Objective:To explore the role and mechanism of long non-coding RNA KCNQ1 overlapping transcript 1 (LncRNA KCNQ1OT1) in the migration, proliferation and invasion of hepatocellular carcinoma (HCC).Methods:The experimental method was conducted. The expression levels of LncRNA KCNQ1OT1 in HCC tissues and normal liver tissues in the StarBase database were collected. The experimental methods including real-time quantitative PCR, cell transfection, scratch assay, CCK8 assay, Transwell assay, Western blot were used to determine the expression, migration, proliferation, invasion of LncRNA KCNQ1OT1 in HCC cells and its relationship with phosphatidylinositol 3-kinase/phosphorylated AKT Protein (PI3K /p-AKT) signaling pathways. Observation indicators: (1) expression of LncRNA KCNQ1OT1 in HCC tissues and normal liver tissues; (2) the migration of HepG2, SMCC-7721 and MHCC-97H HCC cells after LncRNA KCNQ1OT1 gene knockdown; (3) the proliferation and invasion of HepG2, SMCC-7721 and MHCC-97H HCC cells after LncRNA KCNQ1OT1 gene knockdown; (4) effects of LncRNA KCNQ1OT1 gene knockdown on PI3K/p-AKT signaling pathways. Measurement data with normal distribution were expressed as Mean± SD, and comparison between groups was analyzed using the t test. The Kaplan-Meier method was used to calculate survival rates and draw survival curves. Results:(1) Expression of LncRNA KCNQ1OT1 in HCC tissues and normal liver tissues. The expression levels of LncRNA KCNQ1OT1 in 374 HCC tissues and 50 normal liver tissues from StarBase database were 3.320±0.017 and 1.470±0.025, respectively, showing a significant difference ( t=5.24, P<0.05). Results of gene expression profile interactive analysis showed that the 30-month disease-free survival rates of HCC patients with high and low expression levels of LncRNA KCNQ1OT1 were 41% and 55%, respectively, with a significant difference ( χ2=6.209, P<0.05). The relative expression of LncRNA KCNQ1OT1 in HepG2, SMCC-7721and MHCC-97H cells were 1.470±0.042, 3.300±0.032, 4.040±0.031, respectively, versus 1.000±0.022 in normal liver cells (LO2), showing significant differences ( t=17.66, 95.40, 114.20, P<0.05). (2) The migration of HepG2, SMCC-7721 and MHCC-97H HCC cells after LncRNA KCNQ1OT1 gene knockdown. ① Results of cell transfection showed that the relative expression levels of LncRNA KCNQ1OT1 in HepG2, SMCC-7721 and MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 0.350±0.016, 0.310±0.020, 0.380±0.018, respectively, versus 1.000±0.021, 1.000±0.018, 1.000±0.019 in the negative control cells, showing significant differences ( t=23.40, 28.15, 22.32, P<0.05). ② Results of scratch assay showed that the healing rates of HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 85.0%±1.9%, 75.0%±1.8%, 90.0%±1.7%, respectively, versus 100.0%±2.0%, 95.0%±1.8%, 72.0%±1.7% of the negative control cells, showing significant differences ( t=31.35, 47.36, 38.42, P<0.05). ③ Results of Transwell assay showed that the vertical migration rates of HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 195±10, 205±12, 85±8, respectively, versus 520±11, 430±7, 405±20 of the negative control cells, showing significant differences between them ( t=922.30, 458.20, 708.40, P<0.05). (3) The proliferation and invasion of HepG2, SMCC-7721 and MHCC-97H HCC cells after LncRNA KCNQ1OT1 gene knockdown. ① Results of CCK8 assay showed that 72-hour optical densities of HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 1.370±0.018, 1.240±0.016, 1.360±0.020, respectively, versus 0.900±0.023, 1.740±0.032, 1.230±0.025 of the negative control cells, with significant differences ( t=10.79, 12.00, 7.56, P<0.05). ② Results of Transwell assay showed that the invasion numbers of HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 186±12, 155±7, 75±9, respectively, versus 505±1, 245±8, 300±15 of the negative control cells, showing significant differences ( t=955.90, 163.40, 530.90, P<0.05). (4) Effects of LncRNA KCNQ1OT1 gene knockdown on PI3K/p-AKT signaling pathways. Resluts of Western blot showed that the relative repression levels of PI3K in HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 0.447±0.009, 0.430±0.012, 0.354±0.006, respectively, versus 0.820±0.017, 0.850±0.012, 0.531±0.001 of the negative control cells, showing significant differences ( t=18.94, 25.72, 27.46, P<0.05). The relative repression levels of p-AKT in HepG2, SMCC-7721, MHCC-97H cells after LncRNA KCNQ1OT1 gene knockdown were 0.343±0.015, 0.410±0.012, 0.579±0.006, respectively, versus 0.546±0.012, 0.620±0.012, 0.830±0.012 of the negative control cells, showing significant differences ( t=10.78, 12.86, 19.02, P<0.05). Conclusions:LncRNA KCNQ1OT1 plays an important role in the occurrence and development of HCC. LncRNA KCNQ1OT1 gene knockdown can inhibit the PI3K/AKT signaling pathways, so it can significantly inhibit the proliferation, migration and invasion of HCC cells.

Objective To explore the effect of fish oil fat emulsion as perioperative nutritional support on patients with liver cirrhosis and portal hypertension.Methods Randomized controlled clinical trial was performed between September 2011 and September 2017 in patients with liver cirrhosis and portal hypertension who underwent pericardial devascularization and splenectomy.Hypocaloric total parenteral nutritional support (TPN) started from the first day after the operation for 5 consecutive days.Patients were divided into experimental group and control group according to thetype of fat emulsion used.43 patients in experimental group were applied for fish oil fat emulsion injection (10％ Omegaven) + medium long chain structure fat emulsion (20％STG) and 42 patients in control group were applied for medium long chain structure fat emulsion (20％STG).Liver function (total bilirubin and alanine aminotransferase),nutrition index (serum albumin and prealbumin),inflammatory mediators (TNF-α,IL-6 and IL-10) were measured before and after the operation,and the clinical outcomes were observed.Results There was no statistically significant difference in liver function and nutritional indices between the experimental group and the control group (P＞0.05).The inflammatory mediators like TNF-u,IL-6 and IL-10 on the first day after surgery were significantly higher than those before surgery in both groups [experiment group:(225.54±54.78) vs.(61.49±16.47),(74.94±6.36) vs.(39.84±2.77),(77.53±11.4) vs.(46.05±6.13) ng/L;control group:(229.26±62.15) vs.(63.48±13.76),(77.23±7.83)vs.(40.64±3.34),(73.89±7.97)vs.(44.88±5.72) ng/L;P＜ 0.01].With the progress of time,the proinflammatory factors like TNF-α and IL-6 decreased after the operation and the range of decrease was higher in experiment group than in control group [d4-d 1:(-56.88± 31.63) vs.(-35.96±20.02),(-13.52±5.20) vs.(-6.38±2.84) ng/L;d7-d1:(-150.67±42.58) vs.(-132.79±53.35),(-27.04±8.97) vs.(-20.85±6.38) ng/L;P＜ 0.05].The range of increase in anti-inflammatory media IL-10 was higher in experiment group than in the control group (d4-d1:(14.22±13.08) vs.(5.64±3.58) ng/L;d7-d1:(17.78±5.58) vs.(-37.96±11.43) ng/L;P＜0.05).The incidence of grade Ⅲ complications and total complications (4.7％ vs.21.4％,23.3％ vs.45.2％) and hospitalization time [(10.12 ±1.48) vs.(12.33±2.04) d] in the experimental group were significantly lower than those in the control group (P＜0.05).Conclusions In patients with liver cirrhosis and portal hypertension,perioperative nutritional support of fish oil fat emulsion can reduce systemic inflammatory response and operative complications and promote rapid recovery through its two-way regulation of inflammatory mediators.

Objective To study the clinical use of indocyanine green (ICG) fluorescence imaging in laparoscopic liver surgery.Methods The clinical and pathological data of 68 patients who underwent laparoscopic hepatectomy using the ICG fluorescence imaging technique during the study period from September 2016 to October 2018 in Zhongnan Hospital,Wuhan University were retrospectively analyzed.Analysis was carried out on the surgical methods,fluorescence navigation methods,ICG injection time and dose,tumor characteristics,and pathological studies of the resected specimens.Results Of 68 patients,3 patients were converted to open surgery,and the remaining 65 patients completed the ICG fluorescence laparoscopic hepatectomy.Thirty-two of these 65 patients underwent ICG fluorescent guided laparoscopic anatomical resection of lower hepatic segment / hepatic hemilivers (positive staining in 17 patients,negative staining in 15 patients),with 19 patients successfully staining with ICG(19 / 32,59.4％).Postoperative histopathology showed primary hepatic solid tumors (n=31),secondary liver tumors (n=12),hepatic cysts (n=4),hepatic hemangiomas (n =5),hepatolithiasis (n =12) and hepatic focal nodular hyperplasia (n =1).These lesions were combined with hepatitis B liver fibrosis in 29 patients.Conclusions ICG fluorescence imaging positively impacted on laparoscopic liver surgery.Proper preoperative ICG injection was helpful for the identification,localization and intraoperative surgical guidance of tumors,especially for patients with deep-seated and central tumors.As a consequence,oncological and surgical safety of laparoscopic liver surgery was improved.Targeted visualization of liver segments and surgical navigation using intraoperative ICG injections facilitated accurate and precise resection of anatomical liver segments or hemi-hepatectomies.The use of intraoperative ICG fluorescence technology for hepatic hemangioma,hepatic cyst,intrahepatic bile duct stones and other benign liver lesions,helped to improve safety of surgery.

OBJECTIVETo explore the efficacy of partial resection of puborectalis combined with mutilation of internal anal sphincter(IAS) in the treatment of puborectalis syndrome with high anal pressure.

METHODSTwenty-five cases of puborectalis syndrome with high anal resting pressure in the preoperative examination received the operation of partial resection of puborectalis combined with mutilation of IAS in Zhongnan Hospital of Wuhan University between January 2013 and May 2015. The position of puborectalis was confirmed by touching with the exposure under the transfixion device, and a transverse incision was made by electrotome between 3 and 5 o'clock direction of puborectalis, then partial puborectalis was lifted by vessel clamp at 5 o'clock direction, and about 0.5 cm of muscular tissue was resected. Between 8 to 10 o'clock direction of anal tube, about 1 cm length of transverse incision was made by electrotome, then partial IAS was lifted by vessel clamp and cut off. Preoperative and postoperative 3-month anorectal manometry and defecography were carried out. Wexner constipation score and Cleveland Clinic incontinence score were implemented before surgery and 3, 6, 12 months after operation. This study was registered in the Chinese Clinical Trial Registry (registration number: ChiCTR-ORB-16007695).

RESULTSOf the 25 cases, 18 were male and 7 were female, the average age was 55 years old and the average course of disease was 9 years. Compared with pre-operation, the postoperative 3-month anal resting pressure and maximal squeeze pressure were significantly decreased [(53.56±9.05) mmHg vs. (92.44±7.06) mmHg, (142.80±20.35) mmHg vs. (210.88±20.56) mmHg, respectively, both P=0.000]; anorectal angulation at resting state and forced defecation state increased significantly [(102.32±4.96)degree vs. (95.88±4.01)degree, (117.88±5.95)degree vs. (89.52±3.25)degree, respectively, both P=0.000]. Wexner constipation score of postoperative 3-month, 6-month, 12-month (8.28±3.91, 7.40±3.64 and 8.04±4.74) was significantly lower than the preoperative score (16.00±3.69, all P<0.05), while the score was not significantly different among 3 time points after operation (P>0.05). Cleveland Clinic incontinence score was 0 at postoperative 6 and 12 months, and revealed 20 cases were effective among all the surgical patients(80%).

CONCLUSIONPartial resection of puborectalis combined with mutilation of internal anal sphincter can effectively reduce anal pressure and improve symptoms of outlet obstruction, which is an effective method in the treatment of puborectalis syndrome with high anal pressure.

Anal Canal ; physiopathology ; surgery ; Constipation ; surgery ; Defecation ; Defecography ; Digestive System Surgical Procedures ; methods ; Female ; Gastrointestinal Diseases ; surgery ; Humans ; Male ; Manometry ; Middle Aged ; Muscle Hypertonia ; surgery ; Pelvic Floor ; physiopathology ; surgery ; Pressure ; Treatment Outcome

OBJECTIVE To determine the distribution of allergens in patients with allergic rhinitis in Wenzhou area. METHODS Patients with AR symptoms from January 2013 to December 2014 were given skin prick test (SPT). The clinical data about SPT was retrospectively colleceted to analyze SPT results. RESULTS 1. Among 2991 individuals, the total positive rate of SPT was 82.0%, with Dermatophagoides farinae (Der. f) and Dermatophagoides pteronyssinus (Der.p) as the most common allergens; the positive rate of inhalant allergen was obviously higher than that of ingestive ones, with significantly statistical difference(χ2=2006.557,P<0.01). Most of patients were allergic to double allergens; the intensity of Der.f and Der.p mainly presented as (++++), with no significant difference(Z=-0.391, P=0.696). 2. There was significant difference of variation with seasons(χ2=34.254, P<0.01). 3. No significant difference of positive rate were observed in different AR-courses(χ2=16.102, P<0.01). 4. Significant difference of positive rate was found among different age-groups; The positive rate was increased along with growing age, got peak at group of 10-12 years old, and then got down after that. CONCLUSION Dust mite was the main allergen coursed AR in Wenzhou area. Seasons and age were two important factors effecting on positive of SPT and onset of AR. The positive rate of allergens was related to age.

Objective To investigate the application value of laparoscopic hepatectomy (LH) in the treatment of liver neoplasms. Methods Clinical data of 78 patients with liver neoplasms undergoing hepatectomy in Zhongnan Hospital of Wuhan University from June 2011 to June 2014 were retrospectively analyzed. The patients were divided into the LH group (n=31) and open hepatectomy (OH) group (n=47). The informed consents of all patients were obtained and the local ethical committee approval had been received. Intra-and post-operative situation and liver function of two groups were compared. Normally distributed data of two groups were compared with t test, non-normally distributed data were compared with rank-sum test and the comparison of rate was conducted using Chi-square test. Results Intraoperative blood loss, length of wound, retention time of abdominal drainage catheter, postoperative start time of eating and average postoperative length of hospital stay in the LH group were 150(100-200) ml, 5(4-6) cm, 5(3-6) d, 2(2-2) d and (8±3) d, which were significantly less compared with 300(300-600) ml, 20 (18-20) cm, 8(6-10) d, 3(2-3) d and (12±4) d in the OH group (Z=-5.405,-7.760,-4.366,-3.746;t=-3.608;P<0.05). The hospitalization cost in the LH group was (51±7) thousand yuan, which was significantly higher than (45±10) thousand yuan in the OH group (t=3.198, P<0.05). On the postoperative 1, 3 day, the levels of aspartate aminotransferase (AST) and aspartate aminotransferase (ALT) in the LH group were 94(62-114), 47(42-58) and 116(68-136), 46(39-50) U/L, which were significantly lower than 110(93-158), 152(95-220) and 141(97-236), 49(42-120) U/L in the OH group (Z=-2.416,-6.539,-2.764,-2.229;P<0.05). In the LH group, the serum levels of albumin (ALB) on the postoperative 1, 3 day were (31±6), (35±3) g/L, which were signiifcantly higher compared with (25±5), (34±3) g/L (t=4.958, 2.191;P<0.05). Conclusions LH is an efifcacious and safe treatment for liver neoplasms. Compared with OH, LH has obvious advantages of smaller incision and shorter recovery time.

Objective To investigate the molecular expression and clinical significance of Vimentin and E-cadherin during epithelial-mesenchymal transformation of hepatocellular carcinoma (HCC) tissues.Methods Hepatocellular carcinoma and paracancerous tissue were collected from 55 patients,while nonneoplastic nonnal liver tissue were collected from 20 patients without cancerous diseases as control group.Expressions of Vimentin and E-cadherin were examined by immunohistochemistry staining.Clinicopathological features of the relative expression of Vimentin and E-cadherin,and its influence on prognosis were discussed as well.Results Vimentin was positively expressed in 69.1％ cancer tissue,and was significantly higher than paracancerous tissues and normal liver tissues(P ＜ 0.05).E-cadherin was positively expressed in 27.3％ cancer tissue and was significantly lower compared with paracancerous tissues and normal liver tissues(P ＜0.05).The expression of Vimentin and E-cadherin was negatively correlated(r =-0.653,P ＜ 0.05).Vimentin expressions in subgroups of TNM stage Ⅲ-Ⅳ,poor differentiation,multiple tumor nodules,metastasis and recurrence were all significantly higher than the respective subgroups as TNM stage Ⅰ-Ⅱ] (x2 =7.267,P ＜ 0.05),moderate to well differentiation (x2 =4.045,P ＜ 0.05),single tumor node (x2=12.143,P ＜0.05),without metastasis or recurrence (x2 =7.267,P ＜ 0.05).On the contrary,Ecadherin showed higher expression in the subgroup of single tumor node compared with multiple tumor nodules(x2 =6.878,P ＜ 0.05).The follow-up of patients further demonstrated that patients with positive Vimentin expression had a significantly lower disease-free survival compared with the Vimentin negative group (P ＜0.05).Conclusions Vimentin and E-cadherin both participated in epithelial-mesenchymal transition process of hepatocellular carcinoma.High expression of Vimentin and low expression of E-cadherin were closely correlated with occurrence and development of HCC.Vimentin and E-cadherin may serve as impottant biomarkers in clinical diagnosis and prognosis prediction of HCC.

Hepatolithiasis is a kind of mixed stone disease which mainly depends on pigment stone.However,the etiology is not clear and the pathogenesis is very complex.A lot of studies confirmed that helicobacter pylori played an important role in the occurrence and development of disease.This paper mainly expounds the problems concerning the characteristic of helicobacter pylori and its connection with hepatolithiasis, its state in the formation of hepatolithiasis.

Objective To investigate the effects of radiotherapy combined with resveratrol( Res)on proliferation, invasion and apoptosis of hepatoma cell line Bel-7404. Methods Bel-7404 cell line was treated with Res(25 μmol·L-1 ) combined with radiotherapy,then divided into four groups( group 1:the control;group 2:radiation at dose of 2 Gy;group 3:radiation at dose of 4 Gy;group 4:radiation at dose of 6 Gy). Cell proliferation and invasion were detected by MTT assay. Cell apoptosis were observed by fluorescence microscopy. Expressions of MMP-2 and VEGF proteins were determined by Western blotting. Results Compared with the control group,cell proliferation and invasion were significantly inhibited,while cell apoptosis was increased in all radiation groups(P〈0. 05). Conclusion The sensitivity of hepatoma Bel-7404 cells to radiation can be enhanced by resveratrol. Radiation therapy combined with resveratrol can inhibit proliferation and invasion of hepatoma cells and increase the cell apoptosis,which may be related with the down-regulation of MMP-2 and VEGF proteins.

Objective To study the inhibitory effects of hydroxyapatite nanoparticles on liver VX2 tumor in rabbits after intratumoral injection. Methods 40 rabbits with implantation of liver VX2 tumors were randomly divided into 4 groups and intratumorally injected with different preparations.Group A: (control group), 1 ml nomal saline containing 0.2% CMC-Na; Group B: ( 5-Fu group),20 mg/ml 5-Fu 1 ml; Group C: (Nano HAP), 20 mg/ml Nano HAP 1 ml; Group D: (5-Fu+Nano HAP), 20 mg/ml 5-Fu 1 ml and 20 mg/ml Nano HAP 1 ml. Ultrasonography was performed to measure liver tumor volume 7, 14, 21 d after treatment. Survival durations of the animals were recorded. Tumor tissues and liver tissues close to tumor were obtained and examined histologically.Results The average tumor volumes 7, 14 and 21 d after treatment were (4.93 ±0.76)cm3,(15. 67±2.75)cm3 and (52. 36±10. 57)cm3 in group A, (4. 16±0. 33)cm3 , (10. 26± 1.60)cm3 and (18. 89±4.65)cm3 in group B, (1.43±0.13)cm3 , (3.69±0.77)cm3 and (9.51±2.09)cm3 in group C, (2. 80±0.46)cm3 , (3. 77±0. 91)cm3 and (8. 46±0.95)cm3 in group D respectively. The average tumor volumes of groups B, C and D were significantly smaller than that of group A in the same time phases after treatment. The life span of group C was longer than that of other three groups, and there was no statistically significant difference between group B and group D, although the two groups were significantly longer than group A. Blood flow was not detected by color Doppler or power Doppler in group C and group D. Pathological examination showed that there was obvious intratumoral necrosis in group C and D. Tumor in group B exhibited thoroughgoing necrosis. Conclusion Hydroxyapatite nanoparticles intratumoral injection is safe and feasible for treatment of liver tumor. Hydroxyapatite nanoparticles can exert a significant inhibitory effect on liver VX2 tumor growth in rabbits without liver toxicity.