Objective: To establish a RP HPLC method for determination of geniposide in Longdanxiegan Oral Liquid. Methods: A Nova Pak C 18 column was used. The mobile phase was methanol water(20:80). The detection wavelength was 240nm. Results: The linear range of geniposide was 0.569～1.707?g, r=0.9998. The average recovery was 98.67%, RSD=1.54% (n=6). Conclusion: This method is convenient, rapid and accurate. It can be used for quality control in production of Longdanxiegan Oral Liquid.

Objective To identify the effect of long hazardous drinking on cardiovascular function and cardiovascu?lar abnormalities among alcohol dependent patients. Methods A follow-up survey was conducted, 72 potential patients who were diagnosed as having alcohol dependence were recruited into case group and 75 staff who underwent routine health examination were subjected into control group. Furthermore, 52 patients were subdivided into long hazardous drinking group (GroupⅠ) according to the classification of alcohol consumption published by WHO. The rest patients in the case group were considered as not long hazardous drinkers (GroupⅡ). The blood lipid data, echocardiography and ca?rotid artery brachial artery ultrasonography measurement data were compared between the three groups. The high risk fac?tors for cardiovascular abnormalities among alcohol dependence patients were analyzed. And one year after discharge, telephone follow-up method was used to obtain the incidence of cardiovascular accident among patients. Results The dis?tribution of blood lipid data among GroupⅠ, Ⅱ and control group were not significantly different (P>0.05). The LVEF score in GroupⅠwas significantly lower than that in control group (P<0.01). The LAAEF score in GroupⅠwas signifi? cantly higher than that in control group and that in the GroupⅡ(P<0.05). While the FDM and IMT score in the GroupⅠwas significantly lower than that in control group (P<0.01). In the case group, the duration of drinking alcohol was neg?atively associated with LAPEF (r=-0.246, P=0.014) and LAAEF (r=0.239, P=0.016). The average daily alcohol consump?tion was positively associated with LVEF (r=0.256, P=0.010), while negatively correlated with FMD (r=-0.256,P=0.010). Multivariate logistic regression analysis showed that long hazardous drinking was an independent risk factor for cardiovas?cular abnormalities (OR=1.334, 95%CI: 1.060~1.678). Conclusion Long hazardous drinking can reduce left ventricular diastolic and vascular endothelial function. It is an independent risk factor for cardiovascular abnormalities in alcohol de?pendent patient.

OBJECTIVE:To study the inhibitory effects of phosphatidylinositol 3-kinase (PI3K) inhibitor LY29400 (“LY”) combined with indole-3-carbinol (I3C) on the proliferation of human anaplastic thyroid carcinoma FRO cells. METHODS:RFO cells in logarithmic growth phase were divided into blank control group,I3C (250 μmol/L) group,LY (10 μmol/L) group and combination (I3C of 250 μmol/L + LY of 10 μmol/L) group,which were subject to 24,48 and 72 h drug action. MTT method was used to determine and calculate the inhibitory rates of the cells in all groups,flow cytometry assay to determine the apoptosis rates after 48 h action,Western blot method to determine the expressions of Capspase-3 proteins thereafter and immunohistochemi-cal method to determine the expressions of Bcl-2 and Bax thereafter. RESULTS:The inhibitory rate obviously increased with the ex-tension of drug action time,with statistical significance at each time point in combination group (P<0.05). Compared to I3C group and LY group,combination group had higher inhibitory rate,apoptosis rate and the expressions of Capspase-3 protein and Bax,and lower Bcl-2 expression and ratio of Bcl-2 to Bax,with statistical significance (P<0.01). CONCLUSIONS:LY com-bined with I3C has synergistic inhibitory effects on the proliferation of FRO cells by a mechanism which may be related to apopto-sis induced by down-regulating Bcl-2 expression,up-regulating Bax expression and activating Caspase cascade reaction.

BACKGROUND:Low-temperature rapid prototyping technology is a new kind of rapid prototyping technology, and it is rapidly used in the preparation of bone tissue engineering scaffolds because it can make scaffold forming control able and can keep the biological activity of the materials, also can easily realize the scaffold with porous of three-dimensional structure and other advantages.
OBJECTIVE:To investigate the preparation process of polyethylene glycol-modified polylactic acid-glycolic acid/nano-hydroxyapatite (PLGA-PEG/n-HA) using the low-temperature rapid prototyping, and to test its performance.
METHODS:PLGA-PEG/n-HA and PLGA/n-HA were prepared by low-temperature rapid prototyping equipment. Under an electron microscopy, we observed ultra-structure of the scaffolds. Immersion (ethanol) method was used to test the porosity, and electronic testing machine was used to determine the material mechanical properties. Then these two kinds of scaffolds with rat osteoblasts were cultured in vitro, the cel adhesion rate was detected by precipitation method after 12 hours, and cel counting kit-8 method was used to determine the cel proliferation at culture days 1, 3, 5, 7, 9, 12.
RESULTS AND CONCLUSION:Both of the two scaffolds had ideal aperture range and high porosity. But the aperture range of PLGA-PEG/n-HA scaffolds had large fluctuations, and the average aperture was smal er than that of PLGA/n-HA. Some pores were closed up. The cel adhesion rate and the cel growth curve of PLGA-PEG/n-HA was better than that of PLGA/n-HA (P<0.05), but the mechanical properties were less than PLGA/n-HA (P<0.05). The results showed the PLGA-PEG/n-HA scaffolds had good cel compatibility.

Objective to establish an ethical assessment system in the admittance of the limitative medical technologies.Method:literature review and Delphi method are employed to adjust and set the structure,index numbers,and index weights of ethical assessment system of the limitative medical technologies.Result:An ethical assessment system in the admittance of the limitative medical technologies was established,which included 3 primary indices,11 secondary indices and 35 tertiary indices.There were 4 qualitative levels with different weights in tertiary indices,based on which the global grade and proposed-admittance criterion were obtained.Conclusion:Establishment of ethical assessment system in the admittance of the limitative medical technologies provides an ethical evidence for the assessment of clinical admittance of the limitative medical technologies with qualitative and quantitative approaches and class-setting evidence.

BACKGROUND:The poly-lactic acid and its ramifications have many advantages, such as eligible biocompatibility,nontoxicity of degradation product, easy procession and suitable intensity. Thus, they have been widely used in bone tissue engineering.OBJECTIVE: To study the cellular biocompatibility and in vitro adhesion of poly(lactic-co-glycolic acid) (PLGA) scaffold and bone marrow stromal stem cells (BMSCs) so as to provide a basis for preparation a PLGA scaffold that can load many cytokines.DESIGN: Contrast observation.SETTING: Department of Orthopaedics and Traumatology in Nanfang Hospital of Southern Medical University.MATERIALS: The experiment was carried out in Key Laboratory of Tissue Construction and Detection of Guangdong Province from September 2004 to June 2005. One New Zealand healthy male rabbit (2 months old, 1.5-2.0 kg weight)was adopted in this study. Experimental materials: PLGA scaffold was obtained from Institute of Polymer Science in Sun Yat-sen University); beta-tricalcium phosphate (β-TCP) was supplied by AO Company (Switzerland).METHODS: Bone marrow was aspirated from the New Zealand rabbit. Mononuclear cells were harvested using whole bone marrow culturing, then were induced and amplified in the conditional culture medium. BMSCs were inoculated onto the PLGA and β-TCP at a concentration of 1 ×109 L-1, while those in the medium without materials were taken as blank control group. The development of implanted cells and the adhesion between cells and materials were observed with phase contrast microscope and scanning electron microscope. The proliferation and cycle of cells were tested with MTT method and flow cytometry.MATN OUTCOME MEASURES: ①Phase contrast microscope was used to observe the development of cells and the adherence between cells and materials at a fixed time every day. ②Cellular development on days 1, 3, 6 was observed by scanning electron microscope. ③Cellular proliferation was detected by MTT method. ④Alkaline phosphatase (ALP) activity was determined by chemical colorimetry.⑤Flow cytometer test: The effects of PLGA and β-TCP on the cellular cycle, content of DNA and polyploid levels of BMSCs were investigated. The DNA index of the candidate cells was also calculated.RESULTS: ①Phase contrast microscope observation: In the blank control group, cells culture for 7-10 days presented a larger number of shuttle-shaped, and no contact inhibition effect was observed. The time of cells beginning adherence in PLGA group was obviously later than that in β-TCP group. Cells began to develop on the circumambience and surface of the materials, with the prolonging of culture time. Most of the cells were multangular. The cells in both PLGA and β-TCP groups were similar to those in control group in items of cellular development and shape. ②Scanning electron microscope observation: On the seventh day of culture, the cells of control group remained a monolayer and amalgamated to be a patch with multangular-shaped ones increasing. There were substances in granule shape on the surface of the cells and micro-silk links between cells. In PLGA group, the cells After 7 days' cultivation proliferated sharply, and the compressed-shaped ones were inosculated to be a patch through integrations among cells, resulting in a large number of matrixes. While in β-TCP group, the number of cells increased from the 7th day. The cells were combined together to be a monolayer and moved to pores with matrixes creating out of cells.③Cellular proliferation: The number of the cells in each group all increased to some extents. However, there was no significant difference between the PLGA group and the control group, as well as the PLGA group and the β-TCP group (P ＞ 0.05).④ALP activity: The　content of ALP in all the groups enhanced without exception, while the activities between the PLGA group and the Both control group, as well as the PLGA group and the β-TCP group had no significant difference (P ＞ 0.05).⑤Cellular cycle:Both PLGA and β-TCP had slight effects on cellular cycle of BMSCs. The cells in each group were all normal diploid,and no heteroploid cells were discovered.CONCLUSION: This type of PLGA scaffold possesses good cellular biocompatibility, and can be used as a carrier of many factors in bone tissue engineering.

BACKGROUND:Nano-hydroxyapatite helps to improve the mechanical properties of bone implants.
OBJECTIVE:To study the clinical effect of nano-hydroxyapatite artificial bone on col apsed fracture of the tibial plateau.
METHODS:Fourteen cases of col apsed fracture of the tibial plateau combined with bone defects from March 2010 to September 2012 were analyzed retrospectively. The bone defect range was from 1.5 cm×1.0 cm to 3.1 cm×4.5 cm. Al patients were treated with nano-hydroxyapatite artificial bone at an implant amount of 5-14 g. Clinical and X-ray observations were applied at 1 week, 1 month and 3 months postoperatively. Hospital for Special Surgery scores were employed for recovery of knee function.
RESULTS AND CONCLUSION:The patients were fol owed up for 12-27 months. Except for one case of a smal amount of wound exudates, no general side effects occurred in 13 cases. X-ray photo showed an integrity interface between nano-hydroxyapatite artificial bone and host bone at 3 months after treatment. Primary healing was obtained in al cases without any complications. Hospital for Special Surgery score was increased to (88.7±4.3) points at 1 year later. These findings indicate that the nano-hydroxyapatite artificial bone has a good biocompatibility and biomechanics, and it may be an ideal artificial bone for repairing col apsed fractures of the tibial plateau.

BACKGROUND:Seed cells and scaffold are two key factors for cartilage defects after osteonecrosis of femoral head using tissue-engineered method. OBJECTIVE:To explore the feasibility of Bio-gide col agen membrane combined with dog bone marrow mesenchymal stem cells into chondrocytes. METHODS:Bone marrow mesenchymal stem cells were isolated from beagle dogs by whole bone marrow blood centrifugation method and adherence screening method in vitro and cultured. Morphological changes in cells were observed, and identification was done using cellsurface antigens. Bone marrow mesenchymal stem cells of passage 3 were induced by chondrocyte induction medium to differentiate into chondrocytes (experimental group). cells cultured in normal medium were considered as control group. 3-(4, 5-Dimethylthiazol-2-yl) 2, 5-diphenyl tetrazolium bromide assay was used to measure growth curve of chondrocytes. cells underwent typeⅡcol agen immunohistochemistry and toluidine blue staining. The coculture of bone marrow mesenchymal stem cells at passage 3 and Bio-gide col agen membrane were observed under an inverted phase contrast microscope and scanning electron microscope. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells with high purity and high viability were obtained by whole bone marrow blood centrifugation method and adherence screening method. cells grew wel and had strong amplified ability, and successful y differentiated into chondrocytes. Numerous bone marrow mesenchymal stem cells adhered on the Bio-gide col agen membrane, showing a tendency of multi-layer growth. cells and Bio-gide col agen membrane seem to blend into an integrant part. Cel processes appeared and connected each other and gradual y wrapped the Bio-gide col agen membrane, with the presence of obvious cel matrix secretion. These results suggested that bone marrow mesenchymal stem cells can grow and differentiate into chondrocytes on the Bio-gide col agen membrane.

BACKGROUND:There are a variety of treatments for femoral head necrosis,but their efficacy is not confirmed and unified.How to improve the differentiation ability of osteoblasts in the femoral head and improve the biomechanical support after the repair of the femoral head is an urgent problem to be solved.OBJECTIVE:To explore the clinical outcome of stem cells combined with vascularized iliac bone flap and tantalum rod implantation for the treatment of osteonecrosis of the femoral head (ONFH).METHODS:Totally 28 cases (36 hips) of non-traumatic ONFH admitted at the Zhongshan Hospital of Dalian University from January 2010 to January 2011 were enrolled.Bone marrow samples were extracted from each patient to isolate bone marrow stromal stem cells which were cultured in vitro for 2 weeks.Tantalum rod implantation with vascularized iliac bone graft was conducted to restore the femoral head shape,and then,prepared stem cell suspension were injected into the iliac bone flap and into the subchondral space of the femoral head.RESULTS AND CONCLUSION:All the 28 cases (36 hips) were followed up for 6-20 months (average 12 months),and their Harris hip scores and visual analogue scale scores at postoperative 6 and 12 months were significantly higher than the baseline (P ＜ 0.05).The Harris hip score at postoperative 12 months was significantly higher than that at postoperative 6 months (P ＜ 0.05),but there was no significant difference in the visual analogue scale scores at 6 and 12 months postoperatively (P ＞ 0.05).At the end of 12-month follow-up,clinical outcomes were excellent in 13 hips,good in 15 hips,fair in 4 hips,and poor in 4 hips,with an excellent and good rate of 90％.These findings indicate that autologous bone marrow stromal stem cell transplantation with vascularized iliac bone flap and tantalum rob implantation is an effective method with high clinical success rate for the treatment of ONFH.