L007-0069 kills Staphylococcus aureus in high resistant phenotypes.

Staphylococcus aureus, a common gram-positive pathogenic bacterium, is a main cause of hospital infection. The prevalence rate of methicillin-resistant S. aureus (MRSA) has made its treatment difficult in recent decades. Moreover, S. aureus in the highly tolerant format of biofilm or persister often renders infections refractory. Thus, developing new active compounds against resistant S. aureus is urgently needed. In this study, by a high-throughput screening assay, we identified a small molecule, L007-0069, that exhibited strong and effective bactericidal activity against S. aureus and its high resistance patterns, such as biofilms and persisters, with a low probability of inducing resistance. By molecular dynamics and fluorescent probe analysis, mechanistic studies revealed that the bactericidal activity of L007-0069 was mainly mediated by membrane disruption and metabolic disorder induction. Furthermore, L007-0069 showed effective anti-MRSA effects in vivo in both a wound infection model and a peritonitis-sepsis model, with no detectable toxicity observed at the therapeutic dosage. In conclusion, L007-0069 has the potential to become an alternative for the treatment of highly resistant S. aureus-related infections.

Thickness, vessel density of retina and choroid on OCTA in young adults (18-24 years old).

PURPOSE: The purpose of this study was to quantify thickness, vessel density (VD) of retina and choroid in young adults (18-24 years old) using OCTA. METHODS: This observational, cross-sectional study included 154 eyes from 77 young myopic adults. En-face angiogram OCTA was performed on a 3.00 × 3.00 mm region centered on the macula. Automated thickness calculations and macular maps were measured. Spherical equivalent refraction (SER) and AL were examined to determine associations with thickness, vessel density (VD) of retina and choroid. RESULTS: A total of 148 healthy eyes from 77 young myopic adults (29 males and 48 females) with a mean age of 21.80 ± 1.32 years (range: 18-24 years) were included. The mean SER and AL were - 4.06 ± 2.26D and 25.25 ± 1.28 mm, respectively. The mean retinal thickness (RT, ILM-RPE layer) was 240.91 ± 13.36 µm, the retinal superficial (SVD) and deep vessel density (DVD) in fovea region were 18.35 ± 4.77% and 32.99 ± 6.01%, respectively. The foveal avascular zone (FAZ) area was 0.31 ± 0.10 mm2. The mean subfoveal choroidal thickness (SFCT) and choriocapillaris (CC) perfusion area were 232.16 ± 56.65 µm and 2.17 ± 0.10 mm2, respectively. By Pearson's correlation analysis, SER was revealed to be negatively correlated with RT (r = -0.180, p = 0.028) and DVD (r = -0.185, p = 0.025) in fovea region. SER was revealed to be positively correlated with RT in nasal (r = 0.224, p = 0.006) and inferior (r = 0.217, p = 0.008) regions. AL was revealed to be positively correlated with RT (r = 0.250, p = 0.002) and DVD (r = 0.284, p < 0.001) in fovea region. SER was revealed to be positively correlated with SFCT (r = 0.486, p < 0.001). AL was revealed to be negatively correlated with FAZ area (r = -0.232, p = 0.005) and SFCT (r = -0.407). RT was revealed to be negatively correlated with FAZ area (r = -0.645, p < 0.001). SER (r = -0.079), AL (r = 0.071) and SFCT (r = 0.089) did not correlate significantly with the CC perfusion area (all p > 0.05). CONCLUSION: Myopic eyes present increased RT, DVD and thinned SFCT in fovea, while no significant correlation could be found between SER, AL, SFCT and CC perfusion area. It may indicate that the SFCT thinning may be secondary to ocular elongation, while the CC perfusion area may be a factor independent of AL growth.

Study on antibacterial effect of halicin (SU3327) against Enterococcus faecalis and Enterococcus faecium.

Enterococci are important pathogens of nosocomial infections and are increasingly difficult to treat due to their intrinsic and acquired resistance to a range of antibiotics. Therefore, there is an urgent need to develop novel antibacterial agents, while drug repurposing is a promising approach to address this issue. Our study aimed to determine the antimicrobial efficacy of halicin against enterococci and found that the minimum inhibitory concentrations (MIC) of halicin against different strains of Enterococcus faecalis and Enterococcus faecium ranged from 4 to 8 µg/ml. In addition, the synergistic antibacterial effect between halicin and doxycycline (DOX) against Enterococcus was observed through the checkerboard method, and it was observed that halicin and DOX could significantly synergistically inhibit biofilm formation and eradicate preformed biofilms at sub-MICs. Moreover, the electron microscope results revealed that halicin could also disrupt the bacterial cell membrane at high concentrations. Furthermore, it is also confirmed that the combination of halicin and DOX has no significant cytotoxic effect on erythrocytes and other human-derived cells. In addition, the mouse subcutaneous model and H&E staining showed that the combination of halicin and DOX could effectively reduce the bacterial load and inflammatory infiltration without obvious side effects. In nutshell, these results demonstrate the potential of halicin in combination with DOX as a novel therapy against infections by Enterococcus.

Exosome-loaded injectable hydrogel for repairing bone defects around implants / 中国组织工程研究

BACKGROUND:A large number of studies have confirmed that exosomes can promote osteogenesis and vascularization.However,simple exosome therapy has problems such as poor targeting,and the content of loaded molecules cannot reach the therapeutic concentration. OBJECTIVE:To load exosomes into injectable gluconolactone-sodium alginate β-tricalcium phosphate-polyethylene glycol hydrogel,and observe the effect of the hydrogel on peri-implant bone defect in vivo and in vitro. METHODS:Exosomes were extracted from bone marrow mesenchymal stem cells and wrapped in injectable gluconolactone-sodium alginate β-tricalcium phosphate-polyethylene glycol hydrogel.(1)In vitro experiment:The hydrogel loaded with exosomes and the hydrogel without exosomes were cocultured with endothelial progenitor cells,and exosomes uptake experiment,tubule formation experiment,cell proliferation,migration ability,and angiogenic gene detection were carried out.(2)In vivo experiment:Twelve male New Zealand white rabbits were used to prepare two standard implant cavities and corresponding bone defects in the long axis of one femur.A hydrogel loaded with exosomes was implanted in the bone defect after an implant was implanted in a cavity at the proximal end of the implant(experimental group),and an unloaded exosome hydrogel was implanted in the bone defect after an implant was implanted in a cavity at the distal end of the implant(control group).At 3,6 and 9 weeks after operation,bone defects with implants were removed and stained with hematoxylin-eosin staining and Masson staining.Simultaneously,osteogenic and angiogenic genes were detected at 9 weeks after operation. RESULTS AND CONCLUSION:(1)In vitro experiment:Exosomes could enter endothelial progenitor cells.The proliferation,migration,angiogenesis and gene(CD31,vascular endothelial growth factor and basic fibroblast growth factor)expression of endothelial progenitor cells in the hydrogel-loaded group were higher than those in the hydrogel-unloaded group(P<0.05).(2)In vivo experiment:Hematoxylin-eosin staining and Masson staining showed that at 3 weeks after operation,only a small amount of new bone was found in the two groups,and the material was partially degraded.At 6 weeks after operation,the amount of new bone in the two groups increased,and a large amount of new bone was found in the experimental group,with obvious calcium deposition.At 9 weeks after operation,compared with the control group,a large number of bone trabeculae thicker than mature were found in the experimental group,calcium salt deposition was more obvious,and a large number of osteoblasts were found around the bone trabeculae.The protein expressions of CD31,vascular endothelial growth factor,basic fibroblast growth factor,bone morphogenetic protein 2,type I collagen and osteocalcin in the experimental group were higher than those in the control group at 9 weeks after operation(P<0.05).(3)The exosome-loaded gluconolactone-sodium alginate β-tricalcium phosphate-polyethylene glycol hydrogel could promote the proliferation,migration and angiogenic differentiation of endothelial progenitor cells and promote the repair and regeneration of bone defects around implants.

Preliminary study on high throughput screening small molecules targeting Gram-negative bacilli outer membrane protein BamA / 中华检验医学杂志

Objective:High-throughput screening to obtain small molecular compounds against Gram-negative bacilli by targeting BamA outer membrane protein.Methods:The sybyl-X2.1 software was used to perform high-throughput virtual screening of small molecular compounds in Chemdiv compound library based on the molecular docking. The top 150 hits by high-throughput screening were re-screened through in vitro biological experiments. The top 4 small molecules with obvious antibacterial activity were selected for in-depth molecular docking analysis, and the small molecule 8308-0401 with the highest docking score was selected for further experiments. The antibacterial effect of 8308-0401 combined with rifampicin was tested by checkerboard assay. Finally, the affinity between 8308-0401 and BamA was tested by plasma surface resonance assay. Results:The docking score of the top 150 hits calculated by high-throughput virtual screening had a mean value of 5.63. In vitro biological experiments showed that small molecules 8308-0401, 8365-1335, C066-2507 and L582-0346 exhibited strong antibacterial activity. Among those molecules, 8308-0401 showed the highest molecular docking score, and synergistic antibacterial activity against both types of strains and clinical isolates when combined with rifampicin. 8308-0401 has a strong affinity to BamA with binding a constant of 182 μmol/L. Conclusion:The small molecule 8308-0401 exerts antibacterial activity against Gram negative bacilli by targeting the outer membrane protein BamA.

Role of HO-1 in endotoxin-induced acute lung injury in mice: relationship with regulation of mitochondrial quality control / 中华麻醉学杂志

Objective:To evaluate the role of heme oxygenase-1 (HO-1) in endotoxin-induced acute lung injury (ALI) and the relationship with the regulation of mitochondrial quality control in mice.Methods:Clean-grade healthy male adult C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were selected.HO-1 inducible gene knockout mice (HO-1 -/-) were prepared based on CRISPER/Cas9-mediated EGE system, and HO-1 gene overexpression mice (HO-1 + /+ ) were prepared by transfection of HO-1 overexpressed adenovirus vector.The mice were divided into 2 groups ( n=6 each) using a random number table method: control group (group WT, group HO-1 -/-, group HO-1 + /+ ) and endotoxin-induced ALI group (group ALI, group HO-1 -/-+ ALI, group HO-1 + /+ + ALI). Lipopolysaccharide 15 mg/kg was injected through the tail vein to develop the model of endotoxin-induced ALI, and the equal volume of normal saline was given instead in each control group.The mice were sacrificed by bloodletting at 12 h after lipopolysaccharide or normal saline administration.The lung tissues were harvested for microscopic examination of the pathological changes which were scored, for determination of GSH and GSSG contents, for observation of the ultrastructure of mitochondria (with a transmission electron microscope) and survival within 12 h, for measurement of mitochondrial membrane potential (MMP) levels, and for determination of the expression of mitochondrial quality control-related proteins mitochondrial fusion protein 2 (Mfn2) and dynamin-related protein 1 (Drp1), peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α), nuclear respiratory factor 1 (NRF1), mitophagy marker protein PTEN-induced kinase 1 (PINK1) and E3 ubiquitin-protein ligase Parkin.The ratio of GSH/GSSG was calculated. Results:Compared with control group (group WT, group HO-1 + /+ and group HO-1 -/-), the 12-h survival rate and MMP were significantly decreased, the lung injury score was increased, GSH content and GSH/GSSG ratio were decreased, and the content of GSSG was increased in endotoxin-induced ALI groups (group ALI, group HO-1 + /+ + ALI and group HO-1 -/-+ ALI) ( P<0.05). Compared with group ALI, the 12-h survival rate and MMP were significantly decreased, the lung injury score was increased, the GSH content and GSH/GSSG ratio were decreased, the GSSG content was increased, and the expression of HO-1, Mfn2, PGC-1α, NRF1, PINK1 and Parkin was down-regulated, and Drp1 expression was up-regulated in group HO-1 -/-+ ALI, and 12-h survival rate and MMP were significantly increased, lung injury score was decreased, GSH content and GSH/GSSG ratio were increased, GSSG content was decreased, the expression of HO-1, Mfn2, PGC-1α, NRF1, PINK1 and Parkin was up-regulated, and the expression of Drp1 was down-regulated in group HO-1 + /+ + ALI ( P<0.05). Conclusions:HO-1 is involved in the process of endotoxin-induced ALI in mice, which is related to the regulation of mitochondrial quality control.

Relationship between SIRT3 and mitochondrial function in mice with endotoxin-induced lung injury / 中华麻醉学杂志

Objective:To evaluate the relationship between silent information regulator 2 homologue 3 (SIRT3) and mitochondrial function in mice with endotoxin-induced lung injury.Methods:Twenty clean-grade healthy adult male wild C57BL/6 (SIRT3 + /+ ) mice, 20 SIRT3 knockout (SIRT3 -/-) mice, weighing 20-25 g, aged 6-8 weeks, were studied.SIRT3 + /+ mice and SIRT3 -/- mice were divided into 4 groups ( n=5 each) according to the random number table method: blank control group (group C, group SIRT3 -/-C), endotoxin-induced lung injury group (group L, group SIRT3 -/-L), endotoxin-induced lung injury plus resveratrol group (group L+ R, group SIRT3 -/-L+ R), and resveratrol group (group R, group SIRT3 -/-R). Resveratrol 15 mg/kg was intraperitoneally injected once a day for 7 consecutive days in L+ R, R, SIRT3 -/-L+ R and SIRT3 -/-R groups, while the equal volume of normal saline was injected in the rest groups.Lipopolysaccharid 15 mg/kg was injected via the tail vein to develop a mouse model of endotoxin-induced lung injury at 30 min after resveratrol injection on 7th day, in L+ R and SIRT3 -/-L+ R groups and at the corresponding time points in L and SIRT3 -/-L groups, while the equal volume of normal saline was injected in the other groups.Blood samples were collected from the orbital venous plexus at 12 h after injection of normal saline or lipopolysaccharid for determination of serum total oxidation state (TOS) and total antioxidant state (TAS) levels by the xylenol orange method and ABTS colorimetric method, and the oxidative stress index (OSI) was calculated.After the mice were sacrificed, the lung tissues were taken for microscopic examination of the pathological changes which were scored and for determination of the mitochondrial membrane potential (MMP) (by JC-1 method), cellular oxygen consumption rate (OCR) (by the specific fluorescent probe method), and expression of SIRT3 (by Western blot). Results:Compared with group C or group SIRT3 -/-C, the lung injury score, serum TOS concentration and OSI were significantly increased, TAS concentration, MMP and OCR were decreased, and SIRT3 expression was down-regulated in L, L+ R, SIRT3 -/-L and SIRT3 -/-L+ R groups ( P<0.05). Compared with group L, the lung injury score, serum TOS concentration and OSI were significantly decreased, TAS concentration, MMP and OCR were increased, and SIRT3 expression was up-regulated in group L+ R, and lung injury score, serum TOS concentration and OSI were significantly increased, TAS concentration, MMP and OCR were decreased, and SIRT3 expression was down-regulated in group SIRT3 -/-L ( P<0.05). Compared with group L+ R, the lung injury score, serum TOS concentration and OSI were significantly increased, the TAS concentration, MMP and OCR were decreased, and the expression of SIRT3 was down-regulated in group SIRT3 -/- L+ R ( P<0.05). There was no significant difference in the indicators mentioned above between group SIRT3 -/-L+ R and group SIRT3 -/-L ( P>0.05). Conclusions:Down-regulation of SIRT3 expression can lead to impaired mitochondrial function, which is involved in the pathophysiological mechanism of endotoxin-induced lung injury.

A retrospective study of cisatracurium besilate-assisted treatment of severe acute pancreatitis complicated with intraabdominal hypertension / 中华麻醉学杂志

The clinical data of patients with severe acute pancreatitis complicated with intraabdominal hypertension or abdominal compartment syndrome admitted to our Department of Critical Care Medicine from January 1, 2018 to October 1, 2021 were collected and analyzed.Patients were divided into a conventional treatment group and conventional treatment plus IV infusion of cisatracurium besilate group (muscle relaxation group). A prediction model of treatment propensity score was developed for paired screening, with 31 cases in each group.The conventional treatment group adopted conventional basic treatment methods such as gastrointestinal decompression, spasmolysis and analgesia, fluid therapy, inhibition of gastric acid, suppression of parenzyme, nutritional support, mechanical ventilation, and enemata.In muscle relaxation group, cisatracurium besilate was intravenously infused on the basis of routine treatment with the initial dose of 0.15 mg/kg given to facilitate endotracheal intubation, followed by continuous intravenous infusion at 1-3 μg·kg -1·min -1, and the dose was adjusted according to the patient′s basic vital signs and clinical effects.The primary outcome was survival rate.Secondary outcome measures were changes in intraabdominal pressure, oxygenation index, the number of defecation, volume of defecation, and urination volume before treatment and on 7, 14 and 20 days of treatment.and the recovery time of bowel sounds, length of mechanical ventilation, time of intensive care unit treatment, and total hospitalization costs.Compared with conventional treatment group, the survival rate was significantly increased, the intraabdominal pressure was decreased on 7, 14 and 20 days of therapy, the oxygenation index was increased, the number of defecation and volume of defecation were increased on 7 and 14 days of therapy, urinary volume was increased before treatment and on day 7 of therapy, the recovery time of intestinal sound was significantly shortened ( P<0.05), and no significant change was found in urinary volume on days 14 and 20 of therapy, length of ventilation, time of intensive care unit treatment, and total hospitalization costs in muscle relaxation group ( P>0.05). In conclusion, cisatracurium besilate can improve oxygenation, promote the recovery of intestinal function and improve the survival rate when used to assist the treatment in the patients with severe acute pancreatitis complicated with intraabdominal hypertension.

Effects of transcutaneous electrical acupoint stimulation on mitochondrial quality control during endotoxin-induced acute lung injury in mice / 中华麻醉学杂志

Objective:To evaluate the effect of transcutaneous electrical acupoint stimulation (TEAS) on mitochondrial quality control during endotoxin-induced acute lung injury (ALI) in mice.Methods:Twenty-four clean-grade healthy male C57BL/6J mice, aged 4-6 weeks, weighing 15-20 g, were divided into 4 groups ( n=6 each) according to the random number table method: control group (group C), endotoxin-induced ALI group (group L-ALI), endotoxin-induced ALI plus acupoint electroacupuncture group (group L-ALI+ EA), and endotoxin-induced ALI plus non-acupoint electroacupuncture group (group L-ALI+ SEA). Lipopolysaccharide (LPS) 15 mg/kg was injected via the caudal vein to develop the model of endotoxin-induced ALI in anesthetized mice.In group L-ALI+ EA, at 5 days before LPS injection, bilateral Zusanli and Feishu acupoints were stimulated with an electric stimulator for 30 min each time at a voltage of 1-2 mA and a frequency of 2/15 Hz until the end of the experiment.In group L-ALI+ SEA, stimulation was performed at the points 0.5 cm lateral to the acupoints of bilateral Zusanli and Feishu non-meridian and non-acupoint sites using the shallow puncture method, and the other treatment methods were the same as those previously described in group EA.Group C received no treatment.The mice were sacrificed by euthanasia at 12 h after LPS administration, and lung tissues were obtained for microscopic examination of the pathological changes (with a light microscope) and structure and morphology of mitochondria (with a transmission electron microscope) and for determination of the levels of reactive oxygen species (ROS) and mitochondrial DNA (mtDNA) and contents of glutathione (GSH) and glutathione oxidized (GSSG). The GSH/GSSG ratio was calculated.The expression of mitochondrial fusion proteins mitofusin 1 (Mfn1), Mfn2, optic atrophy1 (OPA1), dynamin-related protein 1 (Drp1), fission protein 1 (Fis1), peroxisome-proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor-1 (NRF1), NRF2, PTEN-induced putative protein kinase 1 (PINK1) and the E3 ubiquitin ligase (Parkin) was determined by Western blot. Results:Compared with group C, the level of ROS and contents of GSSG and mtDNA were significantly increased, GSH content and GSH/GSSG ratio were decreased, the expression of Mfn1, Mfn2, OPA1, NRF1, NRF2 and PGC-1α was down-regulated, and the expression of Drp1, Fis1, PINK1 and Parkin was up-regulated in L-ALI, L-ALI+ EA and L-ALI+ SEA groups ( P<0.05). Compared with group L-ALI, the level of ROS and contents of GSSG and mtDNA were significantly decreased, GSH content and GSH/GSSG ratio were increased, the expression of Mfn1, Mfn2, OPA1, NRF1, NRF2 and PGC-1α was up-regulated, and the expression of Drp1, Fis1, PINK1 and Parkin was down-regulated in group L-ALI+ EA ( P<0.05). Compared with group L-ALI+ EA, the level of ROS and contents of GSSG and mtDNA were significantly increased, GSH content and GSH/GSSG ratio were decreased, the expression of Mfn1, Mfn2, OPA1, NRF1, NRF2 and PGC-1α was down-regulated, and the expression of Drp1, Fis1, PINK1 and Parkin was up-regulated in group L-ALI+ SEA ( P<0.05). Conclusions:TEAS can reduce endotoxin-induced ALI probably through regulating mitochondrial quality control in mice.

Etiological analysis of the immunosuppressed children with severe pneumonia by metagenomics next generation sequencing / 中国小儿急救医学

Objective:To explore the application of metagenomics next generation sequencing(mNGS)in the immunosuppressed children with severe pneumonia and to understand the distribution of pathogens in order to provide reference for early prevention and treatment.Methods:We performed a retrospective analysis of the immunosuppressed children with severe pneumonia who had mNGS reports admitted to PICU according with the enrollment condition from July 2019 to July 2020.The records included general clinical data, traditional detection method report and mNGS results.We evaluated the consistency of the mNGS with the clinical microbiology reports and clinical judgment.Results:Twenty-three patients were enrolled, 15 were male and 8 were female, aging from 28 days to 10 years old, with an average age of(3.67±3.20)years old.Seven cases were cured, 2 were improved, and 14 died.A total of 23 samples were obtained, including 21 blood specimens and 2 bronchoal-veolar lavage fluid specimens.Among the 23 cases, 5 were single infected and 15 were mixed infected.Fungi were detected in 15 cases(65.22%), including 12 cases of Pneumocystis jirovecii, 2 cases of Aspergillus fumigatus and 2 cases of Candida albicans.Virus were detected in 14 cases(60.87%), including cytomegalovirus(CMV) in 10 cases(8 cases with pneumocystis infection), Herpes virus in 3 cases and fine ring virus in 2 cases(1 case with herpes virus infection). Bacteria were detected in 10 cases, including 3 cases of Acinetobacter, 1 case of Klebsiella pneumoniae, 1 case of Stenotrophomonas maltophilia, 1 case of Pinocytogenes, 4 cases of Staphylococcus and 1 case of Bacillus licheniformis.There were Mycoplasma in 3 cases with mixed infection.The positive rate and coincidence rate of mNGS were significantly higher than that of the traditional test group( P<0.05). A total of 19 cases were treated with hormone or immunosuppressive agents, and 17 cases were treated for 1 to 6 months when severe pneumonia occurred. Conclusion:Most immunosuppressed children with severe pneumonia are mixed infection.The common pathogens are Pneumocystis jirovecii and CMV.The use of mNGS can significantly improve the pathogen detection rate, effectively guiding the treatment.

Transcranial direct current stimulation combined with functional electrical stimulation-assisted cycling promote the recovery of lower extremity motor function early after stroke / 中华物理医学与康复杂志

Objective:To observe the effect of combining transcranial direct current stimulation (tDCS) with functional electrical stimulation-assisted cycling (FES-cycling) on lower limb motor function early after a stroke.Methods:Thirty-seven survivors of a recent stroke were divided into a tDCS treatment group ( n=18) and a pseudo-stimulation group ( n=19). While receiving routine rehabilitation training and clinical drug treatment, the tDCS treatment group also cycled in response to functional electrical stimulation while simultaneously receiving tDCS anode stimulation of the motor cortex M1 area. The pseudo-stimulation group followed the same protocol but with the tDCS stimulation inactivated. Both groups were treated for 20min daily, 5 days weekly for 4 weeks. Before and after the 4 weeks of treatment, the lower limb motor function, walking ability and ability in the activities of daily living of both groups were evaluated using the Fugl-Meyer assessment scale for the lower extremities (FMA-LE), the timed up and go test (TUGT) and the modified Barthel index (MBI) respectively. Transcranial magnetic stimulation was used to detect each subject′s cerebral cortex motor threshold (CMT) , cortical latency (CL) and central motor conduction time (CMCT) as well as the amplitude (Amp) of the motor evoked potential of the lower limb primary motor cortex (M1 area). Results:After 4 weeks of treatment, the average FMA-LE and MBI scores and TUGT times of the two groups had improved significantly compared with those before treatment. The average FMA-LE score and TUGT time of the tDCS group were significantly better than those of the pseudo-stimulation group. The average CMT, CL and CMCT in both groups were significantly lower than those before the intervention, while the average Amp had increased significantly, but there were significant differences in the average CMT, Amp, CL and CMCT between the two groups after the 4 weeks of treatment.Conclusions:Transcranial direct current stimulation combined with cycling assisted by functional electrical stimulation can effectively stimulate excitability in the motor cortex soon after a stroke. That should promote the recovery of nerve activity and lower limb function.

Correlation between altered levels of neurotransmitters in the frontal lobe and hippocampus and behavioral abnormalities in a Clock mutant mice modeling bipolar manic disorder / 中华医学遗传学杂志

OBJECTIVE@#To explore the correlation between altered levels of neurotransmitters in the frontal lobe and hippocampus and behavioral abnormalities in a Clock variant mice modeling bipolar disorder manic disorder.@*METHODS@#Open field test and Elevated plus-maze test were carried out on the Clock mutant and wild-type control groups. The frontal lobe and hippocampus of Clock mutant mice and controls were dissected, and neurotransmitters in tissue extracts were analyzed by high-performance liquid chromatography and mass spectrometry. The concentration of neurotransmitters and behavioral indicators were assessed by t test and Pearson correlation analysis using SPSS 22.0.@*RESULTS@#The Clock mutant mice showed a significant increase in activity, albeit with no difference in the level of anxiety from the wild-type controls, which suggested that the Clock mutant mice can be used as a model for manic attack of bipolar disorder. Altered neurotransmitter levels were detected in the frontal and hippocampal regions, including elevated histamine in the left hippocampus, reduced histamine in the right hippocampus, reduced gamma-aminobutyric acid (GABA) in bilateral hippocampus, elevated dihydroxyphenylalanine (DOPA) in the left frontal lobe and reduced DOPA in the right hippocampus, and decreased glutamine in bilateral frontal lobes. The reduced glutamine in the left frontal lobe and GABA in the right hippocampus correlated with the increased activity of Clock mutant mice.@*CONCLUSION@#Clock mutant mice showed abnormal behavior with increased activity. Reduced glutamine in the left frontal lobe and GABA in the right hippocampus were correlated with increased activity.

Clinical analysis of senile epilepsy / 中华老年医学杂志

Objective To investigate the etiology,clinical characteristics,treatment and prognosis of senile epilepsy.Methods Clinical data of 121 senile patients with epilepsy admitted to the First Affiliated Hospital of Chongqing Medical University from July 2011 to February 2017 were retrospectively analyzed.Results Ninety-six cases (79.3％) had definite causes.The common causes included cerebrovascular disease (59.4％),intracranial tumors (13.5％),central nervous system infection (8.3％),craniocerebral trauma (5.2％) and acute metabolic or toxic factors (11.5％).Seventy-one cases (58.7％) suffered from partial seizure,29 cases (24.0％) suffered from generalized seizures,17 cases (14.1％) from status epilepticus,and 4 cases (3.3％) from seizures which can not be classified.Sodium valproate and levetiracetam were the most commonly used drugs.Clinical conditions in 82 cases (97.6％) were controlled effectively with drug therapy,and most (88.1％) were treated with only a single drug.Conclusions There are several underlying etiologies in the elderly patients with epilepsy.The most common cause is cerebrovascular diseases,followed by intracranial tumors,central nervous system infection and trauma.In addition,acute metabolic factors or toxic factors cannot be ignored.Partial seizure is the most common type of seizures,especially the complex partial seizure.Monotherapy can reduce seizures in most elder patients.

The molecular and electrophysiological mechanism of buyanghuanwu decoction in learning and memory ability of vascular dementia rats.

Buyanghuanwu Decoction (BYHWD), as a traditional Chinese medicine, has been developed to treat vascular dementia for hundreds of years, but the underlying mechanisms remain unknown. In this research, the protective effects of BYHWD on hippocampal neuron were examined in the rats of ischemia-reperfusion. Ischemia-reperfusion injury was induced by the four-vessel occlusion method and continued for 30 days. BYHWD (per 6.25g/kg/d) was orally given to rats twice each day for 30 days after ischemia-reperfusion, Nimodipine (per 10mg/kg/d) was orally given to rats twice each day for 30 days. In VD+BYHWD group rats, the neuronal injury in the hippocampal CA1 region was significantly less than that of VD group's. BYHWD of intragastric administration also markedly increased the expression of Extracellular signal-regulated kinase 2 (ERK2) and Calcium/calmodulin-dependent protein kinaseII (CaMKIIIy)in the CA1 region. Our results suggested that increased ERK2 and CaMKIIIy due to BYHWD may partially account for its effect of neuroprotection standing against ischemic injury in the hippocampal CA1 region, and participated in the rebuilding of synapse, strengthened the expression of LTP, promoted the ability recover of learning and memory in VD rats.

Therapeutic effect of autologous periosteum on the healing of tendon-bone interface in rabbit rotator cuff tear / 中国比较医学杂志

Objective To study the effect of autologous periosteum on the healing of tendon-bone interface in rabbit rotator cuff tear. Methods Supraspinatus tenotomy was performed on one side shoulder in 60 New Zealand white rabbits to establish the model of rotator cuff tear. The rabbits were randomly divided into two groups:the study group(used autologous periosteum to promote the suture fixation)and control group(simple suture fixation). The rabbits were sacrificed at 4,8 and 12 weeks postoperatively with twenty rabbits sacrificed each time. Tissue samples of the tendon-bone interface was taken for histological examination and biomechanical test was performed to assess the strength of tendon-bone junction. Results At 4 weeks the study group showed extensive inflammatory cell infiltration, and a small amount of chondrocytes and extracellular matrix. At 8 weeks, the study group showed a large amount of immature chondrocytes arranged rather regularly. At 12 weeks,the tendon bone junction appeared similar to normal. In the study group,both the tendon-bone connection and the arrangement of chondrocytes were significantly better than the control group. The result of biomechanical testing showed that the highest tendon load in the study group was significantly higher than the control group (P < 0.05). Conclusions The use of autologous periosteum as a patch to strengthen the repair of the rotator cuff tear can effectively promote the healing of tendon bone interface, shorten the rotator cuff healing time and has good biological properties. This method provides an experimental basis for clinical rotator cuff repair surgery.

Clinical significance of microrna-625 expression in esophageal squamous cell carcino-ma / 中国肿瘤临床

Objective: To analyze the correlation of miR-625 expression with clinicopathological characteristics in esophageal squa-mous cell carcinoma (ESCC) and to explore the effect of miR-625 on the migration and proliferation of ESCC cells. Methods:The expres-sion level of miR-625 was determined through real-time PCR in 86 paired human ESCC tissue specimens and tumor-adjacent normal esophageal tissue specimens, ESCC cell lines, and esophageal epithelial cell line. The associations of miR-625 expression with clinico-pathological characteristics and survival in ESCC patients were analyzed. Transwell and CCK-8 assays were performed to examine the effect of miR-625 expression on migration and proliferation of ESCC cells. Results:Compared with tumor-adjacent normal specimens, miR-625 was significantly downregulated in ESCC tissue specimens (P<0.05). MiR-625 expression was decreased in ESCC cell lines com-pared with human esophageal epithelial cell lines (P<0.05). Lower miR-625 expression was associated with poorer prognosis and sur-vival. The migration and proliferation abilities of ESCC cells were inhibited by miR-625 overexpression (P<0.05). Conclusion:MiR-625 acts as a tumor suppressor gene in the development and progression of ESCC, suggesting that miR-625 may serve as an efficient prog-nosis biomarker and a potential therapeutic target for ESCC.

PM2.5 induces VEGF expression and inflammatory responses by trans-activating AP-1 in human bronchial epithelial cells / 军事医学

Objective To explore the role of the transcriptional factor activator protein (AP)-1 in mediating vascular endothelial growth factor ( VEGF) expression in human bronchial epithelial cells exposed to PM 2.5.Methods Beas-2B cells was treated with PM2.5.Luciferase assay was used to detect the activation status of AP-1 and transcription of VEGF in the Beas-2B cells.The induced activation of c-Jun, ATF2 and VEGF expression was tested by Western blotting assay.Results PM2.5 induced transactivation of the transcriptional factor AP-1, accompanied by phosphorylation of the AP-1 components, c-Jun and ATF2 in Beas-2B cells.Moreover, when AP-1 activation was inhibited by knocking down c-Jun or ATF2 expressions, induction of VEGF expression was partially attenuated in Beas-2B cells.Conclusion AP-1 is a critical transcriptional factor in mediating PM2.5-induced VEGF expression and inflammatory responses in human bronchial epithelial cells.

Clinical Efficacy of Folic Acid Intervene in Hyper-homocysteinemia Patients Combining Coronary Artery Disease and Heart Failure / 中国循环杂志

Objective: To observe clinical efficacy of oral folic acid (FA) intervene in hyper-homocysteinemia (HHcy) patients combining coronary artery disease (CAD) and heart failure (HF), to study the effect of blood level of Hcy on cardiac function. Methods: A total of 126 relevant patients with blood level of Hcy>15 μmol/L were randomly divided into 2 groups:Routine group, the patients received anti-platelet therapy, statins, beta-blockers, diuretics, angiotensin converting enzyme inhibitor (ACEI) or angiotensin II receptor antagonist and FA group, in addition to above mentioned therapies, the patients also received FA 5 mg/day. n=63 in each group and all patients were treated for 3 months. Fasting blood levels of Hcy, BNP and left ventricular end diastolic diameter (LVEDD), left ventricular ejection fraction (LVEF), 6-minute walk test (6MWT) were compared between 2 groups at pre- and 3 months post-treatment. Results: ① Based on NYHA classification, the patients with cardiac function at II, III, IV had accordingly increased blood levels of Hcy, BNP and LVEDD, while decreased LVEF and 6MWT, all P<0.05. ② Blood levels of Hcy were positively related to BNP (r=0.733, P<0.001) and LVEDD (r=0.511, P<0.001), negatively related to LVEF (r=-0.382, P<0.001) and 6MWT (r=-0.410, P<0.001). ③ With 3 months treatment, FA group and Routine group showed decreased Hcy level as (8.43 ± 1.87) μmol/L vs (3.29 ±1.68) μmol/L and BNP (891.84 ± 456.10) pg/ml vs (682.24 ± 463.79) pg/ml, reduced LVEDD (4.33 ± 1.231) mm vs (2.06 ± 1.73) mm, while elevated LVEF (6.59 ± 2.28) % vs (2.52 ± 2.37) % and 6MWT (142.97 ± 55.15) m vs (86.35 ± 59.06) m, all P<0.05. Conclusion: Increased blood level of Hcy is risky for HF occurrence, FA may treat HHcy and further improve the cardiac structure and function in HF patients.

Characterization of a Salmonella typhimurium SL1344 cya mutant strain / 中国免疫学杂志

Objective:To explore the function of the cya gene and the preliminary mechanism of attenuated strain.Methods:The biological characteristics of cya mutant in acid and alkali resistant,salt resistance,motility,biofilm components,poisonous to the cells of epithelial cell adhesion,invasion were analysis.Results:The mobility capabilities,acid and alkali resistance and salt tolerance of cya mutant were significantly lower than the parent strain;the composition testing revealed that the cya mutant did not produce cellulose,curli and biofilm;at the same time the adhesion and invasion to epithelial cells of cya mutant had a prominent depression,and the toxicity to HeLa cells was weaker than the parent strain.Conclusion:The function of cya gene is closely related to athletic ability, penetration of cell membrane, the formation biofilm and virulence.It will provide a theory reference to the functional research of Salmonella typhimurium cya gene and the mechanism of attenuated strain.This will contribute to the development of oral vaccine using attenuated Salmonella typhimurium as vector.

Cardiovascular responses in elderly hypertensive patients with discopotracheal intubation / 实用医学杂志

Objective To compare cardiovascular responses of intubations by Discopo and Macintosh laryngoscope in elderly patients with hypertension. Methods Sixty elderly hypertensive patients with ASAⅡ~Ⅲundergoing elective surgery were equally randomized into two groups. After induction of general anesthesia , orotracheal intubation was performed with Discopo or Macintosh. The Mean arterial pressure (MBP) and heart rate (HR), as while as Electrocardiogram (ECG) and Blood oxygen saturation (SpO2), were recorded before (baseline values) and at 1,3,5 minutes after intubation(post-induction values). The rate of one-time successive intubation,intubation time and complication were also recorded. Results Compared with the macintosh group, the rate of one-time successive intubation was higher and the intubation time was shorter in the Discopo group (P< 0.05, respectively),with no complications. Compared with T0,MBP and HR at T2 in the two groups and T3 in the Macintosh group increased significantly (P < 0.05). Compared with the Macintosh group,MBP and HR at T2,T3 of the Discopo group decreased significantly (P < 0.05). Conclusion Little effect of tracheal intubation with Discopo on cardiovascular response was observed in elderly hypertensive patients.