Artesunate Decreases ß-Catenin Expression, Cell Proliferation and Apoptosis Resistance in the MG-63 Human Osteosarcoma Cell Line.

BACKGROUND: This study aims to determine the effects of artesunate on proliferation, apoptosis and ß-catenin expression in the human osteosarcoma cell line MG-63. METHODS: MG-63 cells in the logarithmic growth phase were collected and cultured with different concentrations of artesunate (12.5 µg/mL, 25 µg/mL and 50 µg/mL) for 24 h, 48 h and 72 h. The total number of MG-63 cells and the morphological changes were observed under an inverted microscope. The MTT assay was adopted to test the inhibition rate (IR) of cell growth. The apoptosis rate was detected using annexin V/propidium iodide (PI) staining. Cell cycle distribution was identified by flow cytometry (FCM), and the expression levels of ß-catenin, cyclins and cyclin dependent kinases (CDKs) were measured using Western blotting. RESULTS: The results of the MTT assay indicated that artesunate could remarkably inhibit MG-63 cell proliferation compared with the rates in the untreated control group (0 µg/mL artesunate), and the inhibitory effect was dose-dependent. The apoptosis rate of MG-63 cells was elevated as the concentration of artesunate increased, and all the rates were significantly higher than that in the control group. Additionally, as the artesunate concentration increased, the proportion of MG-63 cells in G0/G1 phase gradually declined whereas that of cells in the G2/M and S phases increased. Western blotting confirmed that a higher concentration of artesunate reduced the expression levels of ß-catenin, cyclin A, cyclin D1 and CDK1 and increased the expression levels of cyclin B1; however, artesunate had no impact on CDK2 expression in MG-63 cells. CONCLUSION: These results demonstrated that artesunate can inhibit ß-catenin expression and cell proliferation as well as promote cell apoptosis in MG-63 cells, which indicates that artesunate may serve as a promising drug in the clinical treatment of osteosarcoma.

GIT1 gene deletion delays chondrocyte differentiation and healing of tibial plateau fracture through suppressing proliferation and apoptosis of chondrocyte.

BACKGROUND: Although tibial plateau fracture is an uncommon injury, its regulation is challenging and there are some influencing factors, including the effects of severe bone displacement, depression and cancellous bone cartilage, and inevitable cartilage damage. And GIT1 plays an important role in bone mass and 78 osteoblast cell migration. METHODS: The study used 72 C57/BL6 mice. A tibial plateau fracture model was established by using mice with the same number of GIT1 gene deletions (the experimental group) and their wild-type littermates (the control group). Joint and bone callus recovery were evaluated by X-ray and CT thin layer scans. Micro CT assay and histomorphometry were conducted in order to evaluate the volume of newly formed blood vessels. Type II collagen expression in tibial tissues after tibial plateau fracture were detected by immunohistochemistry after 7, 14 and 21 days. The number of proliferating cell nuclear antigen (PCNA) positive cells after tibial plateau fracture was tested by immunohistochemistry after 14 and 21 days. The terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining was conducted after 14 and 21 days in order to test chondrocyte apoptosis in tibial tissues after tibial plateau fracture. RESULTS: The GIT1 gene deletion group mice spent less time on the rotating rod than the control group mice (P < 0.05). Compared with the control group, postoperative recovery was retarded, because GIT1 gene deletion slowed down neovascularization after tibial plateau fracture (P < 0.05). Compared with the control group, mouse type II collagen expression significantly decreased in the GIT1 gene deletion group, and the proportion of PCNA positive cells significantly decreased (P < 0.05). The TUNEL results indicate that GIT1 gene deletion led to reduced chondrocyte apoptosis. CONCLUSION: GIT1 gene deletion can inhibit chondrocyte proliferation and apoptosis during the recovery of tibial plateau fracture, so as to delay chondrocyte differentiation and tibial plateau fracture healing.

Bridging external fixation versus non-bridging external fixation for unstable distal radius fractures: A systematic review and meta-analysis.

OBJECTIVE: A systematic review and meta-analysis was conducted to compare the relative effectiveness of bridging external fixation and non-bridging external fixation for distal radius fractures treatment. METHOD: Relevant literature were comprehensively searched using the PubMed, Springer Link, Karger Medical and Scientific Publishers, Chinese Biomedical Database (CBM) and Chinese National Knowledge Infrastructure (CNKI) databases without any language restrictions. STATA Version 12.0 software and Comprehensive Meta-analysis 2.0 were applied. RESULTS: A total of 905 patients with distal radius fracture from six eligible cohort studies were selected for statistical analysis. Our meta-analysis results indicate that the non-bridging cases had a higher risk of pin track infection, rupture of the extensor pollicis longus and nerve injury than the bridging cases. Subgroup analysis stratified by country indicated non-bridging patients showed evidence of an increased risk of pin track infection and higher risk of rupture of the extensor pollicis longus compared with the patients treated with bridging external fixation in the UK population. The follow-up results showed flexion degree of patients treated with non-bridging external fixation was slightly better than that of patients treated with bridging external fixation (P < 0.05). CONCLUSION: There is evidence in our systematic review and meta-analysis to support that bridging external fixation can reduce the incidence of pin tract infections and nerve injury compared to non-bridging external fixation, but have no significant difference in other complications and the recovery of wrist joint function. Bridging external fixation could therefore be a better choice in patients with distal radius fractures.

[Distribution laws of Chinese medical syndrome types and analyses of risk factors in senile hypertension patients: a clinical study].

OBJECTIVE: To explore the distribution laws of TCM syndrome types and to analyze the distribution of dynamic blood pressure curve, atherosclerosis, and age in senile hypertension patients. METHODS: Totally 1 131 senile hypertension patients were recruited from 7 provinces and municipal cities. Features of TCM syndromes, classification and distribution curves, and syndrome distribution laws were observed. The distribution curves of dynamic blood pressure, carotid atherosclerosis, and age were compared in each TCM syndrome types. RESULTS: There were four main syndrome types in 736 cases (56.15%), i.e., excessive accumulation of phlegm-dampness syndrome (210 cases, 16.02%), yin deficiency and hyperactivity of yang syndrome (177 cases, 13.50%), Gan-Shen yin deficiency syndrome (79 cases, 6.03%), and deficiency of qi and yin syndrome (252 cases, 19.22%). Besides, there were two more sub-types, i.e., collateral obstruction by blood stasis syndrome and collateral obstruction by phlegm and stasis. Circadian blood pressure monitor was completed in 211 cases. Of them, abnormal circadian blood pressure occurred in 152 cases (accounting for 72. 38%); yin deficiency and hyperactivity of yang syndrome, excessive accumulation of phlegm-dampness syndrome, deficiency of qi and yin syndrome plus collateral obstruction by blood stasis syndrome were most often seen. Color ultrasound of carotid artery was performed in 660 patients of main syndromes. The incidence was quite higher in those of excessive accumulation of phlegm-dampness syndrome (182 cases, 27. 58%), deficiency of qi and yin syndrome plus collateral obstruction by blood stasis syndrome or collateral obstruction by phlegm and stasis (322 cases, 48.79%). Yin deficiency and hyperactivity of yang syndrome was dominant in patients 60 -79 years old, while deficiency of qi and yin syndrome and Gan-Shen yin deficiency syndrome were dominant in patients older than 80 years. CONCLUSIONS: Excessive accumulation of phlegm-dampness syndrome, yin deficiency and hyperactivity of yang syndrome, Gan-Shen yin deficiency syndrome, and deficiency of qi and yin syndrome were main syndrome types in senile hypertension patients. There was statistical difference in the distribution curves of blood pressure, atherosclerosis, and age of various TCM syndrome types.

The distribution of IGF2 and IMP3 in osteosarcoma and its relationship with angiogenesis.

The aim of this study was to investigate the expression patterns of IGF2 and IMP3 in osteosarcoma as well as its relationship with angiogenesis in the tumor. IGF2 and IMP3 expression was detected by immunohistochemical staining in the serial sections of the osteosarcoma. The impacts of IGF2 and IMP3 expression patterns on tumor angiogenesis were evaluated by statistics. The IGF2 and IMP3 staining had different expression patterns in different osteosarcoma. Twelve out of the sixty-four cases of conventional osteosarcoma showed nuclear staining patterns, and twenty-nine showed cytoplasmic staining of IGF2 and IMP3 simultaneously. On the other hand, fourteen cases showed nuclear IGF2 staining but cytoplasmic IMP3 expression, and nine cases showed nuclear IMP3 staining and cytoplasmic IGF2 expression. Twenty-eight out of forty-seven cases of parosteal osteosarcoma showed nuclear IGF2 and IMP3 expression, nine showed cytoplasmic IGF2 and IMP3 expression simultaneously. Seven out of forty-seven cases of parosteal osteosarcoma expressed IGF2 with nuclear staining but expressed IMP3 with cytoplasmic staining. Meanwhile, three cases expressed IGF2 with cytoplasmic staining but expressed IMP3 with nuclear staining. Similar to the parosteal osteosarcoma, the periosteal osteosarcoma expressed IGF2 and IMP3 mainly with nuclear staining simultaneously, forty out of fifty-five cases of periosteal osteosarcoma did that. Five out of fifty-five cases expressed IGF2 and IMP3 with cytoplasmic staining at the same time. Four cases showed nuclear IGF2 staining and cytoplasmic IMP3 staining. In the parosteal and periosteal osteosarcoma, there was no significant difference in IGF and IMP3 expression patterns (P = 0.216). However, compared with conventional osteosarcoma, the parosteal and periosteal osteosarcoma showed significant difference in IMP3 and IGF2 expression (P = 0.016, P = 0.023). IGF2 and IMP3 expression patterns were positive correlation in the different osteosarcoma (r = 0.1021, P = 0.032). The Microvessel density (MVD) in osteosarcoma with IGF2 and IMP3 cytoplasmic staining was more than that with nuclear expression of IGF2 and IMP3, and the difference was significant (P = 0.024). Moreover, the conventional osteosarcoma with cytoplasmic IGF and IMP3 showed more MVD than parosteal and periosteal osteosarcoma with cytoplasmic IGF and IMP3, and the difference was significant (P = 0.035). IGF2 and IMP3 had different expression patterns, which might be associated with angiogenesis. However, cytoplasmic and nuclear expression of IGF2 and IMP3 might play different roles in the angiogenesis of osteosarcoma.

[Distribution characteristics of syndrome types in essential hypertension].

OBJECTIVE: To investigate the distribution characteristics of syndrome types of traditional Chinese medicine (TCM) in essential hypertension and to explore the distribution rule of TCM syndromes. METHODS: A multicenter, large-sample survey method of clinical epidemiology was applied to choose the patients with essential hypertension from North, Middle, and South China. A questionnaire was designed and filled in, then 477 untreated patients with first-diagnosed essential hypertension were selected and the information was recorded into FileMaker database. A cluster analysis method was utilized to study the TCM syndrome distribution rule of essential hypertension. RESULTS: Two-step cluster analysis was done from 3 to 7 clusters. Seven clusters were appropriate, which included deficiency of heart and kidney qi, hyperactivity of liver-yang, deficiency of yin and yang, stagnation of phlegm-dampness, phlegm-heat (subtype of stagnation of phlegm-dampness), blood stasis obstructing collaterals, and other syndromes. The symptoms presenting high percentage in each cluster were more significant in TCM theory. The syndromes of hyperactivity of liver-yang (24.1%) and stagnation of phlegm-dampness (27.1%) presented the high percentages, and deficiency of heart and kidney qi (10.1%), deficiency of yin and yang (8.4%), and blood stasis obstructing collaterals (9.0%) presented the low percentages. CONCLUSION: As compared with the current syndrome differentiation criteria, two-step cluster analysis results not only include the syndromes of deficiency of yin and yang, hyperactivity of liver-yang, stagnation of phlegm-dampness, but also cover qi deficiency and blood stasis.

[Potential of using pattern diagnosis of traditional Chinese medicine to improve the clinical use of antihypertensive agents].

Despite the availability of six classes of antihypertensive agents, control of blood pressure and improving patients' quality of life remain far from ideal. There is a wide variability in terms of the hypotensive effect and side effect profile for the same antihypertensive agent used in different patients. How to select the right agent to provide the most beneficial results in terms of efficacy and improvement of quality of life as well as to decrease clinical symptoms and minimize adverse reactions is an important therapeutic challenge. It has been suggested that clinical usage of pattern (Zheng) diagnosis of traditional Chinese medicine may improve the accuracy in selecting the right antihypertensive agents with improved efficacy and deceased adverse effects. Limited research in this area suggested the calcium channel blocker may work better in treating phlegmatic damp excess pattern and blood stasis pattern while beta-blockers may be more beneficial in the liver yang rising pattern. On the other hand, angiotensin converting enzyme inhibitors may be more suitable in a yin deficiency and yang hyperactivity pattern as well as combined liver and kidney yin deficiency pattern. More research studies using this innovative approach in improving the selection of antihypertensive agents including mechanistic studies are urgently needed.

[Exploration of TCM syndrome differentiation of coronary heart disease and coronary arteriography].

Coronary arteriography (CAG) examination is the widely accepted gold standard for diagnosis of coronary heart disease (CHD) nowadays. In order to explore the situation and value of CAG applied in TCM syndrome differentiation of CHD, and to facilitate the research on disease-syndrome diagnosis of CHD in integrated traditional Chinese and Western medicine (ICWM), the relationship between coronary arterial pathological changes and the TCM syndrome types was probed through reviewing literatures concerning the application of CAG in studying syndrome differentiation of CHD since 2000. The results showed that syndrome types are correlated to the severity and number affected of coronary artery branches. Along with the aggravation of CHD, TCM syndrome typing become even more complicated. There is a gap between the previous detectable index for CHD as well as the TCM syndrome typing and clinical practice. CAG is of vital importance in exploring rules of integrated syndrome differentiation and disease diagnosis of CHD. Therefore, to launch a nationwide multi-centric study on large sample of syndrome differentiation with ICWM is necessary, which should be based on the evidence-based medicine and by dint of the modern medical detecting technique to conduct the study comprehensively in combining differentiation of syndrome and disease, and in both macroscopic and microscopic views.

[Gaseous signaling molecule and study on activating blood circulation and removing stasis].

The finding of gaseous signaling molecule NO, CO and H2S, and the advocacy of "gas biology" concept have provided a novel thinking for the study of Huoxue Huayu (HXHY, a traditional Chinese medicine method of promoting blood circulation to remove blood stasis). The concept "Qi" in TCM and gaseous signaling molecule are correlated in respects of their source and mesomeric function. Study of HXHY could only be deepened through an overall cognition on blood stasis syndrome with the view of Qi-blood correlation.