Detection of Amantadine by Label-free Fluorescence Method Based on Truncated Aptamer and Molybdenum Disulfide Nanosheet Signal Enhancement Strategy / 分析化学

Amantadine(AMD)residue can accumulate in organisms through the food chain and cause serious harm to human body.AMD can specifically bind to AMD specific aptamer and cause its conformation to change from a random single strand to a stem-loop structure.To avoid the influence of excess nucleotides on binding of aptamer to AMD,the truncation of the AMD original aptamer J was optimized by retaining an appropriate stem-loop structure,and a new type of truncation aptamers was developed in this work.By comparing the truncated aptamer with the original aptamer,it was found that the truncated aptamer J-7 had better affinity and specificity with AMD.The detection limit of AMD was 0.11 ng/mL by using J-7 as specific recognition element and molybdenum disulfide nanosheet(MoS2Ns)as signal amplification element.The developed method base on truncated aptamer J-7 was used for detection of AMD in milk,yogurt and SD rat serum samples for the first time with recoveries of 86.6%-108.2%.This study provided a reference for truncating other long sequence aptamers and provided a more sensitive detection method for monitoring AMD residues in food.

Regulation of proliferation and invasion of renal cancer cells by miRNA-4469 via targeting PDIA4 gene / 肿瘤研究与临床

Objective:To explore the mechanism by which miRNA-4469 (miR-4469) regulates the proliferation and invasion of renal cancer cells in vitro.Methods:The survival differences of patients with different expression levels of miR-4469 were analyzed based on the OncomiR database. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) method was used to detect the expression of miR-4469 in renal cancer cell lines ACHN, OS-RC-2, SK-RC-20, 769-P, A498 and normal renal tubular epithelial cell line HK-2, and the renal cancer cells with the lowest expression level of miR-4469 were divided into miR-4469 group and control group, and were transfected with miR-4469 mimic and negative control sequence, respectively. The CCK-8 assay was used to detect the cell proliferation ability (expressed as absorbance value) in the two groups, and Transwell assay was used to analyze the number of invasive cells in the two groups. TargetScan Release 8.0 software was used to predict the binding site between miR-4469 and protein disulfide isomerase A4 (PDIA4) mRNA, and dual-luciferase reporter gene assay was used to verify the targeting relationship between miR-4469 and PDIA4 mRNA. qRT-PCR method was used to detect the expression of PDIA4 mRNA in cells of each group, and Western blotting method was used to detect the expression levels of PDIA4 protein and PI3K-AKT-m-TOR pathway proteins in cells of each group.Results:Analysis of relevant data from the OncomiR database showed that compared with patients with low miR-4469 expression, the overall survival of renal cancer patients with high miR-4469 expression was better ( P < 0.001). The relative expression of miR-4469 in each renal cancer cell line was lower than that in HK-2 cells (all P < 0.05), and the expression of miR-4469 in 769-P cells was the lowest, which were selected to perform the subsequent experiments. The proliferation ability of 769-P cells in the miR-4469 group was lower than that in the control group ( P < 0.01). The number of 769-P cell invasions in the miR-4469 group were less than that in the control group [(19±3) cells vs. (64±7) cells, t = 5.44, P = 0.002]. Compared with the co-transfection of wild-type PDIA4 and miR-4469 negative sequence group, the relative luciferase activity of cells in the co-transfection of wild-type PDIA4 and miR-4469 mimic sequence group was lower (0.42±0.07 vs. 1.01±0.08, t = 5.74, P = 0.001); there was no statistical difference in cell luciferase activity between the co-transfected mutant PDIA4 and miR-4469 negative sequence group and the co-transfected mutant PDIA4 and miR-4469 mimic sequence group (0.99±0.11 vs. 1.02±0.11, t = 0.19, P = 0.001). The relative expression levels of PDIA4 mRNA in 769-P cells in the miR-4469 group were lower than that in the control group (0.98±0.23 vs. 7.19±2.23, t = 2.77, P = 0.032). Compared with the control group, the expression of PDIA4 protein and PI3K-AKT-m-TOR pathway-related p-PI3K, p-AKT, p-mTOR, and p-SGK1 proteins in 769-P cells in the miR-4469 group were all lower (all P < 0.05). Conclusions:miR-4469 may be related to the survival of renal cancer patients, and its expression is down-regulated in various renal cancer cell lines. miR-4469 may inhibit the proliferation and invasion of renal cancer 769-P cells by regulating the PI3K-AKT-m-TOR pathway through PDIA4.

Diallyl disulfide augments sensitivity of DJ-1 overexpressed human gastric cells to 5-FU / 中国药理学通报

Aim To investigate whether diallyl disul-fide (DADS) augments the sensitivity of DJ-1 (protein/ nucleic acid deglycase) overexpressed human gastric SGC7901 cells to 5-FU (5-fluorouracil). Methods The experimental groups include control group, DADS group, VCR (vincristine) group, VCR + DADS group, DJ-1 group, DJ-1 + DADS group. MTT was used to analyze the effect of DADS on 5 -FU (5 -fluorou- racil) induced proliferation inhibition. Flow cytometry was performed to examine the effect of DADS on cell apoptosis. RT-PCR, Western blot, and immunofluo-rescence were used for determine the effect of DADS on the drug resistance associated gene expression. Results DADS enhanced the proliferation inhibitory effect of 5-FU on DJ-1 overexpressed cells and VCR resistant cells. DADS could induce apoptosis in VCR-resistant cells. DADS downregulated the expression of DJ-1 while inducing apoptosis in DJ-1 overexpressed cells. DJ-1 overexpression upregulated the expression of P-gp (P-glycoprotein), Bcl-2, and XIAP (X-linked inhibitor of apoptosis protein), downregulated the expression of caspase-3. DADS decreased the expression of P-gp, Bcl-2, and XIAP, while increased the expression of caspase-3 in DJ-1 overexpressed cells and VCR-resistant cells. Conclusions DADS can augment the sensitivity of DJ-1 overexpressed cells to 5-FU, which is related to its antagonism against DJ-1 mediated upregula- tion of P-gp, Bcl-2, XIAP, and downregulation of caspase-3.

Did diet compliance and remission reduce oxidative stress in celiac patients?

SUMMARY OBJECTIVE: We aimed to examine the effect of remission status on thiol-disulfide homeostasis in celiac patients and thus to indirectly determine the effect of oxidative stress and inflammation caused by non-compliance with the diet. METHODS: Between February 2019 and December 2021, 117 patients diagnosed with celiac disease were included in this prospective randomized and controlled study. In addition to routine tests of celiac patients, thiol and disulfide measurements were made from the blood both at the beginning of the study and at the end of the first year. RESULTS: While 52 of the patients (44.4%) were in remission, 65 patients (55.6%) were not. There was an evident increase in native thiol levels of the patients who were initially not in remission but went into at the end of the first year (347.4±46.7 μmol/L vs. 365.3±44.0 μmol/L; p=0.001). Mean plasma disulfide levels of patients with celiac going into remission became reduced in the first year from the level of 14.5±5.1 μmol/L down to 8.9±4.2 μmol/L (p<0.001). In celiac patients who entered remission, disulfide and anti-tissue transglutaminase immunoglobulin A levels decreased in a correlation (r=0.526; p<0.001). CONCLUSION: Not being in remission in celiac disease leads to increased oxidative stress, and thiol-disulfide homeostasis is an indirect indicator of this. Additionally, providing remission in celiac patients reduces oxidative stress.

Downregulated miR-181a alleviates H2O2-induced oxidative stress and cellular senescence by targeting PDIA6 in human foreskin fibroblasts

Abstract Background Oxidative stress is strongly associated with cellular senescence. Numerous studies have indicated that microRNAs (miRNAs) play a critical part in cellular senescence. MiR-181a was reported to induce cellular senescence, however, the potential mechanism of miR-181a in hydrogen peroxide (H2O2)-induced cellular senescence remains obscure. Objective The aim of this study is to investigate the role and regulatory mechanism of miR-181a in H2O2-induced cellular senescence. Methods Human foreskin fibroblasts (HFF) transfected with miR-181a inhibitor/miR-NC with or without H2O2 treatment were divided into four groups: control + miR-NC/miR-181a inhibitor, H2O2 + miR-NC/miR-181a inhibitor. CCK-8 assay was utilized to evaluate the viability of HFF. RT-qPCR was used to measure the expression of miR-181a and its target genes. Protein levels of protein disulfide isomerase family A member 6 (PDIA6) and senescence markers were assessed by western blotting. Senescence-associated β-galactosidase (SA-β-gal) staining was applied for detecting SA-β-gal activity. The activities of SOD, GPx, and CAT were detected by corresponding assay kits. The binding relation between PDIA6 and miR-181a was identified by luciferase reporter assay. Results MiR-181a inhibition suppressed H2O2-induced oxidative stress and cellular senescence in HFF. PDIA6 was targeted by miR-181a and lowly expressed in H2O2-treated HFF. Knocking down PDIA6 reversed miR-181a inhibition-mediated suppressive impact on H2O2-induced oxidative stress and cellular senescence in HFF. Study limitations Signaling pathways that might be mediated by miR-181a/PDIA6 axis were not investigated. Conclusion Downregulated miR-181a attenuates H2O2-induced oxidative stress and cellular senescence in HFF by targeting PDIA6.

Clinical analysis of chronic mild occupational carbon disulfide poisoning / 中国职业医学

Objective: To analyze clinical features and electroneuromyography (ENMG) results of chronic mild occupational carbon disulfide poisoning cases. Methods: A total of 344 patients diagnosed with chronic mild occupational carbon disulfide poisoning based on GBZ 4-2002 Diagnostic Criteria of Occupational Chronic Carbon Disulfide Poisoning were selected as study subjects from 2006 to 2019 using the retrospective study method. Their clinical data was collected and analyzed. Results: The main symptoms of the study subjects were dizziness, headache, insomnia, dreaming, memory impairment, numbness and weakness in the distal extremities. Positive signs mainly included symmetrical glove and stocking distribution like sensory disorders in the distal extremities, and the weakening or absent Achilles tendon reflex and knee reflex. The incidence of symptoms and signs increased with the length of service (all P<0.01). The incidence of fundus and venous changes in patients was 41.3%, which increased with the length of service (P<0.01). ENMG examination showed varying degrees of abnormalities in the peripheral motor and/or sensory nerves in all patients, with a higher incidence of motor nerve abnormalities than sensory nerve abnormalities (21.1% vs 3.7%, P<0.01). The incidence of motor nerve abnormality was higher on the right side than the left side (23.7% vs 18.5%, P<0.01). The incidences of motor nerve abnormalities from high to low in the order were median nerve, common peroneal nerve, ulnar nerve and posterior tibial nerve (34.9% vs 27.9% vs 16.6% vs 5.1%, P<0.01). The incidences of sensory nerve abnormalities from high to low in the order were median nerve, ulnar nerve and sural nerve (5.2% vs 5.1% vs 0.7%, P<0.01). The incidences of left ulnar nerve, right ulnar nerve and right median nerve were higher in male patients than in female patients (15.2% vs 5.3%, 24.0% vs 11.7%, 44.8% vs 28.7%, all P<0.05), while the incidences of the left and right common peroneal nerve in lower extremity motor nerve were lower in male patients than in female patients (18.4% vs 52.1%, 21.2% vs 46.8%, all P<0.01). Conclusion: Chronic mild occupational carbon disulfide poisoning was mainly manifested as multiple peripheral nerve injury. ENMG results showed that the distal motor nerve conduction abnormalities were more sensitive than the sensory nerve conduction abnormalities, with a higher degree of impairment in the upper limb than the lower limb, and more impairment in the right side than the left side.

miRNA-4686 effects the proliferation and migration of esophageal cancer cells via targeting PDIA4 / 肿瘤研究与临床

Objective:To investigate the expressions of miRNA-4686 (miR-4686) and protein disulfide isomerase A4 (PDIA4) in esophageal cancer tissues, and the effect of miR-4686 on the proliferation and migration of esophageal cancer cells in vitro and possible mechanisms.Methods:The complementary integration of miR-4686 and PDIA4 mRNA were predicted using the miRWalk version 3 online website. The expression profiles of miR-4686 and PDIA4 mRNA were obtained from the University of California, Santa Cruz Genome (UCSC) database, and the relative expression of miR-4686 and PDIA4 mRNA in esophageal cancer tissues and paracancerous tissues were analyzed. The UCSC database was used to obtain the localization of miR-4686 in human cells. The relative expressions of miR-4686 and PDIA4 mRNA in esophageal cancer Eca109, TE-13, EC9706 and KYSE-510 cells and normal esophageal epithelial HET-1A cells were detected by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). Eca109 cells with the lowest relative expression of miR-4686 were selected for subsequent research, and the Eca109 cells were divided into miR-4686 group (transfected with miR-4686 mimic) and miR-NC group (transfected with miR-4686 negative control sequence mimic). The proliferation ability of Eca109 cells in the two groups was detected by colony formation assay, the migration ability of Eca109 cells was detected by scratch healing assay, and the targeting relationship between miR-4686 and PDIA4 mRNA was verified by dual luciferase reporter gene assay; the expressions of PDIA4 protein and PI3K-AKT-mTOR signaling pathway-related proteins were detected by Western blotting.Results:Compared with the paracancerous tissues, miR-4686 expression was low and PDIA4 mRNA expression was high in esophageal cancer tissues (both P < 0.01). Compared with normal esophageal epithelial HET-1A cells (0.98±0.15), the relative expression of miR-4686 in esophageal cancer Eca109 (0.11±0.04), TE-13 (0.58±0.10), EC9706 (0.34±0.05) and KYSE-510 cells (0.69±0.06) were all decreased (all P < 0.05). Compared with the miR-NC group, the relative expression of miR-4686 in Eca109 cells of the miR-4686 group increased (9.4±2.1 vs. 1.0±0.4, t = 3.88, P = 0.008), the number of colony formation decreased (38±9 vs. 114±18, t = 3.78, P = 0.009), the scratch healing rate decreased [(27.13±0.91)% vs. (45.05±3.89)%, t = 4.48, P = 0.004], and the relative expression of PDIA4 mRNA decreased [1.0±0.5 vs. 6.3±0.9, t = 5.04, P = 0.002]. Compared with wild type (WT)-PDIA4+miR-NC group, the relative luciferase activity of Eca109 cells in WT-PDIA4+ miR-4686 group decreased (0.31±0.08 vs. 0.99±0.08, t = 5.96, P < 0.001). Compared with miR-NC group, the expressions of PDIA4, p-PI3K, p-AKT and p-m-TOR proteins in Eca109 cells of the miR-4686 group were all decreased. Conclusions:In esophageal cancer tissues, miR-4686 is lowly expressed and PDIA4 mRNA is highly expressed. miR-4686 may inhibit the proliferation and migration of esophageal cancer Eca109 cells by targeting regulation of PDIA4 expression.

Co-load of silybin and doxorubicin by MoS2 nanosheets for synergetic chemotherapy and photothermal therapy of lung cancer / 药学学报

The active ingredient of traditional Chinese medicine, silybin (SBN), can inhibit the proliferation of cancer cells and enhance the anticancer effect of doxorubicin (DOX). However, due to non-targeting and short half-life of SBN and DOX, as well as different administration routes and pharmacokinetic processes, this combination drug cannot act on the tumor in the set order, seriously eliminating the synergistic effect between them and limiting the effect in vivo. Therefore, we intended to construct a nano-delivery system based on molybdenum disulfide (MoS2), modified by polyethylene glycol (PEG) and sialic acid (SA), and co-loaded with SBN and DOX. The system induced the release of combined drugs under the dual-stimulation of pH and near infra-red (NIR), increased the free concentration of intracellular drugs, so as to achieve the synergistic effect between them. The animal welfare and experimental procedures were in accordance with the regulations of the Animal Ethics Committee of Fujian University of Traditional Chinese Medicine. MoS2-PEG-SA-SBN/DOX circulated in vivo, and effectively accumulated at tumor sites through enhanced permeability and retention effect (EPR) and SA-mediated active targeting. Under near infrared light irradiation, MoS2-PEG-SA-SBN/DOX realized the combination of synergistic chemotherapy and photothermal therapy for tumor, thus achieving excellent anti-tumor effect in vivo. This study can provide a new idea and strategy for the clinical treatment of lung cancer. Taken together, MoS2-PEG-SA-SBN/DOX can offer a new idea and strategy for the clinical treatment of lung cancer.

Expression, thermal stability modification and application in PHB degradation of polyhydroxyalkanoate depolymerase from Thermomonospora umbrina / 生物工程学报

Polyhydroxyalkanoate depolymerase (PHAD) can be used for the degradation and recovery of polyhydroxyalkanoate (PHA). In order to develop a PHAD with good stability under high temperature, PHAD from Thermomonospora umbrina (TumPHAD) was heterelogously expressed in Escherichia coli BL21(DE3). At the same time, a mutant A190C/V240C with enhanced stability was obtained via rational design of disulfide bonds. Characterization of enzymatic properties showed that the mutant A190C/V240C had an optimum temperature of 60 ℃, which was 20 ℃ higher than that of the wild type. The half-life at 50 ℃ was 7 hours, at 50 ℃ which was 21 times longer than that of the wild type. The mutant A190C/V240C was used for the degradation of polyhydroxybutyrate (PHB), one of the typical PHA. At 50 ℃, the degradation rate of PHB being treated for 2 hours and 12 hours was 2.1 times and 3.8 times higher than that of the wild type, respectively. The TumPHAD mutant A190C/V240C obtained in this study shows tolerance to high temperature resistance, good thermal stability and strong PHB degradation ability, which may facilitate the degradation and recovery of PHB.

Inhibition of liver glutaminase activity by Diallyl disulfide in experimentally induced hepatoma in mice

Background: Cancer cells addiction to glutamine, an essential non-essential amino acid, has stirred up the interest in researchers across the globe. Increased glutamine metabolism (glutaminolysis) is a hallmark of cancer. Targeting glutaminolysis via glutaminase inhibition emerges as a promising strategy to disrupt cancer metabolism and tumor progression. Diallyl disulfide (DADS), a major organosulfur compound derived from garlic, is known for its anticancer properties. The mechanisms of action of DADS include activation of metabolic enzymes that detoxify carcinogens, suppression of the formation of DNA adducts, antioxidant effects, regulation of cell-cycle progression, induction of apoptosis, and inhibition of angiogenesis and metastasis. Aim: to assess the effect of diallyl disulfide on liver glutaminase activity in experimentally induced hepatoma in mice. Materials & Methods: Swiss albino male mice were divided into four groups - normal, control, preventive and curative groups. Hepatoma was induced by intraperitoneal injection of Ehrlich ascites carcinoma (EAC) cells. DADS (100 mg/kg body weight/mouse/day) was orally fed to protective and curative group mice for a stipulated time period. Mice of all the groups were sacrificed, and liver tissue glutaminase activity were measured. Results: The present study shows a significant decrease in glutaminase activity in protective (p >0.001) and curative groups (p >0.001) as compared to control group. Conclusion: DADS at the dosage employed shows inhibitory effects on liver glutaminase activity which may be attributed to anti-inflammatory properties of DADS, specifically in suppression of NF-kB signalling pathway.

Correspondência de teor de dissulfeto de captopril com odor de enxofre em comprimidos de captopril 25 mg, provenientes de farmácias públicas (postos de saúde) e drogarias comerciais / Correspondence of captopril disulfide content with sulfur odor in 25 mg captopril tablets from public pharmacies (health clinics) and commercial drugstores

Introdução: O captopril (CP) é o medicamento de escolha para o tratamento da hipertensão arterial. Sua degradação leva à formação do dímero dissulfeto de captopril (DSCP), este associado a um odor forte no medicamento, podendo causar abandono do tratamento pelo paciente. Objetivo: Determinar DCSP, associar a percepção olfativa de odor de enxofre desprendido do produto e realizar a avaliação de bula de comprimidos de captopril 25 mg distribuídos pelos setores público e privado. Método: Foi verificado o desempenho do método de determinação do CP e DSCP pela Farmacopeia Brasileira 6a ed. por HPLC (DAD). Foram analisados 13 produtos de comprimidos de captopril 25 mg, sendo dois provenientes do setor público de lotes diferentes e mesmo fabricante e 11 do setor privado de diferentes fabricantes e lotes. Foram avaliados aspectos do comprimido quanto à percepção de odor, determinação de peso, identificação e teor de CP e de DSCP e análise do conteúdo da bula. Resultados: Dentre os 13, o medicamento vencido apresentou 4,4% de DSCP, os demais estavam de acordo com a especificação. Verificouse  correspondência do odor de enxofre perceptível com teor de DSCP acima de 0,5%. Considerando os textos de bula sobre odor de enxofre, as constatações foram: nenhuma informação (três produtos), odor característico (dois), leve odor de enxofre (um), leve odor de enxofre sem diminuir a eficácia (sete). Conclusões: As amostras apresentaram resultados satisfatórios para os ensaios realizados. Verificou-se falta de homogeneidade nas informações das bulas sobre o odor dos comprimidos. A percepção do paciente quanto ao odor de enxofre, mesmo dentro do limite tolerado de DSCP, pode levar a não aceitação do medicamento e consequente não adesão ao tratamento da hipertensão, além de gerar prejuízos ao SUS.

Introduction: Captopril (CP) is the drug of choice for the treatment of hypertension. Its degradation leads to the formation of captopril disulfide dimer (DSCP), associated with a strong odor in the drug, which can cause the patient abandonment of treatment. Objective: To determine DCSP, associate the olfactory perception of the sulfur odor given off by the product and carry out the evaluation of the package insert for captopril 25 mg tablets distributed in the public and private sectors. Method: The performance of CP and DSCP determination method of the Brazilian Pharmacopoeia 6 ed was verified by HPLC (DAD). Thirteen products of 25 mg captopril tablets were analyzed, 2 of which came from the public sector from different batches and the same manufacturer: the other 11 came from the private sector from different batches and manufacturers. The samples were analyzed regarding appearance, odor perception, identification, weight determination, CP and DSCP content (by HPLC) and package insert content. Results: Among the 13, the expired drug had 4.4% DSCP; the others were in accordance with the specification. Correspondence of perceptible sulfur odor was established for drugs with DSCP content above 0.5%. Considering the texts on sulfur odor in the package inserts, the findings were: none information (3 products), characteristic odor (2), slight sulfur odor (1), slight sulfur odor without decreasing effectiveness (7). Conclusions: The samples showed satisfactory results for the tests performed. There was a lack of homogeneity in the information in the package inserts about odor of the tablets. The patient's perception of sulfur odor, even within the tolerated limit of DSCP, can lead to non-acceptance of the drug and consequent non- adherence to the treatment of hypertension, in addition to causing damage to the SUS.

Effect of diallyl disulfide on liver nucleotide metabolism in experimentally induced hepatoma in mice.

Background: The antitumorigenic effects of active ingredient of garlic, diallyl disulfide (DADS), has been extensively studied & found to retard the growth of neoplastic cells than any other allyl sulfur compounds of garlic. Earlier we have reported antitomorogenic properties of DADS, showing tumor regression by interfering with the liver glucose utilization, protein synthesis as well as lipid synthesis in tumor cells. Aim: To assess the effect of diallyl disulfide on liver nucleotide metabolism in experimentally induced hepatoma in mice. Materials & Methods: Swiss albino male mice were divided into four groups - normal, control, preventive and curative groups. Hepatoma was induced by intraperitoneal injection of Ehrlich ascites carcinoma (EAC) cells. DADS (100 mg/kg body weight/mouse/day) was orally fed to protective and curative group mice for a stipulated time period. Mice of all the groups were sacrificed, and liver tissue adenosine deaminase (ADA) activity and uric acid (UA) levels were measured. Results: The present study shows a significant decrease in ADA activity and UA levels in protective (p >0.001) and curative groups (p >0.01) as compared to control group. Conclusion: DADS has inhibitory effects on nucleotide metabolism by inhibiting the activities of ADA and xanthine oxidase enzymes, and by reducing the production of deoxy ribonucleotides, probably by involving in thiol-disulfide exchange reactions.

Proptosis is associated with thiol-disulfide in patients with Graves' ophthalmopathy

ABSTRACT Objective: Graves' ophthalmopathy (GO) is a vision-threatening finding observed in approximately half of Graves' disease patients. The pathophysiology of GO is unclear, and one of the suspected factors is oxidative stress. In our study, we compared the relationship between proptosis and SH-SS in patients diagnosed with GO. Materials and methods: In this prospective study, 40 recently diagnosed Graves' disease patients with proptosis, 40 recently diagnosed Graves' disease patients without GO and 30 healthy individuals with similar demographic characteristics were included. Serum thiol-disulfide (SH-SS) measurements were performed. Eye examinations were performed by a single ophthalmologist to check for the presence of GO, and proptosis values were recorded with a Hertel exophthalmometer. Results: Total SH values were lower in the group with proptosis than in the other groups (p < 0.05). Total and native SH values were lower in patients without proptosis than in the control group (p < 0.05). Total SH, native SH and SS levels were independently associated with proptosis (p < 0.05). According to this analysis, it was found that increasing SS and decreasing total and native SH levels increased the probability of proptosis by 24.4%, 32.7% and 32.4%, respectively. Conclusion: A decrease in SH, which is a natural antioxidant that protects the body against oxidative stress, and an increase in SS are important signs of oxidative damage. Proptosis and SH-SS are closely related in GO. This may help us detect GO and proptosis in Graves' patients. It can also assist in developing new options for preventing and treating GO.

Are thiol-disulfide homeostasis & neutrophil-to-lymphocyte ratio useful in the differential diagnosis of acute familial Mediterranean fever attacks & acute appendicitis?

Background & objectives: Continuous subclinical inflammation can be seen in patients with familial Mediterranean fever (FMF) during the attack-free period. The importance of oxidative stress parameters in acute appendicitis (AA) progression has also been shown in previous studies. So, oxidative stress and the oxidant/antioxidant balance may play a role in this persistent subclinical inflammation. With this background the main objective of this study was to investigate the usefulness of combining the thiol-disulfide homeostasis parameters and the neutrophil-to-lymphocyte ratio (NLR) in the differential diagnosis of AA and an acute FMF attack. Methods: The present study was conducted prospectively with 84 patients who were admitted to the emergency department between May 1, and December 31, 2018. Another 40 healthy individuals were assigned as the control group. The homeostasis parameters of thiol-disulfide were measured by a spectrophotometric method and NLR was measured in the patient and control groups. Results: Native thiol and total thiol values were lower, while disulfide values were insignificantly higher in patients with AA than in patients with FMF. The white blood cell (WBC), neutrophil and NLR values were significantly higher in the AA group (P<0.001, P<0.001, P<0.001, respectively). When the neutrophil cut-off value for AA was set at 8.55, the calculated sensitivity was 80 per cent, the specificity was 72.2 per cent, and the area under the curve was 0.837. Interpretation & conclusions: The results of this study suggest that neutrophil, WBC and NLR values can be useful in the differentiation of AA from an acute FMF attack.

Diallyl disulfide and diallyl trisulfide in garlic as novel therapeutic agents to overcome drug resistance in breast cancer / 药物分析学报

Breast cancer is one of the leading causes of cancer-related deaths in women worldwide.It is a cancer that originates from the mammary ducts and involves mutations in multiple genes.Recently,the treatment of breast cancer has become increasingly challenging owing to the increase in tumor het-erogeneity and aggressiveness,which gives rise to therapeutic resistance.Epidemiological,population-based,and hospital-based case-control studies have demonstrated an association between high intake of certain Allium vegetables and a reduced risk in the development of breast cancer.Diallyl disulfide(DADS)and diallyl trisulfide(DATS)are the main allyl sulfur compounds present in garlic,and are known to exhibit anticancer activity as they interfere with breast cancer cell proliferation,tumor metastasis,and angiogenesis.The present review highlights multidrug resistance mechanisms and their signaling pathways in breast cancer.This review discusses the potential anticancer activities of DADS and DATS,with emphasis on drug resistance in triple-negative breast cancer(TNBC).Understanding the anticancer activities of DADS and DATS provides insights into their potential in targeting drug resistance mecha-nisms of TNBC,especially in clinical studies.

Clinical significance of PDIA3 expression in hepatocellular carcinoma and its effect on the expression of IL6 and IL17 / 中华内分泌外科杂志

Objective:To investigate the clinical significance of protein disulfide isomerase A3 (PDI) A3 (PDIA3) expression in hepatocellular carcinoma tissues and its effect of PDIA3 on the expression of IL6 and IL17 in hepatocellular carcinoma cells.Methods:Immunohistochemistry was used to detect the expression of PDIA3 in the tissues of 72 patients with liver cancer and their adjacent tissues. HepG2 cells were divided into experimental group and control group. The cells in the experimental group were transfected with PDIA3-siRNA plasmid, and the cells in the control group were transfected with MOCK-siRNA plasmid. Fluorescence quantitative PCR was used to detect the content of PDIA3 mRNA in each group of cells. The expressions of PDIA3, IL6 and IL17 in each group of cells were detected by Western blot. The proliferation ability of each group of cells was detected by CCK8.Results:The positive rate of PDIA3 in liver cancer tissues was 85.22% (75/88), and the expression rate in adjacent tissues was 6.81% (6/88). The expression rate of PDIA3 in liver cancer tissues was significantly higher than that in adjacent tissues. The difference was statistically significant ( P<0.001). After transfection of siRNA, the expression levels of PDIA3 mRNA in HepG2 cells in the experimental group and control group were 1.23±0.20 and 0.43±0.12, respectively, and the expression levels of PDIA3 protein were 1.19±0.11 and 0.23±0.08, respectively. The expression levels of IL6 were 1.11±0.15 and 0.57±0.09, respectively. The expression levels of IL17 were 1.19±0.14 and 0.45±0.08, respectively, and the expressions of IL6 and IL17 were significantly decreased (all P<0.05). The absorbance of HepG2 cells in the experimental group and the control group at 120 h was 2.28±0.10 and 1.11±0.09, respectively, and the cell proliferation ability of the experimental group was significantly decreased ( P<0.05) . Conclusions:The expression of PDIA3 is significantly increased in hepatocellular carcinoma, which may be related to the malignancy of hepatocellular carcinoma. PDIA3 affects the proliferation of hepatocellular carcinoma cells by regulating the expression of IL6 and IL17.

Determination of creatinine and 2-thiothiazolidine-4-carboxylic acid in urine by ultra high performance liquid chromatography tandem mass spectrometry / 中华劳动卫生职业病杂志

Objective: To establish an ultrahigh performance liquid chromatography tandem mass spectrometry method for the determination of creatinine (Cre) and 2-thiothiazolidine-4-carboxylic acid (TTCA) in urine. Methods: In October 2020, the end-of-shift urine samples of the monitored subjects were taken, and the filtrate was prepared by centrifugation. After separated by ultra high performance liquid chromatography C18 column, acetonitrile and 0.2% acetic acid aqueous solution were used as mobile phases for gradient elution, the three quadrupole tandem mass spectrometry adopted an electrospray ion source (ESI) , the ion source temperature was 500 ℃ , and the air curtain gas flow rate was 31.4 L/min, qualitative and quantitative analysis of Cre and TTCA were carried out under the multiple reaction monitoring mode. Results: The linear range of Cre was 1.0-1 000.0 μg/L, the linear equation was y=947.3x-1605.6, and the correlation coefficient was 0.9994. The detection limit and the limit of quantitation were 0.3, 1.0 μg/L. When the addition concentrations were 50.0, 150.0 and 450.0 μg/L, the recovery rates were 92.8%-94.6% , the intra assay precisions were 3.6%-5.7% , and the inter assay precisions were 3.4%-5.4%. The linear range of TTCA was 0.1-200.0 μg/L, the linear equation was y=1164.7x-2243.9, and the correlation coefficient was 0.9991. The detection limit and the limit of quantitation were 0.03, 0.1 μg/L. When the addition concentrations were 10.0, 40.0 and 160.0 μg/L, the recovery rates were 90.8%-93.6%, the intra assay precisions were 4.6%-7.4%, and the inter assay precisions were 4.4%-6.9%. Conclusion: The sample pretreatment process of the ultra high performance liquid chromatography tandem mass spectrometry method for the determination of Cre and TTCA in urine is simple, and the continuous determination of Cre and TTCA in urine can be realized only by switching mass spectrometry parameters under the same chromatographic conditions, which is accurate and efficient, and each performance index of the method meets the determination requirements.

Study on determination of 2-thioxothiazolidine-4-carboxylic Acid in urine by high performance liquid chromatography / 中华劳动卫生职业病杂志

Objective: To establish a high performance liquid chromatography method for the determination of 2-thioxothiazolidine-4-carboxylic acid (TTCA) in urine. Methods: After acidification with hydrochloric acid, TTCA in urine was first extracted by ethyl acetate with excessive sodium chloride, then gradient separated by a symmetry C18 column and then detected by a diode array detector. The quantification was based on a working curve of external standard method. Results: The linear relationship of TTCA in urine was good in the range of 0.03-10.00 mg/L, and the correlation coefficient was 0.9999. The detection limit and minimum quantitative concentration of TTCA in urine were 0.008 mg/L and 0.027 mg/L. The intra-assay precision of the method was 0.9%-1.4%, the inter-assay precision was 1.3%-3.5%, and the average recovery was 85.0%-92.7% while the concentrations of TTCA in urine was 0.8, 2.0 and 8.0 mg/L, respectively (n=6) . Conclusion: The gradient elution high performance liquid chromatography method has simple operation and high sensitivity, and it is suitable for the determination of TTCA on a low level in urine for occupational workers exposure to carbon disulfide.

Diallyl disulfide downregulates XIAP through miR-7 to inhibit proliferation, migration and invasion of human gastric cancer SGC7901 cells / 中国药理学通报

Aim To investigate whether diallyl disul¬fide ( DADS) downregulates X-linked inhibitor of ap- optosis protein ( XIAP) through miR-7 , inhibiting the proliferation, migration anrl invasion of gastric eaneer SGC7901 eel Is.Methods Gastric eaneer SGC7901 eell line overexpressing XIAP was established.qRT- PCR and Western blot were used to deteet the effeet of DADS on XIAP expression in overexpressed eells.CCK-8 and plate elone formation assay were used to analyze the effeet of DADS on the proliferation of XIAP overexpressing eells.Wound healing and Transwell ex¬periments were employed to analyze the effeets of DADS on the migration and invasion of XIAP overex¬pressing eells.qRT-PCR was used to deteet the effeet of DADS on the expression of miR-7.Negative regula¬ tion of miR-7 on XIAP was verified by using dual lucif- erase reporter gene assay, qRT-PCR ,anrl Western blot.Results XIAP overexpression enhanced the prolifera¬tion , migration and invasion of SGC7901 cells.DADS downregulated XIAP and inhibited the proliferation, mi¬gration and invasion of XIAP overexpressing cells.DADS upregulated miR-7 expression.miR-7 bound and regulated XIAP 3 'UTR directly.Overexpression of miR-7 decreased XIAP expression,while knockdown of miR-7 increased XIAP expression.Conclusion DADS downregulates XIAP through miR-7 to inhibit SGC7901 cell proliferation, migration and invasion.

Interdomain Disulfide Bond Enhances the Stability of Single-chain Antibody Against Pro-gastrin-releasing Peptide / 中国生物化学与分子生物学报

Pro-gastrin-releasing peptide (ProGRP) is a specific marker of small cell lung cancer.