BACKGROUND: In recent years, ultrasound microbubble gene transfer system has been applied for gene transfection in many parts of the body, but it has been seldom reported to be used for gene transfection in bone parts. OBJECTIVE: To investigate the efficiency and feasibility of ultrasound-targeted microbubble destruction applied for transfection of enhanced green fluorescent protein plasmid into the femoral head of rabbits.METHODS: Japanese big-ear rabbits were randomly divided into five groups: bare transfection, pre-irradiation + bare transfection, ultrasound transfection, pre-irradiation+ultrasound transfection, and repeatable transfection. In the first two groups, ultrasound-targeted gene transfection and irradiation was not used, but in the latter three groups, ultrasound-targeted microbubble destruction was used to transfect enhanced green fluorescent protein (EGFP) plasmid into the femoral head of rabbits. At 1 week after transfection, EGFP expression in femoral head was observed under the fluorescence microscope. RESULTS AND CONCLUSION: EGFP expression appeared in the ultrasound transfection, pre-irradiation + ultrasound transfection and repeatable transfection. The transfection efficiency of EGFP plasmid was significantly higher in the repeatable transfection group than in the other groups (P < 0.01). Obvious injury loci were not observed in the soft tissue and bone tissue slices of ultrasonic irradiation parts in the ultrasound transfection, pre-irradiation + ultrasound transfection and repeatable transfection groups. These results confirm that ultrasound-targeted microbubble destruction is a safe and effective method to transfect EGFP plasmid into the femoral head of rabbits.

Objective To examine the association of employment-related events with depression and anxiety in senior college students in China.Methods Altogether 1321 senior college students were recruited from Central South University and the survey was administered anonymously. Anxiety and depression were assessed with the Self-rating Anxiety Scale (SAS) and the Center for Epidemiologic Studies Depression Scale (CES-D), respectively. Employment-related events were reflected by the employment-related options when students were surveyed. Logistic regression was used to estimate the effect of employment-related events on anxiety and depression.Results We collected 1178 valid questionnaires. Taking 50 and 16 as the cutting-point of SAS and CES-D, we found 18.9% and 55.4% of the participants showed symptoms of anxiety and depression, respectively. After controlling confounding factors, those students who had taken the entrance exam of graduate schools and were optimistic for the result, those who had taken the entrance exam of graduate schools but were pessimistic for the result, and those who neither took the entrance exam of graduate schools nor received a job offer, had a higher risk of anxiety than those who would attend a graduate school without exam, and the odds ratios were 2.5, 2.3, and 3.5, respectively. For students' depression, the odds ratios of the three categories mentioned above were 2.0, 1.5, and 2.6, respectively. Conclusion Senior college students who face bad employment-related events are at a higher risk of anxiety and depression.

Objective To evaluate the correlation between thin-section CT morphologic features and pathological vascular invasion in the clinical stage ⅠA peripheral lung adenocarcinoma.Methods The preoperative thin-section CT images,clinical and pathological characteristics of 102 nodules in 98 patients were retrospectively analyzed and divided into two groups according to pathological vascular invasion.Variables including the diameter of tumor(T) and consolidation part (C) in the mix ground glass nodule (mGGN),C/T ratio,morphology and pathological classification were analyzed by two-independent samples Mann-Whitney U test or t test and x2 test or Fisher exact probability test.Variables with significant difference in the univariate analyses were entered into multivariate analysis to explore predictors for vascular invasion.The correlation between these measurements and vascular invasion was evaluated by a ROC analysis.Results All of 102 nodules,36 had pathological vascular invasion and 66 without.The numbers of two groups with speculation,air bronchogram,abnormal vein,vascular convergence,pleural indentation were 30,30;26,26;35,3;35,39;32,39 and the mediandiameter of tumors were 2.1(1.0-3.0),1.2(0.5-3.0) cm respectively.By univariateanalysis,spiculation,air bronchogram,abnormal vein,vascular convergence,pleural indentation,and diameter of tumor were significantly associated with pathological vascularinvasion (P＜0.01).By multivariate logistic analysis,the statistically significant difference were found in speculation and abnormal vein (OR=33.867,8 161.063,P＜0.05).The ROC analysis showed the area under curve,cut-off value,sensitivity and specificity ofthe diameter of tumor and consolidation part of mGGN and C/T ratio were 0.784,2.05 cm,66.7％,78.8％;0.886,1.08 cm,82.4％,90.0％ and 0.861,0.65,79.4％,85.0％ respectively in differentiating tumors with pathological vascular invasion.Conclusion The spiculation sign and abnormal vein are independent predictors for vascular invasion in stage ⅠA peripheral lung adenocarcinoma.

This study was designed to investigate the impact of ultra-filtration extract mixture from Astragals mongholicus (UEMAM) o radiosensitivity of H22 ascitic tumor in mice to 12C6+ ions radiation. The H22 ascitic tumor model was established in mice by intraperitoneal injection of 0.2 mL H22 ascitic cells. The animals were subsequently divided into 4 groups randomly, treated with normal saline, UEMAM, heavy ion beam radiotherapy and UEMAM plus heavy ion beam radiotherapy, respectively. The body weights, abdomen circumference of the mice were measured and the mouse behavior was monitored every day; survival time was recorded to evaluate life extension effect; flow cytometry technique was used to detect H22 cell apoptosis and cell cycle; protein levels of p53, Bax, Bcl-2 and cleaved Caspase-3 were analyzed by Western blot; the single cell gel electrophoresis was used to detect the level of deoxyribonucleic acid damage (DNA damage). The results suggest that UEMAM significantly increased survival time, and decreased body weights and abdomen circumference over the saline control group. The treatment increased cell apoptosis, cycle arrest and DNA damage compared to the saline control group. UEMAM significantly enhanced the therapeutic effect of heavy ion beam radiation in survival time, and decreased body weights and abdomen circumference in the tumor-baring mice. The combination increased cell apoptosis, cycle arrest and DNA damage compared to the radiotherapy group. The results of Western blot suggest that the treatment significantly enhanced p53-induced apoptotic signals. The experiment discovered that UEMAM could improve radiosensitivity of H22 ascitic tumor through activation of p53-mediated apoptotic signal pathway.

OBJECTIVE:To optimize the extraction technology of Yigu granule. METHODS:L9(34)orthogonal test was used, using comprehensive scores of transfer rate of icariin,gentiopicroside,loganic acid and yield rate as evaluation indexes,alcohol volume fraction,the amount of solvent,extraction times and extraction time as investigation factors,extraction technology of Yigu granule was optimized,and the verification test was conducted. RESULTS:The optimized extraction technology was 10-fold 60%ethanol reflux extraction for twice,1 h once. Results of verification test showed,average transfer rates of icariin,gentiopicroside and loganic acid were 81.28%(RSD=1.54%,n=3),48.71%(RSD=2.37%,n=3)and 59.82%(RSD=2.52%,n=3);aver-age yield rate was 31.48%(RSD=1.97%,n=3). CONCLUSIONS:The optimized extraction technology for Yigu granule is sta-ble and feasible with good reproducibility,which can provide basis for the follow-up study of production process.

Objective:This study was conducted to evaluate the roles of internal mammary sentinel lymph node biopsy (IM-SL-NB) in the treatment of breast cancer patients with clinically positive axillary lymph nodes. Methods:This study is a one-armed clini-cal research conducted from June 2013 to October 2014. A total of 64 breast cancer patients from Shandong Cancer Hospital with clini-cally positive axillary lymph nodes were enrolled in the study. All patients underwent axillary lymph node dissection. Meanwhile, IM-SLNB was performed in all patients using the new injection method of radiotracer. Results:Among the 64 enrolled patients, the visual-ization rate of internal mammary lymph node was 59.4%(38/64). For the 38 patients who were subjected to visualization of the internal mammary node, the detection rate was 100%(38/38), and the incidence of complications was 7.9%(3/38). The metastasis rate of inter-nal mammary lymph node was 21.1%(8/38). Patients with upper inner quadrant tumors and metastasis of more axillary lymph nodes had a significantly higher chance of developing sentinel lymph node metastasis (P<0.001 and P=0.017, respectively) than the other pa-tients. The clinical benefit rate of the above mentioned treatment was 59.4%. Among the patients, 12.5%(8/64) received extra internal mammary radiotherapy, whereas 46.9%(30/64) patients avoided the unnecessary internal mammary radiotherapy. Conclusion:IM-SL-NB should be performed in breast cancer patients with clinically positive axillary lymph nodes because IM-SLNB could provide the ac-curate indication of radiation to the internal mammary area, especially for the patients with upper inner quadrant tumors and those with a suspiciously high level of axillary lymph node metastasis.

Background and purpose:Whether axillary sentinel lymph node biopsy (ASLNB) could replace axillary lymph node dissection (ALND) in patients who converted after neoadjuvant chemotherapy (NAC) from cN+ to ycN0 is still contentious, and the previous study only evaluated the pathological status of ALN without internal mammary lymph node (IMLN) condition. This study is to evaluate roles of ASLNB and internal mammary sentinel lymph node biopsy (IM-SLNB) in breast cancer patients after NAC.Methods:From Jan. 2012 to Dec. 2014, 60 breast cancer cT1-4N0-3M0patients who were scheduled for neoadjuvant chemotherapy (NAC) and agreed to accept surgery after NAC from our department were enrolled into the retrospective study. Patients with cN0 before NAC and ycN0 after NAC underwent ASLNB (group A). Patients with cN+ received NAC and ycN0 after NAC (group B) were treated with ASLNB and ALND. Only patients whose clinical nodal status remained positive (ycN+) after NAC underwent ALND without ASLNB (group C). All the patients received radiotracer injection and patients in group A and group B received blue dye injection additionally. Meanwhile, IM-SLNB would be performed for all patients with IM-SLN visualization.Results:The number of patients enrolled in group A, group B and group C was 6, 45 and 9 cases respectively. The accuracy rate of ASLNB in group A was 100% (6/6). Only one patient was axillary sentinel lymph node (ASLN) positive performed ALND. With combination of blue dye and radiolabeled colloid, the accuracy rate of ASLNB in group B was 100% (48/48) and the false negative rate (FNR) was 17.9% (5/28). The FNR in patients with 1, 2 and>2 SLNs examined was 27.3% (3/11), 20.0% (2/10) and 0% (0/7). All of the ALNs were positive in group C. The visualization rate of IM-SLN was 63.3% (38/60). The detection rate of IM-SLNB was 97.4% (37/38) and the metastasis rate was 8.1% (3/37). The incidence of complications was 5.3% (2/38).Conclusion:ASLNB can be performed either before or after preoperative chemotherapy for patients with cN0 disease. Among women with cN+ converted to ycN0 who had 3 or more SLNs examined, the FNR could return to be less than 10%. Those patients whose nodes are still ycN+ should perform ALND. IM-SLNB should be performed routinely in all breast cancer patients after NAC, for it might help to make clear of the nodal staging and the pathological status of IM-SLN and provide the accurate indication of radiation to the internal mammary area in case of under-stage and under-/over-treatment, expecting to develop the deifnition of pathological complete response (pCR).

Background and purpose:Sentinel lymph node biopsy has replaced axillary lymph node dissec-tion as the standard staging procedure in early breast cancer patients with clinically negative axillary lymph nodes. It is a critical step for staging and treatment. This study investigated the localization effect of a novel tracer for breast cancer sentinel lymph node biopsy [indocyanine green (ICG)-rituximab (R)], using the hind limb drainage in mice as an animal model.Methods:For exploring the optimal dose and imaging time, different doses of ICG-R were injected subcutane-ously to the dorsum of the foot in the BALB/c mice. Then the lfuorescence vasculature imaging instrument was used continuously to observe the popliteal fossa lymph node (as sentinel lymph node) from the injection to 3 h after injection. For exploring the sustained localization effect, the optimal dose of ICG-R was injected and the imaging instrument was used from imaging to 24 h after injection.Results:The time from injection to imaging and the time from injection to the optimal imaging were shortened with the increased doses, and the imaging rate of the second or third level node increased accordingly. The best dosage of the novel tracer was 0.12 μg (dosage of indocyanine green) and the time from injection to the optimal imaging was about 34 min. After the observation for 24 h, the imaging rate of sentinel lymphnode was maintained at 100%, and the imaging rate of the second and the third level lymph node increased from 0% to 20% and 10%, respectively.Conclusion:ICG-R could clearly locate the sentinel lymph node. There is no imaging of the second level lymph node within 6 h. The novel tracer has high value in the clinical application.

ObjectivesTo identify the related abnormalities of gray matter in pediatric patients with Tourette syndrome (TS) by using the optimized voxel-based morphometry (VBM).Methods Three dimensional T1WI was acquired in 31 TS children (28 boys,3 girts,mean age 8 years,range 4-15 years) and 50 age- and sex-matched controls on a 1.5 Tesla Philips scanner. Images were pre-processed and analyzed using a version of VBM 2 in SPM 2.The whole brain gray matter volume was compared between the study and control group by using t-test.Multivariate linear regression analysis was used for analyzing the correlation between the change of grey matter volume within each brain region (mm3 ) and YGTSS score and course of disease of TS patients.Statistical analyses were performed by using SPSS 13.0.ResultsUsing VBM,significant increases in gray matter volumes in left superior parietal lobule, right cerebellar hemisphere and left parahippocampal gyrus were detected in TS patients,and the volume changes were 4059,2126 and 84 mm3 ( t =3.93,3.71,3.58,P ＜ 0.05 ) respectively.Compared to the control group,decreased grey matter volumes were found in medulla and left pons,and the volume changes were 213 and 117 mm3( t =3.53,3.48,P ＜ 0.05 )respectively.Tic severity was not correlated with any volume changes of gray matter in brain (P ＞ 0.05,a small volume correction,KE ≥ 10 voxel).Tic course was negatively correlated with the gray matter volume of left parahippocampal gyrus ( Beta =- 0.391,P =0.039 ).ConclusionsUsing VBM technique,the gray matter abnormalities can be revealed in TS patients without obvious lesions on conventional MR imaging.The increasing volume of temporal and parietal lobes and cerebellar may be an adaptive anatomical change in response to experiential demand. The gray matter volume of the parahippocampal gyrus may be used as one potential objective index for evaluating the prognosis of TS.

Objective To verify the accuracy of the modified technique for internal mammary sentinel lymph node biopsy ( IM-SLNB) in breast cancer .Methods In the validation study , the radiotracer was injected with the modified technique , and fluorescence tracer was injected into the peritumoral breast tissue .The radioac-tive IM-SLN was identified by preoperative lymphoscintigraphy and γprobe.The radioactive IM-SLN received bi-opsy during operation .The status of the fluorescence tracer was identified by the fluorescence imaging system . Results A total of 162 patients were enrolled from Sep .2013 to Dec.2014.IM-SLNB was performed in 110 pa-tients.The radiotracer and the fluorescence tracer were identified in the same IM-SLN in 94 cases, and the con-cordance rate was 85.5%(Case-base, Spearman coefficient correlation 0.823, P<0.001).Conclusion Dif-ferent tracers injected into the different sites of the intra-parenchyma can reach the same IM-SLN, proving the ac-curacy of the modified technique and the hypothesis of IM-SLN lymphatic drainage pattern ( IM-SLN receives not only the lymphatic drainage from the primary tumor area but the entire breast parenchyma ) .