foram processadas para análise histopatológica de rotina por imuno-histoquímica para detectar gal-3. As células positivas para gal-3 foram quantificadas pelo método estereológico utilizando uma grade de 36 pontos. Resultados: A imuno-histoquímica mostrou alta expressão de gal-3 em células endoteliais e epiteliais de todas as amostras de cicatrizes, bem como expressão em células distribuídas pela derme. Maior expressão de gal-3 foi encontrada em amostras de queloides (28% de células positivas) em comparação com cicatrizes normais (18%) e hipertróficas (22%) (p=0,0075). Os resultados foram obtidos de um pequeno número de pacientes, por se tratar de um estudo piloto. Conclusão: Os dados sugerem que a gal-3 participa do processo de cicatrização e, devido à sua maior presença em amostras de queloides, pode ser um potencial biomarcador para formação de queloides e um alvo terapêutico promissor a ser explorado.

Introduction: Hypertrophic scars and keloids are types of excessive scars from abnormal skin healing. Galectin-3 (gal-3) is a protein from the lectin family capable of identifying carbohydrates, which can combine and act on different molecules. In the literature, the action of gal-3 as the main regulatory agent of fibrogenesis has already been described and is currently used in anti-fibrotic therapy for various organs such as the lung and liver. The objective of this pilot study was to show preliminary results found in the expression of gal-3 in exacerbated scars. Method: Twenty biopsy samples from excessive scars (16 keloids and 4 hypertrophic scars) and 9 samples from normal scars were collected from 22 women and 7 men. These samples were processed for routine histopathological analysis by immunohistochemistry to detect gal-3. Gal-3 positive cells were quantified by the stereological method using a 36-point grid. Results: Immunohistochemistry showed high expression of gal-3 in endothelial and epithelial cells of all scar samples, as well as expression in cells distributed throughout the dermis. Higher gal-3 expression was found in keloid samples (28% positive cells) compared to normal (18%) and hypertrophic (22%) scars (p=0.0075). The results were obtained from a small number of patients, as this was a pilot study. Conclusion: The data suggest that gal-3 participates in the healing process and, due to its greater presence in keloid samples, it may be a potential biomarker for keloid formation and a promising therapeutic target to be explored.

Comparative study of fine needle aspiration cytology and tru-cut biopsy in the diagnosis of breast lesions

Background: Breast carcinoma is one of the most researched cancers across the world. FNAC is often used as a first priority investigation in patients with breast lump. Tru-cut biopsy is useful in preoperative knowledge of prognostic parameters with the help of IHC markers ER, PR, Her2 neu. The technique is reliable, simple and reproducible which can be used even in resource poor countries like India. To analyze the sensitivity, specificity, positive predictive values and negative predictive values and the diagnostic accuracy (efficacy) of fine needle aspiration cytology and trucut biopsy. Methods: The study included 82 patients presenting with palpable breast lesions. All patients underwent FNAC followed by Tru-cut biopsy under ultrasound guidance. ER, PR, HER2 Neu, cytokeratin immunostaining was done in malignant cases. The data collected analysed statistically. Results: The sensitivity, specificity, PPV, NPV of FNAC were 96.61 %, 95.65 %, 98.28%, 90.91% respectively and that of trucut biopsy were 100%, 100%, 100%, and 100 % respectively. Conclusions: Out of total 82 patients, maximum no of cases (57) were duct carcinoma accounting 69.50% on FNAC and Tru-cut biopsy. IHC was done in all carcinomas. Maximum (46 cases) were luminal type. The present study emphasizes that tru-cut biopsy can serve as confirmative diagnostic tool over FNAC and provides information regarding prognostic factors and treatment modalities based on IHC markers.

Assessing the diagnostic impact of P63, PSA and BCL-2 proteins in premalignant and malignant prostate tissues

Background: Prostate cancer (CaP) is increasingly becoming a major health issue affecting men as cancer-related fatalities are attributable to the condition. Immunohistochemistry (IHC) diagnostic criteria can help in gene-targeted therapy and help reduce its prevalence. This study is to assess the diagnostic impact of prostate-specific antigen (PSA), P63 and BCL-2 antibodies in CaP. Method: A case-controlled retrospective study was carried out on eighty (80) prostrate tissue blocks retrieved from the pathology archive of Ekiti State university teaching hospital Ado Ekiti. IHC analysis of the selected antibodies was carried out and also stained with haematoxylin and eosin (H and E) for second opinion and confirmation. Results: The study showed that all the CaP samples had 100% positivity with varying reactivity to the IHC biomarkers; PSA had 100% positivity and MPR of 94% due to its multiple weaknesses as a biomarker p63 is a basal cells marker. Conclusions: The expressions of these antibodies were observed in the progression of CaP. Although these markers are useful in predicting the progression from benign prostatic hyperplasia (BPH) to CaP, none of them can be utilised in isolation to a conclusion. Hence, they should be used in conjunction with one another to make up for their limitations. The immunohistochemical markers are beneficial in CaP diagnosis.

Exploring neuroendocrine carcinoma of the cervix: a case report and literature review

Neuroendocrine neoplasms (NENs) are rare, comprising less than 1% of cervical malignancies. Diagnosis and management of cervical neuroendocrine carcinoma (NEC) pose challenges due to its rarity and aggressive nature. Aim of the study was to highlight the importance of considering NEC in the differential diagnosis of cervical malignancies and underscore the potential benefits of comprehensive treatment approaches. We report a case of a 35-year-old woman presenting with post-coital bleeding, subsequently evaluated and diagnosed with NEC T1b1N0Mx following radical hysterectomy. Adjuvant chemoradiotherapy and chemotherapy led to one year of remission. Cervical NEC poses diagnostic and therapeutic challenges due to its rarity and aggressive nature. Despite multimodal treatment, prognosis remains poor, warranting further research and targeted therapies. The rarity of cervical NEC emphasizes the need for increased awareness among clinicians to facilitate early detection and appropriate management. Further studies are warranted to explore novel treatment modalities and improve outcomes for patients with this challenging malignancy.

Alpaca (Vicugna pacos) Skin Analyzed Under Architecture and Structure / Piel de Alpaca (Vicugna pacos) Analizada Bajo Arquitectura y Estructura

SUMMARY: This study evaluated the morphology of alpacas skin. Biopsies were collected and samples were fixed in 10 % neutral buffered formalin for histological procedures. The sections were stained with hematoxylin and eosin, picrosirius red and Masson's trichrome. Types I, III and IV collagen were analyzed by immunohistochemistry. The derma presented sebaceous and sweat glands, as well as follicular groups with medullated fibers. Type I and type IV collagen were observed at epidermis and dermis as well as in glandular structures and hair follicles. The collagen III, was observed only in dermis.

Este estudio evaluó la morfología de la piel de alpacas. Se recogieron biopsias y las muestras se fijaron en formalina tamponada neutra al 10 % para procedimientos histológicos. Las secciones se tiñeron con hematoxilina y eosina, rojo picrosirius y tricrómico de Masson. El colágeno tipo I, III y IV se analizó mediante inmunohistoquímica. La dermis presentó glándulas sebáceas y sudoríparas, así como grupos foliculares con fibras medulares. Se observó colágeno tipo I y tipo IV en la epidermis y la dermis, así como en estructuras glandulares y folículos pilosos. El colágeno III, se observó únicamente en la dermis.

Immunohistochemical expression of SNAIL and SLUG in endometrioid endometrial cancer and precursor lesions

Background: Endometrioid endometrial carcinoma is one of the most prevalent gynecologic cancers. The epithelial-mesenchymal transition is a multistep process involved in the development of cells that results in disappearance of intercellular adhesion in the epithelium and acquiring mesenchymal properties, hence influence cancer progression and metastasis. Loss of intercellular adhesion can be activated by different mechanisms, including transcriptional repression. However, the expression of transcriptional repressors in EEC and precursor lesions remain to be investigated. In the present study, we evaluated the immunohistochemical expression of proteins of transcriptional after repression snail and slug in EEC and its precursor lesions.Methods: It was a prospective nested case-control study on women from 35 to 70 years of age. Endometrial biopsies were obtained and processed for routine histological diagnosis. Immunohistochemical expression of snail and slug were evaluated.Results: A total of 39 EEC cases, 37 Endometrial hyperplasia and 19 normal controls were included in this study. Expression of snail was positive in 77% (30/39) and slug in 82% (32/39) of EEC cases while in precancer group snail was positive in 76% (28/37) and slug in 68% (25/37). In normal control snail was positive in 32% (6/19) and SLUG in 21% (4/19).Conclusion: Up-regulation of snail & slug observed in both precancer and cancer cases, suggesting their involvement from an early stage of carcinogenesis. Therefore therapies targeted at transcriptional repressors at an early stage of carcinogenesis, i.e., at precancerous lesions, could play a valuable role in reducing cancer progression and metastasis.

Nephroprotective effect of the methanolic fruit extract of Musa balbisiana Colla against carbon tetrachloride-induced toxicity in Swiss albino mice

Renal failure is the loss of renal functions owing to multiple factors, including oxidative stress, and it is a global concern, as it costs millions of lives every year. Carbon tetrachloride is a standard toxicant exploited to induce nephrotoxicity in experimental animals to study oxidative stress-related renal injury. This study was conducted to determine the nephroprotective activity of a methanolic extract derived from the unripe fruit pulp of Musa balbisiana Colla (MBME) in carbon tetrachloride-induced toxicity in Swiss albino mice. Multiple groups of mice were treated with a single dose of carbon tetrachloride (1-mg/kg body weight) intraperitoneally once in a week followed by oral administration of the methanolic fruit extract of M. balbisiana Colla (MBME) at two different doses (200 and 400 mg/kg body weight) consecutively for 28 days. Treatment with carbon tetrachloride reduced body weight along with enzymatic and non-enzymatic antioxidants in the renal tissue, elevated kidney weight, renal biomarkers, lipid peroxidation, IL-6, TNF-?, and TGF-? levels in the blood serum, impaired antioxidant system and up-regulated TGF-? expression in the kidney tissue, and brought histological changes in the architecture of renal tissues. Our data revealed that the oral administration of the methanolic fruit extract of M. balbisiana Colla (MBME) dose wisely restored kidney and body weight, renal biomarkers, enzymatic and non-enzymatic antioxidants in the renal tissue, reduced lipid peroxidation, interleukin 6 (IL-6), tumor necrosis factor-alpha (TNF-?), and transforming growth factor-? (TGF-?) levels in the blood serum, re-established antioxidant system and down-regulated TGF-? expression in the kidney tissue, and re-established the architecture of renal tissues. Thus, this study reported the nephroprotective effects of methanolic fruit extract of M. balbisiana Colla (MBME) in carbon tetrachloride-induced toxicity in Swiss albino mice through possible antioxidant and anti-inflammatory activity.

CLCA1 is Poorly Expressed in Stomach Adenocarcinoma and Correlates with Immune Infiltration / CLCA1 se Expresa Deficientemente en el Adenocarcinoma de Estómago y se Correlaciona con la Infiltración Inmune

SUMMARY: Calcium-activated chloride channel regulator 1 (CLCA1) is associated with cancer progression. The expression and immunologic function of CLCA1 in stomach adenocarcinoma (STAD) remain unclear. In this investigation, the expression of CLCA1 in STAD tissues and its involvement in the progression and immune response of STAD were examined using databases such as cBioPortal, TISIDB, and UALCAN. In order to validate the expression level of CLCA1 protein in gastric adenocarcinoma, thirty clinical tissue specimens were gathered for immunohistochemical staining. The findings indicated a downregulation of CLCA1 in STAD patients, which was correlated with race, age, cancer grade, Helicobacter pylori infection, and molecular subtype. Through the examination of survival analysis, it was identified that diminished levels of CLCA1 within gastric cancer cases were linked to decreased periods of post-progression survival (PPS), overall survival (OS), and first progression (FP) (P<0.05). The CLCA1 mutation rate was lower in STAD, but the survival rate was higher in the variant group. The correlation between the expression level of CLCA1 and the levels of immune infiltrating cells in STAD, as well as the immune activating molecules, immunosuppressive molecules, MHC molecules, chemokines, and their receptor molecules, was observed. Gene enrichment analysis revealed that CLCA1 may be involved in STAD progression through systemic lupus erythematosus (SLE), proteasome, cell cycle, pancreatic secretion, and PPAR signaling pathways. In summary, CLCA1 is anticipated to function as a prognostic marker for patients with STAD and is linked to the immunization of STAD.

El regulador 1 del canal de cloruro activado por calcio (CLCA1) está asociado con la progresión del cáncer. La expresión y la función inmunológica de CLCA1 en el adenocarcinoma de estómago (STAD) aún no están claras. En esta investigación, se examinó la expresión de CLCA1 en tejidos STAD y su participación en la progresión y respuesta inmune de STAD utilizando bases de datos como cBioPortal, TISIDB y UALCAN. Para validar el nivel de expresión de la proteína CLCA1 en el adenocarcinoma gástrico, se recolectaron treinta muestras de tejido clínico para tinción inmunohistoquímica. Los hallazgos indicaron una regulación negativa de CLCA1 en pacientes con STAD, que se correlacionó con la raza, la edad, el grado del cáncer, la infección por Helicobacter pylori y el subtipo molecular. Mediante el examen del análisis de supervivencia, se identificó que los niveles reducidos de CLCA1 en los casos de cáncer gástrico estaban relacionados con períodos reducidos de supervivencia posterior a la progresión (PPS), supervivencia general (OS) y primera progresión (FP) (P <0,05). La tasa de mutación CLCA1 fue menor en STAD, pero la tasa de supervivencia fue mayor en el grupo variante. Se observó la correlación entre el nivel de expresión de CLCA1 y los niveles de células inmunes infiltrantes en STAD, así como las moléculas activadoras inmunes, moléculas inmunosupresoras, moléculas MHC, quimiocinas y sus moléculas receptoras. El análisis de enriquecimiento genético reveló que CLCA1 puede estar involucrado en la progresión de STAD a través del lupus eritematoso sistémico (LES), el proteasoma, el ciclo celular, la secreción pancreática y las vías de señalización de PPAR. En resumen, se prevé que CLCA1 funcione como un marcador de pronóstico para pacientes con STAD y está vinculado a la inmunización de STAD.

A cutaneous malignant granular cell tumour: an uncommon entity with diagnostic challenge

Granular cell tumour (GCT) is rare and accounts for approximately 0.5% of all soft tissue tumours. The malignant GCT (MGCT) especially cutaneous malignant granular cell tumour is extremely rare constituting 1-2% of all granular cell tumours and mostly found in the subcutaneous soft tissues of lower exrtremities, especially thighs. The uncommon occurrence of cutaneous MGCT and their histopathological similarities with other entities make diagnosis difficult in some cases. Here we report a case of 36 years old male patient who presented with a mass in the skin of right lower abdominal wall which has been increased gradually over the last one year without pain. The size of the mass is approximately 6.5 cm in greatest dimension, firm in consistency with surface irregularity and ulceration diagnosed as malignant GCT at the histopathological examination showing focal ulceration and lined by keratinized stratified squamous epithelium revealing acanthosis and pseudoepitheliomatous hyperplasia. The dermis show neoplastic epithelioid cells arranged in sheets and nests with vesicular chromatin, conspicuous to prominent nucleoli, and abundant amount of fine granular eosinophillic cytoplasm. Mitosis is more than 2/10HP. Immunohistochemical stains for S-100, CD 68 and vimentin were positive in the lesional cells.

IL-1β and IL-17 in cutaneous lupus erythematous skin biopsies: could immunohistochemicals indicate a tendency towards systemic involvement?

Abstract Background: Only a fraction of patients with cutaneous lupus erythematosus (CLE) will eventually progress toward systemic disease (SLE). Objective: To find inflammatory biomarkers which could predict the progression of cutaneous lupus erythematosus (CLE) into systemic lupus erythematosus (SLE) using immunohistochemical (IHC) assays. Methods: Immunohistochemical markers for cytotoxic, inflammatory, and anti-inflammatory responses and morphometric methods were applied to routine paraffin sections of skin biopsies, taken from lesions of 59 patients with discoid lupus, subacute lupus, and lupus tumidus. For the diagnosis of SLE, patients were classified by both the American College of Rheumatology (ACR-82) and the Systemic Lupus International Collaborating Clinics (SLICC-12) systems. Results: Skin samples from CLE/SLE +patients presented higher expression of IL-1β (ARC-82: p = 0.024; SLICC-12: p = 0.0143) and a significantly higher number of cells marked with granzyme B and perforin (ARC: p = 0.0097; SLICC-12: p = 0.0148). Biopsies from CLE/SLE- individuals had higher expression of IL-17 (ARC-82: p = 0.0003; SLICC-12: p = 0.0351) and presented a positive correlation between the density of granzyme A+and FoxP3+ cells (ARC-82: p = 0.0257; SLICC-12: p = 0.0285) and CD8+ cells (ARC-82: p = 0.0075; SLICC-12: p = 0.0102), as well as between granulysin-positive and CD8+ cells (ARC-82: p = 0.0024; SLICC-12: p = 0.0116). Study limitations: Patients were evaluated at a specific point in their evolution and according to the presence or not of systemic disease. The authors cannot predict how many more, from each group, would have evolved towards SLE in the following years. Conclusions: In this cohort, immunohistochemical findings suggested that patients with a tendency to systemic disease will show strong reactivity for IL-1β, while those with purely cutaneous involvement will tend to express IL-17 more intensely.

Tumor maligno de la vaina nerviosa periférica: reporte de un caso / Malignant peripheral nerve sheath tumor, a case report

Resumen El tumor maligno de la vaina nerviosa periférica (TMVNP) es un sarcoma de alto grado de malignidad. Es poco frecuente, agresivo y generalmente se localiza en tronco y miembros inferiores. Se presenta mayormente en pacientes con neurofibormatosis tipo 1, aunque no siempre se encuentra esta asociación. Este tumorcomparte características histológicas e inmunohistoquímicas con el melanoma, lo que puede dificultar el diagnóstico. Presentamos el caso de un paciente con TMVNP, en el cual los hallazgos histológicos iniciales condujeron a un diagnósticoerróneode melanoma.

Abstract Malignant peripheral nerve sheath tumor (MPNST) is a high-grade sarcoma. It is rare, aggressive and generally located on the trunk and lower limbs. It occurs in a high percentage of patients with neurofibormatosis type 1, although this association is not always found. This tumor shares histological and immunohistochemical characteristics with melanoma, which can make diagnosis difficult. We present the case of a patient with MPNST, in whom the initial histological findings led to an erroneous diagnosis of melanoma.

Schwannoma de colon derecho: a propósito de un caso / Schwannoma of the right colon: case report

RESUMEN El schwannoma de colon es una entidad sumamente rara que puede debutar como lesión subepitelial con signos ulcerativos de melena y anemia. El estudio de imágenes nos orienta a la localización mientras que la biopsia colonoscópica no es de ayuda. Muchas veces el diagnóstico y tratamiento se efectúa con la resección de la lesión en tanto que el diagnóstico final se realiza en el posoperatorio por histopatología y por la inmunohistoquímica, la cual muestra positividad intensa para S100 y vimentina en las células tumorales con un índice de proliferación KI67 menor al 1%, por lo que se concluye que se trata de una lesión benigna. Presentamos el siguiente caso por su dificultad diagnóstica pre e intraoperatoria, clínica inespecífica y diagnóstico definitivo por inmunohistoquímica.

ABSTRACT Colon schwannoma is an extremely rare entity that may debut as a subepithelial lesion with ulceration signs, such as melena and anemia. Imaging studies guide us to localization, while a colonoscopy biopsy is not helpful. Many times, the diagnosis and treatment are made with lesion resection, and the final diagnosis is postoperatively made with histopathology and immunohistochemistry, which shows intense positivity for S100 and vimentin in tumor cells with a KI67 proliferation index of less than 1%, therefore, it is concluded that this is a benign lesion. We present this case due to its pre- and intraoperative diagnostic difficulty, non-specific symptoms, and its definitive diagnosis that was achieved with immunohistochemistry.

Relationship between zinc finger protein A20, CTGF and FibroScan and fibrosis in chronic hepatitis B patients / 公共卫生与预防医学

Objective Studies on the expression and location of zinc finger protein A20 (A20) and connective tissue growth factor (CTGF) in liver tissues of patients with chronic hepatitis B were conducted, and the relationship between them and liver fibrosis was determined by FibroScan. Methods Studies on A20 and CTGF in liver tissues of 160 patients with chronic hepatitis B were conducted in accordance with the stage of pathological fibrosis and inflammation of the liver, and quantitative immunohistochemistry test was conducted, and statistical analysis was conducted by FibroScan. Results The expressions of A20 and CTGF in liver tissues increased with the aggravation of liver pathological fibrosis and inflammation, and there were significant differences between each stage and the control group (P0.05). There was positive correlation between liver A20 and CTGF, r=0.796 (P<0.05). Conclusions In patients with chronic hepatitis B, A20, CTGF and FibroScan are positively correlated with the degree of liver fibrosis, and A20 and CTGF are also positively correlated with the degree of liver inflammation, which can be used as indicators to evaluate the degree of liver inflammation and fibrosis, and further guide the anti-inflammatory and anti-fibrosis treatment of patients.

Expression and significance of cyclin dependent kinase inhibitor 3 in HPV16-positive cervical cancer / 重庆医学

Objective To investigate the expression and significance of cyclin dependent kinase inhibitor 3(CDKN3)in human papillomavirus type 16(HPV16)-positive cervical cancer.Methods CDKN3 expression in pan-cancer was retrieved and downloaded from the Gene Expression Profiling Interactive Analysis(GEPIA)platform,and the expression levels of CDKN3 between normal cervical tissues(13 samples)and cervical cancer tissues(306 samples)were compared.Subsequently,GSE39001 data of HPV16-positive cervical cancer was sourced and downloaded from the Gene Expression Omnibus(GEO)database,and the expression levels of CDKN3 mRNA in HPV16-positive cervical cancer tissues(43 samples)and normal cervical tissues(12 samples)were compared.Immunohistochemical method was used to detect the expression of CDKN3 in 12 ca-ses of HPV16-positive cervical cancer,12 cases of HPV16-positive cervical precancerous lesions,10 cases of HPV16-positive chronic cervicitis and 7 cases of HPV-negative normal cervical samples collected from the Af-filiated Hospital of Guizhou Medical University.SiHa(HPV16-positive),HeLa(HPV18-positive)and HCC94(HPV-negative)cervical cancer cell lines were selected,and their CDKN3 expression were detected by West-ern blot.Results The GEPIA platform analysis showed that CDKN3 was highly expressed in pan-cancer,and the expression level of CDKN3 in cervical cancer tissue was significantly higher than that in normal cervical tissue(P<0.05).The GEO dataset reflected a significantly increased CDKN3 mRNA expression level in HPV16-positive cervical cancer compared to normal cervical tissue(P<0.001).Immunohistochemical verifi-cation showed that the positive expression rates of CDKN3 in HPV16-positive cervical cancer,HPV16-positive cervical precancerous lesion,HPV16-positive chronic cervicitis and HPV-negative normal cervical tissues were 91.7%,58.3%,0 and 0,respectively.Western blot analysis of cervical cancer cells showed that the expression level of CDKN3 in SiHa(HPV16-positive)cells was significantly higher than that in HeLa(HPV18-positive)and HCC94(HPV-negative)cells(P<0.05).Conclusion CDKN3 is a new oncogene of HPV16-positive cer-vical cancer,which may be used as a marker of cervical precancerous lesions and cervical cancer screening,and may provide a theoretical basis for subsequent mechanism research and targeted therapy.

Mechanism of peimine improving chronic obstructive pulmonary disease induced by lipopolysaccharide combined with cigarette smoke in mice / 解剖学报

Objective To investigate the effects and mechanisms of peimine(PME)on chronic obstructive pulmonary disease(COPD)in mice.Methods The mice were randomly divided into 4 groups(20 mice in each group),control group,PME group,chronic obstructive pulmonary disease group and treatment group.Animal models of COPD were induced in mice by lipopolysaccharide combined with smoke.The effects of PME on COPD model mice was analyzed by HE staining,transmission electron microscopy and the ratio of wet/dry weight of mouse lung tissue.The effects of PME on COPD model mice were analyzed by HE staining,transmission electron microscopy and the ratio of wet/dry weight of mouse lung tissue.The effects of PME on inflammatory factor tumor necrosis factor(TNF)-α,interleukin(IL)-6 and IL-1β in lung tissue were analyzed by ELISA and Western blotting.The effects of PME on oxidative stress in lung tissue were analyzed by dihydroethidium(DHE)staining and Western blotting.The effects of PME on nuclear factor kappa-B(NF-κB)pathway and nuclear factor erythroid 2-related factor 2(Nrf2)pathway were analyzed by protein immunoblotting.Results Compared with the COPD group,PME treatment could significantly improve the lung tissue injury and the number of inflammatory cells in mice,and the wet/dry weight ratio of lung tissue was significantly reduced.Compared with the control group,the levels of TNF-α,IL-6 and IL-1β in the alveolar lavage fluid of COPD mice significantly increased,and the level of TNF-α,IL-6 and IL-1β in the alveolar lavage fluid of mice after PME treatment was significantly reduced.In addition,compared with the control group,the protein expression of TNF-α,IL-6 and IL-1β in the lung tissue of COPD mice significantly increased,and the level of TNF-α,IL-6 and IL-1β in the lung tissue of COPD mice after PME treatment were significantly reduced.Immunohistochemistry and Western blotting showed that the level of superoxide dismutase 2(SOD2)protein in COPD group was significantly lower than that in control group,while PME treatment could improve the level of superoxide dismutase protein.The analysis of MDA content in lung tissue showed that compared with the COPD group,the production of MDA in lung tissue of COPD mice was significantly inhibited after PME treatment.Protein Western blotting showed that PME treatment could prevent the phosphorylation of inhibitor of NF-κB(IκBα)and the transfer of NF-κB p65 to the cell nucleus,and the expression of Nrf2 and its main downstream target heme oxygenase-1(HO-1)in the lung tissue of mice treated with PME significantly increased.Conclusion PME is able to inhibit inflammation and oxidative stress and improve lung tissues damage in the COPD model in vivo and this protection effect might be both the Nrf2 pathway activation and NF-κB pathway inhibition.

Expression of CXCR5 and its significance of clinical and pathological in classical Hodgkin lymphoma / 临床与实验病理学杂志

Purpose To investigate the expression of Che-mokine(C-X-C Motif)receptor 5(CXCR5)and its clinico-pathological significance in classic Hodgkin lymphoma(CHL).Methods The expression of CXCR5 was assessed in 33 pa-tients by immunohistochemistry(IHC),and retrospectively ana-lyzed the expression and clinical significance of CXCR5 in the four subtypes of CHL.Meanwhile,10 cases of ALK-positive an-aplastic large cell lymphoma(ALCL)and 10 cases of ALK-neg-ative ALCL were collected as the control group.ResultsThere were 31 cases with CXCR5-positive in all 33 cases(93.94％),including 15/16(93.75％)in nodular sclerosis CHL,12/13(92.31％)in mixed cellularity CHL,2/2 in lymphocyte-rich CHL,and 2/2 in lymphocyte-depleted CHL.The positive ex-pressions of CXCR5 in different immunophenotypes of CHL were as follow,31/33(93.94％)in CD30 positive and PAX5 weakly positive CHL.12/14(85.71％)in CD15 negative CHL,24/26(92.31％)in CD20 negative CHL,10/11(90.91％)in EBER-negative CHL and 5/6 in LMP1-negative CHL.CXCR5 were not expressed in all 20 cases of ALCL.Conclusion The positive expression rate of CXCR5 in CHL is high.When the tumor cells are negative for CD15,LMP1 and CD20 or EBER,CXCR5 also has a high positive expression rate,which is helpful for the diagnosis of CHL.CXCR5 can be used to differentiate CHL from ALCL,especially the cases lacking typical morpholo-gy and immunohistochemistry.

Pathological characteristics of angioimmunoblastic T cell lymphoma with bone marrow involvement / 临床与实验病理学杂志

Purpose To explore the pathological features of angioimmunoblastic T-cell lymphoma(AITL)with bone marrow involvement and to improve awareness of bone marrow infiltration in AITL.Methods The tissue morphology of 32 cases of AITL with bone marrow involvement was retrospectively analyzed.Im-munohistochemistry using the EnVision method and ten-color flow cytometry were conducted to detect AITL-related immune markers.T cell clonality was analyzed through T cell receptor(TCR)gene rearrangement.Results The predominant pat-terns of tumor cell infiltration were nodular(20/32,62.5％)and interstitial or small clusters(10/32,31.3％).The nodules showed a mixture of cellular components.In some cases,the fo-ci contained a mixture of cells with characteristic"granuloma-toid"changes.The tumor cells were mainly small to medium-sized lymphocytes with inconspicuous atypia.Some cases showed plasma cell proliferation.19 cases were subject to immunohisto-chemical staining,which revealed a low count of CD4-positive T cells,with an average of 8.4％.The positive rates of T follic-ular helper cells(TFH)markers were as follows:CD10(7/14,50.0％),BCL6(6/19,31.6％),PD-1(13/19,68.4％),and CXCL13(13/19,68.4％).In most cases,tumor cells showed co-expression of PD-1 and CXCL13,but the number of positive cells was less than 1％.Flow cytometry analysis was performed in 24 cases,among which 22 cases all consistently expressed cytoplasmic CD3(cCD3),CD5,CD4,and CD2,with varying degrees of CD10 expression.In some cases,there was a lack of expression of surface CD3(sCD3)(12/22,54.5％),while there was a lack of expression of CD7(8/22,36.4％).and no abnormal T cells were found in 2 cases.TCR gene rearrangement analysis was performed in 7 cases,with 3 cases showing TCR clonality.Conclusion AITL with bone marrow involvement exhibits a lower proportion of tumor cells and less atypia,making it prone to misdiagnosis.The presence of lymphocytic foci with mixed cellular components in the bone marrow can indicate bone marrow involvement in AITL.Flow cy-tometry detection of abnormal T cells(double positive for CD4 and CD10)strongly suggests bone marrow infiltration in AITL.A comprehensive diagnosis of bone marrow involvement in AITL re-quires consideration of bone marrow biopsy,flow cytometry,and TCR gene rearrangement analysis.

Molecular characteristics of diffuse large B-cell lymphoma with high expression of MYC:analysis of 45 cases / 临床与实验病理学杂志

Purpose To explore the molecular features of diffuse large B-cell lymphoma(DLBCL)with high expression of MYC.Methods The clinical data of 45 cases of DLBCL were collected.Immunohistochemical EnVision method was used to classify the patients into the group with high expression of MYC and the group with low expression of MYC.All samples were subjected to DNA targeted sequencing and molecular typing was performed using the LymphGen online tool.Cellular origin was determined by using the Lymph2Cx method.The correlation be-tween MYC overexpression and clinicopathological parameters was analyzed by the x2 test and Fisher precise test.Survival curves were drawn and survival-related factors were analyzed u-sing Cox univariate and multivariate regression.ResultsCases were classified into DLBCL with high expression of MYC(n=17)and DLBCL with low expression of MYC(n=28).Com-pared to the group with low expression of MYC,the group with high expression of MYC had more PIM1,MYD88,CD79B,CD58 and PRDM1 mutations(76.5％vs 28.6％,70.6％vs 32.1％,58.8％vs28.6％,29.4％vs3.6％,29.4％vs 3.6％,P＜0.05),MCD were more frequently found(58.8％vs 10.7％,P=0.001),GCB were rarely found(17.6％vs 50.0％,P=0.030).Overall survival was significantly shorter in DLBCL with high expression of MYC(P＜0.05).Cox multi-factorial analysis showed that age was an independent prognostic factor for DLBCL(P＜0.05).Conclusion Patients with high expression of MYC were frequently characterized as MCD and ABC,and PIM1,MYD88,CD79B,CD58 and PRDM1 muta-tions were common.Patients with high expression of MYC had a poorer prognosis.

Primary tonsil squamous cell carcinoma:clinicopathological and whole exome sequencing analysis / 临床与实验病理学杂志

Purpose To explore the clinicopathological characteristics of tonsil squamous cell carcinoma(TSCC),and to explore the whole exome mutations and tumor mutational bur-den(TMB)in TSCC cases.Methods Ten patients with clini-cally and histopathologically confirmed TSCC and their clinico-pathological characteristics were collected,The expression of CK(AE1/AE3),CK5/6,p63,p40,p16 and Ki67 were meas-ured by two steps of EnVision,and the whole exome sequencing(WES)and TMB were conducted in 3 of them.Results A-mong the 10 patients,there were 6 females and 4 males which aged from 43 to 76 years old.Microscopically,the cancer cells infiltrated into the subdermis of crypts in the form ofnests and irregular cord,accompanied by comedo like necrosis,intercellu-lar bridges,and varying degrees of keratinization.Obvious atyp-ia and mitotic figures were easily seen.Follow-up was available in all cases,ranging from 6 to 45 months.Nine cases had sur-vived.Immunohistochemistry staining showed that all cases were positive for CK(AE1/AE3),CK5/6,p63,p40,and p16 was positively expressed in three cases,and the proliferation index Ki67 ranged from 40％to 90％.The WES of three cases showed that ARID1B and LRP6 were common cancer susceptibility genes,and WDFY4,ZFHX4 exhibited higher mutation rates,which were both 3/3.The TMB analysis showed that one out of three cases was＞9 mut/Mb.Conclusion The early symptoms of TSCC are not obvious that lead to easily missed and misdiag-nosed.The WES analyses suggest that WDFY4 and ZFHX4 had a higher mutation rate.The TMB analysis suggests that some TSCC patients may benefit from immunotherapy.