BackgroundWhether the peeling of the inner limiting membrane (ILMP) increase the closure rate of idiopathic macular hole is still in controversy.Some ophthalmologist recommend vitrectomy combined with inner limiting membrane peeling for the treatment of idiopathic macular hole.However,the removal of ILMP is difficult because of its similar appearance to adjacent tissues.Objective This study was to investigate the efficacy of triamcinolone acetonide(TA) and indocyanine green(ICG) double staining-assisted vitrectomy combined with ILM peeling during the surgery.Methods A consecutive case- observational study was designed.The standardized vitrectomy was performed in 25 eye of 23 cases with IMH.During the vitrectomy,TA and ICG were injected into posterior pole vitreous to visualize and assist the ILM peeling.The dying effectiveness was observed,and the closure rate of macular hole,visual acuity,intraocular pressure and complications were evaluated after surgery.Written informed consent was obtained from each patient prior to operation.Results Posterior vitreous cortex and ILM were visible and the residual vitreous and cortex were removed clearly after dying of TA and ICG in all the 25 eyes.During the following-up duration of 3-8 months,the completely anatomical reattachment of the macular area was in 22 eyes ( 88.0％ ) and partially reattachment in 3 eyes( 12.0％ ).The best corrected vision was 0.07-0.60 in all of the operated eyes 2 months after surgery.Conclusions TA and ICG- assisted vitrectomy combined with ILM peeling appears to be a safe and effective method for IMH repair.

Background: Previous study has found that ursolic acid (UA) inhibited the proliferation of gastric cancer cells by the down-regulation of cyclooxygenase-2 (COX-2) expression.However,its molecular mechanism is not fully clear.Aims: To investigate the role of adenosine monophosphate-activated protein kinase (AMPK)/signal transducer and activator of transcription 3 (STAT3)/COX-2 signaling pathway in UA-mediated inhibition of gastric cancer cells proliferation.Methods: AMPK-pLVX,AMPK-shRNA,STAT3-pLVX,STAT3-shRNA plasmids were constructed,and then were transfected into human gastric cancer cell lines SGC-7901 and MKN-45,respectively.Gastric cancer cells were cultured with different concentrations of UA for different times.The expressions of phosphorylated AMPK (p-AMPK),phosphorylated STAT3 (p-STAT3) and COX-2 were measured by Western blotting,and cell proliferation was detected by CCK-8 assay.Results: UA dose-and time-dependently increased p-AMPK expression,inhibited p-STAT3 and COX-2 expressions in SGC-7901 and MKN-45 cells.Knockdown of AMPK blocked UA-induced inhibition of STAT3 phosphorylation and COX-2 expression.Overexpression of STAT3 blocked UA-induced down-regulation of COX-2 expression.Knockdown of AMPK and overexpression of STAT3 blocked UA-induced inhibition of proliferation of gastric cancer cells.Conclusions: UA may inhibit the proliferation of gastric cancer cells via down-regulation of COX-2 expression through AMPK/STAT3 pathway.

Objective To evaluate the diagnostic value of detection of IgM antibodies to EV71-infection patients,and compared characterisation of RT-PCR,IgM capture ELISA and neutralization test.Methods Virus RNA,neutralization titer and IgM antibody in 115 EV71-infection patients were detected by EV71 real-time RT-PCR kit( EV71-PCR kit),neutralization test,and EV71 IgM-capture ELISA kit (EV71-IgM kit),respectively.Results Using EV71-IgM kit,the detection rate was 80.9％ (93/115,95％ CI:72.5-87.6) among the 115 EV71-infection patients,and was 2.6％ among the 228 healthy children.Simultaneously,sera collected after 1-2 day of disease onset showed an IgM positivity of 70.4％ (38/ 54).The positive rate of EV71-PCR among these patients was 82.6％ (95/115,95％ CI:74.4-89.0),so there was no statistically significant differences between it and EV71-IgM kit.In addition,the detection rate in EV71-infection patients could increase to 92.2％ by combined detection of EV71-PCR and EV71-IgM kit.Conclusion EV71-IgM kit was a rapid and valuable way for the early diagnosis of EV71 infection,and could significantly improve detection rate for EV71 infection by combining with EV71-PCR kit.

OBJECTIVETo explore the molecular mechanism of dutasteride inhibiting fertility by studying its effects on the expressions of the epididymal epithelial junction proteins Claudin1 and β-catenin in rats.

METHODSSixteen 3-month-old SD male rats were equally divided into an experimental and a negative control group to be treated intragastrically with dutasteride at 40 mg/kg per day and the same dose of solvent, respectively, for 14 consecutive days. Then, the sperm motility and morphology of the rats were detected by computer-assisted sperm analysis, the serum levels of testosterone (T) and dihydrotestosterone (DHT) measured by ELISA, changes in the tight junction of epididymal cells observed under the transmission electron microscope, the protein and gene expressions of Claudin1 and β-catenin determined by RT-PCR and immunohistochemistry, and the conception rate of the mated female rats calculated.

RESULTSDutasteride significantly suppressed the serum DHT level, sperm motility, and fertility of the rats (P <0.05). Interspaces between epididymal epithelial cell tight junctions were observed, the volume of epididymal fluid obviously increased, and the expressions of Claudin1 and β-catenin gene and protein remarkably downregulated in the experimental rats (P <0.05).

CONCLUSIONDutasteride can significantly inhibit the fertility of male rats by reducing the serum DHT level, suppressing Claudin1 and β-catenin expressions, and damaging epididymal epithelial cell junctions.

Animals ; Azasteroids ; pharmacology ; Claudin-1 ; metabolism ; Dihydrotestosterone ; blood ; Dutasteride ; Epididymis ; drug effects ; metabolism ; Female ; Fertility ; drug effects ; Humans ; Intercellular Junctions ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Sperm Motility ; drug effects ; Testosterone ; blood ; Urological Agents ; pharmacology ; beta Catenin ; metabolism

To further explore the mechanism of congenital pyrimidine 5'-nuleotidase I (P5'N-I) deficiency, on the basis of purification of the protein, the molecular weight and amino acid composition were analysed by mass-spectrograph and amino-acid analyzer, microsequencing and bioinformation analysis of P5'N-I were performed after it was hydrolysed by trypsin. The results showed that three fractions were found in the purified P5'N-I and their molecular weights were 26,952.9, 55,476 and 110,938, respectively. The sequence from one of the peptide fragments was I-E-G-P-T-I-R-Q-I-E. The homologous sequence was not found after comparision with the ten-amino-acid sequence in GenBank by blast procedure. Amino acid analysis indicated that P5'N-I was composed of 18 amino acids at least, and 243 amino acid residues. In conclusion, the enzyme might be an allosteric enzyme, there might be homologous dimer or tetramer in physiological status of normal human erythrocyte, the microsequence could be designed as the probe for fishing the genes of interest. The composition of amino acid might be an important information in determination of its protein primary structure.
5'-Nucleotidase ; blood ; chemistry ; isolation & purification ; Amino Acid Sequence ; Amino Acids ; analysis ; Chromatography, High Pressure Liquid ; Electrophoresis, Polyacrylamide Gel ; Erythrocytes ; enzymology ; Humans ; Mass Spectrometry ; Molecular Weight ; Peptide Fragments ; chemistry ; Sequence Analysis, Protein

OBJECTIVETo prepare (99m)Tc-labeled Anti-VEGF mAb 5-FU loaded polylactic acid nanoparticles ((99m)Tc-Ab-5-FU-NPs) and investigate its biodistribution in human gastric carcinoma xenografts.

METHODS(99m)Tc-Ab-5-FU-NPs were prepared by labeling Ab-5-FU-NPs with (99m)Tc using improved Schwarz method. After isolation of (99m)Tc-Ab-5-FU-NPs using SephadexG250 column, the labeling ratio and radiochemical purity were determined using chromatography. The immunocompetence of (99m)Tc- Ab-5-FU-NPs was detected by ELISA and immunohistochemistry. (99m)Tc-Ab-5-FU-NPs were then injected via the tail vein into SCID mice bearing human gastric carcinoma, and (99m)Tc labeled mice-derived monoclonal IgG loaded polylactic acid nanoparticles were used as the control, followed by radioimmunoscintigraphic imaging at 2 and 6 h. The radioactive count and radioactive ratio of the tumor and non-tumor tissue (T/NT) in the animal models were calculated using ROI technique. After imaging at 24 h, SCID mice were sacrificed and the radioactive distribution, the %ID/g, as well as the T/NT radioactive ratio were examined, respectively. The concentrations of 5-FU in the tumor and blood were also detected using HPLC method.

RESULTSThe labeling ratio of (99m)Tc-Ab-5-FU-NPs was 90%-95%. (99m)Tc-Ab-5-FU-NPs were detected in the tumor tissues by radioimmunoimaging 2 h after the injection. ID%/g in the tumor tissues at 2 and 6 h were both significantly higher than that of the control group. Both the ID%/g in tumor tissues and radioactive ratio of tumor and blood at 6 h were higher than those at 2 h, and the concentration of 5-FU in experimental group increased continuously with time and was significantly higher than that in control group.

CONCLUSIONS(99m)Tc-Ab-5-FU-NPs prepared in this study can meet the demands of radioimmunoimaging, and the anti-VEGF monoclonal antibody possesses reliable immune targeting ability. Six hours after injection, (99m)Tc-Ab-5-FU-NPs can specifically accumulate in the tumor tissues in human gastric carcinoma xenografts at high concentration.

Animals ; Antibodies, Monoclonal ; chemistry ; immunology ; Cell Line, Tumor ; Cell Transformation, Neoplastic ; Female ; Fluorouracil ; blood ; chemistry ; pharmacokinetics ; Humans ; Lactic Acid ; chemistry ; Male ; Mice ; Mice, SCID ; Nanoparticles ; Polyesters ; Polymers ; chemistry ; Radioimmunotherapy ; Stomach Neoplasms ; blood ; metabolism ; pathology ; radiotherapy ; Technetium ; chemistry ; Vascular Endothelial Growth Factor A ; immunology

OBJECTIVETo study the bone state with ultimate stress by examining biomechanical distribution of upper femur in Chinese, in order to accumulate more experiences for clinical work.

METHODSTotally 60 Chinese femurs from fresh cadavers were randomly divided into two groups. All the femurs were cleaned, and the body age ranged from 36 to 72 years old, averaged 56.4 years, including 41 from males, and 19 from females. These two groups underwent mechanical stress and bending stress tests. Special mechanical laboratory and machines were used to get the information. Results about the loading value at each testing point under stress were collected.

RESULTSThe four faces of the upper femur suffered different stress under external forces. The bone on upper femur can tolerate more mechanical stress than bending stress. Medial and lateral region of the femur neck and the rear side of the small tuberosity section were themain position enduring the vertical stress. The rear position of the base femur neck and the small tuberosity section were the main regions enduring the bending stress. Those main positions had strong cancellous bones. The intertrochanteric fracture fixation and artificial femoral stems were designed depending on this biomechanical basis.

CONCLUSIONAccording to our experiment result, doctors need to chose more effective fixations for upper femur fracture, and femoral stems for the patients. More information should be collected by further researches.

Adult ; Aged ; Biomechanical Phenomena ; Female ; Femur ; chemistry ; Humans ; Male ; Middle Aged ; Stress, Mechanical

OBJECTIVETo investigate the relationship between the distribution of mononucleotide polymorphism of five regulation regions of alpha-IFN among HLA-DRB1*11 gene episodes and the therapeutic effects of alpha-IFN treatment in chronic hepatitis B patients.

METHODSOne hundred seven chronic hepatitis patients from Nanjing Second Hospital who were treated by alpha-IFN for 12 months and then followed at least six months without the treatment were randomly selected for this regressive analysis. They were grouped into a continuous responsive group and a non-continuous responsive group. Hepatitis B virus X interacting protein gene locus was searched in NCBI. Single nucleotide polymorphism (SNP) gene locus was detected based on a pooling sequencing method. Primer and TaqMan-MGB probes referring to different mononucleotide loci were designed respectively to detect SNP in five regulation regions of alpha-IFN. Then gene sequencing differences between the two groups were analyzed.

RESULTSAmong the 107 cases there were 30 cases (28.0%) in the continuous responsive group and 77 cases (71.9%) in the non-continuous responsive group. CT occupation rate in five regulation regions of IFN reached 18.0% in the continuous responsive group and 23.8% in the non-continuous responsive group. AG occupation rate reached 10.8% in the former group and 15.8% in the latter group. The differences in CT and AG between the two groups were significant.

CONCLUSIONSThe distribution of mononucleotide polymorphism of five regulation regions of alpha-IFN among HLA-DRB1*11 gene episodes affects the IFN anti-virus treatment. Detecting the gene distribution of mononucleotide in five regulation regions of alpha-IFN helps in predicting the therapeutic effects of alpha-IFN.

Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; DNA, Viral ; Genotype ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; genetics ; Humans ; Interferon-alpha ; therapeutic use ; Polymorphism, Single Nucleotide ; Regression Analysis ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the association of single nucleotide polymorphisms (SNP) rs22833188 and rs2833195 in TIAM1 gene with the susceptibility to Kawasaki disease (KD) and its clinical characteristic in children.

METHODSA case-control study was performed in this study. One hundred and eighty-eight children with KD and 197 normal children served as controls were enrolled. The genotypes of two SNPs rs22833188 and rs2833195 in TIAM1 gene were detected using PCR-RFLP.

RESULTSThere were no significant differences in the genotype (AA, AG and GG) and allele frequencies of SNP rs2833188 between the KD and control groups. Significant differences in the genotype (CC, GC and GG) frequency of SNP rs2833195 were noted between the KD and control groups (P=0.017). The frequency of C allele in the KD group was higher than in the control group (P=0.015). The polymorphism of SNP rs2833188 was associated with the occurrence of rash (P=0.011), and the polymorphism of SNP rs2833195 was associated with the occurrence of conjunctival hyperemia (P=0.021).

CONCLUSIONSThe polymorphism of rs2833195 in TIAM1 gene is associated with the susceptibility to KD. The polymorphisms rs2833188 and rs2833195 in TIAM1 gene may be associated with some clinical characteristics in children with KD.

Child ; Child, Preschool ; Female ; Genetic Predisposition to Disease ; Genotype ; Guanine Nucleotide Exchange Factors ; genetics ; Humans ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; genetics ; Polymorphism, Single Nucleotide ; T-Lymphoma Invasion and Metastasis-inducing Protein 1

OBJECTIVETo investigate the changes of guinea pig heart electrophysiological properties caused by increasing left ventricular preload, and to assess the effects of tetradrine on these changes.

METHODWorking model preparation of guinea pig hearts in vitro was used, and the preload of left ventricle was increased by adjusting the prefusion pressure of left atria. The changes of heart electrophysiologic parameters including monophasic action potential duration (MAPD90), monophasic action potential amplitude (MAPA), effective refractory period (ERP) and ventricular fibrillation threshold (VFT) were observed before and after altering the preload of left ventricle, and compared in the absence and presence of tetradrine, streptomycin or verapamil.

RESULTThe rising of left ventricular preload led to shortening of MAPD90, ERP, and to descent of MAPA, VFT (all P<0.01). Both Tetradrine and streptomycin inhibited these changes of heart electrophysiologic parameters caused by elevation of left ventricular afterload (all P<0.01). In contrast, verapamil had no effects on the preload-related electrophysiological changes (all P>0.05).

CONCLUSIONElectrophysiologic changes caused by increasing left ventricular preload may be inhibited by tetrandrine, through inhibition of stretch-activated ion channels.

Action Potentials ; drug effects ; Alkaloids ; isolation & purification ; pharmacology ; Animals ; Anti-Arrhythmia Agents ; isolation & purification ; pharmacology ; Benzylisoquinolines ; isolation & purification ; pharmacology ; Calcium Channel Blockers ; pharmacology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Guinea Pigs ; Heart ; physiology ; In Vitro Techniques ; Male ; Plants, Medicinal ; chemistry ; Refractory Period, Electrophysiological ; drug effects ; Stephania tetrandra ; chemistry ; Streptomycin ; pharmacology ; Ventricular Function, Left ; drug effects ; Verapamil ; pharmacology