Objective To establish a method for detecting the EGFR gene mutations by the real-time fluorescence quantification PCR combined with Sanger sequencing and to preliminarily explore its clinical application value .Methods With EGFR gene hotspot mutations region exon 19 and 21 as the research locus ,the specific amplification and the sequencing primer were designed ,the known wild-type and mutant samples were utilized to construct the corresponding plasmid as the standard substance by the TA clone technique .Then the EGFR gene mutation detection method by the real-time fluorescence quantification PCR combined with Sanger sequencing was established and the methodological and the application evaluation were performed .Results The wild-type and mutant standard plasmids of the EGFR gene exon 19 and 21 were constructed successfully .The EGFR gene mutations detection method of the real-time fluorescence quantification PCR combined with Sanger sequencing was established ,which had high sensitivi-ty(101copies/μL)andgoodrepeatability(intra-assayCVandinter-assayCVofthereal-timefluorescencequantificationPCRofex-on 19 and 21 were 1 .42% /3 .52% and 0 .97% /2 .44% ,respectively ) .20 clinical samples were simultaneously detected by this method and the traditional Sanger sequencing ,the results were completely consistent .Conclusion The EGFR gene mutations detec-tion method of the real-time fluorescence quantification PCR combined with Sanger sequencing is successfully established ,which can be used in the clinical sample detection .

Health Knowledge, Attitudes, Practice ; Humans ; Occupational Diseases ; prevention & control ; Rural Population ; Sampling Studies ; Surveys and Questionnaires ; Transients and Migrants

An electrospray/ultraviolet lamp dual-source ion trap mass spectrometer was developed for the rapid detection of gas and liquid samples.The instrument used the discontinuous atmospheric pressure sampling technique that both the electrospray ions and gaseous analytes were sampled and transferred using a pinch valve device.The two ionization sources were generally suitable for different kinds of analytes and provided complementary applications.Electrospray was used for the ionization of polar compounds in solution, while the UV ionization source was mainly applied for the analysis of gaseous organic compounds.A variety of samples such as anisole, toluene, 2,4-dimethylaniline, arginine, reserpine and aspartame were employed to test the performance in different working mode of the instrument.The results showed that the two sources were feasible for ionization of different types of samples, and different types of molecular ions could be generated when 2,4-dimethylaniline was analyzed.The two ionization sources could be used alternately or simultaneously without interference, and the working mode was also switched to fit the application requirements.The dual-source configuration was an effective way to extend the applications for miniature mass spectrometers.It did not significantly increase the size of the instrument, but provided more versatile analysis to meet the need for the measurement of different types of samples.

Objective Bladder cancer , which has a high rate of recurrence and invasion , is the most common genitourinary cancer.The article was to study the effect of specific chemokine receptor CXCR 4 on invasion capacity and intraluminal implantation of human bladder cancer cells . Methods A CXCR4 specific recombinant plasmid vector (short hairpin, shRNA) was constructed to select those cells which could inhibit the expression of CXCR 4, and these cells were divided into blank control group , negative control plasmid group and recombinant plasmid group (pshRNA-CXCR4-1, pshRNA-CXCR4-2).RT-PCR and immunofluorescence technique were used to detect the mRNA and protein expression of CXCR 4 respectively .Invasion capability in vitro of the cells was evaluated by Boyden chamber .20 nude mice were randomly divided into experimental group and control group ( n=10 ) .The experimental group was established by injection of 100μL shRNA-EJ-M3 into the bladder , while the control group was established by injection of 100μL EJ-M3, aiming to detect the effect of shRNA-CXCR4 on intraluminal implantation of human bladder cancer cells . Results The CXCR4 mRNA expression of the pshRNA-CXCR4-1 group (62.05 ± 1.35) was significantly lower than that of blank control group (174.38 ±1.96, P <0.05 ) and negative control plasmid group (166.27 ±1.82, P <0.05).There was no significant difference among the CXCR4 mRNA expression of the pshRNA-CXCR4-2 group, blank control group and negative control plasmid group ( 182.58 ± 4.2, 174.38 ±1.96, 166.27 ±1.82, P>0.05).In immunofluores-cence experiment, the red cell amount of the pshRNA-CXCR4-1 group(32.24 ±2.23) was lower than that of the blank control group (89.61 ±4.47,P<0.05) and negative control plasmid group (92.45 ±3.68, P<0.05).There was no significant difference among the red cell amount of the pshRNA-CXCR4-2 group, the blank control group and the negative control plasmid group (76.87 ±5.11, 89.61 ±4.47, 92.45 ±3.68, P>0.05).The Boyden chamber experiment showed that the number of penetrating cells of the pshRNA -CXCR4-1 group (39.67 ±8.45) was significantly lower than that of the blank control group (135.33 ±9.28, P<0.05) and that of the negative control plasmid group(123.63 ±6.36, P<0.05).As to the intraluminal implanting capability, the difference between the ex-perimental group and the control group of statistical significance (10%vs 70%,P<0.01). Conclusion CXCR4 shRNA can inhibit the expression of CXCR4 and significantly decrease the invasion capacity and intraluminal implantation of human bladder cancer cells .

[ ABSTRACT] AIM: To observe the effects of TNF-α/nuclear factor-κB( NF-κB) /matrix metalloproteinase-2 (MMP-2) pathway on the expression of MMP-2 in the mice with viral myocarditis.METHODS: Six-week-old inbred male mice were randomly assigned to control and myocarditis group.The mice in myocarditis group and control group were intra-peritoneally inoculated with 0.1 mL 10-5.69 TCID50 /mL coxsackievirus B3 and vehicle (PBS), respectively.Ten mice were sacrificed at the 4th and 10th days after injection.The blood and heart specimens were harvested.The serum content of TNF-αwas measured by ELISA.The myocardial levels of MMP-2, NF-κB p65 and IκBαwere determined by Western blot.RESULTS: Compared with control group, the protein expression of MMP-2 and NF-κB p65 in the myocardium and the serum content of TNF-αwere significantly increased in myocarditis group (P <0.05).The protein expression of IκBαwas lower in myocarditis group than that in control group (P <0.05).CONCLUSION: TNF-α, NF-κB p65 and MMP-2 were higher in the mice with acute viral myocarditis.The increased expression of them might be involved in the pathogene-sis of viral myocarditis.

The study of viruses traditionally focused on their roles as infectious agents and as tools for understanding cell biology. Recently, however, with the development of structural biology, viruses have now been receiving particular attention in nanotechnology. By chemical methods or by gene modification, viruses have been functionalized as potential building blocks for several applications, such as drug/gene delivery vehicles, advanced vaccine vehicles, and special inorganic or organic nanomaterials. Here we highlight some of the recent progresses in the medical applications of viruses.
Biomedical Research ; trends ; Gene Transfer Techniques ; Genetic Therapy ; Nanostructures ; Nanotechnology ; trends ; Vaccines ; Viruses

Female ; Humans ; Infant, Newborn ; Maple Syrup Urine Disease ; diagnosis ; therapy

Objective To investigate the predictive factors of progressive motor deficits (PMD) after isolated pontine infarction. Methods Consecutive patients with isolated pontine infarction admitted to hospital within 48 hours after onset were enroled. They were divided into either a PMD group (increase ≥1 within 7 days) or a non-PMD group according to the clinical course and the changes of motor scores of the National Institutes of Health Stroke Scale (NIHSS). The pontine infarction patterns were classified as basal surface infarction and deep infarction, the sides were divided into left and right, the infarct levels were divided into upper, middle, and lower according to diffusion-weighted imaging. The demographics, baseline clinical data, and imaging features were compared between the two groups. Multivariable logistic regression models were used to analyze the predictive factors of PMD after isolated pontine infarction. Results A total of 101 patients with isolated pontine infarction admitted to hospital within 48 h of onset were enroled, including 16 in the PMD group and 85 in the non-PMD group. The proportions of pontine infarction involving the basal surface (87. 5% vs. 47. 1% , χ2 = 8. 851, P = 0. 003), the infarcts on the middle levels (56. 2% vs. 24. 7% , χ2 = 4. 851, P = 0. 028), and basilar artery stenosis or occlusion (62. 5% vs. 27. 1% ,χ2 = 7. 689, P = 0. 006) of the PMD group were significantly higher than those of the non-PMD group, while the proportions of the infarcts on the left sides (18. 8% vs. 56. 5% , χ2 = 7. 664, P = 0. 006) and the infarcts on the upper levels (37. 5% vs. 72. 9% , χ2 = 7. 689, P = 0. 006) of the PMD group was significantly lower than those of the non-PMD group. Multivariate logistic regression analysis identified that pontine infarction involving the basal surface (odds ratio 5. 650, 95% confidence interval 1. 011 - 31. 580, P = 0. 049) and basilar artery stenosis or occlusion (odds ratio 4. 075, 95% confidence interval 1. 127 - 14. 741, P = 0. 032) were the independent risk factors for PMD after isolated pontine infarction. Conclusions Infarction involving the basal surface and basilar artery stenosis or occlusion may be the predictors for PMD after isolated pontine infarction.

Objective To evaluate the olfactory functions in patients with Parkinson's disease (PD) with a smell identification test developed for the Chinese population and to examine possible factors contributing to PD patients' olfactory performance.Methods Fifty-five patients with idiopathic and nondementia PD and 55 demographically matched healthy controls (HC) were recruited and their olfactory identification tested using the Chinese Smell Identification Test developed by the Institute of Psychology at Chinese Academy of Sciences.Results Olfactory identification score was significantly lower (t =-6.827,P＜0.01) in the PD group (19.3 ±5.4) as compared with the HC group (26.5 ±5.7).Of all PD patients,63.6％ scored over 1 SD below the age norm.In particular,of those with Hoehn-Yahr ratings between 2 to 3,77.8％ scored over 1 SD below the age norm.Receiver operating characteristic (ROC) curve analysis indicated that a cut-off score of 22.5 on the Chinese Smell Identification Test distinguished between PD patients and HC with a sensitivity of 74.6％ and specificity of 70.9％.Within the PD group,age,education,Hoehn-Yahr rating and Mini-Mental State Examination score were not found to correlate with olfactory performance (all P ＞ 0.05).Conclusion PD patients in China show significant impairment of olfactory identification,which is effectively captured by the Chinese Smell Identification Test.

Objective To analyze the perioperative clinical characteristics of non-small cell lung cancer ( NSCLC ) complicated with dermatomyositis ( DM) . Methods Six cases of NSCLC complicated with DM received lobectomy and lymph node dissection by video-assisted thoracoscopic surgery. The same kind of operations were carried out in another six NSCLC cases without DM in the same period. The twelve cases in two groups were 1 ∶ 1 matched study. The operation time, operative blood loss, postoperative pneumonia or atelectasis rate, postop-erative wound recovery, thoracic drainage in postoperative three days, incidence of postoperative atrial fibrillation, perioperative hospital stay were compared between the two groups. Results In operative time, blood loss and thoracic drainage volume in postoperative three days, there was no significant difference in two groups (P>0. 05). In paients with and without DM, the occurrence rate of atrial fibrillation was respectively 50. 00% (3/6) and 16. 66% (1/6), the occurrence rate of postoperative pneumonia or atelectasis was respectively 50. 00%(3/6) and 16. 66% (1/6), and postoperative hospital stay was respectively (16. 28 ± 3. 61) d and (11. 11 ± 2. 92) d. It was of significant differences (P<0. 05). Conclusion Compared with NSCLC patients without DM, postoperative atrial fibrillation and pneumonia or atelec-tasis occured more often in the DM cases. And patients with DM have longer hospital stays.