Objective To development a cooling crystallization process that is suitable for industrial preparation of purified dihydromyricetin.Methods Screen design was used to investigate effects of process parameters such as,temperature,concentration ethanol aqueous,quantity of activated charcoal and adsorption time on yield and purity of dihydromyricetin.Purity was verified by high performance liquid chromatography and thin layer chromatography.The solubility of dihydromyricetin in water at viable temperature and ethanol proportion was also determined by UV spectrophotometry.The solid form was characterized by thermal analysis and powder X-ray diffractometry.Results When temperature was ＞ 85 ℃,ethanol concentration ＜ 10％,dosage of activated charcoal 0.1％-0.3％,and adsorption time 1-3 min,yield of dihydromyricetin was more than 70％,and the purity greater than 98％.The crystals,prepared by cooling crystallization from water and ethanol aqueous,had the same physical form and crystal habit.Conclusion Cooling crystallization from low concentration of ethanol aqueous gets higher yield and the process is more robust than crystallization from water.

BACKGROUND:Pelvis is the most complicated structure of human body, and many scholars have devoted themselves to its biomechanical study. However, it is difficult to construct the pelvic finite element model containing main muscles and ligaments, as a result of its irregular shape and complicated structure.
OBJECTIVE:To establish a normal human pelvis three-dimensional finite element model and provide digital model for the subsequent biomechanics analysis of pelvis.
METHODPelvic CT images of healthy male volunteers were stored in DICOM format and then put into Mimics 10.01 software for three-dimensional medical image reconstruction. The files were imported into reverse engineering software Geomagic studio 11 for surface processing and establishing surface model;then the surface model was introduced into finite element analysis software Ansys 14.0 for finite element meshing.
RESULTS AND CONCLUSION:A normal adult pelvis three-dimensional finite element model was developed, including 818 294 units and 149 290 nodes. Based on the CT images, we established pelvic finite element model, which have good accuracy, anatomical and geometric similarity, and could provide the basis for subsequent biomechanics analysis of pelvis.

Objective To investigate the growth inhibition and DNA damage of energized fusion protein anti-CD20(Fab)-LDM on B JAB cells in vitro.Methods The binding activity of fusion protein anti-CD20 (Fab)-LDP to B JAB cells was studied by flow cytometry and confocal laser scanning microscopy.MTT assay was used to study the energized fusion protein anti-CD20(Fab)-LDM on cell growth of B JAB cells.Comet assay was employed to detect DNA damage in B JAB cells.The cell growth cycle of BJAB was analyzed by FACS.Results The recombinant fusion protein anti-CD20 (Fab)-LDP possessed an significant target affinity towarded BJAB cells.The energized fusion protein anti-CD20(Fab)-LDM showed obvious inhibition on proliferation,as well as induced potent DNA damage in B JAB cells in vitro compared with lidamycin.B JAB cells treated with energized fusion protein anti-CD20 (Fab)-LDM showed S phase cell cycle.Conclusion The energized fusion protein anti-CD20 (Fab)-LDM could target binding to BJAB cells and significantly inhibit the proliferation of B JAB cells by inducing DNA damage and S phase arrest.

Objective:To study the therapeutic effect of a novel double-target system,in which human umbilical cord-derived MSCs were used as vehicles to deliver fusion protein scFvCD20:sTRAIL to non-Hodgkin ’ s lymphoma. Methods: The traditional methods in molecular biology were used to construct lentivirus expression vectors pLenR. scFvCD20: sTRAIL and contrast vectors. Human umbilical cord-derived MSCs ( HUMSCs ) were labeled with the copGFP by transducing with pseudo viral particles which had been packaged in 293T cells with four plasmid-lentivirus packaging system. Fusion protein scFvCD20:sTRAIL were secreted from MSC. scFvCD20:sTRAIL after that HUMSCs were infected by pseudo viral particles. CCK8 assay was applied to detect the antigen-restricted cell death induced by scFvCD20:sTRAIL in CD20-positive BJAB and Raji cells as well as CD20-negtive Jurkat cells and human normal peripheral blood mononuclear cells (PBMCs). To evaluate the therapeutic effect of MSC. scFvCD20:sTRAIL in vivo,ge-netically modified HUMSCs were intravenously injected into tumor-bearing mice with BJAB cells. The volume of tumor was measured every three days, and the inhibition ratio of tumor was calculated according to tumor volume. Results: Lentivirus expression vectors pLenR. scFvCD20:sTRAIL, pLenR. ISZ:sTRAIL, pLenR. scFvCD20 and pLenR. CopGFP were successfully constructed and these constructs could be expressed stably in HUMSCs by lentivirus transduction. scFvCD20:sTRAIL fusion protein produced a potent inhibition of cell proliferation in CD20-positive BJAB cells,moderate inhibition of the growth of Raji cells,and weak inhibition in CD20-negtive Jurkat cells when compared with ISZ-sTRAIL treatment,and it had no effect on normal human peripheral blood mononuclear cells (PBMCs). The MSC. scFvCD20:sTRAIL treatment significantly inhibited the tumor growth when compared with those treated with MSC. ISZ-sTRAIL. Conclusion: A double-target therapeutic system is well established, in which HUMSCs migrated to tumor site, secreted a novel fusion protein scFvCD20:sTRAIL,and thus locally concentrated scFvCD20:sTRAIL extended antigen-restricted anti-tumor activity. The engineered HUMSCs secreting scFvCD20:sTRAIL showed potent effect on inhibiting tumor growth in BJAB lymphoma malignancy,which may play an essential role in the clinical research .

Objective To investigate the clinicopathologic features and prognostic significance of BRCA1 and Ki-67 expression in breast cancer.Methods The expression levels of BRCA1 and Ki-67 were assayed by immunohistochemistry in 194 cases of breast cancer tissues.The correlations of BRCA1 and Ki-67 expression with patients' clinicopathologic features were also analysed.Results Low expression of BRCA1 was detected in the lymph node metastasis group,ER/PR negative group,and HER-2 positive group in 194 patients with breast cancer (P ＜ 0.05),as well as in the triple negative breast cancer (TNBC) group compared to non-TNBC group (P ＜0.05).High expression of Ki-67 was detected in patients with higher histological grade,negative ER/PR,and positive HER-2 (P ＜ 0.05).Furthermore,negative correlation was found between the expression of BRCA1 and Ki-67 (P ＜ 0.05).The combination of low expression of BRCAl and high expression of Ki-67 was mostly found in the patients with postmenopausal,lower histological grade,lymph node metastasis,negative ER/PR and positive HER-2 (P ＜ 0.05).However,there was no significant difference between TNBC and non-TNBC.Conclusion Joint detection of BRCA1 and Ki-67 might play an important role in predicting clinical outcomes of breast cancer,especially BRCA1 may be one of prognostic factors in TNBC.

AIM: To clarify the effect of SC58125 on cell proliferation and apoptosis in HepG-2 cells and explore the molecular mechanisms. METHODS: Cell culture, MTT, TUNEL, DNA ladder, flow cytometry and Western blot analysis were employed in the present study. RESULTS: SC58125 inhibited the growth of HepG-2 cells and induced the apoptosis. Furthermore, it arrested G_0/G_1 phase and inhibited S phase in HepG-2 cells. Depressed expression of P33 cdk2 ,P34 cdc2 ,cyclin B_1,cyclin E ,Mpm-2,Rb ,PCNA proteins were found in HepG-2 cells treated with SC58125. CONCLUSION: SC58125 inhibits cell proliferation and induces apoptosis, which may be related to the altered low protein levels of P33 cdk2 ,P34 cdc2 ,cyclin B_1,cyclin E ,Mpm-2,Rb,PCNA

Objective To observe the protein expressions of PI3Kp110, pAkt, pGSK3β of phosphatidylinositol 3-kinase (PI3K) and protein kinase B (Akt) signaling pathway in the whole blood of patients with Kashin-Beck disease and analyze the status of PI3K/Akt signaling pathway. Methods Patients with Kashin-Beck disease ( KBD group ) were from six counties ( Xunyi , Linyou , Yongshou , Qianyang , Changwu and Long County) of Shannxi Province in Kashin-Beck disease areas , and the healthy controls (control group) were matched by age and sex. Venous blood was collected from patients and healthy controls. Trizol method was applied to extract the whole blood protein; protein expression levels of PI3K/Akt signaling pathway in whole blood were detected by Western blotting; the gray values were observed and recorded by the sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) and GDS-8000 gel imaging analysis system. Differences between the two groups were assessed by Student’s t-test. Results Compared age and sex between KBD group and control group, differences were not statistically significant(t=0.701, P>0.05;χ2=0.400, P>0.05). The protein expression levels of PI3Kp110, pAkt and p-GSK3β in KBD group were higher than that in control group(156.1 ± 92.1 vs. 79.5 ± 21.5, 113.7 ± 15.2 vs. 43.3 ± 10.7 and 105.9 ± 17.5 vs. 37.3 ± 12.0, respectively) and the differences were statistically significant (t=2.563, 6.567, 7.916; all P < 0.05 or < 0.01). Conclusion The PI3Kp110, pAkt and p-GSK3β expressions of signaling pathway proteins in the whole blood of patiens with Kashin-Beck disease are up-regulated significantly and the status of PI3K/Akt signaling pathway is activated.

Objective Although minimally invasive thoracoscopic surgery has unique advantages, there are still some difficulties in the complete resection of the thymus.Video-assisted thoracoscopic surgery has made up for the lack of thoracoscopy.This article summarize and analyze the clinical effects of DaVinci robotic surgery and thoracic surgery in the treatment of thymic diseases.Methods 116 cases of patients with thymic lesion were included in the study.According to the different treatment methods, 65 cases were included in da Vinci robotic group and other 51 cases were thoracoscopic group.Comparison was made in operating time, time of chest tube extubation, bleeding volume during surgery and amount of drainage within 24h after the operation between two groups.Results There were no statistical differences between robot group and thoracoscopic group in operating time(P>0.05).The time of pulling out the chest tube in robot group, postoperative days in surgical intensive care unit (SICU), and postoperative hospitalization days were significantly shorter than those in thoracoscopic group ([2.54±0.32]d vs [2.87±0.49]d, [0.75±0.04]d vs [0.81±0.06]d, [5.17±0.15]d vs [5.50±0.23]d, P<0.05).The bleeding volume during surgery and amount of drainage within 24h after the operation in robot group had a significant advantage over those in thoracoscopic group ([125.7±7.5]mL vs [136.6±8.2]mL, [117.38±11.45]mL vs [122.41±13.14]mL, P<0.05).Conclusion The da Vinci robotic surgery has the advantages of minimal trauma and rapid recovery, which makes up for the deficiency of thoracoscopy to a certain extent.

OBJECTIVEA novel method was established and the results were compared for HPLC fingerprint determination of crude and processed products of Entada phaseoloides.

METHODHPLC-UV was proposed for studying the fingerprints of fresh E. phaseoloides and their processed products, respectively. HPLC-ESI-MS was introduced to analyze the common peaks in each batch of crude E. phaseoloides.

RESULTSixteen characteristic peaks were found in crude E. phaseoloides samples and twenty-one common peaks existed in processed E. phaseoloides samples. Nine characteristic peaks of which were identified by comparison of the retention time and their molecular weights of chemical standards, most of which were identificated belong to triterpenoid saponins and glucosides.

CONCLUSIONAfter processed, the chemical composition of the extraction with solution of 60% methanol from crude E. phaseoloides are less or more than that from processed E. phaseoloides, and the changes of the main peaks of fingerprint chromatographic suggest that HPLC can be used to reflect the difference of chemical composition of E. phaseoloides and their processed products. It would be an efficient way for qualitative control of E. phaseoloides.

Objective To evaluate the accuracy and clinical application of endoscopic ultrasound guided fine needle aspiration (EUS-FNA) in diagnosis of occupying lesions in upper digestive tract and its pe-ripheral areas. Methods The data of 64 patients who underwent EUS-FNA for occupying lesions in upper di-gestive tract, between July 2006 and December 2007, were retrospectively analyzed. Results EUS-FNA was successfully performed on 62 patients, with a success rate of 96.88%. The overall diagnostic accuracy, sensi-tivity, specificity, positive predictive value and negative predictive value of EUS-FNA for occupying lesions in upper digestive tract and its peripheral areas were 88.71%, 86.54%, 100.00%, 100.00%, and 58.82%, respectively. There was no significant difference between the foei larger and smaller than 3cm (P > 0.05), re-garding the diagnostic accuracy and average puncture numbers. Diagnostic accuracy of those with real-time cy-tolngical evaluation was significantly higher than those without (P=0.029), and the puncture numbers were less (P=0.001). Among the total 99 punctures, the positive diagnostic accuracy of those with 5ml negative pressure suction was significantly higher than those of 10 ml (P = 0.044). Conclusion EUS-FNA is a safe and applicable approach to diagnosis of occupying lesions in upper digestive tract and its peripheral areas with higher diagnostic accuracy, sensitivity, specificity and positive predictive value. Moreover, the real-time cyto-logical evaluation and adequate negative pressure might facilitate the diagnosis accuracy.