OBJECTIVE:To establish the UPLC fingerprint for Qinghou liyan granule,and provide reference for its quality control by combining with principal component analysis (PCA). METHODS:UPLC was performed on the column of ACQUITY UPLC BEH C18 with mobile phase of acetonitrile(A)-0.1% aqueous phosphoric acid solution(B)(gradient elution)at a flow rate of 0.4 mL/min,the detection wavelength was 280 nm,column temperature was 30℃,and injection volume was 2μL. Using ellag-ic acid as a reference,12 batches of samples were analyzed,Similarity Evaluation Software for Chromatographic Fingerprint of Traditional Chinese Medicinewas used for the similarity analysis and identification of the common peaks,and the PCA was used for common peaks. RESULTS:There were 18 common peaks in the fingerprints of 12 batches of samples,and 6 principal peaks (gallic acid,ellagic acid,naringin,hesperidin,neohesperidin and baicalin) were identified;the similarity degree of 12 batches and reference fingerprints were no less than 0.984. According to PCA,the 18 peaks can be integrated into 3 principal components, with cumulative contribution rate of principal component of 78.277%;baicalin and 16 peaks were the discriminating factors of the fingerprint of Qinghou liyan granule. CONCLUSIONS:The method can provide reference for the quality control of Qinghou liyangranule.

Objective To optimize ultrasonic-assisted extraction Epimedin C and Icariin from Epimedii Wushanensis Herba by response surface methodology. Methods On the basis of single factor tests, Box-Behnken experimental design and response surface methodology were adopted to optimize extraction conditions with the concentration of ethanol, ultrasonic time and solid-liquid ratio as factors. HPLC was used to determine the content of Epimedin C and Icariin from Epimedii Wushanensis Herba. Results Optimal ultrasound-assisted extraction technology was as following: the concentration of ethanol was 73%; the ultrasonic time was 22 min; the ratio of liquid to material was 1:32. The contents of Epimedin C and Icariin from Epimedii Wushanensis Herba were 15.90% and 0.75%, respectively. Conclusion This extraction technology can improve the extraction efficiency of Epimedin C and Icariin from Epimedii Wushanensis Herba, which is in accordance with predicted value.

OBJECTIVE:To establish a method for the contents determination of gallic acid,naringin,hesperidin,neohesperi-din and baicalin in Qinghou liyan granule. METHODS:UPLC was performed on the column of ACQUITY UPLC BEH C18 with mobile phase of acetonitrile- 0.1% phosphoric acid(gradient elution)at a flow rate of 0.40 ml/min,detection wavelength was 280 nm,column temperature was 30 ℃,and injection volume was 1 μl. RESULTS:The linear range was 0.048-0.480 μg for gallic ac-id(r=0.999 3),0.012-0.120 μg for naringin(r=0.999 9),0.016-0.160 μg for hesperidin(r=0.999 8),0.022-0.220 μg for neo-hesperidin(r=0.999 9)and 0.072-0.720 μg for baicalin(r=0.999 2);limit of quantitation was 2.4,1.6,1.8,1.8,1.6 ng,limit of detection was 7.5,5.0,5.6,5.6,5.0 ng;RSDs of precision,stability and reproducibility tests were no lower than 3.0%;recover-ies were 96.64%-102.02%(RSD=2.00%,n=6),95.86%-102.56%(RSD=2.86%,n=6),97.24%-102.54%(RSD=2.10%,n=6),97.44%-102.60%(RSD=2.40%,n=6)and 96.91%-103.13%(RSD=2.62%,n=6). CONCLUSIONS:The method is rapid and accurate,and can be used for the simultaneous determination of gallic acid,naringin,hesperidin,neohesperidin and baicalin in Qinghou liyan granule.

Objective To study the effects of closed self-help sterile gloves wearing in surgical operations.Methods One hundred surgical personnel were randomly divided into two groups:the observation group(n=51)using the closed self-help gloves wearing and the control group(n=49)using the conventional wearing.The two group were compared about the rates of edge curling of the gloves and cuff exposure.Result The closed self-help gloves wearing group had significantly lower rates of edge curling and cuff exposure than the control group(P<0.05,for both).Conclusion Closed self-help gloves wearing can effectively prevent the edge from curling during wearing gloves and reduce cuff exposure in the operation so as to reduce the risk of surgical contaminations.

Aim To investigate the effects of quercetin and psoralen on the proliferation of human breast carcinoma cells in vitro.Methods Effects of quercetin and psoralen on the cell proliferation was tested in ER-positive MCF-7 cells by flow cytometer and Western blot.And the estrogen-like effect of psoralen and its relation with the estrogen-receptor were evaluated by pure estrogen receptor antagonist ICI182,780 as a tool.Results ① Psoralen(10 ?mol?L-1) and quercetin(10 ?mol?L-1) stimulated proliferation of MCF-7 cells compared with vehicle control,and the cell cycle was impulsed from G1 to S,DNA synthesizing was enhanced.It was also found the above function on boosting MCF-7 cell proliferation could be inhibited by adding estrogen receptor antagonism ICI182,780.② Psoralen(10 ?mol?L-1) and quercetin(10 ?mol?L-1) up-regulated ER? protein levels without altering ER? levels.The above up-regulation on ER? protein could be inhibited by adding estrogen receptor antagonism ICI182,780.Conclusions Psoralen and quercetin have the estrogen-like activities through the estrogen response pathway.And they exert estrogenic activity to MCF-7 cell proliferation through interaction with ER? expression.

AIM:To characterize the inclusion complex of volatile oil of Rhodiola crenulata/?-cyclodextrin. METHODS: Some analytical methods,such as DTA,IR,GC,TLC were apllied to the investigation before and after the inclusion. RESULTS: The difference between the inclusion and physical mixture in differential thermal(analysis,) infrared spectra,gas chromatogram and thin layer chromatogram. CONCLUSION: The inclusion of Rhodiola crenulata volatile oil has the characteristic of intramolecular inclusion.

BACKGROUND: An ideal marked molecule has not been found to detarmine bone marrow mesenchymal stem cells (MSCs) so as to make sure the homogenicity.OBJECTIVE: To verify the in vitro differentiation from BrdU-treced MSCs into osteoblasts.DESIGN, TIME AND SETTING: A cytological observation/n vitro was performed in Shanxi Medical University from January to October 2008.MATERIALS: Wistar rats aging 4 weeks old were provided by Experimental Animal Center of Shanxi Medical University.METHODS: MSCs were isolated and cultured by using density gradient cantrifugation combined with attachment culture method.At about 80% confluence, trypsin was used for passage and amplification. MSCs at density of 5×1010/L were inoculated in a 25-mm culture dish with L-DMDM culture medium containing dexamethasone, β -phosphoglycarol, vitamin C, and 10% fetal bovine serum. The third-passaged MSCs were labeled in vitro with 10 μmol/L BrdU tracer. Thereafter, 10 visual fields were randomly selected to calculate numbers of positive and negative ceils so as to obtain BrdU tracing rate under a fluorescence microscope (×200).MAIN OUTCOME MEASURES: Inverted microscope was used to observe cell morphology; flow cytometry was used to detect cell surface antigen, differentiation into osteoblasts, and BrdU tracing rate in vitro.RESULTS: The purified MSCs which were like fibroblasts were adherent and fusiform. The third-passaged cells were changed equidirectionally and whirlpool-arranged, and the survival rate was more than 95%. The seventh-passaged cells still grew rapidly.CD44, CD71, and CD105 expressions were positive, but CD45 expression was negative. Black particles were visualized in MSCs after Von kossa staining. BrdU tracing rate was more than 90%.CONCLUSION: Density gradient centrifugation combined with attachment culture method can effectively isolate and purify rat MSCs which are cultured in vitro for a long period and differentiated into osteoblasts. BrdU tracer is safe, effective, and convenient to successfully label MSCs.

Objective: To understand the relationship between childhood psychological resilience and of school violence awareness in rural areas in southern Henan, and to provide support and reference for the prevention and control of school violence. Methods: A total of 6 484 primary school children in grades 4-6 in the rural areas of Dengzhou, Xincai and Gushi counties in southern Henan were surveyed for psychological resilience and awareness on school violence. Results: The average score of psychological resilience of grade 4-6 pupils in rural areas of southern Henan was (39.92±8.18), and the average score of school violence awareness was (60.78±6.19). In terms of age, gender, grade, school type, personality, class performance, academic performance, smoking, intimate friends, playing games, truancy, loitering outside the school, relationship with siblings, father s education, mother s education, parental conflict, parenting style, the proportion of primary school students in different level of school violence awareness varied statistically different (P<0.01). Multivariate Logistic regression showed, after controlling for confounding factors, psychological resilience was an independent influencing factor affecting awareness on school violence (OR=1.06, 95%CI=1.05-1.08), with better psychological resilience, being associated with higher school violence awareness (P<0.01). Conclusion The level of school violence awareness and psychological resilience of pupils in southern Henan is relatively low. The higher level of psychological resilience, the higher level of school violence awareness; psychological resilience might be improved through education and household parenting modification to prevent and reduce the incidence of violence on campus.

Objective To investigate the technical requirements for the 3.0T multi-modal quantitative MR imaging of the rabbit liver.Methods A total of 8 rabbits were scanned and T1 mapping,T2 mapping,MT and Gd EOB-DTPA dynamic enhancement images were acquired.For the dynamic enhanced scanning,injecting contrast medium was followed by saline flush with different combinations of injection flow rate (1.5 mL/s and 2 mL/s) and injection volume(6 mL and 8 mL).The quality of the images was assessed and analyzed statistically.Results The total scanning time was about 20-25 min.High-quality images were acquired by T1 mapping,T2 mapping and MT sequences.There was no significant difference in image quality among different groups in Gd-EOB-DTPA dynamic enhanced scanning(P＞0.05).Two rabbits died when the combination of injection flow rate of 2 mL/s and injection volume of 8 mL was used.Conclusion 3.0 T multi-modal quantitative MR scanning of rabbit liver can be achieved successfully if scanning parameters are properly chosen.

BACKGROUND:Spinocerebel ar ataxia type 3 (SCA3) is a typical genetic neurodegenerative disease. To establish patient-specific disease models of genetic background contributes to studying the pathogenesis and exploring therapeutic manners.
OBJECTIVE:To observe the effectiveness of neural differentiation of induced pluripotent stem cel lines induced by SCA3 and the stability of CAG copy number.
METHODS:Skin tissue of SCA3 patient was obtained clinical y, and specific skin flbroblasts were isolated and cultured. Reprogramming fibroblasts could obtain induced pluripotent stem cel s. Patient-specific induced pluripotent stem cel s, normal person induced pluripotent stem cel s (NHF) and embryonic stem cel s (ES-10) were induced to differentiate. Flow cytometry was used to compare the efficiency of differentiation. Western blot assay was utilized to detect ataxin-3 protein expression in neurons. Polymerase chain reaction was applied to measure the CAG repeat number of SCA3/ATXN3 gene.
RESULTS AND CONCLUSION:Induced pluripotent stem cel s that had identical genetic background to fibroblasts were successful y obtained, and had similar morphology and multi-directional differentiation potential to human embryonic stem cel s. Each cel line could differentiate into neural stem cel s. The CAG number did not apparently alter before and after reprogramming as wel as induction of neuronal differentiation. The effectiveness of the differentiation of induced pluripotent stem cel s derived from SCA3 into neural stem cel s was lower than that of normal person-derived induced pluripotent stem cel s (NHF) and embryonic stem cel s (ES-10). These findings demonstrate that reprogramming can successful y establish human induced pluripotent stem cel s, and induced the differentiation of above cel s into neural stem cel s. In the whole process, CAG number did not obviously alter, which was consistent with body cel s of patients.