Objective To establish a loop-mediated isothermal amplification(LAMP) quantitative method for rapid detection of Japanese encephalitis and dengue fever virus.Methods According to the LAMP principle,design primers for LAMP detection and reaction system,establish LAMP detection method,and to evaluate the linear relationship between initial copy number and the specificity,sensitivity,repeatability and the reaction time(fluorescence signal value of 1×104 corresponding time).Results One sets of LAMP primers could be used to complete the detection work in 0.5 h.The sensitivity of LAMP detection technology was 10 times higher than that of the classical PCR technology,and no cross reaction with other viruses,and the coefficient of variation of the average test was less than 5%.There was a good linear relationship between cycle threshold and template concentration.Conclusion This method has high specificity,sensitivity,simple operation,which is easy to get the results,low equipment requirements and rapid,suitable for primary health institutions and the field inspection agencies for wide applications.

ObjectiveTo explore the clinical therapeutic effect and safety of treating the vertebral artery type cervical spondylosis with rotational manipulation.Methods180 patients with vertebral artery type cervical spondylosis were treated by rotational manipulation.ResultsBy follow-up, 89 patients were evaluated as excellent, 73 as good and 18 as fair. The total rate of excellent and good results was 81%.ConclusionRotational manipulation is an effective method. The correct diagnosis and indication are keys to take good treatment results and safety.

ObjectiveTo develop a stable model of focal cerebral infarction in rat to study the curative effect of neural stem cells transplantation.MethodsThirty-seven rats were selected which were divided into two groups in random, experimental group and control group. The focal infarction model was developed by the ligation of the left middle cerebral artery followed by the ligation of the ipsilateral common carotid artery and the temporary clip occlusion of the contralateral common carotid artery for 1.5 h. The operation adopted minimally invasive craniotomy though temporal bone. The model was evaluated by examining the neurologic deficits, ink perfusion, TTC staining and Magnetic Resonance imaging.ResultsAll the rats were in good condition after the operation, the mortality rate was 6.25% after 4 weeks. Ink perfusion and TTC staining confirmed that the ischemia was confined to the cortex. The areas of infarction measured 83.52 mm3 by Magnetic Resonance imaging after 4 weeks.ConclusionA stable focal cerebral infarction model can be achieved by minimally invasive craniotomy. It is superior for its homogeneity of infarction volume and site, and its low mortality. It can be used for the study of transplantation of neural stem cells.

Background:Traditional tissue engineering methods to fabricate urinary tract patch have some drawbacks such as compromised cell viability and uneven cell distribution within scaffold.In this study,we combined three-dimensional (3D) bioprinting and tissue engineering method to form a tissue-engineered urinary tract patch,which could be employed for the application on Beagles urinary tract defect mode to verify its effectiveness on urinary tract reconstruction.Methods:Human adipose-derived stem cells (hADSCs) were dropped into smooth muscle differentiation medium to generate induced microtissues (ID-MTs),flow cytometry was utilized to detect the positive percentage for CD44,CD105,CD45,and CD34 of hADSCs.Expression of vascular endothelial growth factor A (VEGFA) and tumor necrosis factor-stimulated gene-6 (TSG-6) in hADSCs and MTs were identified by Western blotting.Then the ID-MTs were employed for 3D bioprinting.The bioprinted structure was encapsulated by transplantation into the subcutaneous tissue of nude mice for 1 week.After retrieval of the encapsulated structure,hematoxylin and eosin and Masson's trichrome staining were performed to demonstrate the morphology and reveal collagen and smooth muscle fibers,integral optical density (IOD) and area of interest were calculated for further semiquantitative analysis.Immunofluorescent double staining of CD31 and a-smooth muscle actin (α-SMA) were used to reveal vascularization of the encapsulated structure.Immunohistochemistry was performed to evaluate the expression of interleukin-2 (IL-2),α-SMA,and smoothelin of the MTs in the implanted structure.Afterward,the encapsulated structure was seeded with human urothelial cells.Immunofluorescent staining of cytokeradns AE1/AE3 was applied to inspect the morphology of seeded encapsulated structure.Results:The semi-quantitative assay showed that the relative protein expression of VEGFA was 0.355 ± 0.038 in the hADSCs vs.0.649 ± 0.150 in the MTs (t--3.291,P =0.030),while TSG-6 expression was 0.492 ± 0.092 in the hADSCs vs.1.256 ± 0.401 in the MTs (t =3.216,P =0.032).The semi-quantitative analysis showed that the mean IOD of IL-2 in the MT group was 7.67 ± 1.26,while 12.6 ± 4.79 in the hADSCs group,but semi-quantitative analysis showed that there was no statistical significance in the difference between the two groups (t =1.724,P =0.16).The semi-quantitative analysis showed that IOD was 71.7 ± 14.2 in noninduced MTs (NI-MTs) vs.35.7 ± 11.4 in ID-MTs for collagen fibers (t =3.428,P =0.027) and 12.8 ± 1.9 in NI-MTs vs.30.6 ± 8.9 in ID-MTs for smooth muscle fibers (t=3.369,P=0.028);furthermore,the mean IOD was 0.0613 ±0.0172 in ID-MTs vs.0.0017 ± 0.0009 in NI-MTs for α-SMA (t =5.994,P =0.027),while 0.0355 ± 0.0128 in ID-MTs vs.0.0035 ± 0.0022 in NI-MTs for smoothelin (t=4.268,P =0.013),which indicate that 3D bioprinted structure containing ID-MTs could mimic the smooth muscle layer of native urinary tract.After encapsulation of the urinary tract patch for additional cell adhesion,urothelial cells were seeded onto the encapsulated structures,and a monolayer urothelial cell was observed.Conclusion:Through 3D bioprinting and tissue engineering methods,we provided a promising way to fabricate tissue-engineered urinary tract patch for further investigation.

Objective:To investigate the impacts of galangin(Gal)on the immune inflammatory response in preeclampsia(PE)rats by regulating the IL-6/signal transducer and activator of transcription 3(STAT3)signaling pathway.Methods:Pregnant rats were injected subcutaneously with L-arginine methyl ester(L-NAME 125 mg/kg)to establish a PE model.After successful modeling,they were randomly grouped into Model group,low,medium and high doses Gal groups(10,30,and 60 mg/kg)and activator group(60 mg/kg Gal+0.05 mg/kg IL-6/STAT3 pathway activator rIL-6).Twelve pregnant rats of the same period were randomly selected as the control group,and each group was given corresponding drugs by gavage and intraperitoneal injection,once a day for a continuous week.The levels of mean caudal artery pressure(MAP)and 24-hour urinary protein and blood urea nitrogen(BUN)in rats during the third trimester of pregnancy were measured using a blood pressure tester and a fully automated biochemical analyzer;the number and weight of the average offspring of pregnant mice in each group were calculated,and the pregnancy outcome was analyzed;HE staining was applied to observe the pathological damage of rat placenta;ELISA was applied to detect the levels of IL-6,IL-1β and TNF-α in the placenta of rats in each group;the proportion of T lymphocytes in rat peripheral blood was detected by flow cytometry;Western blot was applied to detect the expression of IL-6/STAT3 pathway protein in rat placenta.Results:Compared with control group,the lev-els of MAP,24-hour urinary protein,BUN,IL-6,IL-1β,TNF-α,the CD8+T cells,and the expression levels of IL-6 and p-STAT3/STAT3 proteins in Model group were obviously increased,the placenta tissue was severely damaged,the mean offspring number and mean offspring weight,the proportions of CD3+T,CD4+T,and CD4+/CD8+T cells were decreased(P<0.05);compared with Model group,the levels of MAP,24-hour urinary protein,BUN,IL-6,IL-1β,TNF-α,the CD8+T cells,and the expression levels of IL-6 and p-STAT3/STAT3 proteins in Gal low,medium and high doses groups were obviously decreased,the damage to placental tissue was reduced,the mean offspring number and mean offspring weight,the proportions of CD3+T,CD4+T,and CD4+/CD8+T cells were in-creased(P<0.05);compared with Gal high-dose group,the levels of MAP,24-hour urinary protein,BUN,IL-6,IL-1β,TNF-α,the CD8+T cells,and the expression levels of IL-6 and p-STAT3/STAT3 proteins in activator group were obviously increased,the placenta tissue was aggravated,the mean offspring number and mean offspring weight,the proportions of CD3+T,CD4+T and CD4+/CD8+T cells were decreased(P<0.05).Conclusion:Gal may play a role in alleviating PE symptoms and improving pregnancy outcomes by inhibit-ing the IL-6/STAT3 pathway,inhibiting inflammatory reactions,and regulating cellular immunity.

BACKGROUND AND OBJECTIVES: Imidapril, one of the ACE inhibitor, has been used to treate hypertension, congestive heart failure, diabetes mellitus and renal disease. ACE inhibitor, however often causes dry cough and this adverse effect affects the compliance rate negatively. This report aimed to examine the incidence rate of coughing caused by Imidapril treat-ment. And we compared the results with the incidence and tendency of dry cough caused by the other ACE inhibitors. MATERIALS AND METHOD: This study enrolled 38 patients who followed up at internal medicine and took Imidapril from the period of May 9, 1997 to December 17, 1997. We tested its effectiveness, safety and tolerance. The initial dosage of it was 5mg/day and the maximum dosage according to blood pressure was 10mg/day for 8 weeks. The tendency and the incidence of dry cough have been analyzed and compared with recently published Korean reports. RESULTS: The 5 mg dosage of Imidapril showed 68.1% in lowering blood pressure rate and it went up to 81.8% during the 8 weeks treatment perriod. The blood pressure regularity rate was 36.3% after the 8 weeks treatment. The overall rate of adverse effect occurrence was 26.1% (6/23) and 8.7% of them were likely to cough. One of the patients who showed nausea stopped medication. The recent articles reported that the incidence of dry cough had been reduced: Enalapril (33.8%), Fosinopril (16%), Ramipril (15%), Imidapril (7.8 - 8.7%), Losartan (4.3%). CONCLUSION: Imidapril is as safe and effective as other ACE inhibitors. Various ACE inhibitors that were newly developed show their effectiveness in lessening incidence rate of dry cough, and Imidapril is thought to be an excellent drug for this matter. Even though Losartan shows the lowest incidence rate of dry cough, it needs to more regular usages since it is still on the stage of its medical adaptation.
Angiotensin-Converting Enzyme Inhibitors ; Angiotensins* ; Blood Pressure ; Compliance ; Cough* ; Diabetes Mellitus ; Enalapril ; Fosinopril ; Heart Failure ; Humans ; Hypertension ; Incidence* ; Internal Medicine ; Losartan ; Nausea ; Peptidyl-Dipeptidase A* ; Ramipril

OBJECTIVETo evaluate the effect of perioperative continuous epidural morphine administration on plasma D-dimer level in patients undergoing total hip replacement.

METHODSForty ASA I-II patients undergoing total hip replacement under epidural anesthesia were randomized into two groups. In one group, the patients were given epidural administration of morphine 15 min before operation at 4 mg (in 10 ml normal saline) and for 48 h after the operation at 80 microg/h, while those in the other group received epidural injection of the same amount of normal saline before operation and 0.15% ropivacaine 2.0 ml/h for 48 h in the same manner after operation. Blood samples were taken before anesthesia (T(0)), at the end of operation (T(1)), and 24 h and 48 h after operation (T(2) and T(3)) for determination of plasma IL-6 and D-dimer levels.

RESULTSIn both groups plasma IL-6 and D-dimer levels showed significant increase at T(1), T(2) and T(3) in comparison with those at T(0), and their levels were significantly lower in morphine group than in ropivacaine group at T(1), T(2) and T(3).

CONCLUSIONEpidural morphine can lower plasma IL-6 and D-dimer levels and correct blood hypercoagulability in patients undergoing total hip replacement.

Aged ; Arthroplasty, Replacement, Hip ; Female ; Fibrin Fibrinogen Degradation Products ; metabolism ; Humans ; Injections, Epidural ; Interleukin-6 ; blood ; Male ; Middle Aged ; Morphine ; administration & dosage ; Perioperative Care ; Postoperative Period

OBJECTIVETo assess the necessity of routine prophylactic irradiation at the level Ib for nasopharyngeal carcinoma (NPC).

METHODSNewly diagnosed NPC patients between January, 2001 and June, 2005 were enrolled in this study. The nodal distribution in each region was calculated from the data of transversal contrast enhance CT or magnetic resonance scan of the head and neck.

RESULTSCervical node involvement was found in 75.1% of the 338 patients enrolled. The rates of involvement at levels Ib, IIa, IIb, III, IV, Va, Vb and in the supra-clavicular region were 0.9%, 49.1%, 60.7%, 26.0%, 5.9%, 9.5%, 3.8% and 0.9%, respectively. Skip metastasis occurred only in 2.4% of the cases. The high risk region (defined by a probable risk>5%) of nodal metastases was (1) the ipsilateral levels III, IV, Va, and Vb in case of level II involvement, (2) the ipsilateral levels II, IV, Va, and Vb in case of level III involvement, (3) the ipsilateral levels II, III, Va, Vb and the supra-clavicular region in case of level IV involvement, (4) the ipsilateral levels II, III, IV, Vb and the supra-clavicular region in case of level Va involvement, (5) the ipsilateral levels II, III, IV, Vb, and the supra-clavicular region in case of level Vb involvement, (6) the contralateral levels II, III, and Va in case of unilateral cervical node involvement.

CONCLUSIONNodal involvement in NPC patients rarely occurs at the level Ib, which is not a high risk region whatever the regions may be to have lymph node metastasis and therefore does not need routine prophylactic irradiation.

Adolescent ; Adult ; Aged ; Female ; Humans ; Lymphatic Metastasis ; pathology ; radiotherapy ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; radiotherapy ; Neck ; Neoplasm Staging ; Tomography, X-Ray Computed ; Young Adult

Five structural important residues of rice nonspecific lipid transfer protein LTP110 were mutated by site-directed mutagenesis. Sequence results showed that they were all mutated successfully. After trying various E. coli expression systems, thioredoxin fusion expression system was found to be a proper system to express wild type and mutant LTP110. cDNA sequences encoding wild type LTP110 and the mutants Y17A, P72L, R46A, D43A, C50A were cloned into two kinds of thioredoxin fusion expression vectors. The expression results were compared. In pTrxFus/GI724 expression system, wild type LTP110 and the mutants Y17A, P72L, R46A could be expressed at low level while D43A and C50A could not be expressed normally; in pET32a(+)/BL21 (DE3) trxB- expression system, wild type LTP110 and all mutant proteins could be expressed very well and the levels were higher than that in pTrxFus/GI724 system. LTP110 fusion protein expressed in pET32a(+) vector was purified and its activity was checked by fluorescence labeled fatty acid. Results indicated that the recombinant LTP110 fusion protein has lipid binding activity. This work provides good basis for the further study.
Amino Acid Sequence ; Carrier Proteins ; genetics ; isolation & purification ; metabolism ; Gene Expression ; Genetic Engineering ; Genetic Vectors ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Oryza ; genetics ; Plant Proteins ; genetics ; isolation & purification ; metabolism ; Recombinant Fusion Proteins ; genetics ; isolation & purification ; metabolism ; Thioredoxins ; genetics

Objective To investigate the effects of standardized tertiary rehabilitation (STR) on limb motor function (LMF) after stroke.Methods Eighty-two patients were divided into a primary cerebral infarction group (PCI group) and a primary cerebral hemorrhage group (PCH group),and then randomly further divided into experi- mental and control sub-groups.All patients received routine internal medicine treatment,supplemented with stand- ardized tertiary" rehabilitation in the experimental groups.All patients were assessed with the simplified Fugl-Meyer motor function assessment (S-FMMFA) at enrollment,and 1,3 and 6 months after their stroke.Results The scores of the experimental groups were higher than those of the controls.The experimental groups scores were 26.10% of normal at the time of the enrollment,and improved to 42.52%,65.62% and 83.71% by the end of the 1st,3rd and 6th month,respectively.The control groups started at 18.51%,and progressed to 24.85% ,37.24% and 45.84% over the same interval.Conclusion STR was associated with improved LMF scores of stroke pa- tients.