ObjectiveAngelicae Sinensis Radix， a commonly used medicinal herb with both medicinal and edible properties， is frequently adulterated in the market， severely affecting the clinical efficacy of preparations. While qualitative identification techniques for adulterants and counterfeits are now relatively mature， quantitative detection methods for adulterated processed products remain unexplored. Quantitative detection research of Angelicae Sinensis Radix and its primary closely related adulterant， "Tu Danggui" （Angelica gigas）， was conducted to establish a herbal quantitative molecular detection （Herb-Q） method for Angelicae Sinensis Radix and its processed products， providing a model for the establishment of quantitative detection technologies for Angelicae Sinensis Radix and related health products. MethodsThe specific single-nucleotide polymorphism （SNP） loci of Angelicae Sinensis Radix and Angelica gigas Nakai were screened based on the complete chloroplast genome sequence. The specific SNP loci of Angelicae Sinensis Radix were selected for quantitative methodological investigations （linearity， limit of quantification， limit of detection， and reproducibility） by mixing the powder of the herbs with different adulteration ratios. Huoxue Zhitong powder with three distinct adulteration ratios （15%， 25%， and 35%） was utilized to ascertain the precision of the Herb-Q method for the quantitative detection of Chinese patent medicines containing Angelicae Sinensis Radix. ResultsBy comparing the 123 chloroplast genome sequences of Angelicae Sinensis Radix， based on the principles of intraspecies conservation， interspecies specificity， and meeting the requirements of pyrophosphate high-throughput sequencing， it was determined that 9 674^th locus （A/G） in the chloroplast genome sequence NC_042826.1 and 38 592^nd locus （T/C） in the chloroplast genome sequence NC_029393.1 could be the exclusive molecular identification loci of Angelicae Sinensis Radix and Angelica gigas Nakai， respectively. The linear relationship R² of the Herb-Q method established by selecting the specific 9 674^th locus （A/G） of Angelicae Sinensis Radix was 0.997 4 （R²>0.99）， indicating an excellent linear relationship. The limits of quantification and detection were established at 2.0%， exhibiting excellent reproducibility ［relative standard deviation（RSD）<2.0%］. The established quantitative system based on the Herb-Q method detected the adulteration amount of counterfeit A. gigas in the Huoxue Zhitong powder， with an average deviation of 1.3% for three molecular quantitative replicates. ConclusionThis research demonstrates that the Herb-Q quantitative detection method established based on the 9 674^th locus （A/G） in the chloroplast genome sequence NC_042826.1 of Angelicae Sinensis Radix has good applicability， objectivity， and accuracy for Angelicae Sinensis Radix and A. gigas， and its processed products. This method has the capacity to provide technical support for the quantitative detection of commercially available Angelicae Sinensis Radix derivatives， including traditional Chinese medicinal preparations， dietary supplements， and nutraceuticals.

ObjectiveAngelicae Sinensis Radix， a commonly used medicinal herb with both medicinal and edible properties， is frequently adulterated in the market， severely affecting the clinical efficacy of preparations. While qualitative identification techniques for adulterants and counterfeits are now relatively mature， quantitative detection methods for adulterated processed products remain unexplored. Quantitative detection research of Angelicae Sinensis Radix and its primary closely related adulterant， "Tu Danggui" （Angelica gigas）， was conducted to establish a herbal quantitative molecular detection （Herb-Q） method for Angelicae Sinensis Radix and its processed products， providing a model for the establishment of quantitative detection technologies for Angelicae Sinensis Radix and related health products. MethodsThe specific single-nucleotide polymorphism （SNP） loci of Angelicae Sinensis Radix and Angelica gigas Nakai were screened based on the complete chloroplast genome sequence. The specific SNP loci of Angelicae Sinensis Radix were selected for quantitative methodological investigations （linearity， limit of quantification， limit of detection， and reproducibility） by mixing the powder of the herbs with different adulteration ratios. Huoxue Zhitong powder with three distinct adulteration ratios （15%， 25%， and 35%） was utilized to ascertain the precision of the Herb-Q method for the quantitative detection of Chinese patent medicines containing Angelicae Sinensis Radix. ResultsBy comparing the 123 chloroplast genome sequences of Angelicae Sinensis Radix， based on the principles of intraspecies conservation， interspecies specificity， and meeting the requirements of pyrophosphate high-throughput sequencing， it was determined that 9 674^th locus （A/G） in the chloroplast genome sequence NC_042826.1 and 38 592^nd locus （T/C） in the chloroplast genome sequence NC_029393.1 could be the exclusive molecular identification loci of Angelicae Sinensis Radix and Angelica gigas Nakai， respectively. The linear relationship R² of the Herb-Q method established by selecting the specific 9 674^th locus （A/G） of Angelicae Sinensis Radix was 0.997 4 （R²>0.99）， indicating an excellent linear relationship. The limits of quantification and detection were established at 2.0%， exhibiting excellent reproducibility ［relative standard deviation（RSD）<2.0%］. The established quantitative system based on the Herb-Q method detected the adulteration amount of counterfeit A. gigas in the Huoxue Zhitong powder， with an average deviation of 1.3% for three molecular quantitative replicates. ConclusionThis research demonstrates that the Herb-Q quantitative detection method established based on the 9 674^th locus （A/G） in the chloroplast genome sequence NC_042826.1 of Angelicae Sinensis Radix has good applicability， objectivity， and accuracy for Angelicae Sinensis Radix and A. gigas， and its processed products. This method has the capacity to provide technical support for the quantitative detection of commercially available Angelicae Sinensis Radix derivatives， including traditional Chinese medicinal preparations， dietary supplements， and nutraceuticals.

Cardiac fibrosis is characterized by an elevated amount of extracellular matrix (ECM) within the heart. However, the persistence of cardiac fibrosis ultimately diminishes contractility and precipitates cardiac dysfunction. Circular RNAs (circRNAs) are emerging as important regulators of cardiac fibrosis. Here, we elucidate the functional role of a specific circular RNA CELF1 in cardiac fibrosis and delineate a novel feedback loop mechanism. Functionally, circ-CELF1 was involved in enhancing fibrosis-related markers' expression and promoting the proliferation of cardiac fibroblasts (CFs), thereby exacerbating cardiac fibrosis. Mechanistically, circ-CELF1 reduced the ubiquitination-degradation rate of BRPF3, leading to an elevation of BRPF3 protein levels. Additionally, BRPF3 acted as a modular scaffold for the recruitment of histone acetyltransferase KAT7 to facilitate the induction of H3K14 acetylation within the promoters of the Celf1 gene. Thus, the transcription of Celf1 was dramatically activated, thereby inhibiting the subsequent response of their downstream target gene Smad7 expression to promote cardiac fibrosis. Moreover, Celf1 further promoted Celf1 pre-mRNA transcription and back-splicing, thereby establishing a feedback loop for circ-CELF1 production. Consequently, a novel feedback loop involving CELF1/circ-CELF1/BRPF3/KAT7 was established, suggesting that circ-CELF1 may serve as a potential novel therapeutic target for cardiac fibrosis.

When bacteria inhabit an unfavorable environment, they adapt by forming small colony variants (SCVs) under selective pressure, resulting in a slow-growing subpopulation with distinct phenotypic and pathogenic characteristics. Phenotypically, SCVs exhibit a slow growth rate, atypical colony morphology, and unusual biochemical traits. Clinically, SCVs demonstrate reduced susceptibility to antibiotics and can persistently proliferate within the host environment as potential pathogens, posing a significant challenge for the treatment of associated infections. This paper analyzes the phenotypic, genetic, and clinical characteristics of SCVs to provide a reference for the clinical diagnosis and treatment of SCV infections.

When bacteria inhabit an unfavorable environment, they adapt by forming small colony variants (SCVs) under selective pressure, resulting in a slow-growing subpopulation with distinct phenotypic and pathogenic characteristics. Phenotypically, SCVs exhibit a slow growth rate, atypical colony morphology, and unusual biochemical traits. Clinically, SCVs demonstrate reduced susceptibility to antibiotics and can persistently proliferate within the host environment as potential pathogens, posing a significant challenge for the treatment of associated infections. This paper analyzes the phenotypic, genetic, and clinical characteristics of SCVs to provide a reference for the clinical diagnosis and treatment of SCV infections.

Objective:To analyze the epidemiological distribution characteristics, influencing factors, and infection rates of pertussis in the population of Henan Province.Methods:From 2022 to 2023, a cross-sectional survey was conducted to investigate the permanent population in Henan Province. Enzyme-linked immunosorbent assay (ELISA) was used to detect anti-pertussis toxin IgG (PT-IgG), analyze the antibody positivity rate (≥20 IU/ml) and median concentration (MC), and estimate the pertussis infection rate based on PT IgG ≥40 IU/ml. The rank sum test was used to compare antibody levels among groups, and the χ2 test was used to compare antibody positive rates and infection rates among groups. Results:A total of 4 810 research subjects were included in this study. The overall positive rate of PT-IgG was 12.10% and MC was 3.04 (0.35, 10.36) IU/ml. There were significant differences both in positive rates and antibody levels of PT-IgG among different regions or age groups (region positive rate: χ2=134.06, P<0.001, MC: H=337.74, P<0.001; age group positive rate: χ2=45.27, P<0.001, MC: H=134.49, P<0.001). Both the positive rate of PT-IgG (25.26%) and MC (8.01 IU/ml) were the highest within one year after completing a full course of vaccination. There were significant differences in positive rates and antibody levels among people receiving different types of pertussis vaccines (positive rate: χ2=12.38, P=0.006, MC: H=17.93, P<0.001). The antibody positivity rate (35.71%) and MC (8.88 IU/ml) of the people who received cell-free pertussis inactivated poliomyelitis influenza type b (combined) vaccine throughout the course were higher than those who received other types of vaccines. The natural infection rate of pertussis was evaluated for individuals aged≥3 years who had no history of pertussis vaccine immunization within the year prior to sampling. With a high vaccination rate, the estimated infection rate of pertussis in the population was 5 757.22/100 000. The infection rates in the 3-year-old (1 940.16/100 000) and 4-year-old (1 765.68/100 000) populations were at a low level among the entire population, reaching their peak at the age of 6 (12 656.71/100 000). Subsequently, although the infection rate continued to decline, it remained at a high level and peaked again at the age of 40-49 years (8 740.39/100 000). There was a statistically significant difference in the estimated infection rate of pertussis among different age groups ( χ2=53.21, P<0.001). Conclusion:The PT-IgG level of pertussis in the population of Henan Province is generally at a low level. The estimated infection rate of pertussis is much higher than the reported incidence rate. A booster dose of pertussis vaccine is recommended at 6 years old.

Objective:To analyze the epidemiological distribution characteristics, influencing factors, and infection rates of pertussis in the population of Henan Province.Methods:From 2022 to 2023, a cross-sectional survey was conducted to investigate the permanent population in Henan Province. Enzyme-linked immunosorbent assay (ELISA) was used to detect anti-pertussis toxin IgG (PT-IgG), analyze the antibody positivity rate (≥20 IU/ml) and median concentration (MC), and estimate the pertussis infection rate based on PT IgG ≥40 IU/ml. The rank sum test was used to compare antibody levels among groups, and the χ2 test was used to compare antibody positive rates and infection rates among groups. Results:A total of 4 810 research subjects were included in this study. The overall positive rate of PT-IgG was 12.10% and MC was 3.04 (0.35, 10.36) IU/ml. There were significant differences both in positive rates and antibody levels of PT-IgG among different regions or age groups (region positive rate: χ2=134.06, P<0.001, MC: H=337.74, P<0.001; age group positive rate: χ2=45.27, P<0.001, MC: H=134.49, P<0.001). Both the positive rate of PT-IgG (25.26%) and MC (8.01 IU/ml) were the highest within one year after completing a full course of vaccination. There were significant differences in positive rates and antibody levels among people receiving different types of pertussis vaccines (positive rate: χ2=12.38, P=0.006, MC: H=17.93, P<0.001). The antibody positivity rate (35.71%) and MC (8.88 IU/ml) of the people who received cell-free pertussis inactivated poliomyelitis influenza type b (combined) vaccine throughout the course were higher than those who received other types of vaccines. The natural infection rate of pertussis was evaluated for individuals aged≥3 years who had no history of pertussis vaccine immunization within the year prior to sampling. With a high vaccination rate, the estimated infection rate of pertussis in the population was 5 757.22/100 000. The infection rates in the 3-year-old (1 940.16/100 000) and 4-year-old (1 765.68/100 000) populations were at a low level among the entire population, reaching their peak at the age of 6 (12 656.71/100 000). Subsequently, although the infection rate continued to decline, it remained at a high level and peaked again at the age of 40-49 years (8 740.39/100 000). There was a statistically significant difference in the estimated infection rate of pertussis among different age groups ( χ2=53.21, P<0.001). Conclusion:The PT-IgG level of pertussis in the population of Henan Province is generally at a low level. The estimated infection rate of pertussis is much higher than the reported incidence rate. A booster dose of pertussis vaccine is recommended at 6 years old.

Objective:To analyze the histopathological features of primary aldosteronism(PA) based on the histopathology of primary aldosteronism(HISTALDO) consensus classification in patients undergoing unilateral adrenalectomy and explore its correlation with clinical characteristics and postoperative outcomes.Methods:A retrospective study was conducted on 105 unilateral PA patients treated at the First Affiliated Hospital of Nanjing Medical University between April 2017 and April 2022. Postoperative histopathology was classified according to the latest HISTALDO consensus and compared with traditional classifications. Differences in clinical features and primary aldosteronism surgical outcomes(PASO) were analyzed across classifications.Results:HISTALDO classification showed that 67.6% of patients were classical subtype and 32.4% were non-classical subtype. Compared to the non-classical group, the classical group showed lower age, body mass index, serum potassium concentration, fasting blood glucose, HbA 1C, and proportion of CYP11B2 negative nodules as well as higher systolic blood pressure, diastolic blood pressure, prevalence of hypokalemia, and 24-hour urine potassium(all P<0.05). Based on PASO criteria, complete clinical and biochemical success were achieved in 41.7% and 83.3% subjects respectively. A significantly higher proportion of classical group displayed complete clinical and biochemical success than nonclassical group(all P<0.05). Multivariate logistic regression analysis showed that lower serum potassium concentration( OR=0.15, 95% CI 0.03-0.84, P<0.05) and proportion of CYP11B2 negative nodules( OR=0.20, 95% CI 0.05-0.86, P<0.05) were related to classical pathological type, and classical pathological type was independently related to clinical and biochemical remission(all P<0.05). Conclusion:Standardized HISTALDO histopathological classification enhances diagnostic accuracy and predicts postoperative outcomes in unilateral PA, providing a foundation for precise stratified management of PA patients.

Objective:To analyze the histopathological features of primary aldosteronism(PA) based on the histopathology of primary aldosteronism(HISTALDO) consensus classification in patients undergoing unilateral adrenalectomy and explore its correlation with clinical characteristics and postoperative outcomes.Methods:A retrospective study was conducted on 105 unilateral PA patients treated at the First Affiliated Hospital of Nanjing Medical University between April 2017 and April 2022. Postoperative histopathology was classified according to the latest HISTALDO consensus and compared with traditional classifications. Differences in clinical features and primary aldosteronism surgical outcomes(PASO) were analyzed across classifications.Results:HISTALDO classification showed that 67.6% of patients were classical subtype and 32.4% were non-classical subtype. Compared to the non-classical group, the classical group showed lower age, body mass index, serum potassium concentration, fasting blood glucose, HbA 1C, and proportion of CYP11B2 negative nodules as well as higher systolic blood pressure, diastolic blood pressure, prevalence of hypokalemia, and 24-hour urine potassium(all P<0.05). Based on PASO criteria, complete clinical and biochemical success were achieved in 41.7% and 83.3% subjects respectively. A significantly higher proportion of classical group displayed complete clinical and biochemical success than nonclassical group(all P<0.05). Multivariate logistic regression analysis showed that lower serum potassium concentration( OR=0.15, 95% CI 0.03-0.84, P<0.05) and proportion of CYP11B2 negative nodules( OR=0.20, 95% CI 0.05-0.86, P<0.05) were related to classical pathological type, and classical pathological type was independently related to clinical and biochemical remission(all P<0.05). Conclusion:Standardized HISTALDO histopathological classification enhances diagnostic accuracy and predicts postoperative outcomes in unilateral PA, providing a foundation for precise stratified management of PA patients.

OBJECTIVE: To investigate the variations in the expression of voltage-gated sodium (Na_v) channel subunits during development of rat cerebellar Purkinje neurons and their correlation with maturation of electrophysiological characteristics of the neurons. METHODS: We observed the changes in the expression levels of Na_V1.1, 1.2, 1.3 and 1.6 during the development of Purkinje neurons using immunohistochemistry in neonatal (5-7 days after birth), juvenile (12-14 days), adolescent (21-24 days), and adult (42-60 days) SD rats. Using whole-cell patch-clamp technique, we recorded the spontaneous electrical activity of the neurons in ex vivo brain slices of rats of different ages to analyze the changes of electrophysiological characteristics of these neurons during development. RESULTS: The expression of Na_V subunits in rat cerebellar Purkinje neurons showed significant variations during development. Na_V1.1 subunit was highly expressed throughout the developmental stages and increased progressively with age (P < 0.05). Na_V1.2 expression was not detected in the neurons in any of the developmental stages (P > 0.05). The expression level of Na_V1.3 decreased with development and became undetectable after adolescence (P < 0.05). Na_V1.6 expression was not detected during infancy, but increased with further development (P < 0.05). Na_V1.1 and Na_V1.3 were mainly expressed in the early stages of development. With the maturation of the rats, Na_V1.3 expression disappeared and Na_V1.6 expression increased in the neurons. Na_V1.1 and Na_V1.6 were mainly expressed after adolescence. The total Na_V protein level increased gradually with development (P < 0.05) and tended to stabilize after adolescence. The spontaneous frequency and excitability of the Purkinje neurons increased gradually with development and reached the mature levels in adolescence. The developmental expression of Na_V subunits was positively correlated with discharge frequency (r=0.9942, P < 0.05) and negatively correlated with the excitatory threshold of the neurons (r=0.9891, P < 0.05). CONCLUSION The changes in the expression levels of Na_V subunits are correlated with the maturation of high frequency electrophysiological properties of the neurons, suggesting thatmature Na_V subunit expressions is the basis of maturation of electrophysiological characteristics of the neurons.
Rats ; Animals ; Purkinje Cells/physiology* ; Rats, Sprague-Dawley ; Neurons ; Brain ; Sodium/metabolism*