ObjectiveToinvestigatethecharacteristicsofsinglelacunarinfarct(SLI)andipsilateral multiple lacunar infarction (MLI), and the differences of risk factors and and pathologies betw een them. Methods The clinical data of al patients w ith cerebral infarction in acute internal carotid artery territory from August 1, 2008 to December 13, 2014 w ere analyzed retrospectively. Lacunar infarctions w ere screened according to the clinical manifestations and imaging findings. The patients w ere divided into a SLI, a unilateral MLI in the same blood supply area (MLI 1) and a unilateral MLI in the different blood supply area (MLI 2) group according to the number and location of the lesions show ed on diffusion w eighted imaging. Multivariate logistic regression analysis w as used to identify potential independent risk factors. Results The incidences of ipsilateral carotid plaque (73.33%vs.48.67%; χ2 =5.801, P=0.016), ipsilateral unstable carotid plaque ( 70.0%vs.42.5%; χ2 =7.192, P= 0.007 ), and ipsilateral carotid stenosis ≥50%(16.67%vs.1.77%; χ2 =8.327, P=0.004) of the MLI 1 group w ere significantly higher than those of the SLI group; the incidence of atrial fibril ation of the MLI 2 group w as significantly higher than that of the SLI group (40.0%vs.0.88%; χ2=15.887, P<0.001); there w ere no significant differences in the remaining risk factors among each group. Multivariate logistic regression analysis showed that atrial fibrilation (odds ratio [OR] 14.452, 95% confidence interval [CI] 1.558-134.011; P=0.019) and ipsilateral carotid stenosis ≥50% (OR 11.483, 95%CI 2.202-59.891; P=0.011) w ere the independent risk factors for MLI. Conclusions MLI may have different risk factors and pathogeneses w ith SLI. Atherosclerotic lesions and embolism are the important pathogeneses of MLI, w hile SLI is not.

A real-time polymerase chain reaction ( PCR ) micro total analysis system (μ-TAS ) integrated nucleic acid extraction, PCR amplification and real-time-fluorescent PCR detection on a same microfluidic chip was prepared for the fully automated and on-chip analysis of nucleic acid. The proposed method had the advantage such as low sample consumption, fast analysis and simple operation and so on. Micromachining technology was used to fabricate the anodic molds of integrated nucleic acid extraction microfluidic chip. A polydimethylsiloxane (PDMS) substrate with 3D channels was manufactured by a combination of molds and an injection molding method. The glass substrate and the chip were bonded together using a plasma treatment. The μ-TAS included a microfluidic control device whose micro fluidic velocity ( 0-10 mL/min ) could be adjusted, a TEC platform which the precision of temperature control was 0. 1℃ and a CCD detection module. The DNA of human blood was extracted by using a silica gel membrane method on the microfluidic chip. The processes of DNA extraction and detection were preset in the μ-TAS. Human blood lysate ( 20 μL ) were driven to the extraction chamber and was then washed. The fluidic drive speed was 2 mL/min. DNA and PCR reagents were mixed and then were driven into the PCR chamber. The fluidic drive speed was 1 mL/min. The GAPDH gene in extracted genome DNA was amplified by PCR and detected. The amplified product was verified by melting analysis. The results of nucleic acid extraction method on the chip were compared to those obtained using a standard manual centrifuge extraction method. The amplification curves were obvious. Ct values of the chip method were 25 . 3 and 26 . 9 . The denaturation temperature of all the melting was 89 . 9 ℃. The results validated that the chip-based method and device realized the extraction of nucleic acid, amplification and detection automatically.

This study reported a 50-year-old female patient who was diagnosed with anti-IgLON family member 5(anti-IgLON5)antibody-related encephalopathy,presented with cognitive and sleep disorders,autonomic dysfunction and seizures,positive serum IgLON5 antibody but negative cerebro-spinal fluid IgLON5 antibody,negative human leukocyte antigen(HLA)by genetic testing,and was diagnosed as anti-IgLON5 antibody-related encephalopathy.After hospital admission,the patient was given intravenous methylprednisolone combined with immunoglobulin immunotherapy,donepezil for improvement of cognition,sodium valproate and oxcarbazepine for prevention and treatment of epilep-tic seizures,and finally her symptoms improved significantly.

This study reported a 50-year-old female patient who was diagnosed with anti-IgLON family member 5(anti-IgLON5)antibody-related encephalopathy,presented with cognitive and sleep disorders,autonomic dysfunction and seizures,positive serum IgLON5 antibody but negative cerebro-spinal fluid IgLON5 antibody,negative human leukocyte antigen(HLA)by genetic testing,and was diagnosed as anti-IgLON5 antibody-related encephalopathy.After hospital admission,the patient was given intravenous methylprednisolone combined with immunoglobulin immunotherapy,donepezil for improvement of cognition,sodium valproate and oxcarbazepine for prevention and treatment of epilep-tic seizures,and finally her symptoms improved significantly.

Objective To prepare SDF-1 α-loaded nanoliposome ( SNP )-SonoVue complex and investigate its tracing abilities, sustained-release property and effect on migration of bone marrow mesenchymal stem cells (BMSCs). Methods The SNP was prepared to detect its physical characteristics including particle size,zeta potential, morphology, encapsulation efficiency and drug loading.SNP-SonoVue was constructed to detect the sustained release situation of SNP and SNP-SonoVue after low frequency ultrasound ( LIFU ) irradiation, and the connection of SNP-SonoVue was observed by fluorescence microscope. Effects of SNP-SonoVue on migration of BMSCs were detected to evaluate its bioactivity. BMSCs were divided into 6 groups,including Group A: SDF-1α+ 1% serum medium;Group B: SNP- SonoVue+ 1% serum culture medium;Group C:SNP-SonoVue+ 1% serum culture medium + LIFU ( 1 MHz,0.5 W/cm2, expose 30 s stop 30 s, 4 min);Group D: BNP-SonoVue+1% serum medium;Group E:BNP-SonoVue+1% serum medium+LIFU ( 1 MHz, 0.5 W/cm2, expose 30 s stop 30 s, 4 min),Group F:PBS+1% serum culture medium (control group). Its tracing abilitie were investigated in vitro. Results The average particle size of SNP was(220.4±9.9)nm,and the particle dispersion index(PDI) was(0.172± 0.015), the average zeta potential was ( 35.6 ± 1.7) mv. It was showed spherical dispersion by transmission electron microscopy. The encapsulation efficiency was up to 96.7% and the drug entrapment content was 481.76 ng/mg. Flow cytometric showed the suitable conditions for SNP-SonoVue preparation was that the ratio of SNP quality(mg) to Sono Vue microbubbles number(a) was20:(2.8×109)to40:(2.8× 109). Fluorescence microscopy showed that shells of SonoVue microbubbles connected with large numbers of SNP labeled with red fluorescent DiI. Drug release experiment showed that the cumulative SDF-1α release amount of SNP and SNP-SonoVue exposed to LIFU respectively were ( 68.61 ± 3.97 )% and ( 63.21 ± 5.68)% in vitro within 7 days, and the difference was not statistically significant ( P ＞ 0.05 ). Cell migration experiments confirmed that the transfer function of BMSCs in Group A, Ggroup B and group C was significantly higher than that in control group ( P ＜ 0.05 ), but there was no significant difference among the Group A, Ggroup B and group C ( P ＞0.05). In vitro development experiment showed that the SNP-SonoVue complex had obviously enhanced development effect. Conclusions SNP-SonoVue complex is successfully prepared. It has obviously enhanced development effect and can lead to migration of BMSCs.

Objective To observe the clinical manifestations of 3 cases with myelin oligodendro-cyte glycoprotein(MOG)antibody-associated disease and anti-N-methyl-D-aspartate receptor(NMDAR)encephalitis antibody overlapping syndrome(MNOS),aiming to expand the understanding of the clinical spectrum of such syndromes.Methods Retrospective analysis was performed on the data of 3 patients with MNOS who were positive for both MOG antibodies and NMDAR antibodies.Clinical features,neuroimaging characteristics,and outcomes were collected,and cell-based assay(CBA)tech-nique was used for diagnosis.Results One case presented both positive MOG antibodies and NMDAR antibodies,but the clinical manifestations were typical symptoms of anti-NMDAR encephalitis.In one case,the clinical and cranial magnetic resonance imaging(MRI)features of demyelinating disease re-curred after anti-NMDAR encephalitis,with atypical symptoms of MNOS such as numbness and weakness in limbs,blurred vision,and diplopia.The last case presented both positive MOG antibodies and NMDAR antibodies,but the clinical manifestations were typical symptoms of anti-NMDAR encephalitis.In MNOS,MOG antibody-associated disease and anti-NMDAR encephalitis may appear simultaneously or sequentially,with epilepsy being the most common symptom.Cranial MRI findings showed that the pa-tients presented and mainly involved supratentorial lesions,which may also involve the brainstem,but no spinal cord lesions were found.All patients showed slightly abnormal cerebrospinal fluid.Patients showed a good response to first-line immunotherapy during the acute phase of the disease,with a fa-vorable prognosis.But most patients were prone to relapse.Conclusion In MNOS patients,anti-NMDAR encephalitis may present with clinical and(or)MRI features of demyelinating disease simul-taneously or sequentially.The clinical manifestations of patients are complex and diverse.Patients with atypical symptoms require to improving the understanding of MNOS and timely treatment.

Objective To observe the clinical manifestations of 3 cases with myelin oligodendro-cyte glycoprotein(MOG)antibody-associated disease and anti-N-methyl-D-aspartate receptor(NMDAR)encephalitis antibody overlapping syndrome(MNOS),aiming to expand the understanding of the clinical spectrum of such syndromes.Methods Retrospective analysis was performed on the data of 3 patients with MNOS who were positive for both MOG antibodies and NMDAR antibodies.Clinical features,neuroimaging characteristics,and outcomes were collected,and cell-based assay(CBA)tech-nique was used for diagnosis.Results One case presented both positive MOG antibodies and NMDAR antibodies,but the clinical manifestations were typical symptoms of anti-NMDAR encephalitis.In one case,the clinical and cranial magnetic resonance imaging(MRI)features of demyelinating disease re-curred after anti-NMDAR encephalitis,with atypical symptoms of MNOS such as numbness and weakness in limbs,blurred vision,and diplopia.The last case presented both positive MOG antibodies and NMDAR antibodies,but the clinical manifestations were typical symptoms of anti-NMDAR encephalitis.In MNOS,MOG antibody-associated disease and anti-NMDAR encephalitis may appear simultaneously or sequentially,with epilepsy being the most common symptom.Cranial MRI findings showed that the pa-tients presented and mainly involved supratentorial lesions,which may also involve the brainstem,but no spinal cord lesions were found.All patients showed slightly abnormal cerebrospinal fluid.Patients showed a good response to first-line immunotherapy during the acute phase of the disease,with a fa-vorable prognosis.But most patients were prone to relapse.Conclusion In MNOS patients,anti-NMDAR encephalitis may present with clinical and(or)MRI features of demyelinating disease simul-taneously or sequentially.The clinical manifestations of patients are complex and diverse.Patients with atypical symptoms require to improving the understanding of MNOS and timely treatment.

ObjectiveTo investigate the effect of Bufeitang on intestinal flora of rats with lung Qi-deficiency syndrome of chronic obstructive pulmonary disease（COPD）， and to explore the mechanism of traditional Chinese medicine in regulating intestinal flora and thus restoring the balance of lung-gut axis. MethodA total of 84 rats were randomly divided into 7 groups， including blank group， model group， fecal bacterial transplantation（FMT） group， dexamethasone group and low， medium and high dose groups of Bufeitang， 12 rats in each group. Except for the blank group， cigarette and sawdust fumigation combined with intratracheal instillation of lipopolysaccharide（LPS） were used to establish the COPD rat model with lung Qi-deficiency syndrome in all other groups. The low， medium and high dose groups of Bufeitang were intragastric administrated with Bufeitang（3.645， 7.29， 14.58 g·kg^-1）， the FMT group was given fecal bacteria liquid enema（10 mL·kg^-1）， dexamethasone group was given dexamethasone acetate tablet suspension by gavage（0.135 mg·kg^-1）， the blank group and model group were given equal amount of distilled water. Fresh feces were collected after 28 d of continuous intervention for 16S rRNA gene sequencing. Lung and colon tissues were stained with hematoxylin-eosin（HE） for pathomorphological observation， and enzyme-linked immunosorbent assay （ELISA） was performed to detect the contents of tumor necrosis factor-α（TNF-α） and interleukin-8（IL-8） in lung tissues. ResultCompared with the blank group， the model group showed severe abnormal lung tissue structure with alveolar atrophy and collapse accompanied by severe inflammatory cell infiltration. Compared with the model group， the extent of injury was significantly improved， and inflammatory cell infiltration was reduced with basically normal alveolar structure in the high dose group of Bufeitang. Compared with the blank group， the model group had severely abnormal colonic tissue structure， the epithelial cells in the mucosal layer were eroded and shed， the number of inflammatory cells increased， the submucosal layer was edematous and the gap was enlarged. Compared with the model group， the extent of damage was significantly improved in the medium and high dose groups of Bufeitang， the epithelial cells in the mucosal layer were neatly and closely arranged， with only a small amount of inflammatory cell infiltration and no significant degeneration. Compared with the blank group， the TNF-α and IL-8 levels of lung tissue in the model group were significantly increased（P<0.01）. Compared with the model group， the TNF-α and IL-8 levels of lung tissues in the low， medium and high dose groups of Bufeitang were significantly decreased（P<0.01）. Bufeitang significantly modulated the number of bacteria species as well as alpha and beta diversity of model rats， corrected the return of intestinal flora to normal abundance and diversity， and positively regulated 4 differential phyla（such as Firmicutes， Proteobacteria） and 13 differential genera（such as Turicibacter， Lactobacillus， Anaerobiospirillum， Intestinimonas） in COPD model rats with lung Qi-deficiency syndrome， and down-regulated 2 carbohydrate metabolic pathway functions， including the pentose phosphate pathway（non-oxidative branch） Ⅰ and the Calvin-Benson-Bassham cycle. ConclusionBufeitang can modulate the abundance and diversity of intestinal flora species， affect the function of metabolic pathways， repair the structure of lung and colon tissues， regulate the level of inflammatory factors， and thus improve COPD with lung Qi-deficiency syndrome. The mechanism may be related to its regulation of inflammation-related intestinal flora to restore the balance of lung-gut axis in COPD with lung Qi-deficiency syndrome.

As one of the hallmarks of cancer, metabolic reprogramming leads to cancer progression, and targeting glycolytic enzymes could be useful strategies for cancer therapy. By screening a small molecule library consisting of 1320 FDA-approved drugs, we found that penfluridol, an antipsychotic drug used to treat schizophrenia, could inhibit glycolysis and induce apoptosis in esophageal squamous cell carcinoma (ESCC). Gene profiling and Ingenuity Pathway Analysis suggested the important role of AMPK in action mechanism of penfluridol. By using drug affinity responsive target stability (DARTS) technology and proteomics, we identified phosphofructokinase, liver type (PFKL), a key enzyme in glycolysis, as a direct target of penfluridol. Penfluridol could not exhibit its anticancer property in PFKL-deficient cancer cells, illustrating that PFKL is essential for the bioactivity of penfluridol. High PFKL expression is correlated with advanced stages and poor survival of ESCC patients, and silencing of PFKL significantly suppressed tumor growth. Mechanistically, direct binding of penfluridol and PFKL inhibits glucose consumption, lactate and ATP production, leads to nuclear translocation of FOXO3a and subsequent transcriptional activation of BIM in an AMPK-dependent manner. Taken together, PFKL is a potential prognostic biomarker and therapeutic target in ESCC, and penfluridol may be a new therapeutic option for management of this lethal disease.