miR-100-5p对甲状腺癌细胞增殖与凋亡调控作用的实验研究

目的通过实验探讨微小核糖核酸(microRNA,miR)-100-5p在甲状腺癌细胞中的表达情况及其对细胞增殖与凋亡的调控作用。方法使用荧光定量PCR检测miR-100-5p在甲状腺癌细胞系(TPC-1,KTC-1)与甲状腺正常细胞系(Nthy-ori3-1)中的相对表达情况。TPC-1细胞分别转染miR-100-5p模拟物(miR-100-5p mimic)、抑制物(miR-100-5p inhibitor)及相应阴性对照(miR-mimic NC,miR-inhibitor NC)后,用CCK-8检测TPC-1细胞增殖情况,流式细胞仪检测TPC-1细胞凋亡情况。通过miRTarBase和TargetScan7.2数据库对miR-100-5p的靶基因进行预测和功能富集分析,用蛋白印迹实验与双荧光素酶报告基因实验验证miR-100-5p对成纤维细胞生长因子受体3(fibroblast growth factor receptor 3,FGFR3)的靶向调控作用。结果与Nthy-ori3-1细胞相比,miR-100-5p在TPC-1细胞中表达水平(1.87±0.03 vs 1.00±0.03)与KTC-1细胞中表达水平(6.33±0.47 vs 1.00±0.03)均上调,差异具有统计学意义(t=-34.220,-19.588,均P<0.05)。转染miR-100-5p mimic组在24,48,72h细胞450nm吸光度(A_(450nm))均高于miR-mimic NC组,差异具有统计学意义(t=-7.516,-17.828,-8.445,均P<0.05);转染miR-100-5p inhibitor组在24,48,72h A_(450nm)均低于miR-inhibitor NC组,差异具有统计学意义(t=6.720,6.782,6.073,均P<0.05)。与miR-mimic NC组相比,转染miR-100-5p mimic后凋亡率(7.43%±0.49%vs 10.55%±0.80%)下降(t=5.767,P=0.004),与miR-inhibitor NC组相比,转染miR-100-5p inhibitor后凋亡率(3.19%±0.22%vs 2.64%±0.15%)上升(t=-3.606,P=0.023),差异均有统计学意义。蛋白印迹实验显示,与miR-mimic NC组相比,FGFR3在miR-100-5p mimic组蛋白表达水平(0.78±0.12 vs 1.00±0.00)下调(t=3.071,P=0.037),与miR-inhibitor NC组相比,FGFR3在miR-100-5p inhibitor组蛋白表达水平(1.17±0.07 vs 1.00±0.00)上升(t=-4.509,P=0.046),差异均有统计学意义。与miR-mimic NC相比,miR-100-5p mimic没有降低FGFR33’UTR野生型组荧光素酶活性(1.01±0.17 vs 1.00±0.00)与突变型组荧光素酶活性(0.99±0.11 vs 1.00±0.00),差异无统计学意义(t=-0.057,0.181,P=0.96,0.873)。结论miR-100-5p在甲状腺癌细胞中表达上调,可促进甲状腺癌细胞增殖、抑制细胞凋亡,其可能成为甲状腺癌诊疗中新的生物标志物与调控靶点。

S100A4 silencing blocks invasive ability of esophageal squamous cell carcinoma cells

AIM： To investigate a potential role of S100A4 in esoph- agus squamous cell carcinoma metastasis （ESCCs）.METHODS： Expression of $100A4 and E-cadherin were analyzed in frozen sections from ESCCs （metastasis, n = 28; non-metastasis, n = 20） by reverse transcrip- tion-polymerase chain reaction, quantitative polymerase chain reaction and immunohistochemistry. To explore the influence of $100A4 on esophageal cancer invasion and metastasis, $100A4 was overexpressed or silenced by $100A4 siRNA in TE-13 or Eca-109 cells/n vitro and /n vivo.

Single-cell transcriptome profiling of sepsis identifies HLA-DR^(low)S100A^(high)monocytes with immunosuppressive function

Background Sustained yet intractable immunosuppression is commonly observed in septic patients,resulting in aggravated clinical outcomes.However,due to the substantial heterogeneity within septic patients,precise indicators in deciphering clinical trajectories and immunological alterations for septic patients remain largely lacking.Methods We adopted cross-species,single-cell RNA sequencing(scRNA-seq)analysis based on two published datasets containing circulating immune cell profile of septic patients as well as immune cell atlas of murine model of sepsis.Flow cytometry,laser scanning confocal microscopy(LSCM)imaging and Western blotting were applied to identify the presence of S100A9^(+)monocytes at protein level.To interrogate the immunosuppressive function of this subset,splenic monocytes isolated from septic wild-type or S100a9^(–/–)mice were co-cultured with naive CD4^(+)T cells,followed by proliferative assay.Pharmacological inhibition of S100A9 was implemented using Paquinimod via oral gavage.Results scRNA-seq analysis of human sepsis revealed substantial heterogeneity in monocyte compartments following the onset of sepsis,for which distinct monocyte subsets were enriched in disparate subclusters of septic patients.We identified a unique monocyte subset characterized by high expression of S100A family genes and low expression of human leukocyte antigen DR(HLA-DR),which were prominently enriched in septic patients and might exert immunosuppressive function.By combining single-cell transcriptomics of murine model of sepsis with in vivo experiments,we uncovered a similar subtype of monocyte significantly associated with late sepsis and immunocompromised status of septic mice,corresponding to HLA-DR^(low)S100A^(high)monocytes in human sepsis.Moreover,we found that S100A9^(+)monocytes exhibited profound immunosuppressive function on CD4^(+)T cell immune response and blockade of S100A9 using Paquinimod could partially reverse sepsis-induced immunosuppression.Conclusions This study identifies HLA-DR^(low)S100A^(high)monocytes correlated with immunosuppressive state upon septic challenge,inhibition of which can markedly mitigate sepsis-induced immune depression,thereby providing a novel therapeutic strategy for the management of sepsis.

Preparation of acellular nerve grafts with triton X-100

BACKGROUND: The source of nerve allograft enriches. We may choose expediently nerve allograft to repair injured nerve and the structure of choice nerve homology or similar with the injured nerve, but the immunological rejection limits the clinical application of nerve allograft. The ideal substitute of autograft never is researching. OBJECTIVE: In this experiment, Triton X-100 was used to extract the Schwann cells and myelin sheaths of allograft nerve and obtain the inartificial and eliminated antigenicity nerve-transplanter (nerve grafts). DESIGN: Controlled experiment. SETTING: Department of Hand Surgery, the Third Affiliated Hospital of Hebei Medical University; Second Department of Orthopedics, Fourth Center Hospital of Tianjin. MATERIALS: Thirty health New Zealand big ear white rabbit, of either sex (gender), weighing 2000-3000 g, were provided by the Center of Experimental Animal of Hebei Medical University. TritonX-100 was offered by SIGMA Company. METHODS: The experiment was carried out at the Central Laboratory of the Third Affiliated Hospital of Hebei Medical University from December 2003 to December 2004. Sixty pieces of sciatic nerves, 10-mm-long nerve segment, which were taken from 30 rabbits, were incised. They were randomly divided into chemical extraction group (n =50) and control group (n =10). In the chemical extraction groups, the nerves were put into 3% Triton X-100 solution. They were treated with Triton X-100 for 12 hours, 24 hours, 48 hours, 96 hours and 1 week, respectively. They were examined in every period. The control groups did not treated with anything. ① Respectively two segments of nerve by 2 mm length were taken from each nerve in the every periods. ② The laminin immunohistochemical stained sections were performed with image acquisition and analyzed with multicolor pathological image analysis system. Measured the laminin antibody reaction part of each section and computed laminin average gray degrees of the unit area. All dates were analyzed by SPSS 10.0 software. MAIN OUTCOME MEASURES: ① General observation and histological observation in two groups; ② Compared with laminin average gray degrees of the unit area in each section. RESULTS: ① General observation: In the control groups, fresh nerve was polish, rigidity and elasticity. After the nerves were chemical extracted, the floccules was seen at two ends and around of the nerves. The nerves being extraction presented ivory and lackluster. Its diameter and length compared reduced, tenderness and tenacity with the fresh nerve. Observed by light microscope, Schwann cells, myelin sheaths and basement membrane distribute uniformly in control groups. After the nerves were extracted, Schwann cells and myelin sheaths disappeared. Basement membrane presented barrier array in longitudinal sections. Between the membranes was the basement membrane tube. Observed with scanning electron microscope, the basement membrane tubes composed by collagen fibers were remained and collagen fibers maintained their former position, form and structure. Further, the structure of membrane was seen in the tubes. It was Schwann cells basement membrane. ② In chemical extraction groups, laminin average gray degrees of the unit area were 140.1±3.41 (12 hours), 142.1±3.14 (24 hours), 142.1±3.14 (48 hours), 140.4±4.03 (96 hours), 141.7±2.62 (1 week). In the control groups, laminin average gray degree of the unit area was 142.7±7.24. There were not significant differences among the groups (P > 0.05). CONCLUSION: The method of chemical extraction by using of Triton X-100 may be an ideal measure for preparing tissue-engineered nerve-transplanter and reserved the live of laminin in the basement membrane.

S100 protein expression during induced Schwann cell-like cell differentiation of rat bone marrow mesenchymal cells in vitro

BACKGROUND： S100 protein can promote axonal growth. Therefore, transplantation of induced bone marrow-derived mesenchymal stem cells （MSCs） that can secrete S100 may provide a beneficial microenvironment for neural regeneration. OBJECTIVE： To explore the changes in S100 expression during rat MSCs differentiation into Schwann ceils in vitro. DESIGN, TIME AND SETTING： This cytology experiment was performed at the Jiangsu Key Laboratory of Neuroregeneration, Nantong University in China, from January 2006 to May 2007. MATERIALS： The rabbit anti-S100 polyclonal antibody was purchased from Dako, Denmark; the mouse anti-rat S100 monoclonal antibody was purchased from Sigma, USA. METHODS： MSCs were cultured from adult Sprague-Dawley rat femur and tibia. Cell proliferation was determined by the MTT method and CD markers, and cell cycle was measured by flow cytometry. MSCs were induced to differentiate into SC cells. SC cells were stained for S100 protein, glial fibrillary acidic protein, and low-affinity nerve growth factor receptor. S100 protein and mRNA levels were evaluated by flow cytometry, Western blot, and reverse transcription-polymerase chain reaction. MAIN OUTCOME MEASURES： S100 protein and mRNA expression. RESULTS： MSCs exhibited high amplification potential over eight passages. Prior to induction, the majority of MSCs were at the G0/G1 phase of the cell cycle. After induction, MSCs displayed morphology changes similar to Schwann cells. Moreover, induction increased S100 mRNA levels. Immunofluorescence showed that MSCs expressed S100 protein, glial fibrillary acidic protein, and low-affinity nerve growth factor receptor at 7 days of induction. Induction also increased S100 protein levels compared with untreated MSCs. CONCLUSION： MSCs are capable of differentiating into Schwann cells-like cells under conditional induction in vitro, with increasing S100 mRNA and protein expression.

S100 PROTEIN-POSITIVE DENDRITIC CELLS AND THE SIGNIFICANCE OF THEIR DENSITY IN GASTRIC PRECANCEROUS LESIONS

Quantitative analysis of dendritic cells (DC’s) was carried out in tissue specimens of normalgastric mucosa (n=15),gastric ulcer (n=19),chronic atrophic gastritis (n=28),and gastriccarcinoma (n=65) by ABC immunostaining with S100 protein antibody.Significant increasein DC number were observed in chronic atrophic gastritis with type Ⅲ intestinal metaplasiaand/or grade Ⅱ,Ⅲ dysplasia.The result suggests that DC’s are potentially capable opresenting neoantigens associated with malignant transformation at the precancerous stagewhen malignant morphological changes have not yet taken place.Combined with routinediagnostic methods,the serial monitoring of DC density in gastric mucosa may be usefulin the follow-up of premalignant lesions in the stomach and the diagnosis of early gastriccarcinoma.

Construction of eukaryotic expression vector of human S100A13 gene and its effect on proliferation of human thyroid cancer cell line TT

Objective To investigate the effect of exogenous S100A13 gene overexpression on the proliferation of human thyroid cancer cell line TT.Methods The recombinant ORF of S100A13 tagged with six histidines at the 5' end was subcloned into the pcDNA3.2/V5/GW/D-TOPO vector and sequenced.The eukaryotic expression plasmid pcDNA3.2/V5 /GW/D-S100A13 and empty vector pcDNA3.2/V5/GW/D were transfected into TT cells.The positive clones were selected by G418.The expressions of S100A13 mRNA and protein were detected by real time reverse transcription-polymerase chain reaction(RT-PCR) and Western blot.The effect of S100A13 on cell proliferation and cell cycle was evaluated by cell growth curve,MTT colorimetric assay and flow cytometry.Results S100A13 gene tagged with six histidines at the 5 ' end was confirmed to be inserted into the pcDNA3.2/V5/GW/D vector correctly.TT-S100A13-V5 cells,which over-expressed S100A13,were constructed successfully.TT-S100A13-V5 cells grew much faster than TT-V5 and TT cells(P <0.001).The proportions of both S and G2/M phase cells were significantly higher in TT-S100A13-V5 cells than those in TT-V5 and TT cells(P <0.001).Conclusion The eukaryotic expression vector containing human S100A13 gene has been successfully constructed,which highly expresses S100A13 in TT cells.Exogenous S100A13 gene overexpression accelerates TT cell proliferation and drives the cell cycle progression of TT cells from G0/G1 phase to S and G2/M phases.

S100A6 gene have a positive influence on the growth and proliferation of gastric cancer cell MKN45

Objective： S100A6 （a.k.a., calcyclin） is over-expressed in several human tumors, including gastric carcinoma, human melanoma, pancreatic carcinoma, squamous cell carcinoma, malignant fibrous histiocytoma （MFH）, and carcinomas of the thyroid, breast, and colon. However, little is known about the role S100A6 plays in gastric adenocarcinoma. In the present study, we intended to investigate the influence of S100A6 on the growth, proliferation, apoptosis, invasion and cell cycle of the gastric cancer cell MKN45. Methods： As an important member of S100 family, S100A6 cDNAwas subcloned into a constitutive vector pcDNA3.1 followed by transfection in gastric cancer cell line MKN45 by using liposome. Then stable transfectants were selected and appraised. The apoptosis and cell cycles of these clones were analyzed by using flow cytometric assay. The growth and proliferation were analyzed by cell growth curves and colony-forming assay respectively. The S100A6 stable expression clones （MKN-S100A6） were detected and compared with their control groups respectively. Results： MKN- S100A6 grew faster than MKN45 and MKN-PC （MKN45 transfected with pcDNA3.1 vector）. The cell counts of MKN-SI00A6 in the fifth, sixth and seventh days were significantly more than those of control groups （P 〈 0.05）. Cell cycle analysis showed that proportions of MKN-S100A6 in G0-G1 and G2-M were different significantly with those of its control groups respectively （P 〈 0.05）. The apoptosis rate of MKN-S100A6 was significantly lower than those of control groups （P 〈 0.05）. Results of colony-forming assay showed that the colon formation rate of MKN-S100A6 was higher than those of control groups （P 〈 0.05）. Conclusion： S100A6 can promote the growth and proliferation of gastric cancer cells. It can help tumor cell maintain malignant phenotype. In gastric cancer, S100A6 could be thought as a tumor-enhancing gene in some distance, but its role could be complicated.

S100A7 Co-localization and Up-regulation of Filaggrin in Human Sinonasal Epithelial Cells

Objective:Filaggrin(FLG)is a protein expressed in the epidermis and involved in the maintenance of the epidermal barrier.However,the expression and localization of FLG in the upper airway remain controversial.The present study aimed to determine the significance of FLG and the effect of S100A7 on FLG expression in the upper respiratory mucosa.Methods:Human nasal epithelial cells(HNECs)were cultured and examined for FLG expression and S100A7 effects by real-time polymerase chain reaction and Western blotting.The localization and distribution of FLG were assessed using sinonasal mucosa.Results:A significant expression of FLG was detected at the mRNA and protein levels in HNECs.A moderate FLG immunoreactivity was observed in the epithelial cells,but no staining was seen in epithelial goblet cells.S100A7 increased the FLG mRNA level in HNECs in a dose-dependent manner and also up-regulated the FLG protein in a dose-dependent manner.Conclusion:This study significantly contributes to a better understanding of the role of FLG in the pathogenesis of airway inflammation from the viewpoint of the epithelial barrier function.FLG-related events in response to S100A7 protein may represent novel therapeutic targets for the treatment of upper airway inflammation.

ASSAY OF S100+ LANGERHANS' CELLS IN CERVICAL CARCINOMA

Langerhans cells (LCs) in the normal cervix (control, 19 cases), cervical carcinoma in situ( CIS, 19 cases) and invasive cervical carcinoma (30 cases), interdigitatlng cells (IDCs) and follicular dendritic cells (FDCs) in obturator lymph node draining Invasive cervical carcinoma were quantitatively assessed by ABC immunohistochemical staining using anti-S100 protein antibody. The results indicated that S100+ LCs in sltu, S100+ IDCs and S100+ FDCs In obturator lymph node showed dendritic features with a specific distribution. Number of LCs in situ in Invasive carcinoma increased significantly when compared with CIS and control groups. There was no change in LCs number between grade Ⅱ and Ⅲ of squemous cell carcinoma. Number of IDCs was significantly less in stage Ⅱ than in stage Ⅰ . FDCs number In different Invasive depthes and clinical stages showed no obvious change. The results suggest that progression of cervical carcinoma is closely related to decrease of LCs hi situ and IDCs in regional lymph node. Predominant Immune response of the host to cervical carcinoma should be cellular immunity.

IMMUNOHISTOCHEMICAL STUDY ON S100 PROTEIN-POSITIVE DENDRITIC CELLS IN PATIENTS WITH GASTRIC CARCINOMA

The density of dendritic cells (DC) and macro-phages (Mφ) in tissue specimens of gastric carcinoma (GC n=65) was investigated by ABC im-munohistochemical method using anti-S100 protein and anti-lysozyme antibodies, and was compared with that in gastric ulcer (GU n=19), chronic atrophic gastritis (CAG n=28) and normal gastric mucosa (NGM n=15). The mean density if DC (cells/mm2) in GC (15.0 was significantly higher than that in NGM (3.8) and GU (8.3), but was remarkably lower when compared to that in CAG (29.5) (P<0.01). Statistically significant difference in the population density of DC was observed between well- and poorly-differentiated GC (P<0.01). With their unique dendritic processes, DC were mainly concentrated within dense lymphoid infiltrates or in the T-area of reactive lymphoid follicles and were interspersed among the tumor cells. In contrast, Mφ were present around the necrotic foci and were rarely seen within the non-necrotic neoplastic tissues. These data suggest that DC, which differ in morphology, distribution, number and function form Mφ may be more directly involved in the host immune reaction against tumor by acting as antigen presenting cells.

Amyloid precursor-like protein 2 C-terminal fragments upregulate S100A9 gene and protein expression in BV2 cells

The murine microglial cell line BV2 has neuroprotective effects, but is toxic to neurons by secret-ing inlfammatory cytokines, and is an important target in the treatment of nerve inlfammation and neurodegenerative diseases. In the present study, we observed the effects of transfecting three amyloid precursor-like protein 2 （APLP2） C-terminal fragments （CTFs; C57, C50 and C31） in the pEGFP-N1 vector on S100A9 expression in BV2 cells. Reverse transcription-PCR, western blot assay and immunocytochemistry revealed that S100A9 protein and mRNA expression was greater in BV2 cells after CTF transfection than after mock transfection with an empty vector. Furthermore, transfection of full-length APLP2-751 resulted in low levels of S100A9 protein ex-pression. Our results show that APLP2-CTFs upregulate S100A9 protein and mRNA expression in BV2 cells, and identify a novel pathway involved in neuronal injury and apoptosis, and repair and protection in Alzheimer’s disease.

Correlation of S100A13 and FOXA1 expression with cell cycle and cell invasion in fine needle aspiration thyroid carcinoma tissue

Objective: To study the correlation of S100A13 and FOXA1 expression with cell cycle and cell invasion in fine needle aspiration thyroid carcinoma tissue. Methods: Patients who received ultrasound-guided thyroid nodule fine needle aspiration in Haiyang People's Hospital between April 2015 and February 2017 were selected, and the tissues were divided into malignant thyroid tissue and benign thyroid nodules according to the pathological results after biopsy. The expression of S100A13, FOXA1, cell cycle molecules and cell invasion molecules were measured. Results: S100A13, FOXA1, CDK2, CyclinD1, MCM2, MCM7, SKP2, CLOCK, STAT3, STAT5, N-cadherin, MT1-MMP and ADAM17 mRNA expression in thyroid carcinoma tissue were significantly higher than those in benign thyroid nodule;CDK2, CyclinD1, MCM2, MCM7, SKP2 and CLOCK mRNA expression in thyroid carcinoma tissue with high FOXA1 expression were significantly higher than those in thyroid carcinoma tissue with low FOXA1 expression;STAT3, STAT5, N-cadherin, MT1-MMP and ADAM17 mRNA expression in thyroid carcinoma tissue with high S100A13 expression were significantly higher than those in thyroid cancer tissue with low S100A13 expression. Conclusions: High expression of S100A13 and FOXA1 in thyroid carcinoma can promote cell invasion and cell cycle progression.

食管鳞状细胞癌病人血清miR-21-5p和miR-100-5p水平与临床特征和预后的相关分析

目的 分析食管鳞状细胞癌(ESCC)病人血清miR-21-5p和miR-100-5p的表达对ESCC鉴别诊断、临床特征和预后的影响。方法 2018年1月~2022年1月于我院放疗科接受放射治疗的158例晚期ESCC病人为实验组,150例健康体检志愿者为对照组。实时定量PCR(RT-qPCR)检测miR-21-5p和miR-100-5p表达水平。利用t检验比较miR-21-5p和miR-100-5p在实验组和健康对照组间的表达差异,采用ROC曲线分析其诊断价值。利用Kaplan-Meier法Log-Rank检验进行预后生存分析。多因素Cox回归分析ESCC病人预后的影响因素。结果 与对照组比较,ESCC病人血清miR-21-5p和miR-100-5p表达水平显著升高(P<0.001)。ROC曲线分析提示,二者联合诊断ESCC的AUC为0.918,灵敏度为93.2%,特异度为87.1%。ESCC病人血清miR-21-5p和miR-100-5p表达升高与更晚的TNM分期和更差的无进展生存期和总体生存期显著相关(P<0.05)。多因素Cox分析显示,TNM分期为Ⅲ期、血清中miR-21-5p高表达和miR-100-5p低表达是ESCC病人无进展生存期和总体生存期的独立危险因素(P均<0.05)。结论 血清miR-21-5p和miR-100-5p表达水平在ESCC病人中显著升高,且提示更差的预后。血清miR-21-5p和miR-100-5p,有望成为ESCC诊断和预后预测的生物标志物。

Regulation of TMEM100 expression by epigenetic modification,effects on proliferation and invasion of esophageal squamous carcinoma

BACKGROUND Esophageal squamous cell carcinoma(ESCC)is a prevalent malignancy with a high morbidity and mortality rate.TMEM100 has been shown to be suppressor gene in a variety of tumors,but there are no reports on the role of TMEM100 in esophageal cancer(EC).AIM To investigate epigenetic regulation of TMEM100 expression in ESCC and the effect of TMEM100 on ESCC proliferation and invasion.METHODS Firstly,we found the expression of TMEM100 in EC through The Cancer Genome Atlas database.The correlation between TMEM100 gene expression and the survival of patients with EC was further confirmed through Kaplan-Meier analysis.We then added the demethylating agent 5-AZA to ESCC cell lines to explore the regulation of TMEM100 expression by epigenetic modification.To observe the effect of TMEM100 expression on tumor proliferation and invasion by overexpressing TMEM100.Finally,we performed gene set enrichment analysis using the Kyoto Encyclopaedia of Genes and Genomes Orthology-Based Annotation System database to look for pathways that might be affected by TMEM100 and verified the effect of TMEM100 expression on the mitogen-activated protein kinases(MAPK)pathway.RESULTS In the present study,by bioinformatic analysis we found that TMEM100 was lowly expressed in EC patients compared to normal subjects.Kaplan-meier survival analysis showed that low expression of TMEM100 was associated with poor prognosis in patients with EC.Then,we found that the demethylating agent 5-AZA resulted in increased expression of TMEM100 in ESCC cells[quantitative real-time PCR(qRT-PCR)and western blotting].Subsequently,we confirmed that overexpression of TMEM100 leads to its increased expression in ESCC cells(qRT-PCR and western blotting).Overexpression of TMEM100 also inhibited proliferation,invasion and migration of ESCC cells(cell counting kit-8 and clone formation assays).Next,by enrichment analysis,we found that the gene set was significantly enriched in the MAPK signaling pathway.The involvement of TMEM100 in the regulation of MAPK signaling pathway in ESCC cell was subsequently verified by western blotting.CONCLUSION TMEM100 is a suppressor gene in ESCC,and its low expression may lead to aberrant activation of the MAPK pathway.Promoter methylation may play a key role in regulating TMEM100 expression.

金银花多酚粗提物调控miR-100-5p参与非小细胞肺癌细胞NCI-H1299侵袭迁移

目的:探讨金银花多酚粗对非小细胞肺癌NCI-H1299细胞恶性生物学行为的影响及分子机制。方法:将非小细胞肺癌NCI-H1299细胞分为NC组、紫杉醇组、金银花多酚粗提物低、中、高剂量组、miR-100-5p组、miR-NC组、anti-miR-100-5p+高剂量组、anti-miR-NC+高剂量组;四甲基偶氮唑盐比色法(MTT)检测细胞活性;Transwell检测细胞迁移和侵袭;蛋白质印迹法检测E-Cadherin、N-Cadherin、Vimentin和Wnt/β-catenin通路相关蛋白表达;实时荧光定量PCR(RT-qPCR)检测miR-100-5p表达水平。结果:不同浓度金银花多酚粗提物处理后,NCI-H1299细胞活性降低,细胞迁移和侵袭数量减少,E-Cadherin、miR-100-5p表达水平升高,N-Cadherin、Vimentin表达水平降低(P<0.05)。过表达miR-100-5p后,NCI-H1299细胞活性升高,细胞迁移[(226±20.03)个]和侵袭数量[(171±15.03)个]增加,E-Cadherin表达水平(0.34±0.03)降低,N-Cadherin(0.88±0.08)、Vimentin表达水平(0.95±0.09)升高(P<0.05)。下调miR-100-5p可逆转金银花多酚粗提物对NCI-H1299细胞增殖、迁移侵袭及Wnt/β-catenin通路的影响。结论:金银花多酚粗提物可能通过上调miR-100-5p表达,抑制Wnt/β-catenin通路活化,进而抑制非小细胞肺癌NCI-H1299细胞增殖、迁移和侵袭。

UF-100型尿沉渣检测仪测定红细胞影响因素的研究

目的 探讨草酸钙、非晶形盐、细菌、酵母菌在UF 10 0型尿沉渣全自动检测仪分析中对红细胞检测的干扰。方法 采用UF 10 0型尿沉渣全自动检测仪分析法、干化学分析法、镜检法共分析了 2 6 34份住院患者尿液标本 ,并将三者结果进行了比较。结果 单纯或混合性草酸钙、非晶形盐尿液标本 ,UF 10 0检测红细胞结果假阳性率为 95 % ;菌尿UF 10 0检测红细胞结果的假阳性率为 2 8% ;含酵母菌尿液UF 10 0检测红细胞结果假阳性率为 4 0 % ;正常对照组UF 10 0检测红细胞结果假阳性率为 5 %。与对照组相比 ,三者差异均有显著性 (P <0 .0 1)。结论 在UF 10 0型尿沉渣全自动检测仪分析中 ,草酸钙、非晶形盐、菌尿、酵母菌对红细胞检测的结果干扰很大 ,这对尿液检测质量控制具有重要的意义。将这 3种方法联合应用 ,可以提高检测结果的准确度 ,具有较高的临床应用价值。

苦参碱对K562细胞Cgi-100基因表达和细胞增殖的影响

本研究了解功能未知的cgi-100基因在苦参碱作用人白血病K562细胞中的表达情况,并探讨其对K562细胞生长的影响。应用RT-PCR检测苦参碱作用前后K562细胞中cgi-100基因的表达情况;构建pIRES2-EGFP/cgi-100真核表达质粒,用脂质体法转染至K562细胞中;RT-PCR检测在重组细胞中目的基因cgi-100的表达状况,并采用台盼蓝染色法观察细胞生长趋势,电镜观察细胞形态学改变,流式细胞术检测细胞周期的变化,观察cgi-100基因过表达对K562细胞增殖的影响。结果表明,在苦参碱作用K562细胞后cgi-100基因表达下调;重组K562细胞中cgi-100基因过表达,且常染色质增加,异染色质减少,细胞增殖旺盛,S期细胞增多,增殖速度高于对照组。结论:不同浓度不同作用时间下苦参碱能使K562细胞中cgi-100基因表达下降;cgi-100基因过表达能使K562细胞增殖加速、细胞幼稚程度增加。

S-100蛋白、CD1a、CD83及Ki-67在口腔朗格汉斯细胞组织细胞增生症中的表达及意义

目的对口腔朗格汉斯细胞组织细胞增生症(LCH)病例进行临床病理回顾性研究及多种免疫表型检测,观察该类疾病的临床病理特征,并对其诊断、鉴别诊断等进行探讨。方法选择原病理诊断为LCH的29例患者为研究对象,分析其临床病理特点;采用链亲和素-生物素-过氧化物酶(SP)法和Elivison二步法检测S-100蛋白、CD1a、CD83及Ki-67在LCH中的表达情况,观察其免疫组织化学结果并进行统计学分析。结果 29例LCH中有5例S-100蛋白、CD1a检测为阴性,排除LCH诊断。在24例LCH中,男性15例,女性9例;患者中位年龄为7.50岁;14例发生于下颌骨,5例发生于上颌骨,5例发生于上下颌骨;按照Bartnick分类,Ⅰ类9例,Ⅱ类13例,Ⅲ类2例;S-100蛋白、CD1a均为阳性表达;颌面部单发骨病损与侵及软组织的颌面部病损相比,Ki-67阳性率较低,而CD83阳性率较高。结论 S-100蛋白、CD1a对于LCH的诊断具有重要意义。颌面部单发的骨LCH可能具有较低的增殖活性,并处于较高的成熟状态;侵及软组织的颌面部LCH可能具有较高的增殖活性,并处于较低的成熟状态。

胚胎小肠S-100^+树突状细胞的组织分布

目的 :观察人胚胎小肠树突状细胞 (dendriticcells,DCs)的组织分布。方法 :应用SABC免疫组织化学法对人胚胎小肠DC的出现时间、分布部位、形态以及数量等进行研究。结果 :( 1 )人胚胎小肠各段S 1 0 0 + DC于第 9～ 1 1w相继出现 ,主要分布于粘膜固有层 ,外形不规则 ,第 2 4w以后 ,其突起相连逐渐在固有层形成网状结构。 ( 2 )回肠集合淋巴小结S 1 0 0 + DC以滤泡间区数量最多 ,其外形相对规则。结论 :人胚胎小肠S 1 0 0 + DC主要分布于固有层 ,其数量随胎龄的增加而逐渐增加 ,而回肠集合淋巴小结S 1 0 0 + DC呈区域性分布 ,它们与固有层S 1 0 0 + DC在形状、大小和数量上有所不同。