Chitosan(CS)surface was modified with hydrophobic octyl groups to prepare N-octyl chitosan(nitro-gen-octyl chitosan;OC).Then hydrophilic group carboxyl-polyethylene glycol-amino (PEG);tumor-targeting lig-and D-glucosamine(DG);and membrane-penetrating peptide 9-D-arginine(9R)were linked to OC successively. Then the DG and 9R modified chitosan micelle (DG/9R-PEG-OC)with tumor-targeting and transmembrane effect was prepared.By hydrogen nuclear magnetic resonance spectrometer (1 H NMR)and sodium dodecyl sul-fate polyacryl amide gel electrophoresis(SDS-PAGE);the successful formation of DG/9R-PEG-OC was certified;with particle size of 151.8 nm and Zeta potential of 16.5 mV.The morphology of chitosan micelle observed by transmission electron microscope was homogeneous spherical structure.The drug loading content (DLC)(using fluorescein as a model drug)and encapsulation efficiency (EE)were about 28.2% and 75.0% measured by UV-visible spectrophotometer.Meanwhile;the drug showed a controlled releasing profile out of the micelle.Cellu-lar uptake experiments indicated DG/9 R-PEG-OC micelle had a significant tumor-tageting and transmembrane effects;especially on HepG2 cells;which exbihited high expression of the glucose transporter.Thus DG/9R-PEG-OC micelle could be a promising drug targeted delivery system of hydrophobic antitumor drugs.

[Objective]: To study the in-vitro anti-HBV activity of Bushen Jianpi Formula (BJF). [Methods] 2.2.15 cell strain was cultured in vitro to observe the effect of BJF on its secretion of HBsAg and HBeAg. [Results] After the addition of BJF for ten days, median toxic concentration of BJF was 37.80g/L, median inhibitory concentrations of HBsAg and HBeAg were 19.05g/L and 9.55g/L, and the therapeutic index was 1.98and 3.96. [Conclusion] It is suggested that BJF has an in-vitro inhibitory effect on the secretion of HBeAg.

We prepared gold nanostar (GNS) through seed growth method firstly,then formation of COX-2 siRNA(siCOX-2) and GNS composite modified with polyethylene glyco (PEG),2-amino-2-deoxy-D-glucos (DG) and 9-D-arginin (9R) was prepared.Mterwords,paclitaxel temperature sensitive liposome (PTX-TSL) was prepared by film dispersion method.Finally,siCOX-2 delivery systerm (PTX-TSL-(siCOX-2(9R/DG-GNS)))was obtained by hydrosulfuryl ligand reaction between siCOX-2 (9R/DG-GNS) and PTX-TSL The successful build of siCOX-2 (9R/DG-GNS) was vetified by nuclear magnetic resonance (NMR),sodium dodecyl sulfate polyacryl amide gel electrophoresis (SDS-PAGE),and ultraviolet spectrophotometry and agarose gel electrophoresis method.Particle size of PTX-TSL-(siCOX-2(9R/DG-GNS)) was (292 ± 14) nm and Zeta potential was about -(2.59 ± 0.12) mV,which were measured by Zetasizer Nano ZS90.The morphology of PTX-TSL-(siCOX-2 (9R/DG-GNS)) measured by transmissionelectronmicroscopy showed homogeneous star structure with phospholipid bilayer on the surface,and it showed good thermal conversion efficiency under radiation of 808 nm laser.Differential scanning calorimetry test showed that PTX-TSL phase transition temperature is about 42.6 ℃.The drug loading content(using dialysis method) and encapsulation efficiency of PTX-TSL were about 7.5％ and 95.4％,at the same time,the release process experiment of PTX-TSL showed that it had a good temperature sensitive release performance.It is hopeful that this siCOX-2 system can be used for reducing drug resistance of PTX and improving the treatment effect of PTX through the synergistic antitumor drug resistance effect of siCOX-2.

Objective To preliminarily investigate the protective effect of chronic clonorchis sinesis(Cs) infestation against sepsis in Sprague Dawley(SD) rats in order to explore its underlying mechanism.Methods Chronic Cs infestation model of SD rats was reproduced by intra-gastric administration with Cs ova.Twenty rats were randomly(random number) divided into normal group(n=10) and Cs group(n=10).The proportion of differentiation in M1 and M2 macrophages were detected by flow cytometry.The expressions of Arg-1(arginine-1),FIZZ 1,iNOs and TNF-αmRNA were examined by reverse transcriptase polymerase chain reaction(RT-PCR).The cecal ligation and puncture(CLP) procedure was performed to reproduce sepsis model of SD rats.Sixty rats were randomly(random number) divided into control group,SHAM group,CLP group,Mφ+CLP group,Cs-Mφ+CLP group,and Cs-CLP group.The cumulative mortalities were calculated.The pathological changes of the lung tissue in different groups were demonstrated by HE staining.The serum levels of cytokines TNF-α and IL-10 were detected by ELISA at 0,24,48 and 72 h after CLP procedure.Results Compared with M1 peritoneal macrophages differentiation in control group(91.9%),rat peritoneal macrophages were activated to M2 differentiation(95.1%) in chronic Cs infection group.RT-PCR assay showed expression of Arg-1 and FIZZ 1 mRNA were higher in M2 macrophages,and on the contrary, the expression of iNOS mRNA expression was higher in M1 macrophages.The expression of TNF-α mRNA in M1 was significantly higher than that in M2, whereas the expression of IL-10 mRNA in M2 was higher than that in M1.The cumulative mortality of septic rats 72 h after CLP procedure were much lower in both chronic Cs infestation group and M2 macrophages adoptive transfer group(CLP group 70%vs.Mφ+CLP group 50%vs.Cs-Mφ+CLP group 30%vs.Cs-CLP group 0%,P<0.05).In these two groups,the pathological damages in lung tissues were significantly improved.The serum level of TNF-α was decreased and the anti-inflammatory IL-10 level was increased significantly in these two groups with Cs compared with other groups.Conclusion M2 macrophages polarization induced by chronic Cs infestation with M2 phenotype gene and expression of anti-inflammatory cytokine gene play key role in increasing anti-inflammatory cytokines and decreasing pro-inflammatory cytokines to allerviate organ damage and ameliorating the survival rate in septic rats.

We transfected human neural stem cells using lentiviral vectors encoding glial cell line derived neurotrophic factor (GDNF) to study its expression level in vitro and to get a stable cell line expressing GDNF. First, GDNF gene was sub-cloned into the lentiviral transfer vectors. Then, the recombinant lentiviral supernatants were packaged by 293T cells through three plasmids transient co-transfection method using standard lipofectamine reagent. The viral titers were tested by the transfection efficiency of 293T cells. At the same time, human neural stem cells (hNSC) were transfected under different multiplicity of infection. GDNF gene expression level and protein secretion level of hNSC were tested by real-time PCR and ELISA methods after transfection. Lentiviral vectors encoding GDNF were constructed. Using lentiviral vectors encoding GDNF we successfully transfected human neural stem cells, and got a stable neural stem cell lines over-expressing GDNF. Furthermore, the results indicated that GDNF expression was influenced by the multiplicity of infection. Human neural stem cells could over-express GDNF through lentivial vectors tranfection. Its gene expression level and protein expression level correlate with the multiplicity of infection.
Embryonic Stem Cells ; metabolism ; Genetic Vectors ; genetics ; Glial Cell Line-Derived Neurotrophic Factor ; biosynthesis ; genetics ; Humans ; Lentivirus ; genetics ; Nerve Tissue ; cytology ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection

Objective:To investigate the clinical features of patients with anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis misdiagnosed as mental disorder, improve the early diagnosis rate and reduce misdiagnosis.Methods:The clinical data of patients with anti-NMDA receptor encephalitis diagnosed at the First Affiliated Hospital of Zhengzhou University from 2012 to 2018 were collected. Patients misdiagnosed as mental disorders were screened out. Their psychiatric symptom characteristics, disease course characteristics, imaging and laboratory findings, treatment and prognosis were retrospectively analyzed.Results:A total of 121 cases of anti-NMDA receptor encephalitis were collected, and 43 cases of mental disorders were screened out. Sixteen of the 43 patients (37.2%) had prodromal symptoms, and all the patients had psychiatric behavioral abnormalities (100%), including 32 cases (74.4%) of seizures, 13 cases (30.2%) of decreased level of consciousness, 21 cases (48.8%) of involuntary movements, 15 cases (34.9%) of decreased memory, 8 cases (18.6%) of speech dysfunction, and 8 cases (18.6%) of other neurological symptoms (central hyperventilation, autonomic dysfunction). Memory loss was observed in 15 cases (34.9%), speech dysfunction in 8 cases (18.6%), other neurological symptoms (central hypoventilation, autonomic dysfunction) in 8 cases (18.6%), and various symptoms may appear simultaneously or successively in the same patient. Thirty-eight cases had complete resolution of symptoms or only minor physical impairment, and 5 cases had recurrent admissions with mental abnormalities and seizures. The recurrence rate accounted for 11.6% (5/43).Conclusions:The clinical manifestations of anti-NMDA receptor encephalitis are complex and varied. Most of them have mental behavior abnormalities as the first symptom, which is easily misdiagnosed as mental disorder and delayed treatment will lead to prolonged disease course and poor prognosis.