Objective To investigate the aerosolized livability,aerosolized quantity and aerosol particle diameter,etc. of phage D29. Methods Phage D29 aerosol was produced in the aerosol test cabinet in negative pressure laboratory. TSI3321 aerodynamic particle sizer spectrometer was used for detecting the diameter of the aerosol particle. Bioaerosol particle median diameter was tested with Anderson six grade microorganism sampler. The aerosolized livability and quantity of phage D29 were determined by the variations of the concentrations and quantities of phage D29 before and after aerosolization.Results The aerodynamic diameter of phage D29 aerosol was 0.872 μm. Bioaerosol particle median diameter was 2.21 μm. The livability after aerosolization for 5,15,30,45,and 60 min were 89.78％,77.19％,48.86％,33.99％,30.12％,respectively. The aerosolized quantity of phage D29 was 232 μl/min. Conclusion The characters of the aerosolized livability,aerosolized quantity and aerosol particle diameter of phage D29 were thought to be suitable for further research into aerosol inhalation treatment of Mycobacterium tuberculosis infection in animals.

Objective To assess the value of functional magnetic resonance urography (fMRU)for the unilateral renal function in children with hydronephrosis.Methods Fourteen children with congenital hydronephrosis (unilateral hydronephrosis in 1 2 cases,bilateral hydronephrosis in 2 cases)examined by fMRU in Beijing Children′s Hospital,Capital Medical University,were enrolled.In 7 patients of them,diuretic renal scintigraphy (DRS)was per-formed within 1 0 days before fMRU examination.The following parameters in fMRU,as renal parenchymal volume,volu-metric differential renal function (vDRF),Patlak,Patlak differential renal function (pDRF),index of glomerular filtra-tion rate (GFR)and differential renal function based on index of GFR (gDRF),were calculated and analyzed.Statisti-cal analysis was performed by using SPSS 1 3.0.Results In 7 cases whose fMRU and DRS were examined,the indexes of GFR obtained from fMRU and GFR from DRS were well correlated (r =0.892,P <0.001 )in 1 4 kidneys.The gDRF determined by 2 methods on the left kidneys[the average was(46.80 ±1 9.20)% and(45.1 8 ±20.29)%,respective-ly]had no significant difference (t =0.051 6,P =0.624),which was also highly correlated (r =0.91 2,P =0.004). In 1 2 cases with unilateral hydronephrosis,vDRF,pDRF,index of GFR and gDRF in hydronephrotic side[(43.54 ± 9.61 )%,(42.80 ±1 0.83)%,(38.56 ±29.23)mL/min,(38.37 ±1 3.61 )%]were all less than those in the con-tralateral side[(56.46 ±9.61 )%,(57.1 9 ±1 0.83)%,(57.02 ±26.22)mL/min,(61 .63 ±1 3.61 )%](t =2.326, 2.300,2.422,2.960;P =0.040,0.042,0.034,0.01 3).However,there was no statistical difference in both renal pa-renchymal volume and Patlak between the hydronephrotic and the contralateral side kidneys(t =1 .765,1 .450;P =0.1 05,0.1 75).Conclusions fMRU is a very valuable examination method in evaluating single kidney function in children with congenital hydronephrosis,and able to demonstrate that gDRF,indexes of GFR,vDRF and pDRF decrease in the hydronephrotic kidney.

Objective To develop a micro-circumstance airtight cabin for in the study of biological aerosols detection with such functions as airflow control and temperature and humidity detection .Methods Wind speed sensors , temperature and humidity sensors , electrical control valves , high efficiency filters and the vacuum pump formed the micro-circumstance regulating system .The techniques of airflow direction control , temperature compensation , air pressure control and aerosol uniformity distribution were used .Numerical simulation of aerosol concentration distribution in the airtight cabin was achieved using Fluent software .The bioaerosol concentration in different locations was tested by experiments .Results The micro-circumstance airtight cabin consisted of an airtight cabin and a control cabin .The control cabin used a single-chip microprocessor to provide air supply and exhaust air to the airtight cabin in a seaparate exhaust mode and cyclic ventilation mode.It worked under a negative pressure condition .Through numerical simulation,the aerosols were distributed through-out the cabin after five minutes of generation and the bottom airflow arrived at the top .The distribution of aerosol concentra-tion was approximately uniform .Conclusion The micro-circumstance airtight cabin is suited to various bioaerosols testing research thanks to its negative pressure working without bioaerosol leakage .

Objective To evaluate the role of C-Jun N-terminal kinase (JNK) signal transduction pathway in spinal neurotoxicity induced by lidocaine in rats.Methods Seventy-two adult male Sprague-Dawley rats,weighing 220-260 g,were randomly divided into 6 groups (n =12 each):control group (group Ⅰ),sham operation group (group Ⅱ),JNK inhibitor group (group Ⅲ),dimethyl sulfoxide (DMSO) group (group Ⅳ),lidocaine group (group Ⅴ),and JNK inhibitor and lidocaine group (group Ⅵ).Group Ⅰ received no treatment.Intrathecal catheter was placed in the subarachnoid space in group Ⅱ.SP600125 25 μg and DMSO 20 μl were injected intrathecally in Ⅲ and Ⅳ groups,respectively.In group Ⅴ,10％ lidocaine 20 μl was intrathecally injected.SP600125 25 μg was injected intrathecally and 30 min later 10％ lidocaine 20 μl was injected intrathecally in group Ⅵ.Paw withdrawal threshold to yon Frey filament stimulation (PWT) and paw withdrawal latency to nociceptive thermal stimulation (PWL) were measured before intrathecal catheter was implanted (T0),before intrathecal administration (T1) and at 4,8 and 12 h and on 1,2,3,4,5 and 6 days after intrathecal administration (T2-10).At 24 h after intrathecal administration,4 rats were randomly chosen from each group and sacrificed.Their lumbar enlargements were removed for determination of phosphorylated JNK (p-JNK) expression (using Western blot) and neuronal apoptosis (by TUNEL).The apoptotic index was calculated.Results Compared with group Ⅰ,no significant difference was found in MWT and TWL in Ⅱ,Ⅲ groups and expression of p-JNK in Ⅱ and Ⅳ groups (P ＞ 0.05),MWT at T2-4,6-8 and TWL at T2-4,7 in group Ⅴ and MWT at T2-6 and TWL at T2-5 in group Ⅵ were significantly increased,the expression of p-JNK was down-regulated and the apoptotic index was decreased in group Ⅲ (P ＜ 0.05),and the expression of p-JNK was up-regulated and the apoptotic index was increased in Ⅴ and Ⅵ groups (P ＜ 0.05).Compared with group Ⅴ,MWT and TWL were significantly decreased,the expression of pJNK was down-regulated and the apoptotic index was decreased in group Ⅵ (P ＜ 0.05).Conclusion Activation of JNK signal transduction pathway is involved in spinal neurotoxicity induced by lidocaine in rats possibly through promoting neuronal apoptosis in the spinal cord.

Objective To investigate the immunogenicity of mycobacteriophage D29 (phage D 29) in guinea pig models with different delivery routes,and provide information for the application of phages in tuberculosis (TB) therapy.Methods Hartley guinea pigs were administrated with phage D29 through inhalation,intranasal drop or subcutaneous injection for 6 times within 35 days.7H9 broth aerosol inhalation and 0.85 ％ NaCl solution aerosol inhalation were set as solvent and negative controls,respectively.Anti-phage D29 neutralizing antibodies in sera collected weekly were measured by phage reduction neutralizing test (PRNT) and cytokine levels (interleukin-2,interleukin-4 and interferon-γ) were detected at day 35 by enzyme linked immunosorbent assay (ELISA).The data were analyzed by ANOVA and nonparametric test.ResultsNeutralizing antibodies were both negative in two control groups,while low-titer neutralizing antibodies (below 1 ∶ 100) appeared in inhalation and intranasal drop groups only at day 7 and day 14. Nevertheless, neutralizing antibodies were continuously detected in subcutaneous injection group,which increased rapidly and reached 1∶ 16 365.6 at day 35. After 35 days of experiments,serum concentrations of interleukin-2 (x2 =2.7605,P＞0.05),interleukin-4 (F=2.17,P＞0.05) and interferon-γ(F=0.75,P＞0.05) among three treatment groups and two control groups were all not significantly different.ConclusionsThe titer of anti-phage 29 neutralizing antibodies induced by inhalation or intranasal drop administration of phage D29 are both significantly lower than subcutaneous injection.Phage D29 administration doesn’t change the levels of cytokines,which indicates that it may not break the helper T cell (Th)1/Th2 balance.

OBJECTIVETo explore the effect of arteriosclerosis obliterans (ASO) blood stasis syndrome (BSS) serum on vascular endothelial cell injury and to study the regulation of Taohong Siwu Decoction (TSD) on it.

METHODSUmbilical vein endothelial cell culture system was established. The serum endothelial cell injury model with ASO BSS was prepared. Low, medium, and high concentrations TSD containing serums were respectively added. The endothelial cell proliferation activity was observed by MTT method. Ultrastructures of endothelial cells were observed under transmission electron microscope. Changes of intracellular calcium ion concentration and the cytoskeleton were observed under laser confocal microscope. Contents of ET, NO, and transforming growth factor beta1 (TGF-beta1) in endothelial cell culture supernatant were detected by ELISA.

RESULTSIn ASO BSS serum group endothelial cell proliferation activities decreased, the cell structure was obviously destroyed, calcium ion concentration increased, contents of ET, NO and TGF-beta1 increased significantly (P < 0.01), and ET/NO ratio was imbalanced. After incubating with TSD drug containing serum, endothelial cell proliferation activities and injured cell structures were obviously improved; ET, NO and TGF-beta1 levels decreased (P < 0.05, P < 0.01), ET/NO ratios approximated to the normal level.

CONCLUSIONThe main mechanism of TSD for treating ASO ASS lied in improving injured vascular endothelial cells and endocrine disorder.

Arteriosclerosis Obliterans ; Cell Proliferation ; Drugs, Chinese Herbal ; therapeutic use ; Endothelial Cells ; Humans ; Medicine, Chinese Traditional ; Serum ; Transforming Growth Factor beta1 ; metabolism ; Umbilical Veins

The aim of the manuscript was to optimize formulations and preparation technologies of cataplasm of white mustard seed varnish, and to evaluate its anti-asthma effect on rats. The single factor experiments included spreading thickness, types of crosslinking agents, dihydroxyaluminum aminoacetate amount, sodium polyacrylate amount, types of adhesive agents with human sense as the evaluation index. Blank cataplasm matrix was optimized by the orthogonal experiment with the amount of glycerine, citric acid, and sodium carboxymethylcellulose as the major influential factors. Initial adhesive force, peeling strength and human sense were as the evaluation index. The optimized formulation of blank cataplasm were as followings: glycerine-water-ethanol-PEG400-dihydroxyaluminum aminoacetate-citric acid-sodium carboxymethylcellulose-sodium carboxymethylcellulose 2 : 8 : 0.8 : 0.4 : 0.07: 0.15 : 0.1 : 0.5. The active ingredients of white mustard seed, corydalis, and gansui root were extracted by alcohol extraction method. Asiasarum volatile oil was extracted by oil extractor. The optimized drug loading amount was 11% with initial adhesive force, peeling strength and human sense as the evaluation index. Asthma rats model were established by sensitized with ovalbumin and nose-scratching time as the evaluation index. High dose (17%) group of drug-loaded cataplasm had the obvious inhibition effect on nose-scratching time of rats (P = 0.037 < 0.05). In comparison, middle dose (11%), low dose (4%) and positive-control groups had no obvious inhibitive effect on rats. White mustard seed cataplasm supplied a novel choice for anti-asthma therapy. And the overall pharmacodynamics assessment will be carried out on molecular level in near future.
Animals ; Anti-Asthmatic Agents ; administration & dosage ; chemistry ; Asthma ; drug therapy ; Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; Humans ; Male ; Mustard Plant ; chemistry ; Rats ; Rats, Sprague-Dawley ; Seeds ; chemistry

BACKGROUNDThe study was designed to investigate the status of molecular epidemiology of HCMV in Urumqi through genetic comparison of clinical isolates.

METHODSDNA sequences of 2.0-2.6 kb were amplified by polymerase chain reaction from three relatively conservative gene regions (DNA polymerase, glycoproteins H, and major immediate-early antigen) of 28 clinical HCMV strains and then were analysed by restriction enzymes.

RESULTSThe restriction patterns of the clinical isolates which did not have relation in epidemiology were greatly different, but the patterns of the clinical isolates related in epidemiology such as strains paired in mother and infant were quite similar. Of eight mother and infant pairs, from whom HCMV were isolated, four pairs showed identity of restriction profiles within each pair for all three amplified regions, four pairs showed differences between mother and infant.

CONCLUSIONThese results confirm the high degree of genetic variability among cytomegalovirus strains in Urumqi. Analysis of PCR-RFLP can indicate transmission of HCMV infection and facilitate its molecular epidemiologic studies.

China ; epidemiology ; Cytomegalovirus ; genetics ; isolation & purification ; Cytomegalovirus Infections ; epidemiology ; virology ; DNA-Directed DNA Polymerase ; genetics ; Humans ; Immediate-Early Proteins ; genetics ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Viral Envelope Proteins ; genetics

OBJECTIVETo discuss the effect of Taohong Siwu Decoction (TSD) in regulating functions of endothelial cells and treating arteriosclerosis obliterans (ASO).

METHODSThe ASO model was prepared by using high-fat diet plus intimal injury. They were randomly divided into the model group (n = 10), the normal control group (n = 9), the low dose TSD group (group A, n = 12), the middle dose TSD group (group B, n = 10), and the high dose TSD group (group C, n = 9). Eight weeks after modeling, the limb blood perfusion was observed using laser Doppler flowmetry. The arterial morphology was observed using light microscope and transmission electron microscope. The number of circulating endothelial cells (CECs) was determined using Percoll density gradient centrifugation method. Serum levels of TNF-alpha, IL-1, ET-1, and NO were detected using double antibody sandwich assay of enzyme linked immunosorbent assay (ELISA).

RESULTSThe ASO rat model was successfully established. Blood lipids levels significantly increased, the blood perfusion of left hind limbs significantly decreased, the number of CECs in the peripheral blood significantly increased, the arterial lumen was irregularly narrowed, the ultra-structure of vessel walls was damaged, serum levels of TNF-alpha, IL-1, and ET-1 significantly increased, and the serum level of NO significantly decreased in the model group, showing statistical difference when compared with the normal control group (P < 0.01). Compared with the model group, significant improvement in the aforesaid indices was shown in group B and C (P < 0.05, P < 0.01).

CONCLUSIONSThe injury and abnormal functions of endothelial cells is an important pathological process of ASO. As an effective recipe for treating ASO, TSD could protect vascular endothelial cells and improve the secretion function of vascular endothelial cells.

Animals ; Arteriosclerosis Obliterans ; blood ; drug therapy ; Diet, High-Fat ; adverse effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Endothelial Cells ; metabolism ; Endothelin-1 ; blood ; Endothelium, Vascular ; cytology ; Interleukin-1 ; blood ; Male ; Nitric Oxide ; blood ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; blood

Objective To establish a HPLC-MS/MS method for the determination of melatonin in human saliva.Methods Sabra samples and internal standard(itopride)were extracted by dichloromethane after alkalization.The analysis was conducted using a Sunfire C18 column with gradient elution and the column temperature iS 35 ℃.The mobile phase consisted of methanol and water with 0.1%formic acid at flow rate of 0.3 mL·min~(-1).Analysis in mass Was operated by the electrospray ionization.Results LLOQ of the method is 5.48 pg·mL~(-1)(S/N ＞10).The standard curve is linear in the range of 5.48～250.00 pg·mL~(-1)(γ2=0.9994).LLOD is 2 pg·mL~(-1)(S/N＞3).The RSDs of interday and intra-day are all less than 15%.The extraction recovery is closed to 60%.Conclusion The method is fast,precise and sensitive,and suitable for the determination of melatonin in human saliva.