Application of improved GalNAc conjugation in development of cost-effective siRNA therapies targeting cardiovascular diseases.

N-Acetylgalactosamine (GalNAc)-conjugated small interfering RNA (siRNA) therapies have received approval for treating both orphan and prevalent diseases. To improve in vivo efficacy and streamline the chemical synthesis process for efficient and cost-effective manufacturing, we conducted this study to identify better designs of GalNAc-siRNA conjugates for therapeutic development. Here, we present data on redesigned GalNAc-based ligands conjugated with siRNAs against angiopoietin-like 3 (ANGPTL3) and lipoprotein (a) (Lp(a)), two target molecules with the potential to address large unmet medical needs in atherosclerotic cardiovascular diseases. By attaching a novel pyran-derived scaffold to serial monovalent GalNAc units before solid-phase oligonucleotide synthesis, we achieved increased GalNAc-siRNA production efficiency with fewer synthesis steps compared to the standard triantennary GalNAc construct L96. The improved GalNAc-siRNA conjugates demonstrated equivalent or superior in vivo efficacy compared to triantennary GalNAc-conjugated siRNAs.

Role of Up-Regulated Transmembrane Channel-Like Protein 5 in Pancreatic Adenocarcinoma.

BACKGROUND: Pancreatic adenocarcinoma (PAAD) is a malignant tumor responsible for a heavy disease burden. Previously, only one pan-cancer study of Transmembrane channel-like protein 5 (TMC5) showed that TMC5 was highly expressed in PAAD, but the results lacked comprehensive verification, and the mechanism of TMC5 in PAAD was still unclear. METHODS: For exploring the expression and clinical value of TMC5 in PAAD better, we adopted a comprehensive evaluation method, using internal immunohistochemistry (IHC) data combined with microarray and RNA-sequencing data collected from public databases. The single cell RNA-sequencing (scRNA-seq) data were exploited to explore the TMC5 expression in cell populations and intercellular communication. The potential mechanism of TMC5 in PAAD was analyzed from the aspects of immune infiltration, transcriptional regulation, function and pathway enrichment. RESULTS: Our IHC data includes 148 PAAD samples and 19 non-PAAD samples, along with the available microarray and RNA-sequencing data (1166 PAAD samples, 704 non-PAAD samples). The comprehensive evaluation results showed that TMC5 was evidently up-regulated in PAAD (SMD = 1.17). Further analysis showed that TMC5 was over-expressed in cancerous epithelial cells. Furthermore, TMC5 was up-regulated in more advanced tumor T and N stages. Interestingly, we found that STAT3 as an immune marker of Th17 cells was not only positively correlated with TMC5 and up-regulated in PAAD tissues, but also the major predicted TMC5 transcription regulator. Moreover, STAT3 was involved in cancer pathway of PAAD. CONCLUSION: Up-regulated TMC5 indicates advanced tumor stage in PAAD patients, and its role in promoting PAAD development may be regulated by STAT3.

Active Enhancer Assessment by H3K27ac ChIP-seq Reveals Claudin-1 as a Biomarker for Radiation Resistance in Colorectal Cancer.

BACKGROUND: Colorectal cancer (CRC) represents the third most common malignant tumor in the worldwide. Radiotherapy is the common therapeutic treatment for CRC, but radiation resistance is often encountered. ChIP-seq of Histone H3K27 acetylation (H3K27ac) has revealed enhancers that play an important role in CRC. This study examined the relationship between an active CRC enhancer and claudin-1 (CLDN1), and its effect on CRC radiation resistance. METHODS: The target CRC genes of active enhancers were obtained from public H3K27ac ChIP-seq, and the genes highly expressed in radio-resistant CRC were screened and intersected with enhancer-driven genes. The clinical roles of CLDN1 in radiation resistance were examined using the t-test, standard mean deviation (SMD), summary receiver operating characteristic curve and Kaplan-Meier curves. The co-expressed genes of CLDN1 were calculated using Pearson Correlation analysis, and Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes and Gene Set Variation Analysis (GSVA) analyses were used to examine the molecular mechanisms of CLDN1. RESULTS: Total 13 703 CRC genes were regulated by enhancers using 58 H3K27ac ChIP-seq. Claudin-1 (CLDN1) was enhancer-driven and notably up-regulated in CRC tissues compared to non-CRC controls, with a SMD of 3.45 (95 CI % = .56-4.35). CLDN1 expression was increased in radiation-resistant CRC with a SMD of .42 (95% CI = .16-.68) and an area under the curve of .74 (95% CI = .70-.77). The cell cycle and immune macrophage levels were the most significant pathways associated with CLDN1. CONCLUSION: CLDN1 as an enhancer-regulated gene that can boost radiation resistance in patients with CRC.

[Assessment of Emergency Emission Reduction Effect During the Heavy Air Pollution Episodes in Beijing, Tianjin, Hebei, and Its Surrounding Area("2+26" Cities) from October to December 2019].

To evaluate the effect of emergency emission reduction measures during the heavy air pollution episodes in Beijing, Tianjin, Hebei, and its surrounding areas, a scenario simulation method was used. The concentrations of PM2.5, PM10, SO2, NO2, CO, and O3-8h, air quality index (AQI), characteristics of heavy air pollution, and climate and meteorological factors were analyzed using the observation data available from October to December 2019. The 24 h, 72 h, and 144 h prediction results of NAQPMS model were analyzed. The uncertainties of the assessment and model prediction were discussed. The results showed that the average PM2.5 concentration in Beijing, Tianjin, and its surrounding 26 cities ("2+26" cities) from October to December 2019 was 64 µg ·m-3, indicating a decrease of 10 µg ·m-3 as compared with that during the same period in 2018. There were 4 occurrences of regional heavy air pollution episodes, with the average PM2.5 concentration of 156 µg ·m-3 of affected cities. The value of evaluation on meteorological condition index of PM2.5 pollution (EMI) of "2+26" cities ranged from -15.6%-16.8%. The meteorological conditions of 12 cities, including Beijing, Tianjin, and Shijiazhuang, deteriorated as compared with those during the same period in 2018, and the changes ranged from 3.2%-16.8%. However, the emergency emission reduction measures effectively reduced the occurrence of regional heavy air pollution episodes, the peak concentration of PM2.5 was decreased significantly, and no severe regional pollution episode occurred. The daily PM2.5 concentrations reduced by 2% to 9% in Beijing, Shijiazhuang, Baoding, Tangshan, and other cities during a typical heavy air pollution period. The quarterly average concentrations of PM2.5 in the "2+26" cities reduced by 1 to 3 µg ·m-3. The regional emergency emission reduction measures have played an active role in protecting the health of the people and improving the quality of ambient air.

[Heavy Pollution Characteristics and Assessment of PM2.5 Predicted Model Results in Beijing-Tianjin-Hebei Region and Surrounding Areas During November 23 to December 4, 2018].

This study discusses the concentration characteristics of PM2.5 and PM10, as well as pollution meteorology in large-scale and long-term heavy pollution in the Beijing-Tianjin-Hebei region and its surrounding areas from November 23 to December 4, 2018, where the primary pollutants are comprised of PM2.5 and PM10. The monitoring results obtained from ground-based and vehicle-mounted lidars, as well as the HYSPLIT-4 backward trajectory combined with meteorological factors analysis are discussed. The accuracy and uncertainty of the air quality forecast model of NAQPMS, CMAQ, and CAMx during heavy air pollution were analyzed retrospectively. The results show that PM2.5 and sand dust in most cities in the south-central region contribute to severe pollution levels. The hourly peak concentrations of PM10 in Zhangjiakou, Beijing, Shijiazhuang, Handan, and Zhengzhou were 1589, 864, 794, 738, and 766 µg·m-3, respectively. The respective hourly peak concentrations of PM2.5 were 239, 319, 387, 321, and 380 µg·m-3. Ground static pressure field, high humidity, inversion, and other static and stable conditions, as well as sand dust transmitted from the northwest, were important pollution meteorological and weather factors. The monitoring data of ground-based lidar and vehicle-mounted lidar combined with the HYSPLIT-4 backward trajectory analysis showed that the air pollutant transmitted from the Southwest and Southeast during the heavy pollution period was primarily PM2.5. The air pollutant transmitted from the Northwest during the two sand dust processes. Moreover, the model of NAQPMS, CMAQ, and CAMx performed well in forecasting the heavy pollution process in the Beijing-Tianjin-Hebei region and its surrounding areas. However there are slight deviations for some individual cities, related to uncertainty in the meteorological model prediction, atmospheric chemical reaction mechanism, and pollution source list. Furthermore, the reduction in pollution source emissions caused by pollution emergency measures was also one of the main reasons for the overestimation.

[Concentration Characteristics and Assessment of Model-Predicted Results of PM2.5 in the Beijing-Tianjin-Hebei Region in Autumn and Winter].

This paper discusses the concentration characteristics of PM2.5, as well as its relationship with meteorological factors in autumn and winter (from September to the following February), from 2013 to 2018 in the Beijing-Tianjin-Hebei (BTH) region. The accuracy and uncertainty of the air quality forecast models NAQPMS(nested air quality prediction modeling system), CMAQ(community multiscale air quality modeling system), and CAMx (comprehensive air quality model with extensions) were analyzed based on the model-predicted and measured PM2.5 concentration in autumn and winter from 2015 to 2018. The accuracy of NAQPMS, CMAQ, and CAMx during typical heavy air pollution was also tested. Moreover, methods to improve the accuracy of the model forecast were discussed. The results showed that the mean concentrations of PM2.5 in the BTH region were 122, 98, 82, 99, and 65 µg·m-3 in the five autumn and winter periods, respectively. When the air quality index (AQI) exceeded 150 during each autumn and winter, it reached 229, 198, 210, 204, and 180 µg·m-3, respectively. There were 64 occurrences of heavy regional PM2.5 air pollution in autumn and winter from 2013 to 2018. The average duration was longest in the 2013 to 2014 period, and shortest in the 2017 to 2018 period. The peak concentration and average concentration of PM2.5 decreased year on year, except for the period from 2016 to 2017. In autumn and winter, PM2.5 concentration had a relatively close relationship with relative humidity, wind and sunshine duration, compared with a weak relationship with temperature and air pressure. Regional heavy air pollution always happened under the condition of low wind speed(less than 2 m·s-1),higher relative humidity(greater than 65%),and southwest and northeast wind direction. In addition, the heavy air pollution of PM2.5 in BTH in autumn and winter can be effectively forecasted by NAQPMS, CMAQ, and CAMx. The predicted and measured PM2.5 concentration showed a close relationship. The models performed well in forecasting Zhangjiakou, Chengde, and Qinhuangdao, but by contrast overestimated in Tangshan, Shijiazhuang, Baoding, Beijing, and Tianjin. The uncertainty of emission sources, measured and predicted meteorological data, and the atmospheric chemical reaction mechanism may be the main reasons for the overestimate.

Free triiodothyronine in relation to coronary severity at different ages: Gensini score assessment in 4206 euthyroid patients.

OBJECTIVE: To study whether free triiodothyronine (FT3) within normal range has effects on the presence and severity of coronary artery disease (CAD) in different gender and age groups. METHODS: A total of 4206 euthyroid patients were consecutively enrolled and divided into CAD group (n = 3306) and non-CAD group (n = 900). All patients underwent coronary angiography (CAG). Gensini score (GS) was used to determine the severity of coronary artery stenosis. Severe CAD was defined as GS > 32 and mild CAD was defined as GS ≤ 32. Logistic regression analysis and linear regression analysis were conducted to determine the association of FT3 with CAD in patients with different gender and ages. RESULTS: Concentration of FT3 was lower in patients with CAD than that in angiography-normal control group (P < 0.05). In addition, concentration of FT3 was lower in severe CAD than that in mild CAD. After adjusting for traditional cardiovascular risk factors and potential confounders, FT3 was negatively correlated with the presence of CAD, but not in the old patients (> 65 years old). Multivariable linear regression analysis showed that FT3 was negatively associated with GS in male and young patients with stable CAD, but not in the old patients. CONCLUSIONS: Low FT3 within normal range was negatively associated with the presence and severity of CAD in young patients, but not in the old ones. Further studies are needed to confirm our findings.

Is the ratio of apoB/apoA-1 the best predictor for the severity of coronary artery lesions in Chinese diabetics with stable angina pectoris? An assessment based on Gensini scores.

BACKGROUND: There is a paucity of data about the best lipid ratio predicting the severity of coronary artery disease (CAD) in patients with diabetes mellitus. We determined the relationship between five conventional lipid ratios and the extent of coronary artery lesions in Chinese Type 2 diabetics with stable angina pectoris (SAP). METHODS: A prospective cohort study within 373 type 2 diabetic patients diagnosed with stable CAD by coronary angiography was performed. All patients were classified into three groups according to the tertiles of Gensini scores (GS, low group < 8 points n = 143; intermediate group 8-28 points, n = 109; high group > 28 points, n = 121). Association between the ratios of apolipoprotein (apo) B and apoA-1, total cholesterol and high density lipoprotein cholesterol (TC/HDL-C), triglycerides and HDL-C (TG/HDL-C), low density lipoprotein cholesterol and HDL-C (LDL-C/HDL-C), Non-HDL-C/HDL-C and GS were evaluated using the receivers operating characteristic (ROC) curves and multivariate logistic regression models. RESULTS: The ratio of apoB/apoA-1, TC/HDL-C, LDL-C/HDL-C, and Non-HDL-C/HDL-C were correlated with Gensini scores. Area under the ROC curves for predicting high Gensini scores in the ratios of apoB/apoA-1, TC/HDL-C, LDL-C/HDL-C and Non-HDL-C/HDL-C were 0.62, 0.60, 0.59 and 0.60, respectively (P < 0.005 for all). According to multivariate logistic regression analysis after adjusted with demographic characteristic and other lipid parameters, the ratio of apoB/apoA-1 is qualified as an independent discriminator for the severity of CAD. However, after further adjusting different baseline variables, such as left ventricular ejective fraction, hemoglobin A1c, leukocytes count and serum creatinine, none of the above lipid ratios remained. CONCLUSIONS: Compared with other lipid parameters, the ratio of apoB/apoA-1 appears to be more significantly correlated with the extent of coronary artery lesions in Chinese diabetics, but it was not an independent predictor in these settings.

High-density lipoprotein subfractions in relation with the severity of coronary artery disease: A Gensini score assessment.

BACKGROUND: The exact role of different high-density lipoprotein (HDL) subfractions in the pathogenesis of coronary artery disease (CAD) has not yet been fully explored. The aim of the present study was to examine the relationship between HDL subfractions and the severity of CAD in patients without statin therapy. METHODS: A total of 382 consecutive patients (mean: 55.36 ± 11.30 years of age) who underwent coronary angiography from angina-like chest pain were investigated. Patients were classified into 2 groups according to the angiographic results: a CAD group (n = 283) and a control group (n = 99). The distribution of HDL subfractions was analyzed using a Quantimetrix Lipoprint HDL system. CAD severity was measured by Gensini score (GS). RESULTS: Compared with the control group, HDL-cholesterol (HDL-C), large HDL-C level, and the large HDL subfraction percentages in the CAD group were significantly lower (P = .002, P < .001, P < .001, respectively). Meanwhile, a small HDL-C level and the percentage of small HDL subfraction were significantly higher (P = .003, P < .001, respectively). Correlation analysis showed that the percentage of a large HDL subfraction was negatively correlated with GS (ß = -0.191, P = .005), whereas the percentage of a small HDL subfraction positively correlated with GS (ß = 0.145, P = .023) in patients with CAD. CONCLUSIONS: Small HDL subfraction was associated with the presence of CAD, whereas the percentage of large HDL and small HDL subfraction was negatively and positively associated with the severity of CAD, respectively.

Optimal cutoff scores for dementia and mild cognitive impairment of the Montreal Cognitive Assessment among elderly and oldest-old Chinese population.

BACKGROUND: All versions of the Montreal Cognitive Assessment (MoCA) lack population-based data of 80-plus individuals. The norms and cut-off scores for mild cognitive impairment (MCI) and dementia of the MoCA are different among five Chinese versions. OBJECTIVE: To provide the cut-off scores in detecting MCI and dementia of the Peking Medical Union College Hospital version of the MoCA (MoCA-P). METHODS: In a cross-sectional survey, Chinese veterans aged ≥60 years completed the MoCA-P and the Mini-Mental State Examination (MMSE). RESULTS: Among 7,445 elderly veterans, 5,085 (68.30%) were aged ≥80 years old, 2,621 (35.20%) had 6 years of education or less, 6,847 (91.97%) were male, and 2,311 (31.04%) and 984 (13.22%) veterans were diagnosed as having MCI and dementia, respectively. Adding two points and one point to the MoCA scores for the primary and middle school groups, respectively, can fully adjust for the notable impact of education but cannot compensate for the effect of age. In the three age groups (60-79, 80-89, and ≥90 years old), the optimal MoCA-P cut-off scores for detecting MCI were ≤25, ≤24, and ≤23, respectively, and for detecting dementia were ≤24, ≤21, and ≤19, respectively, which demonstrated relatively high sensitivities and specificities. The areas under the curves for the MoCA-P for detecting MCI and dementia (0.937 and 0.908, respectively) were greater than those for the MMSE (0.848 and 0.892, respectively). CONCLUSION: Compared with the MMSE, the MoCA-P is significantly better for detecting MCI in the elderly, particularly in the oldest old population, and it also displays more effectiveness in detecting dementia.

Relation of ABO blood groups to the severity of coronary atherosclerosis: an Gensini score assessment.

OBJECTIVE: Although the study on the relationship between ABO blood groups and coronary atherosclerosis has a long history, few data is available regarding ABO to severity of coronary atherosclerosis in a large cohort study. Therefore, the present study aimed to investigate the relation of the ABO blood groups to the severity of coronary atherosclerosis assessed by Gensini score (GS) in a large Chinese cohort undergoing coronary angiography. METHODS: A total of 2919 consecutive patients undergoing coronary angiography were enrolled, and their baseline characteristics and ABO blood groups were collected. The GS was calculated as 1st tertile (0-10), 2nd tertile (11-36), 3rd tertile (>36) according to angiographic results. The relation of the ABO blood groups to GS was investigated. RESULTS: The frequency of blood group A was significantly higher in the upper GS tertiles (24.4% vs. 28.2% vs. 29.5%, p = 0.032). Multivariable linear regression analysis revealed that blood group A was independently associated with GS (ß = 0.043, p = 0.017). Likewise, multivariable logistic regression analysis showed that group A remained significantly associated with mid-high GS (OR = 1.44, 95% CI 1.16-1.80, p = 0.001), and the group O was showed as a protective factor (OR = 0.77, 95% CI = 0.65-0.92, p = 0.004). CONCLUSION: In this large Chinese cohort study, the data indicated that there was an association between ABO blood groups and the severity of coronary atherosclerosis. Moreover, the blood group A was an independent risk factor for serious coronary atherosclerosis.

Usefulness of the neutrophil-to-lymphocyte ratio in predicting the severity of coronary artery disease: a Gensini score assessment.

AIM: The usefulness of the white blood cell (WBC) count and neutrophil-to-lymphocyte ratio (NLR) in predicting the severity of stable coronary artery disease (CAD) has not been sufficiently evaluated, particularly based on strict coronary assessments. The aim of the present study was to investigate the WBC count and NLR in predicting the severity of angiographically proven CAD. METHODS: A total of 2,976 CAD patients and 571 non-CAD patients were consecutively enrolled, and the CAD patients were classified into the three groups according to the tertile of the Gensini score (GS, low GS<18, n=989; intermediate GS 18-41, n=995 and high GS>41, n=992). The efficacy of the WBC count and NLR in predicting the risk and severity of CAD as well as the correlations between these markers and the GS were analyzed. A receiver operating characteristic (ROC) curve analysis was also performed. RESULTS: The NLR was found to be an independent predictor of both the presence of CAD (OR=1.18, 95%CI: 1.09-1.27, p=0.009) and a high GS (OR=1.10, 95%CI: 1.01-1.16, p=0.032). In addition, there were mild positive correlations between the GS and the NLR, WBC and proportions of neutrophils and monocytes. In the ROC curves analysis, the NLR was found to have the largest area under the curve (AUC=0.63, 95%CI: 0.59-0.67, p=0.000), with an optimal cut-off value of 2.04 (sensitivity: 62.1%, specificity: 54.8%) for predicting a high GS. CONCLUSIONS: The NLR is a valuable independent predictor of the severity of CAD assessed according to the GS. In particular, an NLR of >2.04 indicates a higher risk of CAD and greater severity of CAD lesions.