CRISPR-Cas and catalytic hairpin assembly technology for target-initiated amplification detection of pancreatic cancer specific tsRNAs

Transfer RNA-derived small RNAs (tsRNAs) tRF-LeuCAG-002 (ts3011a RNA) is a novel class of non-coding RNAs biomarker for pancreatic cancer (PC). Reverse transcription polymerase chain reaction (RT-qPCR) has been unfit for community hospitals that are short of specialized equipment or laboratory setups. It has not been reported whether isothermal technology can be used for detection, because the tsRNAs have rich modifications and secondary structures compared with other non-coding RNAs. Herein, we have employed a catalytic hairpin assembly (CHA) circuit and clustered regularly interspaced short palindromic repeats (CRISPR) to develop an isothermal and target-initiated amplification method for detecting ts3011a RNA. In the proposed assay, the presence of target tsRNA triggers the CHA circuit that transforms new DNA duplexes to activate collateral cleavage activity of CRISPR-associated proteins (CRISPR-Cas) 12a, achieving cascade signal amplification. This method showed a low detection limit of 88 aM at 37 °C within 2 h. Moreover, it was demonstrated for the first time that, this method is less likely to produce aerosol contamination than RT-qPCR by simulating aerosol leakage experiments. This method has good consistency with RT-qPCR in the detection of serum samples and showed great potential for PC-specific tsRNAs point-of-care testing (POCT)

Control technique for linepack of natural gas pipeline network

The control technique for linepack of natural gas pipeline network has been studied to ensure the safe and stable operation of gas pipeline network. The classification, control principle and control standard of linepack were elaborated based on the current production and operation experience of natural gas pipeline network, and the control technique for linepack was analyzed in detail from four aspects: selling in spring and autumn, guaranteed supply in winter and summer, gas distribution and prediction of accident pipeline survival time. The results suggest that linepack should be actively controlled for a uniform change according to the weekly inhomogeneity sale in spring and autumn, and it should be raised to the emergencyhigh level or the highest level to ensure the gas supply in winter and summer. The gas from the natural gas pipeline network can be reasonably distributed based on the control area of the linepack, and the survival time of the accident pipeline can be predicted according to the low linepack limit of natural gas pipeline

Heparin-Binding Protein: A Novel Biomarker Linking Four Different Cardiovascular Diseases

Cardiovascular diseases are an important group of diseases that seriously affect quality of life. Thus, their treatment warrants further study. Heparin-binding protein (HBP) is a granulocyte protein derived from neutrophils. When an infection occurs, neutrophils release HBP, which can lead to elevated HBP levels in the blood. Therefore, HBP family members are said to be important indicators of infection. However, basic evidence is still lacking to confirm the possible relationship between HBP and cardiovascular diseases. Using bioinformatics methods, we investigated the role of the HBP network in normal hearts and hearts from patients with cardiovascular disease. First, we used the Open Targets database to obtain a list of HBP-encoding mRNAs related to atherosclerosis, myocarditis, myocardial infarction, and myocardial ischemia. Then, we constructed an HBP gene interaction network map using STRING. Clustering coefficients were calculated using Cytoscape, and MCODE was used for subnet analysis. Finally, the proposed interstitial network of HBPs was established and analyzed by Metascape enrichment analysis of the relevant signaling pathways. The aggregation coefficient of the HBP interaction network was higher among hearts with the four cardiovascular diseases, atherosclerosis (0.496), myocarditis (0.631), myocardial infarction (0.532), and myocardial ischemia (0.551), than in normal hearts. Metascape analysis showed that “NABA_MATRISOME_ASSOCIATED” was a typical pathway with the highest p value associated with epithelialization in all four diseases. Moreover, a large number of important HBPs were identified that may be significant for the treatment of these diseases. Therefore, HBPs do have a highly atopic connectivity network in cardiovascular diseases, and specific HBPs or signaling pathways may be used as targets for the development of new treatments for cardiovascular diseases

Efficacy and Prognostic Effect of Daratumumab-based Chemotherapy Regimen in Multiple Myeloma: a Real-world Study

Background Multiple myeloma (MM) is a common hematologic malignancy. With the emergence of a variety of new drugs, the survival rates of patients have increased, however, further discussion on how to achieve deeper and faster remission and reduce recurrence of patients is still needed. Objective To investigate the efficacy and safety of daratumumab in the chemotherapy of MM. Methods 73 MM patients treated with daletumab in Sichuan Provincial People's Hospital from January 2020 to July 2022 were included as study subjects. Baseline information of the included patients were collected through the electronic medical record system. All patients were treated with daletumab based regimen and followed up through outpatient and inpatient review and telephone once a month when it is not time for review, with the time of first use of darettuzumab since 2020-01-01 as the starting point, death, disease recurrence or end of follow-up as the ending point. Follow-up was up to 2022-07-31 with a median follow-up time of 6.5 (0.5, 26.5) months. Overall response rate (ORR) and complete response rate were used to evaluate the efficacy of the patients. Kaplan-Meier curves were plotted for different drug use, initial renal function, drug administration, cytogenetic risk stratification and M protein types, and compared by Log-rank test. Results The median age of the patients was 64 (30, 86) years, including 38 males and 35 females. 28 MM patients were treated with first-line darettuzumab, 13 were switched from other regiments to darettuzumab due to adverse reactions, and 32 patients had relapsed and refractory multiple myeloma (RRMM). 18 patients were lost during follow-up, and a total of 55 patients were available for efficacy assessment. After one course of treatment with the darettuzumab-based chemotherapy regimen, ORR was 72.7% with the complete response rate of 30.9%, including 3.6% (2/55) achieved strin-gent complete response (sCR) and 27.3% (15/55) achieved complete response (CR), 10.9% (6/55) achieved very good partial response (VGPR), 30.9% (17/55) achieved partial response (PR), 9.1% (5/55) achieved minimal response (MR), 12.7% (7/55) achieved stable disease (SD), and 5.5% (3/55) achieved disease progression (PD). The median progress free survival (PFS) of the patients was 6.0 (0.5, 26.5) months, and the median overall survival was 16 (3, 103) months. There was no statistically significant difference in the PFS survival curve between patients treated first-line with darettuzumab and RRMM patients (χ2=3.676, P=0.055). The difference of PFS survival curve between patients with normal and impaired renal function was statistically significant (χ2=12.51, P=0.004). There was a statistically significant difference in the survival curve of patients treated with daletumab + bortezomib + dexamethasone and those treated with daletumab + dexamethasone (χ2=6.691, P=0.009). There was no statistically significant difference in PFS survival curves between patients with non-high risk and high-risk cytogenetic classification (χ2=0.328, P=0.567). There was no statistically significant difference in PFS survival curves among patients with M protein types of IgA, IgG, IgD, light chain, double clone and non-secretory (P>0.05). The main adverse reactions of the patients were infusion reaction and myelosuppression, of which 21 patients had adverse infusion reactions at the first infusion, 21 patients had leukopenia, 40 patients had anemia and 16 patients had thrombocytopenia. Conclusion Darettuzumab has good curative effect in MM patients, which can be used as the first-line treatment for newly diagnosed patients, improve the clinical prognosis of RRMM patients, and prolong the survival time of patients

Comparison of the Indicators of Psychological Stress in the Population of Hubei Province and Non-Endemic Provinces in China During Two Weeks During the Coronavirus Disease 2019 (COVID-19) Outbreak in February 2020

BACKGROUND: During February 2020, the coronavirus disease 2019 (COVID-19) epidemic in Hubei Province, China, was at its height, requiring isolation of the population. This study aimed to compare the emotional state, somatic responses, sleep quality, and behavior of people in Hubei Province with non-endemic provinces in China during two weeks in February 2020. MATERIAL/METHODS: Questionnaires were completed by 939 individuals (357 men; 582 women), including 33 from Hubei and 906 from non-endemic provinces. The Stress Response Questionnaire (SRQ) determined the emotional state, somatic responses, and behavior. The Pittsburgh Sleep Quality Index (PSQI) was used to measure the duration of sleep and sleep quality. RESULTS: There were 939 study participants, aged 18–24 years (35.89%) and 25–39 years (35.57%); 65.92% were university students. During a two week period in February 2020, the emotional state and behavior of participants in Hubei improved, but the quality of sleep did not. Health workers and business people became increasingly anxious, but other professionals became less anxious. The data showed that most people in Hubei Province developed a more positive attitude regarding their risk of infection and the chances of surviving the COVID-19 epidemic. CONCLUSIONS: During a two-week period, front-line health workers and people in Hubei Province became less anxious about the COVID-19 epidemic, but sleep quality did not improve. Despite public awareness, levels of anxiety exist that affect the quality of life during epidemics, including periods of population quarantine. Therefore, health education should be combined with psychological counseling for vulnerable individuals

Correlation between Bcl-2 Expression and the Efficacy of Bcl-2 Inhibitors in Patients with Myelodysplastic Syndromes

Background The heterogeneity of myelodysplastic syndromes (MDS) is strong, with poor efficacy of existing therapeutic regimens and large individual differences in patient prognosis. B-cell lymphomato-2 (Bcl-2) protein is highly expressed in patients with myeloid tumors, which has been proved by previous to be associated with disease progression, chemotherapy resistance, and shortened overall survival. The Bcl-2 inhibitor Veneckla (VEN) is approved for elderly patients with newly diagnosed acute myeloid leukemia (AML) who are not eligible for intensive therapy, and there is less data on its use in MDS patients. The difference in Bcl-2 expression in MDS patients and its correlation with the efficacy and prognosis of VEN therapy haven't been reported. Objective To analyse the expression of Bcl-2 protein in MDS patients and assess its correlation with the efficacy and prognosis of VEN treatment. Methods The clinical data of 71 patients with MDS admitted to Sichuan Academy of Medical Sciences·Sichuan Provincial People's Hospital from July 2018 to December 2022 were retrospectively analyzed. Baseline data of patients including gender, age, blood routine, blood biochemistry, bone marrow hemocytology, flow cytometry, chromosome karyotype, myeloid gene mutation, fusion gene mutation and MDS-EB typing were collected. The expression of Bcl-2 protein was detected by immunohistochemical staining. According to the modified International Prognostic Score System (IPSS-R), the patients were divided into 5 risk levels of very low risk (0 case), low risk (1 case), medium risk (7 cases), high risk (40 cases) and very high risk (23 cases). Patients with bone marrow Bcl-2 positivity≥10% were defined as Bcl-2 positive and<10% as Bcl-2 negative. Patients were divided into the positive group (n=38) and negative group (n=33). Of 63 patients with high-risk and very high-risk MDS, 38 were treated and evaluated for efficacy, including 28 patients who received VEN combined with azacitidine (VA combination regimen) and 10 patients who received hypomethylated drugs (HMA, including azacitidine or decitabine) as monotherapy. The patients were followed up until 2022-12-30. Kaplan-Meier curve was plotted to analyze the overall survival time (OS) of different patients, and Log-rank test was used for comparison. Univariate Cox regression model was used to analyze the influencing factors of patients' OS. Results The proportions of fusion gene mutation and nuclear myeloid phosphoprotein 1 (NPM1) gene mutation in the positive group were higher than those in the negative group, and the difference was statistically significant (P<0.05). Of the 10 patients who received HMA monotherapy, 1 was Bcl-2 negative and 9 were Bcl-2 positive, after 14 days of treatment, 4 of them (40.0%) achieved composite complete response (CR/mCR). Of the 28 patients receiving the VA combination regimen, 12 were Bcl-2 negative and 16 were Bcl-2 positive, and after 14 days of treatment, 20 (71.4%) patients receiving the VA combination regimen achieved CR/mCR. There was no significant difference in CR/mCR ratio between the two treatment regimens (P=0.087). The median overall survival time (mOS) for patients with VA combination therapy was 6.9 months, with a mOS of 2.9 months for Bcl-2 negative patients and 7.4 months for Bcl-2 positive patients. There was no significant difference in survival rate between Bcl-2 positive patients and Bcl-2 negative patients (χ2=2.075, P=0.150). The mOS of Bcl-2 positive patients receiving HMA monotherapy was 4.3 months, and there was no significant difference in the survival rate comparing with Bcl-2 positive patients receiving VA combination regimen (χ2=0.824, P=0.364). Univariate Cox regression model analysis showed that age, Bcl-2 protein expression, mutant gene number, chromosome karyotype, MDS-EB typing, IPSS-R risk, and efficacy were not the influencing factors for OS in patients treated with VA combination regimen (P>0.05) . Conclusion Bcl-2 protein contributes to MDS fusion gene mutation and myeloid gene mutation; Bcl-2 positive expression was not associated with the survival of MDS patients. There was no difference in survival rate between Bcl-2 positive and Bcl-2 negative patients who received the VA combination regimen

Influence of Polycyclic Aromatic Compounds and Oxidation States of Soot Organics on the Metabolome of Human-Lung Cells (A549): Implications for Vehicle Fuel Selection

Decades of research have established the toxicity of soot particles resulting from incomplete combustion. However, the unique chemical compounds responsible for adverse health effects have remained uncertain. This study utilized mass spectrometry to analyze the chemical composition of extracted soot organics at three oxidation states, aiming to establish quantitative relationships between potentially toxic chemicals and their impact on human alveolar basal epithelial cells (A549) through metabolomics-based evaluations. Targeted analysis using MS/MS indicated that particles with a medium oxidation state contained the highest total abundance of compounds, particularly oxygen-containing polycyclic aromatic hydrocarbons (OPAHs) composed of fused benzene rings and unsaturated carbonyls, which may cause oxidative stress, characterized by the upregulation of three specific metabolites. Further investigation focused on three specific OPAH standards: 1,4-naphthoquinone, 9-fluorenone, and anthranone. Pathway analysis indicated that exposure to these compounds affected transcriptional functions, the tricarboxylic acid cycle, cell proliferation, and the oxidative stress response. Biodiesel combustion emissions had higher concentrations of PAHs, OPAHs, and nitrogen-containing PAHs (NPAHs) compared with other fuels. Quinones and 9,10-anthraquinone were identified as the dominant compounds within the OPAH category. This knowledge enhances our understanding of the compounds contributing to adverse health effects observed in epidemiological studies and highlights the role of aerosol composition in toxicity.</p