BACKGROUND:As a good candidate for bioresorbable bone graft, carbonate apatite monolith can be prepared by sintering procedure;however, sintering can cause carbonate loss and result in a much lower rate of
biodegradation compared to the human bone, thereby influencing the formation of new bone.
OBJECTIVE:To fabricate low-crystal inity carbonate apatite monolith as bone graft and test its properties. METHODS:Calcium carbonate monolith prepared by carbonation of Ca(OH) 2 monolith was treated in 1 mol/L
RESULTS AND CONCLUSION:The calcium carbonate completely transformed into low-crystal inity B-type
carbonate apatite after treated for 14 days. Diametral tensile strength of the final product was (10.27±1.08) MPa, which is adequate as a reconstruction material for bone defect. The carbonate content was (4.80±0.50)%, similar to that of the nature bone. The molar Ca/P was 1.63±0.01, indicating the Ca-deficient carbonate apatite is
obtained. The present method al ows an easy fabrication of low-crystal inity B-type carbonate apatite block with adequate strength and without sintering process.
ammonium dihydrogen phosphate solution at 60 ℃ up to 14 days. Diametral tensile strength was examined for biomechanical properties;X-ray diffraction analysis, Fourier transform infrared spectroscopy, scanning electron microscope observation and chemical analyses (carbonate, calcium and phosphate content) were also performed for physical and chemical properties.

With the application of monoclonal antibody technology more and more widely, its production technology is becoming more and more perfect. Small molecule monoclonal antibody technology is becoming a hot research topic for people. The application of traditional Chinese medicine small molecule monoclonal antibody technology has been more and more widely, the technology for effective Chinese medicine component knockout provide strong technical support. The preparation of monoclonal antibodies and small molecule knockout technology are reviewed in this paper. The preparation of several steps, such as: in the process of preparation of antigen, hapten carrier coupling, coupling ratio determination and identification of artificial antigen and establishment of animal immunization and hybridoma cell lines of monoclonal antibody, the large-scale preparation; small molecule monoclonal antibody on Immune in affinity chromatography column method is discussed in detail. The author believes that this technology will make the traditional Chinese medicine research on a higher level, and improve the level of internationalization of Chinese medicine research.
Animals ; Antibodies, Monoclonal ; chemistry ; genetics ; immunology ; Humans ; Hybridomas ; metabolism ; Immunologic Techniques ; methods ; trends

Objective To enrich pancreatic cancer stem cells through culturing mammospheres, and to detect the expressions of epidermal growth factor receptor(EGFR) and a disintegrin and metalloprotease 9 (ADAM9) and investigate their significance. Methods PANC1 cells were cultured in serum-free conditioned medium to continuously generate mammospheres, and parts of mammospheres were cultured on a collagen substratum to induce differentiation. Mammospheres cells and differentiated cells were collected, flow cytometry was used to detect the proportion of side population (SP) cells, and the expressions of EGFR, ADAM9 mRNA and protein were detected by real-time PCR and Western blotting. Results PANC1 cells mammospheres were successfully generated and could be passed continuously. After differentiation, mammospheres cells could regain the ability of adherent growth. The proportion SP cells in mammospheres cells and differentiated cells were ( 5.40 ± 0.38 ) % and (2.80 ± 0.42 ) %, and the difference was statistically significant ( P ＜ 0. 05 ).Compared with differentiated cells, the expression of EGFR and ADAM9 mRNA of mammospheres cells upregulated 2.5 and 3.0 folds ( P ＜ 0. 05 ). The expressions of EGFR and ADAM9 protein of mammospheres cells were 0.90 ± 0. 09 and 0.64 ± 0.07, which were significantly higher than those in differentiated cells (0.62 ±0.11 and 0.48 ±0.09, P ＜0.05). Conclusions Mammosphere cells contained higher proportion of pancreatic cancer stem cells. ADAM 9 may play an important role in the occurrence and development of pancreatic cancer through the EGFR signaling pathway.

In order to investigate the effect of malic enzyme on anaerobic mixed acid fermentation,the gene encoding NAD+-dependent malic enzyme was amplified from E.coli DH5? genome by using PCR and cloned into vector pTrc99a to give an expression vector pTrc99a-sfcA.Malic enzyme was overproduced by E.coli FMJ39(ldh,pfl)harboring pTrc99a-sfcA under the condition of IPTG induction.The results of anaerobic fermentation indicated overexpression of malic enzyme would have influence on the pathway of formate,acetate and succinate of FMJ39.The concentration of formate and acetate was 17.58% and 15.27% higher than FMJ39,the succinate concentration was reduced by 26.87%.There was no obvious citrate concentration change.The experimental results also indicated that high concentration of L-Thr and L-Ser would induce the Tdc operon to convert pyruvate into formate and acetate even when pfl gene encoding the pyruvate formate lyase was inactivated.

Animals ; Dentate Gyrus ; Hippocampus ; Rats ; Receptors, Adrenergic, beta ; Sleep ; Spatial Learning

OBJECTIVETo observe the levels of high mobility group box-1 protein (HMGB1), tumor necrosis factor-alpha (TNF-α), IL-6, troponin I (Tn I) release in septic rats, and to explore themechanism of Taohong Qinlian Decoction (TQD) in the treatment of septic myocardial injury.

METHODSA total of 48 healthy male Wistar rats of clean grade were randomly divided into the sham-operation group (Sham), the sepsis model group (CLP), and the TQD treatment group (ZY), 16 in each group. Concen-trations of TNF-α, IL-6, Tn I, and HMGB1 expression were detected in each group at 24 and 48 h after operation. Pathological changes of cardiac muscle were observed under light microscope.

RESULTSConcentrations of TNF-α, IL-6, Tn I and HMGB1 at 24 and 48 h after operation were significantly higher in the CLP group than in the Sham group (P < 0.01). Concentrations of TNF-α, IL-6, Tn I, and HMGB1 at 24 and 48 h after operation were significantly lower in the ZY group than in the CLP group (P < 0.05). Myocardial injury occurred in the CLP and the ZY group under light microscope. And this injury was more severe in the CLP group than in the ZY group.

CONCLUSIONTQL could reduce the level of sepsis-related inflammatory cytokines and protect myocardium in septic rats.

Animals ; Drugs, Chinese Herbal ; pharmacology ; HMGB1 Protein ; metabolism ; Heart ; drug effects ; Interleukin-6 ; metabolism ; Male ; Myocardium ; metabolism ; pathology ; Random Allocation ; Rats ; Rats, Wistar ; Sepsis ; pathology ; Troponin I ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective To establish a pre -column derivatization and HPLC method for the determination of free amino acids in serum and urine with phenyl isothiocyanate ( PITC ) as derivatization reagent . Methods The determination was performed on waters e 2695 sys-tem.Sample was separated by Shim -pack VP-ODS column(250 mm × 4.6 mm,5 μm ) with a gradient elution of mobile phases composed of methanol/acetate/H2 O( pH 6.5 ) , the content of amino acid were tested by UV at 254 nm.Results The developed method had a good linear ( r>0.991 8 ) in the content range of 2-200 μg· mL-1 , with an aver-age recovery ranged from 88.5% to 123.4%.The precision was within 0.12%-3.42%.Conclusion PITC pre-column derivatization and HPLC method was developed and validated , which had good accuracy , suitable for the determination amino acid in serum and urine of patients with diabetes .

Objective: To explore the patterns of Syndrome Differentiation (SD) of coronary heart disease (CHD) patients in peri-operative stage of coronary artery bypass graft (CABG). Methods: One week after operation, thirty-seven CHD patients, who received CABG of internal mammary artery or great saphena vein under conventional general anesthesia with low or middle temperature extracorporeal circulation were differentiated as various syndromes, with the pre- or post-operational EKG, color Doppler echocardiography were done during and after operation. The hemodynamic parameters were monitored. Results: In the CHD patients, 64.9% were differentiated as Qi-Yin deficiency, 67.6% were complicated with phlegm syndrome and 62.2% with blood stasis, suggesting that Qi-deficiency, phlegm and stasis are the basic pathogenetic factors in patients with CABG. Moreover, the peri-operative syndrome was correlated with the condition of coronary artery lesion, heart and lung functions before operation, and the extracorporeal circulation time during the operation. Conclusion: TCM SD conducting in peri-operative stage might be useful in exploring the patterns of syndrome alteration which provided a basis for preventing peri-operative complications and elevating success rate of operation.

Animals ; Fatty Liver, Alcoholic ; drug therapy ; Female ; Male ; Phytotherapy ; Rats ; Rats, Wistar ; Saponins ; therapeutic use ; Trigonella

BACKGROUND:Idiopathic interstitial pneumonia is of poor response to treatment. Glucocorticoids are the first medicine for the treatment, however there is only 30% of the patients who are responded. Traditional Chinese drugs (TCD) have been researched hot point for prevention and treatment of pulmonary fibrosis. Many TCD have been used clinically, and with a certain therapeutic effect. Transforming growth factor-β1 and tumor necrosis factor-α are the considerable cytokines to cause pulmonary fibrosis, inhibition of their expression, therefore, may be effective to pulmonary fibrosis.OBJECTIVE: To investigate the interventional effect of qidan granule on pulmonary fibrosis in rats induced by bleomycin A5 and the influence on the expressions of transforming growth factor-β1 and tumor necrosis factorα, and also to compare with those of hydrocortisone.DESIGN:A randomized and interval grouping design.SETTING:Department of Respiratory Medicine, Shandong Provincial Hospital, Shandong University.MATERIALS:The experiment was conducted from May 2003 to March 2004 at Pathological Laboratory of Shandong Provincial Academy of Medical Science. Totally 105 SD male rats, were at random divided into 4groups: normal control group (n= 15 ), model group, qidan group and hydrocortisone group, with 30 rats in each group. Each group was subdivided as7-day, 14-day and 28-day group, with 5 rats in each normal group, and 10in each other groups.METHODS: [1] Model establishment: A perfusion was intrabronchially performed, of 0.25 mL normal saline for rats in normal control group, and of bleomycin A5 0.25 mL ( 5 mg/kg,4 g/L) for rats in other 3 groups, to set up the models of pulmonary fibrosis. [2]Administration: Next day to the beginning of modeling qidan granule (consisting of Radix Astragali seu Hedysari, Radix Salviae Miltiorrhizae, Rhizoma Ligustici Chuanxiong and so on, 3 125 mg/kg) was intragastrically given per day for rats in qidan group, hydrocortisone (25 mg/kg) was intraperitoneally given per day for rats in hydrocortisone group, and normal saline (2 mL/rat) was intragastrically given per day for rats in normal and model groups.[3] Observation indexes: The rats in each group were on the day 7, day 14 and day 28 put to death under the anesthesia, then the lung tissue was taken, stained with hematoxyline-eosin stain for pathological observation of lung tissue. The expressions of transforming growth factor-β1 and tumor necrosis factor-α were detected by immunohistochemistry.MAIN OUTCOME MEASURES:Pathological observation of lung tissue,and the expressions of transforming growth factor-β1 and tumor necrosis factor-α at different time points of rats in each group.RESULTS:Totally 100 rats entered the final result analysis.[1]Pathological observation of lung tissue: In the normal group the structure was normal, in the model group there were alveolitis on the day 7, deterioration of alveolitis on the day 14, and extensive fibrosis on the day 28; the degrees of alveolitis and fibrosis in the qidan group were slighter than those in the model group, and there was normal structure of alveoli; and in the hydrocortisone group the alveolitis on the day 7 and 14 was slighter than that in the model group, but there was no significant difference of fibrosis compared with the model group.[2] Expression of transforming growth factor-β1:In the model group the expression was highest on the day 28 and obviously higher than that in the normal group (3.6±0.4,1.2±0.4,P ＜ 0.01 ); the expression in the qidan group and hydrocortisone group was obviously lower than that in the model group(1.7±0.5,2.5±0.4,P ＜ 0.01), and the expression in the qidan group was lower than that in the hydrocortisone group (P＜ 0.01 ). [3]Expression of tumor necrosis factor-α: In the model group at different time points the expression was continuously increased, the expression in the qidan group and hydrocortisone group was obviously lower than that in the model group(P ＜ 0.05 or P ＜ 0.01), and the expression in the qidan group was lower than that in the hydrocortisone group ( P ＜ 0.01).CONCLUSION: Qidan granule can obviously reduce the extent of pulmonary fibrosis in rats induced by bleomycin A5, lower the expressions of transforming growth factor-β1 and tumor necrosis factor-α, and the effect was better than that of hydrocortisone.