Objective With fat-soluble ingredients and water-soluble components as indicators,to investigate the processing technology of wine Gansu Salvia by orthogonal test.Method With fat-soluble ingredients and water-soluble components determined by HPLC,the processing technology of wine Gansu Salvia was optimized by orthogonal test.Result The transfer rate of fat-soluble ingredients with water extraction was 2.4%,with little significance,while it had a significant impact on salvianolic acid B.The best process condition was 10% yellow rice wine,added 20% wine,fried in 80 ℃ for 4 min.Conclusion It was reasonable to choose the best process condition for wine Gansu Salvia.

Objective To study the distribution of the gap conjunction and connexin(Cx)26 and 32 in inner ear of guinea pig. Methods We investigated expression and distribution of gap conjunction and Cx 26 and Cx 32 in cochlea of guinea by immunohistochemistry, RT-PCR and electron microscopy. Results T The gap conjunction was easily seen between hair cells and supporting cells in cochlea of guinea pigs. Cx26 was widely distributed in the stria vascularis, the spiral ligament and between the supporting cells, and Cx32 appeared mainly among supporting cells, in the stria vascularris. Conclusion There exist lots of gap conjunctions in inner ear of guinea pig. Two kinds of connexins can be seen in guinea pig's cochlea, and the Cx 26 plays more important role than other Cx.

Objective:To establish a method to determine the content of chlorogenic acid and rutin in nettle to assess the quality of nettle.Methods:An HPLC method was used with the following chromatographic conditions:CAPCELL PAK C18 column (250 mm × 4.6 mm, 5 μm) was used with the mixture of 0.4%phosphoric acid and methanol as the mobile phase with gradient elution , the de-tection wavelength was set at 340 nm, the flow rate was 1.0 ml· min-1 and the sample size was 10μl.Results:Chlorogenic acid with-in the range of 2.360-23.600μg · ml-1 showed a good linear relationship (r=0.999 9), and rutin within the range of 6.208-62.080μg · ml-1 showed a good linear relationship (r=0.999 9).The recovery of rutin and chlorogenic acid was 99.20% and 100.40%with RSD of 1.2%and 1.1%(n=6), respectively.Conclusion:The method is fast, effective, simple, accurate and reproducible , which can be used to quantitatively analyze chlorogenic acid and rutin in nettle .

Objective Rheumatoid arthritis (RA) is a typical type Ⅲ hypersensitivity with a large number of immune complexes (IC) and complement deposits in the synovial tissue , but its specific pathogenesis is not yet clear.This article was to explore the expression of the antigenic profile of serum ICs in RA.Methods ICs were isolated from the serum of 55 patients with RA (41 cases of anti-CCP antibody [+] and 14 cases of anti-CCP antibody [-]), 41 with systemic lupus erythematosus (SLE), and another 41 healthy controls by polyethylene glycol (PEG) precipitation, separated by immunoprecipitation, digested with trypsin in gel, and then subjected to mass spectrometry for identification.The levels of total proteins were compared among different groups using Vennny 2.1.0.The protein expression was considered to be up-regulated when the total protein level of the RA group was >2 times and down-regulated when it was <0.5 times that of the control.Further functional analysis was performed on the differential proteins in RA using the STRING software.Results Totally, 277 proteins were identified in the serum ICs of the RA patients, including 162 in the anti-CCP (+) and 248 in the anti-CCP (-) RA group.Compared with the SLE and healthy control groups, only 129 proteins were found in the RA patients, including 38 in the anti-CCP (+), 109 in the anti-CCP (-) RA group, and 18 in both the two groups.Among the proteins identified in the RA patients and healthy controls, 2 and 11 were up-regulated while 17 and 21 down-regulated in the anti-CCP (+) and anti-CCP (-) RA group, respectively.Conclusion More differentially expressed proteins were identified in the anti-CCP (-) than in the anti-CCP (+) RA patients.The identification of differentially expressed proteins provides a new idea and direction for the investigation of the pathogenesis and new biomarkers of RA.

60 curved root canals of permanent teeth with angles of curvature ranging from 15°to 40°(Schneider's methodology)were prepared using the instruments of Hyflex CM(HC) and ProTaper(PT) Universal respectively(n=30).Using standardized pre-and post-instrumentation paralleling periapical radiographs,canal curvature was determined by image analysis software and the clinical shaping effect of Hyflex CM and ProTaper rotary NiTi files were compared.The canal curvature in group HC and PT decreased by 4.54°±3.25° and 5.63°±3.84° respectively(between pre-and post-treatment in both groups,P<0.05;between groups,P>0.05).Hyflex CM can meet the clinical necessity for the instrumentation of curved root canals.

Objective The inhibitor of differentiation 3 (Id3) is an important transcriptional regulation factor, which participates in tumorigenesis, cell proliferation, and cell apoptosis.β-catenin, as a central molecule of the Wnt signaling pathway, is critical for tumor development.This study aimed to evaluate the expressions of these two molecules and the regulatory effect of Id3 on β-catenin in different tumor cells.Methods Total RNA was extracted using the Trizol Reagent.The relative mRNA expression levels of Id3 and β-catenin in tumor cells were detected by quantitative real-timePCR(qRT-PCR).The recombinant eukaryotic expression vector pEGFP/Id3 with the human Id3 gene was transfected into A549, A549/ DDP and SW-480 cells using the non-liposome-mediated method.The protein expressions of Id3 and β-catenin were determined by Western blot.Results The expression of Id3 was significantly lower in the colorectal cancer cell lines SW-480 and HT-29 than in A549 and other tumor cells (P<0.05), but remarkably higher in nasopharyngeal carcinoma CNE and 5-8F cells than in other tumor cells (P<0.05).The expression of β-catenin was the highest in SW-480in comparison withother malignant tumor cells(P<0.05), and the second highest was in gastric cancer AGS and colorectal cancer HT-29 cell lines, but low in H446, A549, SPC-A-1, A549/DDP, and SK-MES-1 cell lines and extremely low or almost absent in CNE and 5-8F cells (P<0.05).After transfected with pEGFP/Id3, the cells showed a decreased volume, wrinkled membrane and absent refraction under the fluorescence microscope, which, however, were not observed in most of the cells transfected with the empty vector pEGFP.Compared with the control, the Id3/pEGFP group showed remarkably increased expressions of Id3 mRNA in the A549, A549/DDI, and SW-480 cells (1.24±0.12 vs 193.12±2.80, 1.09±0.11 vs 188.30±2.60, and 0.92±0.29 vs 19.08±0.59, P<0.01), and the expression of β-catenin was significantly down-regulated in the transfected SW-480 cells with an overexpression of Id3 (0.98±0.05 vs 0.32±0.03, P<0.01), but exhibited no statistically significant differences from those in the transfected A549 and A549/DDP cells (0.98±0.07 vs 1.04±0.08 and 0.98±0.05 vs 0.32±0.03, P>0.05).Western blot showed the same results.Conclusion The expression levels of Id3 and β-catenin vary in different tumor cell lines.Anabnormally high level of β-catenin is an important risk factor for colorectal cancer, and the down-regulatedexpression of β-catenin after eogenous transfection of Id3 may provide some new ideas for target gene therapies of colorectal cancer.

OBJECTIVETo study the single-nucleotide substitution (c.-16C to T) of the PURATROPHIN-1 gene in spinocerebellar ataxia (SCA) patients in China.

METHODSThe single-nucleotide substitution (c.-16C to T) of the PURATROPHIN-1 gene was detected by PCR, digested with EcoN I, separated on 8% polyacrylamide gel in 68 probands of autosomal dominant SCA families and 119 sporadic SCA patients, who had been excluded CAG/CAA repeat expansion at the SCA1, 2, 3, 6, 7, 17 and dentatorubral-pallidolluysian atrophy (DRPLA) loci. The results were confirmed in four patients by direct sequencing.

RESULTSThe single-nucleotide substitution (c.-16C to T) of the PURATROPHIN-1 gene was not identified in authors' cohort.

CONCLUSIONThe mutation of c.-16C to T of the PURATROPHIN-1 gene might be rare in SCA patients in China.

Asian Continental Ancestry Group ; genetics ; Cohort Studies ; Guanine Nucleotide Exchange Factors ; genetics ; Humans ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Spectrin ; genetics ; Spinocerebellar Ataxias ; genetics

Objective To scan protein expression profile of immune complexes (ICs) derived from the synovial tissue of the patients with rheumatoid arthritis (RA) based on liquid chromatography-tandem mass spectrometry (LC-MS).Methods The samples of synovial fluid were obtained from knee joints of the patients with RA and osteoarthritis (OA) used as control during therapeutic arthrocentesis in knee jiont at the Department of Orthopedics of Jinling Hospital,School of Medicine,Nanjing University.The protein expression profile of ICs was identified by enrichment strategy based on immunoprecipitation and LC-MS analysis.The value of fraction of total (FOT) was used to estimate protein abundance and screen the up-and down-regulated proteins.The function enrichment,interaction network and signal pathway of differential proteins were analyzed using softwares David and String.Results A total of 511 and 526 protein spots in ICs of RA and OA patients were identified respectively.Among them,170 proteins existed only in RA group.45 and 85 proteins in RA group were statistically up-and down-expressed compared with controls.Conclusion HSP90AA1,HSP70,HLAG,Thioredoxin,Annexin A2 and vitronectin may be involved in the pathogenesis of RA through different paths and possible to become promising diagnostic indicators or new therapeutic targets for RA.

The remodeling process of synapses and eurotransmitter receptors of facial nucleus were observed. Models were set up by facial-facial anastomosis in rat. At post-surgery day (PSD) 0, 7, 21 and 60, synaptophysin (p38), NMDA receptor subunit 2A and AMPA receptor subunit 2 (GIuR2) were observed by immunohistochemical method and emi-quantitative RT-PCR, respectively. Meanwhile, the synaptic structure of the facial motorneurons was observed under a transmission electron microscope (TEM). The intensity of p38 immunoreactivity was decreased, reaching the lowest value at PSD day 7, and then increased slightly at PSD 21. Ultrastructurally, the number of synapses in nucleus of the operational side decreased, which was consistent with the change in P38 immhnoreactivity. NMDAR2A mRNA was down-regulated significantly in facial nucleus after the operation (P00<0.05), whereas AMPAR2 mRNA levels remained unchanged (P>0.05). The synapses innervation and the expression of NMDAR2A and AMPAR2 mRNA in facial nucleus might be modified to suit for the new motor tasks following facial-facial anastomosis, and influenced facial nerve regeneration and recovery.

OBJECTIVE: To explore the survival rate of neural stem cells (NSCs) from fetal rat and its biological properties after cryopreservation and thawing. METHOD: Different generations NSCs (the first generation, the third generation and the sixth generation) from fetal rat which cultivated with serum-free medium in vitro were cryopreserved in the cryogen, which were neurosphere culture medium with 10% BSA and 7.5% DMSO (without neural growth factor). The cryopreserved cells were resuscitated at 1st week, 4th week, 8th week, 12th week and 16th week respectively. The survival rate of cells were calculated and the cells were incubated and differentiated again. RESULT: Different time of cryopreservation, different generations did not affect NSCs survival (P > 0.05) after cryopreservation. The survival rate of NSCs was from 60% to 70% after resuscitated, which were differentiated into neurons and astrocytes in 10% embryonic bovine serum (without growth factor). CONCLUSION NSCs were successfully cryopreserved, resuscitated and recultured, which would create bases for the experimental study on the selected-date application of NSCs transplantation into cochlea for treating sensorineural deafness.
Animals ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Proliferation ; Cell Survival ; Cells, Cultured ; Cryopreservation ; Female ; Fetus ; cytology ; Neural Stem Cells ; cytology ; Neurons ; cytology ; Pregnancy ; Rats ; Rats, Sprague-Dawley