The study was aimed to investigate the effect of artesunate (ART) on the apoptosis of HL-60 cells in vitro and the expression of Bcl-2 and ICAD in the process of apoptosis induced by ART. The inhibition of ART on HL-60 cells were evaluated by means of MTT assay; cell apoptosis was detected by light microscopy, agarose gel electro-phoresis, flow cytometry; Western blot was used to analyze the expression of Bcl-2 and ICAD in cells during apoptosis induced by ART. The results showed that ART could significantly inhibit the proliferation of HL-60 cells in time-and dose-dependent manner. After treating HL-60 cells with ART for 48 hours, the IC(50) values was 18.33 microg/ml and its inhibition effect contributed to the induced apoptosis. Bcl-2 and ICAD proteins both all expressed in HL-60 cells, the level of expression declined as concentration increased. It is concluded that artesunate may induce apoptosis of HL-60 cells in vitro, Bcl-2 and ICAD may be an important control factor in the signal transduction pathway of ART-induced apoptosis.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Artemisinins ; pharmacology ; HL-60 Cells ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism

To investigate me material basis of Mahuang Fuzi Xixin decoction (MFXD) for anti-inflammation and immune-suppression based on the combined method of serum chemical and serum pharmacological. The LC-MS/MS fingerprints of MFXD, drug-containing serum and blank serum were compared to define the components in plasma. Histamine, β-hexosaminidase released from RBL-2H3 cell infulenced by drug-containing serum at different time points were measured by ELISA. The effect of drug-containing serum on lipopolysaccharide-induced splenocyte proliferation at different time points were determined by MTT. A correlation analysis was made on components of MFXD and pharmacological indexes based the stepwise regression method. After the intragastrical administration with MFXD, 32 components were discovered in rat serum, including 27 prototype components (10 from Mahuang, 13 from Fuzi and four from Xixin) and five unknown components. Compared with blank serum, drug-containing serum could reduce the release of histamine from RBL-2H3 induced by antigen at different time points (P < 0.05); except the 4-hour drug-containing serum, all of the remaining drug-containing serums could inhibit the RBL-2H3 mastocyte degranulation induced by antigen at different time points (P < 0.05). Drug-containing serum could significantly lipopolysaccharide-induced mouse splenocyte proliferation at 15 and 30 min (P < 0.05). A regression analysis was made on the chemical data of components absorbed into blood and pharmacological indexes, i. e. release rate of histamine, release rate of β-hexosaminidase and inhibition rate of splenocyte. This suggested the close correlations among methyl pseudo-ephedrine, pseudoephedrine and histamine released from RBL-2H3 induced by antigen; pseudoephedrine, hypaconine, methyl pseudoephedrine and β-hexosaminidase released from RBL-2H3 induced by antigen; as well as benzoyl hypaconine, benzoylaconine, 14-benzoyl-10-OH-mesaconine, mesaconine and lipopolysaccharide-induced mouse splenocyte proliferation. Methylpseudoephedrine, pseudoephedrine, benzoyl hypaconine, benzoylaconine and mesaconine may be part of material basis of MFXD on anti-inflammation and immune suppression.
Animals ; Anti-Inflammatory Agents ; chemistry ; pharmacology ; Cell Degranulation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Histamine ; immunology ; Immunosuppressive Agents ; chemistry ; pharmacology ; Male ; Mass Spectrometry ; Mast Cells ; drug effects ; immunology ; Mice ; Rats ; Rats, Wistar ; Serum ; chemistry

AlM:To investigate the effects of pirfenidone ( PFD) on the proliferation and transfomring growth factor-β1 ( TGF-β1 ) expression in vitro culture rat corneal stromal cells.METHODS: Corneal stromal cells from 8 to 10wk SD rats were isolated, cultured and treated with different concentrations of PFD 0mg/mL (control group), 0. 15mg/mL (experimental group▏), 0. 3mg/mL (experimental group‖), 1mg/mL (experimental group Ⅲ) for 48h. CCK-8 assay was performed to assess cell proliferation, while immunocytochemistry and Western Blot were used to detect the expression of ki-67 and TGF-β1 expression, respectively. RESULTS: Compared with control group, PFD significantly inhibited the proliferation in a dose -dependent manner ( all P < 0. 05 ), so was protein expression of ki-67. PFD significantly down-regulated the expression of TGF-β1 in a dose-dependent manner (P<0. 05).CONCLUSlON: Pirfenidone can significantly inhibit the proliferation of rat corneal stromal cell by down regulating TGF-β1 expression, therefore, it has potential prospect in lightening the corneal wound healing reaction.

OBJECTIVETo evaluate the value of rectangle titanium cage (SynCage-C) in the anterior cervical spine fusion.

METHODSNineteen patients underwent anterior cervical discectomy and interbody fusion using SynCage-C with average follow-up of 9 months.

RESULTSAll cases had cervical spondylosis with 13 cases of myelopathy and 6 cases of radiculopathy. One level fusion were 16 cases and two levels were 3 cases. The mean operation time was 40 minutes per level with average bleeding 60 ml. Patients were asked to wear a hard collar for average 8.5 weeks after the operation. The fusion was solid on lateral view X-ray films three months post-operation. Only 1 case had neck stiffness due to prolonged collar protection and another case had short term pain on bone dorner site. There were no other complications such as cage subsidence, displacement, milaligment and kyphosis of cervical spine.

CONCLUSIONThe SynCage-C can resort and maintain the intervertebral space effectively with the advantage of immediate stability, satisfactory fusion and easy to use.

Adult ; Aged ; Cervical Vertebrae ; surgery ; Equipment and Supplies ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Orthopedic Procedures ; instrumentation ; methods ; Spinal Diseases ; surgery ; Spinal Fusion ; methods ; Titanium ; therapeutic use ; Treatment Outcome

Animals ; GAP-43 Protein ; metabolism ; Ganglia, Spinal ; drug effects ; metabolism ; Nerve Block ; Rats ; Rats, Wistar ; Sciatic Nerve ; drug effects ; injuries ; Tetrodotoxin ; pharmacology

Fibronectin containing extra domain A (EDA+ FN),a functional glycoprotein participating in several cellular processes,correlates with chronic liver disease.Herein,we aim to investigate the expression and secretion of EDA+ FN from hepatocytes in nonalcoholic fatty liver disease (NAFLD) and the underlying mechanisms.Circulating levels of EDA+ FN were determined by ELISA in clinical samples.Western blotting and flow cytometry were performed on L02 and HepG2 cell lines to analyze whether the levels of EDA+ FN were associated with endoplasmic reticulum (ER) stress-related cell death.Circulating levels of EDA+ FN in NAFLD patients were significantly higher than those in control subjects,and positively related with severity of ultrasonographic steatosis score.In cultured hepatocytes,palmitate up-regulated the expression of EDA+ FN in a dose-dependent manner.Conversely,when the cells were pretreated with 4-phenylbutyrate,a specific inhibitor of ER stress,up-regulation of EDA+ FN could be abrogated.Moreover,silencing CHOP by shRNA enhanced the release of EDA+ FN from hepatocytes following palmitate treatment,which was involved in ER stress-related cell damage.These findings suggest that the up-regulated level of EDA+ FN is associated with liver damage in NAFLD,and ER stress-mediated cell damage contributes to the release of EDA+ FN from hepatocytes.

OBJECTIVE: To explore the change of content of minor elements and its biological implication in apoptotic processes. METHODS: The content of minor elements of apoptotic cells from breast cancer induced by anti-cancer drugs was quantitatively analysed with synchronous radiation X-ray fluorescence. RESULTS: Seven kinds of minor elements including Ti, Cr, Fe, Cu, Zn, Ga and Ge as well as five major elements P, S, Ca, Cl and K were detected in the apoptotic and control cells. The content of elements Zn and P in apoptotic cells after 48 h treatment with taxol and colchicine was significantly increased higher than that in the control cells P<0.05). The content of element Fe in 48 h treatment with taxol was significantly decreased lower than that in the control cells P<0.05). The same trend of change of elements was observed during the process of apoptotic cells death induced with VP-16 or cycloheximide. CONCLUSION: This results suggest that the elements Zn, Fe and P should be involved in apoptosis induced by anti-cancer drugs.

OBJECTIVETo identify the relative factors of recent discovered atrial fibrillation (AF) following isolated coronary artery bypass grafting (CABG).

METHODSClassified the 649 cases undergoing isolated CABG from January 2005 to December 2006 to two groups according to whether AF appeared after operation. Collected the peri-operative data and operative strategy, then analyzed with single-factor analysis and Logistic regression.

RESULTSThe incidence of AF was 8.0% (52 cases), and 84.6% (44 cases) recovered sinus-rhythm leaving hospital. Age, standard European system for cardiac operative risk evaluation (EuroSCORE), ratio of high-operative-risk, left atrium diameter and ratio of left coronary artery dominance were higher in AF group than in non-AF group. Age, eject fraction, left atrium diameter, operative risk evaluation, left coronary artery dominance and anastomosis on right coronary artery were the relative factors of recent discovered AF following isolated CABG. But off-pump operation, prescription of adrenergic beta-antagonists pre-operatively and degree of coronary artery stenosis had no influence to AF.

CONCLUSIONSAF following CABG is a result of common influence by many factors. EuroSCORE might forecast partially the incidence of AF following CABG. Improve the myocardial protection and reduce the surgical damage during operative progress maybe the mostly approach to decrease the incidence of AF following CABG.

Aged ; Atrial Fibrillation ; etiology ; Coronary Artery Bypass ; adverse effects ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Postoperative Complications ; Risk Factors

OBJECTIVETo investigate the relationship between the protein expression of T-lymphoma invasion and metastasis gene 1 (Tiam-1) and the biological behaviors of nasopharyngeal carcinoma (NPC).

METHODSImmunohistochemistry was performed to detect the expressions of Tiam-1 protein in 60 specimens of NPC tissue, 20 specimens of chronic nasopharyngitis (CN) tissue, and 6 tumor tissues from nude mice inoculated with metastatic human NPC cells.

RESULTSThe positivity rate and average score for Tiam-1 expression were significantly higher in NPC tissues than in CN tissue (63.33% vs 36.67%, 2.9167 +/- 1.3057 vs 0.7000 +/- 0.9234; chi(2)=20.429, P=0.001; t=7.0162, P=0.0000, respectively). No difference was found in Tiam-1 expression among NPC patients in different T stages (F=2.36, P=0.0811), while the expression differed significantly between the patients with lymph node metastasis and those without metastasis, and also between patients with organ metastasis and those without (P=0.0001). High Tiam-1 expressions were found in the tumor tissues in nude mice inoculated with metastatic NPC cells.

CONCLUSIONTiam-1 expression is closely associated with the invasiveness and metastasis of NPC, indicating that Tiam-1 is an important factor that promotes the invasion and metastasis of NPC.

Adult ; Aged ; Animals ; Female ; Guanine Nucleotide Exchange Factors ; genetics ; metabolism ; Humans ; Male ; Mice ; Mice, Nude ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Transplantation ; T-Lymphoma Invasion and Metastasis-inducing Protein 1

AIMTo explore the changes of MMP-2/9 protein expression and excitation in brain of repetitive hypoxic mice.

METHODSThe biochemistry techniques of SDS-PAGE, Western bolt and Gel Goc Image Analysis System were applied to determine the level of MMP-2 and MMP-9 expression and activation in cortex and hippocampus of mice. The animals were randomly divided into 5 groups: the normal control group (H0), acute hypoxic (H1, hypoxic exposure once), repetitive hypoxic groups (H2-H4, repetitive hypoxia for 2-4 runs respectively).

RESULTS(1) The MMP- 2 expression level was increased first then decreased in hippocampus and the significant decrease was found in H4 group (P < 0.05, n=6), but no significant changes among the 5 groups in cortex. In addition, no activated form of 66 kD MMP-2 had been detected both in hippocampus and cortex. (2) Along with the development of brain hypoxic preconditioning, the level MMP-9 protein expression also increased first then decreased gradually in hippocampus, and the significant changes were found both in H1 and H4 groups (P < 0.05, n=7 for each group). The same trace of changes was also found in the activation of MMP-9 (include 82 and 78 kD forms) in hippocampus, and the significance both in H1 and H4 (P < 0.05, n=7 for each group) were detected. However, there was not any significant change in the level of MMP-9 protein expression or activation to be found in cortex.

CONCLUSIONThese results suggested that MMP-2 and MMP-9 might play certain role in the development of cerebral hypoxic preconditioning, the different changes of MMP-2/9 protein expression and activation both in cortex and hippocampus might be involved in their selective vulnerability to hypoxia.

Animals ; Hypoxia, Brain ; metabolism ; Ischemic Preconditioning ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Inbred BALB C