Objective To explore the proper dosage of establishment of stable hepatic oval cells （HOC） prolif-eration model by using 2-acetaminofluorene （2-AAF） combined with two-third partial hepatectomy （2/3 PH） surgery， and to explore isolated and cultured method of HOC in vitro. Methods The 174 Wistar rats were randomly divided into 4 experimental groups （each group enrolled 30 rats）， saline group （n=30）， and untreated group （n=24）. Rats of 4 experi-mental groups were underwent gavage of 5， 10， 15， and 20 mg/（kg ? d） 2-AAF， corresponding to the groups from No.1 to No.4 group. Rats of saline group received saline gavage and rats of untreated group didn’t received any treatment. A standard 2/3 PH surgery was performed on the 5th day after gavage， then the same gavage method was still administrated as preoperation untill rats were sacrificed. The liver tissues of 6 selected rats were adopted and identified by HE staining and immunohistochemical staining on 4， 8， 12， and 16 days after PH for observation of the proliferation of HOC in every group， on 4 days， levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） were tested in addition. HOC were isolated and purified by collagenase perfusion method and percoll gradient centrifugation. Results The surv-ival rates of untreated group，saline group，No.1 group，No.2 group，No.3 group，and No.4 group were 100% （24/24），93% （28/30），93% （28/30），90% （27/30），90% （27/30），and 80% （24/30） respectively. Compared with the saline group and untreated group， the levels of serum ALT and AST increased significantly in No.2， No.3， and No.4 group on the 4th day after PH （P＜0.05）. The results of HE staining showed that No.2， No.3， and No.4 group were observed visibly different level of damage at liver tissue， and the proliferation level of HOC were most obviously in No.3 and No.4 group. The results of immunohistochemical staining revealed that proliferation cells were positively expressed oval cell marker-6 （OV-6）. The number of OV-6 positive cells were increased significantly with the increase of dosage of 2-AAF between 4 days and 12 days after operation， and proliferation levels were related with dosages of 2-AAF （P＜0.05）. In all cultured cells， 80% of cells were OV-6 positive cells after isolation and culture by using collagenase perfusion method and percoll gradient centrifugation. Conclusions The methods of gavage of 2-AAF at 15 mg/（kg ? d） combined with 2/3 PH surgery can establish the HOC proliferation model on the 12th day， as well as the rats have lower mortality and better tolerance， especially. The collagenase perfusion method and percoll gradient centrifugation can be used to isolate HOC effectively.
The incidence of cardiovascular disease remains high, and surgery is an important measure for the treatment of cardiovascular disease. However, cardiovascular surgery is complicated and difficult, and it is one of the departments with the highest rate of allogeneic blood transfusion. Allogeneic blood transfusion significantly increases the complications and mortality of patients, while autologous blood transfusion can effectively reduce allogeneic blood transfusion and adverse reactions. Autologous plateletpheresis technology is a popular autotransfusion method in recent years. This article reviews the autologous plateletpheresis technology and its clinical application in cardiovascular surgery.
ObjectiveTo systematically review the effects of core training for diastasis recti abdominis (DRA) in postpartum period. MethodsThe PubMed, EMbase, EBSCO, Cochrane Library, CNKI, CBM and WanFang Data databases were electronically searched to collect randomized controlled trials (RCTs) on core training for patients with DRA postpartum from inception to December 7, 2022. Two reviewers independently screened the literature, extracted data, and assessed the risk of bias of the included studies. Meta-analysis was then performed using RevMan 5.4 software. ResultsA total of 12 RCTs were included, involving 741 patients with DRA postpartum. The results of the meta-analysis demonstrated that core training significantly reduced inter-recti distance (IRD) above the umbilicus (SMD=−1.37, 95%CI −2.30 to −0.44, P<0.05), below the umbilicus (SMD=−0.82, 95%CI −1.28 to −0.36, P<0.05), at the level of the umbilicus during contraction of the rectus abdominis (RA) (SMD=−0.76, 95%CI −1.24 to −0.28, P<0.05) and above the umbilicus during RA contraction (SMD=−3.41, 95%CI −5.12 to −1.69, P<0.05) in patients with DRA postpartum. Additionally, the results indicated that core training could improve visual analogue scale, abdominal circumference, waist-hip ratio, lumbopelvic control impairment, lumbopelvic proprioception, the static and dynamic overall balance stability, the static and dynamic anterior-posterior balance stability, medial-lateral static balance stability and Oswestry disability index in patients with DRA postpartum (P<0.05). However, no significant improvement was observed in inter-recti distance (IRD) below the umbilicus during RA contraction, the score of inventory of functional status after childbirth questionnaire, the score of multidimensional body-self relations questionnaire, medial-lateral dynamic balance stability or the score of pelvic floor impact questionnaire in patients with DRA postpartum (P>0.05). ConclusionCore training may improve IRD, pain intensity, total abdominal fat and fat distribution and balance in patients with DRA postpartum, but its efficacy in improving postpartum functional status, body image satisfaction or the degree of dysfunction is unclear. Due to the limited quality and quantity of the included studies, more high quality studies are required to verify the above conclusion.
Objective To review the research progress of pubic symphysis diastasis and provide effective reference for orthopedic surgeons in the diagnosis and treatment of pubic symphysis diastasis. MethodsThe anatomy, injury mechanism, treatment, and other aspects of pubic symphysis diastasis were summarized and analyzed by reviewing the relevant research literature at domestically and internationally in recent years. ResultsThe incidence of pubic symphysis diastasis is high in pelvic fractures, which is caused by the injury of the ligaments and fibrocartilage disc around the pubic symphysis by external force. The treatment plan should be individualized according to the pelvic stability and the needs of patients, aiming to restore the stability and integrity of the pelvis and improve the quality of life of patients after surgery. ConclusionAt present, the research on pubic symphysis diastasis still needs to be improved. In the future, high-quality, multi-center, and large-sample studies are of great significance for the selection of treatment methods and the evaluation of effectiveness for patients with pubic symphysis diastasis.
ObjectiveTo obtain the mesenchymal stem cells (MSCs) from human umbilical cord and mark in vitro, for further transplantation therapy. MethodsThe MSC were isolated from human umbilical cord by tissue explants culture method. After subculture in vitro, the morphology of hUC-MSC was observed; the surface antigens of hUC-MSC were detected by flow cytometry; adipogenic and osteogenic differentiation were determined by specific staining; hUC-MSC labelled with Brd U were identified by immunofluorescence. ResultsMSC could be isolated successfully by tissue explants culture method. When cultured about one week, the cells climbed out from the tissue block edge, proliferated and formed colonies; the hUC-MSCs of passage 5 were detected by flow cytometry, and they highly expressed CD73, CD90 and CD105, didn't express or lowly expressed CD14, CD34, CD45, CD79a and human leukocyte antigen-DR. After two weeks of adipogenic induction, they were positive in oil red O staining, and after three weeks of osteogenic induction, red precipitate could be seen by alizarin red staining, and the red fluorescence of the hUC-MSC labelled with Brd U could be detected by immunofluorescence detection. ConclusionThe cells can be isolated from human umbilical cord by tissue explants culture method, with the characteristics of hUC-MSCs and can be labeled successfully in vitro, so it can be used for the research in the field of cell transplantation.
Kidney transplantation is an ideal treatment for patients with end-stage renal disease. Circulating alloantibodies against donor human leukocyte antigens and blood group antigens can impair allografts, shorten allograft survival, and limit access to kidney transplantation. Furthermore, the presence of donor specific antibodies is associated with increased incidence of antibody-mediated rejection and decreased graft survival following transplantation. Plasmapheresis, an extracorporeal therapy directed at removing plasma proteins that has been found to minimize the effects of perioperative sensitization in kidney transplantation. Plasmapheresis enables transplantation across the barrier of ABO blood group incompatibility. In addition, it is also an important approach for the treatment of antibody-mediated rejection. Therefore, studying the application of plasmapheresis in perioperative period of kidney transplantation is expected to increase the chance of transplantation and improve the outcomes following transplantation. This article introduces the application of plasmapheresis in the perioperative period of kidney transplantation.
Objective To explore a simple, effective and stable method for the isolation and purification of Kupffer cells from rat liver, enabling further study on the structure and function of these cells in vitro. Methods After laparotomy, a catheter was inserted into the portal vein and secured with artery clamp. Then, the rat liver was perfused and digested with solution Ⅰ and solution Ⅱ containing 0.05% collagenase Ⅳ respectively. The cell suspension was centrifuged with isopycnic sedimentation in a two-step Percoll gradient to harvest Kupffer cells. The isolated Kupffer cells were purified by selective adherence after 30 min of cultivation, and identified by evaluation of phagocytosis of India ink and peroxidase staining with DAB through light and electron microscopy. Results It was verified that the viability of isolated Kupfffer cells was more than 90% through Trypan blue staining. Those Kupffer cells could attach to plastic quickly and phagocytose ink, and had the appearance of “fried eggs” in positive peroxidase staining with a purity of 95%. Under the light microscopy, the appearance of newly isolated Kupffer cells was round with uniform shape and size. After two days of culture, Kupffer cells appeared to distend with irregular or stellate shape. The typical features were observed in the transmission electron micrographs. There were numerous pseudopods and occasional cup-like indentations in the cell membrane of Kupffer cells. The cytoplasm contained numerous types of lysosomes and other phagocytotic vesicles. Conclusion The method for isolating and culturing Kupffer cells in this study is effective and stable, and the biological characters are preserved in the cultured cells.
ObjectiveTo compare the effectiveness of flexible fixation and rigid fixation in the treatment of ankle pronation-external rotation fractures with distal tibiofibular syndesmosis.MethodsA retrospective analysis was made on the clinical data of 50 patients with ankle pronation-external rotation fractures and distal tibiofibular syndesmosis treated between January 2013 and December 2015. Suture-button fixation was used in 23 patients (flexible fixation group) and cortical screw fixation in 27 patients (rigid fixation group). There was no significant difference in age, gender, weight, side, fracture type, and time from trauma to surgery between 2 groups (P>0.05). The operation time, medial clear space (MCS), tibiofibular clear space (TFCS), tibiofibular overlap (TFO), American Orthopaedic Foot and Ankle Society (AOFAS) score, and Foot and Ankle Disability Index (FADI) score were compared between 2 groups.ResultsThe operation time was (83.0±9.1) minutes in the flexible fixation group and was (79.6±13.1) minutes in the rigid fixation group, showing no significant difference (t=1.052, P=0.265). All patients achieved healing of incision by first intention. The patients were followed up 12-20 months (mean, 14 months). The X-ray films showed good healing of fracture in 2 groups. There was no screw fracture, delayed union or nounion. The fracture healing time was (12.1±2.5) months in the flexible fixation group and was (11.3±3.2) months in the rigid fixation group, showing no significant difference between 2 groups (t=1.024, P=0.192). Reduction loss occurred after removal of screw in 2 cases of the rigid fixation group. At last follow-up, there was no significant difference in MCS, TFCS, TFO, AOFAS score and FADI score between 2 groups (P>0.05).ConclusionSuture-button fixation has similar effectiveness to screw fixation in ankle function and imaging findings, and flexible fixation has lower risk of reduction loss of distal tibiofibular syndesmosis than rigid fixation.
Objective To review the general approaches in isolation and purification of pancreatic islets and progress in several aspects. Methods The latest l iterature concerning acquisition of pancreatic islets was reviewed and analyzed interms of the choice of pancreatic islet donors, the digestion and isolation of pancreas, the purification of islet and the assay of outcome. Results The profile of the isolation and purification depends on the selection of reagents and methods of operation in every step and l inkup between every step. Conclusion Pancreatic islet transplantation is the most effective method to treat type 1 diabetes, the problem of inadequate sources of pancreatic islets could be resolved by the optimal process and the establ ishment of standardized operation.