AIM:To construct eukaryotic expression vector expressing double shRNA sections targeting Survivin gene.METHODS: Eukaryotic expression vector expressing double shRNA sections targeting Survivin gene were designed and chemically synthesized.They were directionally inserted into plasmid pGenesil-1 with respectively U6 promoter and termination code,the common green fluorescence protein(EGFP) gene and Neo gene. In this way,the vector of pGenesil-1 shRNA containing 2 sections of Survivin shRNA were constructed and they were transfected into the pancreatic cancer cell Bx-PC3.Transfection was detected by fluorescence microscope.The inhibition expression of Survivin mRNA was measured by RT-PCR.RESULTS: HE1 and HE2 plasmids were identified by the biocatalyst cut which confirmed the exactitude and were analyzed by the sequence analysis which verified the perfect clone plasmid inserted by them.CONCLUSION: A eukaryotic expression vector of double short hairpin RNA for Survivin gene is successfully constructed.The pancreatic cancer cells Bx-PC3 succeed to be transfected and expression of Survivin mRNA is inhibited obviously.

OBJECTIVETo observe the distribution of the blood vessels in the integument tissue of the channel area of legs.

METHODSThe integument tissue of the lower limbs in the 12 cadavers were dissected with macro-and micro-dissection, radiographical technique of systemic artery and technique of image pattern analysis to observe and analyze the origins, branches and anastomoses in the integument tissues along the channels of legs.

RESULTSThe distributional density of the blood vessels in the integument tissues of legs along the channel area of the three-yin meridians of the foot, the Gallbladder Meridian, and the Urinary Bladder Meridian was higher than that in the other parts. They formed an obvious nutrient vascular chain on the arteriogram. The distributional density in the channel area of the Stomach Meridian was not obviously increased and the obvious nutrient vascular chain could not be seen.

CONCLUSIONAn obvious nutrient vascular chain is formed in the integument tissue along the channel area of legs except the Stomach Meridian.

Blood Vessels ; anatomy & histology ; Humans ; Leg ; blood supply ; Meridians

OBJECTIVETo screen and identify proteins that interact with hepatitis C virus NS3 by means of T7-phage display system.

METHODSHepatitis C virus NS3 was expressed by prokaryotic expression and used as a selected molecule to biopan the T7 select human liver cDNA library; the selected positive clones were identified using DNA sequencing and analyzed with BLAST program in GenBank.

RESULTSAfter BLAST analysis in all the positive clones, the proteins which interacted with the hepatitis C virus NS3 were found to be serpin peptidase inhibitor, clade A, member 1 (SERPINA1) and cyclophilin-LC.

CONCLUSIONT7-phage display system is a convenient, rapid and effective method for screening interacting proteins. The proteins thus selected will provide an important means for studying the pathogenesis and carcinogenesis of HCV.

Cell Line ; Gene Library ; Hepacivirus ; metabolism ; Humans ; Peptide Library ; Protein Interaction Mapping ; methods ; Viral Fusion Proteins ; genetics ; isolation & purification ; metabolism ; Viral Nonstructural Proteins ; genetics ; metabolism

OBJECTIVETo investigate the expression profiles of serum Golgi protein-73 (GP73) in liver cirrhosis and primary hepatic carcinoma (PHC) and determine its clinical value for differential diagnosis.

METHODSSerum protein expressions of GP73 and alpha-fetoprotein (AFP) were detected by enzyme-linked immunosorbent assay and chemiluminescence assay, respectively, in patients with PHC (n=80), liver cirrhosis (n=65), and healthy controls (n=50). Inter-group changes were assessed by Kruskal-Wallis test, and significance of these differences was assessed by Mann-Whitney test. A receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic efficiency and determine the cut-off values for GP73 and AFP. Sensitivity and specificity were compared by the Chi-squared test. Correlation between serum GP73 expression and clinical parameters was determined by Spearman's rank correlation analysis.

RESULTSThe PHC group showed significantly higher serum GP73 (282.0 mug/L) than the liver cirrhosis group (211.8 mug/L) and control group (58.3 mug/L) (H = 93.30, P less than 0.01). For differential diagnosis of PHC and liver cirrhosis, the cut-off value was 318.1 mug/L for GP73 and 13.4 mug/L for AFP. Sensitivity of GP73 was lower than AFP (45% (36/80) vs. 65% (52/80); X2 = 8.02, P less than 0.05). Specificity of GP73 was lower than AFP but no significance was found (83.1% (54/65) vs. 87.7% (57/65); X2=0.27, P more than 0.05). The areas under the ROC curves were not significantly different between GP73 and AFP (0.65 (95% confidence interval (CI): 0.54~0.72) vs. 0.75 (95% CI: 0.67~0.83); Z = 1.88, P more than 0.05). The area under the ROC curves increased but not significantly (0.80 (95% CI: 0.73~0.88) vs. 0.75 (95% CI: 0.67~0.83); Z=2.61, P more than 0.05). Serum GP73 was correlated with liver cirrhosis (r=0.27), vascular invasion (r=0.29), and TNM staging (r=0.27) (all P less than 0.05), but not with sex (r=0.13), age (r=0.10), enhanced AFP (> 13.4 mug/L; r=0.03), tumor size (r=0.18), or distant metastasis (r=0.04), all P less than 0.05.

CONCLUSIONSerum GP73 and AFP have comparable diagnostic efficiency, but the sensitivity of AFP is superior for differential diagnosis of liver cirrhosis and primary hepatic carcinoma. Elevated serum GP73 may be correlated with liver tumor load and aggressiveness.

Adult ; Aged ; Carcinoma, Hepatocellular ; diagnosis ; Case-Control Studies ; Diagnosis, Differential ; Female ; Humans ; Liver Cirrhosis ; diagnosis ; Liver Neoplasms ; diagnosis ; Male ; Membrane Proteins ; blood ; Middle Aged ; Sensitivity and Specificity ; Transcriptome ; alpha-Fetoproteins ; metabolism

OBJECTIVETo systematically review the effectiveness and safety of open and laparoscopic surgeries for treatment of adrenal tumors.

METHODSThe online databases including CNKI, PUBMED, SinoMed, EBSCO, Springerlink, WanFang Data, and VIP were searched for clinical trials published from 1999 to 2016. A meta-analysis was performed using RevMan 5.2 software.

RESULTSA total of 2340 patients in 25 trials were included. The results of meta-analysis showed that laparoscopic surgery was better than open surgery in terms of intestinal function recovery time (OR=-0.96, 95%CI [-1.22, -0.70] P<0.000 01), hospitalization time (OR=-3.48, 95%CI [-4.13, -2.78], P<0.000 01), complications (OR=0.22, 95%CI [0.14, 0.35], P<0.0001), and volume of blood loss (OR=-104.77, 95%CI [-138.95, -70.60], P<0.000 01). There was no significant difference in the surgery cost between open and laparoscopic surgeries.

CONCLUSIONLaparoscopic surgery is superior to open surgery for treatment of adrenal tumors for shorter intestinal function recovery time, surgery duration, and hospitalization time and less complications and blood loss.

OBJECTIVEFabricate series of the controllable degradation coral-hydroxyapatite.

METHODSThe natural coral undergo a chemical reaction with (NH4)2 HPO4 at high temperature and pressure for different time-lengths. After getting the products, the components and the special structures were analyzed. Observe the biologic degradation of the reaction products and analyze the metal elements and their contents. Haemolysis tests, cytotoxity tests and bone compatibility tests were performed to assess the biocompatibility of the products.

RESULTSWhen hydrothermal reactions happened under different conditions, the different gradients of CaCO3/hydroxyapatite materials were produced. These types of materials kept the characteristic of interconnected micro-porous network structures. A thin layer of compact material can be seen on the surface of its trabecula ultra-micro structure. The SCHA-200R has a good biocompatibility.

CONCLUSIONSGradient HA (SCHA-200R) materials can be formed by adjusting the same temperature, same pressure and different time-length of the reaction. This kind of gradient material keeps the quality of micro-porous network structures. The SCHA-200R is a potential candidate scaffold for bone tissue engineering.

Absorbable Implants ; Animals ; Anthozoa ; chemistry ; Bone Substitutes ; Durapatite ; chemical synthesis ; Male ; Materials Testing ; Rabbits ; Tissue Engineering ; methods

OBJECTIVETo explore the effect of electroacupuncture (EA) on the pathomorphology of the sciatic nerve and the role of P2X3 receptors in EA analgesia.

METHODSThe chronic constriction injury (CCI) model was adopted in this study. A total of 32 rats were randomly divided into four groups: sham CCI, CCI, CCI plus contralateral EA (CCI + conEA) and CCI plus ipsilateral EA (CCI + ipsEA). Mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were measured. EA began at day 7 after the CCI operation and was applied to the Zusanli (ST 36) and Yanglingquan acupoints (GB 34). At day 14, the pathomorphologic changes of the operated sciatic nerve were demonstrated by hematoxylin and eosin staining. In addition, dorsal root ganglion (DRG) neurons isolated from rats were examined by electrophysiological recording to determine if the P2X3 receptor agonists, adenosine 5'-triphosphate disodium (ATP) and α,β-methylen-ATP (α,β-meATP) evoked inward currents.

RESULTSPain thresholds in the CCI group were obviously decreased post CCI surgery (P<0.01). In the EA groups, thermal and mechanical threshold values were increased after the last EA treatment (P<0.05, P<0.01). There was no significant difference in light microscopic examination among the four groups (P>0.05). Current amplitude after application of ATP and α,β-meATP in DRG neurons were much larger in the CCI group compared to those obtained in sham CCI (P<0.05). ATP and α, β-meATP invoked amplitudes in the CCI + EA groups were reduced. There was no signififi cant difference between the CCI + conEA group and the CCI + ipsEA group (P>0.05).

CONCLUSIONEA analgesia may be mediated by decreasing the response of P2X3 receptors to the agonists ATP and α,β-meATP in the DRG of rats with CCI. No pathological changes of the sciatic nerve of rats were observed after EA treatment.

Adenosine Triphosphate ; analogs & derivatives ; pharmacology ; Animals ; Constriction, Pathologic ; Electroacupuncture ; Ganglia, Spinal ; drug effects ; metabolism ; pathology ; Hyperalgesia ; pathology ; Ion Channel Gating ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Reaction Time ; drug effects ; Receptors, Purinergic P2X3 ; metabolism ; Sciatic Nerve ; injuries ; metabolism ; pathology ; Staining and Labeling

OBJECTIVETo study the differential gene expression profiling of rats exposed to silica using the normal rats as control.

METHODSAnimal models were established using intratracheal injection of the lung and 22 107 genes were screened in the differential expression profiling of silicosis by using oligonucleotide bead array. Differential expression profiling data were analyzed by using DAVID bioinformation software.

RESULTSTotally 1567 differentially expressed genes were identified in lungs of silica exposed rats including 765 up-regulated genes and 802 down-regulated genes as compared to the normal controls. Among 406 annotated genes in KEGG pathways, 204 genes and 11 pathways were up-regulated as well as 202 genes and 3 pathways were down-regulated in silica exposed rats.

CONCLUSIONAll 1567 genes are involved in the formation of silicosis. The differential gene expression profile of silicosis describes the general changes in the gene expressions in silicosis at transcriptional level. Further analysis of the identified genes might help reveal the molecular mechanism of pulmonary fibrosis induced by silica.

Animals ; Disease Models, Animal ; Gene Expression Profiling ; Lung ; metabolism ; pathology ; Male ; Oligonucleotide Array Sequence Analysis ; Pulmonary Fibrosis ; genetics ; metabolism ; Rats ; Rats, Wistar ; Silicosis ; genetics ; metabolism ; pathology

Adult ; Foreign Bodies ; diagnostic imaging ; Humans ; Male ; Radiography ; Stents ; adverse effects ; Ureter

OBJECTIVETo study the gene mutation in a patient with multiple exostoses, identify the disease-causing gene mutation.

METHODSPolymerase chain reaction and DNA sequencing were used to screen the EXT1 or EXT2 gene mutation, while mismatch primer amplification and restriction endonuclease digestion were performed to confirm the mutation.

RESULTSBy DNA sequencing, a mutation in the seventh intron was detected and located at 26 bp of 3' splice site upstream in EXT1 gene, which was unreported before. Mismatch primer amplification and restriction fragment length polymorphism analysis suggested that this mutation was not detected in the normal control.

CONCLUSIONThe mutation 1633-26(C-->A) may be the disease-causing mutation in this patient with multiple exostoses.

DNA Mutational Analysis ; Exostoses, Multiple Hereditary ; genetics ; Female ; Humans ; Mutation ; N-Acetylglucosaminyltransferases ; genetics ; Young Adult