Background Gene transfection is an effective therapeutic avenue to target many kinds of eye diseases.Non-viral vectors with high transfection efficiency,long-term expression,low toxicity and high expression levels are pivotal in gene therapy of corneal disease.Objective This study was to evaluate and compare the safety and efficiency between EntransterTM and liposome vectors for transfer of CD25 siRNA in rat cornea.Methods Eighty male SPF SD rats were randomly divided into EntransterTM-CD25 siRNA group,liposome-CD25 siRNA group,simple CD25 siRNA group and normal saline solution (NSS) group with the right eye as experimental eyes.Corneal epithelia of the rats were completely removed after ocular surficial anesthesia,and 50 μl EntransterTM-CD25 siRNA,liposome-CD25 siRNA,CD25 siRNA solution and NSS were topical administered in the eyes respectively.Ocular response and green fluorescence number on the corneas were examined under the slit lamp assisted microscope 12 hours,24 hours,3 days and 7 days after use of the drugs.The rats were sacrificed and the corneas were obtained,and corneal histopathological examination was performed by using hematoxylin eosin stain.The gene transferred efficiency in the corneas was evaluated by fluorescence technology,and the safety of EntransterTM and liposome carriers was assessed using TUNEL stain.The expression and location of CD11b in the corneas were detected by immunofluorescence technology.The use and care of the experimental animals complied with Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Committee.Results The quantity and intensity of fluorescence staining in the corneas were significantly increased in the EntransterTMCD25 siRNA group in comparison with the liposome-CD25 siRNA group,and the corneal fluorescence appeared earlier in the simple CD25 siRNA group,but it disappeared in 24 hours after transfection.Corneal histopathological examination revealed that the corneal edema and inflammatory cell infiltration in corneal epithelium after gene transfection were more dominant in the liposome-CD25 siRNA group than those in the EntransterTM-CD25 siRNA group,simple CD25 siRNA group and NSS group,and no abnormality was seen in the stroma and endothelium.The number of inflammatory cells was more in the liposome-CD25 siRNA group than that in the EntransterTM-CD25 siRNA group,simple CD25 siRNA group and NSS group (all at P =0.00).The number of apoptosis cells was significantly more in the liposome-CD25 siRNA group than that in the EntransterTM-CD25 siRNA group,simple CD25 siRNA group and NSS group in 12 hours and 3 days after transfection (all at P =0.00).Immunofluorescence assay showed the expression of CD11b primarily located in the corneal epithelial and stromal layers.The expression of CD11b was gradually enhanced over time in the liposome-CD25 siRNA group and peaked in 24 hours after transfection.However,the expression was absent in the EntransterTM-CD25 siRNA group,simple CD25 siRNA group and NSS group.Conclusions EntransterTM nanometer material-mediated transfection of CD25 siRNA in corneas of normal SD rats appears to have high transfection efficiency,low toxicity and slight irritating response to corneas,and EntransterTM vector is currently available for the gene therapy of corneal disease.

OBJECTIVE To examine expression and distribution of fibronectin (FN) in hepatic tissues from patients with hepatitis or cirrhosis, and its correlation with serum hyaluronic acid (HA) contents. METHODS The expression and distribution of FN were detected with immunohistochemical and pathological methods in liver tissues of 66 cases, among which there were 8 cases of normal liver tissues, 58 cases of various types of hepatitis. The levels of HA in patients with liver disease were detected with radioimmunological method. RESULTS Positive cellular rate of FN in hepatocytes and endothelial cells of chronic active hepatitis (CAH) was apparently higher than those in acute hepatitis (AH) group and chrouic persistent hepatitis (CPH) group(P

From August 2019 to December 2019, 74 patients with choledocholithiasis underwent laparoscopic lithotomy, including 68 cases with primary suture of common bile duct and 6 cases with T-tube drainage. The operation time, postoperative length of hospital stay, hospitalization cost and complications of the two groups were compared. The operation time and length of postoperative hospital stay in primary suture were shorter than those in T-tube drainage group[ (88.1±29.9) min vs. (144.2±30.7) min; (3.9±1.5) d vs. (7.2±3.8) d, both P<0.05]; the hospitalization cost was lower[ (29 578±1 072) Yuan vs. (37 468±2 844) Yuan, P<0.05]. There was no significant difference in the incidence of postoperative complications between two groups ( P>0.05). The primary suture seems to be superior to T-tube drainage in laparoscopic lithotomy, however, two methods have different indications and should be selected according to the individual conditions of patients.

Objective To investigate the effect of NS-398 combined with docosahexaenoic acid (DHA) on apoptosis of cholangiocarcinoma QBC939 cells and its mechanism.Methods In vitro,cultured cholangiocarcinoma QBC939 cells were exposed to 0,25,50,100,150 and 200 μmol/L NS-398 with 0,15,30,45,60 and 75 μg/ml DHA respectively.The absorbance of the QBC939 cells were measured by CCK8 and its growth inhibition ratio were calculated.Flow cytometry was applied to detect cell apoptosis.The level of β-catenin and c-myc mRNA and protein were measured by real-time PCR,Western blot and enzyme-linked immunoabsordent assay respectively.Results Exposure to NS-398 combined with DHA suppressed the growth of QBC939 cells.When NS-398 was at 100 μmol/L and DHA at 45 pμg/ml,the relative growth inhibition rate of QBC939 cells was 90％ (F =5.85,P ＜ 0.05).NS-398 combined with DHA promoted QBC939 cells apoptosis at the early stage (F =8.16,P ＜ 0.01).Real-time PCR could detect low β-catenin and c-myc expression in QBC939 cells disposed by NS-398 combined with DHA (F =7.61,P ＜0.01),(F =7.92,P ＜0.01).NS-398 combined with DHA decreased β-catenin (F =7.75,P ＜ 0.01),(F=8.17,P＜0.01) and c-myc (F=8.76,P＜0.01),(F=8.12,P＜0.01) protein expression in QBC939 cells.Conclusion NS-398 combined with DHA promoted apoptosis and inhibit proliferation of cholangiocarcinoma cells QBC939 in vitro possibly through downregulated mRNA and protein expression of β-catenin and c-myc.

Objective: To compare the evaluation effects of abdominal enhanced computed tomography (CT) coronal imaging versus three-dimensional (3D) vascular reconstruction for critical blood vessels in right colon cancer. Methods: The retrospective and descriptive study was conducted. The clinicopathological data of 50 patients with right colon cancer who were admitted to Changhai Hospital Affiliated to Naval Medical University from January to September in 2018 were collected. There were 33 males and 17 females, aged from 33 to 86 years, with an average age of 63 years. All the 50 patients underwent abdominal multi-slice CT examination on the same CT equipment. The CT examination data were analyzed by two-dimensional (2D) coronal imaging and 3D vascular reconstruction. Observation indicators: (1) anatomical type of Henle trunk; (2) the length of Henle trunk and surgical trunk; (3) the positional relationship between ileocolic vein (ICV) and ileocolic artery (ICA). Measurement data with normal distribution were represented as Mean±SD, and count data were represented as absolute numbers. Kappa coefficients were used to measure the consistency between anatomical types of Henle trunk on 2D coronal images and on 3D vascular reconstructed images. Pearson coefficients were used to evaluate the correlation between the length of Henle trunk and surgical trunk on 2D coronal images and on 3D vascular reconstructed images. Bland-Altman method was used to assess the consistency between the length of Henle trunk and surgical trunk on 2D coronal images and on 3D vascular reconstructed images. Results: (1) Anatomical type of Henle trunk: on the 2D coronal images, 43 of 50 patients had the Henle trunk and 7 had no Henle trunk. On the 3D vascular reconstructed images, 44 of 50 patients had the Henle trunk and 6 had no Henle trunk. There were 2, 21, 17, 3 patients classified as type 0, Ⅰ, Ⅱ, Ⅲ of Henle trunk on the 2D coronal images of 43 patients. There were 6, 19, 16, 3 patients classified as type 0, Ⅰ, Ⅱ, Ⅲ of Henle trunk on the 3D vascular reconstructed images of 44 patients. Six patients with no Henle trunk, 2 in type 0, 18 in type Ⅰ, 15 in type Ⅱ, and 3 in type Ⅲ had the same anatomical type of Henle trunk on the 2D and 3D images. The consistency between anatomic types of Henle trunk on 2D coronal images and on 3D vascular reconstructed images was high (κ=0.830, 95% confidence interval: 0.705-0.956, P<0.05). (2) The length of Henle trunk and surgical trunk: on the 2D coronal images, 43 of 50 patients had the length of Henle trunk as (10±5)mm, and 42 of 50 patients had the length of surgical trunk as (34±12)mm. On the 3D vascular reconstructed images, 44 of 50 patients had the length of Henle trunk as (9±5)mm, and 43 of 50 patients had the length of surgical truck as (35±12)mm. The correlation between the length of Henle trunk and surgical trunk on 2D coronal images and on 3D vascular reconstructed images was positive (r=0.872, 0.979, P<0.05). Bland-Altman plot showed a high consistency between the length of Henle trunk and surgical trunk on 2D coronal images and on 3D vascular reconstructed images (P<0.05). (3) The positional relationship between ICV and ICA: on the 2D coronal images, 24 of 50 patients had anterior crossing between ICV and ICA, 26 had posterior crossing between ICV and ICA. On the 3D vascular reconstructed images, 24 of 50 patients had anterior crossing between ICV and ICA, 26 had posterior crossing between ICV and ICA. There was a complete consistency in the positional relationship between ICV and ICA on the 2D coronal images and on 3D vascular reconstructed images. Conclusion Abdominal enhanced CT coronal imaging and 3D vascular reconstruction have the similar evaluation effects for position of critical blood vessels in right colon cancer, with a good consistency.