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

OBJECTIVETo compare the differences between computer aided identification and ultra-thin layer slicing in measuring the lesions of femoral head necrosis,and to confirm the accuracy and practicability of computer aided method.

METHODSFrom June 2012 to December 2013, the X-ray and MRI of 24 patients (24 hips on unilateral) were reviewed, who had avascular necrosis of the femoral head (ANFA) at late stage (stage III and IV) according to the ARCO international staging system. There were 15 males and 9 females, with an average age of (65.1 ± 8.8) years old, ranged 33 to 74 years old. Based on the software system with seeds point identification, the ragional adaptive search method with computer aid was used to calculate the volume of necrotic lesion in femoral on MRI. Then the pathological slices of those intraoperative femoral heads were made to measure the gross volume of necrotic lesion in femoral head,and the values were compared with the data in the computer.

RESULTSFor 24 hips, by the calculation of computer, the necrotic volume was (20.00 ± 3.04) cm (ranged, 18.72 to 21.29 cm³). Under the pathological section, the necrotic volume of the femoral head was (19.89 ± 3.17) cm³ (ranged, 18.55 to 21.23 cm³). In computer and pathology two kinds of measurement, the two entire femoral head volume had no significant difference using these two measurements (t = -1.227, P = 0.232).

CONCLUSIONComputer aided identification for necrotic area of femoral head adaptive can demonstrate the morphology of femoral head necrosis accurately and reliably, which will help surgeon better understand the morphology and orientation in femoral head.

Adult ; Aged ; Diagnosis, Computer-Assisted ; Female ; Femur Head Necrosis ; pathology ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged

Background There are a lot of studies about the carrier of corneal endothelial transplantation,but the best carrier has not been defined.Objective This study was to investigate the biocompatibility of chitosan carrier with rabbit corneal endothelium in vivo.Methods Fresh eye-balls were obtained from 10 New Zealand white rabbits.Rabbit corneal endothelial cells (CECs) were isolated and cultured on chitosan carrier in vitro.The morphology and density of rabbits CECs were observed every day,and the expressions of fibronectin (FN),collagen-1 (Coil-I) and Zonula occludens 1 (ZO-1) were detected by immunoinfluorescence.The morphology and ultrastructure of CECs were observed under the scanning and transmission electron microscope.Chitosan carrier with CECs was implanted into the anterior chamber of the left eyes in ten healthy New Zealand white rabbits,and only paracentesis of anterior chamber was performed in the right eyes as controls.The inflammation of ocular anterior segment was examined under the slit lamp microscope,and corneal thickness was measured 1 week,4 and 8 weeks after operation.Corneal endothelium cell density and morphology were examined under the corneal endothelial microscope at postoperative 2 weeks.Corneal samples were collected for the regular histopathological examination to observe the inflammatory reaction at postoperative 1 month and 3 months.Paired t test was used for statistical analyses between the control group (left eyes) and the experimental group (right eyes).The use and care of the animals followed the Statement of ARVO.Results CECs formed an intact monolayer of cells with the uniform shape and size on the chitosan membrane after incubated for 5 days.The cells reached confluence of 90％ 7 days after cultured with the 40％ hexagon cells.Under the scanning electron nicroscope,rabbit CECs showed the round or polygon in the shape with the microvillus on the cell surface.The cells connected closely by desmosome.The processes,pseudopodiums and microvillus on the cellular surface,vacuole in the cytoplasm,expanded endoplasmic reticulum with ribosome and abundant chromatin were exhibited under the transmission electron microscope.The immunofluorescence examination revealed the positive expressions of FN,Coll-Ⅰ and ZO-1 in the CECs on the chitosan carrier.In the in vivo experiment,the exudation in the anterior chamber and corneal edema were seen under the slit lamp microscope 3 days after implantation of chitosan carrier with CECs.However,the inflammation was gone 14 days after operation.The differences of the corneal thickness were no significant between the experimental group and the control group 1 week and 4,8 weeks after operation (t =1.377,P=0.265;t =1.795,P=0.165 ; t =0.390,P =0.760).In addition,no significant differences were found in the CECs density and the hexagon cells rate between the two groups(P =0.365,0.062).The histopathological examination showed that the inflammatory cells around the chitosan membrane were disappeared 3 months after operation and showed a good corneal structure.Conclusions Chitosan carrier has a good biocompatibility with rabbit CECs and anterior chamber,and it may be a potentially good carrier for CECs transplantation.

OBJECTIVETo compare the biomechanical performances of different wires and cable fixation devices in posterior instrumentation for atlantoaxial instability, and test the effect of different fixation strengths and fixation approaches on the surgical outcomes.

METHODSSix specimens of the atlantoaxial complex (C0-C3) were used to establish models of the normal complex, unstable complex (type II odontoid fracture) and fixed complex. On the wd-5 mechanical testing machine, the parameters including the strength and rigidity of anti-rotation, change and strength of stress, and stability were measured for the normal complex, atlantoaxial instability complex, the new type titanium cable fixation system, Atlas titanium cable, Songer titanium cable, and stainless wire.

RESULTSThe strength and rigidity of anti-rotation, change and strength of stress, stability of flexion, extension and lateral bending of the unstable atlantoaxial complex fixed by the new double locking titanium cable fixation system were superior to those of the Songer or Atlas titanium cable (P<0.05) and medical stainless wire (P<0.05). Simultaneous cable fastening on both sides resulted in better fixation effect than successive cable fastening (P<0.05). Better fixation effect was achieved by fastening the specimen following a rest (P<0.05).

CONCLUSIONSThe fixation effects can be enhanced by increased fastening strengths. The new type double locking titanium cable fixation system has better biomechanical performance than the conventional Songer and Atlas titanium cables. Fastening the unstable specimens after a rest following simultaneous fastening of the specimen on both sides produces better fixation effect.

Atlanto-Axial Joint ; physiopathology ; surgery ; Biomechanical Phenomena ; Bone Wires ; Cadaver ; Humans ; Internal Fixators ; Joint Instability ; physiopathology ; surgery ; Orthopedic Fixation Devices

ObjectiveTo analyze Echinococcus infection in definitive and intermediate hosts in different zones of Qinghai plateau,Qinghai southern plateau,Qilian mountain-Hehuang valley and Chaidamu basin,and to provideascientificbasisfor developing controlstrategiesagainstEchinococcosisinfection. Methods Echinococcosis infection in definitive hosts,dogs and foxes,was identified by morphological observation; in domesticated and wild intermediate host animals was identified by anatomy and pathology; some of the suspected samples were further identified by molecular biological methods.ResultsStray dogs in different zones of Qinghai plateau were infected with Echinococcus granulosus,the infection rates were 38.71％(300/775),49.60％(124/250),and 9.76％(4/41 ) in Qinghai southem plateau,Qilian mountain-Hehuang valley and Chaidamu basin,respectively,and the difference was statistically significant(x2 =25.72,P ＜ 0.01 ).in addition,only Qinghai southern plateau dogs were infected with Echinococcus multiloularis,and the infection rate was 16.04％(98/611).The infection rates of fox with Echinococcus multilocularis were 22.89％(38/166) and 30.77％(12/39) in Qinghai southern plateau and Qilian mountain-Hehuang valley,respectively,and wolves were also found to be infected with Echinococcus granulosus in the same areas.The infection rates of domesticated sheep,yaks,goats and pigs with Echinococcosis were significantly different statistically in those different areas(x2 =82.70,41.82,212.63,194.58,all P ＜ 0.01 ).The infection rates of sheep and yaks were higher[43.43％(5664/13 042),49.47％(2917/5896),52.99％ (887/1674),42.18％ (779/1847),50.70％ (1049/2069),52.90％ (685/1295) ] in three areas.The infection rates of goats and pigs [3.26％ (7/215),0.00％ (0/108)] in Qinghai southern plateau were lower than that of other two areas[ 19.51％(119/610),26.91％(43/1598),47.91％(343/716),21.91％(71/324)].The infection rates of Ochotona curzoniae with Echinococcosis were 6.21％ (243/3910),1.80％ (3/167) and 0.00％ (0/199) in Qinghai southern plateau,Qilian mountain-Hehuang valley and Chaidamu basin,respectively,and the difference was statistically significant (x2 =18.50,P ＜ 0.01 ).Moreover,wild intermediate hosts of Echinococcosis,such as Microtus fuscus,Lepus oiostolus,Pseudois nayaur,Procapra picticaudata,and Prodorcas gutturosa were found to be infected only in Qinghai southern plateau.ConclusionsHuman is faced with a threat of Echinococcosis infection from various definitive hosts in different zones of Qinghai plateau.And stray dogs are the most crucial factor.The life-cycles of Echinococcus are very complicated in Qinghai plateau.Qinghai plateau is a key area in prevention and control of Echinococcosis infection in China.

A LC-MS method was established for the determination of the protein binding rates of oleanolic acid in human plasma and serum albumin. The equilibrium dialysis combined with LC-MS to determine the total concentration in plasma and free drug concentration of oleanolic acid was carried out. The human plasma protein binding rates of oleanolic acid at three concentrations were 79.6%, 81.9% and 63.3%, respectively. The human serum albumin protein binding rates of oleanolic acid at three concentrations were 53.5%, 56.6% and 47.7%, respectively. The method is shown to be simple, accurate, sensitive and specific for the determination of biological samples. The protein binding rates in human plasma and serum albumin were of high strength.
Chromatography, Liquid ; methods ; Dialysis ; Humans ; Mass Spectrometry ; methods ; Oleanolic Acid ; blood ; Protein Binding ; Sensitivity and Specificity ; Serum Albumin ; metabolism

Objective To study the correlation between the deformation and displacement trend of the lumbosacral vertebra (L1-S1) for two typical scoliosis spines under vertical loads,so as to provide the mechanical basis of treatment and prevention of scoliosis in clinic.Methods The X-ray computed tomography (CT) images of two typical scoliosis spines (Lenke-4AN type and Lenke-5CN type) were converted into 3D models,and their finite element models were then established and verified.The internal stress distribution and displacement variation of the models were calculated by the finite element software;the correlation between the lumbosacral vertebral structure and displacement of the spine was analyzed.Results Under the same boundary conditions and load cases,the stress and displacement for two kinds of lumbosacral vertebral models showed different trends.Due to its leftleaning and forward convex bending deformation as well as the relatively large lordosis angle (60°) and smaller left-leaning angle (17.37°),the Lenke-4AN type lumbosacral spine produced slightly small forward convex displacement (8.18 mm) and relatively large left-leaning displacement (0.97 mm).The Lenke-5CN type lumbosacral spine showed left-leaning and forward convex bending deformation as well,with relatively large lordosis angle(59°) and left-leaning angle (26.97°),so it produced more severe left-leaning displacement (20.65 mm) andforward convex displacement (9.22 mm).Conclusions The deformation trend of lumbosacral vertebra is closelyrelated to its structural characteristics,and different scoliosis lumbosacral vertebral structures will cause the corre-sponding deformation trend.The research findings are important for the prevention and treatment of scoliosis.

OBJECTIVETo observe effect of Xue Hanjing oral fluid on mice immunological function.

METHODThe weight of thymus gland and spleen and the function of abdominal cavity macrophage were measured. Production of the hemolysin antibody, the immunoglobulin of blood serum and complement and the proliferation of T lymphocytes were observed respectively by means of microblood, immune-turbidimetry and MTT staining.

RESULTXue Hanjing oral fluid could enhance index of the thymus gland and spleen and the phago-percent of abdominal cavity macrophage, increase the immunoglobulin of blood serum(IgG and IgM), and accelerate production of the hemolysin antibody and the proliferation of T lymphocytes.

CONCLUSIONSXue Hanjing oral can reinforce immunological function in mice.

Adjuvants, Immunologic ; pharmacology ; Administration, Oral ; Animals ; Antibody Formation ; Cell Division ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Female ; Hemolysin Proteins ; immunology ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Macrophages, Peritoneal ; physiology ; Male ; Mice ; Organ Size ; drug effects ; Phagocytosis ; drug effects ; Plants, Medicinal ; chemistry ; Poaceae ; chemistry ; Random Allocation ; T-Lymphocytes ; cytology

This study was aimed to investigate the clinico-pathological features, diagnosis and treatment of the 8p11 (eight p11) myeloproliferative syndrome (EMS). Morphological changes of cells were evaluated by bone marrow smear and biopsy. The cell immunophenotypes were analysed by flow cytometry. Karyotypes were determined by conventional cytogenetic method, and bcr/abl fusion gene was detected by reverse transcription-polymerase chain reaction (RT-PCR). The results indicated that EMS was a relatively rare disease characterized by the occurrence of a bcr/abl-negative myeloproliferative disorder and a T-cell lymphoblastic lymphoma (T-LBL). Bone marrow examination showed myeloid hyperplasia or myeloproliferative neoplasm, often accompanied by eosinophilia. Flow cytometric immunophenotyping showed increased myelomonoblasts; cytogenetic analysis showed a translocation at the 8p11 locus; RT-PCR demonstrated non bcr/abl fusion gene. At the molecular level, all cases carried a chromosomal abnormality involving the fibroblast growth factor receptor 1 (FGFR1) at chromosome 8p11. Up to now, 11 partner genes have been identified and associated with FGFR1 rearrangements. The most common partner is ZNF198 on chromosome 13q11-12. Majority of patients terminate in acute myeloid leukemia which is resistant to conventional chemotherapy. Currently, the only curative option appears to be allogeneic hematopoietic stem cell transplantation. In conclusion, EMS is myeloid and lymphoid neoplasm, associates with FGFR1 rearrangements. It is usually misdiagnosed as T-LBL, atypical chronic myeloid leukemia (aCML) or chronic myelogenous-monocytic leukemia (CMML). Timely cytogenetic and molecular biological examination is vital in order to avoid misdiagnosis and mistreatment.
Bone Marrow Cells ; pathology ; Chromosomes, Human, Pair 8 ; genetics ; Humans ; Male ; Middle Aged ; Myeloproliferative Disorders ; pathology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; pathology ; Receptor, Fibroblast Growth Factor, Type 1 ; genetics

Amyloid β protein (Aβ) is closely involved in the pathogenesis of Alzheimer's disease (AD), and one of the main strategies for AD treatment is antagonizing the neurotoxicity of Aβ or even clearing the Aβ deposited in the brain. The present study was aimed to observe the effects of intrahippocampal injection of Aβ₃₁₋₃₅ on the spatial learning and memory of rats by using Morris water maze technique, and explore the neuroprotective effects and possible mechanism of [Gly14]-humanin (HNG) against Aβ-induced deficits in learning behavior. The results showed that bilateral intrahippocampal injection of 2.0 nmol Aβ₃₁₋₃₅ significantly increased the mean traveled distance of rats in searching for the hidden underwater platform and decreased the distance percentage in the target quadrant in probe test after withdrawal of platform, whereas pretreatment with HNG (0.2 nmol and 2.0 nmol) suppressed Aβ₃₁₋₃₅-induced increase in the traveled distance and decrease in swimming distance percentage. Application of Genistein (40 nmol), a specific tyrosine kinase inhibitor, almost completely blocked the antagonistic effects of HNG against Aβ₃₁₋₃₅. These results indicate that HNG can dose-dependently prevent against Aβ₃₁₋₃₅-induced impairment in spatial learning and memory of rats, and the neuroprotective effects of HNG might involve the activation of endogenous tyrosine kinase pathway, suggesting that up-regulation of the tyrosine kinase signaling by using HNG might be of great significance for the improvement of cognitive function in AD.
Alzheimer Disease ; physiopathology ; Amyloid beta-Peptides ; adverse effects ; antagonists & inhibitors ; Animals ; Brain ; drug effects ; Genistein ; pharmacology ; Memory ; drug effects ; Neuroprotective Agents ; pharmacology ; Peptide Fragments ; adverse effects ; antagonists & inhibitors ; Peptides ; pharmacology ; Rats ; Spatial Learning ; drug effects