Objective To investigate the protective function of cytoglobin (CYGB) on neurodevelopment of the neonatal rats with hypoxic-ischemic brain damage (HIBD).Methods Healthy seven day old Sprague-Dawlay rats were randomly divided into sham operated group,HIBD group,HIBD+Hemin group and HIBD+zinc protoporphyrin (ZnPP) group.The rats of HIBD,HIBD+Heminand HIBD+ZnPPgroup were given normal saline (0.5 ml),Hemin (50 mg/kg) and ZnPP (50 mg/kg) intraperitoneally respectively,and 12 hours later the left carotid arteries of these rats were ligated and cut off,then hypoxic treated for 2 hours to establish the HIBD models.At 0,24,48 h after HIBD models were established,the expressions of CYGB in the cerebral cortex and hippocampus were observed by immunohistochemistry analysis.At 48 h after HIBD,histopathological changes of brain were observed after HE staining.At 72 h after HIBD,the water content of the brain was observed.At 28 days after HIBD,long term study memory outcome was assessed by Morris water maze.Analysis of variance and Bonferroni test were applied as statistical methods.Results (1)The expression of CYGB in brain(expressed by average gray value which negatively correlated with protein levels):At 0 h,the average gray value of CYGB in the cerebral cortex in HIBD+ Hemin and HIBD group were 166.7±5.1 and 207.1±5.1,which were lower than that in sham operated group (232.3±3.4); but in HIBD+ZnPP group,it was higher (234.9±4.5)(P＜0.05).The average gray value of CYGB was decreased with the extension of hypoxic-ischemic time.At 48 h,the average gray value of CYGB was the lowest in HIBD+Hemin group (126.0± 2.6),followed by HIBD group (150.9±4.5) and HIBD+ZnPP group (163.7±6.3),and the highest was in sham operated group (232.1±5.8)(all P＜0.01).(2) Histopathologic changes of the brain:At 48 h,typical cerebral infarction and hemorrhage were seen in HIBD,HIBD+Hemin and HIBD+ZnPP group,but those were less severe in HIBD + Hemin group than in HIBD + ZnPP group.(3) The water content of the brain:At 72 h,the water content of the left brain in HIBD and HIBD+ZnPP group was (86.5±0.4)％ and (87.3±0.3)％,which was higher than that in right brain [(85.6±0.2)％ and (85.9±0.2)％] (t 12.57 and 11.32,P＜0.01,respectively).(4)Function of the hippocampus:Morris water-maze showed that the longest average escape latency in HIBD+ZnPP group [(76.7±29.8) s],followed by HIBD group [(71.0±30.5) s],HIBD+ Hemin group [(46.7±34.0) s],and sham operated group [(38.3±30.3) s] (all P＜0.01).(5) Long-term histopathologic changes of the brain:At 34 d,brain atrophy rate was the highest in HIBD+ZnPP group [(34.07± 6.75) ％],and then in HIBD group [(29.73± 6.53) ％] and HIBD+ Hemin group [(18.33±4.52)％],which were all higher than that in the sham operated group [(1.55±1.32)％](all P＜0.01).HE staining showed that the hippocampal stratum pgramidale was getting thinner and a large number of neurons was lost in HIBD and HIBD+ ZnPP group,but only a small amout of neurons was lost in HIBD+ Hemin group.Conclusions Increased expression of CYGB in HIBD brain could mitigate the short term and long term pathological injury,and protect the long-term study memory function of the hippocampus.

Objective: To investigate the clinical effect of Naofuqing Capsules on senile dementia. Methods: 30 patients with senile dementia were treated with Naofuqing Capsules and the curative results were compared with that of the control group 30 cases. Results: The results showed that the integral titres of MMSE and HDS R of groups increased obviously after treatment( P 0.05). The total effective rates were 83.3% and 80.0%,respectively. No obvious difference was observed in clinical effect between two groups. Conclusion: Naofuqing Capsules possesses a definite therapeutic effect for senile dementia.

Objective To investigate the expression and change of cytoglobin(Cygb)gene on hypoxicischemic brain damage(HIBD).Methods Fifty SD rats of 7days old were divided into four study groups and one control group.The brain tissues were taken at 4hours,12hours,24hours and 48hours after the onset of HIBD.Cygb mRNA was determined by the reverse transcription PCR.One-way method of GraphPad Prism was used for statistics.Results The fragment length of PCR products was identical with experimental design.The expression level of Cygb gene increased at 4h after ischemia,and peaked at 24h.48h after HIBD,the Cygb gene level began to decrease.Conclusion The expression of Cygb in brain tissue increased rapidly after HIBD of newborn rats,suggesting that Cygb may have important function in the protection process of HIBD.

Objective To observe the change of intestinal microflora on the process of nonalcoholic steatohepatitis(NASH),and to explore the synbiotics therapeutic effect on NASH.Methods Rats were administrated with high fat diet to establish NASH model.In the process of NASH rats modeling,the level of triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), low density lipoprotein (LDL), fasting blood sugar (FBS) and fasting insulin (FINS) was dynamically tested by automatic biochemical analyzer.The change of main intestinal flora was detected by 16 S rRNA fluorescence quantitative polymerase chain reaction.NAFLD activity score was calculated.HE staining was used to observe the hepaticpathological changes and the TLR4 expression was detected by using enzyme-linked immunosorbent assay and immunohistochemical method.Until the 4th,8th,10th weekin the process of NASH modeling, 10 rats were feeded with synbiotics for 2 weeks, and all of above indicators were tested and observed.Results 1)With the extension of a high-fat diet feeding time, the degree of hepatocyte steatosis obviously increased.NAFLD score was significantly heightened(P<0.01).2)Number of independent activities of rats significantly increased, the serological level of TG, TC, LDL, FBS and FINS were lower significantly after intervention with synbiotics for 2 weeks(P<0.05).3)Synbiotics intervention for two weeks significantly increased the amount of bifidobacterium and lactobacillus and decrease the amount of enterococcus significantly(P<0.05).4)The expression of TLR4 was gradually increased in the process of NASH rats modeling(P<0.05),but decreased after 2 weeks of the synbiotics-intervention (P<0.05).Conclusions Intestinal microecology change is closely related to the development of NASH,therefor, synbiotics could improve the quality of life and biochemical indicators of NASH rats through adjusting intestinal microecology and the expression level of TLR4 protein might been involved.

AIM: To investigate the multidrug resistance(MDR),reversal activity of 2-[4-(2-pyridin-2-yl-vinyl) henyl]-4,5-bis-(4-N,N-diethylaminophenyl)-1(H)-imidazole(FG020318) in a retinoblastoma subline SO-Rb50/VCR,which is resistant to vincristine.METHODS: The procedure of stepwise increase in drug concentrations was used to obtain SO-Rb50/VCR,which was resistant to 200 ?g/L vincristine. The chemosensitivity of this drug resistant cell line with and without FG020318 or cyclosporine A(CSA) were detected by MTT assay and the function of p-glycoprotein(P-gp) was examined by rhodamine 123 accumulation detected with flow cytometry(FCM).RESULTS: FG020318(2.5 ?mol/L) significantly reduced IC50 and increased the rhodamine accumulation in a concentration-dependent manner. It was much stronger than the positive control CSA in reversal of MDR.CONCLUSION: A new tumor MDR modulator FG020318 partly reverses MDR in SO-Rb50/VCR.It may be a promising new drug to tackling MDR.

Objective To investigate the correlations between miRNA and mRNA ( the regulatory effects of miRNA) in a rat model of trinitro-benzene-sulfonic acid (TNBS)/ethanol induced ulcerative colitis ( UC) .Methods TNBS and ethanol were used to induce the development of UC in rats .After the modeling procedure and oral administration of normal saline ( NS) for 14 days, rats from the control and model groups were dissected to collect the samples of colonic mucosa .General and histological evaluations were performed to validate the modeling of UC .The expression of miRNA was profiled using miRNA microarray .The target miRNAs that were closely related to the pathogenesis of UC were selected out according to the results of mi -croarray and related literatures .RT-PCR was performed to verify the differentially expressed miRNAs .The mirWalk database was used to predict the target genes of miRNAs .In order to verify whether the predicted results were in accordance with the actual results , the microarray technology was used for mRNA expression profiling .The genes that showed interactions with those miRNAs were screened out .The David database was used for gene annotation .An interaction net between miRNA and mRNA was formed .Results General and histological manifestation of colon tissue samples from the model group were in accordance with the features of UC.Sixty-eight miRNAs were identified to be differentially expressed in rats from the model group and the control group (fold change>2, P<0.05, expression mean>7).Six candidate miRNAs were selected as hav-ing close relations to the pathogenesis of UC referring to reported literatures , the expression of which was checked and verified by real-time polymerase chain reaction (PCR).Compared with the control group, 4 miRNAs (miR-146a-5p, miR-146b-5p, miR-126a-3p and miR-21-5p) were up-regulated (P<0.01, P<0.05) and 2 miRNAs (miR-200b-3p and miR-145-5p) were down-regulated (P<0.01) in rats with TNBS/ethanol induced UC.Four mRNAs (IL-6, Ccl5, Mapk3 and Smad7) that interacted with the 6 miRNAs were identified based on the results of target gene prediction of the above 6 miRNAs and gene expression pro-filing.The David database was used to annotate the interactions for elucidating their significance in the path -ogenesis of UC .Conclusion A miRNA can regulate many signaling pathways and a signaling pathway can also be regulated by many miRNAs .Therefore , simply inhibiting certain pathways may not radically stop the process of inflammation .Studying the functions of miRNAs and elucidating the correlations between miRNA and mRNA might fundamentally inhibit the development of UC .

BACKGROUND: The preparation and antibacterial activity of metal ionic zirconium phosphates has been systemically investigated now, but the applications are limited owing to the discoloration or the low antibacterial activity. Here we prepared new antibacterial agents of quaternized zirconium phosphates by introducing quaternary ammonium salt bactericidal agent with high-effective, broad-spectrum and low-toxic into sodium zirconium phosphate through an ion-exchange method.OBJECTIVE: To explore the component structure and antibacterial activity of quaternized zirconium phosphates.DESIGN, TIME AND SETTING: An in vitro observational experiment was performed at Research Laboratory of Department of Chemistry, Jinan University from June to August 2009.MATERIALS: Quaternized zirconium phosphates were prepared by introducing dodecyl dimethyl benzyl ammonium chloride into sodium zirconium phosphate through an ion-exchange method.METHODS: The mol ratios of quaternary ammonium cations to cation exchange capacity of sodium zirconium phosphate in reaction solutions were 0.25: 1,0.5: 1, 1.0: 1, and 1.5 : 1, respectively, and four kinds of quaternized zirconium phosphates containing different contents of quaternary ammonium cations (QZrP-1, QZrP-2, QZrP-3, QZrP-4) were prepared through an ion-exchange method.MAIN OUTCOME MEASURES: The component structure and heat resistance of samples were measured by using an IR spectrometer, an elemental analyzer and a thermal analyzer, respectively. The minimum inhibitory concentrations (MICs) and minimal bactericidal concentrations (MICs) of the samples against Escherichia coli (E. co/i) and Staphylococci aureus (S. aureus) were estimated by a tube broth method.RESULTS: Quaternized zirconium phosphates were prepared, and the quaternary ammonium cation content increased with increasing the concentration of quaternary ammonium cations in reaction solution. The mass fraction of quaternary ammonium cations of QZrP-1, QZrP-2, QZrP-3, and QZrP-4 was 3.70%, 5.00%, 6.96%, and 10.01%, respectively. The onset temperatures of the decomposition for quaternary ammonium cations in quaternized zirconium phosphates were all higher than 345 °C, and they were preferable thermal stability. The antibacterial activity was higher when the quaternary ammonium cation content of quaternized zirconium phosphates increased. For quaternized zirconium phosphates QZrP-3 containing 6.96% mass fraction of quaternary ammonium cations, showed excellent antibacterial activity against E. coli and S. aureus.CONCLUSION: Quaternized zirconium phosphates QZrP-3 containing 6.96% mass fraction of quaternary ammonium cations,exhibited excellent thermal stability and antibacterial activity.

Objective To explore the clinical value of transgastric natural orifice transluminal endoscopic surgery(NOTES)in diagnosing unexplained ascites.Methods The clinical data in 12 cases of unexplained ascites diagnosed by adopting transgastric approach NOTES and performed abdominal exploration and peritoneal biopsy in our hospital from November 2015 to July 2016 were retrospectively analyzed.The operative risk and clinical application value were evaluated by statistically analyzing the postoperative complications occurrence and the diagnosis rate of disease.Results The definite diagnosis rate reached 100％ verified by pathology after abdominal exploration and peritoneal biopsy,in which 8 cases(66.7％)were tuberculous peritonitis,2 cases(16.7％)were liver cirrhosis,1 case(8.3％)was peritoneal mesothelioma,1 cases(8.3％)was peritoneal metastatic carcinoma;2 cases appeared abdominal pain after operation,including 1 case of neutrophil ratio increase,symptoms and persistent time of abnormal laboratory indexes did not exceed 24 h,the incidence rate was 8.3％;no complications of abdominal cavity infection,incision bleeding and puncture site fistula occurred.Conclusion The transgastric NOTES for conducting abdominal exploration and peritoneal biopsy in the diagnosis of unexplained ascites has the advantages of small trauma,less complications and rapid postoperative recovery,possesses an important clinical application value.

OBJECTIVETo study the lignans components in rat serum after oral administration of Fuzheng Huayu decoction (FZHY), and to investigate the active ingredients in vivo.

METHODA rapid, sensitive and selective method using liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MSn) was established. The serum samples were extracted with ethyl acetate (EtOAc)for three times. The chromatographic separation was achieved on a Waters Atlantis T3 column by gradient elution using methanol and water containing 5 mmol x L(-1) ammonium acetate as mobile phase, at a flow rate of 0.2 mL x min(-1). Mass spectra were acquired in positive ion mode. Identification and structural elucidation of the components in FZHY and dosed serum were performed by comparing their retention time and MSn spectra with those of reference compounds and reported data in the literatures.

RESULTSchisandrin, schisandrol B, schisantherin A and schisandrin B were found in FZHY and dosed serum, but schisandrin C and deoxyschizandrin were only found in FZHY.

CONCLUSIONSchisandrin, schisandrol B, schisantherin A and schisandrin B can be directly absorbed into the blood after oral administration of FZHY, and the four lignans components from Schisandra chineisis might play a key role as the ingredient basement of FZHY for anti-liver fibrosis.

Administration, Oral ; Animals ; Blood Chemical Analysis ; methods ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Lignans ; blood ; Male ; Mass Spectrometry ; methods ; Rats ; Rats, Wistar

Objective To explore a promising system for tumor CD 44 receptor-targeted imaging and to investigate their physic-chemical properties and targeting effect on CD 44 abundant cancer cells in vitro.Methods The superparamagnetic iron oxide ( SPIO) nanoparticles were prepared by a coprecipitation in alkaline media starting from a mixed of the ferrous and ferric solution.And then the surface of the SPIO nanoparticles were modified with APTMS by a reaction with the hydroxyl groups.Finally, the hyaluronan-modified SPIO ( SPIO-HA) nanoparticles were prepared.Control and experimental groups were established after adding SPIO or SPIO-HA as agents respectively.Transmission electron microscopy ( TEM) and particle size analyzer were used to measure these nanoparticle sizes and the hydrodynamic diameters.Thermogravimetric analysis ( TGA) was carried out to evaluate the HA-content on the surface of SPIO-HA.The MRI T2 ralaxivities (1/T2 ) of the two groups at different Fe concentrations (0.09, 0.18, 0.27, 0.36, 0.45 mmol/L ) were measured on a 3.0T MR system.HepG2 cells and HL7702 cells were used for assessment of cells viability by methyl thiazolyl tetrazolium ( MTT ) assay.Prussian blue staining , immunoassay fluorescence image and flow cytometry were carried out to determine the targeted cellular uptake of SPIO-HA nanoparticles.MRI were performed to show the MR T 2 value changes after incubating with HepG2 cancer cells by using T 2 WI sequences at a clinical 3.0 T MR system.One-way analysis of variance was performed to determine significant changes in MR T 2 values of blank control , SPIO-HA and SPIO groups.Results The SPIO-HA and SPIO NPs were fairly homogeneous with an average core size of 18.2 and 22.4 nm, hydrodynamic diameter of 91.1 and 103.2 nm, Zeta potential of (-45.00 ±0.86) mV and (-18.50 ±0.73) mV, and magnetic relaxivity of 0.212 ×106 M-1 · s-1 and 0.191 ×106 M-1 · s-1.Based on the TGA data , HA accounted for 24%weight of each SPIO-HA.The internalization of the SPIO-HA was confirmed by prussian blue staining , while the cells showed no obvious blue stains with SPIO , incubation of SPIO-HA with tumor cells led to blue color inside the cells.After that, we examined cancer cell binding of FITC-SPIO-HA by immunoassay fluorescence image and flow cytometry.The green fluorescence resulting from FITC-SPIO-HA was observed inside the cells in both the cytoplasm and the plasmalemma.Tumor cells treated with SPIO-HA exhibited higher fluorescence signals with 7.97-fold enhancement observed for HepG 2 cells over control particles.In vitro MR, mean T2 values of blank control , SPIO and SPIO-HA groups were ( 115.20 ±0.36 ), ( 115.07 ±0.81 ) and ( 21.67 ±0.21 ) ms, respectively.There was significant difference among those three groups (F=31 703.339,P<0.01), MR T2 values of HepG2 cells treated with the SPIO-HA NPs were lower than blank and SPIO group.In comparison, SPIO did not generate any MRI signal changes compared with blank group.Conclusion The tumor CD44 receptor-targeted MR molecular probe SPIO-HA had a good physic-chemical property and well targeted HepG2 cells.