OBJECTIVEThe purpose of this study was to develop RT- PCR assay for Rapidly detect and distinguish between Norovirus genogroup I and genogroup II with a pair of primers.

METHODSA pairs of primers specific to capsid prote in ORF2 gene of G I and G II Norovirus were dsigned according to the published complete genome sequence, with which the RNA of Norovirus was extracted and RT-PCR amplification. The sensitivity, specificity of the RT- PCR assay was estimated and apply it to the detection of Norovirus in clinical specimens.

RESULTSThe results showed that the assay possessed high specificity for Norovirus detection and without any evident cross-reaction with other viruses, including rotavirus, enteric adenovirus and hepatitis A virus. The detection limit of RT-PCR assay for Norovirus G I and G II were up to 100 pg/ml and 10 pg/ml respectively.

CONCLUSIONThe RT- PCR assay provide rapid and sensitive detection of Norovirus G I and G II and should prove to be useful for Norovirus diagnosis in the outbreaks of acute gastroenteritis.

Caliciviridae Infections ; diagnosis ; virology ; DNA Primers ; genetics ; Gastroenteritis ; diagnosis ; virology ; Humans ; Norovirus ; classification ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; instrumentation ; methods

Objective To summarize the experience in the diagnosis and treatment of mucinous cystic neoplasms (MCN) of the pancreas. Methods The clinical data of 20 cases who were diagnosed as MCN of the pancreas in the affiliated hospital of Qingdao university from January, 2003 to June, 2008 were collected, data including clinical manifestations, pathological features, treatment and survival were analyzed retrospectively. Results The clinical manifestations mainly included abdominal pain or discomfort, nausea, vomiting; 11 patients had abdominal tenderness, 6 patients had palpable abdominal mass. All the patients underwent ultrasound and CT scan examinations, 13 patients were diagnosed as having benign MCN of the pancreas, 4 were serous cystadenoma, 3 were mucinous cystadenocarcinoma. The tumors were located in the body and tail of the pancreas, with a mean diameter of 4 - 14 cm. All the 20 cases received surgical treatment. The procedures mainly included pancreaticoduodenectomy and resection of pancreatic body and tail. Pathological examinations confirmed there were 10 patients with benign MCN of the pancreas, 3 were borderline mucinous cystadenomas and 7 mucinous cystadenocarcinomas. After a mean follow-up of 26 months, patients with benign MCN of the pancreas or borderline mucinous cystadenomas were still alive without recurrence, the three year survival of patients with mucinous cystadenocarcinomas was 50%. Conclusions MCN of the pancreas mainly occurred in female, and there was no specific clinical features, preoperative ultrasound and CT scan examinations could help to diagnose this disease. Surgical resection was the only effective method to cure MCN with good prognosis.

Objective To identify the species of sarcosaphagous flies by amplifying cytochrome oxidase subunitI (CO I) gene fragment,combined with morphological characteristics.Methods The DNA of sarcosaphagous flies was extacted by modified Tris balanced phenol-Tris saturated phenol protocol,the amplificationand sequencing of CO Ⅰ fragment were conducted,and the results were then compared with the database for analysis.Results The modified DNA extration method by Tris balanced phenol-Tris saturated phenol could obtain effective DNA of sarcosaphagous flies,and be applied for CO Ⅰ fragment amplification,and thus for identification of the species of sarcosaphagous flies.Conclusion The DNA extracted from sarcosaphagous flies by modified Tris balanced phenol-Tris saturated phenol protocol could be used astemplate for the amplification of CO Ⅰ fragment,and the system could identify the species of sareosaphagous flies after sequencing and alignment with the sequences of database.Compared with traditional identification methods of using morphological characteristics,the current system is more accurate and could be more widely applied.

Objective To construct an electrochemical immunosensor for ultrasensitive determination of myeloperoxidase (MPO) .Methods The electrochemical immunosensor for M PO was prepared by modifying the electrode using Au-graphitized me-soporous carbon nanoparticles(AuNPs@ GMCs) hybrid and immobilizing MPO antibodies onto the glass carbon electrode surface . The effect of experimental parameters on the immunosensor and results comparison with ELISA were investigated .Results The immunosensor was sensitive to M PO with a linear relationship between 2 .000 and 300 .000 ng/mL and a correlation coefficient of 0 .999 ;the detection limit was 0 .5 ng/mL .The correlation coefficient of two methods was 0 .983 .Conclusion The immunosensor can be used for ultrasensitive detection of MPO .

Adolescent ; Bacteria ; isolation & purification ; Child ; Child, Preschool ; Humans ; Infant ; Infant, Newborn ; Respiratory Tract Infections ; microbiology

Objective To study the clinical value of color Doppler ultrasonography in typing tuberculous epididymitis.Methods The appearances of color Doppler ultrasound and the findings on operation were analysed in 33 patients with tuberculous epididymitis.Results Of the 33 patients,epididymis appeared as diffusely and heterogeneously enlarged lesions with increased flow in 2 cases,appeared as nodular lesions in 13 cases including nodi with echofree space in 3 cases, nodi with high-level echo patches in 3 cases, and low echo-level nodi in 7 cases. Multiple lesions in scrotum were detected in 17 cases, of whom epididymis up to 11 cases appeared as diffusely enlarged heterogeneous lesions with flow increased.The sonographic appearancs of tuberculous epididymitis could be divided into 3 types:diffusion type, nodus type and complicated type. Nodus type included 3 subtypes: purulence type, calcification type, and cheese type.The accuracy rate of ultrsound diagnosis was 87.9%.Conclusions Testis is easy to be involved when epididymitis appears as diffusion type, so surgical treatments should be early.Purulence type and complicated type need surgical treatments while calcification type does not. Antituberculous drug treatments can be tried before surgical treatments in cheese type.Sonography of urinary system is helpful for the diagnosis of asymptomatic tuberculosis in urinary system when tuberculous epididymitis is first suspected on sonography.

GLuc (Gaussia luciferase) is a secreted luciferase with high sensitivity. In this study, we primarily compared expression character of PTTR with that of PCMV, relied on easy secretion, high sensitivity and simple and fast detection of GLuc. We firstly constructed two plasmids pAAV2-neo-TTR-GLuc and pAAV2-neo-CMV-GLuc. Then, 4 cell lines were transfected with the two plasmids in aid of Lipofectamine 2000, including Huh7 and HepG2, which are derived from liver cells, as well as HEK293 and HeLaS3 cells, which are non-liver cell lines. We monitored the expression of GLuc in the supernatant of these cell cultures at different time points post-transfection. Furthermore, we injected the two plasmids with different doses into BALB/c mice by the means of hydrodynamic delivery and monitored the GLuc expression in vivo with 2.5 microl tail tip blood since 2 h post-injection. The cell assay results suggested that the expression of GLuc driven by CMV promoter was significantly higher than that of GLuc driven by TTR promoter. And, the luciferase activity of GLuc driven by CMV promoter was 50-300 times higher than that of GLuc driven by TTR promoter in HEK293 and HeLaS3 cell lines, but less than 10 times higher than that of GLuc driven by TTR promoter in the HepG2 and Huh7 cell lines, indicating the relative liver-specificity of TTR promoter. In the animal assay, the higher luciferase activity was determined in CMV promoter group than in TTR promoter group at different doses of the two plasmids. But the expression patterns for the two promoters differed obviously. The expression of GLuc driven by CMV promoter reached the maximum 10 hours post-injection and declined rapidly; while the expression of GLuc driven by TTR promoter reached the maximum 48 hours after delivery, and declined very slowly. These results implied that PTTR could keep expression of driven gene in a long time although its expression intensity is lower than PCMV's. Thus, it is more suitable for maintaining longer expression of target genes in liver.
Animals ; Cell Line ; Cytomegalovirus ; genetics ; metabolism ; Gene Expression ; Gene Transfer Techniques ; Genes, Reporter ; Humans ; Luciferases ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Prealbumin ; genetics ; metabolism ; Promoter Regions, Genetic

This study aimed to investigate the reconstruction of the thumb and finger extension function in patients with middle and lower trunk root avulsion injuries of the brachial plexus.From April 2010 to January 2015,we enrolled in this study 4 patients diagnosed with middle and lower trunk root avulsion injuries of the brachial plexus via imaging tests,electrophysiological examinations,and clinical confirmation.Muscular branches of the radial nerve,which innervate the supinator in the forearm,were transposed to the posterior interosseous nerve to reconstruct the thumb and finger extension function.Electrophysiological findings and muscle strength of the extensor pollicis longus and extensor digitorum communis,as well as the distance between the thumb tip and index finger tip,were monitored.All patients were followed up for 24 to 30 months,with an average of 27.5 months.Motor unit potentials (MUP) of the extensor digitorum communis appeared at an average of 3.8 months,while MUP of the extensor pollicis longus appeared at an average of 7 months.Compound muscle action potential (CMAP) appeared at an average of 9 months in the extensor digitorum communis,and 12 months in the extensor pollicis longus.Furthermore,the muscle strength of the extensor pollicis longus and extensor digitorum communis both reached grade Ⅲ at 21 months.Lastly,the average distance between the thumb tip and index finger tip was 8.8 cm at 21 months.In conclusion,for patients with middle and lower trunk injuries of the brachial plexus,transposition of the muscular branches of the radial nerve innervating the supinator to the posterior interosseous nerve for the reconstruction of thumb and finger extension function is practicable and feasible.

Objective To develop and evaluate a single -tube multiplex RT -real time PCR assay for the simultaneous detection of rotavirus,astrovirus and hepatitis A virus.Methods Gen -bank sequences of rotavirus,astrovirus and hepatitis A virus were included as reference sequences.Primers and probes were designed based on the reference sequences.A multiplex real -time RT -PCR assay was developed,and the reproducibility,specificity and sensitivity of the assay were evaluated.Fecal samples from 1 28 patients with viral diarrhea were detected and verified by gene sequencing.Results There was high reproducibility and specificity of the single -tube multiplex real -time RT -PCR assay for detecting rotavirus,astrovirus and hepatitis A virus.Detection limits of 1 01 copies were achieved in tests of rotavirus,1 02 copies for astrovirus and hepatitis A virus in one reaction.3.1 3% and 1 7.97% of 1 28 clinical specimens were detected positive for astrovirus RNA and rotavirus RNA respectively.And the positive samples were verified by gene sequencing.Conclusion Rotavirus,astrovirus and hepatitis A virus can be detected and identified by the single -tube multiplex RT -real time PCR assay with high specificity and sensitivity.The assay developed in this study can be applied to the clinical diagnosis and epidemiological investigation.

OBJECTIVEFive heavy metals (Hg, Cu, Cd, Pb, Cr) and arsenic (As) contents in 57 samples of from 19 localities in Sichuan, Qinghai and Gansu of Rhizoma et Radix Notopterygii sourcing from two plants Notoperygium incusum and N. forbesii has been analyzed to evaluate the content standard of heavy metal and arsenic for drafting the herbal medicine quality.

METHODICP-AES method was applied to determine As, Cu, Cd, Pb, Cr and CAAS method was applied to determine Hg.

RESULTHg can not be determined through CAAS, and little Pb was determined except in limited N. forbesii samples from certain sites, while the contents of Cu in most samples are higher than 20 mg x kg(-1). As in N. forbesii samples, and Cd in N. incisum samples are higher or approximate the limitation of related standard of medicinal plants and production. Cr varies from different localities, ranging from 0.48 to 8.36 mg x kg(-1) for N. incisum and 1.44-8.03 mg x kg(-1) for N. forbesii.

CONCLUSIONReferring to the related standards of medicine and vegetable, the heavy metals and arsenic contents of Rhizoma et Radix Notopterygii samples from traditional producing areas are confirmed the propositional standard, i. e., Pb < or = 5.0 mg x kg(-1), Cd < or = 0.3 mg x kg(-1), Hg < or = 0.2 mg x kg(-1), Cu < or = 28.0 mg x kg(-1), As < or = 2.0 mg x kg(-1), and Cr < or = 1.0 mg x kg(-1). Of these five heavy metals, Cu is much over the limitation standard, which suggested that GAP base of Rhizoma et Radix Notopterygii must be selected in lower Cu environmental condition in main producing area.

Apiaceae ; chemistry ; classification ; Arsenic ; analysis ; Cadmium ; analysis ; China ; Chromium ; analysis ; Copper ; analysis ; Ecosystem ; Metals, Heavy ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; classification ; Quality Control ; Rhizome ; chemistry