Objective To study the effect of small interfering RNA(siRNA) of human papillomavirus(HPV) 18(E6) gene on apoptosis of HPV-related cervical HeLa cell line.Methods siRNA targeting HPV18 E6 mRNA was designed and generated by PCR amplification.The PCR products containing U6 promoter and the siRNA sequence were then transfected into HeLa cells via Lipofectamine()~(TM)2000.Cell viability was determined by MTT assay.Apoptosis was detected by morphological observation and flow cytometry analysis.The expression level of HPV18 E6 mRNA was assayed by RT-PCR.Results The cell growth and viability of(siRNA) transfected group were significantly inhibited(P

Objective To observe the time-intensity curve characteristics of contrast agents in intraocular tumor.Methods A total of 236 patients (238 eyes) with intraocular tumor were enrolled in this study.All the patients received regular ophthalmologic examination,two dimensional ultrasound,color doppler ultrasonography and contrast-enhanced ultrasonography.There were 166 patients (166 eyes) with choroidal melanoma,16 patients (18 eyes) with choroidal metastatic carcinoma,52 patients (52 eyes) with choroidal hemangioma,two patients (two eyes) with retinal hemangioma.The whole process of contrast-enhanced ultrasound were recorded, and exported as t images of Dicom format.These images were processed by Sonoliver software (Tomteck Company, Germany) to drawn the time-intensity curve of contrast agents in the intraocular tumors.Results All intraocular lesions were completely filled with contrast agent,concentric filling from the periphery to the center can be documented in some cases.The time-intensity curve of choroidal hemangioma and retinal hemangioma were basically the same.The time-intensity curve of choroidal melanoma and choroidal metastatic carcinoma were also basically the same.In the filling phase,all tumors were rapid filling type.In the regression phase,contrast agent subsided earlier than in control tissue within the melanoma or metastatic carcinoma lesions,but subsided synchronous or slightly faster than in control tissue within the choroidal hemangioma and retinal hemangioma lesions.Among 166 eyes with choroidal melanoma,138 eyes (83.1％) were in full compliance with the above changes,28 eyes (16.9％) were largely in line with these changes.All the eyes (100.0％) with choroidal metastatic carcinoma,choroidal hemangioma and retinal hemangioma were in full compliance with the above changes.Conclusion Time-intensity curve is quickly filling and fast regression for malignant intraocular tumors,but is quickly filling and slow regression for benign intraocular tumors.

Objective To study the double restriction fluorescent labeling (DRFL) method for fluorescent labeling of trace DNA samples and its effect in enhancing the pathogen detection sensitivity of microarray assays. Method SARS-CoV RNA samples were reversely transcribed and then further amplified with the restriction display (RD)-PCR and fluorescently labeled by conventional restriction labeling directly with Cy-universal primer and the novel double labeling with Cy-universal primer and CydNTP. The labeled samples were applied to the microarray with the viral probes, processed and analyzed. Results Compared with the conventional method, DRFL labeling resulted in 3. 5835 times higher fluorescent intensity of all the SARS probes on average, even though increased fluorescent intensities for different probes varied considerably. Conclusion Signal to noise ratio can be enhanced by the DRFL method which improves the sensitivity of microarray technology in trace pathogen detections.

Objective To determine the relation between enlarged perihepatic lymph node (PLN) and viraemia,and to find out whether there is a difference in PLN size between the healthy individuals and patients with hepatitis C virus (HCV) infection. Methods Seventy-four patients with HCV infection and 283 healthy individuals were examined by two-dimensional ultrasound. The length and thickness of lymph node were measured. The lymph-node area index(LN area index) was calculated. The LN area index was evaluated by the ROC curve and compared with serum HCV core antigen (HCV-Ag) levels by correlation analysis. Results Detection rate, the length and thickness, LN area index of group with HCV infection were significantly different by compared with the normal control group ( P < 0. 001 ). The ROC curve showed that the LN area index as a diagnostic indicator had high sensitivity and specificity. The LN area index showed a significant correlation with HCV-Ag level (r = 0.436, P <0.05), but no significant differences were found between LN area index and HCV-Ag grade. Conclusions Perihepatic lymphadenopathy indicates viraemia,and LN area index of 100 mm2 is a reference index in estimating whether patients have HCV infection.

Objective: To evaluate the initial pain and influential factors in orthodontic treatment by VAS(visual analogue scales),and explore the way to alleviate the pain. Methods:107 subjects(47 male and 60 female) were selected. Following insertion of the first archwire (0.36 mm Chinese NiTi wire), visual analogue scale(VAS) was used to record the severity of pain at 4,12,24 h and on day 3,5,7. All the subjects were randomly divided into two groups by ligature with elastomeric rings (in 51 patients) or stainless steel wires (in 56 patients).In all the 107 patients, 30 people need to extract their tooth for relieving crowding while the other 77 were non-extraction cases. Results:No significant differences of pain were found between elastomeric rings ligature cases and stainless steel wires ligature cases at each stage; as well as between extraction and non-extraction cases. But the adults perceived more pain than the adolescents. Patients who were asked to chew gum or to do something (watching TV or listening to music) for distraction perceived less pain.The differences were statistically significant between different groups. Conclusion:The orthodontist should make a good communication with patient about initial pain before the orthodontic treatment, especially for adults.

Objective To discuss the diagnosis characteristics and clinical value of posterior vitreous detachment (PVD) by color Doppler flow imaging(CDFI). Methods One thousand and two cases were examed by CDFI for the diagnosis standard and property of PVD. Results Typical acoustic properties of PVD: the B scan may show a very fluid thick line. There were two types complete PVD and part PVD based on the vitreous separated from the optic nerve. Kinetic echography typically showed a very fluid, undulating after movement of the completely PVD. Part PVD had more than one dot with attachments to the retina and less mobile than complete PVD. There was not blood flow signal on the PVD by CDFI. Conclusions Ultrasonography for PVD not only exactitudes,but also provides foundation for clinic.

Endothelium-derived hyperpolarizing factor (EDHF) is the third factor released by en-dothelial cell other than NO and PGI2. It relaxes smooth muscle accompanied by a hyperpolarization in the membrane potential. EDHF may be epoxye-icosatrienoic acids (EETs) formed from arachidon-ic acid by the action of cytochrome P450. It is synthesized and/or released by endothelial cell as a result of an cytosolic Ca2+ increase, which is stimulated by the action of acetylcholine or bradykinin on endothelial cell. EDHF is shown to activate Ca2+-activated K+ channels and induce a hyperpolarization in the membrane potential in vascular smooth muscle. The hyperpolarization of the membrane inhibits the opening of voltage-dependentcalcium channels, allows calcium sequestration and removal mechanisms to lower intracellular calcium, and leads smooth muscle to relaxation. In large conducing arteries, EDHF may provide a secondary system to NO, which assumes primary importance in endothelium-dependent relaxation and inhibits the release of EDHF. However, in small resistance arteries, EDHF appears to be a major determinant of vascular calibre, and may be of primary importance in the regulation of vascular resistance.

Objective To investigate the therapeutic effect of intra-arterial thrombolysis (IATT) in acute cerebral infarction(ACI). Methods 15 patients with ACI were treated with IATT using Urokinase(UK). All patients were assessed by DSA before and after IATT. The stroke scale(ESS and NIHSS) and Barthel Index(BI) were used before and 2 h,48 h,15 d ,30 d after treatment for appraising thearpeutic effect . Results After IATT, the sides of obliterated middle cerebral artery in 12 patients were completely re-circulated. Compared with before IATT, the scores of ESS, NIHSS and BI were significantly improvement at 2 h,48 h,15 d and 30 d after IATT(P

0.05). There was no correlation between Bcl-2 and Fas positive cells and acetylcholine receptor antibody serum titre, disease grouping, age and sex in patients with or without hyperplastic thymic and thymoma.Conclusions In MG patients, the expression of Bcl-2 in hyperplastic thymus is upregulated. There is no correlation between the expression of apoptotic genes Bcl-2 and Fas and clinic relative factors.

Objective To design a microarray of ~60mer oligonucleotide for detection and sub-typing of human papillomavirus (HPV). Methods The type-specific oligonucleotide probes of 4 different types of HPV (6, 11, 16, 18) were designed by using biological software Arraydesigner 2.0, which analyzed the whole genome sequences of HPV and selected optimal probes with high specificity, identical length and similar melting temperature (Tm). These probes were synthesized and printed onto the surface of glass slides in order to prepare a low-density microarray. HPV samples were labeled with fluorescence dyes Cy3 using a method of restriction display PCR (RD-PCR). HPV plasmid DNA was restricted with Sau3AⅠ to produce multiple fragments which were ligated to adaptors subsequently and used as PCR template. PCR labeling was performed with the fluorescently labeled universal primer (Cy3-UP) whose sequence is designed according to the adaptor of the RD-PCR approaches. The labeled samples were then hybridized with the oligonucleotide microarray. Results Both single and multiple HPV DNA samples could be detected with oligonucleotides microarray, and the corresponding HPV subtypes were recognized as well. And no signals were detected in all the negative and blank control spots. Conclusion 60mer oligonucleotide microarray designed by appropriate bioinformatics software can be applied to HPV detection and genotyping on gene level.