OBJECTIVE To establish SYBR Green Ⅰ fluorescent quantitative PCR assay for the detection of West Nile virus(WNV),which could be used for early laboratory diagnosis.METHODS A fragment of WNV gene was amplified by PCR,then cloned into pMD-18 T vector.The combinant plasmid was sequenced and analyzed by means of BLAST program,and used as the positive DNA in place of WNV.The SYBR GreenⅠfluorescent quantitative PCR assay was established based on positive plasmid.The sensitivity and specificity of the assay were performed.RESULTS The combinant plasmid was confirmed by sequencing and the fragment belonged to WNV.Ten copies of WNV RNA were detected by SYBR GreenⅠfluorescent quantitative RT-PCR assay.Results of the other members of Flaviviridae were negative,which indicated this assay was specific for WNV.CONCLUSIONS SYBR GreenⅠfluorescent quantitative PCR assay established in this study is highly sensitive and specific,and so it can be used for early diagnosis of WNV infection.

In this paper, intelligent control of ultrasonic power during the process of thrombus ablation is realized through the combination of BP Neural Network and the expert system of ultrasonic thrombus ablation. One proper real-time power can be output correspondingly to different treatment circumstance according to expert ruler during the process of treatment.

Objective To investigate the effect of continuous positive airway pressure therapy (CPAP) on heart rhythm and blood pressure variability in primary hypertensive patients with obstructive sleep apnea/hypopnea syndrome (OSAHS).Methods Ninety seven patients with OSAHS and primary hypertension were randomly divided into treatment group (n =49) and control group (n =48).Patients in control group receive antihypertensive medication only,while patients in treatment group received antihypertensive medication in combination with CPAP therapy for 4 weeks.The dynamic electrocardiogram (ESG) and ambulatory blood pressure for 24 h were measured before and after treatment in both groups.Results After CPAP treatment,the night index of standard deviation of the average NN intervals (SDANNniGHt) in treatment group was (89.0 ± 21.4) ms,while the day and night index of square root of the mean squared differences of successive NN intervals (rMSSDday and rMSSDnight) was (35.9 ± 17.2) ms and (45.7 ± 18.3) ms respectively.Compared with those before treatment,the index of SDANNnight was decreased and the indexes of rMSSDday and rMSSDnight were increased (P ＜ 0.01).Compared with control group,the SDANNnight was decreased,and the rMSSDday and rMSSDnight were increased (P ＜ 0.01,P ＜ 0.05).After CPAP treatment,the night index of low-frequency power (LFnight) in treatment group was (15.3 ± 5.7) ms,which was lower than that before treatment,and the night index of high-frequency power (HFnigh) was (16.3-± 8.1)ms,which was higher than that before treatment (P ＜ 0.01);the LFnight in treatment group was decreased and the HFnight was increased (P ＜ 0.01).After CPAP treatment,the day and night indexes of systolic blood pressure (SBPday and SBPnight) and the night indexes of diastolic blood pressure (DBPnigh) t in treatment group were (136.1 ± 13.4) mmHg,(128.7 ± 9.8) mmHg (1 mmHg =0.133 kPa),and(83.7 ±7.2)mmHg respectivley,which were all lower than those in control group (P ＜ 0.05 or P ＜ 0.01).The incidences of morning peak blood pressure and non-dipper hypertension in control group were significantly higher than those in treatment group (P ＜ 0.01).Conclusion CPAP therapy can effectively adjust the heart rate and blood pressure levels in patients with OSAHS and primary hypertension,especially in the night.

Objective To access the value of the combination of axial image,MPR,and VE in theevaluation of laryngeal carcinoma involvement.Methods Twenty patients with laryngeal carcinoma or hypopharyngeal carcinoma were prospectively studied by helical CT,and MPR and VE were subsequently done on the Voxel Q workstation.The axial images findings and the combined image findings of axial image,MPR,and VE were compared with the pathological results by suing a predetermined checklist of 17 regions according to the TNM classification of malignant tumors(UICC and AJCC).the results were studied in a blind way. Results In the evaluation of the neoplastic invasion of ventricular fold,vocal cord,the anterior commissure,subglottic region,thyroid cartilage,and tissue beyond the larynx,the combined image were better than axial image in sensitivity(100% vs 92.4%,P=0.064),specificity(98.5% vs 89.5%,P=0.028),and accuracy(99.2% vs 90.8%,P=0.003).Neoplastic invasion of the arytenoid cartilage was present in 6 patients.The sensitivity and the specificity was 83.3% and 100% respectively when using the criteria of the arytenoid cartilage sclerosing combined with distortion,erosion or lysis.The specificity was only 57.1% when using the criteria of arytenoid cartilage sclerosing for judging parameter.The result was identical when assessing the arytenoid cartilage,PGS,and PES between the two groups.Conclusion Axial image combined with subsequent MPR and VE could improve the diagnosis in the evaluation of the neoplastic invasion of ventricular fold,vocal cord,the anterior commissure, subglottic region,thyroid cartilage,and tissue beyond the larynx.

ObjectiveTo explore the effects of different nutrition methods on the number and activity of natural killer (NK) cells in liver after hepatectomy in cirrhotic rats. MethodsTwenty thioacetamide-induced cirrhotic rats were randomly divided into enteral nutrition (EN) group ( n =10) and parenteral nutrition (PN) group ( n =10), in which EN or PN was administered for 5 consecutive days 1 day after hepatectomy. The percentage of the NK cells in livers was examined by flow cytometry. Activity of NK cells was measured by 4 h51Cr-release assay.ResultsCompared with the PN group, the percentage (22.2 ％ ± 1.7％ vs. 17.4％ ± 1.8％, P =0.000) and the activity of NK cells (41.6％ ± 2.3％ vs. 37.1％ ± 2.2％, P =0.000) in the livers were significantly higher in the EN group. ConclusionEN after posthepatectomy can ameliorate the percentage and the activity of NK cells in the livers in cirrhotic rats.

Objective To investigate the role of p21-activated kinase 1 (PAK1) in colorectal mucosal carcinogenesis and the relationship between PAK1 expression and biological behavior of colorectal carcinoma. Method PAK1 was detected with immunohistochemical method in 10 normal colorectal mucosas,40 colorectal villous or tubular adenomas and 60 colorectal carcinomas. Results The positive rate for PAK1 was 10.0% in normal colorectal mucesas,and 25.0%, 33.3% and 33.3% in slight, moderate and severe dys-plastic adenomas, respectively, 65.0% was found in colorectul carcinomas. The positive rate for PAK1 in col-orectal carcinomas was higher than that in normal colorectal mucosas(P<0.01), colorectal villous or tubular adenomas(P<0.01), slight dysplastic adenomas(P<0.01) and moderate dysplastic adenomas (P<0.05).The positive rate for PAK1 of poor differentiated colorectal carcinomas was higher than that of high differentiated ones (P<0.05), and the positive rate for PAK1 of patients with lymph node metastasis was higher than that of patients without lymph node metastasis (P<0.05). In clinical stages, the positive rate for PAK1 in Dukes C and D stages patients was higher than that in Dukes A stage patients, respectively (P<0.05). Conclusion PAK1 maybe play some role in the process of carcinogenesis of colorectal mucesa, and be used as an useful marker for assessment of the biological behavior and prognosis of colorectal carcinoma.

STAT-3 and Ki67 protein were significantly higher(P＜0.05)in the EN 5 days group.Conclusion Post-hepatectomy EN in cirrhotic rats can speed up liver function recovery,prevent cholestasis,and promote liver protein synthesis and liver regeneration.

AIM: To compare the difference in serum prolactin levels and the concentrations of anti-infective factors of human colostrums between normal labor and cesarean section. METHODS: The serum prolactin levels were measured at 48 h, 96 h after parturition in 100 cases of normal labor and cesarean using radioimmunoassay. Furthermore, the levels of SIgA, IgG, IgM, C3, C4 were tested in 100 cases at the same time using the technique of immuno-turbidimetry. RESULTS: The serum prolactin levels in normal labor tested at 48 h after parturition were significantly different with that of the cesarean section group. But there was no significant difference in serum prolatin levels 96 h after parturition between the two groups. The levels of SIgA,IgG,IgM,C3 and C4 of the colostrum in the cesarean group were higher significantly than those of the normal group at 48 h after parturition. (P＜0.05 or P＜0.01). At 96 h after parturition, there was no significant difference in SIgA, IgG, IgM and C3 between the two groups except C4. CONCLUSION: At 48 h after paturition, the serum prolactin levels of normal labor group are higher than that of the cesarean section group. The concentrations of anti-infective factors (SIgA, IgG, IgM, C3 and C4) of human colostrums at 48 h after paturition in cesarean section group are higher than those of normal labor group.

Objective To explore the difference of the level of Th1/Th2 balance and change of CD4+ CD45R cell after treated by laparoseopy-assisted rectal cancer surgery and conventional open rectal cancer surgery, so to compare the effect of the two procedures on immunity function of the patients. Methods CD4+ IFN-γ+ T cell (Thl cell), CD4+ IL-4 + T cell( Th2 cell)and the ratio of Th1/Th2 were detected by flow cytometery ; The levels of IFN-γand IL-4 were measured by ELISA. The changes of CD4 + CD45RA+ T cell and CD4+ CD45RO+ T cell were detected in the different procedures so that the effect of the two procedures on eytoimmunity and immune balance were analyzed. Results Compared with baseline levels, the level of Th1 cells was decreased [ (4.51±1.52 ) %, (7.26 ±2.59) % vs. ( 12.06 ±1.82 ) % ] (P < 0.05 ) ; The level of Th2 cells was increased [ (6.70 ± 2.41 ) %, (6.70 ± 2.41 ) % vs. (4.47 ± 1.90) % ] ( P < 0.05 ) ; The level of IFN-γ was decreased [ (57.15± 23.64) ng/L, (72.70 ± 27. 31 ) ng/L vs. ( 123.47 ± 32. 52) ng/L ] ( P < 0. 01 ) ; The level of IL-4 was increased [(55.55±7.29)ng/L,(57.56 ±7.13)ng/L vs. [(41.87±5.83)ng/L] (P<0.05) on 2nd and 7th day after conventional open reeatal cancer surgery; The level of Thl, Th2, IFN-γ and IL-4 in the laparoscopy-assisted rectal cancer surgery was not different ( P > 0.05 ). At 7 day postoperatively, the level of the CD4+ CD45 RA+ T cells was in-creased (35.11 ±7.82) and the CD4+ CD45RO+T cells was decreased(61.21±9.13) compared with surgery be-fore(31.11±6.72) and (68.11 ±11.42) respectively (P< 0.05). Conclusions The laparoscopy-assisted rec-tal cancer surgery has less influence on immune balance, thus protect systemic cell immunity.

OBJECTIVE To detect serotype of dengue virus in sera from patients with fever.METHODS A pair of degenerated primers and one MGB probe were designed targeting the conserved region at the E gene of dengue type 1 virus.TaqMan MGB real-time PCR assay was developed with plasmid including E gene of dengue type 1 virus as standard sample.The sera of 10 patients with fever were used to extract RNA,and convert into cDNA.Then cDNA were detected by TaqMan MGB real-time PCR assay and the amplified products were analyzed at the same time.RESULTS The sera of 9 patients from 10 samples were observed to generate a fluorescent signal,and about 100 bp fragment was obtained simultaneously.CONCLUSIONS Dengue fever on 2006 in Guangzhou is caused by the dengue type 1 virus.TaqMan MGB real-time PCR assay is rapid and sensitive to detect dengue virus infections.