Objective To investigate the expressions,roles,and clinical significance of microRNA-365(miR-365)and E74-like factor 4(ELF4)in cervical cancer. Methods The expressions of miR-365 in normal cervical tissues(n=34),cervical intraepithelial neoplasia 1(CIN 1)(n=31),cervical intraepithelial neoplasia2-3(CIN 2-3)(n=37),squamous cell carcinoma of the cervix(SCC)(n=33),and three cervical cancer cell lines(C33A cells,Hela cells,and SiHa cells)were detected by real-time quantitative polymerase chain reaction(qPCR).Bioinformatic prediction and luciferase reporter gene assay were performed to verify whether ELF4 was a direct target of miR-365.Western blot and immunohistochemistry were used to detect ELF4 expression in cervical cancer cells and in different pathological cervix tissues.CCK8 assay was used to detect the effect of overexpression or inhibition of miR-365 on the proliferation of cervical cancer cells at different time points.The relationships among the miR-365 expression,ELF4 expression,and clinicopathological parameters of cervical cancer were analyzed by correlation analysis. Results qPCR results showed that compared with the normal cervical cell HcerEpic,the expressions of miR-365 in CIN1,CIN2-3,and cervical cancer tissues gradually decreased with the increased pathologic grade,and its expressions also decreased in different cervical cancer cell lines.The luciferase reporter gene assay confirmed that ELF4 was the direct target of miR-365.Western blot showed that the expression of ELF4 increased in all three cervical cancer cell lines compared with normal cervical epidermal cell(P=0.013,P=0.002,P=0.004).Immunohistochemistry showed that ELF4 expression was up-regulated in CIN and cervical cancer tissues.CCK8 assay showed that overexpression of miR-365 inhibited cell proliferation,while inhibition of miR-365 promoted the proliferation of three cervical cancer cells(P<0.05).Further analysis confirmed that there was a negative correlation between the expression levels of miR-365 and ELF4 in CIN2-3 and SCC(r=-0.351,P=0.045;r=-0.349,P=0.035).Clinical analysis showed that the expressions of both miR-365 and ELF4 were correlated with tumor size,pathological grade,and clinical stage in SCC(all P < 0.05).Conclusion The decreased expression of miR-365 in human cervical cancer cells relieves its inhibitory effect on ELF4,which promotes the proliferation of cervical cancer cells and the formation of tumor.
Cell Proliferation ; DNA-Binding Proteins ; genetics ; Female ; HeLa Cells ; Humans ; MicroRNAs ; genetics ; Transcription Factors ; genetics ; Uterine Cervical Neoplasms ; genetics

As a rare endangered medical plant that newly cultivated,little experimental information is available for growth and metabolites of Tetrastigma hemsleyanum in response to nitrogen( N). The effects of different levels of N on growth of T. hemsleyanum and the content of phytochemicals( polysaccharide,total flavonoids and phenolics) and antioxidant activity( ABTS and FRAP) in stems and leaves were investigated in this study. A certain amount of N had positive effects on most of biological traits,and excessive dose of N went against growth of T. hemsleyanum. With N levels decreased,the polysaccharide content in stems and leaves had no significant change,while the total flavonoid and phenolic content,and antioxidant activities increased steadily. Antioxidant activities and total flavonoid and phenolic content had significant positive correlation. Excessive N fertilizer should be avoided by cultivation.
Antioxidants ; Flavonoids ; Nitrogen ; Phenols ; Phytochemicals ; Plant Extracts ; Plant Leaves ; Vitaceae

BACKGROUND: Uncertainty of repairing articular cartilage defects is highly associated with the mechanical behaviors of the defected area, and the mechanical environment varies with the defect shape, depth and load. OBJECTIVE: To study the mechanical behaviors of articular cartilage defects under physiological load by finite element analysis. METHODS: The axisymmetric model of articular cartilage injury and repair based on transversely isotropy was established using ABAQUS software. The mechanical behaviors of the defect zone repaired with different repair shapes (cylindrical, frustum of a cone, orthorhombic prism, elliptical column) and depths of tissue-engineered cartilage under compressive load were analyzed. RESULTS AND CONCLUSION: The simulation results showed that there were significant differences in the mechanical behaviors of the defect area repaired with tissue-engineered cartilage in different shapes and depths. The stress concentration was the most obvious at the middle-layer defect repair, and the stress distribution was more reasonable at the deep (whole) layer defect repair. Furthermore, the distribution of the stress field and the liquid flow field at the cylinder-shaped tissue-engineered cartilage repair was the closest to the normal cartilage. That is to say, the tissue-engineered cartilage in cylinder or frustum-cone shape is recommended to repair cartilage defect. Importantly, the middle-layer repair is inadvisable.

BackgroundSeveral studies have investigated the association between smoking and anal abscess and anal fistula (AA/F) diseases. However, the relationship between cigarette smoking and AA/F remains unclear. This study sought to assess the role of smoking in anorectal male patients in a Chinese population.

MethodsIn this retrospective study, a questionnaire, including smoking history, was completed over a 3-month period by male inpatients in the Proctology Department of China-Japan Friendship Hospital. "Cases" were patients who had AA/F, and "controls" were patients with other anorectal complaints. Mann-Whitney U-test and Chi-square test were carried out to examine differences in baseline characteristics between groups. Subsequently, multivariate logistic regression was used to explore any related factors.

ResultsA total of 977 patients aged from 18 to 80 years were included, excluding those diagnosed with inflammatory bowel disease or diabetes mellitus. Out of this total, 805 patients (82.4%) completed the entire questionnaire. Among the 805 patients, 334 (41.5%) were cases and 471 (58.5%) were controls. Results showed significant differences between cases and controls (χ = 205.2, P < 0.001), with smoking found to be associated with the development of AA/F diseases (odds ratio: 12.331, 95% confidence interval: 8.364-18.179, P < 0.001).

ConclusionsThis study suggested smoking to be a potential risk factor for the development of AA/F diseases in a Chinese population. Consequently, current smoking patients should be informed of this relationship, and further research should be conducted to explore and investigate this further.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anus Diseases ; epidemiology ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Odds Ratio ; Rectal Fistula ; epidemiology ; Retrospective Studies ; Risk Factors ; Smoking ; adverse effects ; Surveys and Questionnaires ; Young Adult

BACKGROUNDDiabetic nephropathy (DN) is the most common and serious microvascular complication of diabetes. To date, the gold standard for identifying DN and nondiabetic renal disease (NDRD) is a renal biopsy; however, there is currently no reliable diagnostic marker to identify DN and NDRD in a noninvasive manner. This study aimed to investigate the different glycopatterns in urine specimens of DN patients and NDRD patients for a differential diagnosis.

METHODSIn total, 19 DN patients and 18 NDRD patients who underwent renal biopsies between March 2015 and March 2016 at the Chinese People's Liberation Army General Hospital were enrolled in this study. A lectin microarray was used to investigate the glycopatterns in the urinary protein of the 37 patients. Ratio analysis and one-way analysis of variance were used to screen altered glycopatterns. Then, the altered glycopatterns between the DN and NDRD groups were verified by a urinary protein microarray among another 32 patients (15 with DN and 17 with NDRD), and receiver operating characteristic (ROC) curve analysis was used to determine the diagnostic value of the altered glycopatterns in differentiating DN and NDRD. Finally, lectin blotting was used to evaluate the altered glycosylation in protein level.

RESULTSThe result of lectin microarrays revealed that the relative abundance of the (β-1,4)-linked N-acetyl-D-glucosamine (GlcNAc) recognized by lectin Datura stramonium agglutinin (DSA) was significantly higher in urinary protein in DN patients than that in NDRD patients (fold change >1.50, P < 0.001). Subsequently, the results of urinary protein microarrays were consistent with lectin microarrays (P < 0.05). Furthermore, the ROC curve showed that glycopatterns could effectively distinguish DN from NDRD patients (area under the ROC curve = 0.94, P < 0.001). DSA lectin blotting showed that glycoproteins, with a molecular weight of approximately 50,000, demonstrated a difference in urine samples between DN patients and NDRD patients.

CONCLUSIONSThe relative abundance of (β-1,4)-linked GlcNAc recognized by lectin DSA and urinary glycoprotein with a molecular weight of approximately 50,000 are significantly different between DN and NDRD patients, indicating that the glycopatterns could be used as potential biomarkers for a differential diagnosis.

T-cell internal antigen-1 (TIA-1) has roles in regulating alternative pre-mRNA splicing, mRNA translation, and stress granule (SG) formation in human cells. As an evolutionarily conserved response to environmental stress, SGs have been reported in various species. However, SG formation in chicken cells and the role of chicken TIA-1 (cTIA-1) in SG assembly has not been elucidated. In the present study, we cloned cTIA-1 and showed that it facilitates the assembly of canonical SGs in both human and chicken cells. Overexpression of the chicken prion-related domain (cPRD) of cTIA-1 that bore an N-terminal green fluorescent protein (GFP) tag (pntGFP-cPRD) or Flag tag (pFlag-cPRD) induced the production of typical SGs. However, C-terminal GFP-tagged cPRD induced notably large cytoplasmic granules that were devoid of endogenous G3BP1 and remained stable when exposed to cycloheximide, indicating that these were not typical SGs, and that the pntGFP tag influences cPRD localization. Finally, endogenous cTIA-1 was recruited to SGs in chicken cells and tissues under environmental stress. Taken together, our study provide evidence that cTIA-1 has a role in canonical SG formation in chicken cells and tissues. Our results also indicate that cPRD is necessary for SG aggregation.
Chickens ; Clone Cells ; Cycloheximide ; Cytoplasmic Granules ; Humans ; Protein Biosynthesis ; RNA Precursors ; RNA-Binding Proteins ; T-Lymphocytes

OBJECTIVE: To investigate the validity of Coda Motion Analysis System for measuring cervical lateral flexion in normal adults in order to explore a new measuring tool for clinical and research practice. METHODS: A total of 43 participants were involved in the study. Cervical range of lateral flexion were measured with Coda Motion Analysis System and "gold standard" X-ray simultaneously. The validity and agreement were assessed using the scatter diagram, the Pearson correlation coefficient and limits of agreement. RESULTS: Cervical range of lateral flexion measured by Coda Motion Analysis System had no statistical differences with those measured by X-ray(>0.05). The Coda Motion Analysis System demonstrated a very good linear relation with the X-ray measurements in cervical range of right lateral flexion, left flexion and total lateral flexion, and the Pearson correlation coefficients were 0.72, 0.85 and 0.90 respectively. CONCLUSIONS Coda Motion Analysis System showed good validity for measuring cervical lateral flexion in normal adults. Because the reliability of Coda Motion Analysis System was established previously, the results of this study suggest that the system has the potential to be used to measure cervical lateral flexion in clinical and research practice.
Adult ; Cervical Vertebrae ; Eye Diseases, Hereditary ; Humans ; Neck ; Range of Motion, Articular ; Reproducibility of Results

Objective To investigate the feasibility and plan quality of the image-guided volumetric modulated arc therapy (VMAT) based voluntary deep exhale breath-holding technique in the stereotactic ablative body radiotherapy (SABR) for liver tumors.Methods Fifteen patients with liver tumors were involved in this study.All patients were immobilized with voluntary deep exhale breath hold (vDEBH) combined with real-time position management (RPM) respiratory gating system.Treatment was planned using VMAT with 2 modified partial arc and re-planned using intensity modulated radiation therapy (IMRT) technique for comparison.Dosimetric parameters were calculated for plan quality assessment.Quality assurance studies included absolute dose and multiple planar dose verifications,total monitor units and delivery time analysis.Daily cone beam computed tomography imaging was used to verify the motions.Results There were no significant dosimetric differences between VMAT and conventional IMRT plans (P ＞0.05).Both techniques were able to minimize doses to organs at risk including normal liver,kidneys,spinal cord,and stomach.However,the average monitor units with VMAT were significantly lower 28.1％ than those with IMRT(t =3.064,P ＜0.05).The average beam-on time in VMAT plans was 31.6％ shorter than that in IMRT plans(t =2.278,P ＜ 0.05).Conclusions The utilization of VMAT in the treatment planning of SABR for liver tumors under breath control mode has better dosimetrics.In comparison to conventional IMRT plans,VMAT plans have higher efficiency and feasibility.

AIM:To observe the effects of exogenous zinc on the biological behavior of hepatocellular carcino -ma ( HCC) cell line BEL-7404.METHODS: BEL-7404 cells were cultured with zinc sulfate at various concentrations . The intracellular concentration of zinc , cell viability , cell cycle , cell apoptosis and migration and invasion abilities were measured by TSQ fluorescent probe , MTT assay, DNA ploid analysis, acridine orange/ethidium bromide fluorescence stai-ning and Transwell assay , respectively .The mRNA and protein expression levels of albumin in the BEL-7404 cells were determined by real-time PCR and Western blot , respectively .RESULTS: With the elevated concentration of zinc in cul-ture condition, the concentration of zinc in the BEL-7404 cells was increased (P<0.05).The cell viability and migration and invasion abilities were decreased , while the apoptotic rate was increased (P<0.05).The cells in G0/G1 phase were decreased, while the cells in G2/M phase were increased.Additionally, the mRNA and protein expression of albumin also increased (P<0.05).CONCLUSION:The zinc ion inhibits the cell viability as well as migration and invasion abilities , blocks the cells in G 2/M phase , and may reduce cell malignant phenotype .

Objective To study the therapeutic effect of anti-CD80 bivalent antibody on mouse lu-pus nephritis and to explore the possible molecular mechanism. Methods A mouse model of lupus nephritis was established through intraperitoneal injection of 0. 5 ml of pristine in female C57BL/6J mice. Appearance of urinary protein and significantly increased levels of peripheral antinuclear antibody ( ANA) and anti-doub-le-stranded DNA ( anti-dsDNA) antibody in the fourth month after injection indicated that the mouse model was established successfully. Then the mice were divided into two groups including anti-CD80 bivalent anti-body intervention group (injected with 200μg of anti-CD80 bivalent antibody at day 1, 3, 5, 8 and 15, fol-lowed by three times of injection with an interval of one month) and model group ( injected with the same protein using the same strategy). A normal control group was set up accordingly. Albustix test paper was used to monitor the dynamic changes in mouse urinary protein. Flow cytometry was used to analyze the acti-vation of immune-related cells in spleen. Levels of autoantibodies ( ANA and anti-dsDNA) and levels of IFN-γ and IL-4 in serum were detected by indirect immunofluorescence assay. Renal tissue samples were an-alyzed with hematoxylin and eosin ( HE) staining and immunocomplex ( IC) assay. Results Urinary pro-tein level of the anti-CD80 bivalent antibody intervention group was significantly lower than that of the model group (P<0. 05). Activated macrophages, dendritic cells, neutrophils and B cells in spleen tissues of the anti-CD80 bivalent antibody intervention group were significantly less than those of the model group ( P<0. 05), and the numbers of CD4+ and CD154+ T cells were significantly less than those of the model group (P<0. 05). Positive rates and titers of ANA and dsDNA in serum samples of the intervention group were lower than those of the model group (P<0. 05). Levels of IFN-γand IL-4 in serum samples of the interven-tion group were decreased as compared with those of the model group (P<0. 05). HE staining and immuno-fluorescence assay showed that glomerular inflammatory injury and necrosis were alleviated and kidney im-mune complex deposition was reduced after anti-CD80 bivalent antibody intervention. Conclusion Anti-CD80 bivalent antibody specifically binds to the CD80 molecule on antigen presenting cell surface, blocks the CD80/CD28 co-stimulatory signaling pathway and down-regulates the body′s immune response, which al-leviates and reverses the lupus-like nephritis-induced pathological damages in mice.