Objective: To study the preparation technology of glycyrrhetinic acid (GA)-tanshinone IIA (Tan IIA) compound liposomes (GT-Lip). Methods: The compound liposomes were made of soybean phosphatidylcholine (SPC) and cholesterol (Ch) by film dispersion ultrasonic probe method. Hydration temperature and ultrasonic power were optimized by single factor tests and the optimum formulation was selected by orthogonal design. The encapsulation efficiencies (EE) of GA and Tan IIA were determined by low-speed centrifugation. The particle size and Zeta potential of liposomes were detected by dynamic light scattering particle size meter. Transmission electron microscopy was used to observe morphous. Results: The optimal preparation conditions were as follows weight ratio of SPC-Ch 6:1, molar ratio of SPC-Tan IIA and SPC-GA was 30:1 and 24:1, hydration temperature 30 °C, and ultrasonic power 380 W for 5 min. Using the optimal method, the EE values of GA and Tan IIA in GT-Lip were (81.50 ± 0.76)% and (98.63 ± 0.90)% (n = 3), and the average particle size of liposomes was (120.5 ± 1.62) nm and the Zeta potential of liposomes was (-19.00 ± 0.98) mV (n = 3). Conclusion: The optimal preparation method of GT-Lip in this study is stable and feasible.

Objective To approach the expressions and the roles of connective tissue growth factor (CTGF) and membrane type 1 matrix metalloproteinases (MT1-MMP) in valve disease with pressure overload which induces extracellular matrix remodeling of left ventricle. MethodsOf 32 patients, 16 cases were pressure overload group (PO), who had the multiple valve disease with predominately aortic valve stenosis, having a ring diameter of aorta valve less than 1.3 cm, cross valve pressure gradient equal or more than 40 mmHg, and valvular regurgitation less than 4.0 cm2; The other 16 cases, as mitral stenosis group (MS), were simple mitral stenosis patients with single valve replacement. Meanwhile, 5 normal individuals served as control, who died from accident. Echocardiography was used to analyze the left ventricular function and detect the hypertrophic level of the left ventricle. Left ventricle muscle samples were obtained during operation. Histological features were studied by Masson staining, and collagenous contents were quantitated with a computer-assisted imaging analysis system. The mRNA expressions of CTGF and MT1-MMP were detected with RT-PCR. ResultsConcentric hypertrophy was observed significant in PO group, but myocardial hypertrophy was not found in MS group. Compare to the MS group and control, PO group had significantly more collagenous contents in left ventricle, thickened vessel wall, and narrow lumen of blood vessel (P0.05), but CTGF mRNA expression was increased in MS group compare to control (P

Human tumor necrosis factor-related apoptosis-inducing ligand (hTRAIL) might be developed as a novel anti-tumor drug due to its selective cytotoxicity in tumor cells. The predicted Macaca mulatta TRAIL (mmTRAIL) is highly homologous to hTRAIL in nucleotide acid as well as amino acid sequence, suggesting that mmTRAIL might induce apoptosis of human cancer cells. However, the cytotoxicity of mmTRAIL in human cancer cells has not been investigated. In this paper, it is reported that the gene encoding mmTRAIL has been cloned by using reverse-transcriptase polymerase chain reaction (RT-PCR) from monkey peripheral blood mononuclear cells (PBMCs) in our laboratory. Subsequently, an expression plasmid was constructed by inserting mmTRAIL gene into pQE30 plasmid. After induction by addition of Isopropyl β-D-1-Thiogalactopyranoside (IPTG), mmTRAIL was expressed. MmTRAIL was recovered from supernatant of sonicated bacteria by Ni-NTA agarose affinity chromatography. SDS-PAGE and gel filtration chromatography demonstrated that mmTRAIL forms trimer in solution. In vitro assays indicated that mmTRAIL was cytotoxic to human COLO205 tumor cells but not to normal cells at low concentration of nanomole. In addition, antitumor effect of mmTRAIL was evaluated in mice bearing human COLO205 tumor xenografts. Intratumorally injected mmTRAIL significantly inhibited growth of tumor grafts. These results suggested that mmTRAIL was valuable as candidate drug for cancer-targeted therapy.
Animals ; Antineoplastic Agents ; Apoptosis ; Cell Line, Tumor ; Cloning, Molecular ; Humans ; Leukocytes, Mononuclear ; Macaca mulatta ; Mice ; Plasmids ; TNF-Related Apoptosis-Inducing Ligand ; genetics ; metabolism ; Xenograft Model Antitumor Assays

Objective To investigate the possible effect of citrate on electrolyte metabolism in healthy people with different genders and races and provide a reference for the possible clinical interventions.Methods A cross over,placebo-controlled study was conducted in 22 age-matched Chinese(11 males and 11 females)and 10 male Caucasian volunteers after informed consents were obtained.Volunteers received of saline solution,separated by a wash-out period of two to three weeks.Serial blood and urine samples were collected during the observation period and analyzed for the selective biochemical parameters.Results Comparable basal levels of serum albumin[male(43.05±1.81)g/L vs female(42.26±2.67)g/L]and serum ionized calcium[male(1.27±0.04)mmol/L vs female(1.26±0.04)mmol/L]were observed between different genders of Chinese volunteers.However,citrate intervention led to more pronounced decrease of ionized calcium level in Chinese females compared to Chinese males[-28.68%(-20.00%--35.2%)vs-23.84%(-16.53%--29.32%),t=3.19,P ＜ 0.01].There was no differences of the levels of serum inorganic phosphate[-18.81%(-3.16%--25.09%)vs-19.23%(-1.22%--32.16%),t=0.36,P＞0.05]and albumin[-0.32%(3.27%--7.60%)vs 1.88%(6.03%--9.31%),t=0.47,P＞0.05].Independent of gender,citrate intervention resulted in an increased excretion of urine calcium in Chinese volunteers[before 0.34(0.09-0.87)vs after 0.96(0.18-1.47),t=6.66,P ＜0.01].Compared to Caucasian males,Chinese males has a higher basal level of serum ionized calcium [(1.27±0.04)mmol/L vs(1.22±0.02)mmol/L,t=3.7,P ＜0.01]and larger amplitude basal rhythm in serum albumin level[-11.72%(-5.70%--14.21%)vs-1.74%(2.43%--7.68%),t=7.43,P ＜ 0.01].Application of citrate resulted in comparable changes of serum ionized calcium [-23.84%(-16.53%--29.32%)vs-21.95%(-18.31%--30.92%)],phosphate[-19.23%(4.65%--32.16%)vs-12.68%(0.68%--42.19%)],albumin[-0.32%(1.05%--7.60%)vs-1.39%(1.87%--7.26%)]and urine calcium excretion[237.70%(11.8%-935%)vs 234.37%(5.45%-504.00%)]between Chinese and Caucasian males(t=0.32,0.03,0.25 and 0.04 respectively,P＞0.05).Serum levels of magnesium were not influenced in all volunteers during two interventions.Conclusions Independent of race and gender,the invention of citrate results in comparable changes of serum magnesium,inorganic phosphate and albumin.The effect of citrate on ionized calcium levels between genders implicates a higher risk for hypocalcemic reactions in females compared to males undergoing automatic apheresis procedures.

AIM:To study the expression level of S100 calcium-binding protein A4 (S100A4) in synovial tissue of the knee joint in rheumatoid arthritis (RA) patients and normal persons, and the effect of S100A4 on the angiogenesis induced by rheumatoid arthritis fibroblast-like synoviocytes (RAFLSs).METHODS:The synovial tissue was taken from the knee joint of the RA patients (RA group) and the normal persons (control group).The protein expression of S100A4 and vascular endothelial growth factor (VEGF) in the synovial tissue of the 2 groups was observed by immunohistochemistry.RAFLSs were isolated from synovial tissue of patients with active RA.ELISA was used to detect the effect of S100A4 on the secretion of VEGF by RAFLSs.The effect of S100A4 on the angiogenesis of HUVECs cultured with conditioned medium from RAFLSs was also detected.RESULTS:The protein of S100A4 and VEGF was highly expressed in the synovial tissues of RA group (P<0.05).rhS100A4 significantly stimulated the secretion of VEGF in RAFLSs in a time-and dose-dependent manner (P<0.05).Cultured with conditioned medium from RAFLSs, rhS100A4 significantly promoted HUVECs to form tube-like structures in vitro.CONCLUSION:S100A4 protein is highly expressed in synovial tissue of the knee joint in RA patients, and S100A4 stimulates synovial angiogenesis by promoting RAFLSs to generate VEGF, indicating that S100A4 may be used as a potential target for the treatment of RA.

AIM To prepare the moisture-proof granules of polysaccharides from Astragali Radix.METHODS Mixture design was adopted in the optimization of excipient proportion after moisture absorption determination of mixed powder of polysaccharides and seven excipients.With the influencing factors of ethanol concentration,ethanol amount,drying temperature and drying time,together with the evaluation indices of moisture absorption rate and non-forming rate,the preparation was optimized by orthogonal test based on single factor test.The critical relative humidities were then compared after drawing moisture absorption curves of polysaccharides,mixed powder and granules.RESULTS The mixed powder with an optimal ratio (1 ∶ 2) of polysaccharides to mixed excipients (46.3％ lactose,14.5％ mannitol and 39.2％ microcrystalline cellulose) was found to remain the moisture absorption rate of 11.6％ within 168 h.The optimal conditions were determined to be 70％ for ethanol concentration,0.125 times for ethanol amount,55 ℃ for drying temperature,and 60 min for drying time,the moisture absorption rate was 8.1％ within 168 h,and the non-forming rate was 11.46％.Compared with polysaccharides and mixed powder,the granules showed relatively lower initial velocity and acceleration (absolute value) of moisture absorption,and relatively higher critical relative humidity.CONCLUSION The granules of polysaccharides from Astragali Radix prepared by this method show a good moisture-proof effect.

Objective　To explore the injurious effect of the serum drawn from patient under cardiopulmonary bypass (CPB) on pulmonary surfacant-associated protein A(SP-A) and the preventive effect of pentoxifylline (PTX) on the protein. Methods　The cultured rat alveolar epithelial cell type Ⅱ(AT-Ⅱ) were incubated with CPB serum to observe the change of the cell morphology, and the expressions of SP-A and SP-A mRNA. Results　Traumatic changes and the decrease of SP-A and SP-A mRNA expressions were found in AT-Ⅱ cells cultured with CBP serum. PTX exerted protective effect on the cells. Conclusion　The serum after CPB can directly injure rat AT-Ⅱ cells in vitro, and inhibit the SP-A expression at transcription and translation levels, which probably is an important reason for the quantitative and qualitative abnormality of pulmonary surfactant after operation. PTX may alleviate the inhibitory effect of CPB serum on SP-A.

OBJECTIVE: To investigate the in vivo possibility of osteogenesis and angiogenesis of tissue-engineered periosteum in rabbits.METHODS: The marrow mesenchymal stem cells (MSCs) derived from New Zealand rabbits were adhered to small intestinal submucosa (SIS) to fabricate the tissue-engineered periosteum. Totally 12 New Zealand rabbits were received critical bone defect in bilateral radii to prepare models. The tissue-engineered periosteum was randomly implanted in one side of bone defect,and the other side was treated by SIS. At 4 weeks after operation, the angiogenesis of tissue engineered bone was detected by Tetracycline fluorescence microscopy and formaldehyde-ink perfusion method; simultaneously, the new bone formation was firmed by haematoxylin-eosin staining.RESULTS: Animals showed normal daily behaviors and non-infection wounds healing. The gross observation showed that bone defects in the experimental side were bridged with newly formed bone; while the defects of the control side were remained empty.Tetracycline fluorescence microscopy and hisotological examination could confirm the new bone tissue formation in the experimental side. The ink staining in new bone specimens suggested that there were abundant of neovasculization in tissue-engineered bone.CONCLUSION: Tissue-engineered periosteum can form new bone in allogenic rabbits and can be vascularized by some inherent mechanism for new bone tissue survivor.

Health Personnel ; psychology ; Humans ; Logistic Models ; Multivariate Analysis ; Professional Competence ; statistics & numerical data ; Stress, Psychological ; epidemiology

Acrocephalosyndactylia ; diagnosis ; genetics ; therapy ; Female ; Humans ; Infant