Objective To establish a rapid test method for the determination of five sweeteners and two preservatives in the milk beverage by HPLC.Methods Trough a set of pretreatment such as dilution,protein precipitation,high-speed centrifugation,the samples were tested.With NUCLEODUR C18 RP-column separation,phosphate buffer(pH=5.0) and acetonitrile were used as the mobile phase,gradient elution analysis was conducted,the detective wavelength was 205nm.Results The separation of one sample was achieved within 15 min,five sweeteners and two preservatives could be well separated,the linear range was 5.00-100.0 mg/L,r≥0.9993,RSD was 1.5%-4.2%(n=6),the average recovery rate was 85.2%-108%.Conclusion This method is simple and quick with better reproducibility and selectivity,it is an effective way to determine sweeteners and preservatives in milk beverage.

Objective:To study the relationship between serum soluble intercellular adhesion molecule-1(sICAM-1) and vascular endothelial growth factor(VEGF), placenta growth factor(PLGF), vascular endothelial growth factor receptor 1(VEGFR1, Flt-1), soluble vascular endothelial growth factor receptor 1(sVEGFR1, sFlt-1) mRNA expression in placenta tissue of preeclampsia(PE). Methods: The serum level of sICAM-1 was measured by enzyme-linked immunosorbent assay and gene expression of placenta tissue was detected by quantitative reverse transcriptase-polymerase chain reaction(RT-PCR). Results:(1)The average serum level of sICAM-1 was(218.45±29.93) μg/L in PE group compared with (168.84±19.39) μg/L in controls(P < 0.01).(2)The mRNA expressions of VEGF, PLGF, Flt-1 and sFlt-1 were increased in placenta of PE than those in controls (P < 0.01).(3)There was a positive correlation between the serum level of sICAM-1 and the sFlt-1 mRNA expression in placenta tissue(r = 0.90, P < 0.01). Conclusion: The serum level of sICAM was increased in the patients with PE. The expressions of VEGF, PLGF, Flt-1 and sFlt-1mRNA were increased remarkably in the placenta tissue of palients with PE,especial for sFlt-1. The remarkable increase for expression of sFlt-1mRNA may contribute to endothelial dysfunction, hypertension and proteinuria.

Myelin of the adult mammalian central nervous system(CNS)has been attributed to affect nerve structural plasticity and suppress regeneration of nerve fibers.Nogo-A is possibly the best characterized of a variety of neurite growth inhibitors in CNS myelin.Neutralizing its activity results in improved axon regrowth and functional recovery in experimental spinal cord injury(SCI)models of animals.Nogo-A and its receptors,especially Nogo-66 receptor(NgR),p75 neurotrophin receptor(p75NTR),and LINGO-1 increasingly become the hot spot in the study of SCI repair,and have become the major targets for therapeutic intervention to promote axon regeneration after SCI.Inhibition of Nogo-A and its receptors NgR/p75NTR/LINGO-1 may be promote the regeneration of axon and maximize functional recovery after SCI.

Objective This study was designed to establish and optimize the two-dimensional gel electrophoresis (2-DE) for the proteome analysis of human Transitional Cell Carcinomas of the Urinary Bladder ,and to established the two-dimensional gel electrophoresis (2-DE) profile of human Transitional Cell Carcinomas of the Urinary Bladder(TCC) of different stages in carcinogenic process. The basic aim is to perform differential analysis and provide a basis for identifying carcinogenesis-associated protein of TCC.Methods After obtaining samples of the normal,invasive tissue,noninvasive tissue,a series of methods Such as sample preparation,electrophoresis parameters,gel concentration,PDQuest 2DE analysis software,peptide mass fingerprint (PMF,MALDI-TOF-MS)were used to analyses these tissues.Results The good 2DE patterns of different tissues including resolution and reproducibility were obtained. After coomassie Brilliant Blue R-250 staining, the average matching ratio was 76%.There was a good reproducibility of spot position in 2-DE map, with average deviation in IEF direction of (0.98?0.26) mm, while in SDS-PAGE direction it was(1.23?0.22) mm. The 2-DE patterns with good quality had been obtained. Ten protein spots chosen randomly from different stages were identified by PMF, some of which were involved in the cell proliferation, differentiation, cycle regulation, and tumor occurrence, such as Heat shock protein-27,SFN etc.Conclusions The results provide a fundamental basis for further study of mechanism of human Transitional Cell Carcinomas of the Urinary Bladder and screening its specific marker.

Objective To study the expression of MCM5 gene in urine exfoliate cell and bladder cancer tissue in order to research the diagnosis and difference. Methods We collected the samples of urine exfoliate cell and bladder cancer tissue and extracted the total RNA, and then did RT-nest-PCR, immunohistology to check the expressive level of mcm5 gene. Results Sensitivities of that are 93。3% in 30 bladder cancer, 13。3% in other patients and 0% in normal person. The expression of mcm5 between G1?G2 and G3 bladder cancer tissue have very marked statistic significance ( P

Objective To establish a method of semiquantitative polymerase chain reaction for detecting cdc6 gene in EJ cells and bladder cancer tissue.Methods We collected the samples of urine EJ bladder cancer cells and extracted the total RNA, and then did RT-nest-PCR to check the expressive level of cdc6 gene. Result We established stable semiquantitative PCR by putting primer of beta-actin served as internal reference gene and primer of target gene in the same test tube and optimizing experiment parameters for PCR, such as the concentration of magnesium and cycle times etc. The intro and inter group CV were 8.01 and 14.53 respectively for cdc6. The detecting limit was 5?10 -2 ng.Conclusion It is usable to test the expressive level of cdc6 gene in bladder cancer.

Objective To investigate the effects of alcohol extract of Kunmu decoction on proliferation and apoptosis of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS). Methods Synovial tissues were obtained from patients with active RA received joint replacement or arthroscopy. The surface antigen and the amount of apoptotic cells were determined by flow cytometry. The inhibitive effect was detected by MTT assay. Results The CD90+surface antigen of synoviocytes was (94.78 ± 0.98)%. The inhibitive effect on the proliferation in all treatment groups were in a time-and dose-dependent manner. The apoptosis rate was increased in a dose-dependent manner among all dosage alcohol extract groups. Conclusion Kunmu decoction might inhibit proliferation and induce apoptosis of RA-FLS.

Objective To obtain differentiated osteoblast-specific inactivation of fgfr1 mice Methods To obtain fgfr1△/+/OC-CreTG/+ mice,fgfr1flox/flox mice obtained from NIH were crossed with OC-Cre mice To obtain fgfr1△/△/OC-CreTG/+ mutant mice,fgfr1△/+/OC-CreTG/+ further crossed with themselves or fgfr1flox/flox mice After fgfr1△/△/OC-CreTG/+ crossed with fgfr1flox/flox mice,half of their offspring were mutant mice Results Differentiated osteoblast-specific fgfr1 knockout mice were obtained Conclusion fgfr1△/△/OC-CreTG/+ mice were obtained through proper crossing strategy,which provides a suitable platform for studying fgfr1 function in bone development and fracture healing

Humans ; Sodium-Calcium Exchanger ; Carrier Proteins ; Pain ; Calcium/metabolism*

OBJECTIVE： To establish HPLC fingerprints of Paeonia tactilora decoction pieces， and to conduct its cluster analysis and principal component analysis. METHODS： HPLC method was adopted. The determination was performed on SunFire® C18 column with mobile phase consisted of acetonitril-0.05％ phosphoric acid solution （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 230 nm， the column temperature was 30 ℃， the collection time was 70 min，and sample size was 15 μL. Using paeoniflorin as reference， HPLC fingerprints of 26 batches P. tactilora decoction pieces from different habitats and 30 batches by different processed methods were established. The similarity of samples was evaluated by TCM Chromatographic Fingerprint Similarity Evaluation System （2012 edition） to confirm common peak. Cluster analysis and principal component analysis were performed by using SPSS 20.0 software. RESULTS： There were 9 common peaks in HPLC fingerprints of 26 batches of sample from different habitats， the similarity of which was higher than 0.880. Six peaks were identified， including gallic acid， catechin， albiflorin， paeoniflorin， 1，2，3，4，6-pentagalloylglucose and benzoylpaeoniflorin. Cluster analysis showed that 26 batches of samples were clustered into 2 categories when cosine distance was 15. S1-S21 were clustered into one category； S22-S26 were clustered into the other category. By principal component analysis， the accumulative contribution rate of two main components was 81.124％. There were 10 common peaks in HPLC fingerprints of 30 batches of sample by different processed methods， the simi- larity of which was higher than 0.970. Seven peaks were identified， including gallic acid， catechin， aplopaeonoside， albiflorin， paeoniflorin， 1，2，3，4，6-pentagalloylglucose and benzoylpaeoniflorin. Cluster analysis showed that 30 batches of samples were clustered into 2 categories when cosine distance was 25. B1-B10 were clustered into one category； C1-C10 and J1-J10 were clustered into the other category. By principal component analysis， the accumulative contribution rate of four main components was 86.887％. CONCLUSIONS： Established HPLC fingerprint， the results of cluster analysis and principal component analysis can provide reference for quality control of decoction pieces of P. tactilora.