Aim To investigate the effects of bone-forming protein BMP9 on aerobic glycolysis,migration and invasion ability in triple-negative breast cancer MDA-MB-231 cells and the underlying mechanisms.Methods The experimental group infected MDA-MB-231 cells with human BMP9 recombinant adenovirus(AdBMP9),while the control group infected cells with empty GFP adenovirus.Lactate,glucose and ATP as-say kits were used to detect glucose uptake,lactate and ATP production.The correlation between BMP9 and key glycolytic enzyme genes in pancarcinoma was ana-lyzed using GEPIA2 database.The mRNA expression levels of GLUT1,HK2,PKM2 and LDHA in MDA-MB-231 cells after overexpression of BMP9 were detec-ted by qRT-PCR.Potential targets of BMP9 inhibiting MDA-MB-231 aerobic glycolysis were analyzed in STRING database.The expression levels of HIF-1αand downstream protein were detected by Western blot.The changes of cell migration and invasion ability after different treatments were evaluated by the scratch heal-ing assay and Transwell assay.Results Compared with the control group,BMP9 down-regulated glucose uptake,lactate production,ATP level(P＜0.01),and inhibited HIF-1α and its downstream protein ex-pression in MDA-MB-231 cells.Overexpression of HIF-1α in rescue experiment reversed the inhibitory effect of BMP9 on aerobic glycolysis,migration and in-vasion of MDA-MB-231 breast cancer cells.Conclu-sion BMP9 down-regulates HIF-1α to inhibit the aer-obic glycolysis and migration and invasion ability of MDA-MB-231 breast cancer cells.

Aim To investigate the effects of autophagy regula-ted by LncRNA SNHG3 on the proliferation,migration,invasion and EMT of human breast cancer cell line MCF-7.Methods The expression of SNHG3 in breast cancer and breast cancer cell line MCF-7 was analyzed by bioinformatics and real-time fluores-cent quantitative PCR(RT-qPCR);RNAi technology was used to construct MCF-7 recombinant cell lines with knockdown SNHG3(siSNHG3)and control(siNC),and Western blot and cellular immunofluorescence were applied to detect autophagy markers;autophagosome lysosomal fusion inhibitor BafA1 or ear-ly autophagosome formation inhibitor 3-MA was employed to treat MCF-7 cells with or without SNHG3 knockdown,Western blot was used to detect the expression of LC3-Ⅱ or p62,and the effect on autophagic vesicle formation or autophagic degradation was observed;clone formation experiment,CCK8 experiment,wound healing experiment,and Transwell experiment were used to detect the effects of siSNHG3 combined or not combined with BafA1 or 3-MA on the proliferation,lateral migration,longitudi-nal migration,and invasion of MCF-7 cells.Western blot was used to detect its effect on the EMT of MCF-7 cells.Results Bioinformatics analysis and RT-qPCR confirmed that SNHG3 was highly expressed in breast cancer and breast cancer cell line MCF-7;Western blot and cellular immunofluorescence confirmed that knockdown of SNHG3 could activate autophagy in breast cancer;the clone formation,CCK-8,wound healing,and Tran-swell experiment confirmed that downregulation of SNHG3 ex-pression could inhibit tumor proliferation,migration,and inva-sion by activating autophagy;Western blot confirmed that SNHG3 promoted EMT process of breast cancer through negative regulation of autophagy.Conclusions SNHG3 is abnormally overexpressed in breast cancer and negatively regulates autoph-agy,and can enhance the proliferation,migration,invasion and EMT process of breast cancer cells through negatively regulating autophagy,suggesting that SNHG3 is a potential target for diag-nosis and treatment of breast cancer.

In recent years,the number of Chest Pain Centers in China has increased rapidly.As of December 2023,there were 5 725 registered units of Chest Pain Centers nationwide,2 660 of which had passed the acceptance,and a total of 14.67 million patients with acute chest pain were treated.The Chest Pain Centers treated more than 3.18 million patients with acute chest pain in 2023,including more than 690 000 patients with acute myocardial infarction,and achieved positive results:significantly increasing the proportion of reperfusion therapy for ST-segment elevation myocardial infarction patients,ensuring the treatment of myocardial infarction patients in a short time,shortening the length of patient hospitalization,and reducing the hospitalization costs and mortality of patients.Through the construction of chest pain center,the treatment process was significantly optimized and the prognosis of patients was improved.At present,the construction and quality control of Chest Pain Centers still need to be further promoted.In some areas,the construction proportion of Chest Pain Centers and the level of diagnosis and treatment of chest pain need to be improved,patient delay is still an important challenge for the treatment of acute chest pain in China.

The molecular epidemiological characteristics of 1,4,[5],12:i:-Salmonella in Jiangsu Province were analyzed through whole genome sequencing(WGS).The distribution characteristics of related genes were obtained on the basis of anno-tated drug-resistant genes and plasmid types in the whole genome.Analysis of the molecular epidemiological characteristics of strains with cgMLST revealed possible modes of transmission of quinolone resistance in 1,4,[5],12:i:-Salmonella.Eleven cat-egories of antibiotic resistance genes(ARGs)were annotated among the fluoroquinolone-resistant strains.The detection rate of aminoglycoside ARGs was highest(100％).Twelve quinolone-resistant strains(92.3％)carried the IncHI2/IncHI2A plasmid type.PMQR gene analysis of various strains indicated that the strains from the United States and Europe carried six types of PMQR genes,and the detection rate of qnrB19 was highest.The Jiangsu strains carried three PMQR gene types,and the de-tection rate of aac(6')-Ib-cr was highest(11.84％).Analysis of cgMLST loci from different countries/regions revealed three main epidemic clusters.Some isolates from Jiangsu province might have the same evolutionary origin as some isolates from Eu-rope and the United States,and national/regional differences were observed in the PMQR gene carriage level.

Coronary restenosis is an important cause of poor long-term prognosis in patients with coronary heart disease. Here, we show that lysine methyltransferase SMYD2 expression in the nucleus is significantly elevated in serum- and PDGF-BB-induced vascular smooth muscle cells (VSMCs), and in tissues of carotid artery injury-induced neointimal hyperplasia. Smyd2 overexpression in VSMCs (Smyd2-vTg) facilitates, but treatment with its specific inhibitor LLY-507 or SMYD2 knockdown significantly inhibits VSMC phenotypic switching and carotid artery injury-induced neointima formation in mice. Transcriptome sequencing revealed that SMYD2 knockdown represses the expression of serum response factor (SRF) target genes and that SRF overexpression largely reverses the inhibitory effect of SMYD2 knockdown on VSMC proliferation. HDAC3 directly interacts with and deacetylates SRF, which enhances SRF transcriptional activity in VSMCs. Moreover, SMYD2 promotes HDAC3 expression via tri-methylation of H3K36 at its promoter. RGFP966, a specific inhibitor of HDAC3, not only counteracts the pro-proliferation effect of SMYD2 overexpression on VSMCs, but also inhibits carotid artery injury-induced neointima formation in mice. HDAC3 partially abolishes the inhibitory effect of SMYD2 knockdown on VSMC proliferation in a deacetylase activity-dependent manner. Our results reveal that the SMYD2-HDAC3-SRF axis constitutes a novel and critical epigenetic mechanism that regulates VSMC phenotypic switching and neointimal hyperplasia.

Objective:To evaluate the effect of electroacupuncture on P2X4R-p38 mitogen-activated protein kinase (p38 MAPK)-brain-derived neurotrophic factor (BDNF) signaling pathway in trigeminal ganglion of rats with trigeminal neuralgia.Methods:Thirty-six clean-grade healthy adult male Sprague-Dawley rats, weighing 190-230 g, aged 2-3 months, were divided into 3 groups ( n=12 each) using a random number table method: sham operation group (S group), trigeminal neuralgia group (TN group), and electroacupuncture group (E group). The model was developed by chronic constriction of the infraorbital nerve in anesthetized animals. The infraorbital nerve was only exposed without ligation in group S. Rats received electroacupuncture stimulation at the Baihui and Xiaguan acupoints on the affected side for 20 min after developing the model, with a frequency of 80 Hz, twice a day, for 14 consecutive days in E group. Facial mechanical pain threshold (FMT) was measured at 1 day before developing the model and 3, 7, 14, 21 and 28 days after developing the model. The rats were sacrificed after the last behavioral testing, and the trigeminal ganglia were taken for examination of histopathological changes of trigeminal ganglion (by HE staining) and for determination of the expression of P2X4R, p38 MAPK, phosphorylated p38 MAPK (p-p38 MAPK) and BDNF (by Western blot) and contents of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and IL-6 (by enzyme-linked immunosorbent assay). Results:Compared with group S, the FMT was significantly decreased at each time point after developing the model, the expression of P2X4R, p-p38 MAPK and BDNF in trigeminal ganglion was up-regulated, and the contents of TNF-α, IL-1β and IL-6 were increased ( P<0.05), the pathological changes of the trigeminal ganglion were obvious in group TN. Compared with group TN, the FMT was significantly increased at each time point after developing the model, and the expression of P2X4R, p-p38 MAPK and BDNF in trigeminal ganglion was down-regulated, and the contents of TNF-α, IL-1β and IL-6 were decreased ( P<0.05), and the pathological changes of the trigeminal ganglion were significantly attenuated in group E. Conclusions:The mechanism by which electroacupuncture alleviates trigeminal neuralgia may be related to inhibiting the activity of P2X4R-p38MAPK-BDNF signaling pathway and reducing neuroinflammation in rats.

Objective: To investigate the infection sources and the transmission chains of three outbreaks caused by 2019-nCoV Omicron variant possibly spread through cross-border logistics in Beijing. Methods: Epidemiological investigation and big data were used to identify the exposure points of the cases. Close contacts were traced from the exposure points, and the cases' and environmental samples were collected for nucleic acid tests. Positive samples were analyzed by gene sequencing. Results: The Omicron variant causing 3 outbreaks in Beijing from January to April, 2022 belonged to BA.1, BA.1.1 and BA.2. The outbreaks lasted for 8, 12 and 8 days respectively, and 6, 42 and 32 cases infected with 2019-nCoV were reported respectively. International mail might be the infection source for 1 outbreak, and imported clothes might be the infection sources for another 2 outbreaks. The interval between the shipment start time of the imported goods and the infection time of the index case was 3-4 days. The mean incubation period (Q₁, Q₃) was 3 (2,4) days and the mean serial interval (Q₁, Q₃) was 3 (2,4)days. Conclusions: The 3 outbreaks highlighted the risk of infection by Omicron variant from international logistics-related imported goods at normal temperature. Omicron variant has stronger transmissibility, indicating that rapid epidemiological investigation and strict management are needed.
Humans ; Beijing ; SARS-CoV-2 ; COVID-19 ; Disease Outbreaks ; China/epidemiology*

OBJECTIVE: To investigate the mechanism of shikonin-induced death of human hepatocellular carcinoma SMMC-7721 cells. METHODS: Cultured SMMC-7721 cells and normal hepatocytes (L-02 cells) were treated with 4, 8, or 16 μmol/L shikonin, and the changes in cell viability was assessed using MTT assay. The levels of ATP and lactic acid in the cell cultures were detected using commercial kits. Co-immunoprecipitation and immunofluorescence staining were used to determine the relationship among pyruvate kinase M2 (PKM2), prolyl hydroxylase 3 (PHD3), and hypoxia-inducible factor-1α (HIF-1α). The expressions of PHD3, PKM2, HIF-1α, Bax, cleaved caspase-3, and Bcl-2 in SMMC-7721 cells were detected with Western blotting, and cell apoptosis was analyzed with annexin V-FITC/PI staining. The effects of RNA interference of PKM2 on PHD3 and HIF-1α expressions in SMMC-7721 cells were detected using Western blotting. RESULTS: The IC₅₀ of shikonin against SMMC-7721 and L-02 cells was 8.041 μmol/L and 31.75 μmol/L, respectively. Treatment with shikonin significantly inhibited the protein expressions of PKM2, HIF-1α and PHD3 and nuclear translocation of PKM2 and HIF-1α in SMMC-7721 cells. Coimmunoprecipitation and immunofluorescence staining confirmed that shikonin inhibited the formation of PKM2/PHD3/HIF-1α complex and significantly reduced the contents of lactic acid and ATP in SMMC-7721 cells (P < 0.05). The expressions of PHD3 and HIF-1α decreased significantly after PKM2 knockdown (P < 0.05). Shikonin treatment significantly increased the apoptosis rate, enhanced the expressions of Bax and cleaved caspase-3, and decreased Bcl-2 expression in SMMC-7721 cells (P < 0.05). CONCLUSIONS Shikonin induces apoptosis of SMMC-7721 cells possibly by inhibiting aerobic glycolysis through the PKM2/PHD3/HIF-1α signaling pathway to cause energy supply dysfunction in the cells.
Humans ; Prolyl Hydroxylases ; Carcinoma, Hepatocellular ; Caspase 3 ; bcl-2-Associated X Protein ; Liver Neoplasms ; Signal Transduction ; Apoptosis ; Adenosine Triphosphate

Objective:To explore the difference of lymphocyte subsets in peripheral blood(PB)between aplastic anemia(AA)and hypoplastic myelodysplastic syndrome(hypo-MDS)patients,meanwhile to compare the clinical parameters obtained from PB and bone marrow(BM).Methods:The lymphocyte subsets in hypo-MDS(n=25)and AA(n=33)patients were investigated by flow cytometry.Meanwhile,the differences in PB cell counts,biochemical indicators,BM cell counts and abnormal chromosomes between the two groups were analyzed.Results:The percentage of CD8+T cells in A A group was significantly higher than that in hypo-MDS group(P=0.001),while the percentage of CD4+T cells and the CD4+/CD8+ratio in AA group were obviously lower than those in hypo-MDS group(P=0.015 and0.001,respectively).Furthermore,the proportion of CD4+andCD8+activated T(TA)cells,and memory Tregs in AA group was distinctly lower than those in hypo-MDS group(P=0.043,0.015 and 0.024,respectively).Nevertheless,the percentage of CD8+naive T(TN)cells in AA patients was remarkably higher(P=0.044).And hypo-MDS patients had declined lymphocyte counts(P=0.025),increased levels of total bilirubin(TBil),lactate dehydrogenase(LDH),vitamin B12 and proportion of BM blasts than AA patients(P=0.019,0.023,0.027 and 0.045,respectively).Conclusion:In this study it was confirmed that the percentages of CD4+and CD8+TA cells,memory Tregs and CD8+TN cells were significantly different between AA and hypo-MDS patients,which provide an essential basis for the identification of these two diseases.

Objective To analyze the epidemiological characteristics of norovirus infectious diarrhea in the active monitoring of foodborne diseases in Jiangsu Province from 2015 to 2020, so as to provide reference for the prevention and control of foodborne diseases caused by norovirus. Methods Norovirus positive diarrhea cases were collected from sentinel hospitals in 13 districts and cities of Jiangsu Province, and their epidemiological and clinical characteristics were analyzed. Results Atotal of 3 620 norovirus positive cases were detected and isolated from 61 489 samples. The main serotype was GII (71.97%), the onset season was winter and spring, and the onset age was 1-3 years old and 14-34 years old. There was no significant difference in norovirus positive rate between different sexes, and the main symptom was diarrhea (incidence rate was 92.10%), Meat and meat products (20.20%) were the main types of suspected exposed foods. Conclusion Norovirus infection has obvious seasonal characteristics, and the population is generally susceptible. It is high in children and young people, and meat food was the main suspicious exposure food. We should continue to improve the ability of active monitoring, identification, early warning and control of foodborne diseases, so as to reduce the occurrence of foodborne diseases.