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Objective:To investigate the tumor perfusion enhancement induced by low intensity ultrasound stimulated microbubble cavitation (USMC) combined with programmed cell death-Ligand 1(PD-L1) antibody on improving the immune microenvironment of solid tumors.Methods:Tumor-bearing mice were divided into 4 groups: Control ( n=26) group, USMC ( n=27) group, anti-PD-L1 ( n=27) group and USMC+ anti-PD-L1 ( n=27) group. USMC treatment was performed with a VINNO 70 ultrasound theranostics system. Tumor perfusion was evaluated by contrast-enhanced ultrasound (CEUS). The anti-tumor efficacy was assessed by the tumor growth curve and the survival time of mice. The number and function of CD8 + T cells, the differentiation of CD4 + T cells, the proportion of MDSC and the phenotype distribution of TAM in tumors were analyzed by flow cytometry. The content of CXCL9, CXCL10 and HIF-1α in tumor were detected by ELISA. The expression of VEGF in tumor tissues was analyzed by immunofluorescence. Results:CEUS showed that the values of PI and AUC of tumors were significantly increased after USMC compared with before USMC (all P<0.05). USMC combined with anti-PD-L1 therapy did suppress the tumor progression. FCM showed the number, the expression of proliferation antigen Ki67, the secretion of IFN-γ and Granzyme B of CD8 + T cells in tumors were higher in combined group than those in other three groups after therapy (all P<0.05). Meantime, the proportion of Th1 was rose while Tregs and MDSC were declined and the polarization of TAM was toward M1 type by combined therapy. ELISA analysis showed that the combined therapy also increased the concentration of CXCL9, CXCL10 and decreased the content of HIF-1α in tumors (all P<0.05). Meanwhile, the immunofluorescence expression of VEGF was significantly lower in combined group than that in the control group after treatment ( P<0.05). Conclusions:Tumor perfusion enhancement by USMC combined with PD-L1 antibody therapy could improve tumor immune microenvironment and USMC might be a novel effective method for potentiating PD-L1 antibody immunotherapy.
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Objective To study the wear condition of rotating hinge knee prosthesis (RHKP) during horizontal walking gait, and provide technical references for wear prediction and clinical application of this kind of prosthesis. Methods A finite element wear model of RHKP was established based on the standard ISO 14243, and the simulation results including the wear distribution and mass wear rate of tibiofemoral joint surface were compared with the results from in vitro experiments on the same type of prosthesis. Results The mass wear rate of the polyethylene insert was 26.01 mg/MC (million cycle)from finite element analysis (FEA) and (30.06±1.21) mg/MC from in vitro experiments, and the mass wear rate of the upper surface of polyethene insert from FEA was about 3.3 times that of the lower surface. The wear area obtained by FEA was basically consistent with that obtained by in vitro wear measurement. The main wear area was symmetrically distributed in the middle and back of the upper surface. Conclusions The mass wear rate of RHKP, as a semi-restrictive prosthesis, is obviously higher than that of primary total knee joint prosthesis. More attention should be paid to wear test and evaluation of RHKP.
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Objective:To study the effects of different duty cycles [modulated by pulse repetition frequency(PRF)] of diagnostic ultrasound on enhancing blood perfusion in hypovascular tumors.Methods:Walker-256 tumor cells were injected subcutaneously into the inner thigh of 15 SD rats, and the rats were randomly divided into 3 groups according to PRF: group A(PRF=50 Hz), group B(PRF=1.0 kHz) and group C(PRF=2.0 kHz), with 5 rats in each group. The tumor-bearing rats in each group were treated with ultrasound combined with microbubbles for 10 minutes. Contrast-enhanced ultrasound was performed before treatment, immediately after treatment, and 4 hours after treatment, respectively, to compare the peak intensity(PI), area under curve(AUC), and tumor perfusion area in each group.Results:Immediately after treatment, PI and AUC values of group A and B increased compared with those before treatment (all P<0.05); 4 hours after treatment, PI and AUC values of group B continued to increase, superior to that immediately after treatment (all P<0.05), while PI and AUC values of group A and C were decreased compared with those immediately after treatment (all P>0.05). Immediately after treatment, the tumor blood perfusion area in group A and B increased significantly compared with that before treatment (all P<0.05); 4 hours after treatment, the tumor blood flow perfusion area of group B continued to increase, which was better than that immediately after treatment ( P<0.05), and the tumor blood perfusion areas of group A and C decreased compared with that immediately after treatment (all P>0.05). Conclusions:Low-intensity diagnostic ultrasound stimulated microbubble cavitation with PRF=1.0 kHz can enhance tumor blood flow significantly which can last for more than 4 hours.
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Objective: To explore the echocardiographic cardiac geometric morphology and hemodynamics in premature infants at different gestational age with the inlfuencing factors. Methods: A total of 150 premature infants and 150 full-term control infants were enrolled in this study. Based on gestational age, premature infants were divided into 3 groups:①(28-32+6 ) weeks,②(33-34+6 ) weeks,③(35-36+6) weeks; and full term control infants were divided into 2 groups:①’(37-38+6) weeks and②’ (39-41+6) weeks respectively. An iE33 Philips ultrasound examination was conducted to measure left ventricular end-diastolic diameter (LVEDD), LVESD, interventricular septum thickness, posterior wall thickness, left ventricular end-diastolic volume (LVEDV), LVESV, stroke volume, LVEF, left ventricular fractional shortening (LVFS), cardiac output, stroke index, cardiac index, left ventricular mass, left ventricular mass index (LVMI), left ventricular relative wall thickness, left ventricular remodeling index (LVRI) and LVEDVI. Results: With adjusted body surface area, all parameters for cardiac geometric morphology and hemodynamics were similar among different groups,P>0.05. The day-old age (P=0.001), height (P=0.001) and body weight for low weight born infant (P=0.012), for normal weight born infant (P=0.003), for giant infant (P=0.016) were the independent inlfuencing factors for LVMI. The impact of anthropometry and the basic life indexes were similar on LVRI among groups (χ2=42.88,P=0.076), while the covariates were different on LVMI among groups (χ2=123.6,P Conclusion: Cardiac morphology and hemodynamics measured by echocardiography has important clinical meaning for assessing the development and maturity of neonatal hearts in premature infants.
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Objective To clarify whether the NADPH oxidases (NOXs) family contributed to the reactive oxygen species (ROS) production and subsequent interstitial fibrosis in unilateral ureter obstruction (UUO) rats.Methods Male Wistar rats were randomly divided into sham operation group (n =8),sham operation + apocynin treatment group (n =8),UUO operation group (n =8) and UUO operation+apocynin treatment group (n =8).Either vehicle or apocynin (100 mg/kg per day) were given by gavage for 7 days after surgery.Rats were sacrificed at 7th day.ELISA was used to detect the activity of superoxide dismutase (SOD) and catalase (CAT),and the level of 8-iso-prostaglandin F2alpha (8-isoPGF2α) in renal tissue.Western blotting was used to detect the protein expressions of NADPH oxidase subunit NOX2 and NOX4,α-smooth muscle actin(α-SMA),collagen Ⅰ (COL-Ⅰ) and the level of ERK1/ 2 phosphorylation (p-ERK1/2).Results UUO rats with vehicle displayed increased oxidative stress,as measured by renal tissue 8-iso-PGF2α,accompanied with increased renal expression of NADPH oxidases (NOX2,1.5-fold and NOX4,1.7-fold,respectively),compared with sham-operated rats (P ＜0.05).Furthermore,vehicle treated UUO rats showed increased renal COL-Ⅰ and α-SMA levels,compared with sham-operated rats (P ＜ 0.05).ERK1/2 was also activated as detected by p-ERK1/2 expression in UUO rats with vehicle (P ＜ 0.05).Apocynin treatment significantly decreased renal tissue 8-iso-PGF2α level and expressions of NOX2 (-28.7％) and NOX4 (-31.0％) in UUO rats,respectively,compared with vehicle treated rats (P ＜ 0.05).And significant decrease of COL-Ⅰ (-26.4％) and α-SMA expression (-80.0％) were also observed (P ＜ 0.05).The activation of ERK1/2 in UUO rats was greatly inhibited by apocynin treatment (P ＜ 0.01).Despite the pronounced dysregulation of pro-oxidative NOXs family,no compensatory increase of antioxidative enzyme activities occurred.Conclusion The NOXs family contributes largely to the production of ROS and subsequent interstitial fibrosis after ureter ligation,and inhibition of the NOXs family may be a choice for preventing interstitial fibrosis.
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<p><b>OBJECTIVE</b>The imbalance between extracellular matrix (ECM) synthesis and degradation induces the excessive ECM deposition and thus renal fibrosis. The decoction (A&A) which is a combination of two Chinese herbs, Astragalus membranaceus var. mongholicus and Angelica sinensis, has been shown to alleviate ECM production in animal models of chronic kidney diseases. In this paper, the effect of A&A on ECM degradation was investigated with interstitial fibrosis in rats.</p><p><b>METHOD</b>Male Wistar rats were randomly divided into sham, unilateral ureteral obstruction (UUO) and UAA (UUO plus A&A administration) groups. After administration of A&A (14 g x kg(-1) x d(-1)) by gavage for 3, 7 and 10 days, morphological changes were evaluated by HE, PAS and Sirius red staining technique. The expression of plasminogen activator inhibitor-1 (PAI-1) and tissue-type plasminogen activator (t-PA), the activity of PAI-1 and t-PA were determined by ELISA. The activity of matrix metalloproteinases (MMP-9, MMP-2), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) were evaluated by gelatin zymography or reverse gelatin zymography, respectively.</p><p><b>RESULT</b>Morphological analysis showed severe interstitial mononuclear cells infiltration, tubular atrophy, renal fibrosis and collagen expression in kidneys of UUO group, which was reduced by A&A administration (P < 0.05, UAA vs UUO group). Compared with the sham group, the expression of PAI-1 was significantly increased in UUO group by 63%, 91% and 112% at day 3, 7 and 10 respectively; and there were a remarkable decrease in UAA group by 44%, 43% and 52% at day 3, 7 and 10. The expression of active PAI-1 was strikingly increased in UUO group at day 3 [(30.5 +/- 23.8) ng x g(-1) vs. (0.0 +/- 0.0) ng x g(-1), P < 0.05)], day 7 [(36.5 +/- 11.2) ng x g(-1) vs. (0.0 +/- 0.0) ng x g(-1), P < 0.05)], and day 10 [(54.5 +/- 14.2) ng x g(-1) vs. (0.5 +/- 0.5) ng x g(-1), P < 0.05)]. The active PAI-1 was decreased in UAA group at day 7 [(14.9 +/- 0.5) ng x g(-1) vs. (36.5 +/- 11.2) ng x g(-1), P < 0.05] and day 10 [(15.4 +/- 4.8) ng x g(-1) vs. (54.5 +/- 14.2) ng x g(-1), P < 0.05]. The expression of t-PA was increased in UUO group only at day 3 [(58.1 +/- 16.5) microg x g(-1) vs. (30.1 +/- 17.3) microg x g(-1)], P < 0.05), meanwhile decreased in UAA group [(26.3 +/- 8.7) microg x g(-1) vs. (58.1 +/- 16.5) microg x g(-1), P < 0.05)]. But the expression of active t-PA was shown no significantly difference among the three groups. For MMP-2 and MMP-9 activity, they were significantly higher compared with the sham group in UUO group, but no significantly change after A&A treatment. The TIMP-1 activity was significantly increased in UUO group by 28% and 63% at day 7 and 10 respectively, significantly decreased in UAA group by 40% and 39% at the same time point.</p><p><b>CONCLUSION</b>The anti-fibrosis effects of A&A might be associated with modulating the imbalance of PAs/PAIs system as well as MMPs/TIMPs system, thereby alleviate ECM accumulation and interstitial fibrosis.</p>
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Animals , Humans , Male , Rats , Angelica sinensis , Chemistry , Astragalus Plant , Chemistry , Drugs, Chinese Herbal , Extracellular Matrix , Metabolism , Fibrosis , Kidney , Metabolism , Pathology , Kidney Diseases , Drug Therapy , Metabolism , Pathology , Matrix Metalloproteinase 1 , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Plasminogen Activator Inhibitor 1 , Metabolism , Rats, Wistar , Tissue Plasminogen Activator , MetabolismABSTRACT
Objective To investigate the influence of a combination of Chinese herbs,Astragalus membranaceus var.mongholicus and A ngelica sinensis (A&A)on the production and elimination of reactive oxygen species (ROS)and the underlying mechanism during the process of renal interstitial fibrosis in the obstructive kidneys of rats with unilateral ureteral obstruction(uuo).Method Male Wistar rats were randomly divided into control,sham,UUO and UAA (UUO+A&A)groups.The rats in UAA group were administered with A&A(14 g/kg)by oral gavage once daily:the ones in sham and UUO group were given with equal volumes of water.Three days after setting up models,pathological injury of renal tissue was evaluated.Level of total antioxidant capacity(T-AOC)and activity of CuZn superoxide dismutase(CuZn-SOD)in renal homogenates were measured bv spectrophotometry.Expression of NADPH oxidase subunits p47-phox,p22-phox and nintrotyrosine was analyzed by Western blot. Results Severe interstitial inflammatory cells infiltration, mild tubular atrophy and interstitial fibrosis were found in UUO group, which were alleviated by A&A administration. Compared to sham group, T-AOC of UUO group was not significantly changed, but the expression of the nitrotyrosine, NADPH oxidase subunits p47-phox and p22-phox was increased significantly (P<0.05). After A&A administration, T-AOC level was increased (2.5±1.1 vs 1.5±0.5, P<0.05) and the expression of nitrotyrosine was decreased (P< 0.05) in UAA group compared with UUO group. Additionally, the expression of p47-phox was reduced at day 3 (P<0.05), accompanied with a reduced expression of p22-phox (P< 0.05). CuZn-SOD activity was not significantly changed among the groups. Conclusion The inhibition of A&A in NADPH oxidase subunits p47-phox and p22-phox, which is responsible for reduction of oxidative stress, is associated with the alleviation of renal fibrosis in obstructive rat kidney.
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Objective To investigate whether the activation of secretory prophospholipase A2 (sPLA2) plays the role in the pathophysiological mechanism of acute aristolochic acid nephropathy (AAN) in rats. Methods Wistar rats were randomly divided into two groups. Model group received decocted Aristolochia Manshuriensis Kom 30 g·kg-1d-1 by gavage for 7 days following tap water in same way for additional 7 days. Control group received only tap water by garage at parallel time. The renal pathological changes were observed at the 4th, 8th and 14th day. The injury of renal tubules and interstitium was observed under light microscope following a semi-quantity grade. The level of Scr was measured to evaluate glomerular function. Urinary N-acetyl-beta-glucosaminidase (NAG) was tested as renal tubular injury marker. The activity of sPLA2 in serum was detected by manifesting the color of thiols in the substrate. The protein expression of renal cortex and medulla COX-2 was analyzed by Western blot. The metabolic products of pretaglandins (PC, s) including 6-kcto-PGF1α and TXB2 in the plasma and urine were assayed by radioimmunoassay. The ratio of 6-keto-PGF1α/TXB2 was calculated. Results After Aristolochia administration, the tubulointerstitial injury and Scr increased in AA rats and reached the peak at the 8th day, the tubulointerstitial injury index(8.14±2.55 vs 1.50±0.71, P<0.05) and Scr[(0.24±0.10) vs (0.19±0.02) μmol/g, P<0.05] increased significantly in AA rats compared with control group. The activity of sPLA2 (μmol ·min-1·mg-1) in AA group elevated by 1.3-fold compared to control group at 8th day (133.15±17.05 vs 101.3±16.07, P<0.05), while theexpression of COX-2 in renal cortex increased (1.16±0.36 vs 0.69±0.28, P<0.05) with no change in renal medulla. Even though the levels of serum 6-keto-PGF1α and TXB2 did not change obviously in both AA and control group, but urinary levels of 6-keto-PGF1α and TXB2 increased by 2-fold and 3-fold in AA group compared to control group, respectively (all P<0.05), while the ratio of 6-keto-PGF1α/TXB2 decreased significantly (207.53±17.52 vs 296.64±51.31, P<0.05). All of above changes recovered to the control level at the 14th day except the tubulointerstitial injury index. Conclusion Serum sPLA2 is activated in the rots with acute kidney injury induced by aristolochic acid, which accompanied by up-regulated expression of COX-2 in renal cotex and increased the metabolic products of vasoconstrictive PG s in urine. These changes may participate the mechanism of renal peritubular ischemia in AAN.
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0.05).AA-I could induce the TGF-?1 secretion by HK-2 cells(18.26?5.98 ?g/L,vs.control,P
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Objective To explore the relationship between diethylhexylphthalate (DEHP) and peritoneal sclerosis and its possible mechanism. Methods Isolated human peritoneal mesothelial cells (HPMC) was cultured for five days in culture solution with varied doses of DEHP ７．７?of 10-6mol/L (A group), 5.4?１０－５mol/L (B group) and 2. 2?10-4 mol/L (C group) respectively. Culture solution without DEHP was as control. Total collagen synthesis and secretion was assayed by 3H-proline incorporation. ELISA method was used to detect the level of secreted fibronectin (FN) . The expression of FN mRNA , collagen ImRNA, collagen Ⅲ mRNA and TGF－?l mRNA was determined by RT-PCR. Results (1)HPMC expressed FN, laminin, collagen Ⅲ and TGF-?１. (2)DEHP stimulated FN and collagen expression with a higher level in C group compared with control(P