Azure Stains ; chemistry ; DNA, Neoplasm ; analysis ; chemistry ; Humans ; Rosaniline Dyes ; chemistry ; Staining and Labeling ; economics ; methods ; Stomach Neoplasms ; genetics ; pathology

Objective To explore the detailed underlying molecular and signaling mechanisms in the effects of icariin on bone formation by an in vitro cell model. Methods The proliferation of MC3T3-E1 osteoblast-like cells was evaluated by MTT, and gene expression of cell cycle related proteins in MC3T3-E1 cells after icariin treatment was detected by real-time PCR. The phosphorylation of MAPK signals, including ERK, P38, and JNK was determined by Western blot, and then the inhibitors of MAPK signals were used to treat cells with icariin alone or together to determine the role of MAPKs in the process of icariin treatment on MC3T3-E1 cell proliferation. Alkaline phosphatase and Alizarin red staining were used to detect the formation of mineralization nodules, and gene expressions of alkaline phosphatase, type Ⅰ collagen, and osteocalcin in osteoblasts after being treated by icariin were evaluated by real-time PCR. ICI182780, and nilutamide was used to decide the participation of estrogen and androgen receptor signals in the process of icariin treatment on the differentiation and mineralization of MC3T3-E1 cells. Results Treatment with icariin promoted MC3T3-E1 cell growth in a time- and dose-dependent manner. This treatment also revealed that icariin increased the expression of mRNAs encoding both cyclin E and PCNA, positive regulators of cell growth, but decreased levels of mRNAs encoding Cdkn2b, a negative regulator of cell cycle progression. When MC3T3-E1 cells were cultured in a differentiated condition, icariin enhanced mineralized nodule formation and increased the expression of mRNAs encoding alkaline phosphatase, type Ⅰ collagen, and osteocalcin. Treatment with icariin significantly induced phosphorylation of both ERK and JNK and this phosphorylated effect occurred very rapidly within 5 minutes and reached peak at 15 minutes. Furthermore, the stimulated effects of icariin on proliferation and gene expression of cyclin E, PCNA, and Cdkn2b in MC3T3-E1 cells were dramatically attenuated by treatment with both U0126 and SP600125, inhibitors of MAPKs. Interestingly, such stimulating effects of icariin were at least partly reduced by treatment with ICI182780, an inhibitor of estrogen receptor. Icariin induced mineralized nodule formation and gene expression of alkaline phosphatase, type Ⅰ collagen, and osteocalcin in MC3T3-E1 cells were also partly reduced when the cells were treated with ICI182780. Conclusions Our findings indicate that the anabolic effect of icariin on bone formation is, at least partly, mediated through the MAPK signaling pathway in order to modulate osteoblast proliferation and differentiation.

Objective To assess the coordinated regulation and the molecular mechanisms of 17β-estradiol and 1,25-dihydroxyvitamin D3 [ 1,25-( OH ) 2 D3 ] on the proliferation and differentiation of osteoblastic MC3T3-E1 cells.Methods MC3T3-E1 cells were cultured in phenol-red free α-MEM medium supplemented with 10％ FBS,MTT assay was performed to determine the effects of 17β-estradiol and 1,25-( OH )2 D3 on MC3T3-E1 cells proliferation.After cells were treated with different agents,cell cycle related genes [ cyclin E,proliferation cell nuclear antigen ( PCNA ),and cyclin-dependent kinase inhibitor 2b ( Cdkn2b ) ] and markers of osteoblastic differentiation [ type Ⅰ collagen ( COL Ⅰ ),alkaline phosphatase ( ALP),osteopontin ( OPN ) ] were detected with SYBR green-based quantitative PCR.ALP activity was detected with BCIP/NBT method.Results 17β-estradiol could promote proliferation of MC3T3-E1 cells,which was accordant to its ability to increase cyclin E and PCNA and to inhibit Cdkn2b mRNA expression in MC3T3-E1 cells.However,1,25-( OH)2D3 had no effect on the proliferation of MC3T3-E1 cells and also did not enhance the proliferation of MC3T3-E1 cells stimulated by 17β-estradiol.On the other hand,17β-estradiol promoted the gene expression of differentiation markers Col Ⅰ,ALP,and OPN,and 1,25-(OH) 2 D3 synergistically increased the expression of these genes with 17 β-estradiol.Conclusion As two of the most important hormones which regulate bone metabolism,estrogen and vitamin D may coordinately promote osteoblast differentiation,but may not regulate osteoblasts proliferation synergistically.

In order to identify Peucedani Radix, Peucedani Decursivi Radix and their adulterants, the internal transcribed spacer 2 (ITS2) regions of Peucedani Radix, Peucedani Decursivi Radix and their adulterants were amplified and bidirectionally sequenced based on the Principles for Molecular Identification of Traditional Chinese Materia Medica Using DNA Barcoding, which has been promulgated by Chinese Pharmacopoeia Commission. Sequences were analyzed and assembled by Codon Code Aligner V3. 7.1. The relevant data were analyzed by MEGA 5. 0. Species identification analyses were performed by using the nearest distance methods and neighbor-joining (NJ) methods. The result showed that the ITS2 sequence lengths of Peucedani Radix were 229-230 bp and the average intra-specific genetic distances were 0.005. The ITS2 sequence lengths of Peucedani Decursivi Radix were 227 bp and the sequences contained no variation site. The average inter-specific K2P genetic distance of Peucedani Radix, Peucedani Decursivi Radix and their adulterants species were 0.044 and 0.065 respectively. The minimum inter-specific divergence is larger than the maximum intra-specific divergence of Peucedani Decursivi Radix. The nearest distance methods and NJ trees results indicated that Peucedani Radix, Peucedani Decursivi Radix and their adulterants species could be identification clearly. The ITS2 regions can stably and accurately distinguish Peucedani Radix, Peucedani Decursivi Radix and their adulterants.
Apiaceae ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Ribosomal Spacer ; Drug Contamination

The average life expectancy in Macau is ranking the second in the world, the consumption of fresh medicinal plants is a profound culture in Macau. The paper focus on the distribution of the antioxidant herbs, a comprehensive investigation and analysis the amount of the plant resources was carried out. The antioxidant activity of alcohol extracts was determined by using the DPPH method, and six kinds fresh herbs with high antioxidant free radical activity were screened out. Reference to adult daily dose of vitamin C, it is calculated that the daily dose amount of fresh herbs is less than 200 g. For the expected shortage of resources and the ecological status of Macau, we give some suggestions of herbal introduction in population ecology reconstruction.
Antioxidants ; pharmacology ; Ecosystem ; Free Radical Scavengers ; pharmacology ; Macau ; Plant Extracts ; pharmacology

Objective To evaluate the effectiveness of clinical rotation training in pain department for general practitioner.Methods Totally 31 general practitioners of 3 years program in the First Affiliated Hospital of Chongqing Medical University were involved in clinical rotation training in pain department form August 2008 to December 2011.Training lasted for one month.Clinical rotation training in pain department was conducted for general practitioners by studying rules and regulations,treatment methods,imaging data and clinical practice.Evaluation was made according to the treatment,theoretical knowledge,interpersonal skill and professional competency of general practitioners.Measurement data were expressed as mean ± standard deviation ((-x) ± s) while enumeration data were expressed as rate.Descriptive analysis of the data was used.Results Average total number of patients managed by 31 general practitioners was (15 ± 2),average total number of disease species was (5 ±2),average number of case discussion and small lecture participated was 1 ～2,average number of nerve block participated as assistant was (40 ± 5) and average number of minimally invasive surgery visited and studied was (2 ± 2).Residency duties were skillfully completed and assessment score was (85 ± 4.50).Conclusions Through clinical rotation training,general practitioners can understand common disease diagnosis and treatment category; familiarize clinical evaluation methods and master standards of diagnosis and treatment in pain department.Meanwhile,they would learn doctor-patient communication to establish a good therapeutic relationship with patients.

Objective To investigate the effects of icarrin on the activity and protein expression of core binding factor otl(Cbfa1) in rat osteoblasts cultured in vitro,and to explore whether mitogen-activated protein kinase (MAPK) pathway is involved in this process.Methods Calvarial osteoblasts were obtained from newborn (<24 h) SD rats by trypsin-coUagenase digestion method.The second generation osteoblasts were cultured in the medium containing icariin (10 ng/ml) or estradiol (10-8 mol/L) with or without extracellular-signal regulated kinase (ERK) inhibitor (UO126) or p38MAPK inhibitor (SB203580).Nuclear protein was extracted from osteoblasts.And then the activity of Cbfa1 was detected by ELISA.The amounts of Cbfa1 protein were detected by Western blot.Results Calvarial osteoblasts were obtained successfully and were used in this study after indentified by alkaline phosphatase and mineralized nodus staining.Cbfa1 expression and the activity in osteoblasts were up-regulated by both icariin and estradiol (P<0.05).The effects were partly inhibited by addition of U0126or SB203580 (P<0.05).Conclusions Either icarrin or estradiol can stimulate the proliferation and maturation of cultured osteoblasts in vitro via up-regulating the activity and expression of Cbfal.The MAPK signal pathway inhibitor seems to partly decrease Cbfa1 activity.It suggests that MAPK pathway may be involved in the transduction of icariin's impact on proliferation and mineralization of osteoblasts.

Objective To investigate BAALC(brain and acute leukemia cytoplasmic)gene expression in patients with de novo acute myeloid leukemia(AML)and its clinical significance. Methods BAALC expression was determined by real-time quantitative polymerase chain reaction(RQ-PCR) in 63 de novo AML patients.The association between BAALC expression and therapeutic effect was analyzed.Results The correlation coefficiencies were over 0.99 for standard curves of RQ-PCR method. BAALC expression was detected in 49(78%)AML patients.The peripheral WBC counts,hemoglobin, platelet counts and the bone mahow blast cell percentage at onset in 31 AML patients with high BAALC expression were(26.3?18.1)?10~9/L,(78.3?21.8)g/L,(76.9?64.5)?10~9/L and(61.2?22.3)% and those of 32 AML patients with low BAALC expression were(30.2?21.7)?10~9/L,(81.6?30.9)g/L, (73.9?57.2)?10~9/L,(54.3?16.3)%,respectively.No statistic differences were found between these two groups.The AML patients with normal chromosome karyotypes are more likely to have a high BAALC expression(68%)compared with those with abnormal chromosome karyotypes(23%,?~2=12.093,P= 0.001).AML patients with normal cytogenetics and high BAALC expression shows significant lower CR rate (65%)compared with those with low BAALC expression(84%,?~2=6.573,P=0.013). Conclusion High BAALC expression may define an important risk factor in AML with normal cytogenetics and predicts an adverse prognosis.

Forty type 2 diabetic patients with diabetic peripheral neuropathy (DPN) were assigned to two groups and treated respectively with?-lipoic acid or mecobalamin for 2 weeks.The results suggested that?-lipoic acid could accelerate the nerve conduction velocity and decrease the plasma level of endothelin and C reactive protein as well as microalbuminceria with a effect similar to mecobalamin therapy on DPN.

Intraoperative cell salvage ( IOCS) was conducted during cesarean delivery from May 2011 to April 2016 in our hospital: when the volume of autologous blood collected from the surgical field≥800 ml during surgery and the parturients′hemoglobin≤100 g∕L after hemorrhage, the autologous blood was centrifuged, concentrated, washed and then filtered using a leucocyte depletion filter before reinfusion. IOCS was carried out in 1 085 cases during cesarean delivery within 5 yr. For the parturients who underwent IOCS, the percentage of patients who did not require transfusion of allogeneic red blood cells was 82.95%(900 cases), the percentage of patients who required plasma transfusion was 41.66% (452 cases), the percentage of patients who required platelet transfusion was 3.32% (36 cases), and no autologous blood transfusion?related adverse reactions were found during surgery and hospital stay. In conclusion, IOCS could be safely and effectively used for blood?saving effect in cesarean delivery.