ABSTRACT
The research on human hepatobiliary development and disorders has been constrained by minimal access to human fetal tissue, and low accuracy of animal models. To overcome this problem, we have established a system for the differentiation of human pluripotent stem cells (hPSCs) into functional hepatobiliary organoids (HBOs). We have previously reported that our 45-d approach closely mimics key stages of hepatobiliary development, starting with the differentiation of hiPSC into endoderm and a small part of mesoderm, and subsequently into hepatoblast-like cells, followed by the parallel generation of hepatocyte-like cells and cholangiocyte-like cells, formation of immature HBO expressing early hepatic and biliary markers, and mature HBO displaying hepatobiliary functionality. In this study, we present an updated version of our previous protocol, which only needs 35 days to achieve maturation in vitro. Furthermore, a hepatobiliary culture medium is developed to functionally maintain the HBOs for more than 1.5 months. The capacity of this approach for producing large amounts of functional HBOs and enabling long-term culture in vitro holds promise for applications on developmental research, disease modeling, as well as screening of therapeutic agents.
ABSTRACT
Objective To investigate the analgesic efficacy of ultrasound?guided adductor canal blockade (ACB)after minor arthroscopic knee surgery. Methods Sixty patients undergone minor arthroscopic knee surgery were randomly divided into group ACB(n=20)and group Control(n=20). All patients received spinal anesthesia. The patients in group ACB received ultrasound?gGuided ACB with 20 ml 0.5% ropivacaine,and patients in group Control received 20 ml saline after the surgery. In addition ,all patients have a basic analgesic regimen with etoricoxib. Visual analogue scales(VAS) during rest and passive movement ,additional analgesic dose and side effects were recorded at 4,8,12,24 h Post?operation. At 24 h post?operation,the numbers of patients who can walk for 5 meters were recorded. Results VAS during rest and movement at 4 h,8 h and 12 h post?operation in group ACB were significantly lower than those in group Control. And all patients could walk 5m at 24 h post?operation. No headache,nausea and vomiting,urinary retention and other adverse reactions were observed in group ACB. There were four patients who received additional analgesic and one patient vomitted. Conclusions Significant analgesic effect of the ACB could be detected after minor arthroscopic knee surgery ,with less reduction in requirements for supplemental opioids.
ABSTRACT
Objective To investigate the effect of Akt pathway inhibitor AZD5363 on cell proliferation and invasion of QBC939 and RBE cholangiocarcinoma cells and the mechanism.Methods Western blotting was used to detect Akt and downstream protein and mTOR protein expression in two cancer cell lines after process by AZD5363.Inhibition rate and cytotoxicity was tested by CCK-8 assay,and Transwell assay was used to evaluate the invasive ability of cancer cells.Results QBC9393 cell exposed to AZD5363 LD50 drug concentration (24 ±9) was significantly different compared with control group (t =4.47,P ＜ 0.05),RBE cells LD50 drug concentration (21 ± 8) was significantly different compared with control (t =4.41,P ＜ 0.05).Tumor invasion capacity of QBC939 in drug concentrations of 20 μmol/L (63 ± 12) and 0 μmoL/L (271 ± 27),the difference was statistically significant.RBE exposed AZD5363 upon drug concentrations of 20 μmol/L (58 ± 23) and 0 μmol/L (235 ± 21),the difference was statistically significant.AZD5363 promotes phosphorylation of mTOR in QBC939.Conclusions AZD5363 inhibits the proliferation and migration,inhibiting the phosphorylation of Akt and its downstream molecules.AZD5363 promotes phosphorylation of mTOR in QBC939.