Objective To investigate the effect of the serum from severe burn patients on the biology characteristics of human umbilical cord mesenchymal stem cells (hUCMSCs) in vitro, so as to explore the feasibility of hUCMSCs transplantation for treating severe burn. Methods The 3rd passage of hUCMSCs were randomly divided into 3 groups: 10% fetal bovine serum group (group A), 10% normal serum group (group B), and 10% burn serum group (group C). At 24 hours, 72 hours, and 6 days after culture, the cell morphology and density were observed by inverted microscope; the cell proliferation was assessed by MTT; after 6 days of culture, the cell cycle by propidium iodide staining and flow cytometry, the apoptosis by acridine orange/ethidium bromide staining, and the cell senescence by β-galactosidase staining; the levels of tumor necrosis factor α (TNF-α), interleukin 1 (IL-1), platelet-derived growth factor (PDGF), and insulin-like growth factor 1 (IGF-1) in serum were detected by a double-antibody sandwich ELISA kit. Results hUCMSCs were long spindle/polygon in 3 groups. The cell fusion of group C was obviously faster than that in group A and group B. The cell proliferation curves showed that the velocity and number of cell proliferation in group C were significantly higher than those in group A and group B at 2-6 days after culture (P lt; 0.05). The rates of proliferation period (S) of hUCMSCs were 9.21% ± 1.02%, 11.79% ± 1.87%, and 20.54% ± 2.03%, respectively in groups A, B, and C at 6 days, and group C was significantly higher than that of group A and group B (P lt; 0.05). The hUCMSCs showed normal morphology and structure in 3 groups, and no apoptosis cells was observed. The positive cells percentage of group C (2.6% ± 0.1%) was significantly lower than that of group A (4.8% ± 0.2%) and group B (3.8% ± 0.4%) (P lt; 0.05). The levels of TNF-α, IL-1, PDGF, and IGF-1 in group C were significantly higher than those in group B (P lt; 0.05). Conclusion The higher levels of cytokines in serum from the severe burn patients can significantly stimulate hUCMSCs proliferation, prevent cells apoptosis, and reduce cells senescence. Therefore, it is feasible to use hUCMSCs transplantation for treating severe burn patients.
ObjectiveTo summarize the current research progress of serum exosome microRNAs in patients with colorectal cancer.MethodsThe domestic and foreign literatures related to serum exosome microRNAs of colorectal cancer patients, which had been reported in recent years were collected through literature search. Subsequently, those literatures were used to read and review.ResultsExosomes were extracellular vesicles, which contained lipids, proteins, DNA, RNA (mRNA, microRNA, and long non-coding RNA), and other molecules. These vesicles mediated communication between cells by transporting the above molecules. Exosomes in serum were the main carriers of microRNAs in the blood circulation system. Serum exosome microRNAs could affect the proliferation, invasion, and metastasis of colorectal cancer cells, mediate the drug resistance of colorectal cancer cells, and could be used as biomarkers to predict the prognosis of colorectal cancer.ConclusionsSerum exosome microRNAs play important role in the occurrence, development, treatment, and diagnosis of colorectal cancer. As a class of biomarker, serum exosome microRNAs have great potential in the early diagnosis and prognostic evaluation of colorectal cancer.
ObjectiveTo investigate the effect of serum on the differentiation of neural stem cells.MethodsThe neural stem cells were isolated from the embryonic hippocampus tissues of Sprague Dawley rats at 14 day of pregnancy. After culturing and passaging, the 3rd generation cells were identified by immunocytochemical staining. Then, the cells were divided into 3 groups according to the concentrations of fetal bovine serum (FBS) used in the differentiation cell culture medium: 5% (group A), 1% (group B), 0 (group C), respectively. The other components of the culture media in 3 groups were the same. Cell viability was determined by using the Live/Dead cell staining at 8 days; the expressions of glial cell marker [glial fibrillary acidic protein (GFAP)] and neuronal marker (β-Ⅲ Tubulin) were determined and analyzed by immunocytochemical staining and real-time fluorescent PCR at 4 and 8 days of culture.ResultsBased on cell morphology and immunocytochemical staining, neural stem cells were identified. Cells were growing well with no death in all groups. With decreasing FBS concentration, the expression of GFAP was significantly decreased on both protein and mRNA level, whereas the expression of β-Ⅲ Tubulin was evidently increased. The staining of each group at 8 days was more obvious than that at 4 days. There were significant differences in mRNA expressions of GFAP and β-Ⅲ Tubulin at 4 and 8 days between groups (P<0.05).ConclusionSerum can promote the differentiation of neural stem cells into glial cells. At the same time, it inhibits the differentiation of neural stem cells into neurons, the lower the serum concentration, the smaller the effect.
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.
ObjectiveTo analyze the R0 resection rate and survival time of pancreatic cancer with serum IgG4 elevated, and to discuss whether serum IgG4 can distinguish autoimmune pancreatitis from pancreatic cancer.MethodsThe retrospective cohort study was adopted. The clinical data of 146 patients with pancreatic cancer confirmed by histology in Affiliated Hospital of Qingdao University from January 2016 to December 2019 were analyzed retrospectively. According to the level of serum IgG4, they were divided into normal IgG4 group (<1.35 g/L, n=124) and IgG4 elevated group (≥1.35 g/L, n=22). The tumor R0 resection rate, survival time and whether complicated with AIP of the two groups were compared.ResultsOne hundred and one patients (81.5%) with normal serum IgG4 underwent radical surgery, while only 13 patients (59.1%) with elevated serum IgG4 underwent radical surgery, the difference was significant (P=0.019). The median survival time of patients with normal serum IgG4 was 18.7 months, while patients with elevated serum IgG4 was 8.1 months, there was no significant difference between the two groups (P=0.121). Subgroup analysis showed that the median survival time of patients with pancreatic ductal adenocarcinoma in the normal IgG4 group was 17.5 months, while the IgG4 elevated group was 6.8 months, the difference was significant (P=0.016). Only 1 case of pancreatic cancer with AIP in the 2 groups.ConclusionsSerum IgG4 ≥1.35 g/L indicates low radical resection rate in pancreatic cancer and poor prognosis in pancreatic ductal adenocarcinoma. Serum IgG4 can only be used as an auxiliary index to distinguish pancreatic cancer from autoimmune pancreatitis.
Objectives To evaluate the expression levels of serum microRNA-21 (miRNA-21) and microRNA-155 (miRNA-155) from patients and rats with pancreatic cancer, and to explore its value in the diagnosis of pancreatic cancer. Methods The clinical materials and the serum samples from 18 patients with pancreatic cancer (pancreatic cancer group) and 12 patients with benign pancreatic disease (benign pancreatic disease group) admitted to Fujian Medical University Union Hospital between January 2016 and December 2016 were collected prospectively. The real-time fluorescent quantitative PCR was performed to detect the levels of serum miRNA-21 and miRNA-155. 7, 12-dimethylbenz (a) anthracene (DMBA)-induced pancreatic cancer rat models (n=20) and the models of the blank control group (sham operation, n=10) were established and the serum samples from the pancreatic cancer group and the blank control group were measured by the real-time fluorescent quantitative PCR, to detect the levels of miRNA-21 and miRNA-155. Results The median expression levels of serum miRNA-21 and miRNA-155 were 1.99 (1.43–5.30) and 7.06 (4.98–21.48) in the pancreatic cancer group, as well as 1.28 (0.58–2.01) and 2.20 (1.76–3.02) in the benign pancreatic disease group. The expression levels of serum miRNA-21 and miRNA-155 were significantly higher in the pancreatic cancer group (Z=–2.621,P=0.009; Z=–3.430,P=0.001). In animal studies, the rat models of pancreatic cancer were successfully established and 11 rats with pancreatic cancer were acquired, as well as 9 rats in the blank control group were acquired. The median expression levels of serum miRNA-21 and miRNA-155 were 2.12 (1.33–2.72) and 16.45 (7.18–25.40) in the rat pancreatic cancer group, as well as 1.00 (0.45–1.60) and 1.49 (1.25–1.97) in the blank control group. The expression levels of serum miRNA-21 and miRNA-155 were significantly higher in the rat pancreatic cancer group (Z=–2.621,P=0.009; Z=–3.609,P<0.001). For distinguishing pancreatic cancer from benign diseases, the best cutoff value of serum miRNA-21 level was 4.21 and the sensitivity and specificity were 75.0% and 61.1% respectively; the best cutoff value of serum miRNA-155 level was 4.67 and the sensitivity and specificity both were 83.3%. Conclusions The serum miRNA-21and miRNA-155 levels are elevated both in patients and rats with pancreatic cancer. Detection of serum miRNA-155 will be helpful to some extent to distinguish pancreatic cancer from benign diseases.
Objective To investigate the effect of ultra-filtration on reducing the matrix effects of the immersionof recombination human acellular dermal matrix (rhADM) on detecting residual bovine serum albumin (BSA) by ELISA.Methods Preparation of rhADM immersion: rhADM were rinsed, and then rhADM immersion were prepared. Physiologicalsal ine was used as immersion medium. Presaturation and ultra-filtration: marked the ultra-filtration tubes as PR1 (presaturation protocol 1), PR2 (presaturation protocol 2) and rhADM, respectively, added 2 mL of 1 mg/mL and 10 μg/mL BSA solution into PR1 and PR2 respectively, and added 2 mL of rhADM immersion into rhADM tubes (rhADM1 and rhADM2). The tubes were then centrifuged at 1 500 × g for 20 minutes. The above steps were repeated for 3 times. Take the inner-tube of ultrafiltration into unused centrifuge tube. Added 4 mL of 10 μg/mL BSA solution in PR1 and PR2 tubes, 4 mL of rhADM immersion in rhADM tubes, centrifuged at 1 500 × g for 20 minutes, and then the filtration was colleted. Detecting BSA concentration: the BSA concentrations of all samples were detected by using the quantitative measure of residual BSA ELISA kit. The recoveries of 10 μg/ mL BSA solution treated by presaturation protocol 1 and 2 were calculated (untreated 10 μg/mL BSA solution was as the basic sample, marked R10 and R20 respectively). The correlation coefficient between the logarithm of the filtrate dilution and the absorbance (A) value was calculated and compared with that of water exact without ultra-filtration. Results The BSA concentration of PR1 and R10 was (23.80 ± 1.58) μg/ mL and (9.04 ± 0.24) μg/mL, respectively. The BSA concentration of PR2 and R20 was (8.64 ± 0.24) μg/mL and (8.12 ± 1.01) μg/ mL, respectively. The average recovery of 10 μg/mL BSA was 263.4% ± 16.9% and 106.5% ± 3.0% when the ultra-filtration tubes were presaturaed by PR1 and PR2 (P lt; 0.01), respectively. The BSA recovery of PR2 met the detecting demand. The correlations between A value and sample dilution were increased, the correlationcoefficient was raised from — 0.727 to — 0.960 after rhADM immersion were treated by ultra-filtration. Conclusion Theresults show that the matrix effects can be reduced effectively by ultra-filtration, indicating that an acceptable recovery of BSA can be acquired when ultra-filtration tube is presaturated by sample water extract.
ObjectiveTo explore the effects of glycemia and serum calcium on occurrence and development of aortic root dilation disease. MethodsThe clinical data of patients with aortic root dilation who underwent surgical treatment in the Department of Cardiac Surgery of the First Affiliated Hospital of Xinjiang Medical University from January 2011 to October 2021 were retrospectively collected. They were divided into two groups according to whether they were accompanied by acute aortic dissection (Stanford type A), and were matched with the propensity scoring method. Logistic univariate and multivariate regression analyses were used to analyze the glycemia and the serum calcium of the patients in 24 hours at admission, and their receiver operating characteristic (ROC) curves were plotted. Results Finally 184 pairs of patients were matched, including 297 males with an average age of 48.76±9.62 years and 71 females with an average age of 49.97±10.97 years. There were statistical differences in ethnicity, history of hypertension, aortic root diameter, serum calcium and glycemia between the two groups (P<0.05). Logistic multivariate regression analyses results showed that age＜40 years (OR=4.106, P=0.010), Han nationality (OR=2.863, P<0.001), aortic root diameter＜45 mm (OR=5.063, P<0.001), hypertension (OR=2.736, P=0.001), hyperglycemia (OR=4.426, P<0.001) and hypocalcemia (OR=5.375, P<0.001) were independent risk factors for aortic root dilation disease with dissection. ROC curve analysis suggested that the area under the curve (AUC) of glycemia was 0.742 and the AUC of serum calcium was 0.737, all of which had some predictive value. Conclusion Hyperglycemia and hypocalcemia are risk factors for the development of aortic root dilation disease, and to some extent, they can be used as indicators for screening high-risk patients with aortic root dilation disease.
Objective To investigate the association of serum albumin and relevant composite indicators with malignant brain edema after acute ischemic stroke. Methods We screened patients with acute ischemic stroke admitted to the Department of Neurology, West China Hospital of Sichuan University between January and December 2022. The case group consisted of patients who developed malignant brain edema within 7 days of admission, while the control group consisted of patients who did not develop malignant brain edema within 7 days of admission. Multivariate logistic regression analysis was used to explore the association of serum albumin and relevant composite indicators with malignant brain edema after acute ischemic stroke. Results Finally, 428 patients were included, aged 70.00 (58.00, 82.00) years, with females accounting for 40.9% (n=175). The time from onset to admission was 10.00 (4.00, 24.00) hours. Forty-three patients (10.0%) developed malignant brain edema and were classified as the case group, and their onset time of malignant brain edema was 34.00 (22.50, 56.50) hours after the onset of the disease. Multivariate logistic regression analysis showed that the increase in the score of the baseline National Institutes of Health Stroke Scale scores [odds ratio (OR)=1.167], the combination of diabetes (OR=5.525), the treatment of thrombectomy (OR=23.875), and the neutrophil percentage-to-albumin ratio higher than the median (OR=3.806) were associated with the increased risk of malignant brain edema (P<0.05), and the successful reperfusion after thrombectomy (OR=0.120) was associated with the reduced risk of malignant brain edema (P<0.05). Conclusion A higher percentage of serum neutrophil percentage-to-albumin ratio within 24 hours of onset in patients with acute ischemic stroke is associated with an increased risk of malignant brain edema within 7 days of admission.
ObjectiveTo analyze the thyroid hormone levels in patients with acute type A aortic dissection (ATAAD) and assess its clinical significance.MethodsWe included 88 patients with ATAAD who underwent surgical treatment in Beijing Anzhen Hospital between January 2018 and August 2018. Meanwhile, we extracted 187 blood samples of healthy people from our laboratory (Beijing Lab for Cardiovascular Precision Medicine, Beijing, China) as control group. Examining preoperative thyroid hormone levels and perioperative serum creatine for patients and examining thyroid hormone levels for healthy people. Based on difference in thyroid hormone levels between patients and healthy people, we divide patients into abnormal thyroid hormone groups and control groups, analyzing the relationship between thyroid hormone levels and variance of postoperative serum creatinine.ResultsPatients with ATAAD have lower total triiodothyronine (TT3), thyrotropin (TSH), free triiodothyronine (FT3) and higher free thyroxine (FT4) levels than healthy people (respectively, P<0.001, P<0.001, P<0.001 and P<0.001). What’s more, patients with ATAAD who had low TT3 before operation had higher elevation of postoperative serum creatinine and rate of acute kidney injury(P=0.019).CONCLUSIONSPatients with ATAAD have different thyroid hormone levels than healthy people, preoperative TT3 is associated with elevation of postoperative serum creatinine and occurrence of acute kidney injury. Thyroid function measurement should be a routine preoperative examination in patients with ATAAD.