Objective To investigate the protective effect of allicin on intestinal mucosal barrier of septic rats so as to explore the possible mechanism.Methods Twenty-four male SD rats were randomly (random number) divided into sham,septic model and allicin treatment group.Septic model was established by cecal ligation and puncture (CLP) in rats.Rats in the treatment group were administered with allicin (30 mg/kg,ip)at 6 h and 12 h after modeling,while those in the model and sham groups were treated with equal amount of saline instead.Rats were sacrificed at 24 h and the serum D-lactic acid,diamine oxidase (DAO) and fluorescence isothiocyanate-dextran (FITC-Dextran,FD-40) were determined to evaluate the intestinal mucosal barrier function.The levels of tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6),malondialdehyde (MDA),and the activity of superoxide dismutase (SOD) in intestinal tissue were measured.Histopathological changes of intestinal mucosa injury were assessed by Hematoxylin-eosin staining.Results Compared with the sham group,levels of serum D-lactic acid,DAO and FD-40 increased significantly in the CLP group (D-lactic acid:599.4±101.1 vs.149.2±20.63 nmoL/mL,t=11.84,P＜0.01;DAO:302.1 ±64.5 vs.76.57±14.76 ng/mL,t=9.433,P＜0.01;FD-40:6664.0±1437.0vs.1446.0±205.0 ng/mL,t =9.704,P ＜0.01);intestinal morphology damage occurred in the CLP group;intestinal levels of TNF-α,IL-6 and MDA increased greatly (TNF-αt:186.35 ±20.43 vs.58.76 ±8.94 pg/mL,t=17.23,P＜0.01;IL-6:763.25±85.23vs.125.36±14.37 pg/mL,t=22.54,P＜0.01;MDA:29.36±3.27vs.7.24±0.85 nmol/mg prot,t=16.61,P＜0.01),while SOD activity reduced (35.75±6.53 vs.73.26 ±8.35 U/rmg prot,t =10.57,P ＜0.01) in the CLP group.Allicin treatment greatly inhibited the increase of D-lactic acid,DAO and FD-40 levels in rat plasma caused by CLP (D-lactic acid:330.1 ±81.77 vs.599.4±101.1 nmol/mL,t=7.086,P＜0.01;DAO:171.8±49.70vs.302.1±64.56ng/mL,t=5.45,P＜0.01;FD-40:3349.0±1167.0 vs.6664.0±1437.0 ng/mL,t=6.165,P＜0.01);intestinal morphology damage was improved in the allicin treatment group;allicin treatment greatly inhibited the intestinal levels of TNF-o,IL-6 and MDA and preserved the intestinal SOD activity compared with the CLP group (TNF-α:95.37 ±12.68 vs.186.35 ±20.43 pg/mL,t =12.29,P＜0.01;IL-6:354.27±46.27vs.763.25±85.23pg/mL,t=14.45,P＜0.01;MDA:16.27±3.14vs.29.36±3.27 nmol/mgprot,t=9.831,P＜0.01;SOD:55.35 ±6.23vs.35.75±6.53 U/mgprot,t=5.522,P ＜0.01).Conclusions Allicin could inhibit local inflammation and oxidative stress in the intestine and exerts protective effect on intestinal mucosal barrier of septic rats.

Objective To construct the lentivirus carrying human ?-catenin-EGFP(enhanced green fluorescent protein)and observe its expression in human follicle stem cells.Methods The ?-catenin gene sequence was amplified by RT-PCR from extraction of total RNA of human vascular endothelial cells.TA cloning technique was utilized to acquire gene subcloned pUCm-T-?-catenin.After transformation reaction,candidate clone was further analyzed by PCR and gene sequencing.Then the plasmid was transfected into FT293 cells.After identification by Western blotting,the plasmid was transfected into FT293 cells again for packaging.Infection titer was monitored by green EGFP expression.The expression of ?-catenin-lentivirus in human follicle stem cells were observed under inverted fluorescence microscope.Results The ?-catenin gene was cloned into the lentivirus successfully.The high expression of green fluorescence protein in FT293 cell line was found under fluorescent microscope.Viral titer checked by real-time PCR was about 2.0?108 TU/mL.When the multiplicity of infection(MOI)was 10,the infection efficiency of ?-catenin-lentivirus in human follicle stem cells was nearly 80% after infection 48 h around.After 3 weeks of continuous observation,we found the infection efficiency still keeping in the range of 80%-90%.Conclusion The lentivirus expression vector for ?-catenin was successfully constructed.It can steadily infect human follicle stem cells and the infection efficiency is considerable high.

Background Optic neuritis is closely associated with multiple sclerosis (MS).Its pathogenesis is uncompletely clear,and less basic researches are carried out at home and abroad. Objective This study was to reveal the expression of myelin basic protein (MBP) in the optic nerve of rat with experimental allergic encephalomyelitis (EAE) and to provide a theoretical evidence for the research of the relationship of optic neuritis with MS. Methods Fifty clean Wistar rats were randomized into the control group and immune 8,12,18 and 25 days groups.Myelencephalon was collected from 5 guinea pigs to prepare the homogenate and mixed with the isovolumetric complete Freud' s adjuvant (CFA).The 0.5 ml mixed antigen emulsifier was subcutancously injected into the 4 maps together with Bordetella pertussis 0.2 ml under the cutancous of dorsalis pedis at 0 and 48 hours to induce the EAE.Behavior of the rats was evaluated to score the neurological function.The optical nerve sections were prepared 8,12,18 and 25 days after immunology for the histopathological examination,and immunochemistry and Western blot were used to detect the expression of MBP in optic nerve.The use of the animals complied with the Regulation for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission. Results The disorder of motor nerve was seen 12 days following the immune,and the clinical neural functional scores were significantly higher 12 day and peaked on 18 days myelination and then gradually reduced.The histopathological examination showed that the irregular alignment of neural fiber was found at 12 days,and changes of cellular structure,edema of neural shaft bunch were observed at 18 days.However,the abnormal cells were significantly less 25 days following the immune.Immunochemistry showed that the MBP was expressed mainly in the myelination of optic nerve fibers.The numbers of positive cells for MBP were (115.75±26.49)cells/5 fields at the 12th day,showing a significant lowing in comparison with (167.44±22.49)cells/5 fields of control group (t=4.537,P＜0.05 ).The positive cells were lest at the 18th day with the values ( 75.57 ± 34.54) cells/5 fields ( t =6.362,P＜0.01 ).At the 25th day,positive cells increased to ( 117.63 ± 13.78) cells/5 fields,which still was lower than those of the control group ( t =4.068,P＜0.05 ).Western blot assay illuminated that with the prolong of immune time,MBP/β-actin ratio in optic nerve was gradually reduced and followed the same pattern at the 12th day( t =4.639,P＜0.05 ).At 18 days after immune in comparison with the control group,the expression of MBP/β-actin ratio in optic nerve was the least (t=8.427,P＜0.01). Conclusions MBP can be degraded in rat optic nerve.This is a further evidence that optic neuritis is a severe demyelination disease.It is clearly related to MS.

Cell Proliferation ; DNA ; chemistry ; Durapatite ; chemistry ; Hep G2 Cells ; Humans ; Liposomes ; chemistry ; Nanoparticles ; chemistry ; Particle Size ; Recombinant Proteins ; chemistry ; Transfection

Objective To study the relationship between arterial compliance and diastolic dysfunction in essential hypertensive patients.Methods Fifty-one treated hypertensive patients and 47 normotensive controls were enrolled.Arterial stiffness was assessed by vascular e-TRACKING technology of ultrosonic.Tissue Doppler echocardiography combined with conventional Doppler echocardiography was used for assessment of left ventricular diastolic function and peak late diastolic velocity of mitral flow.The ratio of E/A,Em/Am and E/Em were measured.Results Compared with normotensive controls,hypertensive patients have increased arterial sclerosis index(?) [(10.5?3.8) vs control(8.1?3.4),P

Objective To approach the neurobiochemical mechanism of chronic central pain (CCP) after spinal cord injury (SCI). Methods 28 SD rats were divided into four groups, the normal group (group A), the pseudosurgery group (group B), and groups with CCP (group C) and without CCP (group D) after L1 spinal cord section injured with WADE method. T13 and L2 segments of rats' spinal cord were took and concentration changes of substance P (SP) in the spinal dorsal horn between two sections were examined by immunofluorescence histochemistry staining combined with confocal laser scanning microscope. Results Concentration of SP in the group D was decreased significantly compared with groups C,A and B (P<0.05-0.01), that of the group C was less than that of group A and B (P<0.05). Conclusion The rat model established by WADE method is proper to study CCP after SCI. SP in dorsal horn of spinal cord may inhibit the CCP after SCI in some degrees.

Objective To investigate the correlation between left ventricular remodeling and systolic function by mid-wall fractional shortening(mFS).Methods 51 cases of hypertension and 47 cases of healthy volunteers were enrolled and systolic parameters were measured as ejection fraction(EF), fractional shortening(FS) and mFS. Acorrding to left ventricular remodeling parameteres inclulding LVMI and RWT, hypertension group was divided into four subgroups. The correlation among these parameters were analyzed.Results Thicker left ventricular wall and lower mFS in patient with hypertension compaired with those in controll (P<0.05). There were negative correlation between mFS and RWTr=-0.42, P<0.05) and positive correlation between mFS and LVMI r=-0.67, P<0.01).Conclusion mFS is a valuable parameter compared with traditional parameteres such as EF and FS. There is a linear association between parameteres of ventricular remodeling and systolic function assessed by mFS in early stage of hypertension.

Using of clearing zones on pachyman agar medium, there are 217 plant endophytic actinomycetes, producing ?-1,3-glucanase were screened. 45.6% of the strains produced ?-1,3-glucanase, in which the strains from cucumber are up to 38. The percentages of endophytic actinomycetes from different hosts produceing ?-1,3-glucanases were different. The percentage of the strains in Rhizoma Polygonatum produced ?-1,3-glucanases is the highest, up to 88.9%. The Inhibited effects of plant endophytic actinomycetes which produced extracellular ?-1,3-glucanases on mycelium growth of Sclerotinia sclerotiorum were detected in vitro. Cucumber endophytic actinomycete gCLA4 strain was screened out from 99 isolates, which can strongly inhibit the growth of S. sclerotiorum. The optimal ?-1,3-glucanases fermentation conditions of strain gCLA4 were investigated, they were pachyman 0.2%, peptone as nitrogen, pH 7～8 for 5 days. The ?-1,3-glucanases of strain gCLA4 had some inhibiting efficiency on 13 plant pathogens, in which inhibiting efficiency to Botryosphaeria dothidea was the strongest.

Methods: A total of 10 specific pathogen free (SPF) grade female DBA/2J mice were randomly divided into model group and QGA II group (n = 5 for each group), while additional 5 SPF-grade female C57BL/6J mice were assigned to control group. Mice presented spontaneous high IOP and showed elevated approximately at the age of seven months. The high IOP was maintained until week 38, when gavage was initiated. Mice in control group underwent the same intragastric treatment, while those in QGA II group were gavaged with QGA II (9.67 g/kg), once a day for four weeks. Retinal morphology was examined using hematoxylin and eosin (HE) staining, with the number of retinal ganglion cells (RGCs) counted. The expression level of Brn3a protein, a specific marker for RGCs, was detected by immunofluorescence, with the mean optical density (OD) measured for quantitative analysis. In addition, 16S rDNA sequencing was leveraged to analyze changes in the diversity of gut microbiota, including their α-diversity (Chao1, Shannon, Pielou’s evenness, and observed species index) and β-diversity. Venn diagrams and linear discriminant analysis effect size (LEfSe) analysis was employed to investigate the number of amplicon sequence variants (ASVs), the abundance of differential gut microbiota species, and the classification of species at both the phylum and genus levels within the three groups of mice. Results: HE staining revealed that compared with control group, model group showed significant reduction in the number of RGCs (P < 0.01), with intracellular vacuolar degeneration and nuclear pyknosis. After QGA II treatment, the number of RGCs was significantly increased compared with model group (P < 0.01), with notable improvements in intracellular vacuolar degeneration. Immunofluorescence analysis showed that the mean OD of Brn3a protein was significantly decreased in model group compared with control group (P < 0.01), while QGA II treatment significantly elevated its expression level (P < 0.01). Analysis of α-diversity showed that after QGA II intervention, the Chao1, Shannon, and Pielou’s evenness indices were significantly increased (P < 0.01), and the observed species index was elevated (P < 0.05). β-Diversity analysis demonstrated distinct clustering among the three groups, indicating relatively low similarity in bacterial community structures. ASV clustering identified a total of 14 061 ASVs across all groups, with 9 514 ASVs shared between model and QGA II groups. At the phylum level, the abundance of Bacteroidetes was significantly decreased in model group compared with control group (P < 0.01), while Firmicutes and the Firmicutes/Bacteroidetes (F/B) ratio were significantly increased (P < 0.01). QGA II treatment significantly reduced both Firmicutes abundance and the F/B ratio (P < 0.01). At the genus level, Lactobacillus was dominant across all groups, with its abundance significantly increased in model group (P < 0.01) and subsequently decreased following QGA II intervention (P < 0.05). Conclusion QGA II restructured the gut microbiota of DBA/2J mice with chronic high IOP, bringing changes in their diversity and abundance of components. Firmicutes, Bacteroidetes, Lactobacillus, along with their associated microorganisms, are likely critical components of the gut microbiota that contribute to the optic neuroprotective effects of QGA II on chronic high IOP mice.

To study the inhibitory effect of RNA interference (RNAi) on MMP-9 gene expression in THP-1 cell line.To investigate the application of RNAi on the therapy of leukemia.Methods:Small interfering RNA ( siRNA) for MMP-9 gene was designed and transfected into THP-1 cells.MMP-9 mRNA expression was assessed by RT-PCR, and MMP-9 protein expression was tested by Western blot.MTT and trypan blue staining were used to observe the effect on the proliferation of THP-1 cells after RNAi.The changes in cell morphology were observed under the microscope.Results:The expressions of MMP-9 mRNA and protein were inhibited in THP-1 MMP-9 siRNA-transfected cells ,significantly lower than those of control cells.The results of MTT and trypan blue staining in-dicated that the proliferation ability of THP-1 cells obviously decreased after siRNA-transfected 48h and 72h.The growth of cells was in-hibited and the cells survival rate was significantly lower than that of control group ( P<0.05 ).The cells of control groups grew semi-quote wall under inverted microscope.The outline of cells was clear and the shape was uniform.The cells grew vigorously.While the growth of cells in siRNA group was inhibited.The morphology of siRNA group cells changed obviously by the Wright staining.Most cells expressed changes of apoptosis.Conclusion: siRNA for MMP-9 gene can not only reduce the expressions of MMP-9 mRNA and protein,but also inhibit the proliferation and induce apoptosis of THP-1 cells.