OBJECTIVE: To investigate the selection and identification of human keratinocyte stem cells(KSC) in vitro. METHODS: According to the characteristics of KSC which can adhere to extracellular matrix very fast, we selected 3 groups of different time(5 minutes, 20 minutes and 60 minutes) and unselected as control group. And the cells were identified by monoclone antibody of beta 1-integrin and cytokeratin 19 (Ck19), then the image analysis was done. Furthermore we analyzed the cultured cells with flow cytometer(FCM) and observed the ultrastructure of the cell by transmission electron microscope(TEM). RESULTS: The cell clones formed in all groups after 10 to 14 days, while the cells of 5 minute group grew more slowly than those of the other groups, however, the clones of this group were bigger. The expression of beta 1-integrin and Ck19 were found in all groups. The positive rate of beta 1-integrin was significant difference between 5 minute group and the other groups (P lt; 0.05). And the expression of Ck19 was no significant difference between 5 minute group and 20 minute group(P gt; 0.05), and between 60 minute group and control group. But significant difference was observed between the former and the later groups(P lt; 0.05). The result of FCM showed that most cells of the 5 minute group lied in G1 period of cell cycle, which was different from those of the other groups. At the same time, the cells of 5 minute group were smaller and contained fewer organelles than those of the other groups. CONCLUSION: The above results demonstrate that the cells of 5 minute group have a slow cell cycle, characteristics of immaturity, and behaving like clonogenic cells in vitro. The cells have the general anticipated properties for KSC. So the KSC can be selected by rapid attachment to extracellular matrix and identified by monoclone antibody of beta 1-integrin and Ck19.
Portal hypertension caused by viral post hepatic liver cirrhosis has brought a heavy burden to medical treatment in China. In addition to liver transplantation, the treatments include drugs, endoscopy, intervention and surgery, but the effect is not satisfactory. At present, the consensus and guideline for portal hypertension often focuse on a single treatment, and there is an urgent need for reasonable, standardized and individualized treatment to minimize the risk of upper gastrointestinal bleeding and improve the long-term survival of patients, which should also be the ultimate goal of multi-disciplinary treatment (MDT) mode of portal hypertension. The MDT diagnosis and treatment method of portal hypertension needs to be obtained in combination with the general situation of patients (including liver function level, upper gastrointestinal bleeding risk, hypersplenism, etc.) and local medical advantages. For patients with acute upper gastrointestinal bleeding, the treatment with less trauma and good hemostatic effect should be selected as far as possible. Patients with previous bleeding history or bleeding cessation should improve the relevant evaluation as soon as possible and submit it to MDT for discussion and selection of the next appropriate treatment. Drugs and endoscopy can be used for high-risk groups of upper gastrointestinal bleeding. Whether surgical preventive treatment can benefit patients or not needs further large sample research support. Minimally invasive surgery is the development direction of surgical treatment. Combination of internal and external treatment may give full play to their respective advantages, reducing the risk of bleeding and improving long-term survival.
Objective To investigate the effect of allogeneic chondrocytes-calcium alginate gel composite under the intervention of low intensive pulsed ultrasound (LIPUS) for repairing rabbit articular cartilage defects. Methods Bilateral knee articular cartilage were harvested from 8 2-week-old New Zealand white rabbits to separate the chondrocytes by mechanical-collagen type II enzyme digestion. The 3rd passage chondrocytes were diluted by 1.2% sodium alginate to 5 × 106 cells/mL, then mixed with CaCl2 solution to prepare chondrocytes-calcium alginate gel composite, which was treated with LIPUS for 3 days (F0: 1 MHz; PRF: 1 kHz; Amp: 60 mW/cm2; Cycle: 50; Time: 20 minutes). An articular cartilage defect of 3 mm in diameter and 3 mm in thickness was established in both knees of 18 New Zealand white rabbits (aged 28-35 weeks; weighing, 2.1-2.8 kg), and divided into 3 groups randomly, 6 rabbits in each group: LIPUS group, common group, and model group. Defect was repaired with LIPUS-intervention gel composite, non LIPUS-intervention gel composite in LIPUS group and common group, respectively; defect was not treated in the model group. The general condition of rabbits was observed after operation. The repair effect was evaluated by gross and histological observations, immunohistochemical staining, and Wakitani score at 8 and 12 weeks after operation. Results Defect was filled with hyaline chondroid tissue and white chondroid tissue in LIPUS and common groups, respectively. LIPUS group was better than common group in the surface smooth degree and the degree of integration with surrounding tissue. Defect was repaired slowly, and the new tissue had poor elasticity in model group. Histological observation and Wakitani score showed that LIPUS group had better repair than common group at 8 and 12 weeks after operation; the repair effect of the 2 groups was significantly better than that of model group (P lt; 0.05); and significant differences in repair effect were found between at 8 and 12 weeks in LIPUS and common groups (P lt; 0.05). The collagen type II positive expression area and absorbance (A) value of LIPUS and common groups were significantly higher than those of model group (P lt; 0.05) at 8 and 12 weeks after operation, and the expression of LIPUS group was superior to that of common group at 12 weeks (P lt; 0.05); and significant differences were found between at 8 and 12 weeks in LIPUS group (P lt; 0.05), but no significant difference between 2 time points in common and model groups (P gt; 0.05). Conclusion Allogeneic chondrocytes-calcium alginate gel composite can effectively repair articular cartilage defect. The effect of LIPUS optimized allogeneic chondrocytes-calcium alginate gel composite is better.
Objective To investigate the clinical characteristics and prognosis of antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis with acute kidney injury (AKI) as the first manifestation, and provide new ideas for the prevention and treatment of this disease. Methods A retrospective analysis was performed on 144 patients diagnosed with ANCA-associated vasculitis in Affiliated Hospital of Southwest Medical University between August 2013 and March 2020. The patients were divided into AKI group and non-AKI group according to whether they were complicated with AKI at admission, and the differences in clinical characteristics were analyzed. The risk factors were screened by multiple logistic regression analysis. Results Among the 144 patients with ANCA-associated vasculitis, 30 cases (20.8%) were complicated with AKI at admission, and 70 cases (48.6%) died by the end of follow-up. There were 16 death cases (53.3%) in the AKI group, and 54 death cases (47.4%) in the non-AKI group, but the difference was not statistically significant (P>0.05). Single-factor analyses showed that in the AKI group, the pre-admission incidence of hematuria, neutrophil count, serum creatinine, systolic blood pressure, and Birmingham Vasculitis Activity Score were higher than those in the non-AKI group, while the red blood cell count and estimated glomerular filtration rate (eGFR) were lower than those in the non-AKI group, and the differences were statistically significant (P<0.05). Multiple logistic regression analysis showed that the neutrophil count [odds ratio (OR)=1.172, 95% confidence interval (CI) (1.003, 1.371), P=0.046] and eGFR [OR=0.942, 95%CI (0.907, 0.979), P=0.002] were independent influencing factors for AKI. Conclusions Elevated neutrophil count is an independent risk factor for ANCA-associated vasculitis complicated with AKI. It has certain guiding significance for clinical work. Early identification and intervention of these patients may contribute to reduce the case fatality rate and improve prognosis.
【Abstract】ObjectiveTo prospectively study the effects of recombinant human growth hormone (rhGH) on the changes of liver function and nutritional metabolism in postoperative patients with cirrhosis and portal hypertension. MethodsFortyeight cases with liver cirrhosis and portal hypertension who were collected from February 2003 to January 2004 were randomly divided into 2 groups (24 patients in each group). All patients were given the low calorie parenteral nutrition support and exogenous albumen after operations. Patients in the study group received rhGH from the second day after operations and physiological saline was used in the control group instead. The effects were evaluated in terms of protein metabolism, liver function, blood glucose level at different phases before and after the intervene. Death rates of in patients were also recorded in both groups. ResultsThe rising amplitude of albumen in the study group had been significantly larger than that of the control group from the seventh day after intervene (P＜0.05). The blood transaminase levels (ALT，AST) in the study group were significantly lower than that of the control group (P＜0.05). The blood glucose level of both groups decreased over time and returned to normal on day 14 after intervene, but there was no significant difference for both glucose and plasma bilirubin level between the two groups before and after the intervene (Pgt;0.05). The rates of death were similar, although the length of stay in the study group was much shorter than that of the control group. ConclusionrhGH may inhibit the catabolism, correct hypoproteinemia, improve liver function for postoperative patients with cirrhosis and portal hypertension, and reduce their length of stay.