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Objective:To compare the prognoses of salvage liver transplantation fulfilling the Criteria of Milan, University of California San Francisco(UCSF)and Hangzhou.Methods:Clinical data were retrospectively reviewed for 256 patients with recurrent hepatocellular carcinoma(HCC)undergoing donation after citizen death(DCD)liver transplantation(LT)from January 2015 to October 2019.They were divided into two groups of primary(PLT, n=175)and salvage(SLT, n=81). General profiles, tumor pathological characteristics and postoperative complications of two groups were compared by T-test, rank-sum or χ2 test.Kaplan-Meier method and Log rank test were employed for comparing overall survival rate(OS)and recurrence-free survival rate(RFS)between two groups.In SLT group, 31 cases fulfilled Milan criteria, 45 cases UCSF criteria and 69 cases Hangzhou criteria.OS/RFS of three groups were compared.According to there was downstaging or bridging treatment pre-LT, SLT group was divided into downstaging group(n=32)and non-downstaging group(n=49). OS/RFS of two groups were compared.According to the Rescit1.1 criteria, downstaging group were divided into remission group(n=14)and non-remission group(n=18)and OS/RFS of two groups were compared. Results:The operative durations of PLT and SLT groups were(439.5±74.9)and(475.1±83.4)min respectively.There was significant inter-group difference( P<0.05); However, no significant inter-group difference existed in amount of intraoperative bleeding, blood transfusion, postoperative hospital stay or incidence of postoperative complications(all P>0.05). No significant difference existed in OS/RFS between PLT and SLT groups( P>0.05). No significant difference existed in OS at 1/3/5 years post-SLT among Milan, UCSF and Hangzhou criteria groups(all P>0.05); However, RFS in Milan criteria group at 1/3/5 years post-SLT were 93.5%, 81.7% and 81.7% respectively.They were significantly higher than 68.9%, 59.7% and 59.7% in UCSF criteria group and 78.3%, 58.8% and 55.5% in Hangzhou criteria group(all P<0.05). For patients on downstaging therapy, OS in the Remission group at 1, 3 and 5 years post-SLT were 100%, 73% and 73% respectively, which was significantly higher than 83.3%, 49.4% and 0 in non-Remission group( P=0.042). RFS in the Remission group at 1, 3 and 5 years post-SLT were 100%, 62.5% and 46.9% respectively, which was significantly higher than 52.9%, 0 and 0 in no-Remission group( P=0.001). Conclusions:The survival outcome of SLT recipients is similar to that of PLT recipients.The overall survival of SLT recipients shows no significant difference between Milan, UCSF and Hangzhou criteria.However, SLT recipients fulfilling Milan criteria have the longest recurrence-free time.The prognosis of patients with remission after preoperative descending treatment is superior to that of patients without remission.
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Micropatterning is an established technique in the cell biology community used to study connections between the morphology and function of cellular compartments while circumventing complications arising from natural cell-to-cell variations. To standardize cell shape, cells are either confined in 3D molds or controlled for adhesive geometry through adhesive islands. However, traditional micropatterning techniques based on photolithography and deep UV etching heavily depend on clean rooms or specialized equipment. Here we present an infrared laser assisted micropatterning technique (microphotopatterning) modified from Doyle et al. that can be conveniently set up with commercially available imaging systems. In this protocol, we use a Nikon A1R MP+ imaging system to generate micropatterns with micron precision through an infrared (IR) laser that ablates preset regions on poly-vinyl alcohol coated coverslips. We employ a custom script to enable automated pattern fabrication with high efficiency and accuracy in systems not equipped with a hardware autofocus. We show that this IR laser assisted micropatterning (microphotopatterning) protocol results in defined patterns to which cells attach exclusively and take on the desired shape. Furthermore, data from a large number of cells can be averaged due to the standardization of cell shape. Patterns generated with this protocol, combined with high resolution imaging and quantitative analysis, can be used for relatively high throughput screens to identify molecular players mediating the link between form and function.
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Rayos Láser , Alcohol PolivinílicoRESUMEN
Objective:To explore the relationship between CD24 expression in preoperative peripheral blood as well as cancer tissue and clinical parameters and prognosis in patients with hepatocellular carcinoma (HCC) after liver transplantation (LT).Methods:From November 2018 to November 2019, clinical data were collected for 65 HCC patients and 41 patients with benign liver disease.The preoperative peripheral blood level of CD24 was detected by enzyme-linked immunosorbent assay (ELISA) and the expression of CD24 in cancerous foci and adjacent tissues examined by immunohistochemistry.Kaplan-Meier survival curves of differential CD24 expression were plotted and survival differences compared by Log-rank method.One-way ANOVA was utilized for examining the relationship between the expression level of CD24 and various clinicopathological parameters and multivariate Cox analysis for screening independent risk factors affecting patient prognosis.Results:The concentration of CD24 in preoperative peripheral blood (p-CD24) of HCC patients (6.51±2.33 μg/L) was significantly higher than that of patients with benign liver disease (4.10±0.91) μg/L, P<0.05.The positive rate of CD24 was obviously higher in cancerous tissues than that in adjacent tissues (87.7% vs. 4.6%, P<0.05). The peripheral blood level of CD24 was positively correlated with the expression intensity of CD24 in tumor tissues (t-CD24, r=0.570, P<0.001). The expression of CD24 (both in blood and cancer foci) was significantly correlated with preoperative level of gamma-glutamyl transferase (GGT), maximal tumor diameter, microvascular invasion, portal vein tumor thrombus, vessel carcinoma embolus and satellite focus ( P<0.05). The expression of CD24 in patients exceeding the Milan/UCSF criteria was higher than those fulfilling the criteria ( P<0.005). Patients with a higher expression of CD24 had worse overall survival and recurrence-free survival rates as compared to those a lower expression of CD24 ( P<0.05). Multivariate Cox analysis indicated that t-CD24 [OS: HR=3.661(1.005-13.333)], P=0.049; recurrence-free survival (RFS): [HR=4.331(1.887-9.942), P=0.001] and preoperative level of alpha fetoprotein (AFP) [OS: HR=4.900(1.590-15.097), P=0.006]; RFS: [HR=3.414(1.614-7.221), P=0.001] were independent risk factors for overall survival and recurrence-free survival in HCC patients undergoing LT. Conclusions:The preoperative peripheral blood level of CD24 in HCC patients undergoing LT indirectly reflects the expression of CD24 in cancerous tissues to a certain extent.And the expression of CD24 in cancerous tissue is one of the independent risk factors affecting OS and RFS of LT patients.
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Objective To summarize the screening Methods for human parvovirus (HPV) B19 infection after liver transplantation and analyze the related risk factors. Methods Clinical data of 86 recipients were retrospectively analyzed. According to the Results of next generation sequencing (NGS), all recipients were divided into the HPV B19 infection group and control group. Clinical characteristics, treatment regime and clinical prognosis of patients infected with HPV B19 were analyzed. The risk factors of HPV B19 infection were analyzed using univariate and multivariate Logistic regression model by forward LR step method. Results Nine of the 86 recipients developed fever and progressive anemia with unexplained reasons at approximately 2 weeks after liver transplantation. NGS detection demonstrated that HPV B19 was positive and they were diagnosed with pure red cell aplasia (PRCA) caused by HPV B19 infection. After intravenous immunoglobulins (IVIG) was given and the immunosuppressant therapy was adjusted, the hemoglobin levels in all patients were significantly increased. The Results of multivariate analysis revealed that low serum globulin level in peripheral blood at postoperative 7 d [odds ratio (OR) =0.749, P=0.040] and young age (OR=0.937, P=0.038) were the independent risk factors of HPV B19 infection after liver transplantation. Conclusions HPV B19 infection should be considered in relatively young patients with unexplained hemoglobin decline early after liver transplantation. NGS screening is an effective method for early diagnosis of HPV B19 infection. Low serum globulin level in peripheral blood at postoperative 7 d and young age may be independent risk factors of the incidence of HPV B19 infection.
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Objective To investigate the community structure of intestinal bacteria from patients with cirrhosis and its influencing factors. Methods From 2016 to 2017, 24 patients with liver cirrhosis ( the LC group) and 23 healthy family members of patients ( the HC group) were enrolled at the First Hospital of Lanzhou University. A comparative analysis of the community structure of intestinal bacteria was performed using 16S rRNA gene sequencing in LC and HC groups. Combined with LEfSe analysis and NMDS analysis, the differential markers were screened and the factors affecting the intestinal community structure of subjects were studied. Results The dominant six phylum of bacteria in intestines in LC and HC groups included Firmicutes, Bacteroides, Proteobacteria, Actinobacteria, Fusobacteria and Tenericumes. However, in the LC sample, Firmicutes was significantly reduced, while Bacteroides was significantly increased. The diversity of intestinal bacteria was significantly reduced, and the Firmicutes/Bacteroides ratio was significantly decreased, suggesting a variation of the community structure in intestinal bacteria of cirrhosis patients. The LEfSe result indicated that the abundance of Enterococcus, Lactobacillales, Bacilli, and Bacteroidetes showed a significant difference in the LC sample, which may be used as potential marked bacterial groups for cirrhosis. The NMDS analysis revealed a positive relationship between the concentration of Cd and Pb and the abundance of intestinal bacteria in the LC sample. Conclusion The community structure of intestinal bacteria from patients with cirrhosis has changed. Enterococcus, Lactobacillales, Bacilli, and Bacteroidetes are potential marked bacterial groups. The concentration of Cd and Pb in the intestinal tract of cirrhosis patients may interact with the abundance and structure of bacteria, and further affect the occurrence and development of cirrhosis.
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Objective To explore the clinical efficacies of applying aged marginal donor liver . Methods From January 2015 to June 2018 ,clinical data were retrospectively analyzed for 199 adult liver transplantation donors and recipients .They were divided into two groups of aged (≥60 years) and appropriate age (<60 years) .The prognosis of two groups was compared after a follow-up period of 1 year .And the aged group was further assigned into lower and higher fat infiltration groups according to the degree of fat infiltration in donor liver and compared the prognosis of two groups . Results No significant differences existed in initial ,peak value and recovery time of transaminase (AST/ALT ) , peak value and recovery time of total bilirubin , glutamyl transpeptidase , alkaline phosphatase ,international normalized ratio (INR) ,peak value of lactate ,postoperative hospital stay , graft dysfunction , biliary/vascular complications , acute/chronic rejection or graft survival rate between aged and appropriate age groups post-transplantation .The aged group was further divided into lower and higher fat infiltration groups according to the fat infiltration rate (< 20% ,≥20% ) . And significant inter-group differences existed in peak value and recovery time of AST /ALT ,peak value of total bilirubin , glutamyl transpeptidase , lactate , postoperative hospital stay and graft dysfunction .The above parameters were significantly worse in higher fat infiltration group .Also the rejection rate was higher in high group at 1 year post-operation and no significant inter-group difference existed in biliary/vascular complications . In higher group , 4 patients showed graft dysfunctions during perioperative period .Two of them were discharged successfully after secondary transplantation and another 2 patients died .Conclusions On the premise of comprehensive evaluations of donor liver status and reasonable matching of recipients ,aged marginal donor liver can be safely applied with excellent clinical outcomes .Severe fatty donor liver should be employed with caution . Hypertensive drugs ,high serum sodium and long period of cold ischemia are also important influencing factors for aged donors .
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Objective@#To explore the clinical efficacies of applying aged marginal donor liver.@*Methods@#From January 2015 to June 2018, clinical data were retrospectively analyzed for 199 adult liver transplantation donors and recipients. They were divided into two groups of aged (≥60 years) and appropriate age (<60 years). The prognosis of two groups was compared after a follow-up period of 1 year. And the aged group was further assigned into lower and higher fat infiltration groups according to the degree of fat infiltration in donor liver and compared the prognosis of two groups.@*Results@#No significant differences existed in initial, peak value and recovery time of transaminase (AST/ALT), peak value and recovery time of total bilirubin, glutamyl transpeptidase, alkaline phosphatase, international normalized ratio (INR), peak value of lactate, postoperative hospital stay, graft dysfunction, biliary/vascular complications, acute/chronic rejection or graft survival rate between aged and appropriate age groups post-transplantation. The aged group was further divided into lower and higher fat infiltration groups according to the fat infiltration rate (<20%, ≥20%). And significant inter-group differences existed in peak value and recovery time of AST/ALT, peak value of total bilirubin, glutamyl transpeptidase, lactate, postoperative hospital stay and graft dysfunction. The above parameters were significantly worse in higher fat infiltration group. Also the rejection rate was higher in high group at 1 year post-operation and no significant inter-group difference existed in biliary/vascular complications. In higher group, 4 patients showed graft dysfunctions during perioperative period. Two of them were discharged successfully after secondary transplantation and another 2 patients died.@*Conclusions@#On the premise of comprehensive evaluations of donor liver status and reasonable matching of recipients, aged marginal donor liver can be safely applied with excellent clinical outcomes. Severe fatty donor liver should be employed with caution. Hypertensive drugs, high serum sodium and long period of cold ischemia are also important influencing factors for aged donors.
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OBJECTIVE: To determine whether the immunopotentiator chitosan oligosaccharide(COS)can recover the reproductive functions of pathological ovarian recession mice and improvetheir immunity. METHODS: Forty-three fertile female mice (at around 2 months),in addition to a normal control group (n=8), injected intraperitoneally with busulfan and cyclophosphamide to construct premature ovarian failure mod-els. Three of them were used to test whether the infertility model was constructed successfully by HE staining. Then the models were randomly divided into four groups (n=8) and treated with different dosages of COS by gavage, after which compared different groups' organ ratios (the weight of immune organs and ovary/body weight), ovarian follicles and peritoneal macrophages' phagocytosis as well as estragon(E2) and pro-gesterone(P) levels in peripheral blood. In addition, we measured the expression dynamics of the ovarian protein reproductive cell marker mouse vasa homolog(MVH), germ stem cell marker OCT-4 in ovarian surface epithelium (OSE) and part of immune factors including tumor necrosis factor (TNF-α),interleukin-2(IL-2)as well as IL-6 to analyze the correlativity between germline stem cells marker dynamics and im-mune factors expression changes. RESULTS: With increasing dosages of COS, organ ratios of ovaries, thymus and spleen both went up syn-chronously; The whole number of follicles and every stages of follicles are all presenting with progressive tendency; E2 level in peripheral blood ascends, however, progesterone level declined relatively; Neutral red experiment revealed the phagocytosis ability of peritoneal macrophages became stronger with increasing dosages of COS; the results of Western blot had shown that no matter the expression level of germ stem cells marker or immune factors were all presenting increasing tendency, which means that the expression level dynamics of germ stem cell marker has a positive correlation with immune factors expression changes. The results were statistically significant. CONCLUSIONS: COS could improve the immunity of mice with pathological ovarian recession and at the same time it would promote the proliferation and differentiation of female germ line stem cells (FGSCs), and then helped saving ovarian functionsto some extent.
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Envejecimiento/inmunología , Quitosano/farmacología , Células Germinativas/efectos de los fármacos , Ovario/efectos de los fármacos , Animales , Femenino , Interleucina-2/inmunología , Interleucina-6/inmunología , Ratones , Folículo Ovárico/efectos de los fármacos , Folículo Ovárico/fisiopatología , Ovario/fisiopatología , Insuficiencia Ovárica Primaria/inducido químicamente , Células Madre/efectos de los fármacos , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Objective To investigate the effect of immature dendritic cell (imDC) derived from donor-derived bone marrow induced by alkaloid sinomenine (SN) on the Th1/Th2 cytokines in venous blood of receptors,and to probe into the mechanism which imDC induced by SN can lead transplantation immune tolerance.Methods Inbred strain Wistar and Sprague Dawley (SD) rats were selected as kidney transplant donor and recipient,respectively.Vessel sutures of the microsurgery technique were used to build the bilaterally renal transplantation model of rats.By the injection of imDC to the recipient rats preoperatively,enzyme-linked immunosorbent assay (ELISA) was used to determine the level of the interleukin (IL)-2,IL-4,IL-10 and interferon-γ (INF-γ).Results (1) Successful rate of transplantation was 89.5%.Arterial anastomosis time was (12.5 ±5.7)min,and venous anastomosis time was (17.3 ± 3.4)min.(2) The content of the IL-2,INF-γ,IL-4 and IL-10 in SN-imDC 106 group was (17.25 ± 3.41) pg/ml,(239.80 ± 9.06) pg/ml,(337.60 ± 25.07) pg/ml,and (1 432.00 ± 106.39) pg/ml,respectively.Among the same concentration,the level of the IL-4 and IL-10 that stood for the Th2 cytokines was significantly higher in SN-imDC group than imDC group and control group (P < 0.05),and was significantly higher in SN-imDC 106 group than SN-imDC 105 group (P < 0.05).Among the same concentration,the lever of IL-2 and INF-γ that stood for the Th1 cytokines was significantly lower in SN-imDC group than imDC group and control group (P < 0.05),and was significantly lower in SN-imDC 106 group than SN-imDC 105 group (P < 0.05).Conclusions (1) The use of microsurgery for anastomosis could make the model of singel kidney transplantation in rats.(2) Specific imDC induced by SN could induce the migration to Th2 immune response,which proved imDC induced by SN could mediate immune tolerance to the recipient.