Objective To explore the changes and significane of USP20 and HIF-α expression in breast cancer.Methods Following transfection of shRNA-USP20 lentivirus into breast cancer MDA-MB-231 cells,the gene and protein expression levels of USP20 were detected using fluorescence quantitative PCR and Western Blot.Subsequently,the overexpression of USP20 was observed to determine its effect on HIF-α expression.Similarly,siRNA-USP20 was used to knock down USP20 in breast cancer MDA-MB-231 cells,followed by detection of gene and protein expression levels using fluorescence quantitative PCR and Western Blot.The subsequent changes in HIF-α expression were then examined.Rusults The positive expression rates of USP20 and HIF-α in breast cancer tissues were 69.6%and 46.83%,respectively,while they were negatively expressed in the adjacent normal tissues,with statistically significant differences(P＜0.01).The positive expressions of USP20 and HIF-α were predomi-nantly observed in the cytoplasm of breast cancer tissue,with a smaller amount present in the nucleus.There was a significant positive correlation between USP20 and HIF-α in breast cancer.Following transfection of shRNA-USP20 lentivirus into MDA-MB-231 cells,both the protein and gene expression levels of USP20 significantly increased(P＜0.01).Over-expression of USP20 did not affect HIF-α mRNA levels but led to a significant increase in HIF-α protein expression(P＜0.01).Conversely,siRNA-USP20 interference resulted in a significant decrease in both the protein and gene expression levels of USP20(P＜0.01),without affecting HIF-α mRNA levels;however,it caused a notable reduction in HIF-α protein expression(P＜0.01).Conclusion The expression of USP20 exhib-ited a significant positive correlation with HIF-α in breast cancer.Overexpression of USP20 led to a substantial increase in HIF-α protein expression,while knock-down of the USP20 gene resulted in a significant decrease in HIF-α protein levels.Therefore,it can be inferred that USP20 may exert its influence on the development of breast cancer through modulation of HIF-α expression,thereby providing crucial experimental evidence for clinical treat-ment,prognosis,and further investigations.

Immunotherapy has recently become a new therapeutic method. Related cells and cytokines mediating tumor immunity play different roles in the process of tumor immunity, and they are correlated with tumor prognosis, which has a reference significance in the evaluation of tumor prognosis. In China, esophageal carcinoma is one of the common tumors of digestive system, and its 5-year overall survival rate is low. With the wide use of immunotherapy and introduction of precision medicine, the study on the relationship between tumor immune-related cells, cytokines and the prognosis of esophageal carcinoma can provide the guidance for the diagnosis and treatment of esophageal carcinoma.

Objective To investigate the effect of ANX A1 on the biological characteristics of papillary thyroid carcinom cells by interfering with the expression of Annexina A1 (ANX A1) in papillary thyroid carcinoma cells through small interfering RNAs (siRNA).Methods The designed highly efficient siRNA was used to conduct the specific interfence on ANXA1 expression in the papillary thyroid carcinoma TPC-1 cells.The effect of ANXA1 on TPC-1 apoptosis in PTC was observed by flow cytometry.Results The designed siRAN could efficiently inhibit the expression of ANXA1 mRNA in PTC,enhanced the cell apoptosis in TPC-1 cells in vitro.Conclusion siRNA can interfere with the expression of ANXA1 and promote the apoptosis of papillary thyroid carcinoma which suggesting that ANXA1 may be an important biological target for the treatment of papillary thyroid carcinoma.

Objective This study is to investigate the influences of USP7 on the cytobiological characteristics of laryngeal cancer cells by small interfering RNA (siRNA) interfering the USP7 expression in the laryngeal cancer cells .Methods The self‐de‐signed highly efficient siRNA was used to conduct the specific interference on USP7 expression in laryngeal cancer HEP2 cells . Then the influence on the capacity of cell proliferation and migration ,as well as apoptosise after USP7 interference were observed by using the CCK‐8 method ,Transwell chamber migration test and flow cytometry .Results The self‐designed siRNA could effi‐ciently inhibit the expression of USP7 mRNA in laryngeal cancer cells ,furthermore markedly suppressed the proliferation and mi‐gration of laryngeal cancer cells ,enhanced the cell apoptosis in laryngeal cancer HEP2 cells in vitro .Conclusion The siRNA inter‐fering USP7 can inhibit the proliferation and migration capacity of laryngeal cancer cells ,and promoted their apoptosis .

Objective To explore the correlations between Annexin A1 protein expression and clinicopathological character-istics in carcinoma of papillary thyroid.Methods The different expressions of annexin A1 in papillary thyroid tissue and para-cari-noma tissue were investigated by immunohistochemistry.Results Among 69 samples tissues of papillary thyriod carcinoma,the positive rate of annexin A1 was higher than that of 69 para-carcinoma tissues(88.41%vs .8.69%),there was a significant difference (P <0.05).Furthermore,the expression of annexin A1 was correlation with the lymph node metastasis and tumor size,which was higher in ≥1 cm diameter of tumor(P <0.05).Conclusion High AnnexinA1 positive expression in papillary thyroid cancer tissues is associated with tumor malignant progression,which might be a valuable predictor and potential target for the diagnosis and treat-ment of papillary thyroid carcinoma.

Objective To study the effect of annexin A5 on the apoptosis of laryngeal cancer cells.Methods Special siRNAs were used to knock annexinA5 down in Hep-2 cell,and RT-PCR and Western blot were applied to identify the efficacy of RNA interference.The flow cytometry assay was performed to detect the Hep-2 cell apoptosis.Results RT-PCR analysis showed that the relative mRNA expression of annexin A5 in siRNA group,negative control group,Lipofectamine2000 group and blank control group were 0.70 ±0.03,1.18 ±0.05,1.17 ±0.06 and 1.23 ±0.07.The relative mRNA expression of annexin A5 in siRNA group was significantly decreased than contrast groups(t =-14.77,t =-13.23,t =-12.99,P ＜0.05).In Western blot assay,the trend of protein expression level was consistent with the mRNA expression levels of annexin A5.The relative levels of proteins in siRNA group,negative control group,Lipofectamine2000 group and blank control group were shown 1.21 ± 0.03,3.88 ± 0.06,3.87 ±0.02 and 3.95 ± 0.08.The relative protein expression of annexin A5 in siRNA group was significantly decreased than contrast groups(t =-70.34,t =-150.62,t =-56.32,P ＜0.05).At the same time in flow cytometry the apoptotic rate of siRNA group,negative control group,Lipofectamine2000 group and blank control group were 4.43％ ±0.12％,13.67％ ±0.22％,13.66％ ±0.12％ and 13.35％ ±0.13％,the difference between the siRNA group and contrast groups was statistically significant (t =-62.50,t =-14.16,t =-11.47,P ＜ 0.05).So after RNA interference,expression of annexin A5 decreased,and the results in the apoptosis inhibition of Hep-2 cell.Conclusion Annexin A5 promotes apoptosis of Hep-2 cells,and it may be a potential therapeutic target for the laryngeal cancer.

Objective To study the relationship between the fbxl 20 gene、E-cadherin gene and the clinocopathologic features ,respectively , and the correlation about the fbxl 20 and the set gene .Methods The mRNA expression level of fbxl 20 and E-cadherin gene in 50 pairs of human colorectal adenocarcinoma tissues and matched normal tissues were detected by RT -PCR.Results The mRNA expression level of fbxl 20 gene in tumor tissues were significantly increased than that in normal tissues (0.479 ±0.141 vs.0.296 ±0.121,P=0.001),while the E-cadherin were decreased (0.440 ±0.026 vs.0.741 ±0.059,P=0.000).There was no found on the correlation between the mRNA expression level of fbxl 20 and the E-cadherin gene in the 50 tumor tissues(r=-0.165,P=0.251).However,the E-cadherin expressed level was negatively correlated with the fbxl 20 expressed level in the lower differentiation degree、in the high differentiation degree and the Duke′s D tumor tissues(r=-0.600,P=0.008;r=-0.784,P=0.017;r=-0.643,P=0.032).Conclusions The decreased mRNA expression level of E -cadherin in tumor tissues are largely due to the increased fbxl 20 gene expressed level ,which is related with the high mobility and invasion ability of the advanced tumor .

Japanese encephalitis virus (JEV) is the leading cause of viral encephalitis in Asia and domestic pigs serve as the amplifying hosts. In the present study, the full genomic sequences of two JEV strains (HEN0701 and SH0601) isolated from pigs in China were determined and compared with other 12 JEV strains deposited in GenBank. These two strains had an 88.8% nucleotide sequence similarity and 97.9% deduced amino acid sequence homology. HEN0701 had high nucleotide sequence and high amino acid sequence identity with genotype I (GI) strains, while SH0601 had high nucleotide sequence and high amino acid sequence identity with GIII strains at both the gene and full genome levels. Further phylogenetic analysis showed that HEN0701 belonged to the JEV GI group and SH0601 was classified as a GIII strain. Analysis of codon usage showed there were a few differences between the GI and GIII strains in nucleotide composition and codon usage for the open reading frames.
Animals ; Cell Line ; Cricetinae ; Encephalitis Virus, Japanese/classification/*genetics ; Encephalitis, Japanese/epidemiology/*veterinary/virology ; Gene Expression Regulation, Viral/physiology ; Genome, Viral ; Molecular Epidemiology ; Phylogeny ; Swine ; Swine Diseases/epidemiology/*virology

Japanese encephalitis virus (JEV) is the leading cause of viral encephalitis in Asia and domestic pigs serve as the amplifying hosts. In the present study, the full genomic sequences of two JEV strains (HEN0701 and SH0601) isolated from pigs in China were determined and compared with other 12 JEV strains deposited in GenBank. These two strains had an 88.8% nucleotide sequence similarity and 97.9% deduced amino acid sequence homology. HEN0701 had high nucleotide sequence and high amino acid sequence identity with genotype I (GI) strains, while SH0601 had high nucleotide sequence and high amino acid sequence identity with GIII strains at both the gene and full genome levels. Further phylogenetic analysis showed that HEN0701 belonged to the JEV GI group and SH0601 was classified as a GIII strain. Analysis of codon usage showed there were a few differences between the GI and GIII strains in nucleotide composition and codon usage for the open reading frames.
Animals ; Cell Line ; Cricetinae ; Encephalitis Virus, Japanese/classification/*genetics ; Encephalitis, Japanese/epidemiology/*veterinary/virology ; Gene Expression Regulation, Viral/physiology ; Genome, Viral ; Molecular Epidemiology ; Phylogeny ; Swine ; Swine Diseases/epidemiology/*virology

Abnormal expression of annexin A2 contributes to metastasis and infiltration of cancer cells.To elucidate the cause of abnormal expression of annexin A2,Western blotting,immunoproteomics and immunohistochemical staining were performed to analyze differentially ubiquitinated proteins between fresh breast cancer tissue and its adjacent normal breast tissue from five female volunteers.We detected an ubiquitinated protein that was up-regulated in the cancer tissue,which was further identified as annexin A2 by mass spectrometry.These results suggest that abnormal ubiquitination and/or degradation of annexin A2 may lead to presence of annexin A2 at high level,which may further promote metastasis and infiltration of the breast cancer cells.