Objective:To study performed of develop a mice model of allergic diseases induced by crude extractings from populus pollen.Methods: A total of 60 BALB/c mice were divided randomly into there groups:normal control group,Albumin Egg(OVA) group and populus pollen model group with 20 in each.Mices were repeatedly sensitized by intraperitoneal injections of OVA or crude populus pollen extract every 5 d for four doses.Five days after the last sensitization,mices were repeatedly challenged by once daily antigen from 21-25 d.The changes of inflammatory cells in bronchoalveolar lavage fluid(BALF) were stained with hematoxylin and eosin(HE) to evaluate the degree of allergic inflammation.PAS staining was used to observe the secretion of airway mucus;The changes of the nasal mucosas and lungs of mice were stained with HE to evaluate the degree of allergic inflammation.And the average optical density of IL-4 and IFN-γ positive cells in lung tissue was measured by immunohistochemistry.The total IgE in the serum was also measured by enzyme-linked immunoassay(ELISA).Results: Compared with the mice in normal control group,those in OVA group and model group developed obvious allergic inflammation in the nasal mucosas and lungs,and increased airway mucus secretion.The number of inflammatory cells including eosinophil and neutrophils markedly increased in BALF smear.The average optical density of IL-4 positive cells in lung tissue was all increased in OVA group and model group compared with those in normal control group,and the average optical density of IFN-γ in lung tissue was on the contrary.The total IgE in the serum were all increased in OVA group and model group compared with those in normal control group,and the IFN-γ in the serum was significantly reduced in OVA group and model group compared with those in normal control group.Conclusion: Taken together,crude populus pollen extract can successfully induce a mice model of allergic diseases.This model is a useful tool in studying the mechanisms of allergic disease.

Objective To investigate the effect of periurethral injection of insulin-like growth factor(IGF)-Ⅰ on the expression of IGF-Ⅰ and IGF-Ⅱ mRNA during regeneration period following urethral sphincter muscle injury in female rats. Methods Model of urethral sphincter muscle injury was made in female virgin SD rats (n=50) by intravaginal balloon inflation. Then the rats were divid-ed randomly into treatment group (n= 25) and control group (n= 25), treatment group accepted peri-urethral injection of 1.0μg human IGF-Ⅱ to the middle urethral muscle, control group accepted nor-mal saline injection. Five rats in each group were sacrificed at 2, 4, 6, 8, 14 day respectively and the whole urethra specimens were processed for RT-PCR to detect the expression of IGF-Ⅰ ,Ⅱ mRNA. A normal control group (n = 5) was set without intravaginal balloon inflation and injection. Results The expression of IGF-Ⅰ mRNA in control group increased at day 4, 6, 8, 14, the IGF-Ⅰ/β-actin ra-tios were 0. 58±0.15, 1.73±0.31, 2.30±0.29, 0. 46±0. 06. The expression of IGF-Ⅰ mRNA in treatment group increased at all time points, as 0. 69±0.21, 1.45±0.17, 2.25±0.45, 2.90±0.49, 1.92±0. 31. The difference was significant on day 4, 14(P<0.01), and day 8 (P<0.05) compared with the control group. The expression of IGF-Ⅱ mRNA in control group increased at day 4, 6, 8, as 0.42±0. 14, 1.51±0. 59, 1.31±1.04. The expression of IGF-Ⅱ mRNA in treatment group in-creased at day 4, 6, 8, 14, as 1.04±0.23, 1.94±0.29, 1.75±0.41, 0. 81±0.15. The significant difference was noted on day 4 (P<0. 01)compared with the control group. No expression of IGF-Ⅰand Ⅱ mRNA in the normal control group. Conclusions The expression of endogenous IGF-Ⅰ and IGF-Ⅱ mRNA was up-regulated by periurethral injection of IGF-Ⅰ during regeneration period follow-ing urethral sphincter muscle injury in female rat. Our findings suggest that IGF-Ⅰ facilitates the re-generation of the urethral muscles and may play a role in treatment of stress urinary incontinence in-duced by urethral sphincter muscle dysfunction.

A novel genomic island (GI) in Streptococcus suis serotype 2(SS2) was identified, which resided in the highly virulent strains but not in the hypo-virulent strains or avirulent strains of SS2 of the Chinese isolates. This newly discovered GI strain was designated as SSGI4 and its whole length of genome was 11 269 bps, sharing the typical properties of pathogenicity islands, such as the distinct G+C content, a mosaic architecture characteristics and the specificity for virulent isolates. There were 11 genes within SSGI4, in which some genes were putative cell surface protein genes and others were amino acid-binding protein genes. Our finding sheds light on the investigation of horizontal gene transfer in SS2 and their influence on pathogenicity.

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 explore the predicted performance of cTnI for outcome or severity in children with sepsis.Methods 374cases of children with sepsis were collected in pediatric intensive care unit (PICU) in our hospital from August 2012to June 2015.The patients were dividided into the common sepsis group, severe sepsis group and sepsis shock group according to the sepsis severity, and improved group, uncured group and death group according to outcome, and the cTnI>0.01μg/mL group and the cTnI≤0.01μg/mL group according to the levels of cTnI.Data on cTnI, PCT, CRP, Cr, Lac, PaO2/FiO2, BUN, PT, INR, WBC and PLT were collected in this study.Results The level of cTnI was significantly higher in children with septic shock (P<0.05) .The level of cTnI in improved group was significantly lower than those of uncured group and death group (P<0.05) .The incidence of severe sepsis and septic shock in the cTnI>0.01μg/mL group was significantly significantly higher than that of the cTnI≤0.01μg/mL group.The levels of Lac, PT and INR in the cTnI>0.01μg/mL group were significantly higher than that of the cTnI≤0.01μg/mL group (P<0.05) .A positive correlation between the level of cTnI and Lac (r=0.324) , or PT (r=0.291) , or INR (r=0.340) were found in the study (P<0.05) .Conclusion Sepsis is prone to be associated with myocardial injury, which is related to the severity and prognosis of sepsis.Insufficient circulatory perfusion, metabolic imbalance and abnormal coagulation function may be the reasons for the rise of cTnI and myocardial injury in children with sepsis.

Objective: To analyze the clinical characteristics of hepatic flare and evaluate efficacy of antiviral treatment in pregnant women with chronic HBV infection. Methods: A single-center, open-label, prospective study was conducted, and pregnant women with chronic HBV infection were enrolled. Liver function, HBV serum markers and HBV DNA of pregnant women with chronic HBV infection were reviewed during every 4 to 12 weeks of gestation period. The proportion and clinical characteristics of hepatitis flare during pregnancy were observed. Logistic regression analysis was used to predict hepatic flare in pregnant women with chronic HBV infection. Antiviral therapy with telbivudine (LdT) or tenofovir dipivoxil (TDF) was used to treat hepatic flare during pregnancy. Sequential entecavir (ETV) or TDF was applied after the delivery. Treatment course and drug withdrawal in pregnant women with hepatic flare was the same as those of the general patients with chronic hepatitis B. Liver function, HBV serum markers and HBV DNA were measured in pregnant women with hepatic flare at different time points (4, 12, 24 and 52 weeks). A t-test was used to compare the hepatic flare in pregnant women with and without hepatitis group. HBsAg and HBeAg were used to quantify the receiver operating characteristic (ROC) curve of pregnant women with hepatic flare during pregnancy. Area under the ROC curve was used to calculate the optimal cut-off value corresponding to the maximum sensitivity and specificity of the ROC curve. Results: Of the 220 pregnant women with chronic HBV infection, 55 (25%) had hepatitis flare during pregnancy and received antiviral treatment. Among the 55 women with hepatic flare during gestation, 47 (85.46%) had hepatic flare in the mid-second trimester (12-24 weeks); average peak value of alanine aminotransferase (ALT) was 220.62 U/L, and the average peak value of ALT in 32 cases (58.18%) of pregnant women with hepatic flare was between 2–5 × ULN. HBsAg and HBeAg quantification were significantly lower in pregnant women with hepatic flare during pregnancy than with non-hepatitis (t = -3.745, P < 0.001; t = -2.186, P = 0.030). Multivariate logistic regression analysis showed that pregnant women with HBeAg < 3.065 log10 s/co were 7.576 times more likely to have hepatic flare during pregnancy (95% confidence interval: 3.779-15.190). ALT normalization, undetectable HBV DNA levels, HBeAg loss and HBeAg seroconversion in 55 pregnant women with hepatic flare at 52-week treatment was 100% (55/55), 74.55% (41/55), 47.27% (26/55) and 41.82% (23/55), respectively. HBsAg quantification at 52 weeks was significantly lower than baseline HBsAg quantification (3.32 + 0.37) log₁₀ IU/ml and (3.95 + 0.40) log₁₀ IU/ml; t = 8.465, P < 0.001). Conclusion Hepatic flare often occurs in the second trimester of pregnancy in pregnant women with chronic HBV infection and baseline HBeAg quantification is an independent predictor of hepatic flare. HBeAg seroconversion rate increased at 52 weeks after antiviral therapy.

OBJECTIVE: To investigate the advantages of nitric acid digestion method and its differences with the traditional method. METHODS: Ethanol was used to fully fix the testing sample. About 80-100 g of the testing samples were cut into pieces and digested with nitric acid. It was then centrifuged and washed to remove organic components. Smears were prepared and examined under the light microscope. RESULTS: The diatom had been identified with clear striations, counted conveniently and classified easily. CONCLUSION The improved nitric acid digestion method is not only simple with a higher successful rate of detection, but also can prevent interference from contamination. It can improve the stability of the experimental results, avoid harm to human and environment, and provide higher safety in the course of experiment.
Autopsy ; Diatoms/isolation & purification* ; Drowning ; Forensic Pathology ; Humans ; Kidney/metabolism* ; Liver/metabolism* ; Lung/metabolism* ; Nitric Acid/chemistry* ; Postmortem Changes ; Tissue Fixation/methods*

Objective To evaluate the effects of polishing on surface roughness (Ra) and surfacefree energy (SFE) of three direct tooth-colored restorative materials.Methods Three direct tooth-colored restorative materials,composite resin(CR) ( Filtek Z350,3M ESPE,America),pre-reacted glass ionomer (Giomer) ( Beautifil Ⅱ,Shofu,Japan) and resin-modified glass ionomer ( RMGIC ) ( Fuji Ⅱ LC,GC,Japan),were placed in cuboidal models (8 mm × 8 mm × 2 mm),covered with mylar film and light-cured.The samples ( n =14) were polished with coarse,medium,fine and superfine instruments in series ( grit 60 μm→grit 30 μm→grit 20 μm→grit 7 μm).Surface roughness (Ra,μm) was determined with a profilometer.Surface contact angles (CA) with Glycerol,Ethylene glycol and distilled water were measured using an image acquisition system and surface-free energy was calculated.Ra and SFE of different materials before and after polishing were compared by SPSS one-way ANOVA (LSD/Dunnett T3 ).Results Before polishing,Ra and SFE of CR [ (0.098 ±0.019) μm,(31.78 ±2.14) mN · m-1] and Giomer [ (0.093 ±0.020) μm,(32.63 ±2.12) mN · m-1] were both lower than those of RMGIC [ (0.134 ±0.019) μm,(40.22 ± 1.05 ) mN · m- 1 ].After polishing,all three materials showed increased Ra,and there were significant differences among three materials(P ＜0.05):CR[ (0.141 ±0.018) μm] ＜ Giomer [ (0.185 ±0.013) μm] ＜ RMGIC [ (0.494 ± 0.075) μm].However,all three materials showed decreased γSTOT after polishing.There was significantly different among the materials ( P ＜ 0.05 ):Giomer [ ( 26.60 ±5.61) mN·m-1] ＜ CR [(29.16±5.73) mN· m-1] ＜ RMGIC [(32.30±3.83) mN · m-1].Conclusions The surface roughness and surface free energy were material type and polishing technique dependent.

Objective To investigate early adhesion of Streptococcus mutans (Sm) (UA159) on the surface of direct tooth-colored restorative materials.Methods Three direct tooth-colored restorative materials,composite resin (CR,Filtek Z350,3M ESPE,America),pre-reacted glass ionomer (Giomer,Beautifil Ⅱ,Shofu,Japan) and resin-modified glass ionomer (RMGIC,Fuji Ⅱ LC,GC,Japan),were placed in fabricated models(8 mm × 8 mm ×2 mm),covered with mylar film and light-cured.The samples (n =5) were polished with disks in series (grit 60 μm、grit 30 μm、grit 20 μm、grit 7μm) (Supersnap,Shofu,Japan),then coated with artificial saliva for 2 h.Adhesion of Sm(UA159) on the surface of samples was investigated using confocal laser scanning microscopy(CLSM) in conjunction with fluorescence staining technique after 2 h.The area covered by dead and vital adhesive bacteria(Ad,Av) and dead/vital bacteria ratio(Ad/Av) of different materials were measured with Image-Pro Plus 5.1 and compared using SPSS (K-Independent-Samples Tests/Kruskal-Wallis H).Results Under CLSM,adhesive bacteria,especially vital ones,were observed on the surface of all the three materials.Compared with RMGIC,CR and Giomer showed less dead and vital bacterial adhesion (P ＜ 0.05):Ad [M (Q)]:Filtek Z350 [0.007％(0.037％)],Beautifil Ⅱ [0.011％ (0.028％)] ＜ FujiⅡ LC[0.082％(0.082％)].Av[M(Q)]:Filtek Z350 [0.103％ (0.162％)],Beautifil Ⅱ [0.067％ (0.181％)] ＜ Fuji Ⅱ LC [0.690％(0.482％)].However,Ad/Av[M(Q)] of Beautifil Ⅱ [0.250(0.188)] was lower than the other two,Filtek Z350[0.069(0.125)] and FujiⅡ LC[0.114(0.051)] (P＜0.05).Conclusions Early adhesion of Sm on the surface of direct tooth-colored restorative materials were material type dependent.

OBJECTIVETo evaluate the bioequivalence of tiopronin enteric capsules (testing preparation, T) versus tablets (reference preparation, R).

METHODSA single oral dose of tiopronin enteric capsules or tablets at 200 mg was administered in 2 groups of Chinese healthy volunteers (n=9) in a randomized crossover design at the interval of 2 weeks. The plasma concentrations of tiopronin were measured by HPLC-MS/MS, and the pharmacokinetic parameters were calculated by DAS 2.0 program. The bioequivalence between the two preparations was evaluated.

RESULTSThe main pharmacokinetic parameters were as follows: C(max)(microg.ml(-1)) 3.612-/+1.2393 (R), 3.644-/+1.540 (T); t(max) 4.333-/+1.0853 (R), 3.611-/+1.420 (T); t((1/2))(h) 18.245-/+11.270 (R), 23.403-/+10.500 (T); AUC0-t (microg.h.ml(-1)) 18.732-/+6.92318 (R), 18.713-/+6.585 (T); AUC0-infinity (microg.h.ml(-1)) 21.900-/+7.31220 (R), 20.780-/+7.965 (T). The relative bioavailability of tiopronin enteric capsule was 103.712-/+23.956%, with 90% confidential intervals of ln(AUC0-->72), ln(AUC0-infinity) and ln(C(max)) of 91.1%-111.8%, 96.8%-118.3%, and 85.1%-113.0%, respectively.

CONCLUSIONThe tiopronin enteric capsules were bioequivalent to the tablets.

Biological Availability ; Capsules ; Cross-Over Studies ; Health ; Humans ; Linear Models ; Male ; Reproducibility of Results ; Therapeutic Equivalency ; Tiopronin ; pharmacokinetics ; Young Adult