Objective:To quantify the tryptase and T cell immunoglobulin mucin domain-3(TIM-3)double positive mast cells in hu-man chronic periodontitis tissue using double immunofluorescence staining.Methods:25 healthy controls,28 chronic mild periodontitis and 30 chronic advanced periodontitis patients were included.The gingival specimens were stained with HE for histology,and with double immunofluorescence staining for the identification of tryptase and TIM-3 double positive mast cells in gingival tissue.Results:In chronic periodontitis tissue the degree of gingival inflammation was significantly increased,the densities(cells/mm2 )of tryptase and TIM-3 double positive mast cells were significantly increased(P<0.05),in addition,that in chronic advanced periodontitis group was significantly higher than in the chronic mild periodontitis group(P<0.05).Conclusion:Tryptase and TIM-3 double positive mast cells has the similar tendency as the severity of periodontitis inflammation in human periodontitis tissue.Tryptase and TIM-3 double positive mast cells may play an important role in human chronic periodontitis.

Objective:To optimize the formula of metronidazole and borneol sustained-release films. Methods: The mass ratio of Rhizoma Bletillae polysaccharide and polyvinyl alcohol 124(PVA-124) and the dosage of glycerol and sodium carboxymethyl cellulose ( CMC-Na) as the influencing factors ,the appearance of films and the release rate of metronidazole at 0. 5h and 4h as the comprehen-sive evaluation indices, the formula of the film-forming materials was optimized by orthogonal test. Results:The mass ratio of Rhizoma Bletillae polysaccharide and PVA-124 was the key factor affecting the quality of the films. When the mass ratio of Rhizoma Bletillae pol-ysaccharide and PVA-124 was 20∶80, and the dosage of glycerol was 3. 5g and that of CMC-Na was 0. 14g, the appearance of the films was satisfied, the release rate of metronidazole at 0. 5h and 4h was 25. 17% and 69. 64%, respectively, and the average cumulative re-lease rate at 8h was over 90%. Conclusion:The films prepared by the optimized formula are flat, smooth, clean and moderately soft, which can not only remain the drug release characteristics of the films made by the original formula, but also exhibit the pharmacologi-cal effect of Rhizoma Bletillae.

OBJECTIVE:To investigate the preparation technic and method of quality control of Xinyoukexing tincture(XYKX) METHODS:To determine the content of Valaciclovir by spectrophotography,and podophyllotoxin by dual-wavelength spectrophotography RESULTS:In the range of 14 88～34 72?g/ml,the Valaciclovir concentration in XYKX was in direct proportion to the absorption at 310 4nm,C=44 5 905A+1 7 556,r=0 9 998 In the range of 17 50～50 12?g/ml,the podophyllotoxin concentration in XYKX was in direct proportion to the absorption at 292nm,C=97 7 981Ap292+1 3 614,r=0 9 999,AP292=As292-As325 CONCLUSION:The preparation technic of XYKX is simple and the method of quality control is feasible

The genome of CK/CH/SD09/005, an isolate of infectious bronchitis virus (IBV), was characterized to enable the further understanding of the epidemiology and evolution of IBV in China. Twenty-five pairs of primers were designed to amplify the full-length genome of CK/CH/SD09/005. The nucleotide sequence of CK/CH/SD09/005 was compared with reference IBV strains retrieved from GenBank. The phylogenic relationship between CK/CH/SD09/005 and the reference strains was analyzed based on S1 gene sequences. The complete genome of CK/CH/SD09/005 consisted of 27691 nucleotides (nt), excluding the 5' cap and 3' poly A tail. The whole-genome of CK/CH/SD09/005 shared 97 - 99% nucleotide sequence homology with the GX-NN09032 strain, which was the only complete genome that was closely related to CK/CH/SD09/005. When compared with all reference strains except GX-NN09032, CK/CH/SD09/005 showed the highest similarity to ck/CH/LDL/091022 and SDIB821/2012 (QX-like) in the replicase gene (Gene 1) and 3'UTR, with a sequence identity rate of 97% and 98%, respectively. However, CK/CH/SD09/005 exhibited lower levels of similarity with ck/CH/LDL/091022 and SDIB821/2012 in S-3a-3b-3c/ E-M-5a-5b-N with a sequence identity of 72% - 90%. CK/CH/SD09/005 showed the highest level of nucleotide identity with Korean strain 1011, and Chinese strains CK/CH/LXJ/02I, DK/CH/HN/ZZ2004 and YX10, in ORF 3c/E (97%), 5a (96%), 5b (99%) and N (96%), respectively. ORFs 3a, 3b and M of CK/CH/SD09/005 exhibited no more than 90% homology with the reference strains, excluding GX-NN09032. The phylogenic analysis based on the S1 gene revealed that CK/CH/SD09/005 and 39 published strains were classified into seven clades (genotypes). CK/CH/SD09/005 was distributed in clade IV with several isolates collected between 2007 and 2012. CK/CH/SD09/005 showed 66% - 69% and 72% - 81% nucleotide identities with the IBV strains of other six clades in the S1 and S2 subunits, respectively. More over, multiple substitutions were found throughout the entire S gene of CK/CH/SD09/005, while insertions and deletions were located within the S1 gene. These results indicated that CK/CH/SD09/005 is a novel variant that may be derived from the QX-like strains that are prevalent in China. Multiple genetic mechanisms, including recombinations, mutations, insertions and deletions, are likely to have contributed to the emergence of this IBV strain.
Animals ; Chickens ; China ; Coronavirus Infections ; veterinary ; virology ; Genome, Viral ; Genomics ; Infectious bronchitis virus ; classification ; genetics ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Poultry Diseases ; virology ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics

Objective To optimize several key factors in preparing protein microarray,and establish microarrays to detect autoantibodies.Methods Four factors,including incubation time of the microarray after spotting,the blocking reagent,the length of blocking time,the length of time conjugating with the second-an- tibody,were selected to carry out orthogonal optimization,then we determined the optimal spotting concentra- tion of antigen and the best dilution of serum with the optimized conditions obtained from the above.On the basis of the optimized conditions,we prepared microarray to detect six autoantibodies simultaneously,which were compared with IIF and ELISA.Results The best conditions we determined from the experiment were: incubating the spot microarray for 120 min,PBST containing 1% BSA and 2.5% saccharose as the blocking reagent and blocking for 30 min,reacting with the second-antibody for 30 min,the appropriate spotting con- centrations of six antigens differed from 100?g/ml to 300?g/ml and the best serum dilution was 1:2.Com- pared with IIF for ANA detection,the sensitivity of the microarray was 88.6%,the specificity was 83.3%,and compared with ELISA in detecting the other five autoantibodies,the sensitivity of the microarray was between 80.0% and 88.2%,and the specificity was between 90.2% and 98.6%.Conclusion The optimization condi- tions we obtained are suitable for preparing protein microarray,and the detection sensitivity and specificity of autoantibodies by microarray are consistent with the conventional methods in general.The microarray for de- tecting autoantibodies is applicable in the clinical diagnosis of autoimmune disease.

Hamartoma ; complications ; surgery ; Humans ; Hydrocephalus ; etiology ; Hypothalamic Diseases ; complications ; surgery ; Infant ; Male ; Postoperative Complications ; etiology ; Puberty, Precocious ; etiology ; Subdural Effusion ; etiology

BACKGROUND:Vascular endothelial growth factor is a potent angiogenesis and permeability inducible factor. Vascular endothelial growth factor 165 and vascular endothelial growth factor 121 are mainly expressed in vivo, with a strong role of angiogenesis.
OBJECTIVE:To observe the feasibility of vascular endothelial growth factor 165 gene transfected bone marrow mesenchymal stem cel s to differentiate into vascular endothelial cel s.
METHODS:Bone marrow derived mesenchymal stem cel s were isolated and col ected from 50 g Sprague-Dawley rats,and identified by flow cytometry. The plasmid pGLV-EF1a carrying a vascular endothelial growth factor 165 gene was transfected to the mesenchymal stem cel s using lentiviral. Expression of green fluorescent protein was observed under a fluorescence microscope.
RESULTS AND CONCLUSION:After 12 hours of transfection, expression of green fluorescent protein was observed, increased at 48 hours, peaked at 72 hours and gradual y declined thereafter. Results prove that vascular endothelial growth factor 165 gene transfected bone marrow mesenchymal stem cel s have the expression of green fluorescent protein, indicating successful transfection. It is feasible to induce bone marrow mesenchymal stem cel s to differentiate into vascular endothelial cel s.

ObjecfiveTo investigate clinical features and imaging manifestation in patients with posterior reversible eneephalopathy syndrome (PRES) to improve its recognition.MethodsSix patients with PRES were enrolled,four women with history of end-stage renal disease,kidney transplantation,eclampsia,or systemic lupus erythematosus (SLE) and two men with history of chemotherapy or hypertension.All of them underwent multi-serial MR imaging (T1 WI,T2 WI,FLAIR) and post-contrast T1 WI.Three cases also underwent CT scan and gadolinium enhancement. ResultsAll the six cases of PRES had different inducing causes such as acute hypertension,preeclampsia or eclampsia, taking immunosuppressive agents or steroids.and their clinical symptoms were characterized by sudden occurrence of headache,eclampsia or seizure of epilepsy,altered melltal status,visual disturbances.Clinical symptoms were died out in about one week after prompt and appropriate treatments for high blood pressure.or removal of precipitating factors,or treatment for epileptic seizures or status epilepticus.MRI and CT scanning demonstrated multifocal subcortical white lesions in bilateral parieto-occipital lobes (six cases), bilateralfrontal lobes (two cases),bilateral post temporal lobes (two cases) and left cerebellum (one cases).and cortical involvement (two cases).All lesions appeared unenhanced with gadolinium enhancement. FoHow-up by MRI showed decreased abnormal signs and small infarct foci were left in the cortex-subeortex of one case.ConclusionsPRES is a clinico-neuroradiologieal transient condition, usually benign and reversible in nature.Completely clinical and radiographic recovery Can be achieved with prompt antihypertensive treatment or removal of precipitating factors and supportive care,but delayed diagnosis and therapy Can result in cerebral infarct with neurological sequelae.

OBJECTIVETo observe the effect of Tetramethyl pyrazine (TMP) on the cytokines and inflammatory mediators in the serum and the synovial fluid of collagen-induced arthritis (CIA)rats, and further to investigate its possible mechanisms for treating rheumatoid arthritis (RA).

METHODSType II CIA rat model was established. Rats in the TMP group were administered with TMP at 50 mg/kg and 100 mg/kg, once daily. Dexamethasone at 2.0 mg/kg was intramuscularly injected to those in the Dexamethasone treated group, once daily. Normal saline at 2 mL/kg was given to those in the normal control group and the model group, once daily. All medication was started from the 7th day, lasting to the 35th day. CIA rats' foot swelling degree was observed. Contents of serum IL-1, IL-6, IL-2, NO and PGE2in the synovial fluid were detected by radioimmunoassay and nitrate reduction method.

RESULTSCompared with the normal group, the foot swelling obviously increased, contents of NO and PGE2 in the synovial fluid were obviously elevated in the model group (P < 0.01). Compared with the model group, the foot swelling could be obviously inhibited by 100 mg/kg TMP and Dexamethasone; serum levels of IL-1 and IL-6 obviously decreased, serum IL-2 level obviously increased, contents of NO and PGE, decreased (P < 0.01). TMP 50 mg/kg could obviously inhibit the foot swelling of CIA rats (P < 0.01). There was no statistical difference in other indices (P > 0.05).

CONCLUSIONSTMP at 100 mg/kg showed obvious inhibition on CIA rats. Its inhibitory effect might be correlated to inhibiting activities of endogenous cytokines and the generation of inflammatory mediators in inflammation local regions, improving contents of anti-inflammation cytokines, and inducing the balance of the inflammatory cytokine network.

Animals ; Arthritis, Experimental ; blood ; metabolism ; Dinoprostone ; metabolism ; Female ; Interleukin-1beta ; blood ; Interleukin-2 ; blood ; Interleukin-6 ; blood ; Male ; Nitric Oxide ; metabolism ; Pyrazines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Synovial Fluid ; metabolism

Using the Gram-positive Staphylococcus aureus as the indictor bacterium,fourteen antibacterial strains were initially obtained by the bilayer-media screening method from the raw milk samples,and one isolate was found to exhibit the higher antibacterial activity against the indicator. This isolate was further studied on its individual and cultural morphology features,partial physiological and biochemical reaction activities,G+C content,the sequence features of the 16S rDNA and the species-specific N-acetylmuraminidase gene(acmA) ,consequently,it was identified as the Lactococcus lactis subsp. lactis strain,named as MB191. An evaluation of the antimicrobial spectra of MB191 was subsequently performed,it showed the remarkable activities against not only the tested Gram-positive bacteria,but also several Gram-negative bacteria including Pseudomonas syringae and P. fluorescens,as well as the yeast Debaryomyces hansenii,which was a distinctive feature that was not reported prior to this study.