OBJECTIVE To study the effects of bergapten in the treatment of liver fibrosis and its mechanism based on serum metabolomics. METHODS Forty mice were divided into normal control group （0.5% carboxymethyl cellulose sodium solution）， model group （0.5% carboxymethyl cellulose sodium solution）， and BP low-dose and high-dose groups （50， 100 mg/kg）， with 10 mice in each group. Except for the normal control group， the other three groups were all treated with carbon tetrachloride to induce liver fibrosis model； they were given relevant medicine/solution intragastrically， once a day， for consecutive 8 weeks. After the last medication， the levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum were detected， and liver pathological changes were observed； the expressions of α-smooth muscle actin （α-SMA） and Collagen Ⅰ were detected in liver tissue； the serum of the mice was collected for metabolomics analysis. RESULTS Compared with the model group， serum levels of ALT and AST and protein expressions of α-SMA and Collagen Ⅰ in liver tissue were decreased significantly in BP high-dose and low-dose groups （P＜0.05）， while liver fibrosis was improved significantly. Meanwhile， metabolomics analyses showed that there were a total of 175 serum differential metabolites in the BP high-dose group and model group， of which 18 substances were upregulated and 157 substances were downregulated； the main metabolic pathways involved in bergapten intervention were pyrimidine metabolism， butanoate metabolism， fatty acid synthesis， tyrosine metabolism， β-alanine metabolism， nicotinic acid and nicotinamide metabolism， glutathione metabolism， etc. CONCLUSIONS BP is effective in the treatment of liver fibrosis by regulating pyrimidine metabolism， butanoate metabolism， glutathione metabolism and so on in rats with liver fibrosis.

Objective To explore the characteristics of echolalic speech of children with moderate to severe autism spectrum disor-der(ASD)aged three to eight. Methods From January to March,2024,15 minutes rehabilitation course video of five children with moderate to severe ASD aged three to eight were collected in institutional and family settings,respectively.Language sample analy-sis and self-compiled coding were used to analyze the echo speech sample content,structure,and collect their oral language ability.The correlation between the echo speech and case oral language ability was analyzed. Results A total of 235 echoic speech-related speech rounds were recorded.Among them,the ratio of immediacy to de-layed echo speech was 7∶1;one echo speech was found in every two to three speech responses,with average sen-tence length of 2.97 and average word length of 2.23.The frequency of verbs and nouns was high,accounting for 45.03%and 38.48%,respectively;and there were many declarative words and sentences,accounting for 72.58%on average.The type of echo speech structure was the most frequently at the end of the reconstructed discourse,accounting for 29.98%on average.The type,frequency,sentence length,sentence pattern and structure of echoic speech were not significantly different in institutional and family settings.The language ability scores of imita-tion sentence length,word understanding and naming were significantly correlated with the number,frequency,sentence length and sentence type frequency of echoic speech(r>0.881,P<0.01). Conclusion Echoic speech activity in children with moderate to severe ASD aged three to eight shows transsituational stability,and is related with their understanding language and expressive language ability;characterized as instant echo speech,short sentence,the advantage of real words,declarative sentences,often repeat the end of the origi-nal sentence.

Objective:To investigate the impact of exogenous lactate on the replication of dengue virus type 2 (DENV-2) in Raw264.7 cells, mouse bone marrow-derived macrophages (BMDMs) and THP-1 cells and explore its association with cell activation.Methods:BMDMs from BALB/c mouse bone marrow were prepared and evaluated by flow cytometry to detect the proportion of F4/80 + CD11b + cells. Glucose transporter type 1 (GLUT1), hexokinase 2 (HK2), and monocarboxylate transporters 4 (MCT4) expression at mRNA level in BMDMs at different time points after DENV-2 infection were measured by qRT-PCR. The content of lactate in the culture supernatants was quantified via colorimetric assay. CCK-8 assay was used to evaluate the impacts of different concentrations of lactate on the viability of Raw264.7 cells, BMDMs, and THP-1 cells. qRT-PCR was used to detect the expression of DENV-2 E gene, TGF-β, CD86, retinoic acid-inducible gene Ⅰ (RIG-Ⅰ), IFN-β, interferon-stimulated gene 15 (ISG15), and ISG56 at mRNA level in cells infected with DENV-2 at different MOIs in the presence of different concentrations of lactate. Meanwhile, flow cytometry was used to analyze the expression of CD86 and CD206. Results:The percentage of BMDMs was (87.53±1.66)%. GLUT1 expression at mRNA level exhibited a decrease in BMDMs at 24 h after DENV-2 (MOI=3) infection following a transient increase at 12 h ( P<0.05), while HK2 expression at mRNA level was higher that than in blank control and inactivated DENV-2 infection groups at 12, 24, and 36 h ( P<0.01). Besides, there was an increase in both MCT4 mRNA level and the content of lactate in culture supernatants at 24 h after DENV-2 (MOI=1.5) infection ( P<0.05). The viability of the three types of cells remained above 80% when the concentration of lactate was 31.25 mmol/L. Lactate at the concentration of 35 mmol/L increased the expression of the DENV E gene at mRNA level in DENV-2-infected BMDMs at MOI=1 or MOI=2 ( P<0.05). Besides, it promoted the expression of DENV E gene at mRNA level in Raw264.7 and THP-1 cells ( P<0.001) as well as the expression of CD163, TGF-β, RIG-Ⅰ, IFN-β, ISG15 and ISG56 at mRNA level in BMDMs at MOI=1.5, but inhibited the expression of CD86 at mRNA level in BMDMs ( P<0.05). It also up-regulated CD206 protein expression ( P<0.01) and down-regulated CD86 protein expression ( P>0.05) in BMDMs. Conclusions:Exogenous lactate enhances DENV-2 replication in both human- and murine-derived macrophages and that might correlate with M2 macrophage polarization.

Objective:To investigate the genomic manifestation and pathogenesis of osteosarcoma with different relapse pattens, which were respectively initially presented with bone metastasis or pulmonary metastasis.Methods:From May 1, 2021 to October 1, 2021, 38 fresh tumor specimens and some paraffin-embedded specimens of high-grade osteosarcoma were collected in Peking University People's Hospital, including 29 males and 9 females, aged 19.6±2.2 years (range, 6-61 years). Among the 38 cases, 12 cases had initial bone metastasis (group A) and 26 cases had initial lung metastasis (group B), of which 15 cases (40%, 15/38) had paired specimens of primary and metastatic lesions. Based on Illumina NovaSeq 6000, we analyzed whole-exome sequencing (WES) as well as transcriptome for osteosarcoma with paired samples in different relapse patterns. During all their treatment courses, we also collected their paired samples to reveal these tumors' evolution. We sought to redefine disease subclassifications for osteosarcoma based on genetic alterations and correlate these genetic profiles with clinical treatment courses to elucidate potential evolving cladograms.Results:We found that osteosarcoma in group A mainly carried single-nucleotide variations (83%, 10/12), displaying higher tumor mutation burden [4.9 (2.8, 12.0) & 2.4 (1.4, 4.5), P=0.010] and neoantigen load [743.0 (316.5, 1,034.5) & 128.5 (49.0, 200.5), P=0.003], while those in group B mainly exhibit structural variants (58%, 15/26). The mutation spectrum showed that there was a significant difference in age-related gene imprinting 1 between the bone metastasis group and the lung metastasis group ( P=0.005). Samples were randomly selected from group A (3 patients) to investigate immunologic landscape by multiplex immunohistochemistry, from which we noticed tertiary lymphatic structure from one patient from group A. High conservation of reported genetic sequencing over time was found in their evolving cladograms. Conclusion:Osteosarcoma with mainly single-nucleotide variations other than structural variants might exhibit biological behavior predisposing toward bone metastases with older in age as well as better immunogenicity in tumor microenvironment.

Abstract OBJECTIVE To investigate the effect and mechanism of flavonoids from Sophora flavescens Ait. on testicular tissue damage in male mice induced by local heat stress in the scrotum. METHODS TCMSP database was used to screen the targets of flavonoids in Sophora flavescens Ait., and the bioinformatics analysis was performed on the target. The mouse model of scrotal heat stress was used and the flavonoids of Sophora flavescens Ait. was used for intervention. The sperm density and sperm aberration rate of mice in each group were measured, and the morphological changes of testicular tissue were observed. Tumor necrosis factor-α (TNF-α) and interleukin 17(IL-17) mRNA and protein levels in testicalar were detected of by q-PCR and Western blotting. Na+-K+-ATPase, Mg2+-ATPase, Ca2+-ATPase, lactate dehydrogenase(LDH), sorbitol dehydrogenase(SDH) activity, malondialdehyde (MDA), NO, TNF-α level and the content of testosterone in serum were detected in tissue homogenate. RESULTS Heat stress could lead to the decrease of sperm density and increase of aberrant sperm, the obvious thinning of testicular spermatogenic epithelium, the decrease of cell level and quantity, the significant decrease of ATPase, LDH, SDH activities, and the increase of MDA, NO content, TNF-α and IL-17 expression in testicular tissue. After the intervention with 250, 500 mg·kg-1·d-1 flavonoids of Sophora flavescens Ait., the quality of sperm and the damage of testicular tissue morphology were improved. The level of TNF-α and IL-17 in serum and testicular tissue were decreased, and the activity of ATPase, SDH and the level of testosterone were increased. CONCLUSION The mechanism of flavonoids of Sophora flavescens Ait. is through inhibiting the inflammatory factor TNF-α and IL-17 levels, improve the anti-lipid peroxidation ability and inhibite the role of NO, enhance the activity of energy enzymes in spermatogenesis, improve the level of serum testosterone, and improve the reproductive disorders caused by heat stress.

Objective:To explore the effect of venetoclax-based therapy on relapsed/refractory multiple myeloma (MM) patients harboring t(11;14).Methods:The data of a relapsed/refractory MM patient harboring t(11;14) treated with venetoclax-based regimen admitted to Shanghai Changzheng Hospital in June 2019 was retrospectively analyzed and the literatures were reviewed.Results:The relapsed/refractory MM patient harboring t(11;14) had progression of disease after 3 lines of therapies, and then was treated with the selective bcl-2 inhibitor venetoclax combined with daratumumab and dexamethasone. As a result, the patient achieved partial remission and better hemogram recovery. The Eastern Cooperative Oncology Group (ECOG) score of physical status decreased from 3 to 1, and the quality of life was improved significantly.Conclusions:The relapsed/refractory MM patients harboring t(11;14) could benefit from venetoclax-based therapy. In the future, the safety, sensitivity and other performances of venetoclax in the treatment of MM should be further explored.

Liquid biopsy can non-invasively reveal the status of tumors and its prognosis in vivo, which is hotspots and difficulties of research in current era. Among them, an emerging marker called circulating tumor RNA (ctRNA) reflects the genetic information of tumor origin and provides a powerful basis for early diagnosis, targeted drug monitoring and prognosis prediction. At this stage, several ctRNA kits have been approved for clinical use. For example, microRNA (miRNA) can be used for aided diagnosis of liver cancer. And many transformation applications of promising ctRNA have been vigorously carried out. Despite the facts that there are still many clinical challenges of ctRNA detection technology to be solved effectively, ctRNA, as a new member of the liquid biopsy technology, has shown remarkable clinical value. Along with the mechanism becoming gradually clear, ctRNA will be a reliable diagnostic tool with the increasing clinical requirements for facilitating the tumor management.

Objective: To investigate the bacterial species, characteristics and differences of oral bacteria flora of saliva in the longevous elderly between in Bama county and in Debao county in Guangxi, in order to explore the relationship between longevity and oral salivary bacteria flora in the elderly. Methods: The saliva was taken from the longevous elderly in Bama county(BM group)and people aged over 60 years in Debao county(BS group)separately, and the total DNA was extracted.The 16S rDNA-V4 region was amplified by PCR and analyzed by sequencing.The main species and diversity of bacterial colonies were recorded for difference analysis. Results: A total of 14 saliva samples were collected from 7 cases in BM group and 7 cases in BS group.A total of 369 OTUs were generated by cluster analysis of 14 samples.At the genus level, the dominant salivary bacteria flora were Ctinomyces, Capnocytophaga, Chryseobacterium, Fusobacterium, Haemophilus, Lactobacillus, Leptotrichia, Neisseria, Porphyromonas, Prevotella, Rothia, Streptococcus, Veillonella in both BM group and BS group.The OTU PCA analysis showed that some evidence for indeterminate differences was found, but statistically significant differences did not exist in the dominant components of oral flora between the two groups(P>0.05). Also, the same tendency toward the diversity(P>0.05)was presented.Similarly, the species annotation analysis and the heat map showed that there were no significant differences(P>0.05)in oral salivary flora composition between the two groups.Lactobacillu was always the prevailing flora in the Phylume, Class, Order, Family and Genus, but the abundance ratio was different between the two groups as following: Lactobacillus abundance in salivary bacteria flora was higher in BM Group than in the BS group, while Mycoplasma abundance was lower in BM Group than in the BS group(P<0.05). Conclusions The dominant salivary bacteria flora is Lactobacillus in both BM and BS group, while, the abundance of Lactobacillus is higher in the BM group than in the BS group, which indicates that the longevity of population in Bama county may be related to Lactobacillus.

Liquid biopsy can non-invasively reveal the status of tumors in vivo, and provides a powerful basis for early diagnosis, personalized treatment monitoring and prognosis prediction. According to the type of tumor-associated material, liquid biopsy covers circulating tumor cells (CTCs), circulating tumor DNA (ctDNA), extracellular vesicles (EVs) and circulating tumor RNA (ctRNA) etc. At present, several liquid biopsy products havebeen approved for clinical use, while many transformation studies have been vigorously carried out. However, there are still many clinical challenges to be solved effectively. Despite the standardization of detection and quality management system of liquid biopsy are lagged with the rapid development of technology, liquid biopsy, as a highly promising detection technology, will become a reliable diagnostic tool with the increasing clinical requirements for facilitating the tumor management.

Objective:To summarize the clinical characteristics and prognosis of 51 patients with Waldenstr?m’s macroglobulinemia (WM) and evaluate the efficacy and adverse reactions of ibrutinib in the treatment of WM.Methods:We carried out a single-center retrospective study, including 51 patients with WM of our single center from November 2008 to October 2019.Results:The median age at diagnosis was 65 years with a male-to-female ratio of 2.64∶1. There were 9 (18%) , 21 (41%) , and 21 (41%) ISSWM stage low-, intermediate- and high-risk patients identified, respectively. A total of 27 (73%) patients harbored MYD88 L265P mutation. The median follow-up time was 38.6 (0.3-120.0) months, the median progression free survival was 46.4 months, and the median overall survival was not reached. The overall remission and major remission rates of patients who received ibrutinib were 87% and 80%, respectively. The median time to achieve at least partial remission of patients treated with ibrutinib was 8 weeks, which was earlier than those treated with other drugs ( P<0.05) . Conclusion:WM is often seen in elderly men. MYD88 L265P had a high frequency in WM. The findings of our study validate the efficacy of ibrutinib monotherapy. Even in patients with advanced age and at high risk of ISSWM, the overall remission rate and major remission rate are high. Ibrutinib is a safe and effective therapy because of its rapid onset and rare serious adverse reactions.