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 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.

Objective: To compare the overall survival (OS), progression-free survival (PFS) and brain metastasis free survival (BMFS) between the chemo-radiotherapy and surgical treatment for patients with limited stage small cell lung cancer (LS-SCLC). Methods: Clinical data of 69 patients diagnosed with LS-SCLC undergoing surgery in Zhejiang Cancer Hospital between 2000 and 2016 were collected. According to T, N stage, treatment duration, age, gender and whether or not prophylactic cranial irradiation (PCI), 69 patients of 503 LS-SCLC patients who underwent standard radiochemotherapy were assigned into the radiochemotherapy group by using the pair-matched case-control method. Results: Among 138 patients, 69 cases were allocated into the surgery group (24 cases of stage Ⅰ, 14 cases of stage Ⅱ and 31 cases of stage Ⅲ) and 69 cases in the radiochemotherapy group (24 cases of stage Ⅰ, 14 cases of stage Ⅱ and 31 cases of stage Ⅲ). The median OS time was 37.1 months (95%CI: 24.1-50.2 months) in surgery group and 45.0 months (95%CI: 15.8-74.2 months) in the radiochemotherapy group. The 2-and 5-year OS rates were 60% and 45% in the surgery group, and 64% and 45% in the radiochemotherapy group (P=0.846). The median PFS time was 27.1 months (95%CI: 0.00-60.3 months) in the surgery group and 36.2 months (95%CI: 20.9-51.4 months) in the radiochemotherapy group. The 2-and 5-year PFS rates were 52%, and 38% in the surgery group, and 56% and 40% in the chemo-radiotherapy group (P=0.610). The 2-and 5-year BMFS rates were 81% and 76% in the surgery group, and 84% and 80% in the radiochemotherapy group (P=0.774). The 5-year OS rate (62% vs. 40%, P=0.038) and 5-year PFS rate (80% vs.40%, P=0.048) for patients with stage Ⅰ LS-SCLC in the surgery group were significantly higher than those in the radiochemotherapy group. However, the 5-year BMFS rate in patients with stage Ⅰ LS-SCLC did not significantly differ between two groups (92% vs.95%, P=0.816). The 5-year OS rate (41% vs.51%, P=0.946), 5-year PFS rate (65% vs.42%, P=0.280) and 5-year BMFS rate (75% vs.78%, P=0.720) for stage Ⅱ SCLC did not significantly differ between two groups. As for stage Ⅲ SCLC patients, the OS rate (25% vs.48%, P=0.220), 5-year PFS rate (28% vs.36%, P=0.333) and 5-year BMFS rate (76% vs. 74%, P=0.84) did not significantly differ between two groups. Conclusions Surgical treatment can bring survival benefits to patients with stage Ⅰ LS-SCLC. The survival prognosis of stage Ⅱ patients is equivalent between two groups. Patients with stage Ⅲ LS-SCLC receiving radiochemotherapy obtain better survival trend compared with those undergoing surgery. The conclusion remains to be validated by studies with larger sample size or prospective investigations.

SARS-CoV-2 infection causes complicated clinical manifestations with variable multi-organ injuries, however, the underlying mechanism, in particular immune responses in different organs, remains elusive. In this study, comprehensive transcriptomic alterations of 14 tissues from rhesus macaque infected with SARS-CoV-2 were analyzed. Compared to normal controls, SARS-CoV-2 infection resulted in dysregulation of genes involving diverse functions in various examined tissues/organs, with drastic transcriptomic changes in cerebral cortex and right ventricle. Intriguingly, cerebral cortex exhibited a hyperinflammatory state evidenced by significant upregulation of inflammation response-related genes. Meanwhile, expressions of coagulation, angiogenesis and fibrosis factors were also up-regulated in cerebral cortex. Based on our findings, neuropilin 1 (NRP1), a receptor of SARS-CoV-2, was significantly elevated in cerebral cortex post infection, accompanied by active immune response releasing inflammatory factors and signal transmission among tissues, which enhanced infection of the central nervous system (CNS) in a positive feedback way, leading to viral encephalitis. Overall, our study depicts a multi-tissue/organ transcriptomic landscapes of rhesus macaque with early infection of SARS-CoV-2, and provides important insights into the mechanistic basis for COVID-19-associated clinical complications.
Animals ; COVID-19/genetics* ; Macaca mulatta ; SARS-CoV-2/genetics* ; Transcriptome