Objective To explore the effects of deletion of protein 4.1R on hepatocyte proliferation,apoptosis,and glycolysis and the molecular mechanisms.Methods A 4.1R-/-HL-7702 cell line was constructed using CRISPR/Cas9 technique,and with 4.1R+/+HL-7702 cells as the control,its proliferative capacity and cell apoptosis were assessed using CCK-8 assay,EdU-488 staining,flow cytometry and Annexin V-FITC/PI staining at 24,48,72 h of cell culture.The changes in glucose uptake,lactate secretion,ATP production and pH value of the culture supernatant of 4.1R-/-HL-7702 cells were determined.The mRNA expressions of the key regulatory enzymes HK2,PFKL,PKM2 and LDHA in glycolysis were detected with qRT-PCR,and the protein expressions of AMPK,p-AMPK,Raptor and p-Raptor were determined using Western blotting.Results Western blotting and sequencing analysis both confirmed the successful construction of 4.1R-/-HL-7702 cell line.Compared with the wild-type cells,4.1R-/-HL-7702 cells exhibited a lowered proliferative activity with increased cell apoptosis.The deletion of protein 4.1R also resulted in significantly decreased glucose uptake,lactate secretion and ATP production of the cells and increased pH value of the cell culture supernatant.qRT-PCR showed significantly decreased mRNA expressions of the key regulatory enzymes in glycolysis in 4.1R-/-HL-7702 cells.Compared with those in HL-7702 cells,the expression levels of AMPK and Raptor proteins were decreased while the expression levels of p-AMPK and p-Raptor proteins increased significantly in 4.1R-/-HL-7702 cells.Conclusion Deletion of protein 4.1R in HL-7702 cells results in reduced proliferative capacity,increased apoptosis and suppression of glycolysis,and this regulatory mechanism is closely related with the activation of the downstream AMPK-mTORC1 signaling pathway.

Objective To explore the effects of deletion of protein 4.1R on hepatocyte proliferation,apoptosis,and glycolysis and the molecular mechanisms.Methods A 4.1R-/-HL-7702 cell line was constructed using CRISPR/Cas9 technique,and with 4.1R+/+HL-7702 cells as the control,its proliferative capacity and cell apoptosis were assessed using CCK-8 assay,EdU-488 staining,flow cytometry and Annexin V-FITC/PI staining at 24,48,72 h of cell culture.The changes in glucose uptake,lactate secretion,ATP production and pH value of the culture supernatant of 4.1R-/-HL-7702 cells were determined.The mRNA expressions of the key regulatory enzymes HK2,PFKL,PKM2 and LDHA in glycolysis were detected with qRT-PCR,and the protein expressions of AMPK,p-AMPK,Raptor and p-Raptor were determined using Western blotting.Results Western blotting and sequencing analysis both confirmed the successful construction of 4.1R-/-HL-7702 cell line.Compared with the wild-type cells,4.1R-/-HL-7702 cells exhibited a lowered proliferative activity with increased cell apoptosis.The deletion of protein 4.1R also resulted in significantly decreased glucose uptake,lactate secretion and ATP production of the cells and increased pH value of the cell culture supernatant.qRT-PCR showed significantly decreased mRNA expressions of the key regulatory enzymes in glycolysis in 4.1R-/-HL-7702 cells.Compared with those in HL-7702 cells,the expression levels of AMPK and Raptor proteins were decreased while the expression levels of p-AMPK and p-Raptor proteins increased significantly in 4.1R-/-HL-7702 cells.Conclusion Deletion of protein 4.1R in HL-7702 cells results in reduced proliferative capacity,increased apoptosis and suppression of glycolysis,and this regulatory mechanism is closely related with the activation of the downstream AMPK-mTORC1 signaling pathway.

Objective:This study aimed to investigate the change of the position of the tongue before and after combined treatment of maxillary expansion and orofacial myofunctional therapy in children with mouth-breathing and skeletal class Ⅱmalocclusion. Methods:A total of 30 children with skeletal class Ⅱ malocclusion and unobstructed upper airway were selected. The 30 children were divided into mouth-breathing group（n=15） and nasal-breathing group（n=15） and CBCT was taken. The images were measured by Invivo5 software. The measurement results of the tongue position of the two groups were analyzed by independent samples t-test. 15 mouth-breathing children with skeletal class Ⅱ malocclusion were selected for maxillary expansion and orofacial myofunctional therapy. CBCT was taken before and after treatment, the measurements were analyzed by paired sample t test with SPSS 27.0 software package. Results:The measurement of the tongue position of the mouth-breathing and nasal-breathing groups were compared, the differences were statistically significant（P<0.05）. The measurement of the tongue position showed significant difference after the combined treatment of maxillary expansion and orofacial myofunctional therapy in children with mouth-breathing and skeletal class Ⅱmalocclusion（P<0.05）. Conclusion:Skeletal class Ⅱ malocclusion children with mouth-breathing have low tongue posture. The combined treatment of maxillary expansion and orofacial myofunctional therapy can change the position of the tongue.
Child ; Humans ; Myofunctional Therapy/methods* ; Mouth Breathing/therapy* ; Palatal Expansion Technique ; Tongue ; Malocclusion/therapy*

Objective:To investigate the effects and mechanism of cannabinoid (CBD) on the anxiety- and depression-like behaviors induced by lipopolysaccharide (LPS) in mice.Methods:C57BL/6J mice were intraperitoneally injected with LPS to establish the model of neuroinflammation. CBD was injected intraperitoneally 24 h after modeling. Behavioral tests were performed to evaluate the anxiety- and depression-like behaviors in mice. CBD-pretreated BV-2 microglia cells were stimulated with LPS in vitro. The levels of tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β) and CD86 in mouse cerebral cortex, hippocampus, prefrontal cortex and BV-2 cells were measured by qRT-PCR. The protein level of nuclear factor (NF-κB) in mouse brain and BV-2 cells was determined by Western blot. Results:CBD significantly increased the residence time and movement distance of LPS-treated mice in the central area in the open filed test (OFT), and reduced the immobility time in tail suspension test (TST) and force swimming test (FST). In addition, CBD alleviated the neuroinflammation and inhibited the activation of microglia in mouse brain. In vitro, CBD significantly inhibited the activation of BV-2 microglia cells. Both in vivo and in vitro experiments confirmed that CBD could inhibit NF-κB expression. Conclusions:LPS could induce the activation of BV-2 microglia cells and the expression of inflammatory factors in mouse brain accompanied with abnormal behaviors. CBD could inhibit the activation of microglia, alleviate the neuroinflammation in different regions of mouse brain and improve behavioral performance.

Background Air pollutants PM_2.5 and its adsorbed metal elements are important factors affecting public health. Objective To explore the distribution characteristics and sources of metal elements in atmospheric PM_2.5 in Lanzhou from 2019 to 2020, and to assess the health risks of metal elements to different groups of residents through inhalation. Methods From January 2019 to December 2020 in two districts of Lanzhou City (Chengguan District and Xigu District), regular PM_2.5 and metal elements [antimony (Sb), aluminum (Al), arsenic (As), beryllium (Be), cadmium (Cd), chromium (Cr), mercury (Hg), lead (Pb), manganese (Mn), nickel (Ni), selenium (Se), and thallium (Tl)] were regularly monitored, and their concentrations were described by the median (M) and 25^th and 75^th percentiles (P₂₅, P₇₅) as not following a normal distribution (because the detection rates of the five elements Be, Cr, Hg, Ni, and Se were less than 70%, the five elements were not included in subsequent analysis), and then compared with the secondary concentration limits in the Ambient Air Quality Standards (GB 3095-2012). The differences between the medians of the two groups were compared by the Mann-Whitney U rank sum test, and the differences among the medians of multiple groups were compared by the Kruskal-Wallis H rank sum test; the enrichment factor (EF) method and principal component analysis were used to evaluate the pollution degree of the metals and their sources; the health risks of five non-carcinogenic metals (Sb, Al, Pb, Mn, and Tl) and two carcinogenic metals (As and Cd) in PM_2.5 were evaluated by hazard index (HI) and hazard quotient (HQ) using the incremental lifetime cancer risk (LCR) model and the non-carcinogenic risk assessment model, respectively. Results The PM_2.5 concentrations [M (P₂₅, P₇₅)] in Lanzhou City were 38.50 (26.00, 65.00) and 41.00 (29.00, 63.10) μg·m⁻³ in 2019 and 2020, respectively, and the difference was not statistically significant (Z=−0.989, P > 0.05). The average levels of the metal elements from high to low were: Al > Pb > Mn > As > Cd > Sb > Tl, and the annual average concentration of each metal element in 2019 was higher than that in 2020 (P<0.05). The M ( P₂₅, P₇₅) of PM_2.5 concentrations in Chengguan and Xigu districts were 52.98 (17.00, 61.00) and 55.40 (17.00, 67.00) μg·m⁻³, respectively, with no statistically significant differences (P<0.05); the concentrations of Sb and Al in Chengguan District were lower than those in Xigu District (P<0.05), and the concentrations of other metal elements were not different between the two areas (P>0.05). There were seasonal differences in the concentrations of PM_2.5 and seven metal elements in Lanzhou City (except P_Al=0.007, the other P_s < 0.001). The results of the enrichment factor method showed that the EF values of the six metals (Sb, Al, As, Cd, Pb and Tl) were all greater than 1. Among them, except As, the EF values of other metal elements were all greater than 10, and the EF values of Al and Cd were both greater than 100. The results of principal component analysis showed that the variance contributions of the three principal components were 45.61%, 24.22%, and 14.42%, and the cumulative contribution reached 84.25%. The principal component 1 included Pb, As, Cd, and Sb, the principal component 2 included Al and Mn, and the principal component 3 contained Tl. The non-carcinogenic risks of the five metals were, in descending order, Al > Mn > Pb > Tl > Sb, among which the HQ values of the remaining four metals were less than 1 for adults and children, except the HQ value of Al for adults, which was greater than 1. The ILC values of carcinogenic metal As for adult males, adult females, and children were 2.68×10⁻⁵, 2.51×10⁻⁵, and 1.45×10⁻⁵, respectively; the ILC values of carcinogenic metal Cd for adult males, adult females, and children were 1.53×10⁻⁶, 1.43×10⁻⁶, and 8.26×10⁻⁷, respectively. Conclusion There is pollution of atmospheric PM_2.5 and its adsorbed metal elements in Lanzhou. As and Cd elements may pose potential carcinogenic risks to the residents.

Background Polycyclic aromatic hydrocarbons (PAHs) are one of the most widely distributed and harmful organic pollutants in the atmosphere. Objective To investigate the distribution characteristics and composition sources of PAHs in the atmosphere of two districts of Lanzhou from 2019 to 2020 and evaluate the health risks of PAHs via inhalation to different populations. Methods The PAHs concentrations in two urban areas (Chengguan District and Xigu District) of Lanzhou City from January 2019 to December 2020 were regularly monitored. Mann-Whitney U test was used to compare differences in target pollutant concentrations between the two areas. Diagnostic ratio method and principal component analysis were adopted for source identification. Incremental lifetime cancer risk (ILCR) model was applied to evaluate the health risks of PAHs. Results The M (P₂₅, P₇₅) PAHs concentrations in Chengguan District and Xigu District were 24.04 (14.59, 41.81) ng·m⁻³ and 25.97 (18.59, 42.56) ng·m⁻³, respectively, with no significant difference (Z=−0.970, P>0.05). As to seasonal distribution, most PAHs monomer concentrations in Chengguan District were higher than those in Xigu District in summer, and the concentrations of benzo[a]anthracene and benzo[g,h,i]perylene in Chengguan District were also higher than those in Xigu District in spring and autumn (P<0.05), but there were no significant differences in PAHs monomer concentrations between the two urban areas in winter (P>0.05). Ring number of PAHs exhibited seasonal fluctuations. In winter and spring, the highest proportions in Chengguan District and Xigu District were both 4-ring PAHs (37.32%-41.73%, 35.20%-39.66%), and in summer and autumn, the highest proportions were both 2- and 3-ring PAHs (39.38%-49.54%, 47.17%-51.23%). The results of diagnostic ratio method revealed mixed atmospheric PAHs sources in the two urban areas, including fossil fuel, coal, and biomass combustion. The results of principal component analysis showed that the cumulative contribution rate of the four principal component factors reached 79.54%. Principal component 1 included phenanthrene, anthracene, fluoranthene, pyrene, chrysene, benzo[a]anthracene, benzo[b]fluoranthene, benzo[k]fluoranthene, and benzo[a]pyrene; principal component 2 included acenaphthene and dibenzo[a,h]anthracene; principal component 3 was fluorene; principal component 4 was naphthalene. The results of health risk assessment showed that the ILCR values of adult males, adult females, and children in Chengguan District were 2.30×10⁻⁶, 2.16×10⁻⁶, and 1.73×10⁻⁶, respectively; and those in Xigu District were 1.58×10⁻⁶, 1.48×10⁻⁶, and 1.19×10⁻⁶, respectively; all were greater than 10⁻⁶. Conclusion PAHs pollution exists in the atmosphere of the two urban areas of Lanzhou City, mainly comes from mixed sources of fossil fuels, coal, and biomass burning, and may pose potential carcinogenic risks to the population.

Organic carbonates (OCs) are a class of compounds featured by a carbonyl flanked by two alkoxy/aryloxy groups. They exist in either linear or cyclic forms, of which the majority encountered in nature adopt a pentacyclic structure. However, the enzymatic basis for pentacyclic carbonate ring formation remains elusive. Here, we reported that a four-protein metabolon (AlmUII-UV) assembled by a small peptide protein (AlmUV) appends a reactive

Objective:To summarize the clinical nursing practice of mitral valvuloplasty combined with coronary artery bypass grafting in a patient with hemophilia A who suffered acute mitral valve tendon rupture and coronary heart disease.Methods:Individualized care plans were developed for patients with hemophilia A who need mitral valvuloplasty and coronary artery bypass grafting. Nursing measures were conducted from anticoagulatiaon care, bleeding care, hypoxemia care, multimodal analgesic care, psychological care, discharge guidance and discharge follow-up.Results:After multidisciplinary comprehensive treatment and standardized, individualized nursing, the patient recovered well and discharged after surgery on the 13th day.Conclusions:Implement personalized anticoagulation programs and care for this patient's individual situation to ensure the patient's smooth perioperative period Effective implementation of various postoperative care measures to promote early recovery of patients and reduce the pain caused by the disease during their hospitalization, also to improve the patient's quality of life.

Objective:To explore the value of FilmArray meningitis/encephalitis(ME)Panel in etiological diagnosis of infection in central nervous system(CNS) in Chinese children.Methods:Cerebrospinal fluid(CSF)obtained through lumbar puncture was collected from 145 patients with suspected CNS infection at Shanghai Children′s Medical Center Affiliated to Shanghai Jiaotong University School of Medicine from March 2019 to November 2019.All specimens were cultured simultaneously, which were detected by FilmArray ME Panel, and the results of cerebrospinal fluid culture and FilmArray ME Panel were compared.Results:Among 145 patients with suspected CNS infection, three samples were found to be positive after cerebrospinal fluid culture, and the positive rate was 2.1%(3/145). For the FilmArray ME Panel, 30 specimens were found to be positive, with a positive rate of 20.7%(30/145), and the difference of positive rate between the two methods was statistically significant( χ2=24.927, P<0.05). Among the samples FilmArray ME Panel tested positive with pathogen, 26 specimens were positive with virus making up 17.9%(26/145)and enterovirus(15.2%)was the primary pathogen.In addition, of the 142 specimens cerebrospinal fluid culture negative, 28 samples were tested positive by the FilmArray ME Panel, accounting for 19.7%(28/142). Conclusion:FilmArray ME Panel has the characteristics with high positive rate and could be time-saving.Meanwhile, FilmArray ME Panel has significant advantage in the detection of virus and improves the positive detection rate of virus.

The aim of this study was to investigate the effect of transmembrane 9 superfamily protein member 2 (TM9SF2) in proliferation and migration of triple negative breast cancer cell line MDA-MB-231.The expression of TM9SF2 in triple negative breast cancer cell line MDA-MB-231 and nontumorigenic mammary epithelial cell line MCF-10A were measured by Western blot. MDA-MB-231 cells were treated with siRNA-TM9SF2 to knock-down the expression of TM9SF2. The effect of silencing TM9SF2 was measured with Western blot.The proliferation of cells was tested by MTS,and the migration was measured with Transwell and wound-healing assay.Proteins related to proliferation (PI3K,AKT,SRC and ERK) and migration (Snail,Slug and N-cadherin) were measured with Western blot.Protein expressions of TM9SF2 was better improved in triple negative breast cancer MDA-MB-231 cell line than MCF-10A.Compared with the control group, the siRNA-TM9SF2 infected group had lower expressions of PI3K, Snail, Slug and N-cadherin, and at the same time phosphorylation of AKT was decreased. The results suggest TM9SF2 can promote the proliferation and metastasis of triple negative breast cancer MDA-MB-231 cell line.