Objective : To explore the mechanism of isorhamnetin (Iso) in the treatment of oral squamous cell carcinoma (OSCC) using network pharmacology and molecular docking methods and to verify it in vitro. Methods : The key targets were obtained by constructing the PPI protein interaction network based on the common intersection targets of Iso-OSCC. At the same time, gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) were used to analyze the related signaling pathways of the intersection targets. Iso and core targets were also analyzed through molecular docking and visualization. Colony formation assay and Transwell assay were used to identify the effect of Iso on the proliferation and invasion of Cal-27 cells. Western blot was used to analyze the regulatory effects of different concentrations of Iso on estrogen receptor-1 (ESR1), phosphoinositide-3-kinase regulatory subunit-1 (PIK3R1), Src tyrosine kinase (SRC), and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway proteins. Results: A total of 269 potential intersection targets of Iso-regulated OSCC were obtained. According to the degree obtained by topological analysis, PIK3R1, AKT1, SRC, ESR1, and other core targets were screened out. KEGG analysis showed that 165 signaling pathways were enriched in the intersection targets of Iso-OSCC, among which the PI3K/AKT signaling pathway played an important role in the treatment of OSCC with Iso. Molecular docking results showed that the absolute value of binding energy between target proteins PIK3R1, AKT1, SRC, ESR1, and Iso was high. After Cal-27 cells were treated with Iso, the number of cell colony formations, the number of transmembrane cells, and the expression of PIK3R1, ESR1, SRC, p-PI3K, and p-AKT were negatively correlated with the increase in Iso concentration (P < 0.05). Conclusion Iso can inhibit PI3K/AKT signal transduction and influence the expression of PIK3R1, AKT1, SRC, and ESR1 proteins, thereby inhibiting the occurrence and development of OSCC.

Objective To observe the application value of ultrasound classification of blood vessels at C6 transverse process region during ultrasound-guided cervical sympathetic block(CSB).Methods A total of 42 pain patients were retrospectively enrolled.According to vascular distribution morphology and variation,as well as the minimum vertical distance from carotid artery to C6 transverse process shown by ultrasound,blood vessels at C6 transverse process region was classified into type Ⅰ(standard type),Ⅱ(short arterial spacing type),Ⅲ(puncture path obstruction type)and Ⅳ(vascular variation type in blockade zone).CSB needle insertion strategy was adjusted according to the results of ultrasound classification.The needle tip directly reached the puncture target for type Ⅰ,"fluid push method"was used for type Ⅱ and type Ⅲ,while the needle tip needed to be guided to reach the area between carotid sheath and variant blood vessels in the puncture target area for type Ⅳ.The effectiveness and safety of this method were observed.Results Among 42 cases,type Ⅰ was observed in 18 cases(18/42,42.86%),type Ⅱ was noticed in 3 cases(3/42,7.14%),and type Ⅲ and Ⅳ was detected in 9(9/42,21.43%)and 12(12/42,28.57%)cases,respectively.CSB was successfully performed in all 42 cases,Horner syndrome was successfully induced and relieved spontaneously within 1-2 h.No adverse reactions such as local hematoma,pneumothorax,nor local pain was observed.Conclusion Ultrasound classification of blood vessels at C6 transverse process region during CSB was effective and safe,having certain promotion value.

Objective To investigate the mechanism of DNA methyltransferase 1(DNMT1)inhibitor combined with extracellular signal-regulated kinase 1(ERK1),homeodomain-interacting protein kinase 2(HIPK2),and glycogen synthase kinase 3 β(GSK3 β)inhibitors in synergistically inducing apoptosis and protein arrest in relapsed and refractory acute myeloid leukemia(AML)cells.Methods Three therapeutic targets,including ERK1,HIPK2,and GSK3β,were screened based on the Gene Expression Omnibus(GEO),The Cancer Genome Atlas(TCGA)database,and Ex-pression 2 Kinases database.Human AML cell line U937 cells were treated with DNMT1 inhibitor a-lone or combined with ERK1,HIPK2,or GSK3β inhibitors.Cell viability was detected using the CCK-8 method.Apoptosis rate was analyzed by flow cytometry,and cell cycle distribution was de-termined by propidium iodide(PI)staining.The mRNA expression levels of DNMT1,ERK1,HIPK2,and GSK3β were detected by real-time fluorescent quantitative reverse transcriptionpoly-merase chain reaction(RT-qPCR).Protein expressionlevels of DNMT1,ERK1,HIPK2,and GSK3β were detected by immunoblotting.Results DNMT1 inhibitor significantly inhibited the cell viability of U937 cells,and significantly induced apoptosis and cell cycle arrest in U937 cells(P＜0.05).When DNMT1 inhibitor was combined with ERK1,HIPK2,or GSK3β inhibitors,cell via-bility and apoptosis rate were significantly reduced(P＜0.05).DNMT1 inhibitor alone or its com-bination with ERK1,HIPK2,and GSK3β inhibitors induced U937 cell arrest in the G0/G,phase,with a significant increase in the proportion of cells in the G0/G1 phase in the combination group(P＜0.05).The combination of DNMT1 inhibitor with ERK1,HIPK2,and GSK3β inhibitors sig-nificantly reduced the mRNA expression levels in DNMT1,ERK1,HIPK2,and GSK3β in U937 cells(P＜0.05).Similarly,the combination therapy significantly reduced the protein expression levels of DNMT1,ERK1,HIPK2,and GSK3β in U937 cells(P＜0.05).Conclusion DNMT1 inhibitor combined with ERK1,HIPK2,and GSK3 β inhibitors can synergistically induce apoptosis and protein arrest in relapsed and refractory AML cells,providing a novel strategy for combined tar-geted therapy of AML.

This study was aimed at exploring the epidemic characteristics and spatio-temporal evolution patterns of severe fever with thrombocytopenia syndrome(SFTS)in Henan Province from 2011 to 2022,to provide a reference for prevention and control ef-forts.Data on SFTS in Henan Province from 2011 to 2022 were collected from the Information Management System for Infectious Dis-ease Reporting,and epidemiological characteristics and spatio-temporal evolution patterns were investigated through spatial autocorre-lation analysis,standard deviation ellipse analysis,and spatial-centered transfer curves,on a district-county basis.From 2011 to 2022,a total of 5 471 SFTS cases associated with 81 deaths were reported in Henan Province.The incidence rate showed an increasing trend(χ2trend=23.24,P<0.001),and the average annual incidence was 0.4 677/100 000.The case-fatality rate showed a decreasing trend from 2011 to 2019(χ2trend=8.86,P=0.003),but an increasing trend from 2020 to 2022(χ2trend=12.93,P<0.001).The average an-nual case-fatality rate was 1.48%.The peak incidence period was from May to July;this period accounted for 58.75%of the total cases.Females(59.39%),individuals 60-80 years old(55.40%),and farmers(96.64%)were the high prevalence groups.The disease was distributed primarily in Xinyang City,and Shangcheng County had the highest incidence rate(27.411 8/100 000).Spatial aggregation was observed in the southeastern region(Moran's I>0,P<0.001),and the disease centers were all located in Guangshan County,Xin-yang City,and showed southeast-northwest-southeast-northwest-southeast-northeast-southwest movement during the 12-year pe-riod.In conclusion,from 2011 to 2022,SFTS in Henan Province had a high incidence in May to July,and females,older people,and farmers were the high-risk groups.Spatial aggregation was observed in the southeastern region,along with spread to surrounding areas.Xinyang City is therefore a key area for the prevention and control of SFTS.

Objective:To evaluate the role of hippocampal activating transcription factor 5 (ATF5) in cognitive impairment induced by neuropathic pain and the relationship with mitochondrial unfolded protein response(mtUPR) in mice.Methods:This study was conducted in 2 parts. Experiment Ⅰ Twenty-four SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 2 groups ( n=12 each) using a random number table method: sham operation group (S1 group) and neuropathic pain group (NP group). Neuropathic pain was induced by chronic constriction injury to the sciatic nerve. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before developing the model and at 7, 14, 21 and 28 days after developing the model. Mouse cognitive function was assessed using the novel object recognition test from 30-31 days after developing the model. After the end of the novel object recognition test, mice were sacrificed and the hippocampal CA1 region was harvested for determination of the expression of ATF5 (by Western blot) and the expression of ATF5 in neurons, microglia and astrocytes (by immunofluorescence double staining). Experiment Ⅱ Thirty-six SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation group (S2 group), neuropathic pain + ATF5 up-regulation group (NA group), and neuropathic pain + empty virus group (NE group). On day 14 after developing the model, a virus that specifically up-regulated ATF5 expression in neurons and empty virus were injected into the hippocampal CA1 region. The MWT and TWL were measured at days 28 and 35 after developing the model. The novel object recognition test was performed on day 36 after developing the model to evaluate the cognitive function. After the end of the behavioral test, mice were sacrificed and the hippocampal CA1 region was harvested for detection of the expression of ATF5 and mtUPR marker proteins (Lon protease [LONP1] and heat shock protein 60 [HSP60]) by Western blot. Results:Experiment Ⅰ Compared with S1 group, no statistically significant change was found in the MWT and TWL before developing the model ( P>0.05), the MWT and TWL were significantly decreased on days 7, 14, 21 and 28 after developing the model, the discrimination index (DI) was decreased at day 31 after developing the model, the expression of ATF5 was down-regulated, the expression of ATF5 in neurons was down-regulated ( P<0.05), and no statistically significant change was found in the expression of ATF5 in mircrolia and astrocytes in NP group ( P>0.05). Experiment Ⅱ Compared with S2 group, the MWT and TWL were significantly decreased on days 28 and 35 after developing the model in NE group and NA group, DI was decreased, and the expression of ATF5, LONP1 and HSP60 was down-regulated in NE group ( P<0.05), and no significant change was found in NA group ( P>0.05). Compared with NE group, no significant change was found in the MWT and TWL in NA group ( P>0.05), DI was significantly increased, and the expression of ATF5, LONP1 and HSP60 was up-regulated in NA group ( P<0.05). Conclusions:Down-regulated ATF5 in the hippocampus is involved in the process of cognitive impairment caused by neuropathic pain, and the mechanism may be related to the inhibition of mtUPR.

Debates persist regarding the efficacy and safety of azvudine, particularly its real-world outcomes. This study involved patients aged ≥60 years who were admitted to 25 hospitals in mainland China with confirmed SARS-CoV-2 infection between December 1, 2022, and February 28, 2023. Efficacy outcomes were all-cause mortality during hospitalization, the proportion of patients discharged with recovery, time to nucleic acid-negative conversion (T _NANC), time to symptom improvement (T _SI), and time of hospital stay (T _HS). Safety was also assessed. Among the 5884 participants identified, 1999 received azvudine, and 1999 matched controls were included after exclusion and propensity score matching. Azvudine recipients exhibited lower all-cause mortality compared with controls in the overall population (13.3% vs. 17.1%, RR, 0.78; 95% CI, 0.67-0.90; P = 0.001) and in the severe subgroup (25.7% vs. 33.7%; RR, 0.76; 95% CI, 0.66-0.88; P < 0.001). A higher proportion of patients discharged with recovery, and a shorter T _NANC were associated with azvudine recipients, especially in the severe subgroup. The incidence of adverse events in azvudine recipients was comparable to that in the control group (2.3% vs. 1.7%, P = 0.170). In conclusion, azvudine showed efficacy and safety in older patients hospitalized with COVID-19 during the SARS-CoV-2 omicron wave in China.

Objective:To evaluate the role of hippocampal activating transcription factor 5 (ATF5) in cognitive impairment induced by neuropathic pain and the relationship with mitochondrial unfolded protein response(mtUPR) in mice.Methods:This study was conducted in 2 parts. Experiment Ⅰ Twenty-four SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 2 groups ( n=12 each) using a random number table method: sham operation group (S1 group) and neuropathic pain group (NP group). Neuropathic pain was induced by chronic constriction injury to the sciatic nerve. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before developing the model and at 7, 14, 21 and 28 days after developing the model. Mouse cognitive function was assessed using the novel object recognition test from 30-31 days after developing the model. After the end of the novel object recognition test, mice were sacrificed and the hippocampal CA1 region was harvested for determination of the expression of ATF5 (by Western blot) and the expression of ATF5 in neurons, microglia and astrocytes (by immunofluorescence double staining). Experiment Ⅱ Thirty-six SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation group (S2 group), neuropathic pain + ATF5 up-regulation group (NA group), and neuropathic pain + empty virus group (NE group). On day 14 after developing the model, a virus that specifically up-regulated ATF5 expression in neurons and empty virus were injected into the hippocampal CA1 region. The MWT and TWL were measured at days 28 and 35 after developing the model. The novel object recognition test was performed on day 36 after developing the model to evaluate the cognitive function. After the end of the behavioral test, mice were sacrificed and the hippocampal CA1 region was harvested for detection of the expression of ATF5 and mtUPR marker proteins (Lon protease [LONP1] and heat shock protein 60 [HSP60]) by Western blot. Results:Experiment Ⅰ Compared with S1 group, no statistically significant change was found in the MWT and TWL before developing the model ( P>0.05), the MWT and TWL were significantly decreased on days 7, 14, 21 and 28 after developing the model, the discrimination index (DI) was decreased at day 31 after developing the model, the expression of ATF5 was down-regulated, the expression of ATF5 in neurons was down-regulated ( P<0.05), and no statistically significant change was found in the expression of ATF5 in mircrolia and astrocytes in NP group ( P>0.05). Experiment Ⅱ Compared with S2 group, the MWT and TWL were significantly decreased on days 28 and 35 after developing the model in NE group and NA group, DI was decreased, and the expression of ATF5, LONP1 and HSP60 was down-regulated in NE group ( P<0.05), and no significant change was found in NA group ( P>0.05). Compared with NE group, no significant change was found in the MWT and TWL in NA group ( P>0.05), DI was significantly increased, and the expression of ATF5, LONP1 and HSP60 was up-regulated in NA group ( P<0.05). Conclusions:Down-regulated ATF5 in the hippocampus is involved in the process of cognitive impairment caused by neuropathic pain, and the mechanism may be related to the inhibition of mtUPR.

This study was aimed at exploring the epidemic characteristics and spatio-temporal evolution patterns of severe fever with thrombocytopenia syndrome(SFTS)in Henan Province from 2011 to 2022,to provide a reference for prevention and control ef-forts.Data on SFTS in Henan Province from 2011 to 2022 were collected from the Information Management System for Infectious Dis-ease Reporting,and epidemiological characteristics and spatio-temporal evolution patterns were investigated through spatial autocorre-lation analysis,standard deviation ellipse analysis,and spatial-centered transfer curves,on a district-county basis.From 2011 to 2022,a total of 5 471 SFTS cases associated with 81 deaths were reported in Henan Province.The incidence rate showed an increasing trend(χ2trend=23.24,P<0.001),and the average annual incidence was 0.4 677/100 000.The case-fatality rate showed a decreasing trend from 2011 to 2019(χ2trend=8.86,P=0.003),but an increasing trend from 2020 to 2022(χ2trend=12.93,P<0.001).The average an-nual case-fatality rate was 1.48%.The peak incidence period was from May to July;this period accounted for 58.75%of the total cases.Females(59.39%),individuals 60-80 years old(55.40%),and farmers(96.64%)were the high prevalence groups.The disease was distributed primarily in Xinyang City,and Shangcheng County had the highest incidence rate(27.411 8/100 000).Spatial aggregation was observed in the southeastern region(Moran's I>0,P<0.001),and the disease centers were all located in Guangshan County,Xin-yang City,and showed southeast-northwest-southeast-northwest-southeast-northeast-southwest movement during the 12-year pe-riod.In conclusion,from 2011 to 2022,SFTS in Henan Province had a high incidence in May to July,and females,older people,and farmers were the high-risk groups.Spatial aggregation was observed in the southeastern region,along with spread to surrounding areas.Xinyang City is therefore a key area for the prevention and control of SFTS.

Objective To observe the application value of ultrasound classification of blood vessels at C6 transverse process region during ultrasound-guided cervical sympathetic block(CSB).Methods A total of 42 pain patients were retrospectively enrolled.According to vascular distribution morphology and variation,as well as the minimum vertical distance from carotid artery to C6 transverse process shown by ultrasound,blood vessels at C6 transverse process region was classified into type Ⅰ(standard type),Ⅱ(short arterial spacing type),Ⅲ(puncture path obstruction type)and Ⅳ(vascular variation type in blockade zone).CSB needle insertion strategy was adjusted according to the results of ultrasound classification.The needle tip directly reached the puncture target for type Ⅰ,"fluid push method"was used for type Ⅱ and type Ⅲ,while the needle tip needed to be guided to reach the area between carotid sheath and variant blood vessels in the puncture target area for type Ⅳ.The effectiveness and safety of this method were observed.Results Among 42 cases,type Ⅰ was observed in 18 cases(18/42,42.86%),type Ⅱ was noticed in 3 cases(3/42,7.14%),and type Ⅲ and Ⅳ was detected in 9(9/42,21.43%)and 12(12/42,28.57%)cases,respectively.CSB was successfully performed in all 42 cases,Horner syndrome was successfully induced and relieved spontaneously within 1-2 h.No adverse reactions such as local hematoma,pneumothorax,nor local pain was observed.Conclusion Ultrasound classification of blood vessels at C6 transverse process region during CSB was effective and safe,having certain promotion value.

Tumor vaccine is a promising strategy for cancer immunotherapy by introducing tumor antigens into the body to activate specific anti-tumor immune responses. Along with the technological breakthroughs in genetic engineering and delivery systems, messenger ribonucleic acid (mRNA) technology has achieved unprecedented development and application over the last few years, especially the emergency use authorizations of two mRNA vaccines during the COVID-19 pandemic, which has saved countless lives and makes the world witness the powerful efficacy of mRNA technology in vaccines. However, unlike infectious disease vaccines, which mainly induce humoral immunity, tumor vaccines also need to activate potent cellular immunity to control tumor growth, which creates a higher demand for mRNA delivery to the lymphatic organs and antigen-presenting cells (APCs). Here we review the existing bottlenecks of mRNA tumor vaccines and advanced nano-based strategies to overcome those challenges, as well as future considerations of mRNA tumor vaccines and their delivery systems.