Objective To observe the effect of autophagy on paclitaxel-induced CaSki cell death through the regulation of the expression of autophagy gene Beclin1, and to explore the interaction and relationship between autophagy and apoptosis. Methods Eukaryotic expression vector pcDNA3.1-Beclin1 and RNA interference vector pSUPER-Beclin1 were transfected into human cervical cancer CaSki cells in vitro and screened for stable expression cell lines. The formation of autophagic vacuoles was observed with an electronic microscope. The expression of Beclin1 and LC3 was measured by Western blot. After being treated with paclitaxel, the change of cell proliferation was assessed by MTT assay, the percentage of apoptotic cells and autophagic cells were analyzed by flow cytometry. Results A lot of autophagic vacuoles were observed in pcDNA3.1-Beclin1 cells by electronic microscopy. Beclin1 and LC3 protein expression was up-regulated in CaSki cells transfected with pcDNA3.1-Beclin1, and was inhibited in cells transfected with pSUPER-Beclin1. MTT assay revealed the survival rate of CaSki cells was significantly decreased after being transfected with pcDNA3.1-Beclin1. After being treated with paclitaxel, the percentages of apoptotic cells and autophagic cells were both increased in pcDNA3.1-Beclin1 group compared with that of the blank control group especially the increase of apoptosis was particularly evident. Conclusion Autophagy and apoptosis have different roles in the process of paclitaxel-induced cervical cancer CaSki cell line death. Overexpression of Beclin1 in CaSki cells may enhance the apoptosis induced by paclitaxel.

Objective To investigate the effect of different Chinese herbs on cell proliferation and cartilage oligomeric matrix protein(COMP)expression in chondrocyte culture.Methods Chondrocytes isolated from rabbit knee cartilage were cultured for 3 generations with the density of 2?10~4/cm~2 and were verified by collagenⅡimmunohistochemical staining.Rabbit sera containing herbs were obtained after animals orally ad- ministrated herbs at the dosage equivalent to human.At 5% and 10% serum density,cells were cultured in the medium that contained liver-softening herbal compound sera.Subgroups setting at 1,3 and 5 hours after herb intervention were observed.Rabbit and bovine sera were control groups.Seven days after intervention,chon- drocytes proliferation was observed using the MTT assay kit.For the study of COMP expression,chondrocytes were isolated from human knee cartilage supematant.Superuatant COMP level was tested by enzyme-linked immunoabsorbent assays(ELISA)after directly adding compound and extract from liver-softening herbs to the culture at the final concentration of 10 mg/ml for 3 days.Results Liver-softening herbal compound group had significant effect on cell proliferation compared to control,of which,3-hour subgroup was more significant than 1-and 5-hour subgroups(P

OBJECTIVETo explore the inhibition and molecular mechanism of icaritin (ICT) combined doxorubicin (DOX) on human osteosarcoma MG-63 cells in vitro.

METHODSThe control group, ICT groups (10, 20, 40, 80, and 160 µmol/L), DOX groups (1, 2, 4, 8, and 16 µg/mL), and combination groups (20 µmol/ L ICT +1 µg/mL DOX, 20 µmol/L ICT +2 µg/mL DOX, 20 µmol/L ICT +4 µg/mL DOX, 40 µmol/L ICT +1 µg/mL DOX, 40 µmol/L ICT +2 µg/mL DOX, 40 µmol/L ICT +4 µg/mL DOX, 80 µmol/L ICT +1 µg/mL DOX, 80 µmol/L ICT +2 µg/mL DOX, 80 µmol/L ICT +4 µg/mL DOX) were set up. Human osteosarcoma MG-63 cells were respectively cultured and their effects on morphological changes were observed using inverted phase contrast microscope after 24-and 48-h intervention. The cell proliferation inhibition rate of each group was de- termined using CCK-8, and IC50 calculated. The MG-63 apoptosis rate was detected using Annexin V-FITC/ PI double dye flow cytometry. Expression levels of bcl-2, caspase-3, and p21 were detected using RT-PCR.

RESULTSICT and DOX could obviously inhibit the proliferation of MG-63 cell. Along with ICT concentration increasing from 10 µmol/L to 160 µmol/L, the cell proliferation inhibition rate also increased gradually from 9.67% ± 3.62% to 89.18% ± 9.66%. The IC50 was 46.93 µmol/L and 3.87 µg/mL respectively. ICT and DOX could cause either early or late stage apoptosis, down-regulate Bcl-2 gene expression, and up-regulate gene expressions of Caspase-3 and p21 respectively (P < 0.05). Aforesaid changes were more obviously seen in combination groups than in lCT groups and DOX groups (P < 0.05).

CONCLUSIONCT combined DOX had additive or synergistic inhibition effect for the proliferation of osteosarcoma MG-63 cells, which might be related with regulating gene expressions of bcl-2, caspase-3, and p21.

Apoptosis ; Bone Neoplasms ; metabolism ; pathology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Down-Regulation ; Doxorubicin ; pharmacology ; Drug Synergism ; Flavonoids ; pharmacology ; Humans ; Osteosarcoma ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism

Objective:To investigate frequency of blood stasis syndrome(BSS) defined by traditional Chinese medicine in cerebral infarction and its correlations with carotid atherosclerotic plaque(CAP).Methods: All subjects comprised 151 patients aged 40 to 80 years(Mean ? SD age,65 ?11 years) with 67.9% for males and 32.1% for females.With the use of ACUSON7 color Doppler ultrasound,carotid atherosclerosis was evaluated by the plaque score,the left plaque score,the right plaque score,the numbers of the plaque respectively as defined by the sum of all plaque heights in bilateral carotid arteries.On the basis of neurological signs and symptoms,medical history,and brain MRI,we diagnosed stroke and its subtypes as follows: stroke(n=117),and vertebrobasilar insufficiency(VBI)(n=34) without the history of the stroke,which were based on Diagnostic Criteria for Cerebral Vascular Diseases in 2005.Diagnosis for syndromes defined by traditional Chinese medicine were made according to Diagnostic Criteria for Stroke in 1994.One-way ANOVA was used in comparison between groups,and multivariant Logistic Regression Analysis was conferred in correlations between several variables.Results: 47.0% of all cases with cerebral infarction presented the BSS,with as lower than syndrome of fire-heat(51.0%),as but significantly higher than syndrome of Qi deficiency(32.0%),liver-wind syndrome(27.0%),phlegm syndrome(23.0%) and syndrome of asthenic yin causing predominant yang(6.0%).There is a significant difference between groups for 44(79.0%) cases of 56 patients with cerebral infarction and the BSS have CAP,and only 35(57.0%) cases of 61 patients with cerebral infarction but without the BSS have CAP(P

Objective:To establish an LC-MS/MS method for the content determination of triptolide in Shenshe ointment.Methods:The chromatographic separation was performed on an Agilent TC-C18 (250 mm×4.6 mm,5 μm)column with the mobile phase consisting of methanol-0.1% formic acid solution (73∶27,v/v).The flow rate was 0.5 ml·min-1 and the column temperature was maintained at 35℃.An electrospray ionization (ESI) source was applied with multiple reaction monitoring (MRM) combined with a positive mode.Metronidazole was used as the internal standard.The mass transitions were 361.4→43.2 for triptolide and 172.1→128.0 for metronidazole,respectively.Results:The linear range was 10-500 ng·ml-1 with good correlation coefficient (r=0.999 9).The limit of quantification (LOQ) was 9.5 ng·ml-1.The intra-and inter-day RSDs of peak areas were all less than 3% and the average recovery was 96.87%(RSD=2.79,n=6).Conclusion:The established LC-MS/MS method is simple,efficient,sensitive and accurate in the quality control of Shenshe ointment.

Objective:To summarize our experience in perioperative anesthetic management for fulminant hepatic failure (FHF)patients receiving liver transplantation.Methods:The clinical anesthetic data of 48 FHF patients receiving orthotopic liver transplantations(OLT)from January 2006 to January 2007 were retrospectively analyzed,and the anesthetic management expe- rience was summarized.General anesthesia was applied;the hemodynamics was monitored during the operation and doses of adrenaline and phenylephrine were adjusted according to the monitoring results.Blood samples were obtained before operation, before anheptic,30 min after anhepatic phase,5 min before neohepatic phase,and 5 min,30 min and 60 min after neohepatic phase for blood gas and electrolyte analysis and for determination of coagulation function;the drugs were subsequently adjusted according to analysis results.Results:All the 48 patient underwent successful anesthetic management and there was no death dur- ing opearation.The average blood loss during operation was(5 219?478)ml.Mild alkalosis,hypokalemia,hyponatrium,and hy- pocalcemia were present before operations,pH,BE and HCO_3~- were obviously reduced 30 min after anhepatic phase and in- creased 60 min after neohepatic phase.Kalemia was obviously increased 30 min following anhepatic phase and began to increase 60 min following neohepatic phase.Calium concentration was decreased at the end of preanhepatic phase(P

Animals ; Cell Line, Tumor ; Cell Survival ; drug effects ; Ferrosoferric Oxide ; adverse effects ; metabolism ; pharmacology ; Humans ; Liver ; chemistry ; drug effects ; Lung ; drug effects ; metabolism ; Magnetics ; Mice ; Nanoparticles ; adverse effects ; Spectrophotometry, Atomic

OBJECTIVETo understand the information of female patients with knee osteoarthritis regarding muscle force, constitution parameter.

METHODSThirty-seven cases diagnosed as knee osteoarthritis and 37 controls were examined by MES. T-test was used to analysis two groups differences of muscle force, constitution parameter, et al.

RESULTSCompared between affected limbs and controls limbs in patients revealed that the lower limb muscle distribution index of the affected limbs was higher than the control limbs (P<0.05), but comparison in functional status the lower limb muscle force, muscle functional index and muscle force of unit volume of the affected limbs were lower than the control limbs (P<0.05). Compared between patients group and control group the muscle force of both lower limbs, muscle functional index and muscle force of unit volume were lower than control group (P<0.001).

CONCLUSIONThe utility muscle force of lower limbs of female patients with knee osteoarthritis is weaker than healthy female. Muscle function disorder instead of muscle atrophy is the key cause of the weakness.

Adult ; Aged ; Body Composition ; Case-Control Studies ; Female ; Humans ; Middle Aged ; Muscle Contraction ; Muscle, Skeletal ; metabolism ; physiopathology ; Osteoarthritis, Knee ; metabolism ; physiopathology ; Range of Motion, Articular

OBJECTIVETo observe the distribution features of tender points in knee of patients with knee osteoarthritis in order to provide evidences for the treatment and diagnosis.

METHODSFrom November 2011 to December 2012,86 patients with knee osteoarthritis were recruited, including 21 males and 65 females, ranging in age from 45 to 85 years old, with an average of (59.98 +/- 8.23) years old. The course of disease ranged from 3 months to 15 years. The tender points and its distributions were determined by finger press carefully on their knees. Data of studying was analyzed by frequency statistics and Hierachical cluster analysis.

RESULTSThe distribution of tender points in the knee osteoarthritis was mainly in the interior region and anterior area such as in apex of patella, adductor tubercle and et al. According to the results of hierachical cluster analysis, the tender points could be divided into two categories the first cluster was in the interior region of knee, the second cluster was in the lateral region.

CONCLUSIONThe findings demonstrated that cluster analysis statistical method can be used for classification of the distribution of tender points. The distribution features of tender points in knee osteoarthritis are related to the anatomic site in knee.

Adult ; Aged ; Aged, 80 and over ; Cluster Analysis ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; complications ; Pain ; complications

Osteoarthritis (OA) is the most prevalent of joint diseases,and its pathology is characterized by the degeneration of cartilage, sclerosis of subchondral bone, and osteophyte formation. Localization of the early lesions of OA has not been clarified, but many researchers have focused on cartilage and have considered that changes in subchondral bone occur subsequently to the degeneration of cartilage. However, a low bone mineral density, particularly in the knee joint with OA, high bone turnover, and efficacy of bone resorption inhibitors for OA have recently been reported, suggesting that subchondral bone plays an important role in the pathogenesis of OA. This review aims to make a conclusion about advancement in research of subchondral bone in osteoarthritis.
Bone Remodeling ; Bone and Bones ; pathology ; Cartilage ; pathology ; Humans ; Osteoarthritis ; etiology ; pathology ; physiopathology