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.

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.

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.

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.

Objective To investigate the blood fatty levels in different genotypes of matrix metalloproteinases (MMP) and the carotid atherosclerosis (CAS) in patients with essential hypertension (EH).Methods Four hundred and eighty-eight cases of Han and Uygur in patients with EH were selected,and MMP-2-735 C/T and MMP-3-1171 5A/6A polymorphism was measured by polymerase chain reactionrestriction fragment length polymorphism.They were divided into two groups:EH with CAS group (CAS group,293 cases,Han 171 cases,Uygur 122 cases) and EH without CAS group (NS group,195 cases,Han 105 cases,Uygur 90 cases).Blood fatty factors were analyzed according to the different MMP genotypes.Results According to MMP-2 genotypes,in Hart CAS group,the low density lipoprotein cholesterol (LDL-C) levels in MMP-2 CT+TT genotypes were significantly higher than those in CC genotype(P ＜ 0.05),and high density lipoprotein cholesterol levels were significantly lower than those in CC genotype(P ＜ 0.05); In Uygur CAS group,the triglyceride levels in MMP-2 CT+TT genotypes were significantly higher than those in CC genotype(P ＜ 0.05).According to MMP-3 genotypes in Han NS group,the total cholesterol and LDL-C levels in 6A/6A genotype were significantly higher than those in 5A/5A+5A/6A genotypes (P ＜ 0.05).Conclusion The blood fatty levels are different in patients with different MMP genotypes,which may probably influence the CAS.

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 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.

Objective To evaluate specific clinical and imaging features of osteogenesis imperfecta V and to improve diagnostic accuracy of this disease. Methods Data of 15 patients with osteogenesis imperfecta type V were retrospectively analyzed for their clinical and imaging features. There were 10 males and 5 females, aged from 1 year and 30 years old (median age,12.5 years ). All 15 patients had plain X-ray, and 4 of 15 had CT. All data were analyzed by 3 experienced deputy chief doctors in OI according to OI V standard. Results X-ray:calcification of the interosseous membrane between radius-ulna was detected in all patients and calcification of the interosseous membrane between tibia-fibula was detected in 2 of 15 patients. Dislocation of the radial head was seen in 13 of 15 patients,bilateral in 9 and utilateral in 4.All patients showed restriction in the pronation and supination of the forearm and restricton in the flexion and extention of the elbow joint. Patients with dislocation of raidal head were associated with large coronoid process and olecranon of the ulna. Hyperplastic callus of the extremities were detected in 7 of 15 patients (7 at femur , 3 at humerus, 1 at tibia.In early stage, hyperplastic callus showed thin cortice, and clear boundares with the diaphysis showing and low density, irregular, mesh-like lamellation inside. In the later stage, there were dense calcification inside hyperplastic callus, and no difference in density with the diaphysis. Diaphysis surrounded by hyperplastic callus had clear boundaries with the hyperplastic callus. No cortical destruction was detected. CT:there were sparse needle-dot calcification inside hyperplastic callus, with the patterns of granular, ring-and-arch,irregular streaky mineralization. The cross section of proximal femoral shaft showed irregular shape , flat square shape and tiny medullary cavity, with no calcification on the edge of hyperplastic callus. CT value:-91 HU inside hyperplastic callus; 283 HU in femoral shaft. Conclusions Interosseous membrane between radius-ulna or tibia-fibula, hyperplastic callus ,dislocation of the radial head are specific features in osteogenesis imperfecta V. X-ray can make a definitive diagnosis of osteogenesis imperfecta V. CT scan is helpful in the differential diagnosis of osteogenesis imperfecta V from osteosarcoma.

Objective To investigate the impact of a single sub-anesthetic dose of ketamine on postoperative emotional reactions of anxiety and depression during colorectal tumors surgery. Methods A total of 42 patients undergoing selective colorectal surgery,aged 18 to 65 years,were randomly divided into ketamine group (group K)and control group (group C).After induction of an-esthesia,a single intravenous injection of 0.3 mg/kg ketamine and saline were used in Group K and group C 5 minutes before the operation respectively.The intravenous analgesia program was identical between the two groups.The patients??emotional reactions (anxiety,depression)were assessed using the Hospital Anxiety and Depression Scale (HAD),the quality of recovery was assessed using the QoR-40 questionnaire on the day before operation and within postoperative 48 hours respectively.Pain was estimated by the visual analog scale (VAS)and sedation was assessed with Ramsay score 30 mi-nutes after extubation.The time of anesthetic end and extubation were recorded.The complications during anesthesia and recovery such as cough, agitation 30 minutes extubation were recorded. Results The HAD-A and HAD-D scores of group K were lower than group C (P <0.05)48 h post-operatively.There was no difference on the QoR-40 score postoperative 48 h between the two groups. The patients??emotional state QoR-40 score of group K were higher than group C (P <0.05 ).The VAS scores 30 minutes after extubation of group K were lower than group C (P <0.05).There was no significant difference on sedation score 30 minutes postoperatively between the two groups.There was no significant difference in extubation time,cough,agitation and delirium between the two groups.There was no dizziness, nausea, vomiting or diplopia 30 minutes after extubation. Conclusion A single subanesthetic dose of ketamine can significantly reduce the scores of postopera-tive anxiety and depression,improve the quality of recovery,and no postoperative adverse reactions increased.