OBJECTIVE: To observe whether HSP70 could protect against H2O2-induced apoptosis in C2C12 myogenic cells by inhibiting Smac release from the mitochondria. METHODS: HSP70 gene and full length Smac gene was transiently transfected in C2C12 myogenic cells by lipofectamine and the protein levels of HSP70 and Smac were analysed by Western blotting. Hoechst 33 258 staining was used to examine cell morphological changes and to calculate percentage of apoptotic nuclei. DNA ladder pattern on agarose gel electrophoresis was used to observe the DNA fragmentation. Activities of caspase-3 and caspase-9 were assayed with Western blotting. The release of Smac from the mitochondria to the cytoplasm was observed by immunofluorescence. RESULTS: H2O2 ( 0.5 mmol/L ) activated caspase-3, caspase-9 8 h after the treatment and specific morphological changes of apoptosis 12 h after the treatment, and overexpression of Smac significantly promoted H2O2-induced activation of caspase-3, caspase-9 and apoptosis in C2C12 myogenic cells. HSP70 overexpression significantly inhibited H2O2-induced and Smac-promoted apoptosis, as shown by no specific DNA ladder pattern in agarose gel electrophoresis, decrease of percentage of apoptotic nuclei, and marked inactivation of caspase-3 and caspase-9. HSP70 could inhibit the release of Smac from the mitochondria to the cytoplasm 2 h after the treatment by H2O2. CONCLUSION HSP70 inhibits Smac release from the mitochondria and protects against H2O2-induced apoptosis in C2C12 myogenic cells.
Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; Cells, Cultured ; HSP70 Heat-Shock Proteins ; pharmacology ; Humans ; Hydrogen Peroxide ; Intracellular Signaling Peptides and Proteins ; metabolism ; Mitochondria, Heart ; drug effects ; metabolism ; Mitochondrial Proteins ; antagonists & inhibitors ; metabolism ; Myoblasts ; metabolism ; Myocytes, Cardiac ; drug effects ; metabolism

Objective - （ ） To evaluate the effect of job rotation on pain in wrist work related musculoskeletal disorders WMSDs （ ）Methods of physical therapists PTs . A total of 100 PTs from nine medical institutions were selected as the research subjects ， using judgment sampling method and they were divided into control group and intervention group by stratified random sampling ， method with 50 person in each group. The individuals in control group perform routine works. People in the intervention group were rotated between posts or added mobile shift replacements in daily work for 30 minutes. The duration of intervention was ， ， （ ） once a day five days a week for ten weeks. Visual Analogue Scale VAS score and pain duration were used as the evaluation ， indexes of intervention effect. The changes of indexes before intervention five weeks and ten weeks after intervention were Results ， compared between the two groups. Before intervention there was no significant difference in the VAS score and pain （ P ） duration between the control group and the intervention group all >0.05 . There was no significant difference in VAS score （ P ） and pain duration among the control group at three time points after intervention all >0.05 . The VAS score of PTs in the （P ）， intervention group at ten weeks was lower than that in the control group at the same time point <0.05 and it was lower than （ P ） that before intervention and at five weeks of intervention in the same group all <0.05 . The pain duration of PTs in the （ P ）， intervention group was lower than that in the control group at five and ten weeks after intervention all <0.05 and was lower （ P ） Conclusion ， than that before intervention at the same group all <0.05 . Rotating schedule can relieve WMSDs of PTs and the effect of intervention for ten weeks is more effective than that of intervention for five weeks.

Objective To study the relationship between environmental chemical elements,vires infection and dilated cardiomyopathy(DCM).Methods In 2008,233 patients with DCM(case group)and 150 patient with stable angina(control group)were chosen in Liaocheng People's Hospital and Yanggu People's Hospital,Shandong province.Population distribution and disease history were surveyed in the two groups.Human myocardial antibody IgG(AMA-IgG),Coxsackie B virus IgG(CBV-IgG),Adenofirus antibody IgG(ADV-IgG)were detected by ELISA in both the case group and the control group.Serum trace elements were detected in the two groups.The general chemical and toxicological indicators in drinking water of the high-and the low-incidence aireas of the disease were control group[60.00%(90/150),χ2=13.80,P<0.01)].Per capita annual income(Yuan,RMB)in the case group (3207.82±618.51)was lower than that of the control group[(5086.61±886.12),t=24.40,P<0.01].Personal alcohol consumption in the case group[(175.00±160.50)g/d]was higher than that of the control group[(110.22±100.03)g/d,t=4.40,P<0.01)].The rate of myocarditis in the case group[5.15%(12/233)]was higher than ADV-IgG in the cage group were 7.78%(7/90),6.67%(6/90)and 6.67%(6/90),respectively.Compared with those in the control group[3.33%(2/60),5.00%(3/60)and 5.00%(3/60),χ2=1.26,0.18,0.18,all P>0.05],no mg/L]in drinking water of the high-incidence areas were significantly higher than that of iron[(0.39±0.67)mg/L,t=2.11,P<0.05]and that of manganese[(0.15±0.14)mg/L,t=3.01,P<0.01]in the low-incidence arefas.The content of semm iron[(69.1±57.8)μmol/L]in the case group evidently exceeded the normal range(15.6-35.9 μmol/L)and obviously higher than that in the control group[(20.0±17.5)μmol/L,t=5.04,P<0.01].Conclusions Theso data do not support that DCM is related with persistent virus infection and autoimmunization.DCM is probably related with low incomes,high alcohol consumption,myocarditis,high iron and manganese contents in drinking water and high content of serum iron.

OBJECTIVE: To evaluate influences of regular-dose of adriamycin (ADR) on heart function and sarcoplasmic reticulum (SR) Ca2+ -ATPase in cardiomyocyte of rabbits. METHODS: Nine rabbits received intraveneous injection of ADR (2mg/kg) once a week for 8 weeks, the rabbits injected with saline were used as control group. Cardiac output (CO), blood pressure (BP), mean artery pressure (MAP), left ventricular systolic pressure( LVSP), left ventricular end-diastolic pressure (LVEDP), calcium in cardiomyocyte (MyoCa2+) of rabbits and SR Ca2+ -ATPase activity were examinated 3 weeks after the final injection. RESULTS: CO, LVSP and SR Ca2+ -ATPase activity were significantly decreased in ADR treated group compared with the control group. Conversely, LVEDP and MyoCa2+ were significantly increased in ADR treated rabbits. CONCLUSION: Heart function can be decreased by regular-dose of ADR in injection. Calcium overload in cardiomyocyte and decrease of SR Ca2+ -ATPase activity is important physiopathologic mechanism in ADR-induced impairment of heart.

BACKGROUNDThe fat derived protein adiponectin plays an important role in the regulation of glucose metabolism. The aim of this study was to provide the experimental basis for further investigating on adiponectin (ADPN) function. Its eukaryotic recombinant was constructed and expressed in precursor cells of 3T3-L1 adipocytes. The effects of dexamethasone on peroxisome proliferator activated receptor-gamma (PPAR-gamma) mRNA expression in 3T3-L1 cells with human recombinant adiponectin were assessed.

METHODSThe recombinant plasmid pMD18-T-hADPN and eukaryotic expression vector pcDNA3.1(+) were digested by two restrictive endonucleases and adiponectin and linear pcDNA3.1(+) were obtained. Then, they were ligated and translated into JM109. The recombinant pcDNA3.1(+)-hADPN so obtained was identified by digestion by restrictive endonuclease and nucleotide sequencing. The 3T3-L1 precursor cells were transfected using SuperFect Transfection Reagent (Qiagen). Furthermore, 3T3-L1 cells with human recombinant adiponectin incubated with dexamethasone (0.5 mmol/L) for 24 hours, cells were collected and total RNA was extracted. The PPAR-gamma mRNA expression was quantified by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSAfter eukaryotic recombinant was digested by Hind III and EcoR I, fragments of 800 bp and 5.4 kb were identified by nucleotide sequence scanning and consistent with theoretical values. Electrophoretogram of RT-PCR in 3T3-L1 precursors showed only one band in front of 250 bp, which was consistent with theoretical value 234 bp. In the 3T3-L1 cells, 3T3-L1 cells with plasmid and 3T3-L1 cells human recombinant adiponectin, treatment with dexamethasone (0.5 mmol/L) decreased PPAR-gamma mRNA expression compared to untreated controls (P < 0.01). Effect of dexamethasone on PPAR-gamma mRNA expression in 3T3-L1 cells was reversed by stably transfected human recombinant adiponectin.

CONCLUSIONThe 3T3-L1 cells stably transfected human recombinant adiponectin had increased PPAR-gamma mRNA expression. Dexamethasone suppressed PPAR-gamma mRNA expression in the 3T3-L1 cells. Effect of dexamethasone on PPAR-gamma mRNA expression in 3T3-L1 cells was reversed by stably transfected human recombinant adiponectin.

3T3-L1 Cells ; Adiponectin ; physiology ; Animals ; Dexamethasone ; pharmacology ; Insulin Resistance ; Mice ; PPAR gamma ; genetics ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVEAccording to the information obtained from the revision of total hip arthroplasty, the situations of the artificial femoral stem under physiological stress were analyzed preliminarily. In order to accumulate clinical experience, the discussions about how to enlongate the life of artifical joints were performed.

METHODSFifty-three patients required revision operations were selected, including 28 males and 25 females,with an average age of 74.66 years old (61 to 84 years old). The average life of artificial joints was 18.24 years (3 to 27 years). The initial weightbearing was started 11 days (5 to 16 days) after the first operation. The main reasons for the revision were the spontaneous loosening and trauma. Based on imaging and surgical findings, as well as the histological pathology, the situations of the stems in human bodies were determined.

RESULTSFemoral prosthesis would sink under physiological stress. The design of femoral stems, motion of patients', and bone growth determined the secondary stability. Straight stems were helpful for physiological subsidence of prosthesis, achieving the stability with close bone-implant contact by bone growth,which allowed the patient to do early landing exercise. The collared prosthesis and prosthesis combined with internal fixation limited the subsidence of femoral stems. Their stability depends on the time of initial placement and the bone growth for supporting. Delaying the inital weightbearing was suggested for patients with these protheses.

CONCLUSIONAccording to the actual situation of the patients, the appropriate chosen of femoral stems and time to perform the weightbearing can extend the life of the femoral prosthesis.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; adverse effects ; methods ; Female ; Hip Prosthesis ; Humans ; Male ; Middle Aged ; Prosthesis Design

OBJECTIVETo study the bone state with ultimate stress by examining biomechanical distribution of upper femur in Chinese, in order to accumulate more experiences for clinical work.

METHODSTotally 60 Chinese femurs from fresh cadavers were randomly divided into two groups. All the femurs were cleaned, and the body age ranged from 36 to 72 years old, averaged 56.4 years, including 41 from males, and 19 from females. These two groups underwent mechanical stress and bending stress tests. Special mechanical laboratory and machines were used to get the information. Results about the loading value at each testing point under stress were collected.

RESULTSThe four faces of the upper femur suffered different stress under external forces. The bone on upper femur can tolerate more mechanical stress than bending stress. Medial and lateral region of the femur neck and the rear side of the small tuberosity section were themain position enduring the vertical stress. The rear position of the base femur neck and the small tuberosity section were the main regions enduring the bending stress. Those main positions had strong cancellous bones. The intertrochanteric fracture fixation and artificial femoral stems were designed depending on this biomechanical basis.

CONCLUSIONAccording to our experiment result, doctors need to chose more effective fixations for upper femur fracture, and femoral stems for the patients. More information should be collected by further researches.

Adult ; Aged ; Biomechanical Phenomena ; Female ; Femur ; chemistry ; Humans ; Male ; Middle Aged ; Stress, Mechanical

Drug delivery system (DDS) is a novel approach to overcome multidrug resistance (MDR) in tumors nowadays. This work was designed to investigate a new micellar delivery system for in vitro reversal of resistant ovarian tumor cells, based on a nonionic triblock copolymer Pluronic P105 and paclitaxel (PTX). The PTX-loaded polymeric micelles (P105/PTX) were prepared by thin film-hydration methods. Based on the results of single factor experiments, the P105/PTX micelle formulation was optimized by employing the central composite design-response surface methodology. The physico-chemical properties of the P105/PTX micelles were characterized, including micelle size, drug loading coefficient, in vitro release behavior, etc. The cytotoxicity of the P105/PTX micelles was assessed against human ovarian tumor cell line, SKOV-3/PTX, by a standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl (MTT) assay. In order to understand the possible mechanism of Pluronic effects in resistant tumor cells, cellular uptake study of micellar PTX or Rhodamine-123 (R-123) was also carried out. The results showed that the micelle size was about 24 nm with drug loading coefficient of 1.1% and PTX concentration of 700 microg x mL(-1). The cumulative release amount of PTX from the P105/PTX micelles was only 45.4% in 6 h (P < 0.05) and 79.6% in 24 h, whereas Taxol injection in 6 h released 95.2% PTX. The IC50 values of the P105/PTX micelles and Taxol injection against SKOV-3/PTX were 1.14 and 5.11 microg x mL(-1), and resistance reversion index (RRI) was 9.65 and 2.15, respectively. The micellar PTX or R-123 exhibited a significant increase in cellular uptake in resistant SKOV-3/PTX cells compared with free PTX or R-123. These results indicated that PTX could effectively be solubilized by Pluronic P105 block copolymers via thin film-hydration process and formulation optimization, producing nano-scale polymeric micelles with sustained release property in vitro. The P105/PTX micelles were effectively able to reverse resistance to PTX in SKOV-3/PTX tumor cells compared with Taxol injection or free PTX solution, and the enhanced cytotoxicity in the resistant SKOV-3/PTX cell was related to the improved cellular uptake of PTX by Pluronic P105 copolymers.
Antineoplastic Agents, Phytogenic ; administration & dosage ; chemistry ; pharmacology ; Cell Line, Tumor ; Drug Carriers ; Drug Delivery Systems ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Excipients ; chemistry ; Female ; Humans ; Inhibitory Concentration 50 ; Micelles ; Ovarian Neoplasms ; metabolism ; pathology ; Paclitaxel ; administration & dosage ; chemistry ; metabolism ; pharmacology ; Particle Size ; Poloxamer ; chemistry

OBJECTIVE: To determine the changes of advanced oxidation protein products (AOPP) in diabetic nephropathy (DN) patients, as well as its relationship with superoxide dismutase (SOD), glutathione peroxidase (GPx) and neopterin (NPT). METHODS: By the concentration of urinary albumin excretion rate (UAER) and creatinine (Cr), 85 diabetes mellitus (DM) patients were divided into 4 groups as non-DN group (DM), early-staged DN group (DN3), significant DN group (DN4) and end-staged DN group (DN5). The concentration of the serum AOPP was measured by ameliorated method introduced by Wikto-Sarsat, while SOD by Xanthine oxidase test, GPx by [5,5'Dithio-bis (2-Nitrobenzoic aicd) ] (DTNB) reaction test and NPT by ELISA. RESULTS: AOPP in Group DN5 [(117.8 +/- 64.8) [micromol/L] and Group DN4 [ (80.0 +/- 23.0) micromol/L] were significantly higher than those in Group DM [(58.2 +/- 17.7) micromol/L]. There was no significant difference of AOPP between Group DN3 [(72.7 +/-17.2) micromol/L] and Group DM. Serum AOPP negatively correlated with SOD and GPx (r = -0.217 and -0.374 respectively, P < 0.05), while positively correlated with NPT (r = 0.499, P < 0.01). CONCLUSION DN patient has enhanced protein oxidation than DM patient, which is related to oxidative stress and chronic inflammation in DN.
Aged ; Diabetes Mellitus, Type 2 ; blood ; Diabetic Nephropathies ; blood ; Female ; Glutathione Peroxidase ; blood ; Humans ; Male ; Middle Aged ; Neopterin ; blood ; Oxidation-Reduction ; Oxidative Stress ; Superoxide Dismutase ; blood

OBJECTIVE: To observe the effect of Krüppel-like factor 4 (KLF4) overexpression on heat stress-induced apoptosis of Raw264.7 macrophages. METHODS: The fragment containing full length mouse KLF4 cDNA coding sequence was inserted into the pcDNA3.1 vector and Raw264.7 macrophages were transfected with pcDNA3.1-KLF4 plasmids using lipofectamine.The expression of KLF4 was examined by Western blot in the Raw264.7 macrophages stably transfected with pcDNA3.1- KLF4 plasmids. Raw264.7 cells transfected with pcDNA3.1 and pcDNA3.1-KLF4 were exposed to heat stress (42 degrees C) for 1h and recovered at 37 degrees C for 12h. Flow cytometry, Hoechest 33258 staining assay, and DNA ladder assays were performed to assess the apoptosis. RESULTS: The KLF4 overexpressed Raw264.7 macrophages were established. After the heat stress, flow cytometry showed that apoptotic cells increased significantly in KLF4 overexpressed cells compared with the vector control; Hoechest 33258 staining was characterized with classical changes including apoptotic body, and nuclear condensation. Agarose gel electrophoresis showed that "DNA ladder" could be observed clearly. CONCLUSION KLF4 overexpression can increase heat stress-induced apoptosis of Raw264.7 macrophages.
Animals ; Apoptosis ; genetics ; Cell Line ; Genetic Vectors ; Heat-Shock Response ; Kruppel-Like Transcription Factors ; genetics ; Macrophages ; cytology ; Mice ; Plasmids ; Transfection