Objective To investigate the biochemical properties of rat improved perfusion-acellular pancreatic bioscaffold (APB).Methods The fresh pancreas from 10 rats were perfused through portal vein.The histological structure,ECM composition and DNA quantification of APB were evaluated.For the biocompatibility study,a 0.5 cm2 APB construct was surgically placed within a dorsal subcutaneous pocket of mice.Results The pancreatic tissue become translucent and the macroscopic three-dimensional architectures of native pancreas are retained after decellularization.The extracellular matrix (ECM) ultrastructures were well preserved.Immunofluorescence staining showed that collagen Ⅰ,Ⅳ,fibronectin and laminin were maintained in the APB after decellularization.DNA quantification of APB decreased significantly (P ＜ 0.05).The histological analysis of the subcutaneous implantation site showed the presence of immunological response surrounding the partially degraded APB.The histological remold score was 10.4 ± 1.8 at 14 days and 13.8 ± 1.3 at 28 days.The proliferation of AR42J cell line with the expression of amylase as the marker of exocrine acinar cells can be detected on the recellularized APB.Conclusion APB in this study met the stringent requirement to define a successful decellularization.The technique allowed the efficient generation of APB with preserved 3D architecture and represented a biocompatible scaffold capable of integrating within host tissue.

Near-infrared (NIR) was used as rapid analysis method to identify the sulfur fumigated Puerariae Lobatae Radix. NIR spectra of the cross-sectional and longitudinal selection of samples were acquired. Principal component analysis was conducted. The samples were randomly selected. The different pretreatment methods were compared. Discriminant models were established for every type of spectra to calculate the recognition rate. The orthogonal test and nonparametric test were used to test data normality. The result showed that absorbance values of different sections were different due to the different structure, and the raw spectra were analyzed by PCA method. The result founded that the cumulative contribution rate was arrived at 99.2% while the PC numbers were arrived at 3. The pretreatment method based on the MSC + 1D + Savitzky-Golay was the best to establish the model. For the 50 models constructed with cross-section and longitudinal spectra and total spectra, the recognition rate were (94.4 ± 0.66)%, (94.4 ± 0.66)%, (95.3 ± 0.65)%, respectively, and no difference was observed. The NIR method could be used to identify the sulfur fumigated Puerariae Lobatae Radix.
Chemistry, Pharmaceutical ; methods ; Discriminant Analysis ; Drugs, Chinese Herbal ; chemistry ; Fumigation ; Pueraria ; chemistry ; Spectroscopy, Near-Infrared ; Sulfur ; chemistry

Near-infrared (NIR) diffuse reflectance spectroscopy was used to analyze the impact of multi-class particle size of Chinese material medica (CMM) based on the spectral characteristics in overtone and combination region. Several types of CMM (60, 80, 100,120 mesh) were subjected to NIR spectra analysis. Spectral reproducibility was examined after sample repackage. The result showed that the effects of particle size on the NIR spectra were different according to different bands, in the combination region and first combination-overtone region. Spectroscopy intensity was proportional to the particle size and influence of particle size was greater as the wavelength increased. While in the second combination-overtone region, it was inversely proportional to particle size. To the sampling loading error, the result indicated that when the mesh number was larger than 60 mesh, the error was small. The appropriate particle size was clarified to guarantee the accuracy and reliability of NIR diffuse reflectance spectroscopy in CMM.
Drugs, Chinese Herbal ; analysis ; Materia Medica ; analysis ; Particle Size ; Spectroscopy, Near-Infrared ; methods

ObjectiveTo investigate the distribution and effect on antibiotic resistance of the novel cell wall anchored protein-encoding gene sasX.MethodsA total of 300 S.aureus isolates were randomly collected from inpatients with S.aureusinfection in ShanghaiHuashanhospitalin 2004, 2007and 2010.Meanwhile,170 S.aureus isolates from the nasal swabs of healthy people were collected as part of a population-based community prevalence study.Typing of S.aureus isolates were identified by using multilocus sequence typing (MLST) and S.aureus-specific staphylococcal protein A typing (spa typing).Determination of oxacillin MICs was used to screen MRSA.PCR and sequencing were used to analyze sasX gene.The effect on antibiotic resistance of sasX gene was detect by disc agar diffusion drug sensitive test.ResultsThe major clonal types of the 300 S.aureus isolates collected from inpatients with S.aureus infection were ST239 ( 110,36.7％) and ST5 (122,40.7％).From 2004 to 2010,the percentage of isolates from inpatients with S.aureus infection was increased from 17％ to 39％,but sasX was only found in 0.59％ of the S.aureus isolates from the nasal swabs of healthy people.The percentage of sasX positive was increased from 47.2％ to 83.8％ in ST239.The percentage of sasX positive MRSA was increased from 26.4％ to 50.8％,but the percentage of sasX positive MSSA was about 10％.Antibiotic resistance of sasX positive strains were higher than that of sasX negative strains.Conclusions SasX gene is mainly detected in nosocomial pathogenic S.aureus and it is a possible virulence factor of S.aureus in hosptal setting.The presence of sasX gene is related to antibiotic resistance.For better understanding the real function of this novel gene,further studies such as expression of the encoded protein should be carried out.

Objective To observe the incidence of fragmented QRS complex (fQRS)and ST Segment depression fQRS (STD fQRS)during the first 48 hours after non-ST elevation myocardial infarction(NSTE MI)and discuss the value of predicting mortality in patients with NSTE MI .Methods Based on the ECGs ,the patients with NSTE MI were divided into two groups :fQRS and non fQRS group .And then fQRS group was divided into two sub-groups :STD fQRS and non-STD fQRS group .Their mortality was studied during long-term follow-up .Results (1)731 patients with NSTE ACS [the NSTE MI group(n=609) and the UA group(n=122)] were studied .The incidence of fQRS in the NSTE MI group was higher than that of the UA group .(2)All cause mortality in the fQRS group were higher than that in the non-fQRS group ,and all-cause mortality in the STD fQRS group were higher than that in the non-STD fQRS group ,all the above results were not only in the early stages of NSTE MI ,but also in the long term fol-low-up .(3) Multivariate Cox regression analysis revealed that STD fQRS was an independent significant predictor for all cause mortality ,but not of the fQRS .Conclusion The STD fQRS may be an independent predictor of mortality in patients with NSTE MI .

Objective To investigate the clonal types of Staphylococcus aureus collected from 2004to 2010 in patients with blood stream infection from a Grade A tertiary care hospital in Shanghai as well as the dynamic changes and to detect the variation in antimicrobial resistance and virulence-gene content in different strain types.Methods A total of 103 nonduplicate S.aureus isolates were collected from 2004 to 2010 from inpatients with S.aureus blood stream infection from Shanghai Huashan hospital.Determination of oxacillin MICs and the type of SCCmec gene were used to screen MRSA.Typing of S.aureus isolates was identified by using multilocus sequence typing(MLST) and S.aureus-specific staphylococcal protein A typing(spa typing),PCR was used to detect the antimicrobial resistance and virulence-gene.Results Sixtysix isolates(64.1%) MRSA were detected in 103 nonduplicate S.aureus isolates,and 35 isolates were MRSA with SCCmec type Ⅱ ,Twenty-nine isolates were MRSA with SCCmec type Ⅱ,two isolates were MRSA with SCCmec type Ⅳ,Thirty-seven isolates(35.9%) were MSSA.Thirty-three MRSA isolates were ST5,Twenty-nine MRSA isolates were ST239,two MRSA isolates were ST59,one MRSA isolates was ST641 and one MRSA isolates was ST6.All of the other clones belonged to MSSA.The percentage of ST5 and ST239 were decreased significantly after 2009(ST5 was decreased from 52.9% to 15.4%; ST239 was decreased from 61.1% to 15.4%),and new clonal types MSSA increased significantly(in 2009,the percentage of ST7 was 41.7%; new clonal types such as ST188 and ST15 were detected in 2010).In 2010,it was shown that 84.6% of MSSA were isolated from S.aureus blood stream infection,nineteen isolates(18.4%) harbored mupA gene and 41 isolates(39.8%) harbored qacA/B gene in 103 nonduplicate S.aureus isolates.It was shown that 70.6% ST239 harbored qacA/B gene.Four isolates of ST398 and 1 isolates of ST9were detected which were originally from animal.There was no significant difference of the virulence gene presence in the same strain types except sasX、lukSF and arcA genes,but there were a lot of genes which were restricted to different genomic background.Conclusions The percentage of ST5 and ST239 were decreased and new clonal types of MSSA were increased significantly in S.aureus blood stream infection,antimicrobial resistance and virulence-gene were restricted to different clonal types.

Objective To determine whether the novel surface-anchored protein SasX promotes aggregation of S.aureus and adherence of S.aureus to human nasal epithelial cells.Methods MRSA ST-239 HS770 sasX gene mutant ( HS770 △sasX) and complement [ HS770 △sasX (pRBsasX) ] were gotten by gene knock-out and complement methods.The aggregation ability of S.aureus was observed through microscope.By adherence assay which was used for detection of the adherence ability of wild type and mutant to human nasal epithelial cells and blocking experiments which detected the ability of the purified recombinant SasX protein in blocking the adherence of S.aureus to human nasal epithelial cells,we investigated the influence SasX on colonization of S.aureus.Results Compared to wild type,HS770△sasX showed a reduction in cell aggregation,while the complement had no difference with wild type in aggregation. HS770 △sasX showed a significant reduction of adherence to human nasal epithelial cells compared to wild type ( P＜0.01 ),and the complement showed a very clear increasement of adherence to human nasal epithelial cells compared to wild type(P＜0.01 ).Preincubation of nasal epithelial cells with the purified recombinant SasX protein inhibited S.aureus binding significantly.Conclusion SasX had an influence in aggregation of S.aureus and its adherence to human nasal epithelial cells.By acquiring sasX,S.aureus colonized more easily to the susceptible sites of the host,and thus caused infection.

OBJECTIVETo evaluate the value of magnetic resonance imaging (MRI) in displaying the parotid gland segments of the facial nerve.

METHODSSixteen volunteers (9 males and 7 females) and 132 surgically confirmed patients with parotid tumors locating in the deep or shallow lobe of the parotid gland (including 89 with benign and 43 with malignant tumors) underwent MRI using T1WI and T2WI. The transverse images were obtained with the plane tilted 35 degrees to the foot, and the coronal images were acquired using conventional scanning.

RESULTSOn transverse T1WI, the parotid gland segments of the facial nerve displayed low signal with arc-shaped curve in the cross-section, showing a symmetrical dot-like low signal in the coronal plane. The facial nerve in 63% of the patients with parotid tumors in the cross-section could be displayed, but in the coronal plane the proportion reached 83%. MRI could accurately reveal the position of the parotid tumors in the deep or shallow lobe of the parotid gland.

CONCLUSIONMRI can show the major portion of the parotid gland segments of the facial nerve and has important value in locating the parotid tumors and displaying the relationship between the tumor and facial nerve.

Adolescent ; Adult ; Aged ; Facial Nerve ; pathology ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Parotid Gland ; innervation ; pathology ; Parotid Neoplasms ; pathology ; Young Adult

OBJECTIVETo explore whether fractional anisotropy (FA) value could be taken as a quantitative indicator for tracing and reexamining amyotrophic lateral sclerosis (ALS), and to analyze the correlation between FA value and integrative medical treatment.

METHODSTotally 18 ALS patients were recruited in this study. All patients received diffusion tensor imaging (DTI) using 3. OT (Propeller HD) MRI twice. Six regions of interest (ROI) were selected to measure FA values. Survival analyses were performed in 11 cases of end point events.

RESULTS(1) Three ROI (cerebral peduncle, posterior limb of internal capsule, and corona radiata) all indicated that FA value was the highest in patients with mild health status scale of amyotrophic lateral sclerosis (ALS/HSS). (2) There was statistical difference in the means of FA values in cerebral peduncle, posterior limb of internal capsule, and corona radiata of 18 cases between initial examination and reexamination (P < 0.01). (3) Kaplan-Meier survival curve showed the survival rate of ALS patients decreased as time went by, with the median survival time of 48 months.

CONCLUSIONSFA value was inversely proportional to the severity of ALS, the more severe, the lower FA values. FA value was an objective indicator for assessing the severity of ALS. ALS is an incurable disease till now. Integrative medical treatment might become one direction for ALS patients.

Amyotrophic Lateral Sclerosis ; diagnosis ; therapy ; Anisotropy ; Diffusion Tensor Imaging ; Humans ; Integrative Medicine

OBJECTIVETo investigate the apoptosis induced by manumycin in U937 and HL-60 cell lines, and to explore the role of mitochondria apoptotic pathway in manumycin-inducing apoptosis.

METHODSLeukemic cells line U937 and HL-60 were treated by manumycin at 2 micromol/L for different time. Apoptosis of leukemia cells was detected by flow cytometry. The cytosolic proteins were extracted using a digitonin buffer. The protein expression of cytochrome C, caspase-9, caspase-8, and caspase-3 were determined by western blot. Mitochondrial membrane potential was detected by JC-1.

RESULTSIn U937 and HL-60 cells, manumycin induced mitochondrial depolarization after 6 h treatment. The average red/green fluorescence ratios at 6 h were significantly (P < 0.01) lower than those at time 0, being 0.51 +/- 0.07 and 0.41 +/- 0.06 for control group respectively. Manumycin induced cytochrome C release from the mitochondria into the cytosol after 6 h treatment, and activated caspase-9, caspase-8, and caspase-3 after a 16h treatment. The broad-spectrum caspase-inhibitor Z-VAD-fmk at 50 micromol/L was able to inhibit caspase cleavage completely, but only reduced the manumycin-induced apoptosis rates by 51.69% and 56.47% in U937 and HL-60, respectively.

CONCLUSIONManumycin induced apoptosis in U937 and HL-60 cell lines via mitochondria apoptotic pathway.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cytochromes c ; metabolism ; HL-60 Cells ; Humans ; Mitochondria ; drug effects ; metabolism ; physiology ; Polyenes ; pharmacology ; Polyunsaturated Alkamides ; pharmacology