Objective To investigate the effects of leptin on Th17 cells and the possible mechanism. Methods The leptin-deficient ( ob/ob) mice and their homologous wild-type mice were used in the study.The percentages of Th17 cells in peripheral blood samples, spleen tissues and lymph nodes were measured by flow cytometry ( FCM) analysis.The splenic CD4+T cells, separated from the ob/ob mice and the wild-type mice by using magnetic beads,were respectively cultured with leptin at various concentrations and with anti-leptin neu-tralization antibody to evaluate the effects of leptin on Th17 cells.The quantitative real-time PCR was performed to analyze Th17 cell-related cytokines at transcriptional levels.The levels of IL-6 and IL-17A in the supernatants of CD4+T cell culture were measured with Luminex technology.Results Compared with the wild-type mice, the ob/ob mice showed lower percentages of Th17 cells in both peripheral blood samples and spleen tissues (0.49%±0.03%vs 1.29%±0.1%, 1.56%±0.22%vs 2.47%±0.11%).There was a decrease in the percentages of Th17 cells upon the in vitro treatment of CD4+T cells from wild-type mice with anti-leptin antibody.The per-centages of Th17 cells were increased in a dose-dependent manner upon the in vitro treatment of CD4+T cells from ob/ob mice with leptin.Moreover, the levels of IL-17A and IL-6 and the transcriptional levels of RORγt, IL-17A and IL-6 in leptin deficiency group were lower than those of wild-type group, but were increased upon the treatment with leptin.No significant difference with the transcriptional levels of TGF-βand IL-23 was ob-served between the two groups with and without intervention.Conclusion Leptin deficiency seriously hampered the generation of Th17 cells in mice and resulted in a decreased expression of RORγt, IL-17A and IL-6 at mRNA level.The treatment of CD4 T cells with leptin might promote the generation of Th17 cells through up-regulating the transcription of RORγt and IL-6.

Objective:To explore the correlation between the characteristics of contrast-enhanced sonography of intraoperative glioblastoma multiform (GBM) and molecular markers of isocitrate dehydrogenase-1(IDH1).Methods:A retrospective analysis were performed in 30 patients who underwent neurosurgery and pathologically confirmed to be GBM at Beijing Tiantan Hospital from May 2018 to April 2019. All neurosurgical glioblastoma patients after craniotomy underwent conventional ultrasound and contrast-enhanced ultrasound(CEUS) guided navigation. The characteristics of the ultrasound imaging (whether the tumor involves the structure of the corpus callosum, the clarity of the tumor boundary after enhanced ultrasound and whether the tumor has necrotic areas with enhanced ultrasound images) were analyzed. The ratio between tumor necrosis area and whole tumor area (N/W) was measured, and the correlation with IDH1 gene expression was analyzed.Results:There were statistical differences in clarity of tumor boundary after CEUS and tumor necrosis after CEUS between positive IDH1 and negative IDH1 groups(all P<0.05). The positive expression of IDH1 was negatively correlated with the N/W area of the contrast-enhanced ultrasound mode( r=-0.756, P<0.05), suggesting that the expression level of IDH1 gene was negatively correlated with the area of tumor necrosis. Conclusions:Ultrasound contrast agent examination can more accurately distinguish the active proliferation area, hemorrhagic necrosis area and peripheral edema area of glioblastoma. Accurately identifying the extent of tumor necrosis area through ultrasound contrast agent examination can predict expression of IDH1.

Objective To observe the influence of trastuzumab on DNA break repair and Her-2 nuclear import after radiation in breast cancer cell line SKBR3,and discuss the radiosensitivity mechanism of trastuzumab.Methods Clone formation assay was used to analyze the difference of survival fractions between radiation group and radiation plus trastuzumab group.Confocal microscopy was applied to observe the influence of trastuzumab in the nuclear import process of Her-2 and the expression of γH2AX after radiation,which is considered as the marker of DNA double strand break.Western blotting was used to detect the expression of Her-2 and DNA-PKcs in nuclei after radiation.Results The result of clone formation assayshowed that the SF2 in radiation group was 0.547±0.046 and 0.321±0.022 in the radiation plus trastuzumab group were significantly decreased,the results of confocal microscopy showed that trastuzumab postponed the nuclear import process of Her-2 (52.80±19.74 in radiation group,21.41±10.55 in the radiation group),and increased expression of γH2AX after radiation (85.40±25.63 in radiation group,18.53±44.32 in the radiation group),and western blotting revealed trastuzumab reduced the expression of Her-2,DNA-PKcs in nuclei.Conclusion Trastuzumab can inhibit the radiation induced nuclear import of Her-2,and decrease Her-2,DNA-PKcs in nuclei to increase the DSB on early stage after radiation.

ObjectiveToexplore the appearance ofmusclecrushinjuryatquantitative ultrasonographic elastography by supersonic shear imaging (SSI).MethodsThe animal experiment was done using a special balloon cuff device to create left hind leg crush injury with a force of 18.6 kPa.Twentythree New Zealand rabbits had crush injury of extremity and survived for 72 hours.SSI quantitative elastography was performed in crushed and no-crushed regions of each rabbit hind leg.Quantitative lesion elasticity was measured in terms of the Young modulus (in kilopascals) at 30 min,2 h,6 h,24 h and 72 h after the release of the crushing pressure.A receiver operating characteristic (ROC) curve analysis was used to assess diagnostic performance.ResultsThe area of crushed region in left hind leg accounted for 2.6％ -3.0％ of body surface area in 23 rabbits.The crushed regions exhibited maximum elasticity values of (19.51 ± 6.74)kPa,(21.47 ± 5.54) Pa,(11.36 ± 5.35)kPa,(15.09 ± 3.31)kPa and (13.72 ± 3.74) kPa,and mean elasticity values of (12.44 ± 3.77)kPa,(13.20 ± 3.60)kPa,(6.80±2.86)kPa,(10.04 ± 2.95)kPa and (6.94 ± 0.97)kPa at 30 min,2 h,6 h,24 h and 72 h after the release of the crushing pressure.Comparing with those of no-crushed regions,they were higher obviously (P＜0.001).ROC curves showed that extremity crush injury was diagnosed by using elasticity value,and the greater the elasticity value,the greater the diagnostic value.Conclusions SSI provides quantitative elasticity measurements,thus adding complementary information that potentially could help in crush injury characterization with conventional ultrasonography.

This study examined the radiation-induced ERBB2 nuclear transport in the BT474 breast cancer cell line and the relationship between caveolin-1 and radiation-induced ERBB2 nuclear transport. The BT474 cells were treated with herceptin (200 nmol/L), PP2 (a caveolin-1 inhibitor, 100 nmol/L) and irradiation combined or alone. Confocal microscopy was used to observe the nuclear import of ERBB2 and caveolin-1 after irradiation. Western blotting was employed to detect the expression of ERBB2, caveolin-1 and DNA-PKcs after irradiation, and immunoprecipitation to identify the ERBB2 and caveolin-1 complex before perinuclear ERBB2 localization. Confocal microscopy showed the transport of ERBB2 and caveolin-1 from the cell membrane to the nucleus 15 min after irradiation and the proteins accumulated at the perinuclear region within 45 min. Western blotting revealed that the expression levels of ERBB2, caveolin-1 and DNA-PKcs were increased after irradiation and reached a peak 45 min later. Both herceptin and PP2 treatments were found to decrease ERBB2 expression. An immune complex composed of ERBB2 and caveolin-1 was found in the herceptin group after irradiation. It was concluded that after irradiation, ERBB2 may be transported from the cell membrane to the nucleus and activate DNA-PKcs to trigger DNA double-strand break (DSB) repair; caveolin-1 may participate in this process. Treatments involving the downregulation of caveolin-1 may increase the radiosensitization of breast cancer cells.

Objective To approach the diagnostic value of contrast-enhanced ultrasonography(CEUS)for the detection of traumatic laceration of pancreas. Methods Sixty cases of pancreatic traumatic model were made in twelve healthy swines after the animals were anesthetized and laparotomized. Then the conventional ultrasonography(US) and CEUS were performed in each case to diagnose the traumatic region,immediately. The results were compared with surgical findings. Results Among sixty injuries,the detection rate of conventional ultrasonography was 66. 7%,the detection rate of CEUS was 88.3%. Conclusions CEUS shows higher detection rate than conventional US in diagnosing pancreatic laceration,and it also can improve the diagnostic value of ultrasound for the detection of pancreatic laceration.

Objective To study feasibility of combined haemostatic percutaneous injection therapy guided by contrast-enhanced ultrasonography (CEUS) in treatment of renal injuries. Methods Eighteen New Zealand rabbits were inflicted with kidney injury imitating grades Ⅲ-Ⅳ blunt injuries. The animals were randomly and equally divided into three groups, Group A ( treated with hemocoagulase),Group B ( treated with hemocoagulase and Alpha-cyanoacrylate) and Group C ( control group, given normal saline). The hemostatic time, hemostatic effect, and perirenal hematoma were observed. Results A perirenal hematoma was observed one hour after treatment. The perirenal fluid thickness was (0.200 ±0.012) cm in Group A, (0.050 ±0.002) cm in Group B and (0.400 ±0.009) cm in Group C, with statistical significance between two test groups and Group C (P ＜ 0.05 ). At days 7 and 14 following treatment, lesion length and cross section was ( 1. 107 ±0. 143) cm and (0.433 ±0. 163) cm in Group A, (0.567 ±0.082) cm and (0. 160 ±0. 078) cm in Group B, and (0.980 ±0. 203) cm and (0.686 ± 0. 157) cm in Group C. There was statistical significance between the test groups (Groups A and B) and Group C (P＜0. 01) at day 14. The lesion size in Group A was lager than that in Group B (P ＜ 0.01 ). One month after treatment, a slight nephrohydrosis occurred in Group B. Conclusions Either injection of simple hemocoagulase or combined use of hemocoagulase and Alpha-cyanoacrylate guided by CEUS can attain positive hemostatic effect, but the latter one is more rapid and reliable.

Objective To study haemostatic percutaneous injection therapy for the management of vascular damage in patients with renal injuries guided by contrast-enhanced ultrasonography(CEUS).Methods Which of 56 patients with renal trauma were diagnosis by CEUS,37 cases with grades Ⅱ-Ⅳ renal injuries were brought into our study.According to wound degree and accompanying active bleeding,they were divided into experiment group (percutaneous injection hemostatic treatment)and control group(conservative treatment).Results Thirty-seven renal trauma manifest low perfusion in lesions by CEUS,and the contrast agent could be seen overflow to renal pelvi and the location of capsule in 13 patients.The patients were divided into experiment group(17 cases)and control group(20 cases).The color of hematuria of 9 patients in experiment group became gradually light at 30 mins after treatment.and the color of 7 cases become normal,and hematuria of the only one was iterative appear.The color of hematuria of 9 patients in control group became gradually light in 24-72 hours,others' hematuria became gradually light in 5-14 days.The time of color of hematuria become light of the former was shorter than those of the latter(P<0.05).Reexamination by ultrasound and renal function and urine routine at 1,3 and 6 months after treatment,the results of all patients indicated normal.Conclusions Haemostatic percutaneous injection therapy for renal trauma guided by contrast-enhanced ultrasonography has very obvious hemostatic efficacy.Its advantages included may be used for effective,minimally invasive control of renal injuries(grades Ⅱ-Ⅳ),and can be a feasible management of active bleeding at bedside.

Aim To investigate the effect of polysac-charide of Cistanche deserticola ( CDPS) on the impro-ving ability of synaptic plasticity in memory acquisition impairment model mice induced by scopolamine. Methods The KM mice were randomly divided into six groups:scopolamine group, control group, CDPS-treated (25, 50, 100 mg·kg-1 ) group and donepezil group. Memory acquisition impairment model in mice was established with i. p. scopolamine (4 mg·kg-1 ) only once, and orally administered CDPS (25, 50, or 100 mg · kg-1 ) daily for 6 weeks before scopolamine injection. Experimental groups were subjected to step-down test and Morris water maze test. Western blot and RT-PCR analysis were used to examine the expression of GAP-43 , SYP and PSD-95 . Transmission electron
microscope was used to observe the change of synaptic number and structures. Results CDPS (25,50,100 mg·kg-1 ) could shorten the incubation period of mice in the water maze test. Control group and CDPS-treated group swam longer in Q3 than scopolamine group. Mo-reover, CDPS (50,100 mg·kg-1 ) could significantly reduce the error times and extend the incubation period in the step-down test. The results of Western blot and RT-PCR showed that CDPS significantly improved the expression of GAP-43 at the dose of 25 ,50 mg · kg-1 and SYP at the dose of 25,50, 100 mg·kg-1 in hip-pocampus of mice. However, the biochemical assays did not reveal a significant difference in the basal hipp-ocampal levels of the PSD-95 . The ultra-thin speci-mens of hippocampus showed that the number of syn-
apse was increased in CDPS-treated group. Conclu-sions Scopolamine can induce the learning and mem-ory deficits in mice to make related protein expression abnormalities in hippocampus mice, thus this causes the change of synaptic plasticity, which leads to a change in the ability of learning and memory. And CDPS can improve the expression of SYP and GAP-43 ,
increase number of synapses, recover synaptic plastici-ty, and improve the ability of learning and memory in mice.

Objective:To explore a new method of the cultivation of adoptive immunotherapy cells.Methods: Mononuclear cells was isoplated by density gradient centrifugation and then proliferated by using CD 40-agonist monoclonal antibody 5C11、cytokine of IFN-αand IL-7(CD40 group)in vitro.During the culturing procedure ,the cell morphology was obersved by optical microscope.The percentage of T-lymphocytes, NK-T cells, Treg cells and the cell proliferation, which were compared with CIK group CD3mAb activated,was detected on the 9th day.Results:There was no significant difference of CD 4+/CD8+T cells percentage between the two groups.But the Treg cells percentage of CIK group was far higher than that of CD 40 group,while the percentage of CD3+CD56+NK-T cells was lower than that of CD 40 group.And a group of Mo-NK-DC cells were observed in the CD 40 group.Conclusion: The new method of adoptive immunity therapy has been established in this study could increase the percentage of NK -T cells which had the ability to kill tumor cells.Simultaneously ,it is reduced the amount of Treg cells significantly.