1.Hydrodynamics-based transgene directively into rat regenerating liver in vivo
Cunshuan XU ; Xuekun XING ; Xianguang YANG ; Qiushi ZHU ; Lei DOU ; Shuaishuai LIU ; You LI ; Fuchun ZHANG
Acta Anatomica Sinica 2009;40(4):599-603
Objective To study the conditions and methods of hydrodynamics-based transgene into rat regenerating liver in vivo. Methods The solution with concentration 30mg/L gene-containing plasmid was injected into rat tail veins at a speed of 2ml/s, then partial hepatectomy (PH) was performed at different times before/after injection, finally the rat (g) and regenerating liver (g) were weighed, and the liver coefficient (Lc) was calculated. Out of 15 groups which are Lc±Lc*0%, *5%, *10%, *15%, *20%, *25%, *30%, *35%, the most suitable group was chosen as correction coefficient to calculate the most appropriate volume of plasmid solution which was injected into the regenerating liver at different recovery times, and at the same time, right lobe of liver was gathered to make frozen section, then observe and quantify the positive green fluorescent protein (GFP) rate at 488 nm excitation wavelength. Results Injection of either physiological saline or empty plasmid has no significant difference compared with control (only PH performance). The appropriate time of hydrodynamics-based transgene is more than 12 hours before PH or anytime after PH. The solution volume of hydrodynamics-based transgene into liver regenerating rat after PH is rat weight (g) ×9%×1/3×corresponding correction coefficient (Trc). Both vector and target gene have effect on the time and abundance of gene expression. Conclusion Hydrodynamics-based transgene can effectively be applied to gene transfection in rat regenerating liver.
2.Effects of ridge-modified titanium alloy surface on activity of fibroblasts
Baobao XUE ; Shuaishuai ZHANG ; Xing LEI ; Yue SONG ; Long BI ; Guoxian PEI
Chinese Journal of Orthopaedic Trauma 2021;23(5):433-438
Objective:To characterize the biological activity of fibroblasts on the surface of titanium alloy sheets with different ridge widths by investigating the effects of ridge widths on the adhesion, proliferation and differentiation of fibroblasts.Methods:Five groups of titanium sheets with ridge widths of 50 μm, 80 μm, 100 μm, 150 μm and 200 μm were prepared, with all the groove depths being 10 μm. The titanium sheets with no ridges were taken as a control group. After fibroblasts were incubated on the sheets, states of their adhesion were observed by scanning electron microscopy (SEM) at different time points. CCK-8 cell proliferation test and immunofluorescence staining were used to observe proliferation and shape of the cells. The effects of ridge widths on adhesion of fibroblasts were evaluated by Vinculin immunofluorescence staining, and the effects of ridge widths on expression of α-smooth muscle actin ( α-SMA) by immunofluorescence. Results:SEM showed that the cells adhered to the ridges on the titanium sheets 48 hours after inoculation. In the groups with smaller ridge widths (from 50 μm to 150 μm), the cells were slender in shape and grew along the ridge direction. CCK-8 indicated that different ridge widths had no significant effect on the proliferation of fibroblasts between the 6 groups ( P>0.05). Immunofluorescence staining showed that the cells arranged in an orderly direction along the ridges; the long axis of the cells in the 50 μm group showed the best consistency with the extending direction of the ridge, with significant differences among the 6 groups ( P<0.05). The Vinculin test found that the secretion of cell adhesion protein was concentrated in the ridge and semi-quantitative analysis showed that the 50 μm group had the most Vinculin secretion, with significant differences among the 6 groups ( P<0.05). The α-SMA test showed that the ridge width had a regulatory effect on the myogenic differentiation of fibroblasts, and the 50 μm group had the strongest expression of α-SMA, with significant differences among the 6 groups ( P<0.05). Conclusions:Modification of ridges on the surface of titanium sheets may affect arrangement, adhesion and myogenic differentiation of fibroblasts. The ridges of 50 μm in width may lead to stronger polarized arrangement of fibroblasts, more secretion of adhesion-related protein and more pronounced myogenic differentiation of fibroblasts.
3.A meta-analysis on the short-term outcomes between laparoscopic versus open splenectomy and azygoportal disconnection in the prevention and treatment of portal hypertension hemorrhage
Jiangen HOU ; Buqiang WU ; Xin DONG ; Wenwei HU ; Jun QIN ; Yanqi LI ; Kun ZHANG ; Shuaishuai XING
Chinese Journal of Hepatobiliary Surgery 2018;24(4):263-267
Objective To systematically evaluate the short-term outcomes of laparoscopic splenectomy and azygoportal disconnection (LSD) with open splenectomy and azygoportal disconnection (OSD) in the prevention and treatment of portal hypertension (PHT) hemorrhage.Methods A meta-analysis was conducted to evaluate the short-term outcomes published in randomized controlled studies and high quality casecontrolled studies on patients who underwent LSD or OSD from January 2006 to January 2016.Results 1 359 patients from 18 Chinese and English articles which met the inclusion criteria were included into this study.LSD and OSD were performed in 685 patients (the LSD group) and 674 patients (the OSD group),respectively.Meta-analysis showed that there was no significant difference in operation time between the LSD and OSD groups [MD=17.66,95%CI=(-2.46 ~37.78),P>0.05].When compared with the OSD group,the LSD group had a shorter postoperative hospitalization stay [MD =-3.99,95% CI =(-4.82 ~ -3.16),P < 0.05],earlier postoperatively first passing of flatus [MD =-1.09,95% CI =(-1.41 ~ -0.78),P < 0.05],less intraoperative bleeding [MD =-272.66,95% CI =(-345.11 ~-200.21),P < 0.05],a lower complication rate [OR =0.34,95% CI =(0.25 ~ 0.47),P < 0.05],less postoperative pain [MD =-2.54,95% CI =(-2.79 ~-2.29),P < 0.05],shorter postoperative ambulation time [MD =-3.16,95% CI =(-3.53 ~-2.79),P < 0.05],less amount of peritoneal drainage [MD =-180.28,95% CI =(-293.06 ~-67.49),P <0.05] and earlier recovery from postoperative loss in appetite [MD =-1.42,95% CI =(-1.70 ~-1.13),P < 0.05].Conclusion Compared with the traditional OSD,LSD had the advantages of less invasiveness,quicker recovery and higher quality of life in the perioperative period.LSD is the preferred operation for the prevention and treatment of PHT massive hemorrhage.