Study on the anti-sepsis mechanism of ursolic acid by targeting myeloid differentiation protein-2 / 中华危重病急救医学

OBJECTIVE@#To explore the mechanism of ursolic acid in treating sepsis using myeloid differentiation protein-2 (MD-2) as the research carrier.@*METHODS@#The affinity of ursolic acid and MD-2 was determined by biofilm interferometry technique, and the bonding mode between ursolic acid and MD-2 was tested with the aid of molecular docking technique. Raw 264.7 cells were cultured in RPMI 1640 medium and subcultured was conducted when the cell density reached 80%-90%. The second-generation cells were used for in the experiment. The effects of 8, 40 and 100 mg/L ursolic acid on cell viability were assessed by methyl thiazolyl tetrazolium (MTT) method. Cells were divided into blank group, lipopolysaccharide (LPS) group (LPS 100 μg/L) and ursolic acid group (100 μg/L LPS treatment after addition of 8, 40 or 100 mg/L ursolic acid). The effect of ursolic acid on the release of cytokines nitric oxide (NO), tumor necrosis factor-α (TNF-α) and interleukins (IL-6, IL-1β) were evaluated by enzyme-linked immunosorbent assay (ELISA). The influence of ursolic acid on the mRNA expressions of TNF-α, IL-6, IL-1β, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were detected by reverse transcription-polymerase chain reaction (RT-PCR). The implication of ursolic acid on the protein expressions of LPS-Toll-like receptor 4 (TLR4)/MD-2-nuclear factor-κB (NF-κB) pathway were tested by Western blotting.@*RESULTS@#Ursolic acid could bind to the hydrophobic cavity of MD-2 through hydrophobic bond with the amino acid residues of the protein. Therefore, ursolic acid showed high affinity with MD-2 [dissociation constant (KD) = 1.43×10-4]. The cell viability were decreased slightly, with the concentration of ursolic acid increasing, and the cell viability of 8, 40 and 100 mg/L ursolic acid were 96.01%, 94.32% and 92.12%, respectively, and there was no significant difference compared with the blank group (100%). Compared with the blank group, the cytokine level of the LPS group was significantly increased. The level of cytokines were significantly reduced by the treatment of 8, 40 and 100 mg/L ursolic acid, and the higher the concentration, the more obvious effect [compared between 100 mg/L ursolic acid group and LPS group: IL-1β (μmol/L): 38.018±0.675 vs. 111.324±1.262, IL-6 (μmol/L): 35.052±1.664 vs. 115.255±5.392, TNF-α (μmol/L): 39.078±2.741 vs. 119.035±4.269, NO (μmol/L): 40.885±2.372 vs. 123.405±1.291, all P < 0.01]. Compared with the blank group, the mRNA expressions of TNF-α, IL-6, IL-1β, iNOS and COX-2 in the LPS group were significantly increased, and the protein expressions of MD-2, myeloid differentiation factor 88 (MyD88), phosphorylation NF-κB p65 (p-NF-κB p65) and iNOS in the LPS-TLR4/MD-2-NF-κB pathway were significantly up-regulated. Compared with the LPS group, the mRNA expressions of TNF-α, IL-6, IL-1β, iNOS and COX-2 were significantly reduced by the treatment of 100 mg/L ursolic acid bound with MD-2 protein [TNF-α (2-ΔΔCt): 4.659±0.821 vs. 8.652±0.787, IL-6 (2-ΔΔCt): 4.296±0.802 vs. 11.132±1.615, IL-1β (2-ΔΔCt): 4.482±1.224 vs. 11.758±1.324, iNOS (2-ΔΔCt): 1.785±0.529 vs. 4.249±0.811, COX-2 (2-ΔΔCt): 5.591±1.586 vs. 16.953±1.651, all P < 0.01], and the proteins expressions of MD-2, MyD88, p-NF-κB p65 and iNOS in the LPS-TLR4/MD-2-NF-κB pathway were significantly down-regulated (MD-2/β-actin: 0.191±0.038 vs. 0.704±0.049, MyD88/β-actin: 0.470±0.042 vs. 0.875±0.058, p-NF-κB p65/β-actin: 0.178±0.012 vs. 0.571±0.012, iNOS/β-actin: 0.247±0.035 vs. 0.549±0.033, all P < 0.01). However, there was no difference in protein expression of NF-κB p65 among the three groups.@*CONCLUSIONS@#Ursolic acid inhibits the release and expression of cytokines and mediators and regulates LPS-TLR4/MD-2-NF-κB signaling pathway by blocking MD-2 protein, and thus plays an anti-sepsis role.

Characteristics of skin microbiota in an atopic dermatitis-like mouse model induced by 2,4-dinitrochlorobenzene / 中华皮肤科杂志

Objective:To investigate characteristics and changes of skin microbiota in atopic dermatitis-like mouse models induced by different concentrations of 2,4-dinitrochlorobenzene (DNCB) .Methods:Totally, 30 male specific-pathogen-free BALB/c mice were randomly divided into 3 groups by using a random number table: negative control group topically treated with 200 μl of mixture of acetone and olive oil at a volume ratio of 3∶1 on the back twice a week for 6 consecutive weeks; high-and low-concentration DNCB groups both topically treated with 200 μl of 1% DNCB on the first and third day at the first week, followed by topical application of 200 μl of 0.5% and 0.1% DNCB, respectively, twice a week for 5 weeks from the second week. Twenty-four hours after the last treatment, the severity of skin lesions was evaluated, and the transepidermal water loss and stratum corneum hydration were measured. After the experiment, the mice were sacrificed, and skin tissues were resected from the back of the mice for histopathological examination. Full-thickness skin tissue samples were obtained from the back of 3 mice in each group. Illumina Miseq PE300 high-throughput sequencing was performed to sequence the V3-V4 variable region of 16S rRNA gene of skin microbiota on the back of the mice, and the composition and structure of the skin microbiota and changes in the relative abundance of different genera were analyzed. One-way analysis of variance was used to analyze differences in indices among the 3 groups, and the Games-Howell method was used for multiple comparisons.Results:The severity scores of skin lesions were significantly higher in the high-and low-concentration DNCB groups (9.83 ± 2.45 points, 2.71 ± 0.56 points, respectively) than in the negative control group (0.51 ± 0.12 points, t=-7.19,-2.85, respectively, both P < 0.05) . Compared with the negative control group, the high-and low-concentration DNCB groups showed significantly increased transepidermal water loss ( t=-7.72,-2.68, respectively, both P < 0.05) , but significantly decreased stratum corneum hydration ( t=6.77, 5.99, respectively, both P < 0.05) ; the transepidermal water loss was significantly higher in the high-concentration DNCB group than in the low-concentration DNCB group ( t=2.76, P < 0.05) , while no significant difference in the stratum corneum hydration was observed between the high-and low-concentration DNCB groups ( P > 0.05) . There was a significant difference in the relative abundance of Corynebacterium among the 3 groups ( F=249.85, P < 0.001) , which was highest in the high-concentration DNCB group. No significant differences in the observed species and Chao1 index of the skin samples were observed among the 3 groups (both P > 0.05) , and the Shannon index was significantly lower in the high-concentration DNCB group than in the low-concentration DNCB group and negative control group ( t=6.96,-6.37, respectively, both P < 0.05) . Conclusion:DNCB could induce atopic dermatitis-like dermatitis in mice, and the severity of skin lesions and degree of barrier function impairment were related to the concentration of DNCB; the species diversity of skin microbiota markedly decreased in the high-concentration DNCB group, indicating that high-concentration DNCB modeling has more advantages in studying microbiological changes associated with atopic dermatitis.

Anhydrous Ethanol Improves Efficiency of Radiofrequency Ablation for the Treatment of Benign Thyroid Nodules:A Prospective Randomized Controlled Trial / 中国医学科学院学报

To investigate the value of injecting a small amount of absolute ethanol into the benign solid nodules of the thyroid before radiofrequency ablation(RFA)to improve the efficiency of radiofrequency ablation. A total of 98 eligible patients(98 nodules)with pathologically confirmed benign solid nodules who were treated in our center from December 2016 to February 2018 were included and randomized into ethanol ablation(EA)combined with radiofrequency ablation(RFA)group(EA+RFA group)and RFA group,with 49 patients in each group.Routine ultrasound,contrast-enhanced ultrasound(CEUS),and thyroid function test were performed before treatment and 1,3,6,and 12 months after treatment.The general information,treatment time,ablation energy,ablation power,postoperative nodule volume reduction ratio(VRR),symptom score(SS)and cosmetic score(CS),thyroid function level,and incidence of complications were compared between these two groups. The mean treatment time [(441.30±243.31)s (790.70±349.82)s;= 4.403, =0.000],mean ablation energy [(3.92±2.01)kJ (5.15±2.12)kJ;=2.709, =0.009],and mean ablation power [(6.07±1.44)W (7.30±1.29)W;=3.612, =0.006] were significantly lower in the EA+RFA group than in the RFA group.At 3,6 and 12 months after surgery,the VRR in the EA+RFA group was(57.73±11.07)%(=-3.16, <0.001),(64.40±10.56)%(=-5.45, <0.001),and(77.29±8.48)%(=-10.46, <0.001),respectively;the VRR in the RFA group was(55.44±13.01)%(=-1.76, <0.001),(65.28±11.33)%(=-5.09, <0.001),and(75.17±9.84)%(=-8.93, <0.001),which were significantly smaller than those before surgery.There was no significant difference in VRR between the EA+RFA group and the RFA group at 1(=3.41, =0.33),3(=2.05, =0.21),6(=2.77, =0.49),and 12 months(=5.05, =0.10)after treatment.During the follow-up,no recurrence of nodules was observed on CEUS.In the EA+RFA group,the SS [(1.77±0.86).(5.54±2.15);=9.63, <0.001] and the CS[(1.39±0.77).(3.32±0.61);=10.09, =0.004]at 12 months after surgery were significantly lower than those before surgery.In the RFA group,SS [(1.63±1.04).(5.90±1.79);=12.72, <0.001] and CS [(1.64±0.83).(3.15±0.72);=8.13, =0.012] at 12 months after surgery were also significantly lower than those before surgery.The CSS in the EA+RFA group was significantly lower than that in the RFA group [(0.93±0.55).(2.44±0.53);=-11.70, =0.007].Both groups had no significant change in thyroid function during the follow-up period,and no serious complications were observed. Anhydrous alcohol injection can effectively improve the efficiency of radiofrequency ablation in treating benign solid thyroid nodules and is effective in reducing nodule volume,alleviating compressive symptoms,and decreasing cosmetic discomfort.

Comparative study on the diagnostic value of multiparametric transrectal ultrasound and multiparametric MRI in prostate cancer / 中华泌尿外科杂志

Objective@#The aim of the study is to compare the diagnostic value of multiparametric transrectal ultrasound(TRUS) and multiparametric magnetic resonance imaging (MRI) in prostate cancer.@*Methods@#The clinical data of 102 patients who received multiparametric TRUS (including conventional transrectal ultrasound, shear wave sonoelastography and contrast enhanced ultrasound), multiparametric MRI(including T2 weighted diffusion weighted, and dynamic contrast enhanced MRI) and laboratory tests from April 2016 to May 2018 were retrospectively analyzed. The average age was 66.1 years old, ranging 38.0-85.0 years old. The average PSA was 30.1 ng/ml, ranging 0.4-227.0 ng/ml. The average PSAD was 0.67 ng/ml2, ranging 0.02-4.27 ng/ml2. The pathology results from TRUS guided biopsy or surgical operation were chosen as gold standard. Diagnostic performance including sensitivity, specificity, positive predictive value(PPV), negative predictive value(NPV), accuracy and area under the receiver operating characteristic curve(AUROC)of multiparametric TRUS and multiparametric MRI in prostate cancer were analyzed.@*Results@#There were 62 prostate cancer and 40 BPH patients in our study. Parallel multiparametric TRUS diagnosed 63 prostate cancer and 39 BPH, and multiparametric MRI diagnosed 75 prostate cancer and 27 BPH. The sensitivity, specificity and accuracy of parallel multiparametric TRUS were 98.4%, 70.0% and 87.3%, respectively. And those of multiparametric MRI were 95.2%, 60.0% and 81.4%, respectively. The AUROC of parallel multiparametric TRUS and multiparametric MRI were 0.842 and 0.776, with no significant differences(P=0.208).@*Conclusion@#The diagnostic value of multiparametric TRUS was not inferior to multiparametric MRI in prostate cancer.

Comparative study on the diagnostic value of multiparametric transrectal ultrasound and multiparametric MRI in prostate cancer / 中华泌尿外科杂志

Objective The aim of the study is to compare the diagnostic value of multiparametric transrectal ultrasound(TRUS) and multiparametric magnetic resonance imaging (MRI) in prostate cancer.Methods The clinical data of 102 patients who received multiparametric TRUS (including conventional transrectal ultrasound,shear wave sonoelastography and contrast enhanced ultrasound),multiparametric MRI (including T2 weighted diffusion weighted,and dynamic contrast enhanced MRI) and laboratory tests from April 2016 to May 2018 were retrospectively analyzed.The average age was 66.1 years old,ranging 38.0-85.0 years old.The average PSA was 30.1 ng/ml,ranging 0.4-227.0 ng/ml.The average PSAD was 0.67 ng/ml2,ranging 0.02-4.27 ng/ml2.The pathology results from TRUS guided biopsy or surgical operation were chosen as gold standard.Diagnostic performance including sensitivity,specificity,positive predictive value (PPV),negative predictive value (NPV),accuracy and area under the receiver operating characteristic curve (AUROC) of multiparametric TRUS and multiparametric MRI in prostate cancer were analyzed.Results There were 62 prostate cancer and 40 BPH patients in our study.Parallel multiparametric TRUS diagnosed 63 prostate cancer and 39 BPH,and multiparametric MRI diagnosed 75 prostate cancer and 27 BPH.The sensitivity,specificity and accuracy of parallel multiparametric TRUS were 98.4％,70.0％ and 87.3％,respectively.And those of multiparametric MRI were 95.2％,60.0％ and 81.4％,respectively.The AUROC of parallel multiparametric TRUS and multiparametric MRI were 0.842 and 0.776,with no significant differences (P =0.208).Conclusion The diagnostic value of multiparametrie TRUS was not inferior to multiparametrie MRI in prostate cancer.

The effect and influencing factors of percutaneous radiofrequency ablation for the treatment of benign thyroid nodules / 中华医学超声杂志（电子版）

Objective To explore the curative effect and influencing factors of percutaneous radiofrequency ablation(PRFA)for benign thyroid nodules in order to improve the effect of thyroid benign nodule ablation treatment.Methods The study included 482 benign thyroid nodules.The largest diameter of benign thyroid nodules ranges from 0.7 to 9.2 cm.Two hundred and fifty-eight nodules were solid nodules,224 nodules were cystic-solid nodules,and 96 thyroid nodules combined with the background of Hashimoto's thyroiditis.The patients of this study included 356 females and 126 males,the ages of the study population ranged from 14 to 82 years old.In order to observe the local pain,swelling,skin infection,hoarseness and other complications of the thyroid nodules with PRFA,we conducted conventional ultrasound and contrast-enhanced ultrasound immediately and followed up in 1,3,6,12,18 and 24 months after the treatment.Besides,the nodule volume reduction ratio was calculated,and the effects of gender,age,nodule size,cystic proportion,and the background of Hashimoto's thyroiditis on the treatment were analyzed.Results The volume of benign thyroid nodules after the PRFA treatment was significantly reduced after 1,3,6,12,18,24 months,and their nodule volume reduction ratio(VRR)was(51.2±5.7)%(1 months),(69.7±4.3)%(3 months),(84.6±3.7)%(6 months),(89.3±2.9)%(12 months),(93.7±1.6)%(18 months)and(94.9±1.4)%(24 months),respectively.The thyroid nodules were divided into 3 groups according to the cystic portion,significant differences were found in the volume reduction rate at 1,3,6,12,18 and 24 months after PRFA(F=66.858,69.101,19.410,49.559,146.653 and 309.950 respectively,all P＜0.001),the more cystic portion was,the faster the lesion shrinked.The nodules with the cystic portion ＜ 20%were grouped according to the maximum diameter(≤3 cm/＞3 cm),and the background of Hashimoto's disease; and significant differences were found in the volume reduction rate at 1,3,6,12,18 and 24 months after PRFA(the t values for different size groups were 9.710,8.925,9.899,12.734,17.226 and 42.580,respectively,the t values for whether there is a background of Hashimoto's disease were 66.858,69.101,19.410,49.559,146.653,309.950,respectively,all P ＜ 0.001).The VRR of nodules of which the maximum diameter ≤ 3 cm,or without Hashimoto's thyroiditis was larger after radiofrequency ablation.The nodules of which the maximum diameter≤3 cm and with the cystic portion ＜20%were grouped according to different genders(male or female)and different ages(≤40 or＞40 years old),and the mean VRR of the nodules between different gender and age groups are not significantly different(P＞0.05).Conclusions Ultrasound guided PRFA is a safe and effective minimally invasive therapy for benign thyroid nodules.

Contrast-enhanced ultrasound in the evaluation of thickened wall type of gallbladder carcinoma / 中华医学超声杂志（电子版）

Objective To investigate the contrast enhanced ultrasound (CEUS) characteristics of thickened wall type of gallbladder carcinoma and to evaluate their diagnostic value. Methods The CEUS images, clinical information and pathological results of 26 patients with thickened wall type of gallbladder carcinoma and 37 patients having benign gallbladder disease with thickened wall were retrospectively analyzed. CEUS characteristics of thickened wall type of gallbladder carcinoma were investigated and their diagnostic value was evaluated. The age, length of gallbladder, width and thickness of gallbladder wall were analyzed by t test. The CEUS characteristics of gallbladder wall (intensity of gallbladder wall, mucosal morphology and submucosal hypo-enhancement area)were analyzed by χ2 test.The diagnostic test used ROC (receiver operating characteristic) curve to calculate sensitivity, specificity, positive predictive value, negative predictive value and accuracy. Results The gallbladder wall thickness and patient age in malignant group were all larger than those in benign group[(1.63±0.68)cm vs(0.96±0.55)cm,(63.7±10.1)years old vs (53.2±11.8) years old], with statistically significant difference (t=3.70, 4.32, all P<0.001). In the malignant group, CEUS showed irregular mucosal, hyper-enhancement and sub-mucosal hypo-enhancement area, with statistically significant difference(χ2=48.7,42.9,23.8,OR=9.25,6.17,2.47,all P < 0.001).The sensitivity of irregular mucosa, hyper-enhancement and sub-mucosal hypo-enhancement area were all 100.0%, and their accuracy were 93.7%, 90.5% and 76.2% respectively. If lesions with both irregular mucosa and hyper-enhancement were diagnosed as thickened wall type of gallbladder carcinoma, the diagnostic accuracy could be further improved to 98.4%. Conclusion CEUS can facilitate the differential diagnosis of benign and malignant gallbladder diseases with thickened wall in a high diagnostic value.

Correlation between ultrasonographic features of thyroid nodules and BRAFV600Emutation / 中华医学超声杂志（电子版）

Objective To study the correlation between ultrasonographic features of thyroid nodule and BRAFV600Emutation. Methods A total of 179 patients with 194 suspicious throid nodules were included in this study. They underwent ultrasound, biopsy, pathology and BRAFV600Emutation examination between October 2015 and February 2016 at Chinese PLA General Hospital. The size of nodules were (1.1±0.8) cm. The size, echo, boundary, shape aspect ratio, calcification and capsular invasion of nodules were investigated. The correlation between ultrasonographic features of thyroid nodule and BRAFV600Emutation analyzed by chis-square test and Logistic Regression analysis using statistical data as independent variable, BRAFV600Emutation as dependent variable. Results There were significant different in nodule′s ratio, boundary, capsular invasion characteristic between the BRAFV600Epositive group and the BRAFV600Enegative group(χ2=11.174,45.517,11.046,all P < 0.05),and these signs are possibly associated with BRAFV600Emutation by logistic regression model analysis(OR=2.276,95%CI:1.117-4.638, P < 0.05; OR=8.412, 95%CI: 3.836-18.448,P < 0.001; OR=2.582, 95%CI: 1.138-5.860,P < 0.05). Conclusions The ratio, boundary, capsular invasion characteristic of thyroid nodules are possibly associated with BRAFV600Emutation. These signs can be used to predict BRAFV600Emutation and facilitate subsequent treatment for such nodules.

Ardipusilloside-I stimulates gastrointestinal motility and phosphorylation of smooth muscle myosin by myosin light chain kinase

Ardipusilloside-I is a natural triterpenoid saponin, which was isolated from Ardisia pusilla A. DC. The aim of the study was to evaluate the stimulation of ardipusilloside-I on gastrointestinal motility in vitro and in vivo. The experiment of smooth muscle contraction directly monitored the contractions of the isolated jejunal segment (IJS) in different contractile states, and the effects of ardipusilloside-I on myosin were measured in the presence of Ca²⁺-calmodulin using the activities of 20 kDa myosin light chain (MLC₂₀) phosphorylation and myosin Mg²⁺-ATPase. The effects of ardipusilloside-I on gastro emptying and intestinal transit in constipation-predominant rats were observed, and the MLCK expression in jejuna of constipated rats was determined by western blot. The results showed that, ardipusilloside-I increased the contractility of IJS in a dose-dependent manner and reversed the low contractile state (LCS) of IJS induced by low Ca²⁺, adrenaline, and atropine respectively. There were synergistic effects on contractivity of IJS between ardipusilloside-I and ACh, high Ca²⁺, and histamine, respectively. Ardipusilloside-I could stimulate the phosphorylation of MLC₂₀ and Mg²⁺-ATPase activities of Ca²⁺- dependent phosphorylated myosin. Ardipusilloside-I also stimulated the gastric emptying and intestinal transit in normal and constipated rats in vivo, respectively, and increased the MLCK expression in the jejuna of constipation-predominant rats. Briefly, the findings demonstrated that ardipusilloside-I could effectively excite gastrointestinal motility in vitro and in vivo.

The diagnostic value of thyroid rich blood supply lesions by using contrast-enhanced ultrasound / 中华医学超声杂志（电子版）

Objective To investigate the differential diagnostic value for thyroid rich blood supply lesions by contrast-enhanced ultrasound (CEUS).Methods From February 2016 to January 2015,patients who suffered with thyroid nodules underwent conventional ultrasound and CEUS before biopsy.Sixty-two lesions with high-enhancement features were included in the final data.The enhancement patterns within and around lesions of CEUS were analyzed,and the high-enhancement ranges were compared with lesions sizes displayed on conventional ultrasound imaging.When pathology was regarded as the golden standard,39 nodules of 62 were benign,and 23 were malignant.The difference of CEUS characteristics between benign and malignant lesion was compared using x2 test.Results The homogeneity or not had significant difference between benign and malignant lesions with high-enhancement features (x2=9.78,P=0.002).The features of high-enhancement with cystic area and uneven distribution of high-enhancement area had significant difference between benign and malignant lesions (x2=10.77,P=0.001),and the former were frequently seen in benign lesions.Peripheral regular and irregular rings had significant difference between the benign and malignant lesions (x2=24.33,P ＜ 0.001).All lesions showed large high-enhancement area were malignant (x2=6.52,P=0.01).The diagnostic sensitivity,specificity and accuracy of CEUS were 95.83％,78.95％ and 85.48％ for thyroid rich blood supply lesions,which were better than those of conventional ultrasound (87.5％,68.42％,75.81％).Conclusions There are different features in CEUS between the benign and malignant thyroid lesions with rich blood supply.Heterogeneous high-enhancement,peripheral irregular rings and extensive high-enhancement area were malignant features.

Value of Contrast-enhanced Ultrasound and Conventional Ultrasound in the Diagnosis of Papillary Thyroid Carcinoma with Cervical Lymph Node Metastases / 中国医学科学院学报

Objective To compare the ultrasound features of papillary thyroid carcinoma with or without cervical lymph node metastasis (LNM). Methods Patients suspected of thyroid cancer underwent the conventional ultrasound and contrast-enhanced ultrasound (CEUS) examinations. Patients' age,sex,and ultrasound characteristics of lesions were recorded. With the surgical pathology as the golden standard,the ultrasound features were compared between the cervical LNM group and non-LNM group. Results Of 144 patients,51 had cervical LNM and 93 did not. Patients' ages,sex and number of lesions had no significant difference between two groups (all P>0.05). Tumor with LNM had maximum size greater than 0.85 cm,ill-defined margin (P=0.000),irregular shape (P=0.007),internal heterogeneous echogenicity (P=0.007),microcalcification (P=0.020),internal heterogeneous low-enhancement (P=0.002),peripheral non-enhancement ring (P=0.030),and extracapsular extension (P=0.000). Conclusion Conventional ultrasound and CEUS are helpful for predicting the cervical LNM of PTC. CEUS can obtain more accurate diagnostic results for the extracapsular extension,which contributes to the prediction of cervical LNM.

Construction of a lentiviral vector of FoxM1 shRNA and its transfection into human prostate cancer cell lines in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a recombinant lentiviral vector that co-express green fluorescent protein (GFP) and FoxM1 shRNA and establish a prostate cancer cell line with stable FoxM1 down-regulation.</p><p><b>METHODS</b>Three interfering sequences targeting FoxM1 were designed and inserted into the lentiviral vector pHBLV-U6-ZsGreen-Puro. After identification by DNA sequencing, the lentiviral vectors carrying Foxm1 shRNA were packaged in 293 cells. The lentiviral particles were collected to infect human prostate cancer DU-145 cells, and the transfection efficiency was observed under fluorescence microscope; the interference efficiency was assessed using real-time PCR. DU-145 cells with stable FoxM1 down-regulation were screened with puromycin, and the expression level of FoxM1 was detected by Western blotting and the cell growth was observed using MTT assay. The stably transfected cells were examined for cell apoptosis and cell clone formation capacity with flow cytometry and colony formation assay.</p><p><b>RESULTS</b>DNA sequencing demonstrated successful construction of the 3 FoxM1 shRNA lentivirus vectors. Real-time PCR showed a high interference efficiency of FoxM1 shRNA1 vector, which resulted in obvious down-regulation of FoxM1 in DU-145 cells. Western blotting showed that the expression of FoxM1 protein was decreased in FoxM1 shRNA1 lentivirus-transfected cells, which displayed a suppressed cell proliferation, increased apoptosis rate, and attenuated clonogenic ability.</p><p><b>CONCLUSION</b>We have successfully established a prostate cancer cell model with stable FoxM1 down-regulation, which shows lowered proliferative and clonogenic activities with increased cell apoptosis.</p>

Impact of needle size and sonographic feature on accuracy of ultrasound-guided breast biopsy / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the accuracy of ultrasound-guided 16G and 18G core needle biopsy for detecting ultrasound visible breast lesions with different sonographic features.</p><p><b>METHODS</b>A total of 955 sonographically detected breast lesions examined with ultrasound-guided core needle biopsy (US-CNB) and subsequently surgically excised from July 2005 to July 2012 were retrospectively reviewed. Histological findings of US-CNB and the surgical specimens were analyzed for agreements, sensitivities, false negative rates, and underestimate rates according to different sonographic features.</p><p><b>RESULTS</b>The pathological results of the US-CNB showed malignant lesions in 84.1%, high-risk lesions in 8.4%, and benign lesions in 7.5% of the samples. The overall agreement rates were 92.4% for 16G CNB and 92.8% for 18G CNB; their complete sensitivities and false negative rates were both 98.6% and 1.4%, respectively; the high-risk underestimate rates and DCIS underestimate rates were 48.0% and 46.2% for 16G CNB vs 53.3% and 41.2% for 18G CNB, showing no significant difference between the two groups (P>0.01). For both 16G and 18G CNB, the agreements were better for mass lesions than for non-mass lesions (P<0.01). For the mass lesions with a diameter no greater than 10 mm, the agreement rates were lower than the overall data (P<0.01). Calcification in the lesions did not affect the agreement rates (P>0.01).</p><p><b>CONCLUSION</b>Ultrasound-guided 16G and 18G CNB are both accurate methods for evaluating ultrasound visible breast mass lesions with a diameter larger than 10 mm.</p>

Evaluation of bladder outlet obstruction in benign prostatic hyperplasia patients using shear wave sonoelastography: a prospective study / 中华泌尿外科杂志

Objective To evaluate the correlation between the elastic modulus of prostate and the degree of bladder outlet obstruction (BOO) in benign prostatic hyperplasia (BPH) patients in order to build a new non-invasive diagnostic indicator for BOO.Methods From november 2011 to November 2012,a total of 34 BPH patients were enrolled in this prospective study.Clinical information of the patients including age,international prostate symptom score (IPSS) and quality of life (QOL) were collected.The morphological features of prostate including total prostate volume,transitaion zone volume and transition zone index were detected by transrectal ultrasound (US),and prostate elastic modulus was detected using shear wave sonoelastograhy.After US examination,urodynamic study was performed by the urologist and pressure-flow curves were used to determine the diagnosis and the degree of bladder outlet obstruction.The correlation of prostate elastic modulus and other features including clinical information,morphological features and urodynamic features with the degree of bladder outlet obstruction were evaluated.ROC curves were evaluated.The feature with the best diagnostic performance was obtained,the diagnostic values of which were calculated.Results 9 patients had no BOO,with the average prostate elastic modulus of (27.3±5.0) kPa; 12 patients had mild to moderate BOO,with the average prostate elastic modulus of (30.7±2.8) kPa; 13 patients had severe BOO,with the average prostate elastic modulus of (34.7±2.4) kPa.The prostate elastic modulus had higher correlation coefficient with the degree of bladder outlet obstruction than the other features (r =0.754,P=0.000).It also had the highest AUC of 0.853 (95％CI 0.720-0.986).At a cutoffof 31.65 kPa,the sensitivity,specificity and accuracy of the prostate elastic modulus for the diagnosis of BOO were 72％,90％and 81％ respectively.Conclusion The prostate elastic modulus is highly correlated with the degree of bladder outlet obstruction,which might serve as a novel,promising indicator to evaluate BOO in BPH patients.

Quantitative evaluation of portal pressure by contrast enhanced ultrasonography / 中华超声影像学杂志

Objective To investigate the feasibility of non-invasive quantitative evaluation of portal pressure (Pp) by contrast enhanced ultrasonography (CEUS).Methods 18 portal hypertension patients (PTH group) were performed with CEUS within one week before splenectomy and pericardial devascularization,and 20 healthy volunteers as controls were recruited in this study (control group).Hepatic-right kidney sectionwas chosen to calculate the area under curve of portal vein/hepatic artery (Qp/Qa) and the perfusion intensity of portal vein/hepatic artery (Ip/Ia) through time intensity curves (TIC) of liver parenchyma generated from CEUS images.Pp was measured by intra-operative mesenteric vein catheter,and the correlation betweenPp and Qp/Qa,Ip/Ia were analyzed by Pearson correlation test.Results The levels of Qp/Qa and Ip/Ia in the PTH group were 2.28 ± 0.66 and 0.35 ± 0.14 respectively,which were both significantly declined than that in the controlgroup (5.72 ± 3.69 and 1.97 ± 0.17).In the PTH group,the correlation coefficient were-0.747 and-0.617,and the linear regression equations were Y =-83 X + 5.013 andY =-15X + 0.837,which indicated that Qp/Qa and Ip/Ia had significant correlation with Pp.Conclusions CEUS parameters,including Qp/Qa and Ip/Ia,are significantly correlated to Pp in portal hypertension patients,which indicate that CEUS could be a new non-invasive clinical method for evaluating Pp.

Noninvasive evaluation of hepatic fibrosis stages by contrast enhanced ultrasonography parameters / 中华超声影像学杂志

Objective To discuss the feasibility of non-invasive quantitative evaluation of hepatic fibrosis by contrast-enhanced ultrasonography (CEUS) parameters. Methods CEUS of liver and liver biopsy were performed in 86 patients. CEUS parameters including area under curve of portal vein/hepatic artery(Qp/Qa) ,perfusion intensity of portal vein/hepatic artery(Ip/Ia), decreasing rate of liver parenchyma perfusion (β) and perfusion time of portal vein (Tp) were calculated based on CEUS images. Differences of the four parameters among stages of hepatic fibrosis were analyzed by ANOVA and Spearman rank correlation test was applied to analyze correlation between parameters and hepatic fibrosis stages. Areas under receiver operating characteristic (ROC) curves were calculated to evaluate the diagnostic accuracy of parameters. Results Tp and β had increasing tend while Ip/Ia and Qp/Qa had declining tend from S0 to S4.They were significantly correlated with fibrosis stage( P ＜0.05＝. The areas under ROC curves of Ip/Ia were 0.931 (≥S1) ,0.884(≥S2 ) ,0.820(≥S3 ) and 0.846(S4 ) respectively. The areas under ROC curves of Qp/Qa were 0.914(≥S1),0.813(≥S2),0.845(≥S3) and0.869 (S4) respectively. Conclusions CEUS parameters of liver parenchyma Ip/la, Qp/Qa, Tp and β are related to fibrosis stages. Ip/Ia and Qp/Qa provide a reliable,non-invasive method for evaluating fibrosis stages.