This study investigated the role of long non-coding RNAs (lncRNAs) in the development of the palatal tissues. Cleft palates in mice were induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Expression levels of long non-coding RNA H19 (lncRNA H19) and insulin-like growth factor 2 (IGF2) gene were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The rate of occurrence of cleft palate was found to be 100% by TCDD exposure, and TCDD could cause short upper limb, cerebral fissure, webbed neck, and short neck. The expression levels of lncRNA H19 and IGF2 gene specifically showed embryo age-related differences on E13, E14, and E15 in the palatal tissues. The expression levels of lncRNA H19 and IGF2 gene showed an inverse relationship on E13, E14, and E15. These findings demonstrated that lncRNA H19 and IGF2 can mediate the development of mouse cleft palate.
Animals ; Cleft Palate ; chemically induced ; genetics ; pathology ; Female ; Gene Expression Regulation ; Gene Expression Regulation, Developmental ; drug effects ; Male ; Mice ; Mice, Inbred C57BL ; Palate ; metabolism ; Polychlorinated Dibenzodioxins ; toxicity ; RNA, Long Noncoding ; genetics ; Real-Time Polymerase Chain Reaction

OBJECTIVE: To investigate the effect and mechanism of glucosides of chaenomeles speciosa (GCS) on ischemia/reperfusion-induced brain injury in mouse model. METHODS: Fifty 8-week C57BL/C mice were randomly divided into five groups with 10 in each group:sham group, model group, GCS 30 mg/kg group, GCS 60 mg/kg group and GCS 90 mg/kg group, and the GCS was administrated by gavage (once a day) for 14 d. HE staining was performed to investigate the cell morphology; the Zea-Longa scores were measured for neurological activity; TUNEL staining was performed to investigate the cell apoptosis; ELISA was used to detected the oxidative stress and inflammation; Western Blot was performed to investigate the key pathway and neurological functional molecules. RESULTS: Compared with the sham group, the brain tissues in model group were seriously damaged, presenting severe cell apoptosis, oxidative stress and inflammation, associated with increased NF-κB P65 and TNF-α levels as well as decreased myelin associate glycoprotein (MAG) and oligodendrocyte-myelin glycoprotein (OMgp)levels (all <0.01). Compared with the model group, the brain tissues in GCS groups were ameliorated, and cell apoptosis, oxidative stress and inflammation were inhibited, associated with decreased NF-κB P65 and TNF-α levels as well as increased MAG and OMgp levels (all <0.01), which were more markedly in GCS 60 mg/kg group. CONCLUSIONS GCS can inhibit the NF-κB P65 and TNF-α, reduce the oxidative stress and inflammation, decrease the cell apoptosis in mouse ischemia/reperfusion-induced brain injury model, and 60 mg/kg GCS may be the optimal dose.
Animals ; Brain ; drug effects ; Brain Injuries ; drug therapy ; Gene Expression Regulation ; drug effects ; Glucosides ; pharmacology ; therapeutic use ; Mice ; Mice, Inbred C57BL ; NF-kappa B ; genetics ; Oxidative Stress ; drug effects ; Plant Extracts ; pharmacology ; Random Allocation ; Rosaceae ; chemistry ; Tumor Necrosis Factor-alpha ; genetics

Angelicae Sinensis Radix, as a medicinal and edible Chinese medicinal herb, is widely used in clinical practice. It is mainly cultivated in Minxian, Tanchang, Zhangxian and Weiyuan counties of Gansu province. In recent years, with the comprehensive and in-depth study of Angelicae Sinensis Radix in China and abroad, its chemical composition, pharmacological effects and application and development have attracted much attention. In this study, the chemical composition, traditional efficacy, and modern pharmacological effects of Angelicae Sinensis Radix were summarized. On this basis, combined with the core concept of quality markers(Q-markers), the Q-markers of Angelicae Sinensis Radix were discussed from the aspects of mass transfer and traceability and chemical composition specificity, availability, and measurability, which provided scientific basis for the quality evaluation of Angelicae Sinensis Radix.
Angelica sinensis/chemistry* ; Drugs, Chinese Herbal/pharmacology* ; Plant Roots/chemistry* ; China

BACKGROUND: Neuroimaging studies have shown that chronic pain could cause functional and structural alterations; however, the neural mechanism of psychosomatic symptoms caused by chronic discogenic low back pain (CDLBP) remains unclear. OBJECTIVE: To investigate the alterations of amplitude of low frequency fluctuation (ALFF) of resting spontaneous brain activity in CDLBP patients and their relationships with clinical symptoms. METHODS: Resting-state brain functional MRI scans were performed in 67 CDLBP patients (CDLBP group) and 78 age-, sex-and education level-matched healthy controls (control group). The disease course, Visual Analogue Scale score, Oswestry Disability Index and Self-rating Depression Scale score were analyzed for CDLBP patients. The average ALFF values of the two groups were calculated using DPARSF and REST software, the differences of ALFF values between the two groups were compared by independent two-sample t-test, and the relationships between the ALFF values derived from significant regions and the course of disease and clinical symptoms were analyzed. The study protocol was approved by the Ethics Committee of Second Affiliated Hospital of Xinxiang Medical University (approval No. 20150210). RESULTS AND CONCLUSION: Compared with the control group, the regions with significantly increased ALFF values in the CDLBP group included the left insular lobe, right parahippocampal gyrus, bilateral superior temporal gyrus, left inferior temporal gyrus, bilateral prefrontal lobe, left caudate nucleus and right inferior frontal gyrus (all P < 0.05, Gaussian random field (GRF) corrected). Compared with the control group, the regions with significantly decreased ALFF values in the CDLBP group included the right superior orbital frontal gyrus and right lingual gyrus (both P <0.05, GRF corrected). Correlation analysis showed that the mean ALFF values in the right inferior frontal gyrus and left caudate nucleus were positively correlated with the Visual Analogue Scale (all P < 0.05). All these findings suggest that patients with CDLBP have abnormal spontaneous brain activities in multiple brain regions, and these alterations are correlated with the degree of pain.

Objective: To investigate the levels of Th9 cells and interleukin-9 (IL-9), and to assess the regulatory activity of Th9/IL-9 to anti-tumor immune response in patients with gastric cancer. Methods: Thirty-four patients with gastric cancer who received operation in the First Affiliated Hospital of Xinxiang Medical University between October 2018 and August 2019 were included. Twenty individuals who received physical examination in the same period were also enrolled. Peripheral blood was collected, and then plasma and peripheral blood mononuclear cells (PBMCs) were isolated. Tumor-infiltrating lymphocytes (TILs) and autologous gastric cancer cells were isolated from resected gastric cancer tissues. CD4(+) T cells, CD8(+) T cells, and CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells were purified from PBMCs and TILs. Plasma IL-9 level was measured by enzyme linked immunosorbent assay (ELISA). The percentage of CD3(+) CD4(+) IL-9(+) Th9 cells in PBMCs and TILSs was assessed by flow cytometry. The mRNA levels of IL-9 and transcriptional factors purine-rich nucleic acid binding protein 1 (PU.1) were semi-quantified by real-time quantitative polymerase chain reaction (RT-qPCR). PBMCs and TILs from gastric cancer patients were stimulated with recombinant human IL-9. Cellular proliferation was measured by cell counting kit-8. The phosphorylation levels of signal transducer and activator of transcription 3 (STAT3) and STAT6 were investigated by western blot. Cytokine production was measured by ELISA. Purified CD8(+) T cells from TILs of gastric cancer patients were stimulated with recombinant human IL-9. CD8(+) T cells and autologous gastric cancer cells were cocultured in direct contact and indirect contact manner. The percentage of target cell death was calculated by measuring the lactate dehydrogenase (LDH) level. These cretion of γ-Interferon (γ-IFN) and tumor necrosis factor-α (TNF-α) was measured by ELISA. CD4(+) CCR4(-)CCR6(-)CXCR3(-)cells, CD8(+) T cells, and autologous gastric cancer cells were directly cocultured, and anti-IL-9 neutralizing antibody was added. The target cell death was measured. Results: The percentages of CD3(+) CD4(+) IL-9(+) Th9 cells in PBMCs of control group and PBMCs of gastric cancer group were (1.21±0.25)% and (1.14±0.19)%, respectively. The difference was not statistically significant (P=0.280). The percentage of CD3(+) CD4(+) IL-9(+) Th9 cells in TILs of gastric cancer group was (2.30±0.55)%, which was higher than those in PBMCs of control group and PBMCs of gastric cancer group (P<0.001). The plasma IL-9 level in control group and gastric cancer group were (5.04±1.51) and (4.93±1.25) ng/ml. The difference was not statistically significant (P=0.787). The relative levels of IL-9 mRNA in PBMCs of control group and PBMCs of gastric cancer group were 1.33±0.39 and 1.36±0.27. The difference was not statistically significant (P=0.691). The relative level of IL-9 mRNA in TILs of gastric cancer group was 2.90±0.75, which was higher than those in PBMCs of control group (P<0.001) and PBMCs of gastric cancer group (P<0.001). The relative levels of PU.1 mRNA in PBMCs of control group and PBMCs of gastric cancer group were 1.21±0.12 and 1.20±0.11. The difference was not statistically significant (t=0.21, P=0.833). PU.1 mRNA relative level in TILs of gastric cancer group was 2.81±0.65, which was higher than those in PBMCs of control group (P<0.001) and PBMCs of gastric cancer group (P<0.001). Recombinant human IL-9 stimulation did not affect the proliferation of PBMCs and TILs of gastric cancer patients (P>0.05), but elevated the phosphorylation level of STAT6 and induced the secretions of γ-IFN, IL-17, and IL-22 by TILs (P<0.05). In direct contact culture system, IL-9 stimulation promoted tumor-infiltrating CD8(+) T cells-induced autologous gastric cancer cell death [(20.62±2.27)% vs. (16.08±2.61)%, P<0.01)]. In indirect contact culture system, IL-9 stimulation did not increase CD8(+) T cell-induced autologous gastric cancer cell death [(5.21±0.70)% vs. (5.31±1.22)%, P=0.998)]. However, the secretion levels of γ-IFN were elevated in response to IL-9 stimulation in both culture systems [direct contact culture system: (100.40±12.05) pg/ml vs. (76.45±8.56) pg/ml; indirect contact culture system: (78.00±9.98) pg/ml vs. (42.09±10.71) pg/ml; P<0.01]. The TNF-α secretion level did not significantly changed (P>0.05). In direct contact culture system, the percentage of target cells was (22.01±3.05) % and γ-IFN secretion level was (104.5±12.84) pg/ml in CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells+ CD8(+) T cells+ gastric cancer cells group, which was higher than (16.08±2.61)% and (76.45±8.56) pg/ml in CD8(+) T cells+ gastric cancer cells group (P<0.01). However, the percentage of target cells was (14.47±3.14)% and γ-IFN secretion level was (70.45±19.43) pg/ml in CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells+ CD8(+) T cells+ gastric cancer cells+ anti-IL-9 neutralizing antibody group, which were lower than those in CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells+ CD8(+) T cells+ gastric cancer cells group (P<0.01). Conclusion: Tumor-infiltrating Th9 cells and the secreting IL-9 promote the activity of CD8(+) T cells in gastric cancer patients, and enhance anti-tumor immune response.
Humans ; CD8-Positive T-Lymphocytes ; Stomach Neoplasms/pathology* ; Tumor Necrosis Factor-alpha/metabolism* ; Lymphocytes, Tumor-Infiltrating/pathology* ; Interferon-gamma/metabolism* ; RNA, Messenger/metabolism* ; Antibodies, Neutralizing/metabolism*

Purpose: This study aimed to evaluate nursing students’ understanding of the prevention of COVID-19, as well as their anxiety towards the disease and their perception of their professional identity in the wake of the pandemic, in Zhengzhou, China. Methods: A cross-sectional study was designed to investigate 474 nursing students by cluster sampling using a stratified questionnaire from February 15 to March 31, 2020. Multiple linear regression was used to identify the factors affecting professional identity. Binary and multiple logistic regression were used to identify the factors affecting anxiety. Results: Responders with a high level of understanding of COVID-19 and frequent use of behavioral strategies for its prevention comprised 93.2% and 30.0% of the cohort, respectively. Professional identity was significantly associated with gender and anxiety (p < .050). The prevalence of anxiety among nursing students was 12.4%. Male (odds ratio [OR] = 2.39; 95% confidence interval [CI] = 1.26~4.52), sophomores (OR = 5.30; 95% CI = 1.61~7.45), and infrequent use of prevention measures (OR = 3.49; 95% CI = 1.16~5.19) had a significant effect on anxiety. Conclusion Anxiety during the COVID-19 epidemic gives an adverse effect on the professional identity of nursing in students. Nursing education institutions need to provide psychological counseling services for nursing students, in addition to improving their teaching of COVID-19 prevention strategies.

To study the role of oleanolic acid on interleukin (IL)-1β-stimulated expression of inflammatory cytokines, and to explore its anti-inflammatory mechanism in SW982 cells, the toxicity of oleanolic acid on SW982 cells was detected by MTT; effects of different concentrations of oleanolic acid (5, 10, 20 μmol·L^-1) on the expression of inflammatory factors IL-6, IL-8 and matrix metalloproteinase-1 (MMP-1) was tested at protein and mRNA levels. The study was performed in IL-1β-stimulated SW982 cells together with enzyme-linked immunosorbent assay (ELISA) and real-time fluorescence quantitative PCR (real-time PCR) methods; the influence of oleanolic acid on the phosphorylation of mitogen-activated protein kinase (MAPK), phosphatidyl inositol-3-kinase/Akt (PI3K/Akt) and nuclear transcription factor-κB (NF-κB) signaling pathways related protein was analyzed by Western blot. Results showed that different concentrations of oleanolic acid (≤ 40 μmol·L^-1) were almost non-toxicity to SW982 cells; oleanolic acid significantly inhibited the expression of inflammatory factors in a dose-dependent manner; oleanolic acid restrained extracellular signal-related kinase (ERK), p38, c-jun N-terminal kinase (JNK) and Akt protein phosphorylation and IκB-α protein degradation obviously. The inhibition effect of oleanolic acid on inflammatory factors stimulated by IL-1β may be worked through MAPK, PI3K/Akt and NF-κB signaling pathways.

【Objective】 To investigate the current situation and related factors of emotional and behavioral problems in children with diurnal urinary incontinence (DUI), in order to provide reference for clinical treatment and psychological intervention. 【Methods】 During Sep.2019 and Mar.2020, a survey was conducted in six primary schools in a county of Henan Province using the method of cluster sampling. The survey included general data and current DUI and urination-related problems, Strengths and Difficulties Scale (SDQ), and Pittsburgh Sleep Quality Index (PSQI). 【Results】 Of the 4 500 questionnaires distributed, 4 120 were collected, and 3 912 were qualified for statistical analysis. The overall prevalence of DUI was 3.6%. The detection rate of abnormal emotion and behavior in the DUI group was 48.2%, which was significantly higher than that in the non-DUI group (12.6%) (P<0.05). The scores of emotional symptoms, conduct problems, hyperactivity, peer communication problems and SDQ were significantly higher in the DUI group than in the control group (P<0.05). The results of multivariate analysis showed that gender, academic achievement, DUI times, caregivers, caregivers’ education level, family education style and sleep disorder were influencing factors of emotional and behavioral problems in DUI children (P<0.05). The total score of SDQ was positively correlated with the total score of PSQI (P<0.05). 【Conclusion】 The detection rate of emotional and behavioral abnormalities is high in DUI children, which is related to gender, academic achievement, DUI times, caregivers, education level of caregivers, family education style and sleep disorders.

Objective: To test the effect of metastasis associated in lung adenocarcinoma transcript 1 (MALAT1) and/or osimertinib on the proliferation and apoptosis of HCC827 cells, and explore the potential mechanism of MALAT1 induced resistance to osimertinib. Methods: We transfected HCC827 cells with LV-vector or LV-over/MALAT1. Stable transfected cells (HCC827/Vector, HCC827/MALAT1) were selected by adding puromycin. HCC827/MALAT1 cells were further transfected with the shRNA-negative control (NC) or shRNA-human epidermal growth factor receptor 3 (ERBB3) plasmid. The effects of overexpression of MALAT1, knockdown of ERBB3 and/or osimertinib on the proliferation of HCC827 cells were evaluated by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay. Cell apoptosis induced by MALAT1 overexpression, knockdown of ERBB3 and/or osimertinib treatment were analyzed by flow cytometry analysis. The expressions of EGFR and ERBB3 signal pathway related proteins in HCC827 cells treated with overexpression of MALAT1, knockdown of ERBB3 and/or osimertinib treatment were detected by western blot. Results: The MTT assay showed that sensitivity to osimertinib of HCC827/MALAT1 cells were significantly repressed. The 50% inhibitive concentration (IC₅₀) of osimertinib >4 000 nmol/L in HCC827/MALAT1 cells. However, knockdown of ERBB3 facilitated the anti-proliferation effect of osimertinib, and the IC₅₀ of osimertinib in shRNA-ERBB3 cells was (17.27±3.21) nmol/L. The results of flow cytometry analysis showed that the apoptotic rate of HCC827/MALAT1 cells induced by 10 nmol/L osimertinib was (8.38±0.92)%, significantly lower than (27.17±5.83)% of knockdown of ERBB3 (P<0.01). Western blotting showed that the expression of p-ERBB3, p-AKT and p-extracellular regulated protein kinases (ERK) in HCC827/MALAT1 cells was markedly up-regulated, while the expression of p-epithelial growth factor receptor (EGFR) was inhibited. The expressions of p-ERBB3, p-AKT and p-ERK were marginally affected by osimertinb. However, osimertinib downregulated the expressions of p-EGFR, p-ERBB3, p-AKT and p-ERK in ERBB3 deleted cells. Conclusions MALAT1 confers resistance to osimertinb in HCC827 cells by activating of the ERBB3/PI3K/AKT and ERBB3/MAPK/ERK signaling pathways.

This study was aimed to establish the cell lines of human T-cell leukemia virus type-1 (HTLV-1) tax gene expression and their biological activity. The eukaryotic vector pCMV-tax and pCMV-neo-Bam were transfected into Jurkat E6-1 by using liposome, following screening with G418, the tax gene expression cell line-TaxP and the negative cell lines-TaxN were selected and enriched. Then, the mRNA expressions of LAT, SLP70, ZAP70 and NF-kappaB (p65) were measured by using RT-PCR; the NF-kappaB bioactivity was tested after transfecting the pNF-kappaB-Luc plasmids into TaxP and TaxN cells; meanwhile CD25, CD69 expressions on surface of the two cell lines were tested by flow cytometry. The results showed that TaxP and TaxN cell lines were established and the tax gene expression was detected; as compared with TaxN, the mRNA expressions of LAT, SLP70 and ZAP70 in TaxP were enhanced (p<0.05), while the NF-kappaB bioactivity in TaxP cells was stronger than that in TaxN cells (p<0.01). The CD25, CD69 on TaxP cells surface was highly expressed. It is concluded that TaxP and TaxN cell lines are established, TAX protein could promote the activation of T cells and activate the NF-kappaB pathway.
Cell Line ; Gene Expression ; Gene Expression Regulation, Viral ; Genes, pX ; Human T-lymphotropic virus 1 ; genetics ; Humans ; Jurkat Cells ; NF-kappa B ; metabolism ; Transfection