ObjectiveTo investigate the effects of astragalin on pulmonary edema after acute cervical spinal cord injury in rats.MethodsA total of 200 adult Wistar rats weighing 240-250 g were randomly divided into five groups:astragalin group,low concentration astragalin group,physiological saline group,blank group and sham group,with 40 rats in each group.The rats with cervical spinal cord injury were induced at C7 by modified Allen' s method,with the dropping weight of 10 × 2.5 g · cm.In the sham group,the laminas were removed only,leaving spinal cord at the C7 intact.Each group was further divided into four time points:24 hours,3 days,1 week and 2 weeks after the modeling,with 10 rats in each time point,according to the specimen collection time.Rats were sacrificed at different time points to observe the pathological change of the lung tissue using optical microscope,measure the lung wet/dry weight ratio (W/D) and protein concentrations of the serum and bronchoalveolar lavage fluid and calculate the lung W/D and lung permeability index (LPI).ResultsAt the same instant,the W/D and LPI in the astragalin group and low concentration astragalin group were lower than those in physiological saline group and blank group,with the lowest value in the astragalin group at day 3 after injury ( P ＜0.05 ).ConclusionsRats with acute cervical spinal cord injury may cause pulmonary edema,which can be efficiently alleviated through early use of astragalin.

BACKGROUND: Primate is the best animal to establish a model of spinal implants. However, ethics and cost limit its application. Mixed-breed dogs have similar anatomic structures as human. Moreover, it is easy to obtain with low cost, so it may replace primate to serve as models.OBJECTIVE: To determine the feasibility of canine lumbar spine to establish the anterior interfixation model following measurements of related anatomic data.METHODS: A total of 9 adult healthy dogs were selected. The transversal diameter, sagittal diameter and height of vertebral body and disc were measured, and the artificial vertebral body replacement was performed. Bone graft fusion was observed. RESULTS AND CONCLUSION: The transversal diameter, sagittal diameter and height of vertebral body and disc increasedgradually from L_(1) to L_(7), and the height was greatly exceeded its sagittal diameter. All dogs survived, but one was paraplegia. Allthe others stood and acted in 12 h to 72 h after operation. The fusion effect was proved to be good by imageology and histology. The establishment of models was simple and cost-effective, and the biocompatibility of bone tissues and implants, as well as thebone tissue ingrowth can be observed. The lumbar spine of dog can be used as an anterior interfixation model in vitro test.

Objective To establish a real-time fluorescence quantitative methylation assay to investigate the methylation status of GSH-sulphur-transferase P1(GSTP1) gene promoter region in hepatocellular carcinoma(HCC) and to investigate whether which can be used as the early diagnostic indicator of HCC .Methods Ninety-five serum samples were collected from 40 patients with HCC ,30 patients with liver cirrhosis and 25 individuals with healthy physical examination as controls .The methylation level of GSTP1 gene in these serum samples were quantitatively determined by using the real-time fluorescence quantitative methylated spe-cific PCR technique .The receiver-operation characteristic(ROC) curves were adopted to evaluate its diagnostic value for HCC .Re-sults The methylation quantitative level of GSTP1 gene in HCC serum was significantly higher than that in the healthy controls (P<0 .05) .The ROC curve analysis demonstrated that the methylation quantitative analysis of GSTP1 gene could efficiently distin-guish HCC and cirrhosis from healthy controls (AUC=0 .8641) .With the methylation rate of 2% as the critical value for diagno-sing HCC ,its diagnostic specificity was 87 .5% ,the sensitivity was 69 .6% ;the combination detection of serum GSTP1 gene methy-lation and serum AFP could increase the detection rate of HCC to 75% .Conclusion The real-time fluorescence quantitative methyl-ation assay can accurately quantify the methylation level of serum GSTP1 gene ,which has certain application value for the early di-agnosis of HCC .

We predicted the anti-hepatitis B virus (HBV) active components and mechanism of Salvia miltiorrhiza based on network pharmacology. The active components of S. miltiorrhiza were obtained through TCMSP, PubChem database and literature research. The potential targets of the active components and HBV infection were predicted by SwissTargetPrediction and GeneCards databases, respectively. The protein-protein interaction (PPI) network was constructed by String database. Cytoscape software was adopted to construct a visual network of active component-disease target and perform topological analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed using DAVID platform. The molecular docking of key components and core targets was carried out by AutoDock Vina software. We screened out a total of 38 active components and 178 disease-component overlapping targets. Enrichment analyses obtained 405 related GO items and 68 signaling pathways, such as T/B cell receptor signaling pathways, PI3K/AKT signaling pathway, and mTOR signaling pathway. According to the results of molecular docking, most characteristic components of S. miltiorrhiza (miltionone Ⅱ, miltirone, protocatechuic acid, lithospermic acid, protocatechualdehyde) showed good affinity with the key targets (PIK3CA, APP, STAT3,AKT1 and mTOR). Furthermore, the anti-HBV activity of lithospermic acid, the representative active component of S. miltiorrhiza, and its regulation on PI3K/AKT and mTOR signaling pathways were investigated in an HBV replicating mouse model. Animal welfare and experimental procedures follow the regulations of the Animal Ethics and Welfare Committee of Hubei University. The results showed that lithospermic acid significantly inhibited HBV DNA replication, reduced serum HBsAg and HBeAg levels, and decreased the phosphorylation protein expression levels of AKT and mTOR in liver, indicating that lithospermic acid might exert the anti-HBV activity by regulating PI3K/AKT and mTOR signaling pathways.

Aim To study inhibitory effect of 5-fluorouracil encapsulated by galactosylceramide liposomes (5-Fu-GCL)on 5-Fu-resistent HepG_2 cells and its mechanisms. Methods Inhibitory effect of 5-Fu-GCL on established model of 5-Fu-resistant HepG_2 cells was assessed with MTT assay in vitro. The concentration-time course of 5-Fu-GCL in intracellular fluid was detected with high performance liquid chromatography (HPLC). Thymidylic acid synthase (TS) expression was observed with immunohistochemical method,and NO content was determined with chemical method. Results Obvious inhibitory effects of 5-Fu-GCL (75,150,300,600,1200?mol?L-1) on 5-Fu-resistant HepG_2 cells were observed with IC_ 50 of 158.6 ?mol?L-1,far lower than that of free 5-Fu (400.9 ?mol?L-1). 5-Fu-GCL (300 ?mol?L-1) inhibited 5-Fu-resistant HepG_2 cells in a time-dependent manner,and the inhibitory effect of 5-Fu-GCL was stronger than that of free 5-Fu during 12～48 h. Compared with free 5-Fu,5-Fu-GCL (300 ?mol?L-1) increased the content of intracellular fluid in 5-Fu-resistant HepG_2 cells. 5-Fu-GCL(62.5,300,1200 ?mol?L-1) not only inhibited the expression of TS,but also increased the production of NO in 5-Fu-resistant HepG_2 cells,and these effects of 5-Fu-GCL(300,1200 ?mol?L-1) were stronger than those of free 5-Fu. Conclusion 5-Fu-GCL has inhibitory effect on 5-Fu-resistant HepG_2 cells. The effect may be related to the increased concentration of 5-Fu-GCL in intracellular fluid,inhibited expression of TS and increased production of NO.

Objective To evaluate the clinical efficacy of multiple stents placement in the management of hilar cholangiocarcinoma,especially in the complex cases of which the hepatic ducts are invaded.Methods Forty-five consecutive patients with hilar cholangiocarcinoma were treated with percutaneous transhepatic placement of two or three self-expandable metallic endoprostheses.The cause of hilar obstructions in these patients were all cholangiocarcinoma,including Bismuth classification type Ⅱ(n 12 ),Ⅲa(n 17),Ⅲb(n 10),and Ⅳ(n 6).Two or 3 stents were placed in the configuration of T,Y or X over the strictures.Results Stent placement with 2 or 3 endoprostheses was successful in all patients.All patients showed significant decrease in serum bilirubin level.The mortality rate within 30 days of stent placement was 2.2%(1/45).The mean survival and stent patency times were 215.3 d(26— 516 d)and 181.5 d(26—473 d),respectively.Conclusion Deploying of multiple metallic stents is an effective method to treat complex hilar cholangiocarcinoma,especially for the cases of which hepatic ducts are invaded:the henatic ducts should be drained as much as nossible.

Objective To study the characteristics of the transcranial electric stimulation(TES)-elicited motor evoked potentials(MEP)recorded from different points of the diaphragm,and to determine the optimal record- ing site in the diaphragm.Methods One EEG electrode was placed subcutaneously in the midline of the skull as stimulation electrode and another EEG electrode was inserted into hard palate submucously as cathode in 30 male rats.And single square wave electrical pulse was used for stimulation.The stimulation intensity was 15 mA,the du- ration of the pulse was 0.2 ms,and the stimulating interval was 200 ms.The concentric needle electrode were insert- ed into the following eight sites:the medial,intermediate,lateral crura of the diaphragm,the junction of the posterior axilla line and the inferior border of the eleventh rib,the junction of the anterior axilla line and the inferior border of the ninth rib,the junction of the intermediate line of the clavicle and the inferior border of the eighth rib,the junction of the para-sternal line and the inferior border of the seventh rib,the pars sternalis.The MEP was recorded from each point,respectively.The optimal sites of the diaphragm MEP were found and recorded.Results The MEPs were re- corded from each of the 8 sites of the diaphragm in all the rats after TES.There was no statistically significant differ- ence among the latencies of every site.But the amplitude varied from 6.08?0.46 mV in maximum along the inferior border of the ninth rib at the anterior line of the axilla to the minimum of O.91?0.03 mV at the pars sternalis. Conclusion It is safer to insert the needle along the inferior border of the ninth rib at the anterior line of the axilla, because there was certain distant from the folding part of the pleura and the amplitude of the recorded wave at this point is maximal.

Objective To investigate the features of chronic prostatitis during puberty(CPP)and the effects of pelvic floor biofeedback therapy.Methods Totally,25 CPP children (mean age,16 years) and 15 children (mean age,16 years) with normal lower urinary tract as controls were included.In CPP group,NIH-CPSI scores were evaluated,expressed prostatic secretions (EPS) were examined,and bacterial culture was done;and CPP patients were categorized based on the definitions of NIH types.In both groups, urodynamic examination was performed,including evaluation of uroflow curve,maximum flow rate (Q_(max)), post-voiding residual urine (PVR),detrusor-sphincter dyssynergia (DSD),maximum detrusor pressure (P_(det,max))and maximum urethral closure pressure (MUCP).CPP patients underwent biofeedback therapy, and 10 weeks later the effects were assessed.Results In CPP group,NIH typing showedⅡ,ⅢA andⅢB in 1,3 and 21 cases,respectively.Before treatment in CPP and control groups,the incidence of staccato voiding (20 cases vs 1 case),DSD (22 cases vs 1 case),Q_(max)(10.7?3.7 vs 15.0?4.3ml/s),PVR (7.7?4.1vs 3.2?2.6ml),P_(det,max)(115.1?33.6vs 76.8?16.6cm H_2O)and MUCP(176.5?45.7 vs 86.2?28.5cm H_2O)all showed significant differences between the 2 groups(P＜0.05).In CPP group,the differences in pain(4.6?2.2 vs 2.1?1.6),urination (7.9?2.0vs 2.2?1.7),life impact (9.4?2.2vs 2.6?2.1)and total scores(22.0?5.2vs 7.0?4.2) of NIH-CPSI and Q_(max)(10.7?3.7 vs 14.9?5.6) between pre-and post-biofeedback were significant (P＜0.05).Conclusions The main type of CPP is categoryⅢB.The primary symptom is voiding disorder,which leads to greater psychological stress in patients.Children with CPP have pelvic floor dysfunctions and multiple abnormal urodynamic param- eters.The short-term effect of biofeedback strategies for CPP is satisfactory.

Objective To study the relationship among apoptosis,NF-KB activation and uPA expres- sion in human colon carcinoma cell line HCTll6 induced by 5-fluorouracil,and to observe the effect of in- hibiting activity of NF-KB by PDTC on apoptosis as well as expression of uPA.Methods Cell apoptosis was analysed by Annexin V-FITC.Fluctuation of NF-KB and uPA was detected by semi-quantitative immuno- histochemistry.Results 5-fluorouracil could induce apoptosis and activate NF-KB.PDTC could significantly increase the apoptosis and suppress the activation of NF-KB induced by 5-fluorouracil.There was a positive correlation between the changes of uPA and NF-KB.Conclusion 5-fluorouracil could induce apoptosis,ac- tivate NF-KB and up-regulate expression of uPA of HCT116 cells.The mechanism of enhanced apoptosis by PDTC may be related to suppressing activation of NF-?B and down-regulating expression of uPA.

Humans ; Male ; Middle Aged ; Pacemaker, Artificial ; adverse effects ; Tricuspid Valve Stenosis ; etiology