Schisandrin B Improves the Hypothermic Preservation of Celsior Solution in Human Umbilical Cord Mesenchymal Stem Cells

BACKGROUND@#Human umbilical cord mesenchymal stem cells (hUCMSCs) have emerged as promising therapy for immune and inflammatory diseases. However, how to maintain the activity and unique properties during cold storage and transportation is one of the key factors affecting the therapeutic efficiency of hUCMSCs. Schisandrin B (SchB) has many functions in cell protection as a natural medicine. In this study, we investigated the protective effects of SchB on the hypothermic preservation of hUCMSCs. @*METHODS@#hUCMSCs were isolated from Wharton’s jelly. Subsequently, hUCMSCs were exposed to cold storage (4 °C) and 24-h re-warming. After that, cells viability, surface markers, immunomodulatory effects, reactive oxygen species (ROS), mitochondrial integrity, apoptosis-related and antioxidant proteins expression level were evaluated. @*RESULTS@#SchB significantly alleviated the cells injury and maintained unique properties such as differentiation potential, level of surface markers and immunomodulatory effects of hUCMSCs. The protective effects of SchB on hUCMSCs after hypothermic storage seemed associated with its inhibition of apoptosis and the anti-oxidative stress effect mediated by nuclear factor erythroid 2–related factor 2 signaling. @*CONCLUSION@#These results demonstrate SchB could be used as an agent for hypothermic preservation of hUCMSCs.

Practice of laparoscopic hepatectomy in primary care facilities in China: surgical planning, surgical techniques and postoperative management / 南方医科大学学报

With the development and application of laparoscopic hepatectomy in major medical centers, domestic and foreign guidelines have summarized the indications, surgical techniques and operational procedures of the surgery. But in primary care facilities, where the surgical equipment are available, laparoscopic hepatectomy is performed only in a small number of cases and the progress of its application remains slow. The reasons possibly lie in the failure of a full understanding of the surgery, the lack of anatomical knowledge of laparoscopic hepatectomy, the lack of close multidisciplinary cooperation in the perioperative period and insufficient training of laparoscopic technology. In this review, we elaborate on three aspects of laparoscopic hepatectomy: preoperative planning, surgical techniques and postoperative management. Before the operation, the surgeons should fully understand the anatomical structure of the liver and select appropriate cases considering both the difficulty of operation and the surgical experience of the surgeons. During the operation, the position of the patient and the layout of the stamping card should be appropriate, and the central venous pressure needs to be well controlled in close cooperation with the anesthesiologist. The surgeons should be proficient at the techniques of liver suspension and pulling and at the use of ultrasonic knife, and select correct techniques for management of bleeding and the control of blood flow in and out of the liver. The patient should receive postoperative management with standard enhanced recovery after surgery (ERAS) protocols. These experiences may help to improve the practice of laparoscopic hepatectomy in local hospitals or primary care facilities.

Preparation of a decellularized rat liver scaffold and its biocompatibility / 南方医科大学学报

<p><b>OBJECTIVE</b>To develop a novel method for preparing decellularized liver biological scaffold (DLBS) for liver tissue engineering.</p><p><b>METHODS</b>DLBS was prepared by treatment of rat livers with detergent and enzymatic cell extraction and observed under optical and scanning electron microscopes. To assess the biocompatibility of the product, C3A cells and bone marrow-derived mesenchymal cells (BM-MSCs) were cocultured with DLBS as the scaffold, and the effect of DLBS on the proliferation of C3A cells was evaluated by MTT assay. DLBS was also implanted under the dorsal skin of SD rats to evaluate the tissue biocompatibility of this material.</p><p><b>RESULTS</b>Application of the detergent and enzymatic extraction allowed full extraction of the cells in the liver, leaving an extracellular matrix scaffold composed mainly of collagen and elastic fibrin. The coculture experiment showed that C3A cells and BM-MSCs could grow on and adhere to DLBS. The result of MTT assay showed that DLBS could promote the proliferation of C3A cells.</p><p><b>CONCLUSION</b>This cell-free DLBS, which retains intact extracellular matrix and promotes cell attachment, proliferation, growth and differentiation, can be an ideal biological matrix scaffold material.</p>

Single-port transumbilical laparoscopic cholecystectomy in an adult patient with congenital pulmonary hypoplasia: a case report / 南方医科大学学报

Congenital pulmonary hypoplasia is a rare developmental abnormality of the lung with an incidence rate of around 1/5000 000. As a fatal condition associated with respiratory insufficiency after birth, this disease is rare in adults. We treated a 29-year-old female patient with congenital pulmonary hypoplasia and concurrent of cystic duct calculus in May, 2011 using single-port transumbilical laparoscopic cholecystectomy, which resulted in a good therapeutic effect comparable to that by routine laparoscopic cholecystectomy.

Percutaneous transhepatic biliary radiofrequency and endoprothesis: a new therapy for malignant biliary obstruction / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore a new approach to the management of malignant biliary obstruction using percutaneous transhepatic biliary radiofrequency and endoprothesis.</p><p><b>METHODS</b>Percutaneous transhepatic biliary radiofrequency and endoprothesis were performed in 2 cases of malignant biliary obstruction, including 1 of hilar cholangiocarcinoma and 1 of pancreatic head carcinoma. The tumor was ablated with radiofrequency followed by placement of matched metal stents into the biliary duct.</p><p><b>RESULTS</b>The surgical procedures were carried out smoothly in the 2 cases. The symptoms of the patients were obviously improved after the operation with a significant decrease in the serum levels of total bilirubin, and CA-199 level decreased to the normal level in 1 case.</p><p><b>CONCLUSIONS</b>This new approach is safe for management of malignant biliary obstruction. Compared with the more conventional interventional therapy, radiofrequency can reduce the intraoperative bleeding and arrest the local tumor growth to promote the patency of the stent as well as the postoperative survival of the patients.</p>

Microgravity culture of hepatocytes on cellulose/gelatin macroporous microcarrier / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare macroporous microcarrier Cytopore 2 and Cultispher S for their effects in microgravity culture of CL-1 cells.</p><p><b>METHODS</b>CL-1 cells were cultured on Cytopore 2 and Cultispher S respectively in a rotational cell culture system (RCCS) in a volume of 50 ml. Dynamic morphological observation and cell counting and functional test were carried out during the cell culture.</p><p><b>RESULTS</b>The cells were capable of adhering to and proliferating on both of the microcarriers, reaching the growth peak on day 9 of cell culture with the maximum cell density of 4x10(6)/ml on cultispher S. Albumin was significantly higher in the supernatant of Cytopore 2 than in that of Cultispher S on days 10, 11, and 12 (P<0.05), and the urea level in the supernatant of cytopore 2 was also significantly higher on days 10 and 11 (P<0.05).</p><p><b>CONCLUSION</b>The cells cultured on Cytopore 2, though with a smaller cell number, display better functions than those cultured on Cultispher S under RCCS conditions.</p>

Infusion of autologous mesenchymal stem cells prolongs the survival of dogs receiving living donor liver transplantation / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the mechanism of autologous mesenchymal stem cells (MSCs) in prolonging the survival of dogs receiving living donor liver transplantation.</p><p><b>METHODS</b>Canine models of allogenic living donor liver transplantation was established in 14 beagle dogs by non-venous by-pass method, and in 7 of the recipients, autologous MSCs labeled by BrdU was infused into the portal vein, with the other 7 dogs as the control. The survival time of the two groups of the dogs was observed after the operation. The liver function (AST and ALT levels), liver pathologies and the differentiation of the transplanted cells were also evaluated postoperatively.</p><p><b>RESULTS</b>Compared with the control group, the dogs receiving MSC transplantation showed significantly increased median survival time (P<0.001) with lowered levels of AST and ALT (P<0.01). The two groups exhibited similar graft rejection after the operation. In dogs with MSC transplantation, the BrdU-labeled MSCs differentiated into liver-like cells in the liver and secreted albumin.</p><p><b>CONCLUSION</b>Autologous MSCs infusion through the portal vein during allogenic living donor liver transplantation can prolong the survival of the recipient dogs. The stem cells transplanted can differentiate into mature liver-like cells and secrete albumin in the hepatic tissue.</p>

Induced differentiation of bone marrow stem cells in transplanted rat liver / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the differentiation of bone marrow stem cells in transplanted livers and its impact on the long-term survival of rats with orthotopic liver transplantation.</p><p><b>METHODS</b>Twenty-four female recipient rats with orthotopic liver transplantation were randomized into blank-control group, D-hanks solution group, bone marrow stem cells group with postoperative infusion of stem cells, and the pathological changes of the liver grafts and survival time of the rats were observed. The differentiation of the bone marrow stem cells were assessed 60 days after transplantation using in situ hybridization histochemistry for Sry gene and alpha-fetoprotein (AFP) immunohistochemistry.</p><p><b>RESULTS</b>In rats with postoperative infusion of bone marrow stem cells through the portal vein, the median long-term graft survival time exceeded 180 days, significantly longer than that in the other two groups (P<0.05), and no obvious evidence of acute rejection was observed with positive Sry expression and AFP expression.</p><p><b>CONCLUSION</b>Infusion of bone marrow stem cells through the portal vein following liver transplantation may alleviate acute graft rejection and promote long-term liver graft survival and AFP expression.</p>

PDX-1 gene transfection and expression in bone marrow stromal cells-derived nestin-positive cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To introduce the eukaryotic expression vector pEGFP-C1-PDX-1 into nestin-positive cell derived from bone marrow stromal cells by nucleofection and optimize the conditions for transfection.</p><p><b>METHODS</b>The recombinant plasmid was transfected into bone marrow stromal cells-derived nestin-positive cells with varied DNA quantities or the serum concentration in the medium. The expression of PDX-1 gene in the transfected cells was detected by RT-PCR.</p><p><b>RESULTS</b>Satisfactory efficiency of transfection was achieved with the DNA quantity of 2-10 microg and medium serum concentration of 20%. PDX-1 expression was detected in the transfected cells by RT-PCR.</p><p><b>CONCLUSION</b>The optimized transfection conditions result in enhanced efficiency of PDX-1 gene transfection into nestin-positive cells derived from bone marrow stromal cells, which may serve as the seed cells in tissue-engineering.</p>

Protective effect of donator liver ischemic preconditioning on posttransplant liver graft function in Chinese miniature pigs / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the protective effect of the ischemia preconditioning of the donor liver on posttransplant liver graft function in Chinese miniature pigs.</p><p><b>METHODS</b>Twenty-five partially inbred Chinese miniature pigs were randomized into three groups, namely the normal control group, the ischemia-reperfusion group and ischemic preconditioning group. Biopsies of the liver graft were performed to analyze HSP70 expression by means of immunoblotting, and the changes of serum AST/ALT levels were assayed using an automated biochemical analyzer. Histopathological assessment was carried out to identify the hepatocyte injury using optical and transmission electron microscopy.</p><p><b>RESULTS</b>Ischemia preconditioning resulted in a notable increase in HSP70 expression and milder injury of the hepatocyte microstructure, whereas ischemia-reperfusion caused a significant increase of serum transaminases level (P<0.01) with declined HSP70 expression and obvious microstructural changes of the liver tissue.</p><p><b>CONCLUSION</b>Ischemic preconditioning can produce obvious protective effects on the donor liver, and positively regulates the expression of shock protein.</p>

Protective effect of protein kinase C and mitogen-activated protein kinases and its mechanism in liver ischemic preconditioning / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the protective effects of protein kinase C (PKC) and mitogen-activated protein kinases (MAPKs) their and mechanisms in liver ischemic preconditioning.</p><p><b>METHODS</b>In rat models of liver ischemia-reperfusion (IR) and ischemic preconditioning (IP), the liver function was evaluated by examining serum alanine aminotransferase and aspartate aminotransferase levels, and the morphological changes of the liver cells were observed under microscope. PKC activator phorbol 12-myristate 13-acetate(PMA) and inhibitor chelerythrine(CHE), as well as MEK inhibitor PD98059, were utilized to analyze the phosphorylation of PKC and P44/42 MAPKs.</p><p><b>RESULTS</b>Compared with the control rats, the liver function was best protected in rats of IP group, but not in those of IP group with PD98059 or CHE treatment. The rats in IR group showed improved liver function after PMA treatment. Similarly, the phosphorylation of PKC and P44/42 MAPKs was correlated with the liver function, and highly enhanced PKC and P44/42 MAPKs activity was observed in IP and IR+PMA groups, but decreased activity in IR and IP+CHE groups.</p><p><b>CONCLUSION</b>Phosphorylation of PKC and MAPKs plays a pivotal role in the preservation of the hepatocytes during IP.</p>

Protecting effects of p44/42 MAPK signal transduction pathway on hepatocytes in ischemic preconditioning / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the significance of PKC and p44/42 mitogen-activated protein kinase (MAPK) signal transduction in ischemic preconditioning (IP).</p><p><b>METHODS</b>Through liver cell IP models, PKC inhibitor and MEK inhibitor were utilized to analyze the phosphorylation level of p44/42 MAPK and cell viability was also observed. Rat liver IP models were established which were treated with various drugs. Then the phosphorylation level of p44/42 MAPK in vivo and serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) concentrations were detected. And cellular structures were observed under light microscopy.</p><p><b>RESULTS</b>Similar results were obtained in vivo and in vitro IP models. Compared with the ischemia reperfusion (IR) group in vivo, the phosphorylation level of p44/42 MAPK was obviously increased in IP treated rats (q = 27.217, P < 0.01), and the cellular structure injured slightly. The concentrations of serum ALT and AST in IP group were significantly lower than those in IR group (281.0 U/L +/-35.6 U/L vs 762.8 U/L +/-130.5 U/L and 407.7 U/L +/-73.7 U/L vs 820.9 U/L +/-111.3 U/L, P < 0.01). However, opposite changes were found in PKC and MEK inhibited groups, when compared to IP group. The phosphorylation level of p44/42 MAPK was obviously decreased, the liver tissues injured evidently, and the concentrations of serum ALT and AST (645.61 U/L +/-90.4 U/L, 678.6 U/L +/-136.5U/L and 466.2 U/L +/-82.8 U/L, 732.9 U/L +/-91.1 U/L, respectively) were significantly greater than those in IP group.</p><p><b>CONCLUSION</b>These results suggest that p44/42 MAPK pathway plays a vital role in the protection of hepatocytes in ischemic preconditioning.</p>