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AIM: To explore the correlation by analyzing and comparing the expression of inflammatory factors in peripheral blood of human leukocyte antigen-B27(HLA-B27)positive and negative uveitis patients.METHODS: All 76 patients first diagnosed with uveitis in our hospital from January 2020 to April 2022 were screened in this retrospective study. Nucleated cells were isolated from human venous blood, and HLA-B27 was detected by flow cytometry(direct immunofluorescence), the patients were divided into the HLA-B27-positive group(≥90%)in 35 cases and HLA-B27-negative group(≤5%)in 41 cases. The whole blood RNA was extracted. The mRNA expression of interleukin-1β(IL-1β), interleukin-6(IL-6), interleukin-10(IL-10), protease activated receptor 2(PAR2), tumor necrosis factor-α(TNF-α), interleukin enhancer-binding factor -3(ILF3)were detected and compared by RT-qPCR.RESULTS: The IL-1β, IL-10, PAR2 and TNF-α mRNA were observed no difference between the two groups of patients(all P>0.05). The IL-6 mRNA in the patients of HLA-B27-positive group was higher than in the HLA-B27-negative group, the ILF3 mRNA was lower than that in the negative group(all P<0.05).CONCLUSION: The expression level of IL-6 in peripheral blood was significantly increased and ILF3 was decreased in HLA-B27-positive group, which can be used as auxiliary indicators for diagnosis and treatment of HLA-B27-positive uveitis.
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<p><b>Background</b>Measuring total serum calcium is important for the diagnosis of diseases. Currently, results from commercial kits for calcium measurement are variable. Generally, the performance of serum calcium measurements is monitored by external quality assessment (EQA) or proficiency testing schemes. However, the commutability of the EQA samples and calibrators is often unknown, which limits the effectiveness of EQA schemes. The aim of this study was to evaluate the bias of serum calcium measurements and the commutability of processed materials.</p><p><b>Methods</b>Inductively coupled plasma mass spectrometry was applied as a comparative method, and 14 routine methods were chosen as test methods. Forty-eight serum samples from individual patients and 25 processed materials were quantified. A scatter plot was generated from patient samples, and 95% prediction intervals were calculated to evaluate the commutability of the processed materials and measurement bias at three concentration levels was used to determine the accuracy of routine assays.</p><p><b>Results</b>All assays showed high precision (total coefficient of variation [CV] <2.26%) and correlation coefficients (r > 0.99). For all assays, the mean bias for the 48 patient samples ranged from -0.13 mmol/L to 0.00 mmol/L (-5.61-0.01%), and the ranges for the three concentrations were -0.10-0.04 mmol/L (-5.71-2.35%), -0.14--0.01 mmol/L (-5.80--0.30%), and -0.19-0.04 mmol/L (-6.24-1.22%). The EQA samples, calibrators, and animal sera exhibited matrix effects in some assays; human serum pools were commutable in all assays; certificate reference materials were commutable in most assays, and only GBW09152 exhibited a matrix effect in one assay; and aqueous reference materials exhibited matrix effects in most assays.</p><p><b>Conclusions</b>Biases for most assays were within the acceptable range, although the accuracy of some assays needs improvement. Human serum pools prepared from patient samples were commutable, and the other tested materials exhibited a matrix effect.</p>
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Abstract?AIM: To investigate the influence of propylene glycol mannite sulfate ( PGMS ) on the expression of tumor necrosis factor -α( TNF-α) and interleukin-1β( IL-1β) , in diabetic retinopathy by a rat model, to study the mechanism of PGMS against diabetic retinopathy, and provide a reliable theoretical and experimental evidence for the PGMS to be applied to clinical prevention and treatment of diabetic retinopathy.?METHODS: Male Wistar rats were randomized into 4 groups, normal control group, diabetic control group and PGMS in group, the PGMS in groups included the doses of 50mg/kg and 100mg/kg. 1% streptozotocin ( STZ) of 60 mg/kg was intraperitoneally injected in rats to establish the diabetic models. The PGMS with the doses of 50mg/kg and 100mg/kg were used to gavage in different groups of models for 12wk.Twelve weeks later, the animals were sacrificed and retinas were isolated. The aqueous humors and serums were taken, expressions of TNF-αand IL-1βprotein in retinas, aqueous humors and serums were detected by enzyme-linked immunosorbent assay ( ELISA) , respectively.The location and the expression of TNF-αand IL-1βprotein in retina tissue was detected by immunohistochemistry.?RESULTS: Twelve weeks after the use of PGMS, the level of blood glucose was not changed.ELISA showed that the expression of TNF-αand IL-1βprotein in serum and retina was significantly increased in diabetic control group than in normal control group(P<0.05), but in the groups which PGMS was given reduced, lower than those in diabetes mellitus( DM) group, especially as the concentration of PGMS increased ( P<0.05 ).But the levels of aqueous humor's TNF-αand IL-1βproteins in PGMS group were not reduced.Immunohistochemistry showed that the TNF -α protein was almost not expressed in normal control group. But the TNF-αprotein was highly expressed in diabetic control group. The expression mainly located in the ganglion cell layer, the inner plexiform layer, outer plexiform layer and pigment epithelium. The TNF-αprotein was weakly expressed at the group of 50mg/kg PGMS, the TNF-αprotein was almost not expressed at the group of 100mg/kg PGMS.When the normal control group was detected, the IL-1βprotein was weakly expressed in the outer plexiform layer.But the IL-1βprotein was also highly expressed in diabetic control group.The expression mainly located in the inner plexiform layer, outer plexiform layer and pigment epithelium. The IL -1βprotein was weakly expressed at the group of 50mg/kg and 100mg/kg PGMS.?CONCLUSION:PGMS can treat the diabetic retinopathy by downregulating the expressions of TNF-αand IL-1βin early diabetic retinopathy.PGMS maybe have a good control effect on early diabetic retinopathy.
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AIM: To explore the relationship between the expression of caspase-2 and caspase-3 and the apoptosis in retinal ischemia/reperfusion (I/R) injury of rats, as well as the therapeutic effects of brain derived neurotrophic factor (BDNF)on the ischemic and reperfused retina.METHODS: This experiment was conducted at the laboratory of Affiliated Hospital of Qingdao University Medical College from February 2007 to July 2007. The models of retinal ischemia/reperfusion injury were made by transiently elevating intraocular pressure. A total of 28 rats were divided into Normal and Operative Groups. Operative group was divided into six subgroups. In each subgroup there were four rats. The left eyes of rats were used for I/R and the right eyes were used for intravitreal injection of brain-derived neurotrophic factor (BDNF) as treatment group. After reperfusion we divided our subgroups according to the reperfusion time as 1, 6, 12, 24, 48, 72 hours. The retinal ganglion cell number was counted by using optic microscope(BX-51,Olympus). Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling (TUNEL) method, and the expression of caspase-2,caspase-3 was studied by enzyme linked immunosorbent assay (ELISA) and strept avidin-biotin complex (SABC)immunohistochemistry.RESULTS: No positive apoptotic cells were observed in the normal rats' retinae, but there were a significant number of positive apoptosis cells in 6-24 hours after transient ischemia followed by a decrease at 48 hours. The number of apoptotic cells reached a maximum at 24 hours after ischemia .The expression of caspase-2 gradually increased as early as at 6 hours, reached a peak at 24 hours, then decreased between 48 and 72 hours. Similarly, caspase-3 has the same rule with caspsae-2 in the time courses of expression in retinal tissues.BDNF administered before reperfusion inhibited the expression of apoptosis and ameliorated the retinal tissue damage. It also decreased caspase-2 and caspase-3 expression in ischemic/reperfused retina.CONCLUSION: Retinal ischemia-reperfusion can induce apoptosis of cells in the retina. BDNF rescues retinal ganglion cells (RGCs) from retinal ischemia/reperfusion injury through down-regulation of cell apoptosis and caspase-2 and caspase-3 expression. BDNF have a neuroprotective effect on retina.
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Objective To develop a method for the determination of total cholesterol in serum by isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS).Methods Serum samples were supplemented by addition of [3,4-~(13)C_2]-cholesterol,hydrolyzed with alcoholic sodium hydroxide and oxidized into cholest-4-ene-3,6-dione by chromic acid.The oxidation products were analyzed by LC/MS/MS using atmospheric pressure chemical ionization (APCI) source and detection modes of multiple reaction monitoring (MRM) and single ion recording (SIR).Signals (peak areas) of the internal standard were corrected for the contributions of cholesterol and the signal ratios of cholesterol to internal standard for the calibrations were linearly regressed against cholesterol concentrations.The resulted regression equation was used for the calculation of serum cholesterol concentrations.Results The correlation coefficients between the peak area ratios and cholesterol concentrations were 0.999 9 and higher.Under MRM mode,the average within-run CV of the results obtained on 3 serum samples was 0.95% (ranged from 0.92% to 0.99%) and the total CVwas 0.86% (0.82% to 0.89%),and under SIR mode,the within-run CV was 0.64% (from 0.54% to 0.77%) and the total CVwas O.69% (0.62% to 0.81%),respectively. Results on certified reference materials (SRM 1951 a Level Ⅰ and Level Ⅱ;GBW 09145 and GBW 09147) showed an average bias of 0.23% (0.14% to 1.00%) under MRM mode,and 0.24% (0.07% to 1.27%) under SIR mode.Conclusions An ID-LC/MS/MS method for serum cholesterol has been developed.It is specific and precise and may be used as a candidate reference method.