LncRNA NR_003508 Suppresses Mycobacterium tuberculosis-Induced Programmed Necrosis via Sponging miR-346-3p to Regulate RIPK1.

Emerging evidence suggests that long non-coding RNAs (LncRNAs) are involved in Mtb-induced programmed necrosis. Among these LncRNAs, LncRNA NR_003508 is associated with LPS-induced acute respiratory distress syndrome. However, whether LncRNA NR_003508 contributes to Mtb-induced programmed necrosis remains undocumented. Firstly, the expression of LncRNA NR_003508 was determined using RT-qPCR and FISH. The protein expression of RIPK1, p-RIPK1, RIPK3, p-RIPK3, MLKL, and p-MLKL was measured by Western blot in RAW264.7 and mouse lung tissues. Furthermore, luciferase reporter assays and bioinformatics were used to predict specific miRNA (miR-346-3p) and mRNA (RIPK1) regulated by LncRNA NR_003508. In addition, RT-qPCR was used to detect the RIPK1 expression in TB patients and healthy peripheral blood. The flow cytometry assay was performed to detect cell necrosis rates. Here we show that BCG infection-induced cell necrosis and increased LncRNA NR_003508 expression. si-NR_003508 inhibited BCG/H37Rv-induced programmed necrosis in vitro or in vivo. Functionally, LncRNA NR_003508 has been verified as a ceRNA for absorbing miR-346-3p, which targets RIPK1. Moreover, RIPK1 expression was elevated in the peripheral blood of TB patients compared with healthy people. Knockdown of LncRNA NR_003508 or miR-346-3p overexpression suppresses cell necrosis rate and ROS accumulation in RAW264.7 cells. In conclusion, LncRNA NR_003508 functions as a positive regulator of Mtb-induced programmed necrosis via sponging miR-346-3p to regulate RIPK1. Our findings may provide a promising therapeutic target for tuberculosis.

A HPLC-Q-TOF-MS-based urinary metabolomic approach to identification of potential biomarkers of metabolic syndrome.

Metabolic syndrome (MetS) is a serious threat to public health worldwide with an increased risk of developing type 2 diabetes, cardiovascular diseases and all-cause morbidity and mortality. In this study, a urinary metabolomic approach was performed on high performance liquid chromatography quadrupole time-of-flight mass spectrometry to discriminate 36 male MetS patients and 36 sex and age matched healthy controls. Pattern recognition analyses (principal component analysis and orthogonal projections to latent structures discriminate analysis) commonly demonstrated the difference between MetS patients and no-MetS subjects. This study found 8 metabolites that showed significant changes in patients with MetS, including branch-chain and aromatic amino acids (leucine, tyrosine, phenylalanine and tryptophan), short-chain acylcanitine (tiglylcarnitine), tricarboxylic acid (TCA) cycle intermediate (cis-aconitic acid) and glucuronidated products (cortolone-3-glucuronide and tetrahydroaldosterone-3-glucuronide). The candidate biomarkers revealed in this study could be useful in providing clues for further research focusing on the in-depth investigation of the cause of and cure for MetS.

Integration of protein L-immobilized epoxy magnetic bead capture with LC-MS/MS for therapeutic monoclonal antibody quantification in serum.

In recent years, there has been a growing interest in the thriving monoclonal antibody (mAb) industry due to the wide utilization of mAbs in clinical therapies. Robust and accurate bioanalytical methods are required to enable fast quantification of mAbs in biological matrices, especially in the context of pharmacokinetics (PKs)/pharmacodynamics (PDs) and therapeutic drug monitoring (TDM) studies. In this investigation, we presented a novel immuno-magnetic capture coupled with a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method designed for the quantification of immunoglobulin G-kappa-based mAbs in biological fluids. The immunoaffinity absorbent for mAb drug purification was meticulously crafted by immobilizing protein L onto monosize, magnetic poly(glycidyl methacrylate) (m-pGMA) beads, synthesized through dispersion polymerization. The microspheres were acquired with an average size of 1.6 µm, and the optimal binding of mAbs from the aqueous mAb solution was determined to be 45.82 mg g-1. The quantification of mAbs in 10 µL serum samples was achieved through affinity purification using m-pGMA@protein L beads (employing rituximab as an internal standard (IS)), on-bead reduction, and rapid tryptic digestion. Remarkably, the entire process, taking less than 2.5 hours, held significant potential for simplifying pretreatment procedures and minimizing analytical time. Furthermore, the developed method underwent validation in accordance with the European Medicines Agency (EMA) guidelines. The assay demonstrated commendable linearity within the 2-400 µg mL-1 range for both daratumumab and pembrolizumab. Intra- and inter-assay coefficients of variation fell within the range of 0.7% to 13.4%, meeting established acceptance criteria. Other validation parameters also conformed to regulatory standards. Ultimately, the efficacy of the method was substantiated in a pharmacokinetic study following a single-dose intravenous administration to mice, underscoring its applicability and reliability in real-world scenarios.

A case report: An unusual presentation of adenoid cystic carcinoma of the lacrimal gland.

RATIONALE: Adenoid cystic carcinoma (ACC) is the most common malignant epithelial tumor of the lacrimal gland with the highest malignant degree. ACC of the lacrimal gland is characterized by symptoms of <1 years duration. We present a 38-year-old male patient who complained of an enlarging mass in the left lacrimal fossa for almost 10 years previous to the diagnosis of ACC. PATIENT CONCERNS: A 38-year-old male patient visited our ophthalmology clinic with a chief complaint of a mass in his left upper lid, which had enlarged significantly over the previous months. DIAGNOSES: Magnetic resonance imaging with intravenous Gadobutrol showed moderate and homogenous mass enhancement. Bone destruction is found. The periosteum is not eroded. The magnetic resonance imaging finding was supportive for malignancy. Histopathological examination of the specimen revealed solid tumor showing a cribriform pattern mixed small amount of basaloid cell proliferation. Therefore, the final diagnose was Adenoid cystic carcinoma of the lacrimal gland. INTERVENTIONS: The treatment included en bloc resection of the mass and adjacent bone and radiotherapy. OUTCOMES: In 1 year follow-up after operation, there is no recurrence. Visual acuity is 30/30. The left eye shows limitation on abduction. LESSONS: The present case demonstrates an unusual progression of ACC of the Lacrimal Gland.

3,4,5-Trimeth-oxy-phenol.

The asymmetric unit of the title compound, C(9)H(12)O(4), consists of two crystallographically independent mol-ecules with similar conformations: essentially planar [r.m.s deviations for C(6)O(4) = 0.0057 and 0.0137 Å] except for the central meth-oxy-methyl group [C-C-O-C torsion angles = 83.3 (2) and 83.9 (2)°]. In the crystal, O-H⋯O hydrogen bonds link the mol-ecules, generating supra-molecular chains along the b axis.The three-dimensional crystal structure is stabilized by C-H⋯O and C-H⋯π inter-actions.

[The relationship between occupational stress and metabolic syndrome among policemen].

OBJECTIVE: To explore the occupational stress factors associated with the prevalence of metabolic syndrome (MS) among male policemen. METHODS: Using cluster sampling method, we selected four Public Security Bureau within the jurisdiction of the station now in some city. All the male police were included as research objects, and finally 1490 persons were selected, health and occupational stress inventory-revised (OSI-R) questionnaire were used for epidemiological surveys, and anthropometric examination and chemical indicators were also measured at the same time. The analysis methods were chi-square test and unconditional logistical regression. RESULTS: Among the 1490 of research objects, 1483 completed the questionnaire, and 1480 of the eligible questionnaires were available.237 cases were MS, and the prevalence rate was 16.0%(237/1480). The number of cases who were high, moderate and lack of occupational stress in MS group were 8, 39 and 23, that in non-MS were 14, 114 and 131, respectively. The odds of occupational stress with the highest and medium among policemen than who were lack were 4.82 (95%CI: 1.50 - 15.41) and 3.33 (95%CI: 1.62 - 6.79); the average score of role ambiguity, role insufficiency and responsibility in the group of MS were (38.76 ± 6.83), (25.74 ± 7.22), (25.76 ± 6.27); and that in non-MS were (37.55 ± 6.85), (24.50 ± 6.58), (25.05 ± 5.95). The logistical regression analysis showed that: the likely three occupational risk stress factors which influencing the prevalence of MS were role ambiguity, role insufficiency and responsibility, and the OR (95%CI) were 1.06 (1.02 - 1.10), 1.04 (1.02 - 1.07) and 1.03 (1.01 - 1.06), respectively. CONCLUSION: Role ambiguity, role insufficiency and responsibility were the occupational risk stress factors associated with the prevalence of MS among male policemen.

[A matched case-control study on the risk factors of metabolic syndrome among policemen].

OBJECTIVE: To investigate the life style, genetic and occupational risk factors of metabolic syndrome (MS) among policemen. METHODS: 1:4 matched case-control study was used, based on physical examination data of Tianjin Policemen in 2010, 708 patients with MS were randomly selected as cases, which were matched with 2832 healthy controls on the basis of sex and age (+/- 1 year). An epidemiological investigations on the past exposure status of several possible risk factors was conducted, and the data were analyzed with conditional logistic regression. RESULTS: Fifteen factors related to exposure were identified for MS through univariate conditional logistic regression analysis. Multivariate conditional logistic regression analysis suggested that, seven factors, such as family history of hypertension (OR = 2.406, 95% CI: 1.946-2.975), family history of diabetes (OR = 1.301, 95% CI: 1.043-1.623), smoking (OR = 1.357, 95%CI: 1.010-1.823), snoring (OR = 1.268, 95% CI: 1.043-1.543), work intensity (OR = 4.603, 95% CI: 3.767-5.623), occupational stressful events (OR = 1.524, 95% CI: 1.209-1.922), security policemen (OR = 1.453, 95% CI: 1.127-1.872) and criminal investigation policemen (OR = 2.792, 95% CI: 2.168-3.596), could significantly increase the risk of disease development, but dairy products (OR = 0.782, 95% CI: 0.619-0.989) was a protect factor for MS. The results from population attributable risk factors analysis showed that the control of smoking, snoring, work intensity, occupational stressful events can decreased the risk of MS to 16.26%, 11.71%, 56.87% and 8.97%, respectively. CONCLUSION: Metabolic syndrome has became a significant public health problem among policemen, it's necessary to take measures on life style, occupational risk factors for reducing the incidence of MS, and improving the health level among policemen.

[The inhibitory effect of recombinant polypeptide CH50 of fibronectin on invasion and angiogenesis of tumors].

OBJECTIVE: To investigate the inhibitory effect of recombinant polypeptide CH50 of fibronectin on invasion and angiogenesis of tumors, and analyze the possible molecular mechanism of the therapeutic effect of polypeptide CH50 on tumors. METHODS: The tumor model was established by inoculation of H22 hepatocarcinoma cells in mice. The tumor gene therapy was performed by in vivo gene transfection with a method based on hydrodynamics to express polypeptide CH50. After treatment, the inhibitory effect on tumor invasion and angiogenesis was observed by histotology with HE staining of tumor tissues. The expresison of MMP-9 mRNA and protein at the edge of tumor tissue was evaluated by RT-PCR and gelatin zymography, respectively. RT-PCR was used to detect the expression of the related genes in H22 cells treated with polypeptide CH50. Cell adhesion assay was used to analyze the influence of polypeptide CH50 on the binding of cells to fibrinogen. RESULTS: (1) Eukaryotic expression plasmid pCH510 was expressed in vivo in a non-targeting manner and produced a significant inhibitory effect on tumor growth. The therapy with polypeptide CH50 resulted in pronounced necrosis of tumor cells in pCH510 group, compared with that in control groups at histological level. (2) Polypeptide CH50 could inhibit the growth, invasion and angiogenesis of the tumor, and interfere the formation of new collateral circulation in the tumor. (3) The expression level of MMP-9 protein at the edge of tumor tissue was significantly decreased after treatment, especially the activation of pro-MMP-9 was inhibited significantly, whereas the expression level of MMP-9 mRNA was not influenced. (4) The expression of alphav, 33 and cdc2 mRNAs in H22 cells treated with polypeptide CH50 was down-regulated. (5) Cell adhesion assay manifested that polypeptide CH50 can affect the adhesion ability of H22 cells. CONCLUSION: Polypeptide CH50 can inhibit tumor growth and angiogenesis by suppressing the functions of MMP-9 and integrin alphavbeta3.

Inhibition of hepatocarcinoma and tumor metastasis to liver by gene therapy with recombinant CBD-HepII polypeptide of fibronectin.

Unlike the intact fibronectin (FN) molecule, some proteolytic or recombinant fragments of FN possess inhibitory activities on tumor, providing potential strategies in tumor therapeutics. Using the hydrodynamics-based gene delivery technique, we demonstrated that the treatment by in vivo expression of a recombinant CBD-HepII polypeptide of FN, designated as CH50, strongly inhibited the tumor growth, tumor invasion and angiogenesis. Such inhibitory effects of CH50 on tumor were partly ascribed to its influence on the activities of MMP-9 and alphavbeta3 integrin. The in vivo expressed CH50 decreased both the production and the activity of MMP-9 in tumor tissues. CH50 also down-regulated alphavbeta3 expression in tumor cells and endothelial cells in vitro. The decreased activity of alphavbeta3 integrin was proved by its reduced binding ability to fibrinogen and the down-regulation of cdc2 expression. The gene therapy with CH50 not only prolonged the survival of mice bearing hepatocarcinoma in the liver, but also suppressed the growth and invasive ability of tumor in spleen and its metastasis to liver. Taken together, these findings suggest a prospective utility of CH50 in the gene therapy of liver cancer.