BACKGROUND:Carnosic acid,a bioactive compound found in rosemary,has been shown to reduce inflammation and reactive oxygen species(ROS).However,its mechanism of action in osteoclast differentiation remains unclear. OBJECTIVE:To investigate the effects of carnosic acid on osteoclast activation,ROS production,and mitochondrial function. METHODS:Primary bone marrow-derived macrophages from mice were extracted and cultured in vitro.Different concentrations of carnosic acid(0,10,15,20,25 and 30 μmol/L)were tested for their effects on bone marrow-derived macrophage proliferation and toxicity using the cell counting kit-8 cell viability assay to determine a safe concentration.Bone marrow-derived macrophages were cultured in graded concentrations and induced by receptor activator of nuclear factor-κB ligand for osteoclast differentiation for 5-7 days.The effects of carnosic acid on osteoclast differentiation and function were then observed through tartrate-resistant acid phosphatase staining,F-actin staining,H2DCFDA probe and mitochondrial ROS,and Mito-Tracker fluorescence detection.Western blot and RT-PCR assays were subsequently conducted to examine the effects of carnosic acid on the upstream and downstream proteins of the receptor activator of nuclear factor-κB ligand-induced MAPK signaling pathway. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining and F-actin staining showed that carnosic acid dose-dependently inhibited in vitro osteoclast differentiation and actin ring formation in the cell cytoskeleton,with the highest inhibitory effect observed in the high concentration group(30 μmol/L).Carnosic acid exhibited the most significant inhibitory effect during the early stages(days 1-3)of osteoclast differentiation compared to other intervention periods.Fluorescence imaging using the H2DCFDA probe,mitochondrial ROS,and Mito-Tracker demonstrated that carnosic acid inhibited cellular and mitochondrial ROS production while reducing mitochondrial membrane potential,thereby influencing mitochondrial function.The results of western blot and RT-PCR revealed that carnosic acid could suppress the expression of NFATc1,CTSK,MMP9,and C-fos proteins associated with osteoclast differentiation,and downregulate the expression of NFATc1,Atp6vod2,ACP5,CTSK,and C-fos genes related to osteoclast differentiation.Furthermore,carnosic acid enhanced the expression of antioxidant enzyme proteins and reduced the generation of ROS during the process of osteoclast differentiation.Overall,carnosic acid exerts its inhibitory effects on osteoclast differentiation by inhibiting the phosphorylation modification of the P38/ERK/JNK protein and activating the MAPK signaling pathway in bone marrow-derived macrophages.

Jiegengtang is a basic formula for treating sore throat and cough. By means of bibliometrics， this study conducted a textual research and analysis on the key information such as formula origin， decocting methods， and clinical application of Jiegengtang. After the research， it can be seen that Jiegengtang is firstly contained in Treatise on Febrile and Miscellaneous Disease， which is also known as Ganjietang， and it has been inherited and innovated by medical practitioners of various dynasties in later times. The origins of Chinese medicines in this formula is basically clear， Jiegeng is the dried roots of Platycodon grandiflorum， Gancao is the dried roots and rhizomes of Glycyrrhiza uralensis， the two medicines are selected raw products. The dosage is 27.60 g of Glycyrrhizae Radix et Rhizoma and 13.80 g of Platycodonis Radix， decocted with 600 mL of water to 200 mL， taken warmly after meals， twice a day， 100 mL for each time. In ancient times， Jiegengtang was mainly used for treating Shaoyin-heat invasion syndrome， with cough and sore throat as its core symptoms. In modern clinical practice， Jiegengtang is mainly used for respiratory diseases such as pharyngitis， esophagitis， tonsillitis and lung abscess， especially for pharyngitis and lung abscess with remarkable efficacy. This paper can provide literature reference basis for the modern clinical application and new drug development of Jiegengtang.

Jiegengtang is a basic formula for treating sore throat and cough. By means of bibliometrics， this study conducted a textual research and analysis on the key information such as formula origin， decocting methods， and clinical application of Jiegengtang. After the research， it can be seen that Jiegengtang is firstly contained in Treatise on Febrile and Miscellaneous Disease， which is also known as Ganjietang， and it has been inherited and innovated by medical practitioners of various dynasties in later times. The origins of Chinese medicines in this formula is basically clear， Jiegeng is the dried roots of Platycodon grandiflorum， Gancao is the dried roots and rhizomes of Glycyrrhiza uralensis， the two medicines are selected raw products. The dosage is 27.60 g of Glycyrrhizae Radix et Rhizoma and 13.80 g of Platycodonis Radix， decocted with 600 mL of water to 200 mL， taken warmly after meals， twice a day， 100 mL for each time. In ancient times， Jiegengtang was mainly used for treating Shaoyin-heat invasion syndrome， with cough and sore throat as its core symptoms. In modern clinical practice， Jiegengtang is mainly used for respiratory diseases such as pharyngitis， esophagitis， tonsillitis and lung abscess， especially for pharyngitis and lung abscess with remarkable efficacy. This paper can provide literature reference basis for the modern clinical application and new drug development of Jiegengtang.

Objective To explore the polymorphism of tyrosinase (TYR) and melanocortin 1 receptor (MC1R) genes and their mRNA expression levels in relation to coat-color phenotypes in guinea pigs, providing genetic markers for locating dominant traits in guinea pigs. Methods A total of 57 self-bred ordinary-level guinea pigs were selected and divided into three groups based on coat color: white (n=22), variegated (n=22) and black (n=13). The guinea pigs were euthanized with an overdose of pentobarbital sodium via intraperitoneal injection. DNA was then extracted from the dorsal skin tissue. Polymorphism in the coding sequence (CDS) of the exons of the TYR and MC1R genes in each group was detected by cloning and sequencing. The mRNA expression of the two genes in skin tissues was detected by real-time fluorescent quantitative PCR to investigate the relationship between these genes and guinea pig coat color. Results A single nucleotide polymorphism (SNP) site was found in the CDS region of TYR exon Ⅰ, where the base A was replaced by G. All white guinea pigs had the G/G genotype for TYR, while no deep-colored (variegated and black) guinea pigs exhibited the G/G genotype for TYR. Most deep-colored guinea pigs had the A/A genotype, and a few had A/G genotype. The A/A genotype frequency in black guinea pigs was higher than in variegated guinea pigs. A 2 760 bp sequence deletion was identified in the exon of the MC1R gene, marked as the - gene, with non-deleted samples marked as N gene. Most white guinea pigs had the -/- genotype for MC1R, variegated guinea pigs mainly had the -/N genotype, and black guinea pigs mainly had the N/N genotype, with a few showing the -/N. The TYR gene expression level was higher in white guinea pigs, lower in variegated guinea pigs, and intermediate in black guinea pigs, but there was no significant difference among the three groups (P>0.05). The MC1R gene expression level in white guinea pigs was extremely low, while both variegated and black guinea pigs showed significantly higher levels than white guinea pigs (P<0.01). Black guinea pigs showed significantly higher levels than variegated guinea pigs (P<0.05). ConclusionThe TYR and MC1R genes synergistically regulate coat color of guinea pigs. The G-site mutation in the TYR gene may lead to albinism, and the change of N-site in the MC1R gene affects the depth of the coat color.

The scientific evaluation of the construction and development of university disciplines is an important part and a key challenge in China’s “Double First-Class” initiative. Based on the ESI and InCites databases, this study conducted a year-long observation and analysis of the disciplines at universities in Jiangsu Province under the “Double First-Class” framework. The research focused on the development trends of potential and emerging disciplines, while exploring the influence of traditional predominant disciplines on the growth of these emerging fields and the pathways through which such influence might occur. The study aims to provide some reference and insights for the development of disciplines in the universities in Jiangsu. The findings reveal that, overall, the discipline development maturity of the universities is high, with a positive growth trend and significant development potential in certain fields, which are strongly supported by the traditional predominant disciplines. However, an imbalance in the development across various discipline categories has also been observed. Looking ahead, universities are encouraged to capitalize on their unique academic strengths to foster collaborative development, emphasize the growth of social science disciplines, promote interdisciplinary integration, and cultivate high-level talent for society. This approach will better facilitate the establishment of high-level universities and contribute to the realization of the goal of building a strong educational nation.

ObjectiveFunctional near-infrared spectroscopy (fNIRS), a novel non-invasive technique for monitoring cerebral activity, can be integrated with upper limb rehabilitation robots to facilitate the real-time assessment of neurological rehabilitation outcomes. The rehabilitation robot is designed with 3 training modes: passive, active, and resistance. Among these, the resistance mode has been demonstrated to yield superior rehabilitative outcomes for patients with a certain level of muscle strength. The control modes in the resistance mode can be categorized into dynamic and static control. However, the effects of different control modes in the resistance mode on the motor function of patients with upper limb hemiplegia in stroke remain unclear. Furthermore, the effects of force, an important parameter of different control modes, on the activation of brain regions have rarely been reported. This study investigates the effects of dynamic and static resistance modes under varying resistance levels on cerebral functional alterations during motor rehabilitation in post-stroke patients. MethodsA cohort of 20 stroke patients with upper limb dysfunction was enrolled in the study, completing preparatory adaptive training followed by 3 intensity-level tasks across 2 motor paradigms. The bilateral prefrontal cortices (PFC), bilateral primary motor cortices (M1), bilateral primary somatosensory cortices (S1), and bilateral premotor and supplementary motor cortices (PM) were examined in both the resting and motor training states. The lateralization index (LI), phase locking value (PLV), network metrics were employed to examine cortical activation patterns and topological properties of brain connectivity. ResultsThe data indicated that both dynamic and static modes resulted in significantly greater activation of the contralateral M1 area and the ipsilateral PM area when compared to the resting state. The static patterns demonstrated a more pronounced activation in the contralateral M1 in comparison to the dynamic patterns. The results of brain network analysis revealed significant differences between the dynamic and resting states in the contralateral PFC area and contralateral M1 area (F=4.709, P=0.038), as well as in the contralateral PM area and ipsilateral M1 area (F=4.218, P=0.049). Moreover, the findings indicated a positive correlation between the activation of the M1 region and the increase in force in the dynamic mode, which was reversed in the static mode. ConclusionBoth dynamic and static resistance training modes have been demonstrated to activate the corresponding brain functional regions. Dynamic resistance modes elicit greater oxygen changes and connectivity to the region of interest (ROI) than static resistance modes. Furthermore, the effects of increasing force differ between the two modes. In patients who have suffered a stroke, dynamic modes may have a more pronounced effect on the activation of exercise-related functional brain regions.

Objective:To investigate the predictive role of relative diffusion-weighted imaging (DWI) signal intensity (DWI-rSI) in outcome in patients with anterior circulation large vessel occlusion cardioembolic stroke and successful recanalization after endovascular therapy (EVT).Methods:Patients with anterior circulation large vessel occlusion stroke due to cardioembolic embolism underwent EVT and successful recanalization at the Affiliated Hospital of Yangzhou University from March 2017 to March 2023 were retrospectively included. According to the modified Rankin Scale score 3 months after procedure, the patients were divided into a good outcome group (0-2 points) and a poor outcome group (3-6 points). Multivariate logistic regression analysis was used to identify independent predictive factors for poor outcome. Results:A total of 59 patients were enrolled, including 29 males (49.2%), median age of 74 years (interquartile range, 68-80 years). The median baseline National Institutes of Health Stroke Scale (NIHSS) score was 15 (12-21), and the median DWI Alberta Stroke Program Early CT Score (ASPECTS) was 8 (5-9). Thirty-two patients (54.2%) had good outcome, and 27 (45.8%) had poor outcome. Among them, 9 patients (15.3%) died (6 died from cerebral herniation after malignant brain edema, 2 died from complications, and 1 died from severe intracranial hemorrhage after procedure). Twenty-one patients (35.6%) experienced hemorrhagic transformation, including 12 (20.3%) with symptomatic intracranial hemorrhage. There were significant differences in baseline systolic blood pressure, NIHSS score, DWI-ASPECTS, DWI-rSI, and incidence of symptomatic intracranial hemorrhage between the good outcome group and the poor outcome group (all P<0.05). Multivariate logistic regression analysis showed that baseline systolic blood pressure (odds ratio 0.977, 95% confidence interval 0.919-0.991; P=0.015) and DWI-rSI (odds ratio 11.809, 95% confidence interval 1.932-72.170; P=0.008) were the independent predictors for poor outcome. Conclusion:DWI-rSI can predict the outcome of patients with anterior circulation large vessel occlusion cardioembolic stroke and successful recanalization after EVT.

Aim To explore the antimicrobial activity of a novel R-type phage tail-like bacteriocin(PTLB)secreted by Enterobacter cloacae SHAMU191747 a-gainst methicillin-resistant Staphylococcus aureus(MR-SA).Methods Antagonistic activity of E.cloacae SHAMU191747 against MRS A was detected by LB agar plate antagonistic test and LB broth micro-fermentation test.The crude extract of fermentation supernatant of E.cloacae SHAMU191747 was prepared by ultra-high-speed centrifugation and density gradient centrifuga-tion.The novel R-type PTLB in the crude extract was detected by transmission electron microscopy.The an-timicrobial activity of the crude extract against MRSA was verified by LB agar plate spot-seeding method.The molecular weight of the novel R-type PTLB was detected by SDS-PAGE electrophoresis.Results E.cloacae SHAMU191747 secreted a novel R-type PTLB,and had a strong antagonistic effect on MRSA.The no-vel R-type PTLB had a molecular weight of approxi-mately 35 ku and could efficiently kill MRSA.The physical dimensions of its tail-sheath-uncontracted functional molecules were(142.7±4.3)×(13.8±0.6)nm,and those of the tail-sheath-contracted non-functional molecules were(57.7±1.2)×(20.8±1.5)nm.Conclusions The novel R-type PTLB pro-duced by E.cloacae SHAMU191747 can efficiently kill MRSA,and has the potential to be developed into a novel antimicrobial drug with great prospects for clini-cal application.

Objective:To study the quality and stability of commercial ready-to-use tryptone soya agar(TSA)after storing at 2-25 ℃ for different storage duration under dark condition in order to discuss a determination method of validity period for medium.Methods:Three consecutive batches of ready-to-use TSA medium from two manufac-turers were selected and stored at 2-25 ℃ under dark conditions for 30,90 and 180 days,respectively.The appearance,pH,medium suitability and sterility of the medium were tested.Results:The results of appearance,pH,suitability and sterility of TSA medium from two manufacturers for each batch under different storage duration all met the requirements of the Chinese Pharmacopoeia 2020 Volume IV on the quality control of medium.Conclusion:The TSA medium from two manufacturers all met the requirements when stored for 180 days at 2-25 ℃ under dark condition,indicating that the validity period of TSA medium from two manufacturers can reach 180 days.

Objective To analyze the diagnostic value of ultrasound blood quantitative parameters combined with murine sarcoma filtering toxin oncogenic homolog B(BRAF)gene mutation in Hashimoto's thyroiditis(HT)with thyroid carcinoma.Methods A total of 86 patients with HT and thyroid nodules in Sir Run Run Hospital,Nanjing Medical University from June 2020 to June 2022 were selected as the research objects.The patients with thyroid cancer were included into the malignant group(n=26),and the patients with benign thyroid nodules were included into the benign group(n=60)according to the pathological examination results.The patients in the malignant group was divided into the metastasis group(n=10)and the non-metastasis group(n=16)according to whether lymph node metastasis occurred.All subjects underwent color Doppler ultrasonography and BRAF gene mutation detection.The pulsatility index(PI),resistance index(RI),peak systolic velocity(S),end diastolic velocity(D),peak systolic velocity to end diastolic velocity ratio(S/D)and BRAF gene mutation between the malignant group and the benign group were compared.The detection rate of blood flow signals by CDFI,the detection rate of blood flow signals of grade 2 to 3 and the mutation of BRAF gene between the metastasis group and the non-metastasis group were compared.The value of the above blood parameters and BRAF gene mutations in the diagnosis of HT complicated with thyroid cancer was evaluated by receiver operating characteristic(ROC)curve,and the value of the above blood parameters and BRAF gene mutation in the diagnosis of lymph node metastasis of thyroid cancer was evaluated.Results The PI,RI,S,D,S/D and positive rate of BRAF gene mutation in the malignant group were significantly higher than those in the benign group(P<0.05).The detection rate of blood flow signals by CDFI,the detection rate of blood flow signals of grade 2 to 3,and the positive rate of BRAF gene mutation in the metastasis group were significantly higher than those in the non-metastasis group(P<0.05).ROC curve analysis showed that the area under the curve(AUC)of RI,PI,S,D,S/D and BRAF gene mutation in the combined assessment of HT complicated with thyroid cancer was 0.951,with the sensitivity of 88.46%and the specificity of 93.33%,and the efficiency of combined assessment was better than that of the independent assessment of each index.Meanwhile,the AUC of the detection rate of blood flow signals by CDFI,the detection rate of blood flow signals of grade 2 to 3,and BRAF gene mutation in the combined assessment of lymph node metastasis of thyroid cancer was 0.938,and the efficiency of combined assessment was better than that of the independent assessment of each index.Conclusion The ultrasound blood quantitative parameters combined with BRAF gene mutation has certain value in the evaluation of HT with thyroid cancer and lymph node metastasis,and the combined evaluation of each index is more effective.