Homologous Assay Development for Detection of Dexamethasone using HRP as Label Enzyme

Background: Dexamethasone is a synthetic corticosteroid similar to cortisol produced naturally by the adrenal glands. As an anti- inflammatory and immunosuppressive agent, it is used in many diseases such as rheumatoid arthritis and allergic anaphylactic shock, and its suppression test to diagnose Cushing's syndrome. Its further use includes its administration before antibiotics in bacterial meningitis, antitumor treatment, for treatment of glucocorticoid resistance, Addison抯 disease, and congenital adrenal hyperplasia. The drug is abused by using it in animal husbandry as a growth promoter and in horse sports to enhance their performance. Methods: In this study, the development of homologous ELISA using Dexamethasone-21-hemisuccinate (DEX-21-HS)-Bovine serum albumin antiserum and Dexamethasone-21-hemisuccinate (DEX-21-HS)-Horseradish peroxidase enzyme conjugate has been done. The n-hydroxysuccinimide ester method was used to prepare the immunogen and enzyme conjugate. Results: The sensitivity 0.25 ng/mL, affinity 2.8x10-8 L/mol and ED50 4.98 ng/mL of the assay were found. The cross-reactivity of the assay was checked and found with three steroids (Corticosterone- 1.13%, Progesterone- 2.25% and Prednisolone- 6.3%) out of 48 structurally related steroids. Then, analytical variables of the developed assay were studied, such as recovery (98.55% to 105.08%), precision (Inter and Intra- assay coefficient of variation <9.28%), correlation (R2= 0.98) by utilizing a commercially available Dexamethasone kit for comparison. Conclusion: This study concluded that low-cost indigenous ELISA for Dexamethasone had been developed, which can give results within 75-80 minutes.

Impact of antioxidant therapy on antioxidant capacity in hypertensive patients

Background: Hypertension is consistently related to the development of ischemic heart disease, heart failure, stroke, and chronic kidney disease. Oxidative stress has been associated with mechanisms of hypertension which could be nullified by antioxidants such as Vitamin C and Vitamin E. Aim and Objectives: The objectives of the study are as follows: (i) To estimate the impact of antioxidant therapy on antioxidant capacity in hypertensive patients; (ii) to measure serum levels of glutathione, glutathione peroxidase (GPx), glutathione reductase (GR), and superoxide dismutase (SOD) in hypertensive patients before and after giving them antioxidant therapy for 45 days. Materials and Methods: Thirty randomly selected hypertensive patients were given Supradyn tablet once a day for 45 days. Ferric reducing ability of plasma (FRAP), SOD, GR, GPx, and reduced Glutathione assays were measured before and after the intervention therapy. Results: Total antioxidant capacity as measured by serum FRAP in hypertensive patients before and after the therapy was increased significantly from 578.8 ± 60.85 to 592.1 ± 59.66 (?mol/L), respectively. The levels of SOD, GPx, GR, and Glutathione in hypertensive patients before giving antioxidant therapy were 1.6 ± 0.49 U/ml, 184.6 ± 17.1 ?mol/L/min, 8.96 ± 1.15 ?mol/L/min, and 8.03 ± 0.96 ?mol/g of Hb, respectively. The same after giving them antioxidant therapy were 1.7 ± 0.46 U/ml, 182.4 ± 15.98 ?mol/L/min, 8.83 ± 1.11 ?mol/L/min, and 7.83 ± 0.94 ?mol/g of Hb, respectively. The levels of GPx, GR, and Glutathione were significantly decreased after giving antioxidant therapy for 45 days while SOD level did not change significantly. Conclusion: Antioxidant therapies for 45 days led to a significant increase in total antioxidant capacity as shown by plasma FRAP levels and a significant decrease in serum levels of enzymatic antioxidants such as GPx, GR and Glutathione in hypertensive patients. However, serum levels of SOD did not show a significant change.

Influence of glutathione-related genetic variants on the oxidative stress profile of Mexican patients with psychotic disorders

Objective: The clinical trajectories of patients with psychotic disorders have divergent outcomes, which may result in part from glutathione (GSH)-related high-risk genotypes. We aimed to determine pharmacokinetics of clozapine, GSH levels, GSH peroxidase (GPx) activity, gene variants involved in the synthesis and metabolism of GSH, and their association with psychotic disorders in Mexican patients on clozapine monotherapy and controls. Methods: The sample included 75 patients with psychotic disorders on clozapine therapy and 40 paired healthy controls. Plasma clozapine/N-desmethylclozapine, GSH concentrations, and GPx activity were determined, along with genotyping of GCLC and GSTP1 variants and copy number variations of GSTP1, GSTT1, and GSTM1. Clinical, molecular and biochemical data were analyzed with a logistic regression model. Results: GSH levels were significantly reduced and, conversely, GPx activity was higher among patients than controls. GCLC_GAG-7/9 genotype (OR = 4.3, 95%CI = 1.40-14.31, p = 0.019) and hetero-/homozygous genotypes of GCLC_rs761142 (OR = 6.09, 95%CI = 1.93-22.59, p = 0.003) were found to be risk factors for psychosis. The genetic variants were not related to clozapine/N-desmethylclozapine levels or metabolic ratio. Conclusions: GCLC variants were associated with the oxidative stress profile of patients with psychotic disorders, raising opportunities for intervention to improve their antioxidant defenses. Further studies with larger samples should explore this proposal.

Retinal photoreceptor cell damage caused by diabetes through down-regulation of glutathione peroxidase 4 and its mechanism / 中华实验眼科杂志

Objective:To investigate the changes of glutathione peroxidase 4 (GPX4) in retinal photoreceptor cells, and the related mechanism correlated with retinal photoreceptor cell damage.Methods:The posterior segment tissues of 8 age-matched male donors were collected from the Body (Organ) Donation Register and Corneal Receiving Station of Tongji Hospital of Wuhan Red Cross from 2018 to 2021, including 4 non-diabetic donors and 4 diabetic donors.The tissues were divided into diabetes group and control group according to their donors.A total of 14 healthy SPF 8-week-old male C57BL/6 mice were selected and randomly divided into diabetes group and control group by the random number method, with 7 mice in each group.The mice in diabetes group were intraperitoneally injected with streptozotocin at a dose of 50 mg/kg for 5 days, and no intervention was given to mice in control group.Mouse photoreceptor cells 661W were divided into advanced glycation end products (AGEs) group and control group.AGEs group was treated with 100 μg/ml AGEs for 24 hours to simulate diabetic injury, and no intervention was given to control group.The outer segment morphology of retinal photoreceptors in human and mouse retinas was observed by hematoxylin-eosin staining.The expressions of glial fibrillary acidic protein (GFAP), rhodopsin and GPX4 in human and mouse retinas were detected by immunofluorescence staining.The expressions of GFAP, rhodopsin and GPX4 in mouse retina and the expression of GPX4 in 661W cells were determined by Western blot.The activity of 661W cells was detected by cell counting kit-8 (CCK8) method.The concentration of malondialdehyde (MDA) in mouse retina and cells was detected by TBA method.The activity of superoxide dismutase (SOD) in mouse retina and cells was detected by hydroxylamine assay.The use of human tissues was approved by the Ethics Committee of Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology (No.TJ-C20230301). The animal experiments were conducted with reference to the Standards Guide for the Care and Use of Laboratory Animals of the National Institutes of Health, and the study protocol was approved by the Experimental Animal Ethics Committee of Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology (No.TJH-2016001).Results:Hematoxylin-eosin staining showed that retinal photoreceptor outer segments were deformed or broken in diabetic donors and diabetic mice compared with control groups.GFAP fluorescent signal mainly appeared in the inner retina of human and mice, and the stained cells were spindle or polygonal, which was consistent with the shape of glial cells.The retinal GFAP fluorescent signal of diabetic tissue and mouse groups was stronger than that of respective control groups.Rhodopsin was only expressed in the outer segment layer of photoreceptors with clear boundaries, and GPX4 was expressed in the whole retina with strong signal in the outer segment layer of photoreceptors.The fluorescent signals of rhodopsin and GPX4 in diabetic tissue and mouse groups were weaker than those in respective control groups.The relative expressions of GFAP were significantly higher and the relative expressions of rhodopsin and GPX4 were significantly lower in diabetic tissue and mouse groups than in respective control groups (all at P<0.05). The cell viability of AGEs group was significantly lower than that of control group ( t=13.490, P<0.001). The relative expression of GPX4 protein in AGEs group was 0.42±0.12, which was significantly lower than 1.00±0.04 in control group ( t=9.041, P<0.001). MDA concentration was higher and SOD activity was lower in retinal tissue of diabetic mice and AGEs group than those in respective control groups, and the differences were statistically significant (all at P<0.05). Conclusions:Diabetes can reduce the GPX4 level in retinal photoreceptor cells and cause the imbalance of oxidation-antioxidant system, which may be the mechanism of the damage to retinal photoreceptor cells caused by diabetes.

The effect of nuclear factor erythroid 2 related factor 2-induced inhibition of ferroptosis on hyperoxia lung injury / 中华实用儿科临床杂志

Objective:To observe the expression changes of nuclear factor erythroid 2 related factor 2 (Nrf2) and glutathione peroxidase (GPX4) in human pulmonary microvascular endothelial cells (HPMEC) under different experimental conditions, and to explore the role of Nrf2 in inhibiting ferroptosis in the process of alleviating hyperoxic lung injury(HLI).Methods:Hyperoxic model was established by hyperoxia exposure.HPMEC were treated with blank control (control group), oxygen exposure at the concentration of 950 mL/L (hyperoxia group), oxygen exposure at the concentration of 950 mL/L+ 10 μmol/L Ferrostatin (ferroptosis inhibitor group) and oxygen exposure at the concentration of 950 mL/L + 10 μmol/L ML385 (Nrf2 inhibitor group). Cell viability at 24 h and 48 h was tested by the Cell Counting Kit-8 assay, and reactive oxygen species (ROS) levels were detected by a commercial ROS kit.The mRNA and protein levels of Nrf2 and GPX4 were detected by real-time quantitative polymerase chain reaction and Western blot, respectively.Differences were analyzed using the Student′s t-test for a two-group comparison or one-way ANOVA test among groups. Results:(1)Compared with the control group, significantly decreased viability and increased ROS levels were detected in hyperoxia group.Meanwhile, the mRNA (24 h: 0.750±0.010 vs.1.010±0.160, 48 h: 0.690±0.050 vs.1.000±0.070) and protein levels of GPX4 (24 h: 0.160±0.010 vs.0.290±0.010, 48 h: 0.190±0.010 vs.0.250±0.010) at 24 h and 48 h were significantly downregulated, while the mRNA (24 h: 1.740±0.050 vs.1.000±0.050, 48 h: 2.130±0.020 vs.1.000±0.030) and protein levels of Nrf2 (24 h: 0.840±0.010 vs.0.480±0.010, 48 h: 0.840±0.010 vs.0.550±0.030) at 24 h and 48 h were significantly upregulated in hyperoxia group than those of control group (all P<0.05). (2)Compared with the hyperoxia group, significantly increased viability and decreased ROS levels were detected in ferroptosis inhibitor group.Meanwhile, the mRNA (24 h: 1.520±0.110, 48 h: 1.880±0.050) and protein levels of GPX4 (24 h: 0.290±0.010, 48 h: 0.250±0.004) at 24 h and 48 h were significantly upregulated, while the mRNA (24 h: 0.780±0.040, 48 h: 0.760±0.030) and protein levels of Nrf2 (24 h: 0.480±0.010, 48 h: 0.540±0.020) at 24 h and 48 h were significantly downregulated in ferroptosis inhibitor group than those of hyperoxia group (all P<0.05). (3)Compared with the hyperoxia group, significantly decreased viability and increased ROS levels were detected in Nrf2 inhibitor group.Meanwhile, the mRNA (24 h: 0.600±0.030, 48 h: 0.590±0.003) and protein levels of GPX4 (24 h: 0.150±0.001, 48 h: 0.180±0.001) at 24 h and 48 h were significantly downregulated, while the mRNA level of Nrf2 was significantly upregulated at 24 h (3.360±0.130), but downregulated at 48 h (1.430±0.130) (all P<0.05). No significant difference was detected in the protein level of Nrf2 at 24 h and 48 h between hyperoxia group and Nrf2 inhibitor group ( P>0.05). Conclusions:Ferroptosis is involved in the development of HLI, and Nrf2 is able to alleviate hyperoxic lung injury by inhibiting ferroptosis.Therefore, inhibition of ferroptosis by Nrf2 may provide a new therapeutic target for HLI.

Correlation analysis of TPO antibody and thyroglobulin antibody with clinicopathological features of PTC / 中华内分泌外科杂志

Objective:To explore and analyze the correlation between thyroid peroxidase (TPO) and thyroglobulin (Tg) in patients with papillary thyroid carcinoma (papillary thyroid carcinoma, PTC) and to provide a more reasonable plan for the clinical diagnosis and treatment of patients.Methods:A retrospective analysis was made of 142 PTC patients who underwent surgical resection from Jun. 2019 to Jun. 2022 in the Oncology Department of Wenzhou Hospital of Traditional Chinese Medicine. 115 patients were selected, including 25 males (21.74%), and 90 females (78.26%), and the average age was (43.48±9.74) years old. The medical records, pathology reports, and demographic characteristics and pathological characteristics were collected. Immunohistochemical staining was used to detect the expression of TPO and Tg in PTC tissues, which were divided into positive and negative groups. Multifactorial Logistic regression analysis was used to analyze its relationship with clinicopathological characteristics and prognosis of patients.Results:The negative rate of TPO was 95.45% (105 cases). Univariate analysis showed that the tumor diameter ( t=5.746), lymph node metastasis, and the proportion of PT1 patients were significantly different between the two groups ( P<0.05), the TPO negative group was significantly higher than the positive group. Multivariate logistic regression analysis found that tumor diameter, lymph node metastasis, and proportion of PT1 patients were independent factors (95% CI=2.367-5.365, 1.101-2.738, 1.103-2.589, P<0.05). The positive rate of Tg was 77.41% (89 cases). Univariate analysis showed the proportion of people with BMI ≥ 25 ( χ2=11.180), tumor diameter ( t=2.117), and intracapsular invasion ( χ2=8.354), extrathyroidal invasion, lymph node metastasis ( χ2=27.740), and proportion of PT1 patients were significantly different between the two groups ( P<0.05). Multivariate logistic regression analysis found BMI≥25, intracapsular invasion, extrathyroidal invasion, lymph node metastasis, proportion of PT1 patients were independent factors affecting Tg in patients with PTC (95% CI=3.845-11.735, 1.485-2.983,1.171-2.762,4.083-16.526,1.003-2.174, P<0.05). There was a negative correlation between the expression of TPO and Tg in PTC ( r=-0.498, P<0.001) . Conclusion:TPO and Tg are highly correlated with tumor lymphatic metastasis, pathological grade, tumor diameter and tumor invasion range in patients with papillary thyroid carcinoma, and the expression of the two is negatively correlated, which can be used as effective indicators for evaluating the prognosis of patients.

Role and prospect of ferroptosis in ischemia-reperfusion injury during organ transplantation / 器官移植

Ferroptosis is a newly-emerged pattern of programmed cell death discovered in recent years, which is defined as iron-dependent programmed necrosis mediated by lipid peroxidation damage. As a conservative procedure, ferroptosis plays a vital role in the development and diseases of multiple organisms including plants and animals. Since ferroptosis was first reported in 2012, growing interests have been diverted to the process of ferroptosis and its role in disease treatment. Ischemia-reperfusion injury is a common pathological process during organ transplantation, and ferroptosis is considered as one of the main patterns inducing ischemia-reperfusion injury. Consequently, the definition, regulatory mechanism and the mechanisms of ferroptosis in ischemia-reperfusion injury after kidney, liver, heart and lung transplantations were reviewed, aiming to provide theoretical basis for the prevention and treatment of ischemia-reperfusion injury in organ transplantation.

Determination of Serum Unbound Bilirubin and Its Clinical Application Value in Neonates / 中山大学学报(医学科学版)

ObjectiveTo investigate the clinical appropriateness and application value of the peroxidase （POD） method for the detection of unbound bilirubin （UB） in neonatal serum. MethodsHydrogen peroxide （0.33 mol/L） and three different final concentrations （0.019， 0.038， 0.075 μg/mL） of horseradish peroxidase （HRP） were added to standard bilirubin solution （1， 2， 3 μmol/L） to obtain a standardized HRP primary rate constant Kp. Then 25 μL of neonatal serum was diluted by 41.6 fold， and measured with 2.4 and 4.8 μg/mL HRP at 37 ℃ under the dark， to determine the UB concentration. The accuracy， precision， and stability of the methodology were validated. The clinical characteristics of 33 jaundiced neonates were collected， including total serum bilirubin （TSB）， indirect bilirubin （IDB）， albumin （ALB）， bilirubin to albumin molar ratio （BAMR）， etc. The experimental data were analyzed by Graphpad Prism 8.0. ResultsA standardized Kp of （7.20±1.08） mL·μg-1·min-1 was determined at pH 7.4±0.2， 37 ℃ in the dark. The HRP activity and UB concentrations remained stable at -20 ℃ for 3 weeks and a week， respectively. The mean intra-day and inter-day coefficients of variation of the serum samples with different UB concentrations were less than 10%. In this study， the UB concentrations in 33 jaundiced neonates （gestational age ≥35 weeks） were measured by the POD method in the range of （0.32～1.20） μg/dL， which was positively correlated with TSB， IDB and BAMR. Of the five infants whose UB concentrations measured more than 1 μg/dL， three received intensive phototherapy （60%）. ConclusionsThe POD method combined with a standard equipment spectrophotometer to detect serum UB concentrations in neonates is easy to operate， rapid to detect， and low cost. This method has good accuracy and precision， which is convenient for clinical implementation. Moreover， the measurement of serum UB may assist us in better management of neonatal jaundice in clinical practice.

Mechanism of blue light-induced ferroptosis in retinal pigment epithelium cells / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the occurrence and possible mechanism of blue light-induced ferroptosis in retinal pigment epithelial cells.METHODS: ARPE-19 cells cultured in vitro were irradiated by 405 nm blue light at 50 mW/cm2 irradiance with different duration and were divided into control, 16.3J/cm2, 32.6J/cm2, and 65.2J/cm2 groups; the 65.2J/cm2 group was defined as the high-level blue light irradiation group and cells were further divided into control, high-level blue light irradiation group and high-level blue light irradiation + ferroptosis inhibitor group. CCK-8 assay was used to detect cell viability, commercial kits were used to detect intracellular glutathione(GSH), ferrous iron and malondialdehyde(MDA)concentration, and Western blot was used to detect the relative expression of glutathione peroxidase 4(GPX4)and xCT proteins in cells.RESULTS: The decrease of ARPE-19 cell viability caused by blue light irradiation was dose-dependent, and the reduction of intracellular GSH concentration, the increase of ferrous iron concentration and MDA concentration were all caused by high-level blue light irradiation(all P&#x0026;#x003C;0.05); the ferroptosis inhibitor partially restored cell viability and recovered intracellular GSH, reduced concentrations of MDA and ferrous iron in the blue light irradiation group(all P&#x0026;#x003C;0.05). The relative expressions of GPX4 and xCT proteins were significantly decreased in the blue light irradiation group, and such change was alleviated by the ferroptosis inhibitor(P&#x0026;#x003C;0.05).CONCLUSION: Blue light irradiation may induce ferroptosis in RPE cells by targeting the xCT and GPX4-associated antioxidant pathways.

In vivo antioxidant activity of rabbiteye blueberry (Vaccinium ashei cv. 'Brightwell') anthocyanin extracts / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Blueberries are rich in phenolic compounds including anthocyanins which are closely related to biological health functions. The purpose of this study was to investigate the antioxidant activity of blueberry anthocyanins extracted from 'Brightwell' rabbiteye blueberries in mice. After one week of adaptation, C57BL/6J healthy male mice were divided into different groups that were administered with 100, 400, or 800 mg/kg blueberry anthocyanin extract (BAE), and sacrificed at different time points (0.1, 0.5, 1, 2, 4, 8, or 12 h). The plasma, eyeball, intestine, liver, and adipose tissues were collected to compare their antioxidant activity, including total antioxidant capacity (T-AOC), superoxide dismutase (SOD) activity and glutathione-peroxidase (GSH-PX/GPX) content, and the oxidative stress marker malondialdehyde (MDA) level. The results showed that blueberry anthocyanins had positive concentration-dependent antioxidant activity in vivo. The greater the concentration of BAE, the higher the T-AOC value, but the lower the MDA level. The enzyme activity of SOD, the content of GSH-PX, and messenger RNA (mRNA) levels of Cu,Zn-SOD, Mn-SOD, and GPX all confirmed that BAE played an antioxidant role after digestion in mice by improving their antioxidant defense. The in vivo antioxidant activity of BAE indicated that blueberry anthocyanins could be developed into functional foods or nutraceuticals with the aim of preventing or treating oxidative stress-related diseases.

Novel perspective in transplantation therapy of mesenchymal stem cells: targeting the ferroptosis pathway / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Ex vivo culture-amplified mesenchymal stem cells (MSCs) have been studied because of their capacity for healing tissue injury. MSC transplantation is a valid approach for promoting the repair of damaged tissues and replacement of lost cells or to safeguard surviving cells, but currently the efficiency of MSC transplantation is constrained by the extensive loss of MSCs during the short post-transplantation period. Hence, strategies to increase the efficacy of MSC treatment are urgently needed. Iron overload, reactive oxygen species deposition, and decreased antioxidant capacity suppress the proliferation and regeneration of MSCs, thereby hastening cell death. Notably, oxidative stress (OS) and deficient antioxidant defense induced by iron overload can result in ferroptosis. Ferroptosis may inhibit cell survival after MSC transplantation, thereby reducing clinical efficacy. In this review, we explore the role of ferroptosis in MSC performance. Given that little research has focused on ferroptosis in transplanted MSCs, further study is urgently needed to enhance the in vivo implantation, function, and duration of MSCs.

Resveratrol Attenuates Degeneration and Apoptosis of Cardiomyocytes Induced by Aortic Clamping

ABSTRACT Introduction: Objective: To investigate the potential beneficial effects of resveratrol (RVT) against ischemia-reperfusion injury of myocardial tissue during surgical treatment of ruptured abdominal aortic aneurysm. Methods: Four groups were established — control, ischemia/reperfusion (I/R), sham (I/R+solvent/dimethyl sulfoxide [DMSO]), and I/R+RVT. Ruptured abdominal aortic aneurysm model was used as the experimental protocol. Results: In the I/R and I/R+DMSO groups, malondialdehyde (MDA) levels in myocardial tissue were found to be significantly increased compared to the control group. The MDA level in myocardial tissue was significantly decreased in the I/ R+RVT group compared to the I/R group. In I/R and I/R+DMSO groups, glutathione peroxidase (GSH) levels in myocardial tissue were found to be significantly decreased compared to the control group. The GSH level in the myocardial tissue was significantly increased in the I/R+RVT group compared to the I/R group. In the light microscope, isotropic and anisotropic band disorganized atypical cardiomyocytes in the I/R group and degenerative cardiomyocytes and edematous areas in the I/R+DMSO group were observed. Degenerative cardiomyocytes and edematous areas were decreased in the I/R+RVT group. When heart tissue sections incubated with cleaved caspase-3 primary antibodies were examined under the light microscope, apoptotic cardiomyocytes were present in I/R and I/R+DMSO groups. A decrease in the number of apoptotic cardiomyocytes was observed in the I/R+RVT group. Conclusion: The findings of the present study indicate that RVT exhibits protective effects against ischemia-reperfusion injury occurring in the myocardium as a distant organ as a result of abdominal aorta clamping.

Analysis of changes in routine blood parameters and thyroid hormone levels in women at different stages of pregnancy and their correlations with TPOAb / 上海预防医学

ObjectiveTo investigate the patterns of changes in routine blood parameters， thyroid hormone levels， and their correlations with thyroid peroxidase antibodies （TPOAb） among women at different stages of pregnancy， so as to provide a theoretical basis for maternal and child health care and diagnosis and treatment. MethodsA total of 732 pregnant women at different stages of pregnancy who underwent health examinations at the First Maternity and Infant Hospital affiliated to Tongji University from May 2020 to August 2022 were selected as the observation group. The group comprised 245 women in the first trimester （≤12 weeks）， 247 women in the second trimester （between13 and 27 weeks） and 240 women in the third trimester （≥28 weeks）. Additionally， 240 non-pregnant， healthy women of child-bearing age who conducted their health checkups in the same hospital were selected as the control group. All the research subjects were required to provide peripheral venous blood samples to measure hemoglobin （Hb）， standard deviation of red blood cell distribution width （RDW-SD）， coefficient of variation of red blood cell distribution width （RDW-CV）， platelet （Plt） count， platelet distribution width （PDW）， as well as thyroid stimulating hormone （TSH）， total thyroxine （TT4）， total triiodothyronine （TT3）， free thyroxine （FT4）， free triiodothyronine （FT3）， and TPOAb. The results were statistically analyzed. ResultsWith advancing gestational age， Hb levels were significantly lower in the second and third trimesters than in the first trimester and the control group （F=68.25， P<0.001）， while RDW-SD and RDW-CV were significantly higher （F=41.34， P<0.001； F=3.64， P=0.012）. Plt levels throughout pregnancy were significantly lower than that in the control group （F=43.21， P<0.001）. TSH levels were significantly lower in the first and second trimesters than in the control group （Z=53.49， P<0.001）， but gradually increased with gestational age. TT3 and TT4 levels were significantly higher than those in the control group throughout pregnancy （F＝148.25， P<0.001； F＝210.83， P<0.001）， while FT3 and FT4 levels were significantly lower in the second and third trimesters than in the first trimester and the control group （F=42.95， P<0.001； F=101.73， P<0.001）. The abnormal rate of TPOAb was significantly higher than that in the control group throughout pregnancy （χ2＝25.61， P<0.001）. Among pregnant women， those with TPOAb positivity had significantly higher TSH levels and RDW-CV than those with TPOAb negativity （Z＝5.70， P<0.001； t＝2.39， P＝0.018）. ConclusionThe levels of Hb， Plt， and thyroid hormones in pregnant women are closely related to gestational age. With increasing gestational age， the abnormal rate of TPOAb decreases， but the TSH levels and RDW-CV of TPOAb positive pregnant women are higher， requiring clinical attention and screening to improve maternal and child health.

Expressions of serum thyroid stimulating hormone, thyroid peroxidase antibody and thyroglobulin antibody in papillary thyroid carcinoma and their clinical significances / 肿瘤研究与临床

Objective:To explore the relationship between serum levels of thyroid stimulating hormone (TSH), thyroid peroxidase antibody (TPO-Ab) and thyroglobulin antibody (Tg-Ab) and the development of papillary thyroid carcinoma.Methods:The clinical data of 574 patients with thyroid nodules who received surgical treatment in Tengzhou Central People's Hospital from January to December 2021 were retrospectively analyzed. Using histopathological diagnosis as the gold standard, the patients were divided into papillary thyroid carcinoma group (malignant group, 267 cases) and benign thyroid nodules group (benign group, 307 cases). The clinical data and the preoperative serum TSH, TPO-Ab and Tg-Ab levels were compared between the two groups. The correlation between preoperative serum TSH, TPO-Ab and Tg-Ab levels and papillary thyroid cancer was analyzed by logistic regression.Results:There were no statistical differences in the age and gender of patients between the malignant group and the benign group (all P > 0.05). TSH [2.37 mIU/L (1.43 mIU/L, 5.09 mIU/L)], TPO-Ab [17.84 IU/ml (11.94 IU/ml, 40.68 IU/ml)] and Tg-Ab [15.69 IU/ml (10.57 IU/ml, 132.00 IU/ml)] in the malignant group were higher than those in the benign group [TSH 1.60 mIU/L (0.88 mIU/L, 2.57mIU/L), TPO-Ab 14.29 IU/ml (10.00 IU/ml, 21.17 IU/ml), Tg-Ab 12.23 IU/ml (10.00 IU/ml, 16.51 IU/ml)], and the differences were statistically significant ( Z values were -6.43, -4.60 and -6.15, all P < 0.05). Multivariate logistic regression analysis showed that positive TPO-Ab ( OR = 0.996, 95% CI 0.993-0.999, P = 0.013) and positive Tg-Ab ( OR = 0.996, 95% CI 0.994-0.998, P < 0.05) were independent risk factors for papillary thyroid cancer. Conclusions:Preoperative serum TSH, TPO-Ab and Tg-Ab levels are closely related to papillary thyroid cancer, among which positive serum TPO-Ab and positive Tg-Ab are independent risk factors for papillary thyroid cancer and have important values in the differential diagnosis of benign and malignant thyroid nodules.

Research progress on the role and mechanism of ferroptosis in renal fibrosis / 中华肾脏病杂志

Ferroptosis is a recently identified type of non-apoptotic cell death, mainly caused by disruption of cellular metabolic pathways such as iron metabolism and reactive oxygen species metabolism, characterized by intracellular iron overload and reactive oxygen species accumulation leading to lipid peroxidation. Ferroptosis is closely related to renal diseases. The role of ferroptosis in diseases such as acute kidney injury and renal cell carcinoma has been extensively studied, and new discoveries and advances have been made in its relationship with renal fibrosis. The paper systematically reviews the relationship between ferroptosis and renal fibrosis in terms of the latest regulatory mechanisms of ferroptosis and its role in renal fibrosis, and explores the potential clinical application of targeted inhibition of ferroptosis to prevent renal fibrosis.

Correlation between food-specific IgG antibodies and phenotypes of chronic spontaneous urticaria / 中华皮肤科杂志

Objective:To investigate the correlation between food-specific IgG (sIgG) antibodies and phenotypes of chronic spontaneous urticaria (CSU) .Methods:Serum samples were collected from outpatients with active CSU, symptomatic dermographism (SD) , or acute urticaria (AU) , and healthy controls from 5 third-grade class-A hospitals such as the First Hospital of China Medical University between April 2014 and March 2015. Enzyme-linked immunosorbent assay was conducted to detect serum levels of 90 food-sIgG antibodies and total IgE, Western blot analysis to detect levels of 20 allergen-specific IgE antibodies, and chemiluminescent microparticle immunoassay to detect levels of anti-thyroid peroxidase IgG antibodies and anti-thyroglobulin IgG antibodies. Comparisons of normally distributed quantitative data between two groups and among several groups were performed by t test and one-way analysis of variance, respectively; comparisons of non-normally distributed quantitative data between two groups were performed by Mann-Whitney U test; for comparisons of proportions, chi-square test and Fisher′s exact test were used. Results:A total of 248 patients with CSU, 22 with SD, 15 with AU and 13 healthy controls were recruited. The cut-off level for sIgG positivity was 100 U/ml (at least 2+) , and the positive rate of food-sIgG antibodies was slightly higher in the patients with CSU (176/248, 70.97%) , SD (15/22, 68.18%) and AU (11/15) than in the healthy controls (7/13; χ2 = 1.80, P = 0.615) . Among the 248 CSU patients, the proportion of patients with family history of allergic diseases was significantly higher in the sIgG-positive group (71/176, 40.34%) than in the sIgG-negative group (19/72, 26.39%; χ2 = 4.30, P = 0.042) , while no significant difference was observed in the 1-day urticaria activity score (UASday) between the two groups ( Z = 0.18, P = 0.859) . Totally, 177 CSU patients completed 12- to 40-week treatment; their condition could be completely controlled by second-generation H1-antihistamines, and there was no significant difference in the required dosage of second-generation H1-antihistamines between the sIgG-positive group (128 cases) and sIgG-negative group (49 cases; Z = -1.06, P = 0.298) . Conclusions:The prevalence of family history of allergic diseases was relatively high in food-sIgG-positive patients with CSU. However, food-sIgG could not be used as an indicator to reflect the disease activity of CSU and treatment response.

Role of Nrf2/GPX4 signaling pathway-mediated ferroptosis in midazolam-induced reduction of HIBD in neonatal rats / 中华麻醉学杂志

Objective:To evaluate the role of nuclear factor-erythroid 2-related factor 2 (Nrf2)/glutathione peroxidase-4 (GPX4) signaling pathway-mediated ferroptosis in midazolam-induced reduction of hypoxic-ischemic brain damage (HIBD) in neonatal rats.Methods:Ninety healthy 7-day-old neonatal rats, weighing 16-20 g, were divided into 6 groups ( n=15 each) using the random number table method: sham operation group (Sham group), HIBD group, low-dose midazolam (10 mg/kg) group (group L), medium-dose midazolam (20 mg/kg) group (group M), high-dose midazolam (40 mg/kg) group (group H), and Nrf2 inhibitor ML385 group (group I). The HIBD model was developed by ligating the left carotid artery and exposing to a hypoxic condition for 2 h in anesthetized animals. Starting from 2nd day after developing the model, the corresponding doses of midazolam were intraperitoneally injected in midazolam groups, the equal volume of normal saline was intraperitoneally injected in Sham and HIBD groups, midazolam 40 mg/kg and Nrf2 inhibitor ML385 30 mg/kg were intraperitoneally injected once a day for 8 consecutive days in group I. The rats were weighed and subjected to the Morris water maze test after the end of administration. Blood samples were taken from the abdominal aorta after the end of the Morris water maze test, and then the animals were sacrificed to remove the brain for determination of the concentrations of serum iron, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) (by enzyme-linked immunosorbent assay), contents of iron and GSH in hippocampal tissues (by ultraviolet spectrophotometry and micro method), the number of Nrf2/neuronal nuclear antigen (NeuN) and GPX4/NeuN positive cells (by immunofluorescent staining), and expression of Nrf2, GPX4, and 4-hydroxynonaenoic acid (4-HNE) in hippocampal tissues and for microscopic examination of the pathological changes of hippocampal neurons in brain tissues (after HE staining and Nissl staining). Results:Compared with Sham group, the first time to arrival at platform was significantly prolonged, the number of crossing the origional platform was reduced, and the time of staying at the target quadrant was shortened, the iron content in the hippocampal tissues was increased, the content of GSH and the number of Nrf2/NeuN and GPX4/NeuN positive cells were decreased, the expression of Nrf2 and GPX4 was down-regulated, the expression of 4-HNE was up-regulated, the concentrations of serum iron, IL-6 and TNF-α were increased, and the injury to hippocampal neurons was marked in HIBD group ( P<0.05). Compared with HIBD group, the first time to arrival at platform was significantly shortened, the number of crossing the origional platform was increased, and the time of staying at the target quadrant was prolonged, the iron content in the hippocampus tissues was decreased, the content of GSH and the number of Nrf2/NeuN and GPX4/NeuN positive cells were increased, the expression of Nrf2 and GPX4 was up-regulated, the expression of 4-HNE was down-regulated, the concentrations of serum iron, IL-6 and TNF-α were decreased ( P<0.05), and the injury to hippocampal neurons was significantly reduced in H, M and L groups. Compared with group H, the first time to arrival at platform was significantly prolonged, the number of crossing the origional platform was reduced, and the time of staying at the target quadrant was shortened, the iron content in the hippocampus tissue was increased, the content of GSH and the number of Nrf2/NeuN and GPX4/NeuN positive cells were decreased, the expression of Nrf2 and GPX4 was down-regulated, the expression of 4-HNE was up-regulated, the concentrations of serum iron, IL-6 and TNF-α were increased ( P<0.05), and the injury to hippocampal neurons was aggravated in group I. Conclusions:The mechanism by which midazolam reduces HIBD may be related to activation of the Nrf2/GPX4 signaling pathway and inhibition of hippocampal neuronal ferroptosis in neonatal rats.

Effect of diosmetin on ferroptosis of hippocampal cells in rats with bacterial meningitis and the mechanism of SIRT1-Nrf2 signaling pathway / 中华行为医学与脑科学杂志

Objective:To explore the effect and mechanism of diosmetin (Dio) on neuronal ferroptosis in rats with bacterial meningitis (BM).Methods:Male SD rats aged 6-7 weeks of SPF grade were selected for the experiment. The BM model was established by injecting group B hemolytic streptococcus into the cisterna magna of cerebellum. Sixty BM model rats were successfully modeled and divided into model group, low-dose Dio group, medium-dose Dio group, high-dose Dio group and inhibitor group according to the random number table method, with 12 rats in each group. Another 12 weight-matched rats were taken as the control group.The rats in the low-dose Dio group, medium-dose Dio group, high-dose Dio group and the inhibitor group were intragastrically administered with Dio at 50 mg/kg, 100 mg/kg, 200 mg/kg and 200 mg/kg, respectively. The rats in the control group were intragastrically administered with an equal volume of 0.9 % sodium chloride solution. On the day of intragastric administration, the rats in the inhibitor group were intraperitoneally injected with SIRT1 pathway inhibitor EX527 (10 mg/kg), and the rats in the other groups were injected with an equal volume of 0.9% sodium chloride solution. The above interventions were performed once a day for 28 consecutive days. Loeffler neurological score was used to evaluate the neurological impairment in rats. Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in cerebrospinal fluid of rats were detected by ELISA. The number of white blood cells in cerebrospinal fluid was detected by a blood cell analyzer. Glutathione (GSH) was detected by micro-enzyme labeling method, malondialdehyde (MDA) was detected by thiobarbituric acid colorimetric method, reactive oxygen species(ROS) was detected by colorimetry, and Fe 2+ level was detected by ferrozine method. Hematoxylin-eosin staining, Prussian blue staining and TUNEL staining were used to observe the pathological damage, iron accumulation and apoptosis in the hippocampus, respectively.Western blot was applied to measure the expression of transferrin (Tf), proliferating cell nuclear antigen (PCNA), Bcl-2-associated X protein (Bax), caspase-3 and SIRT1/Nrf2/HO-1/Gpx4 signaling pathway proteins. Graphpad Prism 9.0 was used for data analysis. One-way ANOVA was used for statistical analysis, and SNK- q test was used for further pairwise comparisons. Results:(1) There was a statistically significant difference in neurological function scores among the 6 groups of rats ( F=125.451, P<0.001). The neurological function score of the model group was lower than that of control group, while the neurological function scores of the low-dose Dio group, medium-dose Dio group, and high-dose Dio group were higher than those of the model group (all P<0.05). The neurological function score of the inhibitor group ((2.57±0.26)) was lower than that of high-dose Dio group ((4.34±0.48)) ( P<0.05). (2) There were statistically significant differences in the levels of IL-6, TNF-α and the number of white blood cells in the cerebrospinal fluid of rats among the 6 groups ( F=127.817, 102.413, 180.967, all P<0.001). The levels of IL-6, TNF-α and the number of white blood cells in model group were higher than those of control group(all P<0.05). The levels of IL-6, TNF-α and the number of white blood cells in low-dose Dio group, medium-dose Dio group and high-dose Dio group were lower than those of model group (all P<0.001), and those in inhibitor group were all higher than those in high-dose Dio group(all P<0.001). (3) There were statistically significant differences in iron deposition rate and neuronal apoptosis rate among the 6 groups of rats ( F=90.857, 88.835, both P<0.001). The iron deposition rate ((18.37±3.14)%) and neuronal apoptosis rate ((27.58±2.63)%) in the inhibitor group were higher than those in the high-dose Dio group ((6.35±1.08)%, (14.02±1.87)%) (both P<0.05). (4) The levels of GSH, ROS, MDA, and Fe 2+ in the hippocampus of the 6 groups of rats showed statistically significant differences ( F=54.465, 106.453, 55.969, 105.457, all P<0.001). The GSH content in the inhibitor group ((103.48±8.76) mmol/g) was lower than that in the high-dose Dio group ((133.97±10.54) mmol/g), while the contents of ROS, MDA, Fe 2+ ((225.17±16.32) μmol/mg, (10.73±1.58) μmol/mg, (62.71±5.43) μg/g) were higher than those of the high-dose Dio group ((131.87±11.67) μmol/mg, (4.35±0.87) μmol/mg, (34.86±2.95) μg/g) (all P<0.05). (5)There were statistically significant differences in the protein levels of Tf, PCNA, Bax, caspase-3, SIRT1, Nrf2, HO-1 and Gpx4 in the hippocampus of the 6 groups of rats ( F=120.179, 107.568, 157.265, 98.031, 90.932, 52.283, 59.424, 114.539, all P<0.001). The protein levels of Tf, Bax and caspase-3 in the hippocampus of inhibitor group were higher than those of the high-dose Dio group, while the protein levels of PCNA, SIRT1, Nrf2, HO-1, Gpx4 were lower than those of the high-dose Dio group (all P<0.05). Conclusion:Diosmetin can activate SIRT1/Nrf2/HO-1/Gpx4 signaling pathway, thereby inhibiting neuronal ferroptosis in BM rats.

Effects of neuroendoscopic hematoma removal versus soft channel drainage in the treatment of chronic subdural hematoma / 中国基层医药

Objective:To investigate the clinical efficacy of neuroendoscopic hematoma removal versus soft channel drainage in the treatment of chronic subdural hematoma. Methods:The clinical data of 102 patients with chronic subdural hematoma who received treatment in Jincheng People's Hospital from May 2018 to May 2020 were retrospectively analyzed. They were divided into the neuroendoscopy group ( n = 50) and the soft channel group ( n = 52) according to different surgical methods. Perioperative indexes, hematoma clearance rate, China Stroke Scale score, the activity of daily living score, and oxidative stress indexes were compared between the two groups. All patients were followed up for 3 months. The incidence of complications during the follow-up period was calculated. Results:The retention time of the drainage tube in the neuroendoscopy group was shorter than that in the soft channel group [(2.45 ± 0.63) days vs. (3.30 ± 0.78) days, t = 6.06, P < 0.001]. The length of hospital stay in the neuroendoscopy group was shorter than that in the soft channel group [(7.14 ± 1.65) days vs. (9.07 ± 2.11) days, t = 5.15, P < 0.001]. The hematoma clearance rate at postoperative 7 days in the neuroendoscopy group was higher than that in the soft channel group [(93.45 ± 5.50)% vs. (81.86 ± 7.24)%, χ2 = 9.12, P < 0.001]. There were no significant differences in operation time and intraoperative blood loss between the two groups (both P > 0.05). At postoperative 30 days, the China Stroke Scale score in the neuroendoscopy group was lower than that in the soft channel group [(12.74 ± 2.23) points vs. (18.67 ± 2.45) points, t = 12.79, P < 0.001]. The activity of daily life score in the neuroendoscopy group was significantly higher than that in the soft channel group [(77.69 ± 7.11) points vs. (91.35 ± 7.25) points, t = 9.60, P < 0.001]. At postoperative 7 days, glutathione peroxidase level in the neuroendoscopy group was significantly lower than that in the soft channel group [(130.75 ± 13.66) U/L vs. (148.60 ± 14.64) U/L, t = 6.37, P < 0.001]. Malondialdehyde level in the neuroendoscopy group was significantly lower than that in the soft channel group [(5.11 ± 0.65) nmol/L vs. (6.19 ± 0.74) nmol/L, t = 7.83, P < 0.001]. Superoxide dismutase level in the neuroendoscopy group was significantly higher than that in the soft channel group [(275.60 ± 22.33) U/L vs. (254.60 ± 18.55) U/L, t = 5.15, P < 0.001]. There was no significant difference in the incidence of complications between the two groups ( P > 0.05). Conclusion:Compared with soft channel drainage, neuroendoscopic hematoma removal can obtain better short-term curative effects and less oxidative stress response in the treatment of chronic subdural hematoma. Neuroendoscopic hematoma removal does not increase the incidence of postoperative complications and is highly safe.

Vitamin D and autoimmune thyroiditis in children – A prospective case–control study

Background: The deficiency or insufficiency of Vitamin D has been widely reported to be linked with autoimmune thyroid diseases. Several studies were evaluated the immunomodulatory effects of 25 hydroxyvitamin D (25(OH)D) and its counterparts in autoimmune diseases especially in autoimmune thyroiditis. Aim and Objectives: The aim of the study was to assess the Vitamin D status in children with autoimmune thyroiditis at tertiary care hospital, Sangareddy, Telangana. Materials and Methods: A source of 80 newly diagnosed cases with autoimmune thyroiditis and similar volume of age and sex matched control subjects between ?6 and 12 years were included in the study. Parameters such as thyroid function tests, serum calcium, serum phosphorus, serum alkaline phosphatase, 25(OH)D, and antithyroid antibodies levels were assessed. The antithyroid antibodies levels were assessed through chemiluminescence assay. Results: The 25(OH)D levels were 14.98ng/ml in cases and 17.46 ng/ml in control subjects. The mean levels of 25(OH)D, serum calcium, and alkaline phosphatase were statistically significant (P < 0.05). Conclusion: The levels of Vitamin D and four groups of antithyroid peroxidase antibody and antithyroglobulin antibody among cases and control subjects were not significant (P > 0.05). The estimation of Vitamin D in high-risk group may be helpful in designing the treatment strategies to decrease the morbidity.