Résumé
Objective:To prepare rabbit polyclonal antibodies against respiratory syncytial virus (RSV) N protein and use them as the detection antibodies to establish a N-ELISA-based method for rapid detection of neutralizing antibodies.Methods:A plasmid of pET30a-N for the expression of RSV N protein was constructed. After purification, the protein was immunized into New Zealand rabbits to prepare polyclonal antibodies, which were used as the detection antibodies. Positive serum samples were diluted and used to neutralize RSV (100 TCID 50/well). Hep-2 cells were inoculated and cultured, and then the cells were fixed with 80% acetone. ELISA was performed to detect RSV N protein in infected cells. When the absorbance value of a well was below the cut-off value, it was regarded as the positive well in the neutralization test. The highest dilution of a positive well serum was the neutralizing antibody titer. After optimizting the antibody dilution, detection time, cell density and the duration of neutralization, the method for neutralizing antibody detection was established based on N-ELISA. The established method was verified by analyzing the influences of different cell generations and edge effects, and calculating the accuracy, repeatability and precision. The correlation between the established method and microneutralization method was analyzed by detecting human RSV IgG-positive serum. Results:The plasmid pET30a-N was successfully constructed, and the expressed N protein showed high purity and good specificity. After the third immunization, the antibody titer in rabbit serum was 1∶51 200, and the antibodies could specifically bind to RSV. The prepared rabbit anti-RSV N polyclonal antibodies had a titer of 1∶51 200, and showed good specificity. The neutralizing antibodies could be detected on day 4 with the established method, and the duration of neutralization was shortened to 30 min. Cell generations and the position of wells in the 96-well plate (edge well and non-edge well) had no significant effect on the method, and the repeatability, precision and accuracy of the method were good. In the detection of 64 RSV IgG-positive human serum samples by the established method and microneutralization method, the correlation coefficient was 0.929 6, indicating a good positive correlation between the two methods.Conclusions:A N-ELISA-based method for rapid neutralizing antibody detection is successfully established, which can be used to evaluate the serum antibody level after RSV vaccination.
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Abdominoscrotal hydrocele is rare in clinic. In the past, routine examination and diagnosis were difficult, easy to be misdiagnosed. The daily operations were mostly completed through the groin area or abdominal incision, the wound is large. The application of laparoscopy can clearly diagnose the abdominoscrotal hydrocele through "springing back ball" sign, and can cure the disease by laparoscopic resection of interperitoneal mass and closure of the internal ring. It is worthy of clinical application. In this article, we summarized and analyzed the clinical experience of 15 cases of children with abdominoscrotal hydrocele diagnosed and treated by laparoscopy, to explore the value of the laparoscopic technology in the diagnosis and treatment of the abdominoscrotal hydrocele.
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Objective To compare the clinical results of the ureteral reimplantation with the traditional open approach and laparcocopic pneumovesical approach. Methods A retrospective review of 108 patients who underwent ureteral reimplantation from December 2004 to October 2010 was conducted.The patients were divided into open and pneumovesical groups according to the surgical approach.Perioperative results were compared between the two groups in terms of operative time,postoperative intravenous antibiotics duration,catheterization duration,postoperative stay,average total cost and surgical cost,respectively.And the pneumovesical group was divided into two stages by time,compared the operative time of the two stages and between the later stage and the open group. Results The postoperative hospital stay of pneumovesical group was shorter than the open group (6.8 ± 1.9 d and 8.9 ± 2.9 d,P =0.002 ).For catheterization duration,the pneumovesical group was shorter than the open group as well (5.2 ± 1.2 d and 6.2 ±2.2 d,P=0.057).For the postoperative intravenous antibiotics duration,the pneumovesical group was 5.0 ± 1.3 h,the open group was 5.4 ±1.6 h (P =0.159).For the total cost,the pneumovesical group was 16 067.9 ±4 295.8 RMB,the open group was 15 617.7 ± 5 486.5 R MB (P =0.168).For the surgical cost,the pneumovesical group was 9369.4 ± 1366.6 RMB,the open group was 7397.9 ± 1797.3 RMB ( P =0.083 ).Operative duration of the pneumovesical group and open group were 3.2 ± 1.1 h and 2.3 ± 1.1 h ( P =0.003).For pneumovesical group,the mean operative durations of the two stages were 3.6 h and 2.8 h (P =0.286).And the later stage of pneumovesical group was a little longer than the open group,but no significant difference ( P =0.234 ).No major complication was found in the 2 groups during the operative time and the postoperative hospital stay.Twenty-four patients (38 ureters) of the pneumovesical group were followed up with micturating cystourethrography ( MCU),ureterovesical reflux recurred in 3 patients.Two patients changed from grade Ⅲ to grade Ⅰ and 1 patient changed from grade Ⅴ to grade Ⅲ after the surgery.Five patients (9 ureters) of the open group were followed up,1 patient found bladder diverticulum; 1 patient found ureteral stricture 6 months after the surgery and got improved after secondary ureteral relimplantation surgery. Conclusions The pneumovesical approach is shorter than the open group in postoperative hospital stay and catheterization reserved duration.The pneumovesical approach is a safe and effective option for ureteral reimplantation.
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In this study, 38 experimental subjects of diabetes mellitus were given pectin 25g extra per day orally under carefully controlled dietary plan for six weeks. Another 38 cases were selected at random as control group.Pre- and posttreatrnent, blood and urine sugar and serum insulin of each subject were measured and compared. The results were: 1) the blood sugar dropped markedly as compared with that of pretreatment (p05). 2) sugar in urine decreased evidently in contrast with control group, but no difference was seen in the volume of urine between the two groups. 3) serum insulin levels dropped significantly when compared with control group (p