Objective To establish a purified model of rat retinal ganglion cells （RGCs） cultured by serum-free medium，and provide a good cell model to investigate the damage of RGCs in glaucoma，retinal ischemia，and degenerative retinopathy. Methods Two monoclonal antibodies，anti-rat SIRP（OX-41）against rat macrophage and antibody against rat Thy-1（OX-7），were used to purify and characterize RGCs from 1-3-day old Sprague-Dawley（SD）rats by means of two-step filtration.Purified RGCs were cultured in serum-free neurobasal medium containing B27 and ciliary neurotrophic factor（CNTF） meeting the neuronal cellrsquo;s special requirements.Photomicrographs illustration，immunfluorescence staining of Thy-1，calcein-acetoxymethyl ester（calcein-AM）fluorescence images were used to observe and identify cultured retinal cells and purified RGCs. Results Among the primary cultured rat retinal cells，91% were retinal neurons.Protuberances of RGCs were seen after cultured for 24 hours.At the4th to 8th day，many cells had uniform configuration，large body，and long protuberances. At the 14th day，over 60% cells maintained viability.Immunoflurescence staining of Thy-1 showed the purity of RGCs was about 90%. The results of calcein-AM staining，which stained the living cells only，showed large cell body of RGCs and most of RGCs had a protuberance whose length was twice longer than the diameter of the cells. Conclusion RGCs cultured by serum-free medium has uniform size，good configuration，and high purity，which is adapt to the research of damage of RGCs caused by various factors and to evaluate the protective effects of neuroprotective agents. (Chin J Ocul Fundus Dis, 2006, 22: 200-203)
Objective To investigate the possibility of culturing human oral keratinocyte using autologous serum in order to provide theoretical and technical foundation for clinical application of tissue engineering oral mucosa epithelium.Methods The human oral keratinocytes were cultured by the medium containing different concentrations of autologous serum(10%,20%,30%)and fetalbovine serum (10%), respectively. The growth conditions for the cell and the mucosa epithelium in the groups were observed, the cell growth curves were drawn, and the population doubling time (PDT) was counted. Results The results showed that the human oral keratinocyte could proliferate well in the medium containing autologous serum or fetal bovine serum. The differences in the 24hour clone rate and PDT were not significant. Both the area and the thickness of the cultured oral epithelium increased with the increase of the autologous serum concentration, and the difference between autologous serum and fetal bovine serum was significant, especially with the medium containing 20% autologous serum( P＜0.05) . The human nature of the cultured epithelium was demonstrated by the immunofluorescent mouse anti-HLA antigen. Conclusion The autologous serum can replace the fetal bovine serum to culture the oral keratinocyte well, and the cultured oral mucosa epithelium can be better differentiated in the autologous serum than in the fetal bovine serum.
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.
Delirium is an acute cognitive disorder caused by a variety of factors which lead to cerebral cortical dysfunction. At present the studies on the pathophysiology of delirium is still very few. But studies on serum biomarker of delirium can help to elucidate the pathophysiological mechanism of delirium, and the studies are significant for delirium diagnosis, severity classification and prediction of long-term outcome. This review examines three major groups of delirium related serum biomarkers: ① risk markers: those that are present or elevated prior to disease onset, including serum chemistries, genetic markers and so on; ② disease markers: those markers elevate with delirium onset and fall when delirium recovery, including acetylcholine and serum anticholinergic activity, serotonin, serum amino acids, and melatonin, interleukin, C-reactive protein; and ③ end products: those that rise in proportion to the consequences of disease, including S-100ß and neuron specific enolase. The three markers mentioned above are helpful to further investigate the mechanism of delirium.
Hypernatremia is one of the commonly syndromes in critically ill patients. Severe hypernatremia has a low incidence (0.6%–1.0%) but with a very high mortality (58%–87%). Conventional treatments include the limitation of sodium intake and the supplement of sodium free liquid according to the assessed water lost. The reduction rates of conventional treatments are commonly not effective enough to decrease the serum sodium concentration in severe euvolemic or hypervolemic hypernatremia patients. Continuous renal replacement therapy (CRRT) has been reported to be effective on the reduction of sodium level in severe hypernatremia patients. However, the evidences on the use of CRRT for hypernatremia are limited. Our present review summarizes the current evidences on the prevalence of hypernatremia, the outcome of hypernatremia patients, the conventional treatment of hypernatremia, and the advantages and indications of CRRT for the management of hypernatremia. Additionally, we introduce our experiences on the management of hypernatremia using CRRT as well.
ObjectiveTo investigate the prognostic value of preoperative serum albumin-to-globulin ratio (AGR) and neutrophil-lymphocyte ratio (NLR) in the overall survival (OS) of patients with esophageal squamous cell carcinoma (ESCC), and to establish an individualized nomogram model and evaluate its efficacy, in order to provide a possible evaluation basis for the clinical treatment and postoperative follow-up of ESCC patients. MethodsAGR, NLR, clinicopathological and follow-up data of ESCC patients diagnosed via pathology in the Department of Thoracic Surgery, The First Affiliated Hospital of Xinjiang Medical University from 2010 to 2017 were collected. The correlation between NLR/AGR and clinicopathological data were analyzed. Kaplan-Meier analysis and log-rank test were used for survival analysis. The optimal cut-off values of AGR and NLR were determined by X-tile software, and the patients were accordingly divided into a high-level group and a low-level group. At the same time, univariate and multivariate Cox regression analyses were used to identify independent risk factors affecting OS in the ESCC patients, and a nomogram prediction model was constructed and internally verified. The diagnostic efficacy of the model was evaluated by receiver operating characteristic (ROC) curve and calibration curve, and the clinical application value was evaluated by decision curve analysis. ResultsA total of 150 patients were included in this study, including 105 males and 45 females with a mean age of 62.3±9.3 years, and the follow-up time was 1-5 years. The 5-year OS rate of patients in the high-level AGR group was significantly higher than that in the low-level group (χ2=6.339, P=0.012), and the median OS of the two groups was 25 months and 12.5 months, respectively. The 5-year OS rate of patients in the high-level NLR group was significantly lower than that in the low-level NLR group (χ2=5.603, P=0.018), and the median OS of the two groups was 18 months and 39 months, respectively. Multivariate Cox analysis showed that AGR, NLR, T stage, lymph node metastasis, N stage, and differentiation were independent risk factors for the OS of ESCC patients. The C-index of the nomogram model was 0.689 [95%CI (0.640, 0.740)] after internal validation. The area under the ROC curve of predicting 1-, 3-, and 5-year OS rate was 0.773, 0.724 and 0.725, respectively. At the same time, the calibration curve and the decision curve suggest that the model had certain efficacy in predicting survival and prognosis. ConclusionPreoperative AGR and NLR are independent risk factors for ESCC patients. High level of AGR and low level of NLR may be associated with longer OS in the patients; the nomogram model based on AGR, NLR and clinicopathological features may be used as a method to predict the survival and prognosis of ESCC patients, which is expected to provide a reference for the development of personalized treatment for patients.
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.
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.
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 investigate the effect of asymptomatic hyperuricemia on the effectiveness of arthroscopic rotator cuff repair.MethodsThe clinical data of 80 patients who underwent arthroscopic rotator cuff repair and met the selection criteria between March 2018 and December 2019 were retrospectively analyzed. According to the serum uric acid level, the patients were divided into hyperuric acid group (46 cases, the serum uric acid level was more than 417 μmol/L in males and was more than 357 μmol/L in females) and normal group (34 cases, serum uric acid level was lower than the above standard). There was no significant difference in gender, age, side, body mass index, blood glucose level, total cholesterol level, rotator cuff tear size, and preoperative shoulder motion, visual analogue scale (VAS) score, University of California-Los Angeles (UCLA) score, Constant score, American Shoulder and Elbow Surgeons (ASES) score, and other general data between the two groups (P>0.05). The range of motion of abduction, forward flexion, and external rotation at 90° abduction were recorded and compared between the two groups before operation and at last follow-up; the improvement of shoulder pain was evaluated by VAS score; the improvement of shoulder function was evaluated by UCLA score, Constant score, and ASES score; and the shoulder joint MRI grade was evaluated according to Sugaya evaluation criteria.ResultsAll patients were followed up 9-16 months, with an average of 11.9 months; there was no significant difference in the follow-up time between the two groups (t=0.968, P=0.336). There were 2 cases of retear in the hyperuric acid group (including 1 case of severe tear) and 1 case of light retear in the normal group. The remaining patients in the two groups had no early-related complications. At last follow-up, the range of motion of the shoulder joints (abduction, forward flexion, external rotation at 90° abduction), VAS score, UCLA score, Constant score, and ASES score of the two groups were significantly improved when compared with preoperative ones (P<0.05); the above indicators in the normal group were significantly better than those in the hyperuric acid group (P<0.05). The MRI grade of the shoulder joint in the normal group was significantly better than that in the hyperuric acid group (Z=–2.000, P=0.045).ConclusionCompared with patients with normal serum uric acid level, asymptomatic hyperuricemia can lead to worse recovery after arthroscopic rotator cuff repair in patients with rotator cuff tears.