Objective To develop and validate a ultrasonographic (US)imaging agent with targeted microbubbles that attaches to chemokine receptor 2 (CCR2)and to compare the US single obtained from targeted microbubble with that from control microbubble in murine breast tumor model.Methods The microbubble which carried CCR2 antibody (MBCCR2 )and isotype-macthed immunoglobulin G-labled control microbubble (MBcontrol ) were prepared. The microbubble size and distribution were assessed by AccuSizer780.Binding specificities of targeted microbubble compared with control microbubble were tested with murine microvascular endothelial cells (bEnd.3 ).Orthotopic breast tumor model was estabished in BALB/c mice with mouse breast cancer 4T1 cell.In vivo imaging signals of contrast material-enhanced ultrasound by use these two different types of microbubble which were injected respectively into each mouse at random order and 30 min interval.Tumor tissue was stained for CCR2 and CD3 1 .Results Automatic Particle Sizer showed size uniform of two kinds of microbubbles,and narrow distribution of particle size (mean diameter of about 1 -2 μm),which were not significantly different (P >0.05).Adhension to bEnd. 3 endothelial cells was significantly higher (P < 0.001 )for MBCCR2 (mean,9.50 ± 1 .5 1 )than that for MBcontrol (mean,0.01 ±0.01).Imaging signal in the murine tumor model was significantly higher for MBCCR2 [mean,(6.76±0.26)dB]than that for MBcontrol [mean,(1 .06 ±0.62)dB,P <0.001 ].Immunofluorescence confirmed expression of CCR2 on tumor vasculature.Conclusions The targeted microbubbles with CCR2 monoclonal antibody had been successfully prepared,which precisely targeted to CCR2 of tumor angiogenesis in the murine breast cancer xenograft tumor models in vivo.These results suggest that the targeted microbubbles as a kind of ultrasound molecular imaging agent with a better specificity can be used for both evaluating tumor neovascularization and monitoring therapeutic effect of anti-angiogenesis.

The effect of triptolide on proliferation and apoptosis of human multiple myeloma RPMI-8226 cells in vitro, as well as the roles of nuclear factor-kappa B (NF-κB) and IκBα was investigated. The effect of tritptolide on the growth of RPMI-8226 cells was studied by MTT assay. Apoptosis was detected by Hoechest 33258 staining and Annexin V/PI double staining assay. The expression of NF-κB and IκBα was observed by Western blot and confocal microscopy. The results showed that triptolide inactivated NF-κB apoptotic pathway in human multiple myeloma RPMI-8226 cells. Triptolide at nM range induced proliferation inhibition in a dose- and time-dependent manner and apoptosis in a dose-dependent fashion in RPMI-8226 cells. Besides, we observed the inhibition of NF-κB /p65 in the nuclear fraction was correlated with the increase in the protein expression of IκBα in the cytosol. These results suggested that triptolide might exhibit its strong anti-tumor effects via inactivation of NF-κB/p65 and IκBα.

Objective To prepare Integrin αvβ3/CCR2 dual targeted microbubble,test physical-chemical properties,enhancement effect and targeting ability in vitro.Methods The dual targeted microbubbles (MBdual-target )with FITC labled iRGD and PE labled CCR2 were prapared,and non-target microbubbles as control (MBcontrol ) were prapered.Physical and chemical properties of two groups of microbubbles were tested,connectivity of peptides/antibodies and microbubbles were detected by fluorescence microscope and flow cytometry instrument.Enhancement effect and the stability of two groups of microbubbles was observed and compared in vitro.The affinity of MBdual-target and MBcontrol for bEnd.3 cells was investigated with light and fluorescent microscope.Results ①The particle size of MBdual-target was (0.93±0.23)μm,with no statistically significant difference compared with MBcontrol (P >0.05).②MBcontrol showed no fluorescent,while MBdual-target showed both clear green and red light,under fluorescent microscope.③There was no significant difference of gray scale of enhancement between MBdual-target and MBcontrol in vitro.④ It was showed that MBdual-target adhered to bEnd.3 cells in vitro experiment.Conclusions Integrinαvβ3/CCR2 dual targeted targeted microbubbles was successfully prepared and proved having good enhancement effect and targeting ability in vitro.

Objective To evaluate the short term results of repairing osteonecrosis of the femoral head by modified free vascularized fibular grafting through Smith-Peterson approach without cutting the rectus femoris.Methods Between January,2009 and September,2012,14 osteonecrotic hips of 14 patients who underwent free vascularized fibular grafting were retrospectively studied including 10 male and 4 female.The follow up period was 24 months.The preoperative and postoperative symptoms relief,recovery of hip function (Harris score),radiographic assessment (X-ray).Results All patients got postoperative ipsilateral hip pain relief.Average Harris hip score improved significantly (68.47 ± 5.33,86.80 ± 4.72 points in preoperation and postoperation,respectively; paired t test,t =18.724,P ＜ 0.01).Postoperative X-ray results showed there were no progresses of osteonecrosis and no collapse in the operative side within 2 years follow-up.Conclusion The short term treatment results of femoral head avascular necrosis were satisfactory by using free vascularized fibular graft through Smith-Peterson approach without cutting off the rectus femoris and it may be a relatively practical option of surgical approach.

Objective To study the correlation between the cytochrome P450 3A5 gene polymorphism and essential hypertension (EH) in Chinese population .Methods The real-time PCR genotyping at CYP3A5*3(6986A>G) position was established using Taqman minor groove binding (MGB) probes .Total 170 EH patients and 193 matched controls of Chinese Han population were genotyped at CYP3A5*3(6986A>G) position using this method .Results The GG ,GA ,AA genotyped frequencies were 51 .2% , 42 .4% and 6 .5% for the EH patients and 39 .9% ,50 .8% and 9 .3% for the control group respectively .The risk of EH for person carrying GG genotype was 1 .579 fold of the persons carrying at least one A allele(95% CI:1 .041-2 .395) .Conclusion CYP3A5*3(6986A>G) polymorphism may be associated with EH in Chinese population .The risk of EH is decreased in the persons carrying allele A ,slightly lower levels of systolic blood pressure exists .

Objective To compare the advantages and disadvantages of three different fixation methods via muscle-splitting approach for the treatment of single segmental lumbar vertebral diseases.Methods All of 90 patients with the single segmental lumbar vertebral diseases were treated from June 2012 to June 2013,including 34 males and 56 females,at the age of 27-76 years old,averagely (53.37± 15.41) years.Twenty-seven patients were treated with unilateral pedicle screws via unilateral muscle-splitting approach (unilateral fixation group).Thirty-three patients were treated with unilateral pedicle screws and contralateral translaminar facet screw via unilateral muscle-splitting approach (combined fixation group).Thirty patients were treated with unilateral pedicle screws via bilateral muscle-splitting approach (bilateral fixation group).Observe and compare the imaging and clinical results of these three groups.Results There were no significant difference in incision length and postoperative wound drainage between these three groups.The operation time was shortest in unilateral fixation group,while longest in bilateral fixation group.In the intraoperative blood loss,the unilateral fixation group was the same as the combined fixation group,while less than the bilateral fixation group.No infection was found after operations.Seven patients experienced epidermal necrosis of the incision,including 1 patient of unilateral fixation group,4 patients of combined fixation group and 2 patients of bilateral fixation group.Three patients suffered from nerve injury,including 2 patients of combined fixation group and 1 patients of bilateral fixation group.The follow-up time was from 12 to 36 months,averagely 25.5 months.The intervertebral height of lesion segments was apparently recovered after surgery,and maintained during the follow-up in these three groups.No significant changes of area and grade of multifidus muscle were detected at 1 year after surgery compared with preoperative.Seven patients experienced endplate cutting and cage partially embedded into vertebral body,including 3 patients of unilateral fixation group,3 patients of combined fixation group and 1 patient of bilateral fixation group.There was no loosening,displacement,breakage or cage shifting.Except 5 patients (2 patients of unilateral fixation group,2 patients of combined fixation group and 1 patient of bilateral fixation group),the remaining cases had well interbody fusion.At the final follow-up time,the JOA score significantly increased compared with preoperative.Besides,unilateral fixation group and combined fixation group were superior to bilateral fixation group.Conclusion Three different fixation methods via muscle-splitting approach for the treatment of single segmental lumbar vertebral diseases had well clinical results.The unilateral fixation and combined fixation via muscle-splitting approach had more advantages.The combined fixation via muscle-splitting approach is an innovation in approach,exposure and fixation aspects.

Aim To study on the mitochondrial mass change in mouse thymocyte apoptosis.Method Control and Dexamethasone(DEX) groups were set;at 6h,we studied mitochondrial mass changes by NAO and Mitotracker Green(MG) staining flowcytometry and detected mitochondrial membrane potential change with DiOC_6(3) staining flowcytometry.We also used Annexin V-PE/MG double staining flocytometry to examine the mitochondrial changes in apoptosis progress.Results NAO staining results showed that 1 ?mol?L~(-1) DEX stimulation reduced the cardiolipin content of thymocyte mitochondria(P

In this study,nearly 200 endophytic bacteria were isolated from different part of Huperzia serrata, over 60 bacterium with clear antifungal activity were selected from those cultures.Among them,strain H-6 exhibited the highest antifungal activity which was strongly inhibits the growth of many plant pathogenic fungi such as Sclerotinia scleroliorum,Fusarium graminearumt,Sclerotinia libertiana,Phytophthora capsici Leonia and Sesame fusarium wilt.According to the characteristics of morphology,physiology and biochem- istry tests and the comparison of 16S rDNA sequence,the strain H-6 was similar to the Burkholderia.So strain H-6 was identified as Burkholderia sp.H-6.The results also showed that Burkholderia sp.H-6 was markedly different from Burkholderia cepacia that was applied widely in agriculture as antagonistic bacteria. The medium and culture conditions of the strain all were optimized by single factor and orthogonal experiments.In the investigation of the culture condition,growth was carried out in a basal medium(potato juice)and gradually supplemented with the various ingredients to be investigated.The major ingredients be- ing investigated included carbon sources and nitrogen sources.The optimal antifungal activity production condition is growth in a medium(potato juice with 2.5%mannitol and 0.1%NaNO3),initial pH 4.0 at 28℃.

OBJECTIVETo study the pathogenesis of adrenal mitochondrial dysfunction in septic rats.

METHODSThirty SPF rats were randomized into 3 groups, including a normal control group and 2 sepsis groups receiving intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS) and observed at 6 and 24 h after the injection. The adrenal mitochondria were extracted from the rats at the corresponding time points for observation by electronic microscopy. The membrane potential of the mitochondria was detected by flow cytometry. The oxidative stress levels in the mitochondria (activities of NOS and levels of MDA and NO) were assessed.

RESULTSWith the progression of sepsis, the serum levels of corticosterone in LPS groups increased significantly as compared with that in the control group. Ultrastructural observation of the adrenal mitochondria showed mild mitochondrial injury in LPS groups in comparison with the control group. The mitochondrial membrane potential was lowered in the LPS groups, but all these changes appeared to be reversible. The activities of NOS and the levels of MDA and NO showed no significant difference between the sepsis groups and the control group.

CONCLUSIONSNo obvious adrenal dysfunction occurs in the early stage of sepsis in rats. Mitochondrial injury, which is reversible, occurs in early sepsis without obvious evidence of oxidative stress injury in the adrenal mitochondria, suggesting a strong resistance and capacity of self-repair of the adrenal gland and the mitochondria against sepsis-induced injury.

Adrenal Glands ; metabolism ; Animals ; Disease Models, Animal ; Male ; Membrane Potential, Mitochondrial ; Mitochondria ; metabolism ; pathology ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Sepsis ; metabolism ; pathology

OBJECTIVETo compare the changes of inward rectifying K(+) (Kir) current (I(K1)) density in atrial myocytes of patients with rheumatic atrial fibrillation (RAF) less or longer than 6 months.

METHODI(K1) density was measured with whole-cell patch clamp technique in single myocyte isolated by an enzymatic dissociation method from right atrial appendages in patients with RAF less than 6 months (n = 18) and longer than 6 months (n = 18), RAF patients with normal sinus rhythm (NSR, n = 18) served as control.

RESULTSThe average resting membrane potentials were similar between various groups. I(K1) density in single myocyte at -50 to -100 mV of patients with RAF longer than 6 months was significantly increased compared to that in patients with RAF less than 6 months and NSR patients. I(K1) density in single myocyte at hyperpolarized potential level (-100 mV) was also significantly increased in patients with RAF longer than 6 months (8.94 +/- 0.15) than that in patients with RAF less than 6 months (4.35 +/- 0.49) and NSR patients compared to that in NSR (4.05 +/- 0.96, P < 0.05 vs RAF longer than 6 months).

CONCLUSIONThe data suggest I(K1) current increase might contribute to the electrical remodeling in RAF patients.

Adult ; Atrial Fibrillation ; metabolism ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Myocytes, Cardiac ; metabolism ; Patch-Clamp Techniques ; Potassium ; metabolism ; Potassium Channels, Inwardly Rectifying ; metabolism