OBJECTIVE:To establish RP-HPLC method for the content determination of praeruptorin A in Qingfei yihuo pills.METHODS:The chromatographic column was Diamonsil-C18(250 mm?4.6 mm,5 ?m) column.The mobile phase consisted of methanol-water(81:19) and the detection wavelength were set at 322 nm.RESULTS:The linear range of praeruptorin A was 4.65～23.25 ?g?mL-1(r=0.999 9) with an average recovery of 100.5%(RSD=2.35%,n=6).CONCLUSION:The method is sensitive,well separated and accurate for content determination of Qingfei yihuo pills.

Building a lean and efficient student league team should be paid attention to in the work of universities'youth league.This is decided by the characteristics of the knowledgeability of the youth leageue member,the mass character of the work scope and the cyclicality of the work process.The status quo of the current student league team is not satisfactory,so we should strengthen the building of this team from the selection and appointment system,the ideological management,the training of the operational capacity,the practice and standardization of system management,and the reinforcement of the assessment mechanism.

Objective To explore the expression of FLT3 on the cells from acute myeloblastic leukemia (AML)and its clinical signficance.Methods The expression of FLT3 from 80 AML cases was measured by FISH.All cases were indentified by karyotyping,40 cases were normal karyotye,and the other 40 cases were abnormal.Results The FLT3 expression level was different in different AML cases.The expression rate in normal karyotype cases was 5.0%,but in abnormal cases 30.7%,in refractory relapse cases 43.3%,in continual complete remission cases 4.5%.The patients’DFS (Disease-Free survival)and OS (overall survival)without FLT3 expression were 65% and 86% in 24 months’following investigation,but the patients’ DFS and OS with FLT3 expression were 20% and 40% respectively.The difference was distinct between them(P<0.05). Conclusion FLT3 can be thought as a new marker for worse prognosis of AML patients,and its expression level is associat-ed with maglignacy-grade.

Objective: To observe the influence of transcutaneous CO2emission (TCE) at points on the meridian or beside the meridian after different stimulations like acupuncture De-qi, sham acupuncture, study the correlation between acupuncture De-qi and local energy metabolism through TCE. Methods: Twenty healthy volunteers (male 8, female 12, average age 29 years old) were selected. Before, during and after five different acupuncture Neignan (PC 6), the TCE was measured at Quze (PC 3), point A [at the lower one fourth of line between Tianquan (PC 2) and Quze (PC 3)], point B [at the middle of line between Ximen (PC 4) and Quze (PC 3)], point C (2 cm ulnar side point A) and point D (2 cm ulnar side point B) with a high sensitive C02 instrument (Frewil-Q.F., made in China). The measure results were compared. Results: The comparison on value of TCE before and after the acupuncture, in Sham acupuncture and shallow acupuncture groups, it increased in all five points; in De-qi acupuncture group, there are significant increase on point A, point B, and Quze (PC 3), it has statistic significance, but there are no significant increase on point C and D. After Ximen (PC 4) or Daling (PC 7) were pressed to block Neiguan (PC 6), the value of TCE on Quze (PC 3), point A and point B were decreased during acupuncture Neiguan (PC 6), and increased after the block was removed, but there was no significant difference. Conclusion: Sham acupuncture has also some effects on body, it can increase energy metabolism of local skin, it may be the reason that shallow acupuncture without qi amving can obtain clinical efficacy. The operation method of traditional qi arriving acupuncture can increase energy metabolism of relevant meridian skin, mechanical press can reduce this effect.

Objective To investigate the phenotype and function of CD8+T cells cultured with SK-OV-3. Methods Transwell coculture experiments were conducted in 24-well plates with inner wells to separate CD8+ T cells and SK-OV-3. After 5 days of culture, CD8+ T cells were washed, and 1 × 106 cells were collected for Foxp3, CD25, CD28, CTLA-4 and GITR mRNA analysis and 2 × 106 cells were collected to detect expression of Foxp3, CD25, CD28, CTLA-4 and GITR in CD8+ T cells by flow cytometry. CD8+T cells that cultured alone or with SK-OV-3 were added at ratios of 1∶0 , 1∶1 , 1∶5 , 1∶10 , and 0∶1 to na?ve CD4+ T cells in 96-well plates. All wells were cultured with the presence of irradiated PBMCs and anti-CD3 antibody. After 72 h, [3H]-thymidine was added for 16 h prior to the determination of proliferation by scintillation counting. Results Compared with CD8+ T cells cultured without SK-OV-3 , the expression of Foxp3 and CTLA-4 was increased and CD28 expression was decreased in CD8+ T cells cultured with SK-OV-3 (both P < 0.001). We found that CD8+T cells cultured with SK-OV-3 significantly suppressed the na?ve CD4+ T cell proliferation induced by the anti-CD3 stimulation in a dose-dependent manner. In contrast, CD8+ T cells cultured without SK-OV-3 did not suppress na?ve CD4+ T cell proliferation. Conclusion Ovarian cancer cell can induce the suppressive CD8+Treg, which is an important link of the immunosuppressive microenvironment in ovarian cancer.

Objective: In the authors' previous study, acupuncture had an effect along meridians. The effect ofacupuncture can be represented by local energy metabolism which can be expressed by transcutaneous CO2 emission (TCE). To study the transportation of the effect of acupuncture and the influence to the transportation when blocking the meridians, the following experiment was carried out. Methods: Twenty healthy volunteers (male 8, female 12, average age 29) were examined on pericardium meridian at points between Quze (PC 3) and Tianquan (PC 2) (Point a), between Ximen (PC 4) and Quze (PC 3) (Point b), two control points (c and d) beside a and b and Quze (PC 3). TCE was measured on these five points before, during and after acupuncturing Neiguan (PC 6) by a high sensitive CO2 instrument (Frewil-Q.F., made in China). Mechanical pressure of 1 kg (2 cm～ 1 cm area) was given on Ximen (PC 4) or Daling (PC 7) during the acupuncture and TCE were measured before acupuncture, during acupuncture and mechanical pressing, withdrawing acupuncture but keeping pressing and after withdrawing the pressure. Results: TCE increased significantly on point a, b and Quze (PC 3) during and after acupuncture without mechanical press. When mechanical press was given at Ximen (PC 4), no significantly increase was found on above three points during and after acupuncture. When pressing Daling (PC 7), there was still significant (P<0.05) increase at Quz～ (PC 3) and point during acupuncture and significant increase (P<0.05) at point b after acupuncuture while pressing was kept. Conclusion: Acupuncture can enhance the energy metabolism expressed by TCE along meridians and this effect can be eliminated by mechanical pressing along meridians on the way between acupuncture point and measured point.

ObjectiveTo explore the effect of combined usage of expanding coronal decompressive craniectomy and sequential dural incision for treating severe bilateral frontal contusion (SBFC).Methods Forty-three patients with SBFC were randomly divided by sequential single day after hospitalization into two groups.Observation group(23 cases) treated with expanding coronal decompressive craniectomy and sequential dural incision.Control group (20 cases) treated with standard hemicraniectomy and routine dural incision.ResultsThe occurrence rate of acute cephalocele was significantly lower in observation group [ 17.4％(4/23) ] than that in control group [ 55.0％(11/20) ] (P ＜ 0.05).According to Glasgow outcome scale (GOS) score of six-month observation after operation,17 cases(73.9％,17/23) of observation group got favourable recovery or moderate deficit,other 6 cases(26.1％,6/23) got severe deficit,persistent vegetative status or death.While only 8 cases(40.0％,8/20) got favourable recovery or moderate deficit,12 cases ( 60.0％,12/20 ) got severe deficit,persistent vegetative status or death in control group.The rate of favourable recovery and moderate deficit of two groups had statistical significance (P ＜ 0.05).ConclusionCombination application of expanding coronal decompressive craniectomy and sequential dural incision is an effective method to treat patients with SBFC,and can obviously improve the rate of successful rescue and decrease the rate of mortality and disability.

Objective To assess the patient enablement level in a general hospital and its related factors,to investigate the relationship of patient enablement with patient's overall satisfaction and doctor's empathy levels.Methods Patients attending the Family Medicine Clinic of HKU-Shenzhen Hospital in September 2014 and met the inclusion criteria were asked to complete a questionnaire survey which included general characteristics,overall satisfaction;the Patient Enablement Instrument (PEI) and the Consultation and Relational Empathy Measure Scale (CARE) after the consultation.Results Total 2 326 questionnaires were issued and 1 818 were retrieved,of which 1 478 were qualified questionnaires.The qualified rate of PEI was 81.3％ and the qualified rate of patient overall satisfaction was 96.28％.The positive rate of patient enablement was 97.70％ (1 444/1 478),and the mean score of PEI was 8.21 ±3.33.Multivariate linear regression analysis showed that the PEI scores were not influenced by gender and education level of the patients,gender of the consulting doctors and initial/follow up consultation.The PEI scores were influenced by the age of the patients,the types of clinic (general practice/chronic practice clinic) and the reasons for consulting (P ＜ 0.05).There were correlations between PEI score and overall satisfaction of patients (r =0.383,P ＜ 0.001),patient's recommendation of the clinic (r =0.595,P ＜ 0.001) and CARE scores (r =0.546,P ＜ 0.001).Conclusion The results show that the PEI scores of patients visiting family medicine clinic of this hospital are higher than those reported in other studies.The PEI score may be influenced by the age of the patients,the types of clinic and the reasons for consulting.Positive correlations between patient enablement and the overall satisfaction,patient's recommendation and doctor empathy were observed.

ObjectiveTo investigate the gene expressions of TAZ and Wnt/β-catenin on the postosteogenic cells of mesenchymal stem cells(MSC)in multiple myeloma(MM)patients and to explore the potential therapeutic target of multiple myeloma bone disease (MBD).MethodsBone marrow mononuclear cells MNC from MM and controls were isolated,cultured,expanded and then induced to osteogenic differentiation.Realtime quantitative RT-PCR was employed to detect the osteogenic markers (TAZ,Wnt/β-catenin,OPN,OC,ALP and Cbf α1); and alizarin red staining for mineral deposition.The mRNA expressions of TAZ and Wnt/β-catenin in the two groups were analysed.ResultsAlizarin red staining was positive and the red calcium nodules were appeared on the post-osteogenic cells of MSC.The mRNA expressions of OC,ALP and Cbf α1 were 2.0958±0.5665,2.6670±0.3847,0.8463±0.3473,respectively,on the post-osteogenic cells of MSC in the experimental groups,which were significantly higher than those of pre-osteogenic cells(1.3487±0.9291,1.1452±0.6054,0.4439±0.2945) (t =2.171,6.709,2.795; all P ＜ 0.05).The mRNA expressions of OPN,OC,ALP and Cbf α1 were 2.1096±0.8267,2.8991±0.3531,4.3045±0.2844,1.3273±0.4075,respectively,on the post-osteogenic cells of MSC in the controls,which were significantly higher than those of pre-osteogenic cells (1.2200±0.9091,0.8780±0.3927,1.9161±0.2684,0.6736±0.2513) (t =2.289,12.103,25.134,4.411; all P ＜ 0.05).The mRNA expressions of OPN,OC,ALP,Cbf α1 were 1.2710±0.5636,2.0958±0.5665,2.6670± 0.3847,0.8463+0.3473,respectively,on the post-osteogenic cells of MSC in the experimental groups,which were significantly lower than those of control groups(2.1096 ±0.8267,2.8991 ±0.3531,4.3045±0.2844,1.3273±0.4075) (t =-2.650,-3.805,-10.822,-2.841; all P ＜ 0.05).The mRNA expression of TAZ and β-catenin were 2.2315±1.0723 and 0.5801±0.2159 on the post-osteogenic cells of MSC in MM patients,which were significantly lower than those of control groups (4.4140±0.8325,0.9516±0.2920) (t =±5.085,-3.235;both P ＜ 0.05).ConclusionThe gene expressions of OPN,OC,ALP and Cbf α1,the osteogenesis related genes,are increased in post-osteogenic cells of MSC,which showed the MSC have been successfully induced to osteoblasts.Comparing with control groups,the osteogenic potential of MSC in MM patients is lower.Based on the above research,TAZ and Wnt/β-catenin may present a novel target for the future therapy of MBD.

Objective To detect the expression of heat shock protein 47(HSP47) in renal proximal epithelial cell lines (HK-2) and to investigate the role of HSP47 in the progress of transforming growth factor β1 (TGF-β1) induced epithelial-mesenchymal transdifferentiation (EMT) in HK-2 cells.Methods HK-2 cells were exposed to TGF-β1 (0,2.5,5,10 μg/L) for different time (0,12,24,48 h).The expression of HSP47 was examined by Western blotting.Then HK-2 cells were exposed to 10 μg/L TGF-β1,the expressions of vimentin,zona occludens-1 (ZO-1) were examined by Western blotting and real-time PCR.Furthermore,the expressions of p-Smad3 and Smad3 were examined by Western blotting.HK-2 cells were transfected with HSP47 siRNA and siRNA negative control before exposing to TGF-β1.Then the expressions of vimentin,ZO-1 were detected by Western blotting and real-time PCR,meanwhile Western blotting for HSP47,p-Smad3 and Smad3.Results Stimulating HK-2 with TGF-β1 resulted in a significant increased expression of HSP47 in time-and concentration-dependent manner (P ＜ 0.05).Meanwhile,TGF-β1 up-regulated the protein and mRNA expression of vimentin (P ＜ 0.05),and down-regulated the protein and mRNA expression of ZO-1 (P ＜ 0.05),all in time-dependent manner.Stimulating HK-2 with TGF-β1 resulted in phosphorylation of Smad3,which was peaked at 30 min,slightly decreased at 1 h,and then increased again between 24 and 48 h (P ＜ 0.05).Compared to the TGF-β1 group,inhibition of HSP4.7 expression in HK-2up-regulated the protein and mRNA expression of ZO-1,down-regulated the protein and mRNA expression of vimentin (P ＜ 0.05) and down-regulated the ratio of p-Smad3/Smad3.HSP47 siRNA negative control had no significant effect on the expressions of ZO-1,vimentin and p-Smad3/Smad3 (P ＞ 0.05).Conclusion HSP47 can promote the EMT of renal tubular epithelial cell which is possibly via the TGF-β1-Smad3 pathway.