Objective To observe the efficacy of monosialotetrahexosy lganglioside (GM1) in the treatment of vascular cognitive impairment (VCI). Methods Sixty patients with VCI were randomly divided into either a treatment group or a control group. The patients in the control group received conventional treatment and those in the treatment control group were treated with conventional treatment plus continuous intravenous infusion of GM1 (80 mg/d) for 2 weeks. The efficacy was evaluated by the Montreal Cognitive Assessment (MoCA) before and after the treatment, and the safety of the treatment was observed. Results After 2 weeks of treatment, the MoCA scores were significantly higher than those before treatment in both groups (all P ＜ 0. 05). The MoCA scores of the treatment group were significantly higher than those of the control group (20. 82 ± 1. 96 vs. 19. 61 ±2. 02, t =2. 315, P =0. 023). No obvious adverse reactions were found. Conclusions The efficacy of GM1 is positive in the treatment of vascular cognitive impairment, and there is no obvious adverse reactions. It is worthy of using widely in clinical practice.

Objective To explore whether Helq deletion affect the pluripotency of stem cells. Methods Helq knockout embryonic stem cells were obtained by CRISPR?Cas9 gene editing technique. Results The results of immunoflu?orescence analysis showed that the expression of Oct4 and Nanog had no obvious difference to that of the control cells. The Helq-/ - embryonic stem cells could produce viable pups by tetraploid complementation, indicating that their pluripotency was not affected. Meanwhile, we found that day 2 epiblast?like cells also were obtained through differentiation of the Helq-/ - embryonic stem cells in vitro. Immunostaining and real?time PCR analysis showed that the gene expression of Helq-/ - epiblast cells were similar to the wild type cells. Conclusions Taken together, it is proved that the genomic in?stability caused by Helq deletion does not affect the pluripotency of pluripotent stem cells.

BACKGROUND: Memory loss is the main presentation during the earlier stage of radiative eneephalopathy, and it was reported that ganglioside (GM1) played important role in neural rehabilitation, particular in the improvement of memory.OBJECTIVE: To study the improving effect of GM1 on spatial learning and memory retardation in rats following radiative encepholopathy. DESIGN: Randomized control and comparative observing study based on the experimental animals.SETTING: Department of Neurology and Department of Radiation of Second Affiliated Hospital of Sun Yat-sen University.MATERIALS: This study was carried out at the Laboratory of the Second Hospital Affiliated to Sun Yat-sen University between March 2001 and May 2002. Tctally 80 SD rats were randomly selected and divided into control group, GM1 treatment group, physiological saline group and non-intervention group with 20 rats in each group.INTERVENTIONS: Rats in GM1 group, physiological saline group and non-intervention group subjected to head 60Coγ irradiation of 7Gy each time after anesthesia, once a day for consecutive 6 days, and the total dosage was 42Gy while rats in control group did not receive irradiation after anesthesia. Rats in GM1 and physiological saline(PS) group were given intraperitoneal injection of GM1 and physiological saline of 30 mg/kg respectively at 1 hour after each time of radiation, once a day for consecutive 6 days but not in control group and non-intervention group. Evaluation: ①After irradiation, morris water labyrinth navigation test was used to assess the capability of learning and memory of rats by the time for reaching platform (latency); ② Spatial searching test was used to detect their spatial memory after learning how to reach the platform by recording the way of rats searching the platform in 120 s and calculating the percentage of swimming distance in platform quadrant in the total distance; ③ After labyrinth test, brains were taken out of the rats in GM1 group, PS group and non-intervention group for observing the histological and pathological changes in rat brains.RESULTS: ① The latency become stable form onset of the 4th day in each group. On the 5th day, the searching platform latency in GM1 group was(13.6±1.4) s, shorter than(17.1±2.9) s of PS group and [(15.8±2.2) s, (P＜0.05)] of non-intervention group; ② Rats in GM 1 and control group were found capable of searching platform according to their spatial memory, presented by swimming trail most located in platform quadrant while rats in PS and non-intervention groups were found mostly swimming around the pool with moving trails distributed randomly. The percentage of swimming distance in platform quadrant was found higher in GM 1 treatment group than in the PS group and non-intervention group, but lower than that in the control group; ③ Histological examination revealed slight neuronal degeneration in PS group, part of which was changes of vacuolar degeneration with cell shrank, chromosome concentrated and nuclei gathered aside, and the number of astrocytes also decreased; the pathological changes in non-intervention group and PS group were similar; in GM1 group, part neurons became smaller with peripalsm turning red but the pathological changes, such as the number of cells,neuclei shrank and gathered aside, and vacuolar changes were less than those of the former two groups.ONCLUSION: Radiative encephalopathy would result in obvious learning nd memory impairments in rats but histological and pathological changes due o brain radiation injury can be attenuated with the treatment of GM1, implying that GM1 may play important role in the improvement of radiation-induced spatial learning and memory loss.

Objective: To explore the mechanism of abnormal expression of microRNA155 (miR155) in myeloma drug-resistance to probe the possibility of inhibiting miR155 expression to restore chemotherapy sensitivity and its molecular mechanism in drug-resistant myeloma cells. Methods: Drug-resistant myeloma cell-line RPMI8226/DOX was established by culturing RPMI8226 cells with continuous low concentration and intermittent gradually increasing concentration of doxorubicin in vitro; The levels of miR155 mRNA were measured by qRT-PCR, and both proteins FOXO3a and BCL-2 expressions were detected by Western blot in cell-lines RPMI8226/S and RPMI8226/Dox. RPMI8226/DOX cells were transfected by miR155 inhibitor and mimic using gene transfer method, and then CCK-8 was used to measure proliferation and inhibition ratio, the changes of miR155 expression were detected by RT-PCR. Proteins FOXO3a and BCL-2 were detected by Western blot. Results: Comparing with RPMI8226 cells, the level of miR155 mRNA was obviously up-regulated with the relative expression of 26.860±2.340, together with increased expression of Bcl-2 protein but decreased expression of FOXO3a in RPMI8226/DOX cells. After 72 h treatment with miR155 inhibitor, the inhibition rate of transfection was 64.57%, miR155 expression decreased sharply, the level of FOXO3a expression was upregulated while BCL-2 expression decreased, chemotherapy sensitivity was restored on cell-line RPMI8226/DOX with reversed drug-resistance ratio of 2.518. Conclusions The abnormal expression of miR155 was closely associated with myeloma drug-resistance, targeting inhibition of miR155 expression could restore chemotherapy sensitivity by increasing FOXO3a expression in drug-resistant myeloma cells.

Objective: To explore the effect of heme oxygenase-1 (HO-1) on level of reactive oxygen species (ROS) induced by zinc oxide nanoparticles (ZnO-NPs) in Human umbilical vein endothelial cells line EA.hy926. Methods: The EA.hy926 cells in logarithmic growth phase were incubated with 0.0, 2.5, 5.0, 10.0 and 15.0 mg/L ZnO-NPs respectively. The ROS level, reflected by mean fluorescence intensity (MFI), was examined by flow cytometer after 4 hours exposure, the protein expression of HO-1 which was determined by Western Blot after exposed to ZnO-NPs for 24 hours. Cells incubated with 15.0 mg/L were set as the ZnO-NPs group; a blank control group was set at the same time. Cells were pretreated with HO-1 inhibitor zinc protoporphyrin (ZnPPIx) and HO-1 activator cobalt protoporphyrin (CoPPIx), they were classified as ZnPPIx group and CoPPIx group. 15 mg/L ZnO-NPs was chosen to conduct the experiment of HO-1 activation and inhibition. Cells were classified as ZnPPIX+ ZnO-NPs group and CoPPIx+ ZnO-NPs group after pretreated with 10 μmol/L ZnPPIx or CoPPIx for 1 h, added 15 mg/L ZnO-NPs to cell culture medium. In all groups ROS levels were detected after exposed to ZnO-NPs for 4 hours, the protein expression of HO-1 was detected after exposed to ZnO-NPs for 24 hours. Results: With the increased dose of ZnO-NPs, levels of ROS and HO-1 in EA.hy926 cells were clearly elevated (the MFI of 0.0, 2.5, 5.0, 10.0 and 15.0 mg/L ZnO-NPs incubated groups was 22 627.22±718.27, 24 726.47±568.52, 31 141.75±1 312.24, 39 824.82±4 774.74, 50 569.03±1 497.63 respectively, and HO-1 relative expression were 0.16±0.01, 0.19±0.02, 0.16±0.01, 0.23±0.02, 0.92±0.06 respectively). HO-1 expression in ZnPPIx pretreatment group decreased compared with ZnO-NPs group (1.05±0.05 vs. 1.12±0.01, P<0.05), meanwhile ROS level enhanced (62 683.95±2 589.59 vs. 53 654.53±2 229.01, P<0.05). However, CoPPIx pretreatment had higher HO-1 level and lower level of ROS compared with ZnO-NPs group (HO-1: 1.74±0.11 vs. 0.22±0.03, P<0.05; ROS: 32 845.04±993.48 vs. 53 654.53±2 229.01, P<0.05). Conclusions Exposure to ZnO-NPs significantly induced ROS generation in EA.hy926 cells in a dose-dependent manner. HO-1 regulated ZnO-NPs-induced oxidative stress.

Objective:To evaluate the effect of inhalation of sevoflurane during cardiopulmonary bypass (CPB) on early postoperative brain injury in the patients undergoing cardiac valve replacement.Methods:Forty-two American Society of Anesthesiaologists physical status Ⅱ or Ⅲ patients of either sex, aged 40-70 yr, weighing 47-86 kg, scheduled for elective single valve replacement under CPB, were divided into 3 groups ( n=14 each) using a random number table method: control group (group C), combined intravenous-inhalational anesthesia group (group CA) and sevoflurane group (group S). During CPB, propofol 4-6 mg·kg -1·h -1 was intravenously infused in group C, propofol 2-3 mg·kg -1·h -1 was intravenously infused, and 0.5 MAC sevoflurane was inhaled via the membrane oxygenator in group CA, and 1.0-1.5 MAC sevoflurane was inhaled via the membrane oxygenator in group S. The anesthesia and sedation index values were maintained at 40-60 during operation in the three groups.Blood samples were taken from arteries before anesthesia induction (T 1), at 30 min and 6 and 24 h after termination of CPB (T 2-4) for determination of plasma concentrations of neuron-specific enolase (NSE) and Tau protein. Results:Compared with group C, the plasma concentration of NSE was significantly decreased at T 2, 3, and plasma concentration of Tau protein was decreased at T 2-4 in group S, and the plasma concentration of Tau protein was decreased at T 2 in group CA ( P<0.05). Compared with group CA, the plasma concentration of NSE was significantly decreased at T 2, 3, and the plasma concentration of Tau protein was decreased at T 2-4 in group S ( P<0.05). Conclusions:Inhalation of sevoflurane during CPB can reduce early postoperative brain injury to a certain extent in the patients undergoing cardiac valve replacement.

Objective:To investigate the role of RNA m6A methylation in mediating cerebellar dysplasia through analyzing the phenotypes of the mouse cerebella and the expression of several key m6A regulators upon hypobaric hypoxia treatment.Methods:Five-day old C57/BL6 mice were exposed to hypobaric hypoxia for 9 days. The status of mouse cerebellar development was analyzed by comparing the body weights, brain weights and histological features. Immunostaining of cell-type-specific markers was performed to analyze the cerebellar morphology. Real-time PCR, Western blot and immunohistochemical staining were performed to detect the expression of key m6A regulators in the mouse cerebella.Results:Compared with the control, the body weights, brain weights and cerebellar volumes of hypobaric hypoxic mice were significantly reduced ( P<0.01). The expression of specific markers in different cells, including NeuN (mature neuron), Calbindin-D28K (Purkinje cell) and GFAP (astrocyte), was decreased in hypobaric hypoxic mouse cerebella ( P<0.01), accompanied with disorganized cellular structure. The expression of methyltransferase METTL3 was significantly down-regulated in the cerebella of hypobaric hypoxic mice ( P<0.05). Conclusions:Hypobaric hypoxia stimulation causes mouse cerebellar dysplasia, with structural abnormalities in mature granular neurons, Purkinje cells and astrocytes. Expression of METTL3 is decreased in hypobaric hypoxic mice cerebellum compared with that of normobaric normoxic mice, suggesting that its mediated RNA m6A methylation may play an important role in hypobaric hypoxia-induced mouse cerebellar dysplasia.

Objective To systematically summarize the published literature on the genetic variants associated with nonalcoholic fatty liver disease(NAFLD). Methods Literature from Web of Science,PubMed,and Embase between January 1980 and September 2022 was systematically searched.Meta-analyses of the genetic variants were conducted using at least five data sources.The epidemiologic credibility of the significant associations was graded using the Venice criteria. Results Based on literature screening,399 eligible studies were included,comprising 381 candidate gene association,16 genome-wide association,and 2 whole-exome sequencing studies.We identified 465 genetic variants in 173 genes in candidate gene association studies,and 25 genetic variants in 17 genes were included in the meta-analysis.The meta-analysis identified 11 variants in 10 genes that were significantly associated with NAFLD,with cumulative epidemiological evidence of an association graded as strong for two variants in two genes(HFE,TNF),moderate for four variants in three genes(TM6SF2,GCKR,and ADIPOQ),and weak for five variants in five genes(MBOAT7,PEMT,PNPLA3,LEPR,and MTHFR). Conclusion This study identified six variants in five genes that had moderate to strong evidence of an association with NAFLD,which may help understand the genetic architecture of NAFLD risk.

Objective:To discuss the differences in eosinophil(EOS)infiltration in cervical tissue and its relationship with cervical-related diseases,and to clarify the effect of EOS on the occurrence and development of cervical intraepithelial neoplasia(CIN)and cervical cancer.Methods:The clinical data of 256 patients with cervical diseases were collected and divided into cervical cancer group(n=46,including 26 cases of squamous cell carcinoma,15 cases of adenocarcinoma,and 5 cases of adenosquamous carcinoma),chronic cervicitis group(n=50),CIN stage Ⅰ group(n=50),CIN stage Ⅱ group(n=50),CIN stage Ⅲ group(n=30),and normal group(adjacent normal cervical tissue,n=30)based on their conditions.Colposcopy was used to observe the morphology of cervical tissue of the patients in various groups;thin-layer liquid-based cytology test(TCT)was used to observe the morphology of the cervical exfoliated cells in various groups;hybrid capture-chemiluminescence method was used to detect the human papillomavirus(HPV)infection in cervical tissue of the patients in various groups;HE staining was used to observe the pathomorphology of cervical tissue of the patients in various groups;Congo red staining was used to detect the numbers of EOS infiltration in cervical tissue of the patients in various groups;Pearson correlation analysis was used to analyze the correlation between the number of EOS infiltration and the malignancy degree of cervical cancer.Results:The cervical surface of the patients in normal group was smooth and pink,with uniformly distributed capillaries;the cervical surface of the patients in chronic cervicitis group showed red inflammatory changes,with some accompanied by Nabothian cysts and varying degrees of erosion and ulcers;the patients in CIN stage Ⅰ,CIN stage Ⅱ,and CIN stage Ⅲ groups showed epithelial ulcers,thickening,and irregular morphology,with mosaic and punctate vessels;the cervical surface of the patients in cervical cancer group showed raised areas with neoplasms and necrotic ulcers,and they were fragile and prone to bleeding.After acetic acid staining,no obvious changes of the patients in normal group were observed.The cervix of the patients in chronic cervicitis group showed slight white changes that lasted for a short time;in CIN stage Ⅰ,CIN stage Ⅱ,and CIN stage Ⅲ groups,irregular thin acetowhite epithelium with map-like borders was observed,with increasingly acetowhite reactions and larger areas as the stages advanced.The cervix of the patients in cervical cancer group showed thick acetowhite epithelium that lasted longer,with rigid and clear contours.After iodine staining,the cervix of the patients in normal group was brown,with uniform coloration;the cervix of the patients in chronic cervicitis group showed poor coloration in inflammatory lesion areas;the cervix of the patients in CIN stage Ⅰ group showed iodine coloration in metaplastic areas,while the cervix of the patients in CIN stage Ⅲ group showed poor coloration in larger lesion areas;the cervix of the patients in cervical cancer group showed irregular surfaces with cauliflower-like growth and no coloration after iodine staining,appearing orange-yellow or mustard yellow.The TCT observation results showed there were no heteromorphic cells and few inflammatory cells in cervical exfoliated cells of the patients in infiltration in normal group;there were numerous neutrophils and EOS in exfoliated cervical cells without heteromorphic cells in chronic cervicitis group.The heteromorphic binucleated cells with high nuclear-cytoplasmic ratios and deeply stained nuclei were observed in cervical exfoliated cells of the patients in CIN stage Ⅰ and CIN stage Ⅱ groups.More heteromorphic cells with high nuclear-cytoplasmic ratios and irregular nuclear membranes were showed in cervical exfoliated cells of the patients in CIN stage Ⅲ group.The cervical exfoliated cells of the patients in cervical cancer group showed large and prominent nucleoli,clustering into syncytial changes.Compared with normal group,the atypial of cervical exfoliated cells in CIN stage Ⅰ,CIN stage Ⅱ,CIN stage Ⅲ,and cervical cancer groups was increased.The hybrid capture-chemiluminescence results showed that compared with normal and chronic cervicitis groups,the numbers of HPV infection and TCT heteromorphic cells of the patients in CIN stage Ⅰ,CIN stage Ⅱ,and CIN stage Ⅲ groups were increased(P<0.05);compared with CIN stage Ⅰ,CIN stage Ⅱ,and CIN stage Ⅲ groups,the numbers of HPV infection and TCT heteromorphic cells of the patients in cervical cancer group were increased(P<0.05).The HE staining results showed normal cell morphology and structure in normal group,with infiltration of inflammation cells such as neutrophils,monocytes,macrophages,EOS,and lymphocytes;in chronic cervicitis group,the infiltration of inflammatory cells was increased;in CIN group,the cervical cells showed slightly larger nucleoli and heteromorphic cells,with inflammatory cells mainly distributing around the hetermomorphic cells;in cervical cancer group,the cervical cells showed large and deeply stained nucleoli with significant atypia,and the infiltration of inflammatory cells around the cancer cells was increased.Compared with normal group,the numbers of inflammatory cells and EOS infiltration in cervical tissue of the patients in chronic cervicitis group were increased(P<0.05),and the numbers of inflammatory cells and EOS infiltration of the patients in CIN group were increased(P<0.05);compared with chronic cervicitis group,the number of inflammatory cells and EOS infiltration of the patients in CIN group were decreased(P<0.05);compared with chronic cervicitis group and CIN group,the numbers of inflammatory cells and EOS infiltration of the patients in cervical cancer group were increased(P<0.05).The EOS in cervical cancer tissue was mainly distributed around the cancer nests;compared with CIN stage Ⅰ group,the numbers of EOS infiltration in CIN stage Ⅱ and CIN stage Ⅲ groups were increased(P<0.05);compared with CIN stage Ⅱ group,the number of EOS infiltration in CIN stage Ⅲ group was increased(P<0.05).The higher the malignancy degree of the tumor,the more EOS infiltration was observed,and the number of EOS infiltration was positively correlated with the invasion depth of cervical cancer(r=0.533 0,P<0.01).Conclusion:HPV infection and EOS infiltration play a role in promoting the and occurrence development of cervical precancerous lesions and cervical cancer.

OBJECTIVE: To explore the effect of zinc oxide nanoparticles(ZnO NPs) on the oxidative damage in human alveolar type Ⅱ epithelial cell line A549.METHODS: The A549 cells in logarithmic growth phase were incubated with ZnO NPs solution at dose of 0,10,20 and 40 mg/L as 4 dose groups.The levels of reactive oxygen species(ROS) were measured by flow cytometer after 4 hours of exposure.The malondialdehyde(MDA) content and super oxide dismutase(SOD) activity were measured by microplate reader after 8 hours of exposure.RESULTS: The ROS levels in A549 cells exposed to 10,20,40 mg/L ZnO NPs were significantly increased compared with control group(P<0.05).The level of ROS increased with the exposure dose of ZnO NPs in A549 cells(P<0.01).The activities of SOD in A549 cells exposed to 10,20,40 mg/L ZnO NPs were significantly decreased compared with control group(P<0.05).The level of MDA and the ratios of MDA/SOD increased compared with control group(P<0.05).The activity of SOD in A549 cells decreased with the increase of ZnO NPs exposure dose(P<0.01),and the level of MDA and the ratios of MDA/SOD increased with the increase of exposure(P<0.01).CONCLUSION: ZnO NPs could induce lipid peroxidation in A549 cells with a dose-effect relationship.