OBJECTIVE To establish the high-performance liquid chromatography （HPLC） fingerprint of Sagina japonica ， and to establish a quantitative analysis of multi-components by single-marker （QAMS） method for simultaneous determination of six componen ts in S. japonica ， aiming to provide references for the quality control of this medicinal herb. METHODS HPLC method was used to establish the fingerprints of 12 batches （No. S1-S12） of S . japonica according to Similarity Evaluation System of Chromatographic Fingerprint of Traditional Chinese Medicine . The similarity evaluation and identification of common peaks were conducted， followed by cluster analysis （CA） and principal component analysis （PCA） for 12 batches of samples. Using vicenin-2 as internal reference， the contents of p-hydroxy cinnamic acid， apigenin-6-C-arabinoside-8-C-glucoside， isoorientin， vitexin and 20-hydroxyecdysone were determined by QAMS method. The results were then compared with those obtained by the external standard method. RESULTS The similarities of HPLC fingerprints for 12 batches of S . japonica ranged from 0.828-0.998. A total of 17 common peaks were calibrated， and 6 common peaks were identified. Specifically， peak 5 was identified as vicenin-2， peak 7 as p-hydroxycinnamic acid， peak 10 as apigenin-6-C-arabinoside-8-C-glucoside， peak 11 as isoorientin， peak 13 as vitexin， and peak 15 as 20-hydroxyecdysone. The results of CA showed that S1-S5， S7 and S9-S11 were clustered into one category， S6 was clustered into one category， and S8 and S12 were clustered into one category. The results of PCA revealed that the accumulative contribution rate of the four main components was 89.430%. The content ranges measured by QAMS method for p-hydroxy cinnamic acid， apigenin-6-C-arabinoside-8-C-glucoside， isoorientin， vitexin and 20-hydroxyecdysone were 0.017 4-0.269 4， 0.568 8-4.240 3， 0.503 2-5.040 3， 0.024 0-0.132 0 and 2.551 3-4.881 1 mg/g， respectively. There was no significant difference in the contents of components measured between QAMS method and the external standard method （ P ＞0.05）. CONCLUSIONS The established HPLC fingerprint and QAMS method can be used for quality evaluation and quality control of S . japonica.

AIM: To develop a novel gene-delivery therapeutic based on CRISPR/Cas9 genome editing technology capable of specifically targeting and knocking out the VEGFA gene, thereby achieving sustained suppression of VEGFA expression in retinal pigment epithelial(RPE)cells and providing a new strategy for gene therapy in retinal neovascular diseases.METHODS:Single guide RNAs targeting the human VEGFA gene for knockout were designed, and corresponding recombinant plasmids were constructed. A novel polymer(PTEE)was used to encapsulate the plasmids to prepare a PTEE-loaded anti-VEGFA plasmid(PLAP)gene delivery system. PTEE materials at concentrations of 0.1, 0.2, 0.4, 0.8, and 1.6 μg/μL were co-incubated with ARPE-19 cells, and the biocompatibility of PTEE was evaluated using the cell counting kit-8(CCK-8)assay. Recombinant plasmids expressing green fluorescent protein(GFP)were constructed. Lipofectamine 3000 and jetOPTIMUS®DNA transfection reagents were used as control groups, and PTEE nanomaterials were used as the experimental group to encapsulate the plasmids. When the cell confluence reached 80%, the formulations were transfected into ARPE-19 and 293T cells. GFP expression was observed under light microscopy, and the transfection efficiencies of each group were compared. ARPE-19 cells were induced under hypoxia, and PLAP was transfected into the cells. The expression level of VEGFA was detected by enzyme-linked immunosorbent assay(ELISA)to evaluate the efficacy of this novel gene delivery system.RESULTS: After co-incubation of ARPE-19 cells with different concentrations of PTEE for 24 h and 48 h, no significant effect on cell viability was observed in any group. The transfection efficiency of PLAP in ARPE-19 cells was higher than that in the Lipo3000 and jetOPTIMUS groups, with statistically significant differences(P<0.01). Hypoxia for 6 h significantly induced the upregulation of VEGFA mRNA expression in ARPE-19 cells, and under hypoxic conditions, the PTEE group exhibited a significant inhibitory effect on VEGFA expression(P<0.01).CONCLUSION:PLAP exhibits favorable biocompatibility and prominent VEGFA inhibitory effects in vitro, making it a potential candidate drug for gene therapy of retinal neovascular diseases.

Objective This study evaluated the diagnostic value of adding magnetic resonance imaging (MRI) to multimodal ultrasound for breast cancer, where multimodal ultrasound consisted of two-dimensional imaging and the strain ratio (SR) from elastography. Methods We enrolled 180 patients with breast lesions who were admitted to our hospital between August 2023 and August 2025. All patients underwent multimodal ultrasound and MRI examinations. Multimodal ultrasound included two-dimensional imaging and ultrasound elastography with strain ratio measurement. Breast cancer detection rates were assessed for each imaging method. The diagnostic performance of multimodal ultrasound alone, MRI alone, and the two methods combined was compared. Results Among the 180 patients, 82 (45.56%) had benign lesions and 98 (54.44%) had malignant lesions. The numbers of cases consistent with pathological diagnosis were 152 for multimodal ultrasound, 145 for MRI, and 160 for their combination. The sensitivity of the combined diagnosis was 95.92%, which exceeded that of multimodal ultrasound alone (84.69%) and MRI alone (79.59%). The negative predictive value of the combined diagnosis was 94.29%, which was significantly higher than that of MRI alone (77.01%, P<0.05). Conclusion Combining MRI with multimodal ultrasound based on SR and two-dimensional ultrasound significantly improves the differential diagnosis of breast cancer. This strategy compensates for the limitations of using a single modality and provides a more reliable imaging basis for clinical diagnosis and treatment decisions.

Objective:To identify the risk factors of venous thromboembolism (VTE) in postoperative renal transplantation recipients by Meta-analysis, and to provide evidence-based reference for clinical staff to develop early VTE prevention strategies.Methods:PubMed, Web of Science, Cochrane Library, Embase, China National Knowledge Infrastructure, VIP database, Wanfang database and Chinese Biomedical Literature Database were searched to collect the studies on the risk factors of postoperative VTE in kidney transplant recipients. The search period was from the establishment of the database to March 10, 2024. After literature screening, data extraction and quality evaluation were conducted independently by two researchers, Meta-analysis was performed using RevMan 5.3 software.Results:A total of 15 literatures with 20 influencing factors were included. Meta-analysis showed that age ( MD = 6.36, 95% CI 2.56-10.17, P<0.05), body mass index ( MD = 1.83, 95% CI 0.15-3.50, P<0.05), VTE history ( OR = 2.04, 95% CI 1.08-3.86, P<0.05), blood transfusion history ( OR = 3.77, 95% CI 2.43-5.83, P<0.05), glomerular filtration rate ( MD = -5.54, 95% CI -9.93 - -0.91, P<0.05), donor age ( MD = 3.18, 95% CI 1.10-5.25, P<0.05), combination of malignant tumor ( OR = 2.87, 95% CI 1.45-5.68, P<0.05), end-stage renal disease as polycystic kidney disease ( OR = 1.76, 95% CI 1.39-2.22, P<0.05), and interstitial nephritis ( OR = 1.60, 95% CI 1.06-2.40, P<0.05) were the influencing factors for postoperative VTE in renal transplant recipients. Conclusions:Clinical medical staff should actively identify high-risk groups for VTE after kidney transplantation by considering the 8 influencing factors determined by this study, and take targeted measures early to reduce the risk of postoperative VTE.

Objective:To explore the efficacy of letrozole combined with palbociclib in the treatment of hormone receptor +/human epidermal growth factor receptor-2 - (HR +/HER2 -) advanced breast cancer and its influence on serum thymidine kinase 1 (TK1) and proliferating cell nuclear antigen (Ki67) levels. Methods:A total of 82 patients with HR +/HER2 - advanced breast cancer, all admitted from Jan. 2022 to Jan. 2024 from Department of Oncology, Affiliated Hospital of Jining Medical University, were assigned to the control group ( n=41, letrozole) and the study group ( n=41, letrozole + palbociclib) according to the random number table. The therapeutic effect, tumor markers, immune function and serum TK1 and Ki67 levels were compared between the two groups. Results:The study group higher objective response rate and disease control rate than the other group [14.63% (6/41) and 85.37% (35/41) vs. 0.00% (0/41) and 58.54% (24/41) ] (Continuity correction χ2/χ2=4.50, 7.31, P<0.05). After treatment, carbohydrate antigen 153 (CA153), CA125 and carcinoembryonic antigen (CEA) in the study group were lower than the control group [ (12.17±3.19) U/mL, (23.57±3.35) U/mL and (19.51±4.13) ng/mL vs (24.37±5.25) U/mL, (35.16±5.08) U/mL, (34.28±5.72) ng/mL] ( t=12.72, 12.20, 13.41, P<0.05), serum TK1 and Ki67 levels were also lower [ (3.61±0.75) pmol/L and (7.89±1.16) ng/mL vs. (4.76±0.88) pmol/L and (10.85±1.94) ng/mL] ( t=6.37, 8.39, P<0.05). After treatment, CD3 +, CD4 +, CD4 +/CD8 + higher than the control group [ (48.64±5.88) %, (31.34±4.06) %, (1.22±0.27) vs. (43.16±6.09) %, (27.04±3.35) %, (0.87±0.35) ] ( t=4.15, 5.23, 5.07, P<0.05), while CD8 + was lower [ (25.73±4.11) % vs. (30.94±4.47) %] ( t = 5.49, P<0.05) . Conclusions:Letrozole combined with palbociclib is effective in the treatment of HR +/HER2 - advanced breast cancer and can reduce tumor markers and serum TK1 and Ki67 levels in patients.

Tissue-resident memory T cells(TRM)are a key immune memory cell type responsible for the recurrence of many skin diseases.TRM cells remain in the skin tissue following infection or injury and maintain immune memory by forming a memory reservoir.When the same or similar external stimuli are encountered again,TRM cells can be rapidly activated,releasing inflammatory cytokines and cytotoxic substances,leading to local tissue damage and/or disease recurrence.The formation and function of TRM cells are regulated by various factors,such as T cell receptor signals,cytokines,co-stimulatory molecules,etc.Furthermore,TRM cells may exhibit different phenotypes and functions in different types of skin diseases.Therefore,understanding the characteristics and regulatory mechanisms of TRM cells is of great significance for studying the mechanisms of skin disease recurrence as well as for developing therapeutic strategies targeting TRM cells.

Objective:To understand the pathogen distribution characteristics of acute lower respiratory tract infections (ALRTI) in hospitalized children in the Department of Respiratory Medicine, Capital Center for Chlidren's Health, Capital Medical University from 2019 to 2023, and to analyze their epidemiological features.Methods:A retrospective analysis was conducted, including 5 558 children aged 0-18 years who were hospitalized and diagnosed with ALRTI in the respiratory department of Capital Center for Chlidren's Health, Capital Medical University from December 8, 2019, to December 31, 2023. Nasopharyngeal swabs, sputum, and bronchoalveolar lavage fluid specimens were collected for pathogen detection.Results:Among the 5 558 ALRTI hospitalized children, the positive detection rate of pathogens was 81.81% (4 547/5 558). In bronchoalveolar lavage fluid specimens, Mycoplasma pneumoniae had the highest detection rate (33.52%), while Streptococcus pneumoniae had the highest detection rate (24.38%) in upper respiratory tract specimens. Mycoplasma pneumoniae (54.21%, χ2=1 034.58, P<0.001) and adenovirus (5.09%, χ2=10.84, P=0.013) were most common in school-age children, human rhinovirus was more common in toddlers (26.42%, χ2=122.36, P<0.001), and respiratory syncytial virus had the highest infection rate in infants (28.61%, χ2=326.47, P<0.001). A total of 52.06% of the children had mixed infections (2 367/4 547), with the highest proportion of mixed infections in infants (62.54%). During the post- non-pharmaceutical interventions (NPIs) period, the proportions of Mycoplasma pneumoniae (69.88% vs. 58.16%), human rhinovirus (30.37% vs. 14.02%), adenovirus (51.16% vs. 6.45%), influenza A virus (36.46% vs. 16.67%), and mixed infections (33.33% vs. 19.04%) in school-age children were significantly higher than during the NPIs period. In contrast, the proportions of respiratory syncytial virus (33.19% vs. 19.42%), adenovirus (11.29% vs. 4.65%), and mixed infections (17.75% vs. 10.89%) in infants were significantly lower than during the NPIs period. Conclusion:NPIs have changed the detection rate and epidemiology of respiratory pathogens in hospitalized children. After the end of NPIs, outbreaks of some pathogens may occur, so it is necessary to strengthen rapid pathogen detection, population surveillance, and health education.

Viruses constitute a significant group of pathogens that have caused numerous fatalities and substantial economic losses in recent years, particularly with the emergence of coronaviruses. While the impact of SARS-CoV-2 appears to be diminishing in daily life, only a limited number of drugs have received approval or emergency use authorization for its treatment. Given the high mutation rate of viral genomes, host-directed agents (HDAs) have emerged as a preferred choice due to their broad applicability and lasting effectiveness. In contrast to direct-acting antivirals (DAAs), HDAs offer several advantages, including broad-spectrum antiviral activities, potential efficacy against future emerging viruses, and a lower likelihood of inducing drug resistance. In our review article, we have synthesized known host-directed antiviral targets that span diverse cellular pathways and mechanisms, shedding light on the intricate interplay between host cells and viruses. Additionally, we have provided a brief overview of the development of HDAs based on these targets. We aim for this comprehensive analysis to offer valuable perspectives and insights that can guide future antiviral research and drug development efforts.

Coronavirus-related diseases pose a significant challenge to the global health system. Given the diversity of coronaviruses and the unpredictable nature of disease outbreaks, the traditional "one bug, one drug" paradigm struggles to address the growing number of emerging crises. Therefore, there is an urgent need for therapeutic agents with broad-spectrum anti-coronavirus activity. Here, we provide evidence that ATV006, an anti-SARS-CoV-2 nucleoside analog targeting RNA-dependent RNA polymerase (RdRp), has broad antiviral activity against human and animal coronaviruses. Using mouse hepatitis virus (MHV) and human coronavirus NL63 (HCoV-NL63) as a model, we show that ATV006 has potent prophylactic and therapeutic activity against murine coronavirus infection in vivo. Remarkably, ATV006 successfully inhibits viral replication in mice even when administered 96 h after infection. Due to its oral bioavailability and potency against multiple coronaviruses, ATV006 has the potential to become a useful antiviral agent against SARS-CoV-2 and other circulating and emerging coronaviruses in humans and animals.

The regulatory processes in developmental biology research are significantly influenced by long non-coding RNAs (lncRNAs). However, the dynamics of lncRNA expression during human tooth development remain poorly understood. In this research, we examined the lncRNAs present in the dental epithelium (DE) and dental mesenchyme (DM) at the late bud, cap, and early bell stages of human fetal tooth development through bulk RNA sequencing. Developmental regulators co-expressed with neighboring lncRNAs were significantly enriched in odontogenesis. Specific lncRNAs expressed in the DE and DM, such as PANCR, MIR205HG, DLX6-AS1, and DNM3OS, were identified through a combination of bulk RNA sequencing and single-cell analysis. Further subcluster analysis revealed lncRNAs specifically expressed in important regions of the tooth germ, such as the inner enamel epithelium and coronal dental papilla (CDP). Functionally, we demonstrated that CDP-specific DLX6-AS1 enhanced odontoblastic differentiation in human tooth germ mesenchymal cells and dental pulp stem cells. These findings suggest that lncRNAs could serve as valuable cell markers for tooth development and potential therapeutic targets for tooth regeneration.
Humans ; RNA, Long Noncoding/metabolism* ; Odontogenesis/genetics* ; Tooth Germ/embryology* ; Cell Differentiation ; Gene Expression Regulation, Developmental ; Mesoderm/metabolism* ; Tooth/embryology* ; Gene Expression Profiling ; Sequence Analysis, RNA ; Dental Pulp/cytology*