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To understand the current quality status and rearing situation of Bombyx Batryticatus, the authors collected 102 batches of Bombyx Batryticatus from different main producing areas and five major Chinese medicine markets from 2016 to 2018, and measured the properties and quality of the silk gland, to clarify the quality status of Bombyx Batryticatus from different producing areas and markets. In addition, 35 batches of Bombyx Batryticatus from 2019 to 2022 were used to verify the silk gland after revision. Moreover, Beauveria Bassiana was inoculated in the silkworm of 4-5 instars, and standardized rearing was carried out until they die. The death rate and the quality of Bombyx Batryticatus were measured to determine the differences in Bombyx Batryticatus of different instars, and explore the rationality of the infection age of Bombyx Batryticatus in Chinese Pharmacopoeia(2020). The results revealed that in the 102 batches of Bombyx Batryticatus, the qualification rate of silk gland was low; the content of total ash far exceeded the standard; the content of beauvericin varied greatly. The qualification rate of the silk gland of the 35 batches of Bombyx Batryticatus was only 47.49%, which could be increased to 73.00% if the number of silk gland was 2 to 4. The death rate of Bombyx Batryticatus at different infection ages was quite different, with uneven quality. Generally, the yield of Bombyx Batryticatus inoculated on the first day of the fifth instar was high with good quality. Therefore, in combination with the quality and actual production of Bombyx Batryticatus, the following suggestions were proposed for revision of Bombyx Batryticatus in Chinese Pharmacopoeia(2025): The number of silk gland should be revised as 2-4 bright brown or bright black silk glands, after which, the quality of Bombyx Batryticatus could be guaranteed, and the "quality identification based on character" could also be reflected scientifically; the content determination index that the content of beauvericin shall not be less than 0.017% should be added to better control the quality of Bombyx Batryticatus; the infection age should be revised as the first day of the fifth instar to narrow the age span, which could better fit the actual production and ensure the quality of Bombyx Batryticatus.
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Animais , Bombyx , Medicina Tradicional do Leste Asiático , Seda , LarvaRESUMO
OBJECTIVES@#This study aims to construct endogenous exosomes abundantly loaded with miR-1 and investigate the role of exosome-mediated microRNA-1 (miR-1) delivery on CAL-27 cell proliferation.@*METHODS@#Exosomes secreted by miR-1-overexpressing HEK293 cells (miR1-EXO) were purified via ultracentrifugation and subjected to transmission electron microscopy, nanoparticle analysis, Western blot analysis, and quantitative polymerase chain reaction (qPCR). CAL-27 cells were cocultured with exosomes secreted by HEK293 cells (CON-EXO) and miR1-EXO and equivalent phosphate buffer saline. The intracellular transport of exosomes was measured by using immunofluorescence, the expression of miR-1 and its target gene MET were investigated via qPCR, CAL-27 cell proliferation was measured through MTT assay, and cell cycle state was determined by applying flow cytometry.@*RESULTS@#Electron microscopy revealed that miR1-EXO and CON-EXO were spherical or cup-shaped with an average diameter of approximately 110 nm. The well-known exosome markers CD9, Tsg101, and Alix were enriched. The expression of miR-1 in miR1-EXO was higher than that in CON-EXO (285.80±14.33 vs 1.00±0.06, @*CONCLUSIONS@#Exosomes secreted from miR1-EXO cells could load abundant miR-1. Exosomal miR-1 delivered into CAL-27 cells by using miR1-EXO suppressed the expression of MET mRNA and inhibited cell proliferation.
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Humanos , Ciclo Celular , Proliferação de Células , Exossomos , Células HEK293 , MicroRNAsRESUMO
Objective@#To analyze the differentially expressed genes related to the chemosensitivity with the TPF regimen for hypopharyngeal squamous cell carcinoma and to measure potential functional targeting genes expressions.@*Methods@#Twenty-nine patients with primary hypopharyngeal cancer who underwent induction chemotherapy with TPF from January 2013 to December 2017 in Beijing Tongren Hospital were enrolled for microarray analysis, including 28 males and 1 female, aged from 43 to 73 years old. Among them, 16 patients were sensitive to chemotherapy while 13 patients were non-sensitive. Illumina Human HT-12 Bead Chip was applied to analyze the gene expressions and online bioinformatics analysis was used to analyze the differentially expressed genes. Reverse transcription and quantitative real-time PCR (RT-qPCR) was used to measure the mRNA expression of potential functional genes of TPF induction chemotherapy in 43 samples, 29 from original patients and 14 from additional patients. Graphpad prism 7.0 software was used for statistical analysis.@*Results@#A total of 1 381 significantly differentially expressed genes were screened out. By GO analysis, up-regulated genes included sequestering in extracellular matrix, chemokine receptor binding and potassium channel regulator activity; down-regulated genes included regulation of angiogenesis, calcium ion binding and natural killer cell activation involved in immune response. With KEGG database analysis, down-regulated pathways included ECM-receptor interaction and peroxisome and up-regulated pathways included Glutathione metabolism and PPAR signaling pathway. The expressions of CD44 and IL-6R were significantly different and appeared biologically significant. CD44 was significantly upregulated in insensitive tissues (0.54±0.06) compared with sensitive tissues (0.33±0.04)(P<0.01). IL-6R was significantly downregulated in insensitive tissues (0.44±0.03) compared with sensitive tissues. (0.68±0.03) (P<0.01).@*Conclusion@#CD44 and IL-6R may be potentially functional genes of TPF induction chemotherapy in hypopharyngeal squamous cell carcinoma.
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To analyze the differentially expressed genes related to the chemosensitivity with the TPF regimen for hypopharyngeal squamous cell carcinoma and to measure potential functional targeting genes expressions. Twenty-nine patients with primary hypopharyngeal cancer who underwent induction chemotherapy with TPF from January 2013 to December 2017 in Beijing Tongren Hospital were enrolled for microarray analysis, including 28 males and 1 female, aged from 43 to 73 years old. Among them, 16 patients were sensitive to chemotherapy while 13 patients were non-sensitive. Illumina Human HT-12 Bead Chip was applied to analyze the gene expressions and online bioinformatics analysis was used to analyze the differentially expressed genes. Reverse transcription and quantitative real-time PCR (RT-qPCR) was used to measure the mRNA expression of potential functional genes of TPF induction chemotherapy in 43 samples, 29 from original patients and 14 from additional patients. Graphpad prism 7.0 software was used for statistical analysis. A total of 1 381 significantly differentially expressed genes were screened out. By GO analysis, up-regulated genes included sequestering in extracellular matrix, chemokine receptor binding and potassium channel regulator activity; down-regulated genes included regulation of angiogenesis, calcium ion binding and natural killer cell activation involved in immune response. With KEGG database analysis, down-regulated pathways included ECM-receptor interaction and peroxisome and up-regulated pathways included Glutathione metabolism and PPAR signaling pathway. The expressions of CD44 and IL-6R were significantly different and appeared biologically significant. CD44 was significantly upregulated in insensitive tissues (0.54±0.06) compared with sensitive tissues (0.33±0.04)(0.01). IL-6R was significantly downregulated in insensitive tissues (0.44±0.03) compared with sensitive tissues. (0.68±0.03) (0.01). CD44 and IL-6R may be potentially functional genes of TPF induction chemotherapy in hypopharyngeal squamous cell carcinoma.
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OBJECTIVE To evaluate the survival and functional outcomes of advanced laryngeal squamous cell carcinoma after modified supracricoid partial laryngectomy(SCPL).METHODS We selected 65 laryngeal cancer patients treated with modified supracricoid partial laryngectomy from 2002 to 2015 in our hospital.Among them,62 cases were males,3 cases were females with an age ranged from 35 to 80 years(median age 58 years)There were 26 cases with T2 stage,32 cases with T3 stage and 7 cases with T4 stage.We selected 120 laryngeal cancer patients treated with vertical partial laryngectomy at the same period as control.Then we evaluated the functional outcomes of modified supracricoid partial laryngectomy group compared with vertical partial hemilaryngectomy group.RESULTS The 5-year cumulative survival rate and decannulation rate were 82.3％ and 98.3％ for supracricoid laryngectomy group respectively.Decannulation ratewas 86.1％ for vertical partial hemilaryngectomy group(P＜0.05).However,there was no significant difference between supracricoid partial laryngectomy group and vertical partial laryngectomy group in pronunciation evaluation and abnormal deglutition.CONCLUSION Modified supracricoid partial laryngectomy is a good choice for local advanced laryngeal squamous cell carcinoma.
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Objective@#To establish a laryngeal squamous cell carcinoma (LSCC) cell line through primary cell culture and observe its biological characteristics.@*Methods@#Tissue block culture method was used for primary cell culture. After LSCC cells passed 25 times in vitro, the morphology of cells was observed, keratin was stained histochemically, cell cycle was tested by PI-FACS, and the specie of cells was detected by PCR and short tandem repeat(STR) typing.@*Results@#This newly established LSCC cell line was named as TR-LCC-1, most of the cancer cells were polygonal shape, like the cobblestone, loss of contact inhibition and with overlapping growth. Cell size was large and cell pleomorphism was very obvious. Cytokeratin staining was positive. After 6 months of continuous culture in vitro, the TR-LCC-1 cells passed more than 30 times, and cell doubling time was 201.2h. Cell cycle assay indicated that G1 phase accounted for 51.71%, S phase was 44.56%, and G2 phase was 2.28%. Mycoplasma test showed no mycoplasma contamination. Cell species identification identified TR-LCC-1 was human-derived cells. STR detection showed P26 and P6 were same, and they were different from the STR typing of disclosed cells.@*Conclusion@#The establish ment of the new laryngeal squamous carcinoma cell line TR-LCC-1 can be helpful to the research for laryngeal squamous cell cancer.
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[ABSTRACT]OBJECTIVEThe purpose of this study was to analyze the screened miRNAs related to the chemosensitivity for the TPF regimen of hypopharyngeal squamous cell carcinoma by miRNA array, and provide a set of miRNAs that may be useful for the development of novel diagnostic markers and more effective therapeutic strategies from the screened miRNAs.METHODSA total number of 21 patients who underwent TPF induction chemotherapy for primary hypopharyngeal squamous cell carcinoma were recruited for miRNA array analysis. 12 patients are sensitive to chemotherapy, and 9 patients are not. Moreover, the selected putative regulated miRNAs were also validated by RT-PCR in another 24 patients (14 patients are sensitive to chemotherapy, and others are not).RESULTSThere were 24 miRNA significantly differencial to the sensitivity to chemotherapy, and 6 miRNAs were up-regulated in the TPF group while 18 miRNA were down-regulated (P<0.05). To identify typical miRNA, mirfocus 3.0 database selected four miRNAs hsa-miR-211-3p, hsa-miR-4253, hsa-miR-4443, and hsa-miR-193b-3p, which were significant down-regulated in TPF-sensitive group. QRT-PCR further validated that only three miRNA (hsa-miR-4253、hsa-miR-4443、hsa-miR-193b-3p) were under-expressed in TPF-sensitive group of another 24 tissue samples (P<0.05).CONCLUSIONMiRNA hsa-miR-193b-3p, hsa-miR-4253, hsa-miR-4443 were identified in TPF-sensitive tissues by microarrays, and further validated by RT-PCR. These down-regulated miRNAs may act as novel biomarkers to classify TPF sensitivity of hypopharyngeal squamous cell carcinoma patients and will contribute to the understanding of the molecular basis of the chemosensitivity in the disease.
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Objective To evaluate the efficacy of laryngeal mask airway (LMA) Ⅰ-gel for airway management in the patients requiring insertion of nasogastric tube before laparoscopic cholecystectomy.Methods Sixty patients,aged 26-64 yr,weighing 54-90 kg,of ASA physical status Ⅰ-Ⅲ (Mallampati Ⅰ-Ⅲ),scheduled for elective laparoscopic surgery,were randomly divided into Ⅰ,Ⅱ and Ⅲ groups (n =20 each) using a random number table.In group Ⅰ,the nasogastric tube was inserted through the drain tube of LMA I-gel.In group Ⅱ,the nasogastric tube was inserted through the nostril before surgery.In group Ⅲ,the nasogastric tube was inserted through the nostril before surgery,and another nasogastric tube was inserted through the drain tube of LMA I-gel after induction of anesthesia.The hemodynamic parameters,SpO2,PET CO2 and peak airway pressure were monitored during surgery.The fiberoptic laryngoscopy scores were assessed and the development of nasogastric tube displacement was recorded after successful LMA placement.The LMA placement time,success rate of LMA placement at the first attempt,depth of placement,airway sealing pressure,and occurrence of air leakage of LMA and nasogastric tube drainage were recoded.The bloodstains and gastroesophageal reflux were observed after removal of LMA Ⅰ-gel.The pH values were tested at the tip of LMA and on the dorsal and ventral sides of the body of LMA by using pH test papers.The development of adverse reactions in the oropharynx was recorded within 24 h after surgery.Results The hemodynamics was stable and SpO2,peak airway pressure and PETCO2 were all within the normal range during surgery,and Ppeak was lower than airway sealing pressure in the three groups.There were no significant differences between the three groups in LMA placement time,success rate of LMA placement at the first attempt,depth of placement,airway sealing pressure,incidence of air leakage of LMA,fiberoptic laryngoscopy scores,time for removal of LMA I-gel,incidences of adverse reactions in the oropharynx,bloodstains within the body of LMA and gastroesophageal reflux,and pH values at the tip of LMA and on the dorsal and ventral sides of the body of LMA.There was no nasogastric tube displacement in Ⅱ and Ⅲ groups.There were 7 patients developing gastric juice outflow from drainage tube of the LMA I-gel and 2 patients developing gastric juice outflow from the nostril gastrictubes in group Ⅱ.Conclusion For the patients requiring insertion of nasogastric tube before laparoscopic cholecystectomy,insertion of LMA I-gel is easy,and I-gel LMA can assure good airway sealing and adequate ventilation.
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<p><b>OBJECTIVE</b>To identify the genes differentially expressed and underlying molecular mechanism in laryngeal squamous cell carcinoma by using cDNA microarrays.</p><p><b>METHODS</b>Using Illumina Human HT-12 BeadChip, gene expressions were detected in ten pairs of laryngeal cancer tissues and adjacent normal tissues. Total RNA was extracted and reverse-transcribed into cDNA. Labeled cDNA were hybridized with cDNA microarray, data were read and images were scanned. All the samples had passed quality control testing.</p><p><b>RESULTS</b>Through Illumina Genomstudio 1.9.0 Data processing software (P < 0.05 or diffscore>13, diffscore<-13) and multiple displacement t test(FDR<0.05), 426 genes showed statistically significant differences in expressions between laryngeal tumor tissues and corresponding adjacent normal tissues, with 222 up-regulated genes and 204 down-regulated genes in laryngeal cancer tissues. These up- or down-regulated genes were indicated to involve in cellular processes relevant to the cancer phenotype, such as proliferation, cell cycle, chromosome segregation, mitosis and meiosis. These differentially expressed genes also took part in cancer related signaling pathways as well, for instance, metabolic pathways, cell cycle, DNA replication, glutathione metabolism, mucin type O-Glycan biosynthesis, drug metabolism-cytochrome P450 and so on.</p><p><b>CONCLUSION</b>The set of genes identified here and their functional annotations contribute to a better understanding of the pathogenesis of laryngeal squamous cell carcinoma from the view of multiple gene interactions and provide candidate markers for improving diagnosis, prognosis and treatment of this cancer.</p>
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Idoso , Idoso de 80 Anos ou mais , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma de Células Escamosas , Genética , Expressão Gênica , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Estudo de Associação Genômica Ampla , Neoplasias de Cabeça e Pescoço , Genética , Neoplasias Laríngeas , Genética , Análise de Sequência com Séries de OligonucleotídeosRESUMO
<p><b>BACKGROUND</b>Gene therapy and epigenetic therapy have gained more attention in cancer treatment. However, the effect of a combined treatment of gene therapy and epigenetic therapy on head and neck squamous cell carcinoma have not been studied yet. To study the mechanism and clinical application, human laryngeal carcinoma cell (Hep-2) tumor-bearing mice were used.</p><p><b>METHODS</b>A xenograft tumor model was established by the subcutaneous inoculation of Hep-2 cells in the right armpit of BALB/c nu/nu mice. The mice with well-formed tumor were randomly divided into six groups. Multisite injections of rAd-p53 and/or 5-aza-dC were used to treat tumor. Tumor growth was monitored by measuring tumor volume and growth rate. p53 and E-cadherin protein levels in tumor tissues were detected by immunohistochemical staining. The mRNA levels were monitored with FQ-PCR.</p><p><b>RESULTS</b>Gene therapy was much more effective than single epigenetic therapy and combined therapy. The gene therapy group has the lowest tumor growth rate and the highest expression levels of p53 and E-cadherin.</p><p><b>CONCLUSIONS</b>The combined treatment of gene and epigenetic therapy is not suggested for treating head and neck carcinoma, because gene therapy shows an antagonistic effect to epigenetic therapy. However, the mechanisms of action are still unclear.</p>
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Animais , Humanos , Masculino , Camundongos , Azacitidina , Usos Terapêuticos , Caderinas , Metilases de Modificação do DNA , Epigênese Genética , Genes p53 , Terapia Genética , Neoplasias Laríngeas , Genética , Patologia , Terapêutica , Camundongos Endogâmicos BALB C , Proteína Supressora de Tumor p53 , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
<p><b>OBJECTIVE</b>To observe the effects of gene therapy and epigenetic therapy on the tumor growth of laryngeal carcinoma and the underlying mechanisms.</p><p><b>METHODS</b>The animal model of human laryngeal carcinoma was established by the subcutaneous inoculation of Hep-2 cells at the right armpit of BALB/c nu/nu mice. The tumor-bearing mice were randomized into 4 groups, p53 therapy group(rAd-p53), epigenetic therapy group(5-aza-dC), combination therapy group (rAd-p53+5-aza-dC) and control group. The gene and protein expressions of molecular markers p53 and E-cadherin were detected by FQ-PCR and immunohistochemistry.</p><p><b>RESULTS</b>By the day 20 of the treatments, the mean tumor volumes were(106.09 ± 24.40)mm(3) in p53 therapy group, (166.55 ± 40.11) mm(3) in epigenetic therapy group, (126.11 ± 22.49) mm(3) in combination therapy group,and (252.83 ± 54.09) mm(3) in control group. Both gene therapy (F = 37.30, P < 0.05) and epigenetic therapy (F = 4.79, P < 0.05) inhibited the growth of xenografted tumors, with an interaction effect (F = 22.01, P < 0.05) between the two groups. The integral optical density value of p53 protein expression of p53 therapy group (628.07 ± 95.16) was significantly higher than that of combination therapy group (494.76 ± 100.22), (t = 8.72, P < 0.05). The integral optical density values of E-cadherin protein expression were 558.89 ± 97.58 in p53 therapy group, 380.41 ± 90.60 in epigenetic therapy group, 494.76 ± 102.88 in combination therapy group,and 162.60 ± 40.38 in control group respectively, indicating the enhancements of E-cadherin protein expression by gene therapy (F = 45.24, P < 0.05) or epigenetic therapy(F = 5.73, P < 0.05)and the existence of interaction effect (F = 21.82, P < 0.05) between gene therapy and epigenetic therapy. The expression levels of p53 gene were 4.43 ± 0.12 in p53 therapy group, 1.06 ± 0.11 in epigenetic therapy group, 3.51 ± 0.10 in combination therapy group,and 1.09 ± 0.11 in control group, respectively, showing an interaction effect between gene therapy and epigenetic therapy (F = 298.11, P < 0.05). The expression levels of E-cadherin gene were 4.50 ± 0.34 in p53 therapy group, 2.02 ± 0.16 in epigenetic therapy group, 2.99 ± 0.12 in combination therapy group, and 1.00 ± 0.11 in control group, respectively. The expression of E-cadherin gene was enhanced by gene therapy (F = 329.12, P < 0.05)or epigenetic therapy(F = 88.57, P < 0.05), with an interaction effect between the two therapies (F = 122.17, P < 0.05).</p><p><b>CONCLUSIONS</b>Xenografted tumors of human laryngeal carcinoma cells are inhibited by gene therapy, the epigenetic therapy and the combination therapy. The gene therapy was significantly better than the epigenetic therapy or the combination therapy. There might be antagonistic effect between p53 and 5-aza-dC.</p>
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Animais , Humanos , Masculino , Camundongos , Caderinas , Metabolismo , Carcinoma , Terapêutica , Linhagem Celular Tumoral , Terapia Combinada , Epigenômica , Terapia Genética , Neoplasias Laríngeas , Terapêutica , Camundongos Endogâmicos BALB C , Camundongos Nus , Transplante de Neoplasias , Proteína Supressora de Tumor p53 , MetabolismoRESUMO
Along with the improvement of the diagnosis techniques and the popularization of the health examination,the incidence of the thyroid nodule is growing rapidly.The management methods of the nodules depend on the correct diagnosis of the thyroid nodules,which could save the limited resources of the healthcare system.The diagnostic methods of the thyroid nodules developed in recent years are reviewed in this paper.