OBJECTIVE To study the effects of sophoranone （SOP） on the biological behavior of nasopharyngeal carcinoma CNE-1 cells and mitogen-activated protein kinase （MAPK） signaling pathway. METHODS CNE-1 cells were divided into blank group and SOP low-， medium- and high-concentration groups （SOP-L group， SOP-M group， SOP-H group， 25， 50 and 100 μmol/L）. The number of invasive cells， the number of migratory cells， and the apoptosis rate of cells were detected. The expression levels of mitogen-activated protein kinase kinase （MEK）， extracellular signal-regulated kinase 1 （ERK1）， ERK2， and c-Jun N-terminal kinase （JNK） mRNA， as well as phosphorylation levels of ERK， JNK， and p38 mitogen-activated protein kinase （abbreviated as “p38”） proteins in cells were all detected. Additionally， cells were divided into blank group， SOP high-concentration group （SOP- H group， 100 μmol/L）， SOP high-concentration combined with p38 inhibitor group （SOP-H+SB group， 100 μmol/L SOP+10 μmol/L SB）， and SOP high-concentration combined with JNK inhibitor group （SOP-H+SP group， 100 μmol/L SOP+10 μmol/L SP）. The number of invasive cells， cell migration rate， and the protein phosphorylation levels of JNK and p38 in cells， as well as the protein expression levels of matrix metalloproteinase-9（MMP-9）， proliferating cell nuclear antigen Ki67， and cleaved-caspase-3 were measured. RESULTS Compared with the blank group， SOP for each concentration could significantly decrease the number of invasive cells， the number of migratory cells， and mRNA expressions of MEK， ERK1， ERK2 （except for the SOP-L group） and JNK， but increase the apoptosis rate of cells and phosphorylation levels of ERK， JNK， and p38 proteins （P＜0.05）. Compared with the SOP-H group， the protein phosphorylation levels of p38 and JNK， and the protein expression of cleaved-caspase-3 were decreased significantly in SOP-H+SB group and SOP-H+SP group， while the number of invasive cells， cell migration rate， and the protein expression levels of MMP-9 and Ki67 were all increased significantly （P＜0.05）. CONCLUSIONS SOP can inhibit the proliferation， migration and invasion of CNE-1 cells， and induce the apoptosis， the mechanisms of which may be associated with promoting the phosphorylation of proteins related to the MAPK signaling pathway.

Aim To explore the efficacy of levosimendan on hypoxia pulmonary hypertension through animal experiments, and to further explore the potential mechanism of action using network pharmacological methods and molecular docking technique. Methods The rat model of hypoxia pulmonary hypertension was constructed to detect right heart systolic pressure and right heart remodeling index. HE , Masson, and VG staining were core targets were screened out. GO and KEGG pathway enrichment analysis were performed using the DAVID database. Molecular docking of the core targets was performed with the AutoDock software. Results The results of animal experiments showed that levosimendan had obvious therapeutic effect on hypoxia pulmonary hypertension. The network pharmacology results showed that SRC, HSP90AA1, MAPK1, PIK3R1, AKT1, HRAS, MAPK14, LCK, EGFR and ESR1 used to analyze the changes of rat lung histopathology. Search the Swiss Target Prediction, DrugBank Online, BatMan, Targetnet, SEA, and PharmMapper databases were used to screen for drug targets. Disease targets were retrieved from the GeneCards, OMIM databases. The "drug-target-disease" network was constructed after identification of the two intersection targets. The protein interaction network was constructed and the were the key targets to play a therapeutic role. Molecular docking showed good docking of levosimendan with all the top five core targets with degree values. Conclusions Levosimendan may exert a therapeutic effect on hypoxia-induced pulmonary hypertension through multiple targets.

Objective To investigate the effect of butorphanol on the malignant biological behavior of glioma cells by regulating the sonic hedgehog (SHH)/glioma-associated oncogene homolog 1 (GLI1) signaling pathway. Methods Glioma LN229 cells were divided into control group, low-dose butorphanol group, medium-dose butorphanol group, high-dose butorphanol group, high-dose butorphanol+pcDNA3.1 group and high-dose butorphanol+pc-SHH group. MTT and Edu assays were used to detect cell proliferation; flow cytometry was used to detect cell apoptosis rate; Transwell chamber assay was used to detect cell migration and invasion; real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect SHH mRNA and GLI1 mRNA expression levels; Western blot was used to detect the protein expression levels of Ki-67, matrix metalloproteinase-2 (MMP-2), cleaved caspase-3, SHH and GLI1. Results Butorphanol could reduce the viability and proliferation rate of LN229 cells, and decreased the number of cell migration and invasion (P < 0.05). Butorphanol could reduce the expression levels of SHH mRNA, GLI1 mRNA and the protein expression of Ki-67, MMP-2, SHH as well as GLI1 in LN229 cells, promote cell apoptosis and increase the protein expression of cleaved caspase-3 (P < 0.05). Overexpression of SHH could partially reverse the inhibitory effect of butorphanol on LN229 cells (P < 0.05). Conclusion Butorphanol can inhibit the malignant biological behavior of glioma cells, and its mechanism may be related to the inhibition of the SHH/GLI1 signaling pathway.

Viruses are powerful tools for the study of modern neurosciences. Most of the research on the connection and function of neurons were done by using recombinant viruses, among which neurotropic herpesvirus is one of the most important tools. With the continuous development of genetic engineering and molecular biology techniques, several recombinant neurophilic herpesviruses have been engineered into different viral tools for neuroscience research. This review describes and discusses several common and widely used neurophilic herpesviruses as nerve conduction tracers, viral vectors for neurological diseases, and lytic viruses for neuro-oncology applications, which provides a reference for further exploring the function of neurophilic herpesviruses.
Herpesviridae/genetics* ; Neurosciences ; Genetic Vectors/genetics* ; Genetic Engineering ; Neurons

ObjectiveTo analyze the induction effect of Fusobacterium nucleatum （Fn） on endoplasmic reticulum stress-related proteins Glucose-regulating protein 78（GRP78） and X-box binding protein 1（XBP1） in esophageal squamous cell carcinoma （ESCC）， and to explore its potential mechanism and clinical significance. MethodsESCC cells KYSE150 and KYSE140 were infected with Fn for 12 h， 24 h and 48 h. The oxidative stress indexes （ROS， MDA and SOD） and the expression of GRP78 and XBP1 in each group were detected by oxidative stress index kit and Western blot. The experiment was divided into Fn groups， Fn+siNC1 groups， Fn+siGRP78 groups， Fn+siNC2 groups and Fn+siXBP1 groups； the oxidative stress indexes， paclitaxel （PTX） response efficacy， abilities of proliferation， invasion and metastasis in each group were compared. The infection of Fn and the expression of GRP78 and XBP1 in 234 ESCC and paracancerous tissues were detected by RNA scope and immunohistochemistry. The correlation between each factor and clinicopathological characteristics of patients was analyzed by Chi-square test. The influence of each factor on the survival of patients was compared by Kaplan-meier survival estimate. ResultsCompared with Fn uninfected KYSE150 and KYSE140 cells， the content of ROS and MDA was gradually increased， the activity of SOD was gradually decreased， and the expression of GRP78 and XBP1 was gradually increased in Fn infected groups （12 h， 24 h and 48 h） （P < 0.05）. Compared with Fn groups， Fn+siNC1 groups， and Fn+siNC2 groups， ROS and MDA contents were decreased， SOD activity was increased， PTX response efficacy was enhanced， and abilities of proliferation， invasion and metastasis were decreased in Fn+siGRP78 and Fn+siXBP1 groups （P < 0.05）. The rates of Fn， GRP78 and XBP1 in ESCC tissues were 43.16%， 69.66% and 60.68%， respectively. And the three indexes were significantly consistent （P < 0.05）. The patients with positive Fn infection and high expression of GRP78 and XBP1 were mostly males with a history of smoking and drinking， and the tumor differentiation degree was low， the invasion degree was deep， the lymph node metastasis rate was high， and the clinical stage was mostly stage Ⅲ/Ⅳ. The 5-year survival time of patients with above positive indexes was shortened （P < 0.05）. ConclusionsFn could induce endoplasmic reticulum stress by inducing the high expression of GRP78 and XBP1， and promote the malignant evolution of ESCC.

Objective: To investigate the molecular mechanism of circZNF609 targeting miR-153 to regulate the proliferation and apoptosis of diffuse large B-cell lymphoma. Methods: Fifty cases of lymphoma tissue from patients with diffuse large B-cell lymphoma who were diagnosed and treated in the First Affiliated Hospital of Zhengzhou University from July 2018 to December 2019 were collected. Thirty cases of normal lymph node tissues that were confirmed to be reactive hyperplasia by pathological diagnosis during the same period were selected as controls. Real time quantitative polymerase chain reaction (PCR) was used to detect the expression of circZNF609 in diffuse large B-cell lymphoma tissues and control hyperplasia lymph nodes. Diffuse large B-cell lymphoma OCI-LY19 cells were divided into control group (blank control), si-con group (transfected with siRNA control), si-ZNF609 group (transfected with circZNF609 siRNA), and si-ZNF609+ Anti-NC group (co-transfected with circZNF609 siRNA and inhibitor control) and si-ZNF609+ Anti-miR-153 group (co-transfected with circZNF609 siRNA and miR-153 inhibitor). Cell counting kit-8 (CCK-8) was used to detected proliferation, flow cytometry was used to detect cell cycle and apoptosis. Western blot was used to detect the protein expressions of C-caspase-3, cyclin D1, p21. The luciferase reporter system was used to identifie the relationship between circZNF609 and miR-153. Results: The expression level of circZNF609 in diffuse large B-cell lymphoma tissue was (1.44±0.22), higher than (0.37±0.14) in the control tissues (P<0.001). The cell survival rate of the si-ZNF609 group was (51.74±6.39)%, lower than (100.00±10.23)% of the control group and the (99.64±11.67)% of the si-con group (P<0.001). The proportion of cells in the G(0)/G(1) phase was (63.25±4.11)%, higher than (48.62±4.32)% of the control group and (47.12±3.20)% of the si-con group (P<0.001), the apoptosis rate was (13.36±1.42)%, higher than (3.65±0.47)% of the control group and (3.84±0.62)% of the si-con group (P<0.05). The expression levels of C-caspase-3 and p21 protein were (0.85±0.09) and (0.90±0.08), higher than (0.38±0.04) and (0.65±0.07) in the control group and (0.39±0.05) and (0.66±0.05) in the si-con group (P<0.001). The expression level of cyclin D1 protein was (0.40±0.03), lower than (0.52±0.06) of the control group and (0.53±0.04) of the si-con group (all P<0.001). CircZNF609 and miR-153 are mutually targeted. The cell survival rate of the si-ZNF609+ Anti-miR-153 group was (169.92±13.25)%, higher than (100.00±9.68)% of the si-ZNF609+ Anti-NC group (P<0.001), the ratio of cells in G(0)/G(1) phase and apoptosis rate were (52.01±3.62)% and (8.20±0.87)%, respectively, lower than (64.51±5.17)% and (14.03±1.17)% in the si-ZNF609+ Anti-NC group (P<0.001). The protein expression levels of C-caspase-3 and p21 were (0.42±0.06) and (0.52±0.06), lower than (0.80±0.07) and (0.92±0.10) of the si-ZNF609+ Anti-NC group (P<0.001). The protein expression level of cyclin D1 was (0.68±0.07), higher than (0.39±0.04) in the si-ZNF609+ Anti-NC group (P<0.001). Conclusion: Down-regulation of circZNF609 inhibits the proliferation of diffuse large B-cell lymphoma OCI-LY19 cells and induces apoptosis by targeting miR-153.
Apoptosis/genetics* ; Cell Line, Tumor ; Cell Proliferation/genetics* ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphoma, Large B-Cell, Diffuse/pathology* ; MicroRNAs/genetics* ; RNA, Circular/genetics*

ObjectiveTo evaluate the clinical efficacy and safety of Yangxin Dawayimicol honey ointment （YDHO） in the treatment of insomnia with the syndrome of Qi stagnation and blood stasis. MethodEighty insomnia patients who met the inclusion criteria in the Department of Encephalopathy of the Third Affiliated Hospital of Henan University of Chinese Medicine from November 2019 to October 2020 were randomly divided into an experimental group （48 cases） and a control group （32 cases）. The experimental group was treated with YDHO + Xuefu Zhuyu capsule simulators，and the control group was treated with Xuefu Zhuyu capsules + YDHO simulators for eight weeks. The changes in Pittsburgh sleep quality index（PSQI）score，traditional Chinese medicine （TCM） syndrome score，insomnia severity index （ISI），neurotransmitter indexes ［γ-aminobutyric acid（GABA），glutamic acid（Glu），and 5-hydroxy tryptamine（5-HT）］，serum inflammatory indexes ［interleukin-6（IL-6）and interleukin-10（IL-10）］， and safety index of the two groups were compared. ResultThe total effective rate was 97.83%（45/46） in the experimental group， higher than 68.75%（22/32） in the control group（Z=-4.292，P<0.01）. The experimental group was superior to the control group in PSQI score，ISI score，TCM syndrome score， and sleep duration（P<0.05）. The curative effects were equivalent between the two groups in shortening the time to fall asleep. The experimental group showed increased serum content of GABA，5-HT， and IL-10 and reduced content of Glu and IL-6，with few adverse reactions （P<0.05）. ConclusionYDHO is effective，safe， and reliable in the treatment of insomnia with Qi stagnation and blood stasis syndrome.

Objective: To explore the effects and mechanisms of interfering with transient receptor potential melastatin subfamily member 7(TRPM7) on biological behaviors(proliferation, apoptosis, invasion) of laryngeal carcinoma cells. Methods: Human laryngeal carcinoma cells TU212 were culturedin vitro. TRPM7-shRNA plasmid vectors(TRPM7-shRNA1 group, TRPM7-shRNA2 group, TRPM7-shRNA3 group) and negative control shRNA-NC(shRNA-NC group) were constructed. TU212 cells were transfected by liposome transfection method. The cells transfected with empty vector were enrolled as Control group. The level of lactic dehydrogenase(LDH) in TU212 supernatant was detected by ELISA. The level of malondialdehyde(MDA) and activity of superoxide dismutase(SOD) in supernatant were detected by colorimetry. The effects of knocking-down TRPM7 on proliferation and invasion of TU212 cells were detected by CCK-8, clone formation assay and Transwell assay. The expressions of invasion and apoptosis-related proteins were detected by Western blot. Results: After transfection, expression levels of TRPM7 mRNA and protein were down-regulated in TRPM7-shRNA1 group, TRPM7-shRNA2 group and TRPM7-shRNA3 group compared with Control group(P<0.05), and the decrease was the most significant in TRPM7-shRNA1 group(P<0.05). In functional experiments, SOD level in TRPM7-shRNA1 group decreased compared with Control group(P<0.05), while MDA and LDH levels increased(P<0.05). The cells proliferation rate and clone formation rate were decreased(P<0.05), the number of invasion cells was reduced(P<0.05), the expressions of N-cadherin and Vimentin proteins were down-regulated(P<0.05), mitochondrial membrane potentials were reduced(P<0.05), Bax/Bcl-2 and cleaved caspase-3/caspase-3 increased(P<0.05). Conclusion Knocking-down TRPM7 can increase oxidative stress level in laryngeal carcinoma cells TU212, inhibit their proliferation and invasion, and induce their apoptosis by mitochondrial pathways.

Angelicae Sinensis Radix， derived from a medicinal and edible plant Angelica sinensis， is one of the traditional bulk Chinese medicines. In addition to gynecological blood stagnation and deficiency， its indications also include dysmenorrhea， deficiency and cold-induced abdominal pain， and rheumatoid arthritis. With the in-depth study of Angelicae Sinensis Radix， its anti-inflammatory and analgesic activities have attracted widespread attention. However， there has been no systemic report. The present study comprehensively reviewed the anti-inflammatory and analgesic activities of Angelicae Sinensis Radix （including its compositions， extracts， and different processed products） and mechanisms published in recent 30 years. The anti-inflammatory effect of Angelicae Sinensis Radix was achieved mainly by blocking the expression of proteins and genes in nuclear factor-κB （NF-κB）， mitogen-activated protein kinases （MAPK）， and Janus kinase （JAK）/signal transducer and activator of transcription （STAT） signaling pathways， inhibiting the release of inflammatory mediators including tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， nitric oxide （NO）， prostaglandin E₂ （PGE₂） and IL-1β， and maintaining the high sensitivity of immune cells in the host to external stimuli. The mechanism of analgesic effect may be related to the suppression of the production of algesic substances （such as inflammatory factors and chemokines） or blocking of the amplification and transmission of pain perception in cascade reaction. Furthermore， the study also pointed out some problems in modern research and proposed suggestions on its future research to provide references for investigation and clinical applications of Angelicae Sinensis Radix.

【Objective】 To investigate the effects and potential mechanisms of neurodegenerative lesions in male mice caused by ozone exposure. 【Methods】 We divided 23 C57BL/6N male mice aged 8 to 9 months into control group （clean air group， 11） and ozone group （1 mg/m ³， 4h/d， 12）. After 8 weeks of continuous ozone exposure， the Morris water maze experiment was used to detect the mice’s learning and memory ability， HE dyeing to observe pathological changes in hippocampal tissue cells， and immunoprinting tests to detect the expression levels of Tau， p-Tau and α-synuclein proteins in the cerebral cortex tissue. 【Results】 After 8 weeks of ozone exposure， the mice’s spatial learning and memory ability were impaired to a certain extent， the incubation period decreased with time， and the two lines were separated， but the difference was not statistically significant. Ozone exposure caused changes in the morphology of the mice’s hippocampal tissue cells， disorders in the arrangement of hippocampal neuron， and nuclear wrinkles， and significantly increased levels of p-Tau and α-synuclein protein expressions in cerebral cortex tissues （P<0.01）， but there was no statistical significance in the total Tau expression level. 【Conclusion】 Ozone exposure leads to the loss of learning and memory in mice， changes in hippocampal neurocellular pathology， and increased expression levels of neurodegenerative variable-related proteins.