Mechanical loading is the main "instructive" factor of bone formation. The mechanism becomes the heat point in the field of bone science, biomedicine project and rehabilitation research. Mechanical vibration is one of the mechanic stimulation. Evidences show that vibration has obvious anabolic effect, and will have a broad landscape in the treatment of osteoporosis. But in the present studies, there were incorporated with the vibration frequency, and intensity, in particularly, few reports about the mechanism of vibration to bone. It will provide theoretic foundation for further systematic, sound research and its clinical application.
Animals ; Biomechanical Phenomena ; Bone Diseases ; physiopathology ; therapy ; Bone and Bones ; physiology ; physiopathology ; Humans ; Musculoskeletal Manipulations ; Osteogenesis ; Vibration ; therapeutic use

Objective To study the effect of dynamic stress stimulation on the expression of Sox9 mRNA and protein in metaphyseal chondrocytes in vitro, and to explore the specific mechanism of mechanical signal transduction. Methods The rat metaphyseal chondrocytes separated and cultured for the 3rd generation in vitro were randomly divided into four groups:control group (all interventions were not applied), simple dynamic pressure group (a dynamic pressure stimulus with a size of 90 mmHg and a frequency of 0.1 Hz was applied using an open pressure control culture system), simple calcium antagonist group (the concentration of 10μmol/L nifedipine was given) and dynamic pressure+calcium antagonist group (a dynamic pressure stimulus with a size of 90 mmHg, frequency of 0.1 Hz and concentration of 10 μmol/L nifedipine were given at the same time). The expression of Sox9 mRNA was detected after 24 h intervention by real-time quantitative polymerase chain reaction (RT-PCR) in four groups. The expression of Sox9 protein was detected by Western blot assay. The intracellular free Ca2+ in metaphyseal chondrocytes was labeled with Fluo-3/AM, and the average fluorescence intensity detected by laser scanning confocal scanning microscopy was compared between four groups. Results The expression of Sox9 mRNA was 3.81 times higher in dynamic stress group than that in the control group, and the protein expression level was 2.33 times higher than that of the control group (P<0.05). There were no significant differences in the expression of Sox9 mRNA and protein between the calcium antagonist group and the control group. The expressions of Sox9 mRNA and protein were lower in dynamic pressure+calcium antagonist group than those in the dynamic stress group, but which were higher than those of control group(P<0.05). The results of average fluorescence intensity showed that there was no significant difference in the intracellular free Ca2+ concentration between four groups (P > 0.05). Conclusion Dynamic stress stimulation can increase the expression of Sox9 mRNA and protein in rat metaphyseal chondrocytes. There is calcium channel involvement in the mechanical signal transduction.

OBJECTIVETo investigate synergistic killing effect of anti-human DR5 (death receptor 5 of TRAIL) monoclonal antibody (mDRA-6) and adriamycin(Adr) on HL-60 cells.

METHODSmDRA-6 was prepared by immunizing BALB/c mice with DR5 protein. DR5 expression on Adr-treated HL-60 cells was detected by flow cytometry. Morphologic changes of HL-60 cells were observed under fluorescence microscope. Cytotoxic and apoptotic effects of mDRA-6 and Adr on HL-60 cells were measured by MTT analysis. DNA fragmentation was detected by agarose gel electrophoresis.

RESULTSAdr induce DR5 expression on HL-60 cells. Cell budding, chromatin condensation and apoptotic body formation were observed in HL-60 cells treated by mDRA-6 and Adr. Death and apoptosis of these cells and DNA ladder were exhibited on agarose gel electrophoresis.

CONCLUSIONmDRA-6 and Adr have synergistic killing effect on HL-60 cells.

Animals ; Antibodies, Monoclonal ; pharmacology ; Apoptosis ; drug effects ; Dose-Response Relationship, Drug ; Doxorubicin ; pharmacology ; HL-60 Cells ; Humans ; Mice ; Mice, Inbred BALB C ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; immunology ; TNF-Related Apoptosis-Inducing Ligand ; immunology

Objective To study the effect of dynamic pressure on the expression of parathyroid hormone-related protein (PTHrP)mRNA in metaphyseal cartilage stem cells of rats so as to further explore whether fiber actin (F-actin)is involved in the mechanical signal transduction process.Methods We isolated and cultured metaphyseal cartilage stem cells of rats by immunomagnetic beads.The third-generation rat metaphyseal cartilage stem cells were randomly divided into four groups:0%,3%,6%,and 12% deformed groups according to the size of dynamic pressure strength.We used a self-prepared dynamic tonic culture device to exert different intensity of pressure on each group of cells for 24 hours.Flow cytometry was used to detect the cell cycle distribution and apoptosis rate.The expression of PTHrP mRNA in each group was detected by Rea-l time quantitative PCR. Furthermore,the third-generation rat metaphyseal cartilage stem cells were randomly divided into four groups:control group,simple pressure group (6% deformation),pressure+cytoskeleton relaxin D group,and simple cytoskeleton relaxin D group according to whether or not to apply pressure and cytoskeleton relaxin D.F-actin fibers in each group of cells were stained with phalloidin and placed under a laser scanning confocal microscope.The expression of PTHrP mRNA in each group was detected by Real-time quantitative PCR.Results The results of flow cytometry showed no significant difference in G0/G1,G2/M and S phases between 0%,3%,6% and 12% deformed groups (P＞0.05).There was no significant difference in the apoptosis rate between 3% and 6% deformed groups compared with 0% deformed group (P＞0.05).The apoptosis rate was significantly higher in 1 2 % deformed group than in control group (P＜0.05).The results of laser confocal microscopy showed that the arrangement of F-actin fibers in the pressure group was neat and parallel compared with that in the control group, which was consistent with the direction of force.The intracellular F-actin fiber structure in pressure+cytoskeleton relaxin D group and simple cytoskeleton relaxin D group was destroyed and aggregated into clusters.Real-time quantitative PCR results showed that PTHrP mRNA expression did not significantly differ between 3% and 0% deformed groups (P＞0.05).The expression of PTHrP mRNA in 6% and 12% deformed groups was significantly higher than that in 0% group (P＜0.05).The expression of PTHrP mRNA in the cells of simple pressure group was significantly higher than that in the control group (P＜0.05).There was no significant difference in the expression of PTHrP mRNA between simple cytoskeleton relaxin D group and control group (P＞0.05).The mRNA expression of PTHrP was higher in pressure+cytoskeleton relaxin D group than that in control group,but lower than in simple pressure group (P＜0.05).Conclusion The dynamic pressure of proper intensity can increase the mRNA expression of PTHrP in chondrocytes of metaphyseal hypertrophy in rats,and F-actin is involved in the mechanical signal transduction process.

Objective To analyze the related pathogenicity gene mutations in a sudden death of hypertrophic cardiomyopathy (HCM) on whole exome level.Methods Whole exome sequencing (WES) was been performed on a sudden death case sample with pathological features of HCM by Illumina(R) Hiseq 2500 platform.Using hgl9 as the reference sequences,the sequencing data were analyzed.Suspicious single nucleotide variants (SNV) were screened,and the conservatism and function were analyzed by the software such as PhyloP,PolyPhen-2,SIFT,etc.Results After screening,a heterozygous mutation C719R was finally identified in the gene MYBPC3 of this case.Conclusion The molecular anatomy on whole exome level by second generation sequencing technology can help to define the molecular mechanism of HCM and provide a new mothed and thought for analysis of death cause.

Refractory gelastic seizure is one of the most common clinical manifestations in patients with hypothalamic hamartoma (HH) and HH is usually regarded as the epileptogenic focus. A young female patient with a small HH and refractory seizures is reported here. However, both the seizure semiology and results of electroencephalogram monitoring indicated the right temporal region was the epileptogenic focus. Thus a standard right anterior temporal lobectomy was performed while the hamartoma preserved. There was a marked improvement in both seizure frequency and quality of life during a 13-month follow-up. The outcome supported the concept that independent epileptogenic focus outside of the hypothalamus might occur in patients with HH.
Adult ; Electroencephalography ; Epilepsy, Temporal Lobe ; diagnosis ; surgery ; Female ; Hamartoma ; diagnosis ; surgery ; Humans ; Hypothalamic Diseases ; diagnosis ; surgery

OBJECTIVETo investigate the effects of dopamine and norepinephrine on the renal function in the patients with septic shock.

METHODSEighty-seven patients with septic shock were divided into three groups (group A, B, C) according to the biggest infusing rate of norepinephrine, with the infusing rate of 0.5 - 0.9, 1.0 - 1.5, 1.6 - 2.0 microg x kg(-1) x min(-1), respectively. Mean arterial blood pressure (MAP), heart rate (HR), urine output, blood urea nitrogen (BUN), creatinine (CRE), urine albumin (U-ALB) and urine beta(2)-microglobulin (Ubeta(2)-MG) as well as APACHE III score in all the patients were detected.

RESULTSBefore anti-shock therapy was given, hypotension, tachycardia and oliguria occurred in all the 87 patients, and CRE, BUN, U-ALB, Ubeta(2)-MG and APACHE III score were abnormal in most cases. With the anti-shock therapy, MAP, HR, urine output and BUN, CRE in all patients returned to normal levels gradually, and U-ALB, Ubeta(2)-MG levels and APACHE III score also restored but still remained abnormal.

CONCLUSIONSThe first aim of treating septic shock should be restoring the organ blood supply, and based on volume resuscitation, dopamine, noradrenaline and other vasoactive drugs could be combined to maintain circulatory stability.

APACHE ; Adult ; Aged ; Blood Transfusion ; Cardiotonic Agents ; administration & dosage ; Combined Modality Therapy ; Dopamine ; administration & dosage ; Drug Therapy, Combination ; Female ; Humans ; Kidney ; drug effects ; physiopathology ; Male ; Middle Aged ; Norepinephrine ; administration & dosage ; Retrospective Studies ; Shock, Septic ; physiopathology ; therapy ; Vasoconstrictor Agents ; administration & dosage

Humans ; East Asian People ; External Fixators ; Fracture Fixation/methods* ; Fracture Fixation, Internal/methods* ; Fractures, Bone/surgery* ; China ; Minimally Invasive Surgical Procedures/methods*

Betacoronavirus ; physiology ; Coronavirus Infections ; drug therapy ; epidemiology ; virology ; Humans ; Medicine, Chinese Traditional ; Pandemics ; Pneumonia, Viral ; drug therapy ; epidemiology ; virology ; Signal Transduction

Objective To study virulence factors and drug resistance mechanism of linezolid-intermediate Enterococcus faecalis(E.faecalis) isolated from patients with bloodstream infection.Methods Two linezolid-intermediate E.faecalis strains,namely A and B,were isolated from two patients with bloodstream infection,the treatment of two patients was analyzed.The minimum inhibitory concentration (MIC) of linezolid and vancomycin were determined.The virulence genes (esp,asa1,gelE,ace,agg,efaA,cylA,and hyl) and linezolid resistance genes (domain V region of the 23SrRNA,cfr,cfr[B],optrA) were amplified by polymerase chain reaction (PCR).PCR products of domain V region of 23SrRNA gene were sequenced and analyzed.Results Symptoms of two patients who isolated two linezolid-intermediate E.faecalis strains were controlled after accepted linezolid therapy.Strains A and B were both susceptible to vancomycin(MICs were 1μg/mL and 4μg/mL respectively),teicoplain,ampicillin,and nitrofurantoin,while intermediate to linezolid(MIC were both 4μg/mL).Two strains both contained multiple virulence factors,strain A were negative for cylA and hyl,strain B were negative for hyl and esp,but positive for other virulence genes.There was G2621T mutation in domain V region of 23SrRNA in strain A,and no variation was found in strain B.Drug resistance genes of cfr,cfr(B),and optrA were all negative in both strain A and B.Conclusion In the present study,two linezolid-intermediate E.faecalis strains isolated from patients with bloodstream infection were susceptible to vancomycin and ampicillin,although the treatment of linezolid in two patients is effective,the utilization of linezolid therapy in clinical practice still needs to be cautious.The mutation of target site is a significant resistance mechanism,it is necessary for us to pay more attention to these clinical strains which are non-susceptible to such antimicrobial agents,and the treatment strategy needs further study.