Knockdown of FOXM1 suppresses cell growth and metastasis in human laryngeal cancer via the AKT signaling pathway.

OBJECTIVE: This study aims to investigate the potential regulatory effect of forkhead box M1 (FOXM1) on laryngeal carcinoma (LC) and the underlying mechanisms. PATIENTS AND METHODS: Tumor tissues were obtained from 80 patients diagnosed with LC in our hospital. Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot analysis were used to detect the expression levels of FOXM1 in LC tissues and cell lines. Transfection of small interfering RNA (si-RNA) was conducted to knockdown the expression level of FOXM1. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay and cell colony assay were conducted to measure the changes in cell proliferation capacity influenced by FOXM1. Finally, invasion and migration ability was evaluated by the transwell assay. RESULTS: FOXM1 was found upregulated in LC tissues and cells. Transfection of FOXM1 siRNA in LC cells successfully inhibited the expression of FOXM1. The knockdown of FOXM1 resulted in reduced proliferation, invasion, and migration of LC cells. Further studies indicated that the knockdown of FOXM1 suppressed the ratio of p-AKT/AKT. Besides, the impaired proliferation, invasion, and migration of LC cells induced by FOXM1 knockdown could be counteracted by application of the AKT activator Sc79. CONCLUSIONS: The present work demonstrated that the knockdown of FOXM1 suppressed the proliferation, invasion, and migration of LC cells by the AKT signaling pathway.

[Computational investigation of Artemisia pollen deposition in realistic nasal cavities of residents in northwest China].

Objective: To investigate the deposition rate of Artemisia pollen in different nasal cavity regions and its influence factors in residents of northwest China. Methods: Thirty healthy adults from northwest China were enrolled. The computational fluid dynamics (CFD) and discrete phase model (DPM) were used for numerical simulation of nasal structures. The pollen deposition fraction in each anatomical part was counted and the effects of pollen density and breathing rate on deposition were analyzed. SPSS 19.0 software was used for statistical analysis. Results: The hottest deposition parts of Artemisia pollen were nasal septum (30.70%±12.27%), vestibule (27.45%±8.21%), middle turbinate area (13.59%±8.98%) and nasopharynx (7.14%±5.90%). When the inspiratory flow rate increased to 30 L/min, the deposition rates of pollen in nasal vestibule and nasal septum were significantly higher than that at the rate of 15 L/min (43.20%±11.14% vs 27.45%±8.21%, 51.48%±9.77% vs 30.70%±12.27%, t value was -8.126,-5.264, respectively, all P<0.05), which indicated that with the increase of the inspiratory flow rate, the deposition hotspot moved forward. Compared with the wet Artemisia pollen, the deposition rate of the dry pollen in nasal vestibule and nasal septum decreased significantly (16.55%±4.33% vs 27.45%±8.21%, 7.09%±3.69% vs 30.70%±12.27%, t value was 8.669, 9.173, respectively, all P<0.05). The escape rate at outlet increased from 17.00%±9.57% to 43.48%±13.43% (t=-9.282, P<0.05). Conclusions: The deposition of Artemisia pollen in nasal cavity is highly concentrated. The inhalation velocity and the dry-wet degree of pollen are the main determinants of the deposition site.

[Analysis of related factors of recurrent otitis media with effusion in children].

Objective:To investigate the influencing factors of recurrent episodes of otitis media with effusion in children.Method:A retrospective summary of the clinical data of 210 cases of children with otitis media with effusion, 75 cases of recurrence after treatment, 135 cases were recovered, the recurrence of the related factors and after symptomatic treatment effect is analyzed.Result:Logistic regression analysis results found that adenoid hypertrophy (Ⅲ°, Ⅳ°), tonsil hypertrophy (Ⅳ°) and sinusitis (including choanal polyp), a positive allergens, upper respiratory tract infection, the stomach esophagus regurgitation, cleft palate, younger age has significant effect on recurrence of otitis media with effusion, have significant difference (P< 0.05). And the influence of duration, gender, passive smoking history and previous medical history of otitis media with effusion has no obvious statistical significance (P> 0.05). Through the comparison among different age groups, adenoidectomy Ⅲ °, Ⅳ ° hypertrophy tract infections in > 3-6 years old group has significant effect (P< 0.05), recurrent respiratory tract infections in less than 3 years old group and the group of children aged > 3-6 years OME recurrence has significant effect (P< 0.05). By tympanocentesis or tympanostomy tube insertion and according to different conditions to take symptomatic treatment, 75 cases (123 ears) were cured 96 ears (78.05%), 19 ears were improved (15.45%), the total effective rate was 93.50%, ineffective in 8 ears (6.50%).Conclusion:Adenoid hypertrophy (Ⅲ°, Ⅳ°), tonsil hypertrophy (Ⅳ°), sinusitis, nasal polyps, allergic diseases and upper respiratory tract infection gastroesophageal reflux, cleft palate and younger age may be adverse factors related to recurrent otitis media with effusion in children, the clinical doctors should pay attention to these symptoms, according to different causes, adopt individualized treatment plan, make children get the best treatment as soon as possible.

Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)-based scaffolds for tissue engineering.

Development and selection of an ideal scaffold is of importance for tissue engineering. Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) is a biocompatible bioresorbable copolymer that belongs to the polyhydroxyalkanoate family. Because of its good biocompatibility, PHBHHx has been widely used as a cell scaffold for tissue engineering. This review focuses on the utilization of PHBHHx-based scaffolds in tissue engineering. Advances in the preparation, modification, and application of PHBHHx scaffolds are discussed.

Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)-based scaffolds for tissue engineering

Development and selection of an ideal scaffold is of importance for tissue engineering. Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) is a biocompatible bioresorbable copolymer that belongs to the polyhydroxyalkanoate family. Because of its good biocompatibility, PHBHHx has been widely used as a cell scaffold for tissue engineering. This review focuses on the utilization of PHBHHx-based scaffolds in tissue engineering. Advances in the preparation, modification, and application of PHBHHx scaffolds are discussed.

Expression of the genes encoding human leucocyte antigens-A, -B, -DP, -DQ and -G in gastric cancer patients.

This study compared the expression of the genes encoding human leucocyte antigens (HLA)-A, -B, -DP, -DR and -G in peripheral blood mononuclear cells (PBMCs) in gastric cancer patients and healthy controls. Using reverse transcription-polymerase chain reaction, levels of classical HLA-A, -B, -DP and -DR and non-classical HLA-G mRNA were studied in 43 gastric cancer patients and 22 controls. In addition, the levels of HLA-A,B,C and -G antigens on the surface of PBMCs were measured in 30 gastric cancer patients and 15 controls using flow cytometry. The mean fluorescence intensity of HLA-A,B,C antigen in the gastric cancer group was significantly lower than in controls. The HLA-G antigen was mainly present on CD4(+)CD8(-) T-lymphocytes. The percentage of CD4(+)CD8(-) T-lymphocytes positive for HLA-G antigen was significantly lower in the gastric cancer group compared with the healthy controls. Levels of HLA-A, -B and -G mRNA in the gastric cancer group were significantly lower than in controls. The HLA-G mRNA levels were significantly lower in gastric cancer of histological grades III and IV than in grades I and II. These data may provide a novel diagnostic and research tool for gastric cancer.

Association of human papillomavirus type 16 E7 and HLA class I antigen expression in cervical premalignant and malignant lesions.

In the present experiment we studied the correlation between HPV16 infection and expression of HLA-I antigen in cervical premalignant and malignant lesions (cervicitis, CIN, cervical squamous carcinomas and adenocarcinoma samples). The HPV16 E7 DNA load and the expression of HLA-I antigen in the samples were measured by real-time fluorescence quantitative polymerase chain reaction (RFQ-PCR) and immunohistochemical S-P staining, respectively. Our data indicate that HPV16 E7 load was highly and positively associated with the development of cervical lesions (Spearman's correlation coefficient r=0.848, p<0.001), the negative rate of HLA-I antigen was significantly distinguished among groups (p<0.001), and HPV16 E7 infection and downregulation of HLA-I antigen were highly correlated in cervical lesions (Pearson's correlation coefficient r=-0.487, p<0.001). HPV16 E7 may play an important role in the downregulation of HLA-I antigen in cervical lesions, which results in the immune escape of the virus and the occurrence, development, invasion and metastasis of cancer. Furthermore, quantitative PCR for HPV16 E7 may play an important role in the early detection of cervical diseases and in guiding future therapy toward prevention.

Association between polymorphisms of vitamin D receptor gene ApaI, BsmI and TaqI and muscular strength in young Chinese women.

UNLABELLED: The association between the polymorphisms of vitamin D receptor gene ApaI, BsmI and TaqI and isokinetic muscular strength was examined in 109 healthy, young Chinese women. METHODS: Genomic DNA was extracted from mouthwash samples and vitamin D receptor genotypes were determined with the established methods. The peak torque of the dominant limb in concentric and eccentric knee extensors and flexors and elbow extensors and flexors was measured using Cybex-Norm-770 isokinetic dynamometer at slow and fast velocities. The distribution frequency of vitamin D receptor gene alleles was analyzed by chi-square test. The difference in muscular strength between VDR genotype groups was analyzed using one-way ANOVA. The ApaI, BsmI and TaqI alleles and genotype frequencies appeared to be in a Hardy-Weinberg equilibrium. Subjects with the vitamin D receptor ApaI null (A allele) mutation (AA) exhibited significantly lower knee and elbow concentric or eccentric peak torque than those with aa homozygous or Aa heterozygous. The BB & Bb group showed significantly higher peak torque in concentric knee flexors than bb group. No association was found between vitamin D receptor gene TaqI and muscular strength. The data indicated that vitamin D receptor gene ApaI and BsmI polymorphisms rather than TaqI polymorphism might be associated with muscular strength.

Establishment of serum protein pattern for screening colorectal cancer using SELDI-TOF-MS.

AIM: The purpose of this study is to develop a proteomic pattern for distinguishing individuals with colorectal cancer from healthy controls and monitoring micrometastasis using SELDI-TOF-MS. METHODS: A training set consisting of 63 patients with colorectal cancer, 20 patients with benign colorectal diseases and 26 healthy volunteers was used to develop a proteomic model that discriminated colorectal cancer effectively. The sensitivity and specificity of this model was validated by an independent test set. To explore serum proteins changed after operation, the protein profiles of 31 postoperative patients were compared with those of preoperative patients. We also analyzed protein profiles of patients with and without metastasis to monitor micrometastasis. RESULTS: Our study yielded a four-peak model (m/z: 3191.5, 3262.9, 3396.3 and 5334.4) that discriminated cancer from non-cancer samples with sensitivity of 90.3% and specificity of 95.7%. This model was validated in the test set with sensitivity of 87.5% and specificity of 93.8% which was significantly better than the combination use of CEA, CA199 and CA242 (sensitivity 62.4%) for early detection of colorectal cancer. Two peaks (m/z: 2753.8 and 4172.4) were found down-regulated in postoperative samples comparing with preoperative samples. We also detected two proteins (m/z: 9184.4 and 9340.9) that can discriminate patients with primary colorectal cancer from metastatic colorectal cancer. CONCLUSIONS: The four-peak model and two peaks (m/z: 2753.8 and 4172.4) detected in this study have the potential for assistance in diagnostics and therapeutic strategies in colorectal cancer and the two proteins (m/z: 9184.4 and 9340.9) were effective biomarkers for monitoring micrometastasis.

On the deletion of inverted repeated DNA in Escherichia coli: effects of length, thermal stability, and cruciform formation in vivo.

We have studied the deletion of inverted repeats cloned into the EcoRI site within the CAT gene of plasmid pBR325. A cloned inverted repeat constitutes a palindrome that includes both EcoRI sites flanking the insert. In addition, the two EcoRI sites represent direct repeats flanking a region of palindromic symmetry. A current model for deletion between direct repeats involves the formation of DNA secondary structure which may stabilize the misalignment between the direct repeats during DNA replication. Our results are consistent with this model. We have analyzed deletion frequencies for several series of inverted repeats, ranging from 42 to 106 bp, that were designed to form cruciforms at low temperatures and at low superhelical densities. We demonstrate that length, thermal stability of base pairing in the hairpin stem, and ease of cruciform formation affect the frequency of deletion. In general, longer palindromes are less stable than shorter ones. The deletion frequency may be dependent on the thermal stability of base pairing involving approximately 16-20 bp from the base of the hairpin stem. The formation of cruciforms in vivo leads to a significant increase in the deletion frequency. A kinetic model is presented to describe the relationship between the physical-chemical properties of DNA structure and the deletion of inverted repeats in living cells.

Torsionally tuned cruciform and Z-DNA probes for measuring unrestrained supercoiling at specific sites in DNA of living cells.

We describe the development and application of "torsionally tuned" Z-DNA and cruciform probes for analyzing the level of unrestrained supercoiling at specific sites in the DNA of living cells. This approach is applicable for the analysis of dynamic differences in supercoiled DNA in different parts of plasmid, bacterial, or eukaryotic chromosomes. Using a psoralen-based assay, we have shown that the Z-DNA forming sequence (CG)6TA(CG)6, cloned into plasmid pUC8, exists as Z-DNA in 30 to 40% of plasmid molecules in wild-type Escherichia coli. This level suggested an in vivo superhelical density of sigma = -0.034 at the site of insertion in the plasmid. A higher level of Z-DNA found in cells deficient in topoisomerase I (topA10) suggested an in vivo superhelical density of sigma = -0.048. We have constructed a set of torsionally tuned inverted repeated DNA molecules which require different superhelical densities for cruciform formation. Using these inverted repeats and a crosslink assay for cruciforms, we present quantitative evidence for the existence of cruciforms in living E. coli cells. Cruciform formation was dependent on DNA supercoiling in vivo and on the location of the inverted repeat within a plasmid. In topA10 cells cruciforms were detected in less than 0.5% of plasmids when cloned into two different transcriptional units: the lacZ and CAT genes. However, when cloned outside a transcriptional unit, cruciforms were found at levels up to 50% in topA10 cells. More cruciforms were found upstream than downstream from divergent promoters in pBR322. From analysis of the fraction of different inverted repeats existing as cruciforms in vivo and the levels of supercoiling required for cruciform formation in vitro, we estimate in vivo superhelical densities of sigma = -0.034 and -0.041 for the EcoRI site of pUC8-based plasmids in wild-type and topA10 cells, respectively.

Effect of base composition at the center of inverted repeated DNA sequences on cruciform transitions in DNA.

We have analyzed the effect of base composition at the center of symmetry of inverted repeated DNA sequences on cruciform transitions in supercoiled DNA. For this we have constructed two series of palindromic DNA sequences: one set with differing center and one set with differing center and arm sequences. The F series consists of two 96-base pair perfect inverted repeats which are identical except for the central 10 base pairs which consist of pure AT or GC base pairs. The S series was constructed such that the overall base composition of the inverted repeats was identical but in which the positioning of blocks of AT- and GC-rich sequences varied. The rate of cruciform formation for the inverted repeats in plasmid pUC8 was dramatically influenced by the 8-10 base pairs at the center of the inverted repeat. Inverted repeats with 8-10 AT base pairs in the center were kinetically much more active in cruciform formation than inverted repeats with 8-10 GC base pairs in the center. These experiments show a dominant influence of the center sequences of inverted repeats on the rate of cruciform formation.