lncRNA SNHG14 regulates the malignant biological behaviors of thyroid cancer SW579 cells by targeting miR-433-3p / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨lncRNA SNHG14对甲状腺癌SW579细胞恶性生物学行为的影响及其分子机制。方法：收集2017年10月至2018年12月青海省人民医院收治的20例甲状腺癌患者的癌组织及癌旁组织标本，用qPCR检测甲状腺癌组织和对应癌旁组织中SNHG14与miR-433-3p的表达；根据转染物的不同，将SW579细胞分为si-NC组（转染si-NC）、si-SNHG14组（转染si-SNHG14）、miR-NC组（转染miR-NC）、miR-433-3p mimic组（转染miR-433-3p mimic）、si-SNHG14+anti-miR-NC组（共转染si-SNHG14与anti-miR-NC）和si-SNHG14+anti-miR-433-3p组（共转染si-SNHG14与anti-miR-433-3p）。MTT法、FCM、Transwell实验分别检测转染后SW579细胞的增殖能力、细胞周期、细胞凋亡率、迁移及侵袭能力的改变；利用双荧光素酶报告基因实验检测SNHG14是否可结合miR-433-3p，qPCR法检测SNHG14与miR-433-3p之间的相互调控关系。结果：SNHG14在甲状腺癌组织中的表达高于癌旁组织（P<0.05），而miR-433-3p的表达水平低于癌旁组织（P<0.05）。抑制SNHG14的表达或过表达miR-433-3p可使SW579细胞增殖能力降低（P<0.05）、迁移与侵袭细胞数减少（均P<0.05）、细胞凋亡率升高（P<0.05）、G1期细胞比例升高（P<0.05）且S期细胞比例降低（P<0.05）。双荧光素酶报告基因实验证明SNHG14可结合miR-433-3p，抑制SNHG14的表达可提高SW579细胞中miR-433-3p水平（均P<0.05）。同时抑制miR-433-3p 和SNHG14的表达可部分逆转后者对SW579细胞的增殖、凋亡、迁移和侵袭的作用（均P<0.05）。结论：甲状腺癌组织中lncRNA SNHG14呈高表达、miR-433-3p呈低表达，lncRNA SNHG14可通过靶向结合miR-433-3p促进甲状腺癌SW579细胞的增殖、迁移、侵袭而抑制细胞凋亡。

Diagnostic accuracy of contrast-enhanced ultrasound for detecting bland thrombus from inferior vena cava tumor thrombus in patients with renal cell carcinoma

ABSTRACT Purpose: To evaluate the role of contrast-enhanced ultrasound (CEUS) in differentiating bland thrombus from tumor thrombus of the inferior vena cava (IVC) in patients with renal cell carcinoma (RCC). Materials and Methods: We retrospectively investigated 30 consecutive patients who underwent robot-assisted radical nephrectomy with IVC thrombectomy and had pathologically confirmed RCC. All patients underwent US and CEUS examination. Two off-line readers observed and recorded thrombus imaging information and enhancement patterns. Sensitivity, specificity, accuracy, positive predictive value and negative predictive value for bland thrombus were assessed. Results: Of the 30 patients, no adverse events occurred during administration of the contrast agent. Early enhancement of the mass within the IVC lumen on CEUS was an indicator of tumor thrombus. Bland thrombus showed no intraluminal flow on CEUS. There were eight (26.7%) patients with bland thrombus, including three level II, two level III, and three level IV. There were three cases with cephalic bland thrombus and five cases with caudal bland thrombus. Three caudal bland thrombi extended to the iliac vein and underwent surgical IVC interruption. Based on no intraluminal flow, for bland thrombus, CEUS had 87.5% sensitivity, 100% specificity, 96.7% accuracy, 100% positive predictive value and 95.6% negative predictive value. Conclusion: Our study demonstrates the potential of CEUS in the differentiation of bland and tumor thrombus of the IVC in patients with RCC. Since CEUS is an effective, inexpensive, and non-invasive method, it could be a reliable tool in the evaluation of IVC thrombus in patients with RCC.

Evaluation of hepatoprotective activity of Syringa oblata leaves ethanol extract with the indicator of glutathione S-transferase A1

Abstract The leaves of Syringa oblata Lindl., Oleaceae, had been extensively used as a folk medicine to treat various infections, heal inflammations, icteric hepatitis and acute mastitis. The study was designed to evaluate the hepatoprotective activity of S. oblata leaves ethanol extract against CCl4-induced hepatotoxicity in primary hepatocytes and mice with the indicator of glutathione S-transferase alpha 1. The hepatoprotective effects of S. oblata leaves ethanol extract were evaluated on the basis of liver histopathology and biochemical parameters as well as hepatic oxidative stress markers. The results showed that CCl4 negatively modulated biochemical parameters and liver antioxidant activities. However, the use of S. oblata leaves ethanol extract restored altered-serum biochemical parameters and liver antioxidant activities in a dose-dependent manner. Importantly, the trends in S-transferase alpha 1 were similar to alanine aminotransferase and aspartate aminotransferase level, and S-transferase alpha 1 was suggested to be a marker for the evaluation of hepatoprotective activity of S. oblata leaves ethanol extract. Histopathological examination showed that CCl4 causes significant hepatic injury relative to control group. The above findings suggested that S. oblata leaves ethanol extract has hepatoprotective effects against CCl4-induced hepatic injury and S-transferase alpha 1 may be an indicator to evaluate the protective effects of S. oblata leaves ethanol extract.

Essential Oil from Siegesbeckia pubescens Induces Apoptosis through the Mitochondrial Pathway in Human HepG2 Cells / 华中科技大学学报（医学）（英德文版）

Siegesbeckia pubescens (SP) has been used as a traditional medicine for the treatment of and inflammatory diseases.However,the activities of SP against hepatocellular carcinoma and the related mechanisms remain unclear.The present study aimed to examine the effects of the essential oil of SP (SPEO) on the proliferation of hepatocellular carcinoma cells and the possible mechanisms.The growth inhibition of HepG2 cells was analyzed by MTT assay.Hoechst 33258 and fluorescence microscopy were utilized to observe the nuclear morphological changes of apoptotic cells.Flow cytometry was used to detect cell apoptosis and cell cycle.The expressions of the target proteins were detected by Western blotting.The results showed that SPEO obviously inhibited the proliferation of HepG2 cells in a dose-dependent manner.SPEO activated a series of apoptotic proteins in HepG2 cells,increasing expression levels of Bax,caspase-3 and caspase-9,and decreasing the bcl-2 expression level.SPEO displayed promising anti-hepatocellular carcinoma activities in vitro,partly by inducing apoptosis in HepG2 cells through activating the mitochondrial pathway.

Retroperitoneal Laparoscopic Live Donor Nephrectomy: Report of 105 Cases / 华中科技大学学报（医学）（英德文版）

Retroperitoneal laparoscopic live donor nephrectomy offers an intrinsic advantage over conventional transperitoneal laparoscopic nephrectomy because of the potentially lower risk for early and late donor intraperitoneal complications.Herein we presented our experience performing retroperitoneal laparoscopic live donor nephrectomy in 105 donors.All donor nephrectomy was successful.There were no donor deaths and no conversion to open surgery.Mean operation time was 112 min (range,70—200 min).Intraoperative blood loss was 10—150 mL with an average of 30 mL.Warm ischemia time was 1.3 to 6 min with an average of 3.1 min.Postoperative retroperitoneal hematoma occurred in only one case and there were no other surgical complications.Donors were discharged from the hospital 5 to 10 days postoperation.Average postoperative hospital stay was 6.4 days.One graft was removed due to acute rejection.Delayed graft function occurred in two recipients but renal function returned to normal within four weeks.The other recipients had normal renal function in two weeks except three recipients in four weeks.We believe that retroperitoneal laparoscopic live donor nephrectomy is safe,reliable,and less invasive.

Association of Polymorphisms in Angiotensin Ⅱ Receptor Genes with Aldosterone-producing Adenoma / 华中科技大学学报（医学）（英德文版）

This study examined the association of polymorphisms in angiotensin Ⅱ receptor genes (AT1R and AT2R) with the risk for aldosterone-producing adenoma (APA) in a Chinese Han population.Four polymorphisms including rs5182 (573T/C) in exon 4,rs5186 (1166A/C) in 3'-untranslated region (3'-UTR) in AT1R gene and rs5194 (2274G/A) in 3'-UTR,rs1403543 (1675G/A) in intron 1 in AT2R gene were detected in 148 APA patients and 192 normal subjects (serving as control) by using a MGB-Taqman probe.The distribution of genotypes of each locus was in accordance with Hardy-Weinberg Equilibrium (HWE) in the APA and control groups (P＞0.05).The allele A frequency at rs5194 was significantly higher in the APA group (0.49) than in the control group (0.35) (X2=12.08,P=0.001).Subjects with homozygotic genotype AA and heterozygotic genotype GA were at an increased risk for APA as compared to those with GG genotype (OR=2.66,95％ CI=1.45-4.87; OR=1.67,95％ CI=1.02-2.74).Furthermore,rs5194 single-nucleotide polymorphism (SNP) at AT2R gene was significantly associated with APA in additive (OR=1.64,95％ CI=1.21-2.20,P=0.001),dominant (OR=1.94,95％ CI=1.23-3.06,P=0.003),and recessive model (OR=2.01,95％ CI=1.17-3.45,P=0.01).It was concluded that rs5194 polymorphism at AT2R gene was associated with the risk for APA,which may constitute a genetic marker of APA.

Aortic Cell Apoptosis in Rat Primary Aldosteronism Model / 华中科技大学学报（医学）（英德文版）

This study aimed to determine whether aldosterone could induce vascular cell apoptosis in vivo.Thirty-two male rats were randomly divided into 4 groups: vehicle(control),aldosterone,aldosterone plus eplerenone or hydralazine.They were then implanted with an osmotic mini-pump that infused either aldosterone or the vehicle.Systolic blood pressure(SBP)was measured weekly by the tail-cuff method.After 8 weeks,plasma aldosterone concentration(PAC)and renin activity(PRA)were determined by radioimmunoassay.Aortic apoptosis was examined by TUNEL assay.The levels of cytochrome c and caspase-3 were determined by Western blotting and the expression of Bax and Bcl-2 was detected by immnuohistochemistry and Western blotting.The results showed that as compared with control group,aldosterone-infused rats exhibited:(1)an increase in SBP;(2)significantly elevated PAC with depressed PRA;(3)elevated aortic vascular cell apoptosis accompanied with higher levels ofcytochrome c and activated caspase-3; and(4)significantly up-regulated Bax protein with down-regulated Bcl-2.These effects of aldosterone were significantly inhibited after co-administration with eplerenone but not with hydralazine.It was concluded that aldosterone inducedvascular cell apoptosis by its direct effect on the aorta via mineralocorticoid receptors and independently of blood pressure,which may contribute to aldosterone-mediated vascular injury.

Expression of Angiotensin Ⅱ Receptors in Aldosterone-producing Adenoma of the Adrenal Gland and Their Clinical Significance / 华中科技大学学报（医学）（英德文版）

The expression of angiotensin Ⅱ type 1 receptor(AT1R)and angiotensin Ⅱ type 2 receptor (AT2R)in aldosterone-producing adenoma(APA)of the adrenal gland was detected,and their relationship with clinical indexes of APA was analyzed.The mRNA expression of AT1R and AT2R in50 cases of APA and tissues adjacent to tumors and 12 cases of normal adrenal tissues was detected by using reverse transcriptase polymerase chain reaction(RT-PCR).The expression of AT1R and AT2R proteins in paraffin-embedded slices of tissue was detected by immunohistochemistry.The expression of AT1R in adenoma,tissues adjacent to tumor,and normal tissues of the adrenal gland showed no significant differences.The expression of AT2R in APA tissue was lower than that in normal adrenal gland tissues(P<0.05).Correlation analysis of the mRNA expression level of AT2R and clinical data from patients demonstrated that AT2R expression was negatively related to plasma aldosterone concentration(PAC)(r=-0.467,P<0.05),but positively related with plasma renin activity(PRA)(r=0.604,P<0.05).It is concluded that down-regulation of the AT2R expression is possibly related with the tumorigenesis of APA.

The Single Needle Method for Urethrovesical Anastomosis with Strengthened Posterior Fixation during Laparoscopic Radical Prostatectomy / 华中科技大学学报（医学）（英德文版）

The single needle method for urethrovesical anastomosis with strengthened posterior fixation during laparoscopic radical prostatectomy was explored. The method was initiated by performing a fixing suture with a knot at 4 o'clock of the posterior lip of bladder neck,and another suture at nearby position was performed to leave the knot outside. From 5 o'clock to 8 o'clock,sutures were performed every one o'clock to secure posterior approximation,then every two o'clock a suture.To avoid a loose anastomosis,lock sutures were performed every 3 sutures. The needle was always driven full-thickness outside-in in the bladder neck and inside-out on the urethra. After completing the full circumference,the needle was drawn near the 4 o'clock and tied at the tail end. Any leakage could be closed with additional interrupted sutures. The clinical data of 89 patients who underwent this method were retrospectively compared with those of 23 patients who underwent the single knot method. The results showed that the anastomosis,operative and catheterization time was 17.6±4.7min,134.0±10.7 min and 6.5±1.6 days respectively. There were 3 temporal urinary leakages identified in 89 cases requiring prolonged catheterization. No urinary leak and anastomotic stricture was confirmed,and 95.2% patients had total urinary control. It was concluded that this method was simple and safe for urethrovesical anastomosis.

Relationship between the Expression of RASSF1A Protein and Promoter Hypermethylation of RASSF1A Gene in Bladder Tumor / 华中科技大学学报（医学）（英德文版）

To investigate the relationship between the expression of RASSFIA protein and promoter hypermethylation of RASSFIA gene, RASSFIA protein expression was measured by Western blot- ting in 10 specimens of normal bladder tissues and 23 specimens of bladder transitional cell carci- noma (BTCC). The promoter methylation in BTCC and normal bladder tissues was detected by me- thylation-specific PCR (MSP). The results showed that the expression level of RASSFIA protein was significantly lower in BTCC tissues than that in normal bladder tissues. However, it was not corre- lated with its clinical stages and pathological grades. The frequency of promoter methylation of RASSF1A gene was higher in BTCC tissues than that in normal bladder tissues. In 14 patients with the aberrant promoter methylation, 13 showed loss or low expression of RASSF1A protein. It is con- cluded that RASSFIA gene promoter methylation may contribute to the low level or loss of RASSFIA protein expression, the inactivation of RASSFIA gene and the genesis of BTCC. But, it may bear no correlation with its clinical stages and pathological grades.