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.
Objective To evaluate the early diagnostic value of ischemia modified albumin (IMA) for non-ST-segment elevation acute coronary syndromes (NSTEACS). Methods The study group consisted of 177 patients with suspected NSTEACS whose blood was collected within six hours after the onset of chest pain to determine cardiac troponin I (cTnI), and IMA was determined through the albumin cobalt binding (ACB) test. After standardized diagnosis and treatment and GRACE risk score, the patients then were divided into three groups according to the final diagnosis: the NSTEMI (non-ST-segment elevation myocardial infarction) group (n=34), the UA (unstable angina pectoris) group (n=56) and the NICP (non-ischemia chest pain) group (n=87). Meanwhile, 58 people taking the routine examination in the same hospital at that time were randomly selected as the control group. With the results of IMA, ROC curve analysis was used to determine the optimal cutoff of this assay for identifying patients with NSTEACS from those with NICP. Results of IMA, ECG and cTnI were correlated with final diagnosis, and their diagnostic sensitivity and specificity were evaluated for NSTEACS. Results The IMA concentration in the serum showed no significant difference between the NSTEMI group and the UA group, whereas there were significant differences between the former two groups and the NICP group. The sensitivity and specificity at a cutoff point 67.49 U/mL were 91.1% and 86.2%, respectively when the ROC curve area was 0.950. The correlation between the IMA concentration and GRACE risk score was negative. Conclusion IMA is an early sensitive indicator for NSTEACS and a useful predictor of prognosis.
Objective To prepare the immunonanospheres［SC3Ab-HSA(5-Fu)-NS］ against human colorectal cancer and evaluate its immunoreactivity and effects on cancer. Methods SC3Ab-HSA(5-Fu)-NS was prepared by intermolecular cross-linking the monoclonal antibody SC3Ab with human serum albumin nanospheres containing 5-Fu ［HAS(5-Fu)-NS］ via new hetero-bifunctional crosslinker SPDP. Condensation test and immunoflurecence were used to evaluate the immunoreactivity, the specific binding of SC3Ab-HSA(5-Fu)-NS with colorectal cancer cell line SW1116 was observed by microscope and electron microscope. The specific cytotoxic effects on target cells were evaluated in vitro by MTT assay. SC3AbHSA(5-Fu)-NS, HSA(5-Fu)-NS and 5-Fu were injected into nude mice bearing human colorectal carcinoma, to study the inhibitory activity of SC3Ab-HSA(5-Fu)-NS in vivo. Results The immunoreactivity of SC3Ab-HSA(5-Fu)-NS was well preserved. SC3Ab-HSA(5-Fu)-NS can bind the SW1116 cells specifically. The IC50 value for SC3Ab-HSA(5-Fu)-NS on SW1116 cells was 24.6 μg/ml,which was lower than that of HSA(5-Fu)-NS(345.3 μg/ml) and 5-Fu(325.6 μg/ml). The inhibitory rate of SC3Ab-HSA(5-Fu)-NS on the growth of colorectal cancer xenografts was significantly higher than that of HSA(5-Fu)-NS or 5-Fu(P＜0.001).Conclusion SC3Ab-HSA(5-Fu)-NS has immunoreactivity and specific active targeting to the colorectal cancer cells. The anticancer ability of SC3Ab-HSA(5-Fu)-NS is significantly higher than that of HSA(5-Fu)-NS and 5-Fu.
Objective To measure the concentration of serum transthyretin (TTR) of patients with different stages of diabetic retinopathy (DR). Methods A total of 176 patients with diabetes mellitus were included in this study. There were 104 males and 72 females. The patients aged from 21 to 74 years, with the mean age of (56±11) years. The diabetes duration raged from 1 to 30 years, with the mean diabetes duration of (10±7) years. The HbA1C was 5.2%−14.1%, with the mean HbA1C of (8.6±2.0)%. According to the fundus examination, 58 patients had DR (33.0%), but the other 118 patients not (67.0%). For these DR patients, 10 patients were in stage Ⅰ (5.7%), 26 patients in stage Ⅱ (14.8%), 8 patients in stage Ⅲ (4.5%), and 14 patients in stage Ⅳ (8.0%). The concentration of serum TTR was measured by enzyme-linked immunosorbentassay kit. The differences in the concentration of serum TTR between different DR stages were compared.Bivariate analysis was used to analyze the influencing factors of TTR. Results The concentrations of serum TTR of the patients without DR or with DR of stage Ⅰ to Ⅳ were (224.96±65.47), (383.68±102.99), (247.44±63.21), (228.2±45.89), (189.34±70.12) mg/L, respectively. The difference between different DR stages was statistically significant (F=14.690,P＜0.001).Bivariate analysis showed that the concentration of TTR was correlation to DR (r=0.179,P=0.017). There was no correlation between the concentration of TTR and diabetes duration (r=−0.027,P=0.727), hypertension (r=0.018,P=0.810), hyperlipoidemia (r=0.101,P=0.182), and the use of insulin (r=−0.032,P=0.675). Conclusion The concentration of serum TTR was increased in early DR patients, and gradually decreased with the progression of DR. The concentration of TTR is correlated to DR.
Objective To investigate the risk factors of high peritoneal transport characteristics in patients with end-stage renal disease undergoing initial continuous ambulatory peritoneal dialysis. Method The clinical data of continuous ambulatory peritoneal dialysis patients who underwent initial peritoneal dialysis and catheterization in the Department of Nephrology, West China Hospital of Sichuan University from January 2011 to December 2017 and completed the peritoneal equilibration test were collected retrospectively. According to the ratio of dialysate to plasma ratio for creatinine at 4 hour [D/Pcr (4h)] in the standard peritoneal equilibration test, the patients were divided into 4 groups (low transport, low average transport, high average transport and high transport). Spearman correlation analysis was used to analyze the related factors of D/Pcr (4h). The risk factors of high peritoneal transport characteristics were analyzed by ordered multi classification logistic regression. Results A total of 647 patients were included. The average age of the patients was (45.85±14.03) years, and the average D/Pcr (4h) was 0.67±0.12. Among them, there were 89 cases (13.76%) in the high transport group, 280 cases (43.28%) in the high average transport group, 234 cases (36.17%) in the low average transport group and 44 cases (6.80%) in the low transport group. Diabetic patients with D/Pcr (4h) were higher than those without diabetes mellitus (0.72±0.12 vs. 0.66±0.12; t=−4.005, P<0.001). Correlation analysis showed that age and 24-h urine volume were positively correlated with D/Pcr (4h); serum albumin, triglyceride, potassium, calcium, magnesium, phosphorus, hemoglobin, serum uric acid and creatinine were negatively correlated with D/Pcr (4h); body surface area (BSA), high sensitivity C-reactive protein, ferritin, cholesterol, sodium, intact parathyroid hormone and estimated giomerular filtration rate had no correlation with D/Pcr (4h). Regression analysis showed that serum albumin [odds ratio (OR)=0.842, 95% confidence interval (CI) (0.809, 0.877), P<0.001], serum uric acid [OR=0.996, 95%CI (0.994, 0.998), P<0.001], magnesium [OR=0.389, 95%CI (0.156, 0.965), P=0.042], BSA [OR=3.916, 95%CI (1.121, 13.680), P=0.032] were correlated with the incidence of peritoneal high transport characteristics. Conclusion Low serum albumin, high BSA, low magnesium and low serum uric acid were independent risk factors for high transport characteristics in initial PD patients.
ObjectiveTo evaluate the prognostic value of the easy albumin-bilirubin (EZ-ALBI) score for postoperative complications and long-term prognosis of hepatocellular carcinoma (HCC) patients. MethodsThe data on consecutive 1 822 HCC patients who underwent hepatectomy were obtained and retrospectively analyzed from five medical centers, including West China Hospital, Sichuan Provincial People’s Hospital, The First People’s Hospital of Neijiang City, The Second People’s Hospital of Yibin City, and People’s Hospital of Leshan City. Non-conditional logistic and Cox proportional hazards regression were used to evaluate the aspect on the postoperative complications and long-term prognosis. ResultsThe patients in EZ-ALBI grade 2 had higher incidences of severe complication (Clavein-Dindo classification>2, P=0.001), post-hepatectomy liver failure (P=0.040), length of stay>10 d (P<0.001), perioperative transfusion (P<0.001), and 90 d mortality (P<0.001). The 1-, 3- and 5-year cumulative survival rates in EZ-ALBI grade 1 group were 85.5%, 67.0%, and 58.7% while in EZ-ALBI grade 2 group were 72.7%, 51.1%, and 39.8%. Multivariate Cox proportional hazards regression manifested that patients in EZ-ALBI grade 2 had a significantly worse overall survival [HR=1.24, 95%CI (1.04, 1.48), P=0.015]. ConclusionThe EZ-ALBI score is an easy and feasible classifying method to predict postoperative complications and survival of HCC.
ObjectiveTo investigate the predictive value of preoperative plasma fibrinogen and serum albumin score (FA score) for postoperative survival of hepatocellular carcinoma (HCC) after hepatectomy.MethodWe retrospectively analyzed the clinicopathological data and follow-up information of 275 patients with HCC who underwent hepatectomy in West China Hospital of Sichuan University from March 2009 to December 2013.ResultsThere’s no statistically significant difference in gender, ALT, total bilirubin, hepatitis B virus surface antigens, AFP, cirrhosis, macrovascular invasion, tumor differentiation, TNM stage, and postoperative adjuvant transarterial chemoembolization of HCC patients between FA score of 0 group and FA score of 1 and 2 group (P>0.05). There’s statistically significant difference in age, AST, tumor size, tumor number, microvascular invasion, and BCLC stage (P<0.05). Multivariate Cox proportional hazard regression analyses revealed that FA score (1 and 2) was an independent risk factor for HCC patients’ overall survival rate [HR=1.632, 95%CI was (1.141, 2.335), P=0.007] and early recurrence-free survival rate [HR=1.678, 95%CI was (1.083, 2.598), P=0.021], the overall survival rate and early recurrence free survival rate of HCC patients with FA score of 0 group were better than those of patients with FA score of 1 and2 group.ConclusionsThe preoperative FA score has a good prognostic value for survival of HCC patients who underwent hepatectomy. Preoperative FA score of 1 and 2 is an independent risk factor for overall survival rate and early recurrence free survival rate of HCC patients after hepatectomy.
Objective To search evidence of angiotensin-converting-enzyme inhibitors for microalbumin-uria in type 2 diabetes for guiding clinical practice. Methods We searched MEDLINE （ 1970 -Jun. 2005 ） to identify randomized controlled trials （RCT）of the effect on angiotensin-converting-enzyme inhibitors to prevent microalbuminuria in type 2 diabetes. Results One RCT （n =1 204）was identified. The result showed that angiotensin-converting-enzyme inhibitors were significantly more effective in prevention of microalbuminuria than other medicines in type 2 diabetes. However, angiotensin-converting-enzyme inhibitors may increase the risk of cardiac mortality. We explained the evidence to patients and they were satisfied with our explanation. Conclusions Angiotensin-converting-enzyme inhibitors can decrease the incidence of microalbuminuria in patients with type 2 diabetes and hypertension.
ObjectiveTo observe and evaluate the predictive value of serum cystatin C (Cys-C) on the risk of sight-threatening diabetic retinopathy (STDR). MethodsA non-randomized controlled cross-sectional clinical study. Ninety-two patients with type 2 diabetes mellitus (T2DM) who were admitted to Department of Ophthalmology of Beijing Tsinghua Changgung Hospital from January 2022 to October 2022 were included in the study. Among them, 50 were male, 42 cases were female, with the mean age of (58.24±12.49) years. The mean duration of T2DM was (13.18±8.35) years, of which 38 cases had a duration of ≥10 years. Twenty-nine cases complicated with hypertension, of which 16 cases had a duration of ≥10 years. Seventeen cases complicated with chronic kidney disease stage 2 and 23 cases were treated with lipid-lowering drugs. Hemoglobin Alc, serum Cys-C, serum lipids and renal function were tested, and urinary microalbumin/creatinine ratio (ACR) was calculated. According to the 2003 American Academy of Ophthalmology "Clinical Guidelines for Diabetic Retinopathy (DR)" and international clinical DR severity grading standards, the patients were divided into STDR and non-STDR groups, with 44 and 48 cases in each group, respectively. STDR was defined as severe non-proliferative DR, proliferative DR, and macular edema. Logistic regression was used to analyze the independent risk factors of STDR in T2DM patients. Receiver operating characteristic curve (ROC curve) was used to calculate and analyze the area under ROC curve (AUC) and the predictive value of serum Cys-C and ACR in predicting STDR in T2DM patients. ResultsSerum Cys-C levels in STDR and non-STDR groups were 1.10 (0.94, 1.28) and 0.91 (0.83, 1.02) mg/L, respectively, with ACR of 4.29 (1.05, 21.89) and 1.39 (0.77, 3.80) mg/mmol, respectively. Compared with non-STDR group, serum Cys-C and ACR in STDR group were higher, and the difference was statistically significant (Z=－3.984, －3.280; P＜0.05). Multivariate logistic regression analysis showed that serum Cys-C was an independent risk factor for STDR (odds ratio=1.337, 95% confidence interval 1.145-2.090, P=0.033), and the risk of STDR increased by 33.7% for every 0.1 mg/L increase in serum Cys-C. ROC analysis results showed that serum Cys-C＞1.065 mg/L combined with ACR＞5.84 mg/mmol predicted the AUC of STDR in T2DM patients was 0.661, with the specificity of 95.8%. ConclusionsThe high serum Cys-C level is an independent risk factor for STDR in T2DM patients. Serum Cys-C has high predictive value for the occurrence of STDR.
ObjectiveTo investigate the effect of maresin-1 (MaR1) on lung inflammation and MAPK signaling pathway in asthmatic mice.MethodsTwenty-four female BALB/c mice were randomly divided into normal group, asthma model group, MaR1 group and dexamethasone group. The asthma model was successfully established by using ovalbumin (OVA) combined with aluminum hydroxide, and then MaR1 and dexamethasone were respectively given to asthmatic mice. Serum, bronchoalveolar lavage fluid (BALF) and lung tissue were collected for further analysis. Pathological changes of lung tissue in mice were detected by hematoxylin-eosin and periodic acid-Schiff. Proportion of inflammatory cells in BALF classified by Swiss-Giemsa staining. Th2-related inflammatory cytokines, interleukin (IL)-4, IL-5, IL-13 and IgE in serum and BALF were detected by enzyme linked immunosorbent assay. The protein concentration of p-p38 and p-JNK in lung tissues were detected by Western blot.ResultsCompared with the normal group, the asthma model group had increased both airway inflammation and the number of goblet cells significantly (P<0.05). The number of various inflammatory cells in BALF had also increased significantly (P<0.05). The levels of IL-4, IL-5 and IL-13 in BALF and IgE and OVA-specific-IgE in serum were significantly increased (P<0.05). The protein contents of p-p38 and p-JNK in lung tissues were significantly increased (P<0.05). Compared with the asthma model group, both MaR1 and dexamethasone group had reduced inflammation and mucus secretion in lung tissue, number of inflammatory cells in BALF (P<0.05), levels of related inflammatory cytokines in BALF and IgE in serum (P<0.05), and expression of p-p38 and p-JNK proteins in lung tissue (P<0.05).ConclusionsMaR1 can inhibit the production and release of both Th2-related inflammatory cytokines and IgE, effectively reduce the inflammatory response and mucus production in lung tissues of asthmatic mice, with similar effect to dexamethasone. The mechanism may be related to the down-regulation of MAPKs signaling pathway.