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Objective: To develop a method for the determination of lacidipine (LAC) in human plasma.Methods: After liquid-liquid extraction with tert-butyl methyl ether, the plasma samples were analyzed by LC-MS/MS.Using lacidipine-13C8 as the internal standard, a Agilent ZORBAX Eclipse XDB C18 column (150 mm×2.1 mm, 5 μm) was used with the mobile phase consisting of water(containing 5 mmol·L-1 ammonium formate)-acetonitrile(15∶85,v/v)at a flow rate of 0.3 ml·min-1 and with the column temperature at 40 ℃.The ion transitions were performed in a positive electrospray ionization multiple reaction-monitoring mode regarding + as the molecular ion peak of lacidipine and monitoring with m/z 473.5→m/z 410.3, m/z 473.5→m/z 400.1 and m/z 473.5→m/z 354.3.The internal standard was monitored with m/z 481.4→m/z 362.3.Results: The linear range of lacidipine was 0.1-10 ng·ml-1 (r＞0.99) and the lower quantification limit was 0.1 ng·ml-1.The intra-and inter-day RSDs were 3.15%-7.04% and the relative error was from-8.58% to 12.71%.The mean relative recovery of lacidipine was from 107% to 118% (RSD＜15%).The plasma samples were stable at-20℃ for 40 d and kept stable after three repeated freeze-thaw cycles.The prepared samples were stable at room temperature for 24 h and in the automatic sample injector (4℃) for 24 h(RSD＜15%).Conclusion: The developed assay method can be applied in the bioequivalence evaluation and pharmacokinetic studies of lacidipine in human.
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BACKGROUND:In recent years, many reports have focused on inflammatory cytokines, growth factors and mechanical loads affecting the cartilage and subchondral regeneration, but there is a lack of comprehensive understanding about the mechanism of osteoarthritis. OBJECTIVE: To explore the correlation between function status of bone marrow mesenchymal stem cels and disease progression in patients with osteoarthritis. METHODS:Femoral bone marrow was extracted from patients with femoral neck fractures (control group), mild (mild group) and severe (severe group) osteoarthritis to isolate and culture bone marrow mesenchymal stem cels. Cel counting kit-8 was used to detect the proliferative ability of bone marrow mesenchymal stem cels from different patient groups, and passage 3 bone marrow mesenchymal stem cels were subject to 2-week chondrogenic induction folowed by toluidine blue staining. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cels were isolated and cultured from the femoral bone marrow of different groups. The proliferative ability of cels in the control group was significantly higher than that in the mild and severe groups. After chondrogenic induction, bone marrow mesenchymal stem cels varied obviously in the morphology that was from fusiform to qusi-circular or polygon, the percentage of nucleoplasm became smaler, and cels were positive for toluidine blue staining. The number of chondrocytes generated in the severe group was less than that in the control group, but there was no great difference in cel morphology. These findings indicate that the occurrence of osteoarthritis is negatively correlated with the functional status of autologous bone marrow mesenchymal stem cels.
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BACKGROUND:Since damage control theory system was founded, this theory in the orthopedics has been applied gradualy, especialy in elderly hip fracture surgery that reduces the negative impacts due to inflammatory responses. OBJECTIVE:To explore whether flurbiprofen axetil can reduce inflammatory responses in rats with hip fractures based on the damage control theory. METHODS: Forty-nine healthy Sprague-Dawley rats weighing 250-300 g were randomly divided into four groups:control group (n=7), immediate internal fixation group (n=14), flurbiprofen axetil group (n=14), damage control group (n=14). Rats in the control group moved freely in the cages. Rats in the other three groups were intraperitonealy injected with composite anesthetics to make unilateral hip fracture models, and then respectively given internal fixation immediately after fracture, flurbiprofen axetil injection and delayed internal fixation, and delayed internal fixation. Levels of serum C-reactive protein, interleukin-6 and tumor necrosis factor-α were determined and analyzed before fixation, immediately after internal fixation and at 4, 8, 12, 24, 48 hours after internal fixation in different groups. RESULTS AND CONCLUSION:Postoperative serum levels of C-reactive protein, interleukin-6, tumor necrosis factor-αwere al increased in different groups. The level of C-reactive protein reached the peak at 24 hours after internal fixation. Flurbiprofen axetil injection had no significant influence on the level of C-reactive protein in rats with delayed internal fixation (P=0.51). Interleukin-6 levels were stil increased at 48 hours after internal fixation, but flurbiprofen axetil reduced the level of interleukin-6 significantly in rats with delayed internal fixation (P < 0.01). The tumor necrosis factor-α level peaked at 4 hours after internal fixation, and flurbiprofen axetil injection could significantly reduce the level of tumor necrosis factor-α in rats with delayed internal fixation (P < 0.01). These findings indicate that flurbiprofen axetil as a new non-steroidal anti-inflammatory drug can reduce the inflammatory response in rats with hip fractures after internal fixation, and also can aleviate the inflammatory response of rats undergoing delayed operation under the guidance of damage control theory.
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BACKGROUND:Bone marrow mesenchymal stem cels are crucial for bone and cartilage development and regeneration at a celular level. Insufficient quantity and functional impairment of bone marrow mesenchymal stem cels is widely considered to be one of osteoarthritis causes. OBJECTIVE: To explore the relationship between the functional status of bone marrow mesenchymal stem cels and disease progression in osteoarthritis patients.METHODS: Thirty patients with osteoarthritis were enroled from July 2013 to October 2014, and divided into control, mild osteoarthritis, and severe osteoarthritis groups, with 10 cases in each group. 5 mL bone marrow from the femur or tibia was extracted from each patient to isolate and culture bone marrow mesenchymal stem cels. Proliferation ability of cels at passage 3 was detected using cel counting kit-8; toluidine blue staining was performed at 14 days after chondrogenic induction; real-time PCR was used to detect the mRNA expression of Aggrecan and Col2A1 in the control group after chondrogenic induction. RESULTS AND CONCLUSION:Afterin vitro culture, bone marrow mesenchymal stem cels grew adherently in polygonal and fusiform shape with multiple processes at uniform size. The cytoplasm contained larger particles and the nuclei were ovoid. Most of cels were in cel division phase. The proliferation ability was strongest in the control group and weakest in the severe osteoarthritis group. Cels from the three groups were al at plateau phase after 1 week culture. At 14 days after chondrogenic induction, the cels were polygonal and quasi-circular, and purple metachromatic granules distributed outside of the cytoplasm. The expression of Aggrecan and Col2A1 in the control group displayed an overexpression trend. These findings indicate that the functional status of bone marrow mesenchymal stem cels from osteoarthritis patients is negatively correlated with the severity of disease, which can influence the disease progression in osteoarthritis patients.
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BACKGROUND:At present, the anatomic reconstruction of coracoclavicular ligament is a trend for the treatment of acromioclavicular joint dislocation. Endobutton plate is a new and effective surgical method found in recent years. Although the recent curative effect is satisfactory, as a method of internal fixation, its rigid fixation does not conform to the requirements of the biomechanics. Furthermore, a randomized control ed clinical study and systematic evaluation are absent. OBJECTIVE:To systematical y evaluate clinical outcomes and safety between endobutton plate and the other surgical methods for treatment of acromioclavicular joint dislocation. METHODCochrane (2014-01), PubMed (1966-01/2014-03), Medline (1966-01/2014-03), EMbase (1984-01/2014-03), CNKI (1979-01/2014-03), VIP (1989-01/2014-03) and Wanfang (1989-01/2014-03) databases were searched by computer. Six Chinese journals about orthopedics were searched by hand. References of relevant literatures were searched. Randomized control ed trials that were related to different surgical methods for the treatment of acromioclavicular joint dislocation were col ected. In accordance with inclusion criteria, some literatures were included and their qualities were assessed strictly. Meta-analysis was performed with RevMan 5.2 software from the Cochrane Col aboration. RESULTS AND CONCLUSION:Final y 7 published studies with randomized control ed trials met al the inclusion criteria. A total of 359 patients were included, containing 152 cases of endobutton plates fixation, 149 cases of clavicle hook plate fixation, and 58 cases of screw fixation. Except two researches addressed the comparison among three surgical ways, the remaining were about two ways. Meta-analysis showed that the excellent rate was better in endobutton plates fixation than in clavicle hook plate fixation (P=0.0002) and in screw fixation (P=0.009). Compared with other fixation methods, there were significant differences in shoulder joint pain (P=0.01), but no significant difference was detected in operation time, upper limb muscle strength, bleeding amount, redislocation and surgical wound infection (P=0.44, P=0.10, P=0.37, P=0.21, P=0.96). Results indicated that, the efficiency of endobutton plates fixation for acromioclavicular joint dislocation was better than clavicle hook plate fixation and screw fixation, and caused less shoulder pain than clavicle hook plate fixation. There was no significant difference in operation time, upper limb muscle strength, bleeding amount, redislocation and surgical wound infection among the three methods. Due to the limited number of cases in this study, the multicenter, large-sample and long-term clinical randomized control ed studies are needed to increase the strength of the evidence.
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BACKGROUND:At present, Rockwood type I, II acute dislocation of acromioclavicular joint is recommended to be treated non-operatively, while acute dislocation of type IV, V and VI should be treated operatively. The treatment of type III acute dislocation of acromioclavicular joint remains controversial. Furthermore, there are a variety of methods to repair the dislocation of acromioclavicular joint. Among them, the anatomical reconstruction of coracoclavicular ligament and acromioclavicular ligament to restore the biomechanics of acromioclavicular joint are the developing trend. Therefore, more and more scholars pay attention to the anatomical structure of acromioclavicular joint. OBJECTIVE:To summarize the research status of the repair and reconstruction of acromioclavicular joint dislocation. METHODS:A computer-based retrieval was performed among Wanfang database (http://g.wanfangdata.com.cn/) and PubMed database (http://www.ncbi.nlm.nih.gov/PubMed) by the first author until May 2014, for articles about acromioclavicular joint structure and acromioclavicular joint dislocation. The key words were“acromioclavicular, acromioclavicular joint dislocation, endobutton”in English, and“acromioclavicular joint dislocation, anatomical reconstruction”in Chinese. After repeated studies were excluded, 1 632 articles were screened out, and 43 of them were involved in the analysis. RESULTS AND CONCLUSION:Surgical methods of acromioclavicular joint dislocation are various. Now the treatment trends to anatomical reconstruction, including tendon transplantion and endobutton technique. Based on it, more and more basic researches of the acromioclavicular joint emerge. But due to the lack of large sample, multi-center clinical control study, the optimal treatment of the acromioclavicular joint dislocation is at the exploration stage.