Objective To establish a specific,sensitive and applied method for the detection and differentiation of dengue virus types Ⅰ-Ⅳ,Japanese encephalitis virus and yellow fever virus.Methods Based on the genomes sequence analysis,6 pairs of primers were designed.The special capture probes of dengue virus types Ⅰ-Ⅳ,Japanese encephalitis virus and yellow fever virus were amplified,cloned and sequenced.Then the microwell plates were precoated using these capture probes,and the forward primers were labeled using biotin.The samples were then amplified using the biotin labeled forward primers and reward primers.The microwell plate hybridization was processed for detecting and differentiating the virus.The precoated DNA concentration,precoated time,hybridization temperature and hybridization time were optimized carefully.Results The A value of positive samples were over 0.5,while the average A value of the negative samples was less than 0.1.The S/N value exceeded 10.0.Sensitivity experiment suggested the method of PCR-ELISA could detect the virus RNA in 107 times dilution,while RT-PCR could detect the virus RNA in only 106 times dilution.The stability experiment of PCR-ELISA using DVⅠ suggested that the within-batch coefficient of variation was 6.21%,the between-batch coefficient of variation was 9.92%;the within-batch coefficient of variation in negative control was 1.92%,and the between-batch coefficient of variation in negative control was 3.68%.No visible changes were found on the performance of the coated microwell plates when stored in 4℃for 6 monthes.Conclusion PCR-ELISA is a more sensitive and specific method than RT-PCR is in the early detection and type identification of dengue Ⅰ-Ⅳ types virus,Japanese encephalitis virus and yellow fever virus.

Twenty five patients with incarcerated stone in distal common bile duct were treated with mini-blasting lithotrite under choledochoscope from June 2008 to September 2010.The clinical data of patients were retrospectively analyzed.The successful rate of surgery was 100％.The clearance rate of first procedure was 92％ (23/25),2 cases underwent second procedure and all were successful.There were no severe operative complications including bile duct injury,perforation,hemorrhage or bile leakage.Miniblasting lithotrite is a safe and effective method for treating incarcerated stone in distal common bile duct.

A questionnaire was used to survey the perspective of the 22 mentors,24 students, 27 graduate students and 27 public health units on the competence of MPH student. The result showed that the practice skill,analyzing skill,program plan and practice skill,and emergency reaction abil-ity got more attention. This suggested that to the training of MPH,the training objective should focus on the ability to resolve problems; the curriculum should focus on the characters of public health and on the practice training.

OBJECTIVE:To provide reference for the implementation of Quality Management Guidance of TCM Trust-agent Decocting in Medical Institutions in Shanghai. METHODS:Stratified random sampling method was adopted for the questionnaire of TCM trust-agent decocting in 13 medical and health institutions in the suburb of Shanghai. RESULTS:A total of 13 question-naires were sent out and 13 valid questionnaires were received with the effective rate of 100%. In the 13 medical and health institu-tions,12 medical and health institutions partially or fully adopted trust-agent decocting,and the trust-agent decocting in the 12 med-ical and health institutions accounted for 92.3% in all trust-agent decocting;the 12 medical and health institutions with TCM trust-agent decocting service for patients included 9 pharmaceutical factories and 3 pharmacies;label related problems (including name error,oral and topical label error and stickers unclear) and leakage were the main quality problems,accounting for 33.0%and 31.0%,respectively;83.3% of the medical and health institutions didn’t establish the file of the quality control for TCM trust-agent decocting;decocting process was the most important factor that affects the quality. CONCLUSIONS:Implementing a re-gional Quality Management Guidance of TCM Trust-agent Decocting in Medical Institutions in Shanghai and establishing a system that drug administration,industry associations and medical and health institutions are participated in have great significance to pro-mote the healthy development of TCM trust-agent decocting service and ensure patients’medication safety.

ObjectiveTo clarify the clinicopathological features of renal amyloidosis in order to achieve early diagnosis and treatment.MethodsClinicopathological data of 26 biopsyproven renal amyloidosis cases in Department of Nephrology,Zhongshan Hospital,Fudan University between2006and2010wereanalyzedretrospectively.Immunohistochemistryand immunofluorescence of amyloid A protein,immunoglobulin light chains such as K、λ were performed on renal specimens for further classification.ResultsAge of 26 patients ranged from 40 to 77 years old,average(58.54±10.07) years.Twenty-two out of 26 patients(84.62％) were treated in local hospital before admitted to our department,and 21 patients(95.45％) were misdiagnosed as chronic primary glomerulonephritis.The prominent clinical manifestations of renal amyloidosis were nephrotic syndrome(17 cases,65.38％),decreased blood pressure(16 cases,61.53％),organ enlargement (8 cases,30.77％) and bodyweight loss (6 cases,23.08％).Fourteen out of 25 patients (56.00％) were found to have monoclonal light chains in serum by immunofixation electrophoresis.Three patients with mild pathological changes who had no confirmable Congo red stain were conffimed by electron microscopy. Twenty-three(88.46％) patients werediagnosed as AL amyloidosis,one(3.85％) as AA amyloidosis,one was strongly suspected of hereditary amyloidosis,and one was undetermined.ConclusionsRenal amyloidosis is frequently misdiagnosed.Middleaged and old nephrotic patients with decreased blood presure,organ enlargement and bodyweight loss may be the most helpful clues of the disease.Most patients have monoclonal light chains in serum or urine.Renal biopsy,especially electronic microscopy plays a crucial role in the early diagnosis of renal amyloidosis.Immunohistochemistry is important for patients with renal amyloidosis in pathological classification and treatment.

Objective:To study the effect of OSAHS on adult nasal-cycle.Method:The nasal cycle of 20 healthy adults,18 patients of snoring and 22 patients of OSAHS were examined with acoustic rhinometry,which was performed every 30 minutes in 7 hours.Result:The ratio of nasal cycle in healthy adults was(19/20,95.0%), in snoring patients was(15/18,83.0%),in patients of OSAHS was(15/22,68.2%). The mean alteration amplitude of nasal cycle in healthy adults was significantly larger than that in patients with OSAHS (P<0.05).The distribution of the healthy adults and patients with OSAHS between the typical cycle categories was significantly different(P<0.05).Conclusion:The characteristics of nasal-cycle in of OSAHS patients were different with healthy adults,which maybe owing to change of physical function of nasal cavity.

Objective To compare the diagnostic efficiency between blastomere preimplantation genetic diagnosis (PGD) and polar body PGD for chromosomal translocation carriers. Methods Group A had 8 cycles using whole painting probes for the first polar body diagnosis, while group B had 29 cycles using two subtelomeric probes and one centromeric probe for the blastomere diagnosis. Results The fertilization rate of group A was significantly lower than group B [66. 1% (72/109) vs 85.2% (304/357) , P < 0.05]. There was no significant difference in the successful biopsy rate between two groups. However, group A had a significantly higher loss rate during fixation and higher no signal rate after fluorescence in situ hybridization [ FISH, 9. 6% (12/104) vs 1.6% (4/252), 11.2% (10/89) vs 3.0% (7/233) ]. Totally, the diagnostic efficiency in group A (72. 5% ,79/109 ) was significantly lower than that in group B( 89. 8%, 230/256, P < 0. 05 ). Although both the clinical pregnancy rate( 3/7 ) and implantation rate( 22. 2% ,4/18 ) of group A were higher, the differences were not statistically significant ( P > 0.05 ). Conclusion Both methods can be used efficiently in the PGD for chromosomal translocation carriers. Blastomere PGD has a higher diagnostic rate.

Objective To study the role of hydrogen sulfide (H2S) in the effect of SB203580 on proliferation and apoptosis of hepatic stellate cells and the effects of H2S on expressions of collagenⅠand collagenⅢmRNA in hepatic stel-late cells. Methods There were five groups of HSC-T6 cells in this study including control group (DMEM medium contain-ing10%fetal bovine serum), dimethyl sulfoxide (DMSO) group, sodium hydrosulfide (NaHS)group,SB203580 (SB)group and SB+NaHS group. MTT method was used to detect the cell proliferation and inhibition rate of HSC-T6 cells treated by SB203580 and H2S. The apoptotic rate of HSC-T6 cells was detected by flow cytometry with annexin V-FITC/PI double staining. RT-PCR was used to detect the expressions of collagenⅠand collagenⅢmRNA in HSC-T6. Results The apop-totic rate of HSC-T6 cells was significantly higher in SB group and SB+NaHS group than that of control group, and the rate was significantly higher in SB+NaHS group than that of SB group (P＜0.05). There was no significant difference in the apop-totic rate of HSC-T6 cells between DMSO and NaHS groups than that of control group. The expressions of collagenⅠand col-lagenⅢmRNA were found in five groups of cells. There was a higher expression of collagenⅠand collagenⅢmRNA in NaHS group than that of control group (P＜0.05). The expressions of collagenⅠand collagenⅢmRNA were significantly lower in SB group and SB+NaHS group than those of control group and NaHS group (P＜0.05). Conclusion H2S activated P38MAPK signal pathway. And P38MAPK was specifically blocked by SB203580 in HSC-T6 cells, which inhibited the cell proliferation stimulated by H2S and promoted the apoptosis.

Objective To investigate the pregnancy outcomes of couples with either maternal or paternal balanced translocations.Methods One hundred and ninety-four couples were divided into three groups based on the kind of translocations:135 with reciprocal translocation,52 with nonhomologous Robertsonian translocations,and 7 with homologous Robertsonian translocations.Past reproductive histories were surveyed.For those who wanted to have their own babies by natural conceptions after knowing their karyotypes as well as the risks of abnormal offsprings,subsequent pregnancy outcomes were recorded.Total pregnancy outcomes were compared between three groups.Results(1)503 previous and subsequent pregnancies were recorded in detail.The pregnancy outcomes are as follows:spontaneous abortions 81.7% (411/503);induced terminations because of fetal abnormalities 3.2%(16/503);birth defects 7.2% (36/503);normal/balanced offsprings 8.0%(40/503).In reciprocal translocations,nonhomologous Robertsonian translocations and homologous Robertsonian translocations,the birth defects rates were 5.7% (20/350),10.9%(14/128)and 8.0%(2/25),respectively(P

Objective To observe the outcomes of one-stage posterior short-level pedicle screw fixation combined with anterior fixation of severe thoracolumbar fractures.Methods A retrospective case series study was performed on 21 patients with severe thoracolumbar fractures stabilized by posterior short-level pedicle fixation combined with anterior internal fixation at one stage from January 2012 to December 2014.There were 16 males and 5 females,at age of 17 and 64 years [(38.7 ± 11.4) years].The involved segments included T11 in 2 patients,T12 in 5,Lt in 6 and L2 in 8.For AO fracture classification,type A fractures were seen in 4 patients,type B in 7 and type C in 10.Thoracolumbar injury classification and severity score (TLICS) was (8.12 ± 0.87) points (range,7-10 points).Frankel neurological performance scale was Grade B in 8 patients,Grade C in 11 and Grade D in 2.Operation time,blood loss,nerve function,kyphosis correction and complications were reported.Results Operation time was (234.5 ±57.3)min (range,180-360 min),and blood loss was (387.4 ± 124.4) ml (range,260-950 ml).Time of follow-up was (19.8 ± 3.5)months (range,14-25 months).Nerve function of 18 patients was improved by at least one Frankel scale.Cobb angle was (4.1 ±5.3)° at postoperative 3 days and (4.0 ± 4.9)°at the final follow-up,showing significant differences from that before operation [(-9.3 ± 4.2) °] (P ＜ 0.05).While the difference of Cobb angle did not differ significantly at postoperative 3 days and at final follow-up.No cerebrospinal fluid leakage,vascular injury,incision infection or nerve function deterioration occurred.Conclusion One-stage posterior short-level pediele screw fixation combined with anterior decompression and bone graft fixation is characterized by short operation time,few blood loss,good correction of traumatic kyphosis and good neurological recovery,indicating a good surgical choice for severe thoracolumbar fractures.