【Abstract】ObjectiveTo measure the expressions of Fas/FasL mRNA in normal liver, adjacent non-cancerous liver parenchyma and hepatocarcinoma, and to explore the relationship between the expressions of Fas/FasL mRNA in those tissues and the hepatocellular carcinogenesis. MethodsSemi-quantity reverse transcript-ploymerase chain reaction(QRTPCR) were performed to measure the relative quantity of the Fas and FasL mRNA expressions in normal liver (n=25), adjacent noncancerous liver parenchyma(n=40) and hepatocarcinoma(n=40). ResultsThe relative quantity of Fas and FasL mRNA expressed in normal liver, adjacent non-cancerous liver parenchyma and hepatocarcinoma were 0.792±0.039 vs 0.245±0.043，0.857±0.031 vs 0.429±0.035 and 0.473±0.047 vs 0.185±0.041, respectively. The relative quantity of Fas mRNA expression in hepatocarcinoma was lower than that of normal liver tissue and adjacent non-cancerous liver parenchyrna (P＜0.05). The relative quantity of FasL mRNA expression in hepatocarcinoma was also lower than that of normal liver tissue (P＜0.05) and adjacent non-cancerous liver parenchyma (P＜0.01), but its expression in adjacent non-cancerous liver parenchyma was higher than that of normal liver tissue (P＜0.05).ConclusionHepatorcarcinoma may escape the immune surveillance of the host, not only by means of reducing Fas expression, but also through adjacent non-cancerous liver parenchyma’s increasing expression of FasL to induce apoptosis of contact lymphocyte which highly expresses Fas.
Objective
The aim is to sort CD90+ subpopulation cells in human liver cancer cell lines and investigate efficiency of magnetic cell sorting (MACS) on sorting the liver cancer stem cells.
Methods
①Expressions of CD90. Immunohistochemical method was used to determine the expressions of CD90 in normal liver tissues in 8 cases, liver cancer and adjacent liver cancer tissues in 58 cases. ②Screened the cell lines. Huh-7, MHCC97-H, Bel-7402, and SMMC-7721 cell lines were divided into blank control group and experimental group (5.5×105 cells per hole, 1 hole), cells of the experimental group were added with 5 μL CD90–PE while cells of the blank control group were treated with 5 μL CD90–PE non fluorescent antibody. Determined the proportion of CD90+ cells in the 2 groups by flow cytometry (FCM). ③MACS. Huh-7 and MHCC97-H cell lines were labeled with magnetic beads respectively and sorted by MACS, 1 mL cell suspensionsorted by magnetic sorting (MS) was collected as CD90– group, and 1 mL PBS after MS wash was collected as CD90+ group, as well as blank control group and experimental group. Determined the proportion of CD90+ cells in 4 groups by FCM. Two times of MACS were performed in Huh-7 cells. ④Serum free culture and serum culture. Huh-7 cells were divided into serum-free culture group and serum culture group (1 hole), and proportions of CD90+ cells were determined by FCM at 1 week after culture.
Results
①The positive rate of CD90 was 0 (0/8), 65.5% (38/58), and 20.7% (12/58) in normal liver tissues, liver cancer tissues, and adjacent liver cancer tissues respectively, and the positive rate of CD90 was higher in liver cancer tissues than those of normal liver tissues (χ2=6.78, P<0.05) and adjacent liver cancer tissues (χ2=20.83, P<0.05). ②For Huh-7, MHCC97-H, SMMC-7721, and Bel7402 cell lines, the proportions of CD90+ cells in the experimental group was 0.851%, 1.090%, 2.710%, and 4.050% respectively, the proportions of CD90+ cells in the blank control group was 0.241%, 0.688%, 1.890%, and 2.080% respectively, so we chose Huh-7 and MHCC97-H cell lines to perform MACS. ③Results of MACS for Huh-7 cell line. For the first MACS, the proportions of CD90+ cells in the blank control group, experimental group, CD90– group, and CD90+ group was 0.241%, 0.851%, 0.574%, and 1.100% respectively. For the second MACS, the proportions of CD90+ cells in the blank control group, experimental group, CD90– group, and CD90+ group was 0.032%, 0.961%, 0.426%, and 9.700% respectively.
Conclusions
The normal liver tissues do not express the CD90, but the liver cancer tissues express CD90 highly. There is a few CD90+ cells in Huh-7 and MHCC97-H liver cancer cell lines. The MACS has a certain effect on improving the proportion of CD90+ cells in the cell lines. The serum-free suspension culture has no effect on enriching CD90+ cells.
Parathyroid hormone (PTH) exerts multiple effects such as regulating bone remodeling, promoting angiogenesis, etc., and it is an active factor with great application potential for bone repair. In recent years, with the development of scaffold material loading strategies and parathyroid hormone-related peptides (PTHrPs), in situ loading of PTH or PTHrPs on scaffold materials to promote bone defect healing gradually becomes possible. Based on the current status and challenges of intermittent PTH (iPTH) for bone tissue engineering, the review summarizes the in-situ application strategies of PTH and the construction of PTHrPs as well as current problems and further directions in this field, with a view to propel the clinical application of scaffold materials loaded with PTH or PTHrPs in situ.
The suprachoroidal space is a potential space between the sclera and choroid. Suprachoroidal spacedrug delivery is becoming an applicable method to the ocular posterior segment diseases. Because it targets the choroid, retinal pigment epithelium and retina with high bioavailability and safety, while maintaining low levels elsewhere in the eye. In recent years, new discoveries has been carried out in different areas of interest, such as drug delivery methods, pharmacokinetics and clinical trials. Clinical trials with suprachoroidal space injection of triamcinolone acetonide are executed with promising findings for patients with noninfectious uveitis and diabetic macular edema. Suprachoroidal space triamcinolone acetonide injectable suspension is the first and currently the only agent specifically approved for uveitic macular edema by Food and Drug Administration. Nowadays, many clinical trails with suprachoroidal space drug delivery have been explored, although there are still many risks and uncertainties. With the development of technology in the future, suprachoroidal space drug delivery appears to be a promising treatment modality for ocular posterior segment diseases.
Objective To develop the plastic nano-hydroxyapatite (nano-HA)/poly (3-hydroxybutyrate-hydroxyvalerate) polyethylene glycol(PHBV-PEG) gentamicin (GM) drug delivery system(DDS)(nano-HA/PHBV-PEG-GM-DDS) for treating osteomyelitis and find its releasing character in vivo. Methods The plastic nano-HA/PHBV- PEG-GM-DDS was prepared using nanoHAas the core carrier of GM, nano-HA with PHBV and PEG as coating and plastic fibrin glue(FG) as microsphere scaffold. The morphological features of nano-HA,drug loaded nano-HA and drug loaded nano-HA/PHBVPEG microsphere were examined by electron microscope.The GM concentration in blood, cortex bone and cancellousbone was detected at 12 different time points by the method of K-B after the plastic nano-HA/PHBV-PEGGM-DDS was implanted into the femora of 36 rabbits. Its GM releasing character was assayed in vivo. Results Nano-HA was similar to a blackjack, and its length was less than 60 nm. Drug loaded nano-HA appeared natural crystal condensate, of which surface adsorbed massive GM. The average grain diameter was 200.5 nm. Drug loaded nanoHA/PHBV-PEG microsphere had a shrinkable porous structure, of which surface configuration was consistent. The average grain diameter was 34.5 μm. The GM concentration and the antibacterial annulus was in the linear correlation. The correlation coefficient was 0.998. In cortex and cancellous bone tissue, the GM concentration was about 95.50±16.50 μg/ml and 80.20±13.80 μg/ml from the plastic nano-HA/PHBV-PEG-GM-DDS on the 1st day, then decreased gradually. After 56 days of operation, the GM concentration still exceeded the minimum inhibitory concentrationfor the staphylococcus aureus, but the peak level of serum GM concentration wasunder the nephrotoxicity concentration. Conclusion Plastic nano-HA/PHBV-PEG-GM-DDS was a good drug delivery system with sustained antibiotic effect in vivo. It was an effective method for the treatment of osteomyelitis.
Objective To discuss venous drainage types of median hepatic lobe and their guiding significances on the selection of grafts. Methods Between April 2005 and March 2009, 109 potential living donors underwent 3-dimensional reconstruction of computed tomography (CT) and the volume of graft was determined in the center of organ transplantation of Ruijin Hospital. The venous drainage types of median hepatic lobe of each donor were analyzed by the computer-based liver operation-planning system in detail to assign middle hepatic vein (MHV) types according to Marcos classification and venous types of Ⅳb segment according to Nakamura classification. Results The branching pattern of MHV was divided into 3 types: Type Ⅰ and Ⅱwere relatively more accounting for 44.0% （48/109）, 37.6% （41/109）, and type Ⅲ was fewest 〔18.3% （20/109）〕. There were no significant differences in volume of whole liver, volume of left liver or left liver/total liver volume ratio among various types of MHV of the donor (Pgt;0.05). Ⅳb vein was also divided into 3 types: The most common was type Ⅰ, accounting for 72.4% (79/109); Type Ⅱ 〔12.8% （14/109）〕, type Ⅲ 〔14.7% （16/109）〕 were relatively fewer. At last, 37 donors provided right liver, for Marcos Ⅰ, Ⅱ, and Ⅲ type of donors, donors remained with MHV was 12/17, 8/11, and 5/9; for Nakamura Ⅰ, Ⅱ, and Ⅲ type of donors, those number were 16/26, 4/6, and 5/5. Conclusion In adult-to-adult living donor liver transplantation, there may be great significances in accordance with Marcos and Nakamura typing results to harvest right lobe liver graft with or without MHV.
Objective The aim of this article is to verify the clinical effect of the near-infrared fluorescent liver cancer surgery projection navigation system without display screen. Methods Three patients who need to undergo open hepatectomy for liver cancer in the Affiliated Hospital of Southwest Medical University from March 2021 to May 2021 were included, verifying the accuracy, stability, and time delay effect of the self-developed near-infrared fluorescence projection navigation system for the location of tumor in surgeries. Results The intraoperative tumor location could be accurately displayed by the near-infrared fluorescence projection system and there was no significant difference between the location of the tumor displayed by intraoperative ultrasound. The tumor location displayed by the near-infrared fluorescence projection system was not influenced by the tumor movement and had no visual-time delay. Postoperative pathology confirmed that the projection range was consistent with the tumor range. Conclusion This near-infrared fluorescence projection technology innovates the intraoperative tumor imaging mode and can accurately navigate open hepatectomy in small sample trials, and it is expected to achieve wide clinical application through subsequent iterative optimization and verification.
ObjectiveTo expounded the relationship between phenylalanine, tyrosine and their metabolites and non-alcoholic fatty liver disease (NAFLD). MethodThe literatures related to NAFLD in recent years were reviewed and analyzed. ResultThe levels of phenylalanine, tyrosine and their metabolites had changed significantly in the occurrence and development of NAFLD, and could lead to the progress of NAFLD by affecting the related pathways of lipid metabolism. ConclusionPhenylalanine, tyrosine and their related metabolites are associated with NAFLD, but the specific pathogenesis is still unclear.
ObjectiveTo explore the protective effects of abdominal paracentesis drainage (APD) on pancreatitis-associated liver injury in the early phase of severe acute pancreatitis (SAP). MethodsOne hundred and fourteen consecutive patients with SAP, admitted to the General Hospital of Western Theater Command from January 2015 to January 2021, were included in this retrospective study. The patients were divided into the APD group (n=61) and the non-APD group (n=53) based on whether they underwent APD treatment within 72 h of admission. The variables including baseline data, liverfunction tests, inflammation indexes, severity scores and other variables of the two groups were statistically analyzed. ResultsThe hospital mortality in the APD group was lower than that in the non-APD group (8.2% vs. 22.6%, P=0.031). These severity scores (including APACHE Ⅱ score, Ranson score and modified Marshall score) and inflammation indexes (including C-reactive protein, interleukin-6, interleukin-1 and tumor necrosis factor-α) in the APD group were all lower than those in the non-APD group (P＜0.05). In terms of liver function related indexes, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (GGT), total bilirubin (TBIL), and direct bilirubin (DBIL) after treatment in both two groups were significantly lower than those before treatment (P＜0.05). The levels of ALT, AST, TBIL and DBIL after treatment in the APD group were lower than those in the non-APD group (P＜0.05), and the levels of prealbumin and albumin after treatment in the APD group were higher than those in the non-APD group (P＜0.05), but there were no significant differences in the levels of alkaline phosphatase, GGT and 5′ -nucleotidase after treatment in the two group (P＞0.05). ConclusionFor SAP patients with ascitic fluid, application of APD can attenuate liver injury and improve liver function in the early stage of SAP.
Portal vein blood flow is very important for the normal function of transplanted liver. The author reviewed the management methods of different portal vein thrombosis classification in the liver transplantation (LT). The prognosis of LT in the patients with Yerdel 1–3 thrombosis is similar to that the patients without thrombosis. The portal vein reconstruction of the patients with Yerdel 4 thrombosis can be realized by varicose vein to portal anastomosis, renoportal anastomosis or cavoportal hemitransposition. When anastomosis is made at the proximal side of a spontaneous shunt between the portal and cava system, the blood shunted from portal system can be reintroduced into the donor liver, which is crucial for the management of Yerdel 4 thrombosis. The establishments of artificial shunt by distal splenic vein, mesenteric vein or “multiple to one” anastomosis are effective attempts to drain the blood from portal system to the donor liver. For more severe diffuse thrombosis of portal vein system, multivisceral transplantation, including liver and small intestine, should be considered. The cases of LT in the patients with complex portal vein thrombosis are increasing, however the prognosis remains to be determined after accumulation of the cases.
