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 study the effects of the melt casting and the sintering methods on the machinability and other characteristics of K_2O-MgO-MgF_2-SiO_2 glass-ceramics. Methods: X-ray diffraction(XRD) and scanning electron microscopy (SEM)were used to examine the main crystal and the microstructure of the glass ceramics of the same composition which were made by melt casting and sintering methods, respectively. The transmissivity, flexural strength and brittleness index were observed respectively. Results: The glass ceramics made by two different methods have the same crystal, KMg_(2.5)Si_4O_(10)F_2, while the specimen prepared by the sintering method had lower transmissivity,better machinability and mechanical properties due to the higher mica volume percent. Conclusion; For production of fluoro-mica glass ceramics, sintering method has superiority compared to melt-casting method.

Objective Precondition is an approach to myocardial protection during ischemia-reper-fusion by inhibiting myocardial cell apoptosis.The purpose of this study was to evaluate the cardioprotective effect using 99Tcm-syuaptotagmin I (Syt I)-C2A to detect myocardial cell apoptosis in the myocardial is-chemia-repedusion rat model.Methods (1) The C2A domain of Syt I was labeled with 99Tcm using 2-iminothiophene hydrochloride (IT) method.Radiochemical purity was determined with thin layer chroma-tography.The binding activity of radiolabeled protein was assessed using eamptothecin-treated Jurket cells.(2) One group of 6 rats was prepared for myocardial ischemia-reperfusion model(A group),and another group of 6 rats was prepared for myocardial ischemia precondition model(B group).99Tcm-Syt I-C2A was injected via the tail vein at a dosage of about 7.4 MBq.At 1h after injection,the rat was sacrificed,and the heart was removed to rinse with saline and dye with triphenyl tetrazolium eoride (TTC).According to the resdt of myocardial dye,theischemic myoeardium was separated from the viable myocardium and weight was measured,and then its radioactivity was determined by gamma counting.The difference of radioactive uptake in the ischemic myocardium between these two group models was compared using percentage activity of injection dose per gram of tissue(%ID/g)±standard deviation(x±s).SPSS 12.0 was used for data analy-sis,and t-test was used to compare data.Results (1) The radiochemical purity of 99Tcm-Syt I-C2A was (98.90±0.43)%,and the radioactivity in the camptothecin-treated group was (10.99±0.55) folds higher than that of non-treated viable control group.(2)In the ischemia-reperfusion model,the radioactive uptake of 99Tcm-Syt,I-C2A was(2.41±0.32)%ID/g in the ischemic myocardium,and(0.16±O.02)%ID/g in the nomud myocardiunm.However,in the myocardial ischemia precondition model,(0.46±0.05)%ID/g in the isehemic myocardium was measured,and(0.20±0.05)%ID/g in the normal myocardium.Uptake of 99Tcm-Syt I-C2A in ischemic myocrdium showed statistically significant difference (t=8.52,P

OBJECTIVETo identify the affect of chronic low back pain on multifidus muscle atrophy and fatty infiltration.

METHODSFrom March 2010 to August 2013, a retrospective study were carried out in the department of orthopedics of patients with low back pain. Finally 31 cases were selected to this study including 19 males and 12 females with an average age of 36.4 years ranging from 23 to 55 years. The main symptoms of these patients were repeated back pain. Duration was more than 1 year. X-ray, CT, MRI showed no obvious abnormalities. The changes of net cross-sectional area of multifidus and T2 signal ratio of the same patient were measured at different time by MRI. VAS and Oswestry disability scores were recorded in two MRI examination. Correlation between these change of multifidus net area and T2 signal ratio in two times measurement and duration of low back pain, VAS, Oswestry disability scores were analyzed to find the affection of low back pain on paraspinal multifidus muscle.

RESULTSThe net multifidus cross-sectional area in same case by the second follow-up MRI is significantly smaller than that of the first follow-up, T2 signal ratio at second was significantly higher than that of the first (P < 0.05). The net cross sectional area of multifidus muscles reduced rate were positively correlated with VAS scores, duration and of Oswestry disabilitry scores (P < 0.001). The rate of increase in T2 signal ratio was not correlated with VAS scores,duration and the Oswestry disability scores (P > 0.05).

CONCLUSIONChronic low back pain is one of the most important reasons of paraspinal multifidus muscle atrophy and fatty. The duration, VAS and Oswestry disability scores of chronic low back pain were positively correlated with the multifidus muscle atrophy.

Adult ; Chronic Disease ; Female ; Humans ; Low Back Pain ; complications ; Male ; Middle Aged ; Muscular Atrophy ; diagnostic imaging ; etiology ; Paraspinal Muscles ; diagnostic imaging ; Radiography ; Retrospective Studies ; Young Adult

Objective To observe whether the expression of interferon-regulatory factor genes are re- lated to systemic lupus erythematosus (SLE).Methods The clinical data of 45 SLE patients and 37 normal controls were collected.Total RNA of peripheral blood was extracted and transcripted into cDNA.Sybr green dye based real-time quantitative PCR method was used to compare the expression (indicated as-??Ct value) of IRFI,IRF4,IRF8 in patients with SLE and those in the controls.Results The levels of IRF1,IRF4 and IRF8 mRNA were-3.90?0.19,-8.04?0.25 and 3.60?0.15 respectively in normal controls.In SLE patients, IRF4 mRNA expression was -8.82?0.18,higher than that in normal (P=0.011).But IRF8 mRNA expression was 3.09?0.13,lower than that in normal (P=0.012).Conclusion Abnormal IRF mRNA expression is found in the peripheral blood of SLE patients.IRFs may play roles in the pathogenesis of SLE by affecting the differen- tiation of Th cells.

BACKGROUNDErythropoietin (EPO) functions as a tissue-protective cytokine in addition to its crucial hormonal role in red cell production and neuron protection. This study aimed to determine the neuron protective effect of erythropoietin on experimental rats enduring spinal cord injury (SCI) by assessing thrombospondin-1 (TSP-1) level and transforming growth factor-beta (TGF-beta) in the development of a rat model of SCI.

METHODSSixty Sprague-Dawley rats were randomly assigned to three groups: sham operation control group, SCI group and EPO treatment group. By using a weight-drop contusion SCI model, the rats in the SCI group and EPO treatment group were sacrificed at 24 hours and 7 days subsequently. The Basso, Beattie, and Bresnahan (BBB) scores were examined for locomotor function. Pathological changes were observed after HE staining. The expressions of thrombospondin-2 (TSP-1) and TGF-beta were determined by immunohistochemical staining and Western blotting.

RESULTSSlighter locomotor dysfunction was discovered and it was recovered abruptly as higher BBB scores were found in the EPO treatment group than in the SCI group (P < 0.01). Pathologically, progressive disruption of the dorsal white matter and regeneration of a few neurons were also observed in SCI rats. TSP-1 and TGF-beta expression increased at 24 hours and 7 days after SCI in the injured segment, and it was higher in the SCI group than in the EPO treatment group. Spinal cord samples from the animals demonstrated a TSP-1 optical density of 112.2 +/- 6.8 and TSP-1 positive cells of 5.7 +/- 1.3 respectively. After injury, the TSP-1 optical density and cell number increased to 287.2 +/- 14.3/mm(2) and 23.2 +/- 2.6/mm(2) at 24 hours and to 232.1 +/- 13.2/mm(2) and 15.2 +/- 2.3/mm(2) at 7 days respectively. When EPO treated rats compared with the SCI rats, the TSP-1 optical density and cell number decreased to 213.1 +/- 11.6/mm(2) and 11.9 +/- 1.6/mm(2) at 24 hours and to 189.9 +/- 10.5/mm(2) and 9.3 +/- 1.5/mm(2) at 7 days, respectively (P < 0.01). In the SCI rats, the TGF-beta optical density and positive neuron number were 291.4 +/- 15.2/mm(2) and 28.8 +/- 4.9/mm(2) at 24 hours and 259.1 +/- 12.3/mm(2) and 23.9 +/- 4.1/mm(2) at 7 days respectively. They decreased in the EPO treated rats to 222.8 +/- 11.9/mm(2) and 13.7 +/- 2.1/mm(2) at 24 hours and to 196.5 +/- 9.7/mm(2) and 8.7 +/- 2.2/mm(2) at 7 days (P < 0.01).

CONCLUSIONSIncreased expression of TSP-1 and TGF-beta can be found in the injured segment of the spinal cord at 24 hours and 7 days after injury. EPO treatment can effectively prevent pathological alterations from severe spinal cord injury by reduced expression of TSP-1 and TGF-beta.

Animals ; Blotting, Western ; Disease Models, Animal ; Erythropoietin ; therapeutic use ; Female ; Immunohistochemistry ; Neuroprotective Agents ; therapeutic use ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; drug therapy ; metabolism ; Thrombospondin 1 ; metabolism ; Transforming Growth Factor beta ; metabolism

OBJECTIVETo evaluate the clinical outcomes of minimally invasive posterior lumbar interbody fusion (PLIF) assisted by X-Tube system for the management of degenerative lumbar diseases.

METHODSA total of 31 patients, 17 male and 14 female with ages from 38 to 75 (average 54.2 years), underwent minimally invasive PLIF assisted by the X-Tube system from May 2005 to March 2006. The index diagnosis was lumbar spondylolisthesis in 14 cases, spinal stenosis in 8 cases, separation of the posterior ring apophysis in 5 cases, intervertebral space stenosis with disk herniation in 4 cases. Before operation, conservative management for at least 6 months was proved to be failure in all these patients. The operative duration and blood loss were estimated . The changes of postoperative serum level of creatinine kinase was measured as well, and compared with the control group of 31 cases who were managed with traditional open PLIF operation during the same period at our department. The clinical functional outcomes were evaluated according to Oswestry disability questionnaire.

RESULTSThe operation lasted for 140-225 min, with a mean duration of (176 +/- 22) min. Blood loss during the operation was 270-750 ml, with a mean of (406 +/- 96) ml. Postoperative serum level of creatinine kinase was obviously lower in minimally invasive PLIF cases than in the open control cases. Although 2 pedicle screws in 2 cases were not in ideal position, there was no nerve root irritation or fixation failure and hence no revision was required. One case with spinal stenosis complained of numbness in the area dominated by left L5 nerve root after operation, but the symptom was relieved within 2 weeks through conservative management. All the 31 patients were followed up for 7-17 months, with a mean duration of 12.2 months. Lumbar radiography, and three-dimensional CT reconstruction in some cases, was performed and revealed solid fusion of the surgical segments half a year after the operation. The average Oswestry scores decreased from preoperative 40.6 +/- 5.1 to 17.4 +/- 6.5 at the first postoperative day and to 9.5 +/- 4.0 at the final follow-up. The outcome of this operation were rated as excellent.

CONCLUSIONSMinimally invasive PLIF assisted by X-Tube system has the characteristics of less blood loss, tissue trauma and operative time, quick recovery and bony fusion. The short-term outcomes are excellent.

Adult ; Aged ; Endoscopy ; Female ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Spinal Fusion ; instrumentation ; methods

OBJECTIVETo establish chemical pattern recognition method for the identification and evaluation of Atractylodes macrocephala.

METHODThe chemical constituents in methanol extract of 32 samples of A. macrocephala were determined by HPLC. The fingerprints were obtained and were handled by hierarchical clustering analysis and stepwise discriminant analysis.

RESULT AND CONCLUSIONAccording to the result of classification, all samples collected were devided into three Grades--the superior, the ordinary and the fake. Chemical pattern recognition method was established. It may be of practical value for the quality control of A. macrocephala.

Atractylodes ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Cluster Analysis ; Pattern Recognition, Automated ; Plants, Medicinal ; chemistry ; Quality Control ; Rhizome ; chemistry

To evaluate the adjuvant effect of recombinant enterovirus 71 (EV71) subunit vaccine formulated with chitosan, rabbits were orally immunized with recombinant VP1 (rVP1) or rVP1 mixed with chitosan adjuvant. Levels of virus-specific IgG and IgA antibodies in sera, mucosal wash buffer (intestine, nasal cavity, and lung), and feces were determined by indirect enzyme-linked immunosorbent assay (ELISA). The titers of neutralizing antibodies against EV71 were determined using cytopathic effect-based neutralizing assay, and levels of cytokines (IFN-gamma and IL-4) secreted from in vitro-cultured rabbit splenic lymphocytes under antigen stimulation were also determined by ELISA. Results showed that immunization with rVP1 alone could only induce low levels of serum IgG and mucosal IgA, while rVP1 combined with chitosan adjuvant were able to induce significantly higher levels of antibodies, rVP1 can only induce neutralizing antibodies when used in combination with chitosan. Levels of IFN-gamma and IL-4 in the group immunized with rVP1 plus chitosan were significantly higher than those in the group immunized with rVP1 only or those in the control groups. Our study lays the foundation for development of oral VP1 vaccine against EV71 infection.
Adjuvants, Immunologic ; administration & dosage ; Animals ; Antibodies, Viral ; immunology ; Chitosan ; administration & dosage ; immunology ; Enterovirus A, Human ; genetics ; immunology ; Enterovirus Infections ; immunology ; prevention & control ; virology ; Female ; Humans ; Rabbits ; Vaccination ; Vaccines, Subunit ; administration & dosage ; genetics ; immunology ; Viral Proteins ; administration & dosage ; genetics ; immunology ; Viral Vaccines ; administration & dosage ; genetics ; immunology

Objective To investigate the characteristics of EGFR,ALK and ROS1 mutations in patients with non-small cell lung cancer (NSCLC) in South of China and its relationship with clinical features.Methods The tumor tissues and corresponding clinical data of 76 NSCLC patients in South of China from November 2016 to June 2017 were collected.The mutations of EGFR,ALK and ROS1 were detected by ARMS assay with Joint detection kit.Meanwhile,the correlation between gene mutation rate and clinical features was analyzed.Results The mutation rate of EGFR was 67.3％ (42/76) in 76 patients with NSCLC in South of China,19 del and L858R mutations were the main mutation types.There was a co-mutation including 19 del and L858R.The positive rate of ALK gene fusion was 17.1％ (13/76),and 4 cases of ALK gene fusion combined with EGFR mutation were detected.The positive rate of ROS1 gene fusion was 1.3％ (1/76),and there was no co-mutation with other genes.Compared with ROS1,EGFR and ALK mutation rate was higher,the difference was statistically significant (x2 =54.515,P =0.000;x2 =11.329,P =0.001).The mutation rate of EGFR in non-smoking NSCLC patients was significantly higher than that in smokers (x2 =4.578,P=0.032),while the mutation rate of ALK and ROS1 was not statistically significant (x2=0.000,P＞0.05).There was no statistically significant difference in EGFR,ALK and ROS1 gene mutation rates among NSCLC patients of different age,sex and histology (x2 =0.000 ～ 2.219,P＞ 0.05).Conclusion EGFR,ALK and ROS1 gene mutations can be seen in patients with NSCLC in South China,in which EGFR and ALK gene mutation rate is higher.