Epidermoid cysts of the orofacial region: A clinico-pathological study of 13 cases with review of literature

Background: Epidermoid cysts (ECs) are uncommon benign cystic lesions derived from the germinative epithelium. Head and neck ECs constitute only 7% of all ECs whereas only 1.6% are seen intraorally. The floor of the mouth is the commonest intraoral site whereas tongue, lips, buccal mucosa, and jaws are less commonly involved intraoral sites. To date, very few large case series of ECs of head and neck have been published. To the best of our knowledge, this is the third-largest case series of 11 intraoral ECs along with 2 extra-oral cases in the pre-auricular region. Aims: To highlight the typical and atypical features of ECs in the common as well as rare sites and draw attention to its consideration as a differential diagnosis for head and neck masses. Settings and Design: Archival data of 13 histopathological cases identified as ECs were analyzed from the Department of Oral Pathology at a tertiary dental hospital and college in New Delhi from 2007 to 2020. Materials and Methods: The demographic, clinical, radiographic, histopathological features, and treatment modalities were recorded and analyzed. Statistical Analysis Used: Appropriate statistical tests were used. Results: The study found strong male predilection in the ratio of 10:3 with an average age of presentation as 28 years. The pre-auricular region and floor of the mouth were the common sites involved followed by buccal mucosa, lips, and jaws. All patients presented with slowly growing swelling with dysphagia, dyspnea, and dysphonia seen in larger cysts on the floor of the mouth. Microscopically, all cases were lined with stratified squamous epithelium filled with laminated layers of keratin. Two cases showed the presence of melanin. One case showed recurrence even after complete surgical excision. Conclusion: ECs, though a rare entity, should be considered in differential diagnosis for head and neck masses and require close follow-up due to their potential for malignant transformation.

Inmunolocalización de citoqueratina 14, 19 y Ki-67 en el epitelio de pacientes con agrandamiento gingival

R E S U M E N En pacientes con ortodoncia aparecen eventos patológicos no deseados como agrandamiento gingival inducido por tratamiento de ortodoncia (AGTO) o hipertrofia gingival. El objetivo del estudio es identificar la distribución inmunohistoquímica de citoqueratina CK-14, CK-19 y Ki-67 en epitelio gingival de pacientes con AGTO. Se seleccionaron I3 pacientes divididos en: grupo control (n=6), conformado por individuos periodontalmente sanos no portadores de aparatología ortodóntica y grupo test (n=7), integrado por pacientes con AGTO. Los marcadores CK-14, CK-19 y Ki-67 fueron identificados mediante inmunohistoquímica con anticuerpos monoclonales y observados en un microscopio óptico Leica DM 500. En los pacientes del grupo test el tejido epitelial se mostró hipertrófico con pérdida en la continuidad de la membrana basal. La CK-14 y CK-19 fue positiva en el epitelio de todos los sujetos evaluados, con una expresión positiva de alta intensidad en células de la lámina basal del grupo test. El promedio de células positivas para Ki-67 en el grupo test fue de 56%. En conclusión, la CK-14, CK-19 y Ki-67 son marcadores con elevada inmunoreactividad en tejido gingival de pacientes con AGTO portadores de ortodoncia.

During orthodontic treatment, unwanted pathological events such as gingival overgrowth induced by orthodontic treatment or gingival hypertrophy may appear The objective of this study is to identify immunohistochemical distribution of cytokeratin CK-14, CK-19 and Ki-67 in the gingival epithelium of patients with gingival overgrowth induced by orthodontic treatment. Thirteen patients were selected divided into: control group (n = 6), conformed of periodontally healthy individuals without orthodontic appliances and the test group (n = 7), conformed of patients with gingival overgrowth induced by orthodontic treatment. The biomarkers CK-14, CK-19 and Ki-67 were identified by immunohistochemistry with monoclonal antibodies and observed in a Leica DM 500 optical microscope. Hypertrophic epithelial tissue with loss of continuity of the basement membrane was found in the test group patients. CK-14 and CK-19 were positive in the epithelial tissue of all the subjects evaluated, with a high intensity positive expression in the cells of the basal lamina of the test group. The average number of cells positive for Ki-67 in test group was 56%. In conclusion, CK-14, CK-19 and Ki-67 are biomarkers with high immunoreactivity in the gingival tissue of patients with gingival overgrowth induced by orthodontic treatment.

Durante o tratamento ortodôntico, eventos patológicos indesejados como o crescimento gengival induzido pelo tratamento ortodôntico (CGTO) ou hipertrofia gengival podem aparecer: O objetivo deste estudo é identificar a distribuição imuno-histoquímica das citoqueratinas CK -14, CK-19 e Ki-67 no epitélio gengival de pacientes com CGTO. Foram selecionados 13 pacientes divididos em: grupo controle (n=6), conformado por indivíduos periodontalmente saudáveis sem aparelhos ortodônticos e o grupo teste (n=7), conformado por pacientes com CGTO. Os biomarcadores CK-14, CK-19 e Ki-67 foram identificados por imuno-histoquímica com anticorpos monoclonais e observados em microscópio óptico Leica DM 500. Tecido epitelial hipertrófico com perda de continuidade da membrana basal foi encontrado nos pacientes do grupo teste. CK-14 e CK-19 foram positivos no tecido epitelial de todos os sujeitos avaliados, com expressão positiva de alta intensidade nas células da lâmina basal do grupo teste. O número médio de células positivas para Ki-67 no grupo teste foi de 56%. Em conclusão, CK-14, CK-19 e Ki-67 são biomarcadores com alta imunorreatividade no tecido gengival de pacientes com CGTO.

Study on the correlation of cyfra21-1 and modified ultrasound in connective tissue disease associated with interstitial lung disease / 中华风湿病学杂志

Objective:To investigate the correlation between cytokeratin 19 fragment(CYFRA21-1), modified ultrasound B-line and connective tissue disease associated with interstitial lung disease (CTD-ILD).Methods:The data of 112 patients with CTD hospitalized in the Department of Rheumatology and Immunology of the Second Hospital of Fujian Medical University from September 2019 to December 2021 were retrospectively collected. Sixty patients in the CTD-ILD group and 52 patients in the connective tissue disease without interstitial lung disease (CTD-noILD) group were included. The t-test and χ2 test were used to compare the demographic characteristics and tumor-associated antigens of the two groups of patients. Modified ultrasound score and HRCT Warrick score were evaluated by Pearson correlation analysis. In addition, the relationship between CYFRA21-1, modified ultrasound score and Warrick score were evaluated, and the diagnostic efficacy of CYFRA21-1 and modified ultrasound of CTD-ILD was evaluated and analyzed by binary logistic regression analysis. Results:Patients in the CTD-ILD group had higher CYFRA21-1 concentrations than the CTD-no-ILD group[5.74(4.25, 9.79) ng/ml vs. 2.79(2.21, 3.23) ng/ml, Z=45.94, P<0.001], patients in the CTD-ILD group had higher modified ultrasound scores than the CTD-no-ILD group [44.5(36.5, 60.0) vs. 5.0 (3.2, 6.8), P<0.001]. Modified ultrasound score was positively correlated with Warrick score ( r=0.93, P<0.001) and CYRFA21-1 was positively correlated with modified ultrasound score ( r=0.39, P=0.042). The sensitivity of CYFRA21-1 in determining CTD-ILD was 81.7% and the specificity was 92.3% [ AUC (95% CI)=0.88(0.81, 0.95), P<0.001], the sensitivity of modified ultrasound B-line to determine CTD-ILD was 96.4% and the specificity was 92.9% [ AUC (95% CI)=0.99 (0.97, 1.00), P<0.001]. History of smoking[ OR(95% CI)=9.26(1.11, 77.12), P=0.040] and elevated CYFRA21-1 concentration[ OR(95% CI)=19.40(4.89, 76.95), P<0.001] were risk factors for CTD-ILD. Conclusion:CYFRA21-1 is expected to be a serum marker indicating concomitant ILD in patients with CTD. Modified ultrasound B-line to determine concomitant ILD in CTD patients has good diagnostic utility and can reflect the severity of pulmonary fibrosis in CTD-ILD patients.

The value of enhanced CT combined with serum CYFER21-1 and NSE in the diagnosis of lymph node metastasis in patients with NSCLC / 中国医师杂志

Objective:To investigate the value of preoperative enhanced CT combined with serum cytokeratin fragment 19 (CYFER21-1) and neuron-specific enolase (NSE) in the diagnosis of lymph node metastasis in patients with non-small cell lung cancer (NSCLC).Methods:160 patients with NSCLC admitted to Linyi Cancer Hospital from October 2018 to October 2021 were retrospectively selected. All patients received surgical treatment in our hospital, and 84 patients with lymph node metastasis (metastatic group) and 76 patients without lymph node metastasis (non-metastatic group) were confirmed after surgery. The features of enhanced CT images and serum CYFER21-1 and NSE levels were compared between the two groups before operation, and the value of each index in the diagnosis of lymph node metastasis in patients with NSCLC alone and in combination was analyzed by receiver operating characteristic (ROC) curve.Results:The proportions of patients with lesion diameter ≥3.0 cm, pleural depression, lymph node enlargement shown by CT, lymph node short diameter ≥10 mm, lymph node boundary ambiguity and lymph node enhancement in metastatic group were significantly higher than those in non-metastatic group, with statistical significance (all P<0.05). Serum CYFER21-1 and NSE levels in metastatic group were significantly higher than those in non-metastatic group, with statistical significance (all P<0.05). The area under curve (AUC) of CYFER21-1 and NSE levels in the diagnosis of lymph node metastasis in NSCLC patients were 0.652 and 0.845, respectively, and the diagnostic cut-off values were 4.81 ng/ml and 24.14 ng/ml, respectively. The sensitivity and specificity of CYFER21-1+ NSE+ enhanced CT in the diagnosis of lymph node metastasis in NSCLC patients were 91.67% and 94.74%. Conclusions:Preoperative enhanced CT is of certain clinical value in the diagnosis of lymph node metastasis in NSCLC patients. Combined with serum CYFER21-1 and NSE levels, enhanced CT can further improve the sensitivity and specificity of diagnosis.

Increased Hes1 expression to regulate differentiation of liver epithelial progenitor cells into cholangiocytes / 西安交通大学学报(医学版)

【Objective】 To use hairy enhancer of split 1 (Hes1) to regulate the differentiation of liver epithelial progenitor cells (LEPCs) into cholangiocytes. 【Methods】 The vectors, pTet-on and pTRE2hyg-Hes1, were transfected into LEPCs. The expression of Hes1 was induced by doxycycline (DOX) with different concentrations (0, 0.1, 1, 5, 10, 50, 100 and 500 μg/mL). The expressions of Hes1, molecular markers of hepatocyte and cholangiocyte, glutathione synthetase (Gss), keratin 19 (Krt19) and hepatic nuclear factor 1β (HNF1β) in LEPCs were verified by Western blotting, RT-PCR, Real-time PCR, immunocytochemistry and immunofluorescence. 【Results】 The expression of Hes1 in LEPCs transfected by pTet-on/pTRE2hyg-Hes1 was increased by 11.21 fold when induced by DOX at 10 ug/mL, which drove the LEPCs to differentiate into biliary epithelial cells. With increasing expression of Hes1, cholangiocyte markers, Krt19 and HNF1β, were significantly upregulated, while the hepatocyte marker, Gss, was obviously downregulated. 【Conclusion】 DOX at 10 μg/mL may induce a suitably up-regulated expression of Hes1 in LEPCs double-transfected by pTet-on and pTRE2hyg-Hes1, and the suitable high-expression rather than over-expression of Hes1 can regulate LEPCs to differentiate into cholangiocytes.

Middle phalanx epidermoid cyst, an unusual site: A case report

Background: Epidermoid cysts are cutaneous cysts frequently observed among farmers, factory workers, carpenters, and tailors due to their occupations’ predilection for repetitive minor hand trauma. Case Presentation: A 66-year-old male laborer presented to our hospital with a swelling on the palmar aspect of his right middle finger. The swelling was accompanied by pain and an inability to work over the previous four months. A 2.5 cm × 2.5 cm smooth, tender, immobile, non-compressible hard swelling was detected just above the middle phalanx during the clinical examination on the volar aspect of the right middle finger. The diagnosis was confirmed by histopathology after excision. The patient’s range of motion improved significantly following surgery, and his pain subsided. Conclusion: We present a novel location for an epidermal inclusion cyst and the importance of histopathologic evaluation in this case. Additionally, we emphasize the significance of extensively checking the cyst’s epithelium to guarantee complete capsule removal and rule out any additional pathology.

Expression and regulation of keratin 6B in CD44+ bladder cancer stem cells / 中国组织工程研究

BACKGROUND: Bladder cancer stem cells could promote the recurrence and drug resistance of bladder cancer. Numerous studies have shown that keratin 6B (KRT6B) is involved in the production and progression of tumors, and is closely related to the prognosis of tumors. OBJECTIVE: To observe the expression of keratin 6B in CD44+ bladder cancer stem cells and to show the influence of keratin 6B on proliferation, migration, and self-renewal of bladder cancer stem cells, and to further explore the effect of keratin 6B expression on the prognosis of bladder cancer patients. METHODS: (1) CD44+ 5637 bladder cancer stem cells were isolated by magnetic active cell sorting. Cancer stem cell-related gene expression of SOX2, OCT4, and NANOG was detected via real-time polymerase chain reaction. The spheroid formation assay was used to detect the ability of self-renewal of cancer stem cells in CD44+ cells. Keratin 6B expression was detected in CD44+ bladder cancer stem cells by real-time polymerase chain reaction. (2) The CD44+5637 bladder cancer stem cells were divided into two groups. In the keratin 6B siRNA group, keratin 6B small interfering RNA was transfected into CD44+ bladder cancer stem cells. Untransfected CD44+ bladder cancer stem cells were used as the black control group. Cells were collected at 2 days post-transfection. The proliferation, migration, and self-renewal capacity of keratin 6B siRNA CD44+ bladder cancer stem cells were detected by the colony and wound healing assay and spheroid formation respectively. (3) Totally 24 bladder cancer tissues were used by immunohistochemistry to analyze the expression of CD44v6 and keratin 6B. (4) ONCOMINE database was used to analyze the effect of keratin 6B expression on the overall survival of bladder cancer. RESULTS AND CONCLUSION: (1) Cancer stem cell-related genes (SOX2, OCT4, NANOG) and keratin 6B expression was higher in CD44+ cells isolated by magnetic active cell sorting compared with CD44- cells (P < 0.05). Cell proliferation, migration, and in vitro spheroid formation were significantly increased (P < 0.05). Keratin 6B small interfering RNA down-regulated the expression of keratin 6B in CD44+ bladder cancer stem cells (P < 0.05). (2) Compared with the blank control group, the proliferation and migration of CD44+ bladder cancer stem cells after transfection of keratin 6B small interfering RNA (P < 0.05), and the number of tumorsphere significantly diminished (P < 0.05); the expression of Notch1 and Hes1 mRNA increased (P < 0.05). (3) Keratin 6B and CD44v6 were significantly different in bladder cancer tissue (P=0.006). The overall survival rate of bladder cancer patients with high expression of keratin 6B was lower than that of patients with low expression of keratin 6B. (4) The results showed that keratin 6B was highly expressed in CD44+ bladder cancer stem cells, and could promote the proliferation, migration, and self-renewal capacity of bladder cancer stem cells. The high expression of keratin 6B contributes to improving the survival of bladder cancer patients.

Effect of HaCaT cells with CRISPR-Cas9-induced KRT5 mutation on co-cultured human melanocytes / 中华皮肤科杂志

Objective:To investigate the effect of KRT5 knockdown in keratinocytes on melanin content in co-cultured melanocytes, and to explain mechanisms underlying formation of hyperpigmented lesions in reticulate pigmented anomaly of the flexures (Dowling-Degos disease, DDD) .Methods:HaCaT cells with heterozygous mutations in the KRT5 gene were obtained by using clustered regularly interspaced short palindromic repeats (CRISPR) -CRISPR-associated protein 9 (Cas9) technology (experimental group) , and HaCaT cells transfected with non-targeting single guide RNA:Cas9 protein complex served as control group, both of which were in vitro co-cultured with primary human melanocyte cells (HEMn) separately. Immunofluorescence study was conducted to determine the expression of cytokeratin and melanosomes in co-cultured cells; melanin content was detected in melanocytes in different co-culture groups, which were obtained by differential trypsinization. Immunohistochemical study was performed to determine the expression of melanocyte-specific premelanosome protein 17 (Pmel17) in skin lesions in a patient with DDD carrying a KRT5 mutation and normal skin tissues in a healthy control. Results:Sanger sequencing showed a heterozygous mutation (c.1delA) at the initiation codon of exon 1 of the KRT5 gene in HaCaT cells in the experimental group, but no mutation in the KRT5 gene in the control group. Western blot analysis showed that the KRT5 protein expression was significantly lower in the experimental group (0.60 ± 0.05) than in the control group (1.00 ± 0.00, t = 32.38, P = 0.001) . Compared with the co-culture system in the control group, the number of Pmel17-labeled melanosomes markedly increased with the melanin content elevated by 52.5% ( t = -3.48, P = 0.025) in the HEMn cells co-cultured with HaCaT cells in the experimental group. Immunohistochemical study showed that the Pmel17 expression increased in the skin lesions in the DDD patient with KRT5 mutation compared with the normal skin tissues in the healthy control. Conclusion:The effect of HaCaT cells with CRISPR-Cas9-induced KRT5 mutation on the co-cultured HEMn melanocytes was verified by the successfully established in vitro co-culture system, which provides a primary cell model for further studies on interaction mechanisms between keratinocytes and melanocytes, and on pathogenesis of skin pigmentation abnormalities.

Value of different tumor markers for the diagnosis and prognosis prediction of colon cancer in patients / 中国基层医药

Objective:To comprehensively analyze the clinical values of serum tumor markers of colon cancer, including carbohydrate antigen 724 (CA724), cytokeratin 19 fragment antigen (CYFRA21-1), carbohydrate antigen199 (CA199) and carcinoembryonic antigen (CEA) in the diagnosis and prognosis prediction of colon cancer in patients.Methods:The clinical data of 160 patients with colon cancer who received treatment in Zhuji Central Hospital from January 2018 to December 2020 (observation group) and the clinical data of 156 patients with benign colon polyps who concurrently received physical examination (control group) were retrospectively analyzed. All patients underwent CA724, CYFRA21-1, CA199 and CEA tumor marker screening. The levels of tumor markers, the positive rate of a single tumor marker, and the positive rate of a combination of four markers were compared between the control and observation groups. The levels of tumor markers were compared among different pathological stages. The levels of serum tumor markers were compared among patients with different prognoses based on 1-year follow-up data.Results:CA199-positve rate, CEA-positive rate, CYFRA21-1-positve rate, CA724-positive rate, and the positive rate of a combination of four tumor markers were 85.63% (137/160), 86.88% (139/160), 71.88% (115/160), 85.00% (136/160), and 95.63%(153/160), respectively, which were significantly higher than those in the control group ( χ2 = 8.64, 10.28, 8.33, 9.93, 7.27, all P < 0.001). Serum CA199, CEA, CYFRA21-1 and CA724 levels in patients with stage III-IV colon cancer were (58.96 ± 13.59) U/mL, (38.69 ± 11.84) μg/L, (14.78 ± 3.68) μg/L, (23.68 ± 5.38) U/mL, respectively, which were significantly higher than those in patients with stage I-II colon cancer [(48.35 ± 9.03) U/mL, (23.96 ± 12.25) μg/L, (9.57 ± 2.53) μg/L, (13.02 ± 4.32) U/mL, t = 10.29, 12.02, 8.47, 10.54, all P < 0.001). One-year follow-up results showed that serum levels of CA199, CEA, CYFRA21-1, CA724 in patients with recurrence and metastasis of colon cancer were (38.68 ± 3.04) U/mL, (17.12 ± 4.96) μg/L, (8.94 ± 2.32) μg/L, (11.22 ± 1.94) U/mL, which were significantly higher than those in patients without recurrence of colon cancer [(30.02 ± 2.95) U/mL, (3.75 ± 1.06) μg/L, (3.06 ± 1.15) μg/L, (6.28 ± 1.53) U/mL, t = 8.73, 11.02, 7.72, 7.57, all P < 0.001]. Conclusion:Serum levels of CEA, CA199, CA724 and CYFRA21-1 can be used as important indicators for diagnosis and prognosis prediction of colon cancer.

Research Advances in the Role of Keratins in Gastrointestinal Cancer / 中国医学科学杂志(英文版)

The incidence and mortality rates of gastrointestinal (GI) cancer remain high. Despite constant improvements in diagnostic and therapeutic techniques, the early diagnosis, mid- and late-stage treatment, drug tolerance, and cancer recurrence and metastasis in GI cancer remain challenging. In this review article we summarize the recent research advance in the roles of keratins in GI cancer, with the hope that they will become efficient biomarkers for the prediction, diagnosis, or treatment of these malignancies.

The effect of androgen on wool follicles and keratin production in Hetian sheep / Efeito do androgênio na produção de folículos e queratina em ovinos Hetianos

Abstract To investigate the optimal androgen concentration for culturing Hetian sheep wool follicle and to detect effects of androgen concentration on wool follicle cell proliferation and apoptosis using immunofluorescence labeling and real-time quantitative fluorescence determinations of wool keratin-associated protein gene expression levels. Wool follicles were isolated by microdissection and wool follicles and skin pieces were cultured in various concentrations of dihydrotestosterone (DHT) in culture medium. Next, daily lengthwise growth measurements of wool follicles were obtained using a microscopic micrometer. Cultured Hetian wool follicles were stained using the SACPIC method to reveal wool follicle structure, while sheep skin slices were used to observe cell proliferation by immunostaining and cell apoptosis using the TUNEL method. At the molecular biological level, keratin-associated protein (Kap) gene expression was studied using wool follicles cultured for various numbers of days in vitro. Effects of androgen concentrations on Hetian wool follicle growth and development were experimentally studied. EdU proliferation assays revealed that androgen promoted cell proliferation within wool follicle dermal papillae. TUNEL apoptosis detection demonstrated that androgen treatment could delay cell apoptosis. Quantitative reverse transcription polymerase chain reaction (qPCR) results demonstrated that gene expression level patterns of Hetian mountain sheep super-high sulfur protein. Kap1.1, KIF1.2, Kap2.12 and Kap4.2 gene expression level of the mountainous experimental group was significantly higher than plains Hetian sheep. An androgen concentration of 100 nM can promote the growth of Hetian wool follicle cells in vitro, resulting in overexpression of some genes of the Kap family.

Resumo Investigar a concentração ideal de andrógenos em cultura de folículos pilosos de carneiro Hetiano e detectar os efeitos da concentração de andrógenos na proliferação e apoptose de células foliculares, por meio de imunofluorescência e de determinação quantitativa, em tempo real, da fluorescência dos níveis de expressão gênica de proteína associada à queratina. Folículos pilosos foram isolados por microdissecção, e folículos de lã e pedaços de pele foram cultivados em várias concentrações de di-hidrotestosterona (DHT) em meio de cultura. Em seguida, medições diárias de crescimento longitudinal dos folículos capilares foram obtidas usando um micrômetro microscópico. Folículos de lã cultivados de Hetianos foram corados pelo método SACPIC para revelar a estrutura do folículo piloso, enquanto fatias de pele de carneiro foram usadas para observar a proliferação celular por imunocoloração e apoptose celular por meio do método TUNEL. Em âmbito da biologia molecular, a expressão gênica da proteína associada à queratina (Kap) foi estudada usando folículos capilares cultivados por vários dias, in vitro. Os efeitos das concentrações de andrógenos no crescimento e desenvolvimento dos folículos de lã de Hetianos foram estudados experimentalmente. Ensaios de proliferação de EdU revelaram que o andrógeno promoveu a proliferação celular dentro das papilas dérmicas do folículo piloso. A detecção de apoptose por TUNEL demonstrou que o tratamento com andrógeno poderia atrasar a apoptose celular. Os resultados da reação em cadeia da polimerase transcrição reversa quantitativa (qPCR) demonstraram que os padrões de expressão gênica da proteína de enxofre Kap1.1, KIF1.2, Kap2.12 e Kap4.2 foram significativamente maiores no grupo de ovinos Hetianos de montanha. Uma concentração de androgênio de 100 nM pode promover o crescimento de células foliculares de lã de Hetianos in vitro, resultando na superexpressão de alguns genes da família Kap.

Application value of spiral CT combined with a disintegrin and metalloproteinase 8, neuron-specific enolase and cytokeratin 19 fragment measurements in the clinical diagnosis of lung cancer / 中国基层医药

Objective:To investigate the application value of spiral CT combined with a disintegrin and metalloproteinase 8 (ADAM8), neuron-specific enolase (NSE) and cytokeratin 19 fragment (CYFRA21-1) measurements in the clinical diagnosis of lung cancer.Methods:Fifty patients with lung cancer who received treatment in Shanxi Rongjun Hospital from February 2018 to December 2019 were included in the lung cancer group. Fifty patients with benign lung disease who concurrently received treatment in Shanxi Rongjun Hospital were included in the benign lung disease group. Fifty 50 healthy controls who concurrently received treatment in Shanxi Rongjun Hospital were included in the control group. Serum levels of ADAM8, NSE and CYFRA211 were compared among the three groups. Three groups received spiral CT scans. ADAM8-, NSE- and CYFRA211-positive detection rates were analyzed. The receiver operating characteristic (ROC) curve was used to analyze the efficacy of different methods in the diagnosis of lung cancer. Serum ADAM8, NSE and CYFRA211 levels in patients with lung cancer at different TNM stages were compared.Results:Serum levels of ADAM8, NSE and CYFRA211 in the lung cancer and benign lung disease groups were (381.69 ± 34.82) ng/L, (255.28 ± 30.48) ng/L, (16.87 ± 3.11) μg/L, (9.27 ± 2.11) μg/L,(13.54 ± 8.10) μg/L, (3.01 ± 1.34) μg/L, respectively, which were significantly higher than those in the control group [(225.83 ± 24.19) ng/L, (7.42 ± 2.35) μg/L, (1.78 ± 1.02) μg/L, t = 5.352, 25.994, 4.142, 17.142, 5.165, 10.186, all P < 0.05]. Serum levels of ADAM8, NSE and CYFRA211 in the lung cancer group were significantly higher than those in the benign lung disease group ( t = 19.316, 14.299, 9.069, all P < 0.05). The positive detection rate of lung cancer by spiral CT combined with ADAM8, NSE and CYFRA211 measurements was 92.00%, which was significantly higher than that by spiral CT combined with a single detection ( χ2 = 7.862, 9.000, 11.422, 9.000, all P < 0.05). The ROC curve analysis revealed that the area under the ROC curve, sensitivity and specificity of the combined method were 0.916, 0.920 and 0.900, respectively, which were significantly higher than those of spiral CT combined with a single detection. Serum ADAM8, NSE and CYFRA211 levels at TNM stages III-IV were (430.18 ± 36.43) ng/L, (20.05 ± 3.49) μg/L, (15.93 ± 8.22) μg/L, respectively, which were significantly higher than those at TNM stages I-II [(314.72 ± 30.85) ng/L, (12.49 ± 2.67) μg/L, (10.25 ± 6.35) μg/L, t = 11.777, 8.312, 2.644, all P < 0.05). Conclusion:Spiral CT combined with serum ADAM8, NSE and CYFRA211 levels can greatly increase the diagnostic sensitivity and specificity of lung cancer, which is worthy of clinical application.

Changes and clinical significance of serum matrix metalloproteinase-9, squamous cell carcinoma antigen, cytokeratin-19 fragment, carcinoembryonic antigen and neuron-specific enolase levels in peripheral lung cancer / 中国基层医药

Objective:To investigate the changes and clinical significance of serum matrix metalloproteinase-9 (MMP-9), squamous cell carcinoma antigen (SCC), cytokeratin 19 fragment (CYFRA21-1), carcinoembryonic antigen (CEA) and neuron-specific enolase (NSE) in peripheral lung cancer.Methods:Sixty-eight patients with peripheral lung cancer who received treatment in Luqiao Hospital of Taizhou Enze Medical Center (Group) between January 2017 and January 2020 were included in the observation group. Sixty-five patients with benign lung diseases who concurrently received treatment in the same hospital were included in the observation group 1, and another 65 healthy participants who concurrently received physical examination were included in the control group. Serum levels of MMP-9, CYFRA21-1, SCC, NSE and CEA were compared among the three groups. The sensitivity and specificity of using these indicators alone and in combination in the diagnosis of peripheral lung cancer were compared.Results:Serum levels of MMP-9, CYFRA21-1, SCC, NSE and CEA in the observation group (14.98 ± 2.10) ng/mL, (17.13 ± 2.71) ng/mL, (1.98 ± 0.41) μg/mL, (24.13 ± 2.10) ng/mL and (17.10 ± 2.10) ng/mL, respectively, which were significantly higher than those in the observation group 1 [(9.12 ± 1.41) ng/mL, (10.12 ± 1.58) ng/mL, (1.37 ± 0.31) μg/mL, (16.31 ± 1.78) ng/mL, (12.13 ± 1.79) ng/mL] and control group [(5.10 1 ± 0.68) ng/mL, (6.02 ± 0.94) ng/mL, (0.71 ± 0.11) μg/mL, (11.10 ± 1.02) ng/mL, (8.13 ± 1.02) ng/mL] ( F1 = 932.781, F2 = 737.100, F3 = 368.591, F4 = 989.851, F5 = 462.291, all P < 0.05). Serum levels of MMP-9, CYFRA21-1, SCC, NSE and CEA in patients with stage I-II peripheral lung cancer were (11.12 ± 2.10) ng/mL, (9.12 ± 1.85) ng/mL, (1.52 ± 0.21) μg/mL, (18.12 ± 3.02) ng/mL, (7.52 ± 1.02) ng/mL, respectively, which were significantly lower than those in patients with stage III-IV peripheral lung cancer [(15. 89 ± 2.18) ng/mL, (21.56 ± 2.11) ng/mL, (2.04 ± 0.31) μg/mL, (28.15 ± 2.62) ng/mL, (15.12 ± 1.55) ng/mL, t1 = 9.013, t2 = 25.146, t3 = 7.714, t4 = 14.586, t5 = 22.705, all P < 0.05]. The sensitivity (83.33%) and specificity (86.67%) of combined detection of all indicators were significantly higher than those of single detection of MMP-9 (50.00%, 59.68%), CEA (50.00%, 61.29%), CYFRA21-1 (66.67%, 58.06%), SCC (50.00%, 54.84%) or NSE (66.67%, 58.06%) (all P < 0.05). Conclusion:Serum levels of MMP-9, CYFRA21-1, SCC, NSE and CEA in patients with peripheral lung cancer are significantly increased, which has an important value in the diagnosis of peripheral lung cancer. The combined detection of the above indicators can increase the diagnostic accuracy of peripheral lung cancer in the clinic.

Effect of hot spring bathing on rheumatoid related indicators and joint pain / 上海预防医学

Objective To observe the intervention effect of typical hot spring bathing in Guizhou province on joint pain, serum anti-keratin antibody(AKA), anti-perinuclear factor antibody(APF)and anti-cyclic citrullinated peptide antibody(CCP). Methods A total of 160 people with joint pain symptoms from five typical hot spring areas in Guizhou province were selected as the subjects. They were treated with hot spring bathing intervention for 4 weeks, once a day, 5 times a week, 40 to 50 minutes each time. According to the evaluation index of physiotherapy natural mineral water in the Code for Geological Exploration and Evaluation of Natural Warm Mineral Water Resources(GB/T 13727-2016)and geological types, the five typical hot springs were divided into three different types, namely water temperature type hot springs(water temperature > 36 ℃), metasilicate type hot springs(metasilicate > 50 mg/L)and warm mineral spring type hot springs(total dissolved solids > 1 000 mg/L). WHO pain grading standard was used to score the degree of joint pain before and after hot spring bathing intervention. Serum APF, AKA and CCP antibodies were detected by ELISA kit before and after hot spring bathing. Results The joint pain score of the subjects was 2.60±0.60, and the joint pain score of the total population decreased after intervention(0.61±0.57, P < 0.05). Before intervention, the joint pain scores of water temperature type, metasilicic acid type and warm mineral spring type were 2.78±0.96, 1.98±1.15 and 3.31±0.57, respectively. After intervention, the scores of joint pain of the three kinds of hot spring bathing patients all decreased(P < 0.05), and were 0.50±0.65, 0.48±0.74 and 0.85±0.90, respectively. Before intervention, AKA(ng/L)and CCP(μg/mL)antibody levels of the observed subjects were 34.89±16.06 and 107.58±10.40, respectively, which significantly decreased after intervention(both P < 0.05), namely 26.06±10.68 and 102.93±6.01, respectively. AKA(ng/L)was 35.04±20.01 before intervention, but decreased significantly after intervention(26.61±7.54, P < 0.05). AKA(ng/L)and CCP(μg/mL)were 31.09±17.26 and 106.51±10.13 before intervention, respectively. After intervention, the above two antibody indexes significantly decreased(all P < 0.05)to 24.53±13.98 and 98.57±5.68, respectively. Before intervention, the AKA(ng/L), APF(ng/mL)and CCP(μg/mL)antibody levels were 38.40±8.66, 349.46±118.43 and 104.96±9.66, respectively. After intervention, the above three antibody indexes significantly decreased(all P < 0.05). The values were 34.00±7.55, 269.38±127.55 and 101.65±3.04, respectively. Conclusion The typical hot spring bathing intervention in Guizhou province can relieve the symptoms of joint pain, and the three types of hot springs can reduce the levels of AKA, APF and CCP antibodies to different degrees, and the warm mineral spring type of hot spring is better than the other types of hot spring.

Effect of high temperature sand fried processing on protein in Manis Squama based on "Proteomics-Modifications" methods / 中草药

Objective: To study the effect of high temperature sand fried processing on the chemical constituents in Manis Squama based on the changes of proteins, peptides, and modifications. Methods: Nano LC-MS/MS was used to analyze and identify proteins and peptides in Manis Squama before and after processing. The PTMs including deamidation and oxidation occurred in Manis Squama during processing were also investigated. Results: The results showed that Manis Squama consisted of keratins, connexin, desmoplakin, and some isomerases, which can promote the keratinous structure formation. After processing, the number of identified proteins and peptides in soluble fraction did not show significant change, while the number of proteins and peptides in in-soluble faction were decreased significantly. The number of deamidation was increased significantly, and the number of deamidation on Asn and Gln in Keratins was increased significantly. There were no significantly change on molecular weight distribution and GRAVY value. Conclusion: High temperature sand fried processing can decrease the protein and peptide identifications of Manis Squama, but significantly increase the identifications from Keratins and other structural proteins, and significantly increase the number of deamidation, which might help to increase the releasing and dissolution of soluble proteins and peptides. In this study, we provided important evidences on revealing the effect of processing on the chemical constituents changing in Manis Squama, also provided ideas and methods for searching and evaluation of Manis Squama alternative resources.

Analysis of the green turtle esophagus Chelonia mydas (Linnaeus, 1758), Testudines, Cheloniidae / Análisis del esófago de la tortuga verde Chelonia mydas (Linnaeus, 1758), Testudines, Cheloniidae

The esophagus is a muscular tube whose function is to transport food from the mouth to the stomach and basically contain the same layers as the rest of the digestive tract. The morphological knowledge of structures involved in basic physiological processes in organisms is extremely important in the acquisition of ecological and evolutionary knowledge about species. This study produces morphological information proving the structures found in the green turtle's esophagus. The animals were dissected according to the methodology used by Wyneken, allowing analysis of external and internal morphological characteristics of the esophagus. The samples were processed for microscopic analysis by light and scanning electron microscopy. It was observed that the esophagus presents thin and conical dermal papillae facing the stomach throughout the mucosa to the gastroesophageal portion, decreasing its size at the end of its extension, covered by a thick keratin layer on keratinized pavement epithelial tissue. It is concluded that the esophagus of Chelonia mydas is covered by a layer of conical keratinized dermal papillae important to exert the mechanical and protective function of the mucosa of this organ, considering that the feeding of these animals is extremely abrasive, besides serving as protection so that the food does not return.

El esófago es un tubo muscular cuya función es transportar los alimentos desde la cavidad oral hasta el estómago y, básicamente, contienen las mismas capas que el resto del tracto digestivo. El conocimiento morfológico de las estructuras involucradas en los procesos fisiológicos básicos en los organismos es extremadamente importante en la adquisición de conocimientos ecológicos y evolutivos sobre las especies. Este estudio produce información morfológica que demuestra las estructuras encontradas en el esófago de la tortuga verde. Los animales se disecaron de acuerdo con la metodología utilizada por Wyneken, lo que permitió el análisis de las características morfológicas internas y externas del esófago. Las muestras se procesaron para análisis microscópico mediante microscopía electrónica de barrido y de luz. Se observó que el esófago presenta papilas dérmicas delgadas y cónicas que se enfrentan al estómago a través de la mucosa hasta la porción gastroesofágica, disminuyendo su tamaño al final de su extensión, cubierta por una capa gruesa de queratina sobre el tejido epitelial queratinizado. Se concluye que el esófago de Chelonia mydas está cubierto por una capa de papilas dérmicas queratinizadas cónicas importantes para ejercer la función mecánica y protectora de la mucosa de este órgano, considerando que la alimentación de estos animales es extremadamente abrasiva, además de servir de protección para que la comida no sea devuelta.

Fabrication Of Nanofibres By Electrospinning Using Keratin From Waste Chicken Feathers, Pva And Agnps

Objective: To prepare and characterise keratin from chicken feathers (CF), collected from the slaughter house, and to blend with poly vinly alcohol (PVA) and biosynthesised silver nanoparticles (AgNPs) and to convert into nanofibers by an elctrospinning process. Methods: The extraction of keratin from chicken feathers was done by sodium m-bisulphite. The solution was subjected to ammonium sulphate precipitation to separate keratin. The nanoparticles was synthesised using tridax procumbens. The isolated keratin and PVA was mixed in the ration 0f 50:50 with 1 ml of biosynthesised nanoparticles was blended and made into nanofibres by electrospinning technique. Results: The precipitated protein was analysed using FT-IR analysis confirming the presence of β-keratin in the sample isolated from chicken feathers and the concentration of keratin was estimated to be 1.85 g/ml. PVA solution with 4% w/v had the best film forming ability. The solution containing keratin, PVA and silver nanoparticles was prepared in various proportions. These solutions when subjected to electrospinning, fibrous network was observed in 50:50 (PVA: Keratin) ratio with 1 ml of synthesised silver nanoparticle solution. Hydrogen bonding between keratin and PVA indicated in the XRD analysis showed successful film forming of the nanofiber, the DSC analysis also showed similar results as the obtained peak was at 214 °C which is in between the characteristic heat degradation temperature of both the keratin and PVA. The thermogravimetric analysis (TGA) showed high thermal stability as the complete degradation of the nanofiber was observed at 420 °C. Incorporation of metal nanoparticles by herbal approach using tridax procumbens in the nanofibers provided the antimicrobial properties. The nanofibres obtained by electrospinning process appeared stable and continous for solutions containing no more than 50% wt of CF. The average diameter of the nanofibres increased as the CF content increased. Conclusion: Keratin isolated from the waste chicken feathers impregnated with biosyntheised silver nanoparticles using tridax procumbens and PVA can be converted into nanofibers by electrospinning process. Thus, the biocomposite nano fibers are shown as a novel eco-friendly material that must be adequately applied in the development of green composites for the biomedical applications such as wound dressings.

Prognostic significance of inflammation-based prognostic scoring in patients with upper urinary tract urothelial carcinoma

ABSTRACT Objectives: To investigate whether Glasgow Prognostic Score has prognostic significance in patients with upper urinary urothelial carcinoma. Patients and methods: We retrospectively reviewed the clinical records of 74 patients with upper urinary urothelial carcinoma. We set the cut-off value for C-reactive protein as 1.0mg/dL, and 3.5mg/dL for albumin as Glasgow Prognostic Score. Their blood data including albumin and C-reactive protein for Glasgow Prognostic Score and cytokeratin 19 fragment 21-1 as a tumor marker were measured before starting treatment. The patients were stratified into three groups with Glasgow Prognostic Score: The Group-1, albumin ≥3.5g/dL and C-reactive protein < 1.0mg/dL; Group-2, albumin < 3.5g/dL or C-reactive protein ≥1.0mg/dL; Group-3, albumin < 3.5g/dL and C-reactive protein ≥1.0mg/dL. Results: The median follow-up for all patients was 26.9 months (range: 10.9-91.1 months), during which 37 (50%) patients died. There was a significant difference in the estimated survival rate among the 3 groups stratified by Glasgow Prognostic Score. The estimated survival rate in the Group-1 was significantly higher than those in Groups 2 and 3. In the univariate analysis C-reactive protein, serum cytokeratin 19 fragment 21-1 and Glasgow Prognostic Score were significant predictors of overall survival. On the multivariate analysis, serum cytokeratin 19 fragment 21-1 and Glasgow Prognostic Score were independently associated with shorter overall survival. Conclusion: Our review suggests Glasgow Prognostic Score may play as a prognostic predictor for upper urinary urothelial carcinoma.

Screening and Characterisation of Keratin-Degrading Bacillus sp. from Feather Waste

Aim: This study focuses on the screening and characterisation of keratin-degrading Bacillus species from feather waste. Methods: Nine bacteria were isolated from feather waste obtained from a poultry layout at Egbeda local government secretariat, Ibadan, Nigeria. These bacteria were grown in basal medium with feather as primary source of carbon, nitrogen, sulfur and energy. Feather degrading bacteria were screened for both proteolytic activity and keratin degradation on skimmed milk agar and keratin azure medium respectively. They were also screened for their ability to degrade other keratin substrates such as hair and nail. Results: Three of the isolates with higher feather degradation levels also showed high proteolytic activity and release of azure dye. They were selected and identified phenotypically and genotypically using 16S rRNA sequencing as Bacillus licheniformis-K51, Bacillus subtilis-K50 and Bacillus sp.-K53. The bacteria were capable of degrading other keratin-containing substrates such as nail and hair. Bacillus subtilis-K50 and Bacillus licheniformis-K51 showed significant difference (P) in degradation among the three different keratin sources used yielding higher degradation with feather as keratin source with respective optical densities of 0.07 and 0.11 followed by hair and least in nails with optical densities of 0.05 and 0.07 respectively. Highest degradation of all the three keratin substrates was observed in Bacillus licheniformis-K51. Conclusion: The three isolated bacteria possess the ability to degrade keratin and utilize feather as keratin substrate. As a result, these can be considered as potential candidates for degradation and utilization of feather keratin.

Identificación bioinformática de Polimorfismos de Nucleótido Simple (PNSs) en genes candidatos para las características de la fibra en alpacas (Vicugna pacos) / Bioinformatic identification of Single Nucleotide Polymorphisms (SNPs) in candidate genes for fiber characteristics in alpacas (Vicugna pacos)

El objetivo fue identificar y predecir la ubicación de polimorfismos de nucleótido simple (PNSs) en genes relacionados al crecimiento de la fibra. Se realizó el estudio con un total de 31 genes de queratina (KRT9, KRT12, KRT13, KRT14, KRT16, KRT18, KRT20, KRT25, KRT1, KRT3, KRT5, KRT6a, KRT6b, KRT6c, KRT7, KRT8, KRT71, KRT80, KRT31, KRT32, KIRT40, KRT81, KRT82, KRT10, KRT15, KRT17, KRT19, KRT2, KRT4, KRT79 y KRT83) asociados con las características de lana, fibra y pelo en ovinos, cabras y humanos respectivamente, cuyas secuencias fueron encontradas en la base de datos del National Center for Biotechnology Information (NCBI). Mediante el uso de bases de datos y herramientas bioinformáticas como el Conserved Domains Database, Spling, y MegaBlast se logró ubicar secuencias únicas para cada gen. Estas secuencias fueron comparadas con los genomas de referencia Vicugna_pacos-2.0.2 y Vi_pacos_V1.0. Se identificaron 48 PNSs ubicados en las regiones intrónicas y exónicas de 22 genes. No se localizaron PNSs en o alrededor de los genes KRT10, KRT15, KRT17, KRT19, KRT2, KRT4, KRT6b, KRT6c y KRT79. El análisis comparativo entre las cuatro especies estudiadas permitió observar que los genes KRT81, KRT6b y KRT6c no están presentes en los genomas de referencia de alpaca, los genes KRT31, KRT14, KRT81, KRT83, KRT6b y KRT6c no están presentes en el genoma de referencia de ovino y los genes KRT31, KRT13, KRT81, KRT83, KRT6b y KRT6c no están presentes en el genoma de referencia de cabra.

The objective was to identify and predict the location of single nucleotide polymorphisms (SNPs) in genes related to fiber growth. The study was carried out with 31 keratin genes (KRT9, KRT12, KRT13, KRT14, KRT16, KRT18, KRT20, KRT25, KRT1, KRT3, KRT5, KRT6a, KRT6b, KRT6c, KRT7, KRT8, KRT71, KRT80, KRT31, KRT32, KIRT40, KRT81, KRT82, KRT10, KRT15, KRT17, KRT19, KRT2, KRT4, KRT79 y KRT83) associated with wool, fiber and hair characteristics in sheep, goat and human, respectively. These gene sequences were retrieved from the National Center for Biotechnology Information (NCBI) database. Using databases and bioinformatics tools such as the Conserved Domains database, Spling and Megablast, unique sequences for each gene were identified. These sequences were compared to the reference genomes: Vicugna_pacos-2.0.2 and Vi_pacos_V1.0 to identify single nucleotide polymorphisms (SNPs). In this manner, 48 SNPs were identified and localized in both intronic and exonic regions of 22 genes. We did not identify SNPs for KRT10, KRT15, KRT17, KRT19, KRT2, KRT4, KRT6b, KRT6c and KRT79. Comparative analysis among the four species studied allow to identify that sequences for KRT81, KRT6b and KRT6c genes are not present in the alpaca reference genomes. Similarly, genes KRT31, KRT14, KRT81, KRT83, KRT6b and KRT6c are not present in the ovine reference genome and, genes KRT31, KRT13, KRT81, KRT83, KRT6b and KRT6c are not present in the goat reference genome.