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Objective@#To investigate alteration of proteins profile in malignant transformation bronchial epithelial cells(16HBE-T) induced by hexavalent chromium[(Cr(VI))] and analyze the expression level of SET protein, then to provide some new insights for the carcinogenesis mechanism of Cr(VI).@*Methods@#Total protein was extracted from 16HBE cells and was alkylated and desalinated before digested into peptides. The products were labeled with Tandem Mass Tag (TMT) and identified using LC-ESI-MS/MS.@*Results@#A total of 3 517 proteins were found, expression differences greater than 1.5 or less 0.67 times were to found have 185 and 201 proteins, respectively. Gene enrichment analysis revealed that differential proteins were mainly involved in autophagy, DNA damage repair, RNA processing and other biological processes. Western blot results showed the expression level of SET was significantly increased while downregulated in histone H3K18/27 acetylation and p53 protein.@*Conclusion@#Proteins involved in multiple biological processes altered in 16HBE-T cells and regulation mode of SET inhibiting histone H3K18/27 acetylation regulating transcriptional activity of p53 may paly an important role in Cr(VI)-association carcinogenesis.
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Objective@#To investigate DNA damage in the transformed human bronchial epithelial cells (16HBE) induced by hexavalent chromium (Cr6+) and further elucidate the potential carcinogenesis mechanism of Cr6+.@*Methods@#16HBE were treated with different concentration of Cr6+ (0, 0.625, 1.25, 2.5 μmol/L) for 15 weeks. The malignant degrees of transformed cells were identified by the assays for anchorage-independent growth and tumorigenicity. According to the single cell gel electrophoresis (SCGE) assay, the DNA damage rate was calculated. The expression level of 53BP1 was determined by Western blot.@*Results@#Chromium-treated cells could form colonies in soft agar and tumors in nude mice. Compared with the control group, colony formation efficiency of 1.25μmol/L and 2.5 μmol/L Cr6+-treated cells in soft agar showed significant increases (p<0.05) . The 2.5 μmol/L Cr6+-treated cells also formed tumors subcutaneously in nude mice. Cr6+ could cause different degree of DNA damage to 16HBE cells in a dose-dependent manner. In addition, Western blot analyses showed that 53BP1 was aberrantly down-regulated at 2.5 μmol/L dose and has no significant changes at 0.625 μmol/L and 1.25 μmol/L dose under the treatment of Cr6+.@*Conclusion@#The declined expression of 53BP1 may mediate Cr6+-induced DNA damage and further involved in the cell malignant transformation.
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Since smart phones have been developed, significant advances in the function of mobile phone due to the development of software, hardware and accessories have been reached. Till now, smart phones have been engaged in daily life with an increasing impact. As a new medical model, mobile phone medicine is emerging and has found wide spread applications in medicine, especially in diagnosing, monitoring and screening various diseases. In addition, mo bile phone medical application shows great potential trend to improve healthcare in resource-limited regions due to its advantageous features of portability and information communication capability. Nowadays, the scientific and technological issues related to mobile phone medicine have attracted worldwide attention. In this review, we summarize state-of-the-art advances of mobile phone medicine with focus on its diagnostics applications in order to expand the fields of their applications and promote healthcare informatization.
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Humans , Cell Phone , Delivery of Health Care , SoftwareABSTRACT
ObjectiveTo observe the influence of lipopolysaccharide (LPS) on collagen metabolism of normal human skin fibroblasts and its biological role in the formation of hypertrophic scar.Methods Fibroblasts were isolated and cultured in vitro,and then exposed to different doses of LPS (0.005,0.01,0.05,0.1,0.5,1.0 μg/ml) from E.coli.055:B5 respectively.The expression of proccllagen type Ⅰ,Ⅲand collagenase mRNAs was tested by RT -PCR.Fibroblasts from hypertrophic scar tissue obtained from the same patients in the same culture passage were used as control.ResultsCompared with control group,the expression of procollagen typeⅠ,Ⅲ mRNAs in normal skin fibroblasts increased (0.323 ± 0.041,0.303 ± 0.063,0.391 ± 0.071,0.344 ± 0.086,0.488 ± 0.059,0.401 ± 0.087,0.616 ± 0.107,0.434 ±0.084,0.823 ±0.092,0.542 ± 0.082),while the expression of collagenase mRNAs of normal skin fibroblasts depressed(0.598 ± 0.068,0.556 ± 0.049,0.441 ± 0.043,0.372 ± 0.083,0.260 ± 0.027 ).When LPS was set to the concentration of 0.005 μg/ml,it showed a concentration dependent manner.However,when the concentration of LPS was set to 0.5 μg/ml,the expression of procollagen type Ⅰ,Ⅲ and collagenase mRNAs of normal skin fibroblasts began to decrease (0.451 ± 0.063,0.374 ± 0.072,0.360 ± 0.062).When the concentration of LPS was set to 1.0 μg/ml,the expression of procollagen type Ⅰ,Ⅲ mRNAs (0.162 ± 0.025,0.171 ± 0.061 )were inhibited and the expression of collagenase mRNAs began to increase (0.444 ±0.114).When the concentration of LPS was set to 0.1 μg/ml,the expression of procollagen type Ⅰ,Ⅲ and collagenase mRNAs of normal skin fibroblasts(0.823 ±0.092,0.542 ±0.082,0.260 ±0.027)was similar to that of hypertrophic scar tissue fibroblasts(0.829 ±0.049,0.569 ±0.038,0.277 ±0.059).ConclusionsThis result supported that LPS may be an important factor in collagen metabolism of normal skin fibroblasts and it plays an important role in hypertrophic scar formation.
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Objective To observe the influence of lipopolysaccharide (LPS) on the cell cycle and the mRNAs expression of procollagen type Ⅰ , Ⅲ of normal human skin fibroblasts. Methods Purified dermal fibroblasts were exposed to different doses of LPS(0. 005 ～ 1.0 μg/ml) from E. coli. Then the cell cycle of fibroblasts at logarithmic stage at day 7 after LPS administration was assayed with flow cytometry.The expression of procollagen type Ⅰ , Ⅲ and collagenase mRNAs was tested by RT-PCR. Results The percentage of S phase cells in cell cycle of normal human skin fibroblasts increased when LPS concentrations were changed from 0. 005 to 0. 1 μg/ml, and the increase showed a concentration dependent manner. However, when the concentration of LPS was 0. 5 μg/ml, the percentage of S phase cells began to decrease, but still higher than normal control. When LPS concentration reached 1.0 μg/ml, the percentage of S phase cells were lower than normal control. The expression of procollagen type Ⅰ , Ⅲ mRNAs of normal skin fibroblasts increased when LPS was challenged to the concentration of 0. 005 μg/ml, and the influence showed a concentration dependent manner. However, when the concentration of LPS was 0. 5 μg/ml, the influence of LPS on the expression of procollagen type Ⅰ , Ⅲ of normal skin fibroblasts began to decrease.When the concentration of LPS reached 1.0 μg/ml, the expression of procollagen type Ⅰ , Ⅲ mRNAs were inhibited. Conclusions LPS promoted the proliferation and collagen synthesis of normal human skin fibroblasts within a certain range of low doses, while high doses of LPS might inhibit the proliferation and collagen synthesis of normal human skin fibroblasts.
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Objective To explore the relationships among pulmonary function,DVH and acute radiation pneumonitis after three-dimensional conformal radiation treatment in patients with non-small-cell lung cancer.MethodsPulmonary function tests were conducted on 68 inoperable patients (male 42,female 26,median age 52,KPS≥80) before and after three months radiotherapy respectively.After 3 months of follow-up,radiation pneumonitis were graded,and V20,V30 and MLD were derived from dose volume histogram (DVH).ResultsAll patients were treated with radiotherapy at the irradiation dose of 60～70Gy.Acute radiation pneumonitis occurred in 24 patients with 11 Grade Ⅰ,7 Grade Ⅱ,3 Grade Ⅲ,3 Grade Ⅳ.There were no significant difference between the pre-irradiation and the three months after irradiation for FVC (P＞0.05).But there were significant different between pre-irradiation and three months after irradiation for FEV1.0 and DLCO (P＜0.05).V20,V30 and MLD were observed in patients treated with high radiation pneumonitis.ConclusionsThere were close relationships among pulmonary function,DVH and radiation pneumonitis in patients with non-small cell lung cancer.
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Objective To study LI gene sequence of HPV cp6108 from 5 cases of condyloma acuminata. Methods T-A cloning and direct sequencing of PCR product were used. Results The LI gene sequences of HPV cp6108 from 5 specimens were presented with the homology of 99% to reference sequence in GenBank. A total of 3 gene mutations were found, including a nonsense mutation of G70A, a missense mutation of D77N, and a missense mutation of Tl16P. Conclusions In comparison with the sequence in GenBank, at least 3 gene mutations of HPV CP6108, i.e. one nonsense mutation of G70A and missense mutations of D77N and Tl 16P, are found in the present study.
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Objective To screen primers used in polymerase chain reaction (PCR) for detecting Trichomonas vaginalis. Methods Three pairs of PCR primer reported in the literatures (TVA5-TVA6, TV1-TV2 and TVK3-TVK7) were screened. For each PCR, four components, including primers, Mg2+, dNTPs and Taq polymerase, were optimized using Taguchi methods to determine the optimal PCR conditions. With the optimal conditions, the sensitivities of three PCR were compared. Vaginal swabs were collected to detect Trichomonas vaginalis by culture and PCR, and the PCR with highest sensitivity was evaluated. Results All three PCR were of high specificity, and the PCR with primers of TVK3-TVK7 had the highest sensitivity. Of 25 clinical vaginal swabs, T. vaginalis was detected in 7 samples by the culture, however, it was detected in 8 samples by the PCR. All culture-positive samples were also positive by PCR. Conclusions The PCR with the primers of TVK3-TVK7 is highly sensitive and specific, which could be useful to detect T. vaginalis in vaginal swab samples.
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Objective To systematically investigate the molecular epidemiological profiles of human papillomavirus (HPV) in patients with condyloma acuminata(CA). Methods Two hundred and one samples of HPV DNA isolated from CA were PCR amplified by the PGMY09/11 primer system. The PCR products were simultaneously hybridized to 37 specific HPV probes immobilized on a nylon strip and then genotyped. All DNA templates were further PCR amplified using HPV 6 and 11 type specific primers for verification. Results All samples were HPV DNA positive consisting of totally 31 genotypes, the types of which were type 11(53.7%, 108/201), 6(43.8%, 88/201), 16(6.5%, 13/201), 52(6.0%, 12/201), 33(5.5%, 11/201), cp6108 (5.5%, 11/201) and 42 (5.0%, 10/201). The samples infected with a single and mixed types of HPV accounted for 60.2% (121/201) and 39.8% (80/201) respectively. Consistent results were found with the detection of HPV6 and 11 between hybridization assay and type-specific PCR. Conclusions At least 31 HPV genotypes are associated with CA. HPV 11 predominates while 68, 40, 54, 67, 73, 82, 35, 64 and 83 are rare in CA. Type cp6108 is detected in CA for the first time with a high prevalence. HPV26, 69, 70, 71,72 and IS39 might be not associated with CA. CA infected with a single and mixed HPV types accounts for 60.2% and 39.8%, respectively.
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Objective To clone and express immunodominant fragment of glycoprotein G of HSV-2 (FgG-2). Methods The target gene was amplified by polymerase chain reaction (PCR). The PCR products were ligated into directional TOPO expression vector. After identification, the recombinant expression vector was transferred into BL21 StarTM cell for expression. Finally, recombinant protein of FgG-2 (rFgG-2) was detected by Western Blot (WB). Results A 616 bp DNA fragment was obtained with PCR and then confirmed in recombinant vector by PCR and sequencing, bearing 99.5% consistent sequence with target gene. Highest recombinant protein production was obtained at the time point of 3 hours. Expression of target protein was confirmed by WB with anti-gG monoclonal antibody. Conclusions The immunodominant fragment of gG-2 has been successfully cloned and expressed in E.coli, which might be used for the development of serum diagnostics assay kits for HSV-2 infection.
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Objective To develop a gene chip for the detection of pathogens causing genital ulcer diseases (GUDs). Methods Specific probes of 4 different pathogens were designed and synthesized. Gene chip was prepared by blotting the probes onto specially treated glass slides with the use of a robotics. Target genes of standard strains for the 4 different pathogens and the clinical specimens were amplified by PCR with Cy5 fluorescence labeled primers. The labeled amplicons were hybridized with gene chips, and then scanned and analyzed using computer software. Results The fluorescence signal for specific pathogen could be found in the gene chip, illustrating that one specific fluorescence signal denoted a single pathogen, and the combination of different signals denoted the corresponding co-existence of pathogens. Examination of 40 clinical specimens obtained from 40 patients with genital ulcers with gene chip was in good concordance with dark-field microscopy plus PCR or HSV culture plus PCR, showing Kappa values of 0.882 and 0.947, respectively. In addition, mixed infections were detected in 2 specimens. Conclusion Gene chip is a sensitive method with a reliable result and it can detect multiple infections simultaneously.