Objective To investigate the effects of previous open nephrolithotomy on the technical features, outcomes and morbidities of subsequent percutaneous nephrolithotomy (PCNL). Methods Ninety-eight patients who underwent PCNL from January 2006 to January 2011 were selected in this study. The 34 patients of them who had previous open nephrolithotomy on the same kidney were assigned as group A, and the other 64 patients who had no previous open surgery as group B. The data of operation time, blood transfusion quantity, residual stones rate, hospitalization time and time of tube evulsion were collected and compared between the two groups. Results There were no significant differences between the group A and B with respect to the mean operative time [(84.0±24.6) min vs. (94.0±22.7) min, t=1.372, P=0.177], hospitalization time [(6.5±1.1)days vs. (6.3±1.8)days, t=0.49, P=0.261], blood transfusion quantity [(82.9±10.6) ml vs. (85.0±11.8) ml, t=0.415, P=0.682], kidney and colostomy channels [single channel(70.6% vs. 75.0%), double channel (29.4% vs. 25.0%), χ2 =0.22, P=0.638] and residual stones rate (5% vs. 3%,χ2=0.42, P=0.282). Conclusions When PCNL is performed after previous open nephrolithotomy, there is no difference in success rate and morbidities.

OBJECTIVETo better understand the molecular mechanism of tumorigenesis and progression, the monoclonal antibody against G3BP (Ras-GAP SH3 binding protein), which serves as an important downstream effector of Ras signaling, was prepared, characterized and utilized in analysis of various human tumors.

METHODSBy using the prokaryotic expression vector pGEX-5X1, GST-G3BP fusion protein was expressed in E. coli BL21 under induction of IPTG. Purified GST-G3BP fusion protein was used to immunize BALB/c mice. The monoclonal antibody against G3BP was produced through conventional hybridoma method and characterized by ELISA, Western blot and immunohistochemical staining.

RESULTSA hybridoma cell line secreting anti-G3BP IgG1 subtype antibody was obtained. Western blot and competitive inhibition assay showed that the antibody was G3BP-specific. Immunohistochemical staining demonstrated that G3BP was over-expressed in formalin-fixed and paraffin-embedded tissues of some human tumors, such as lung cancer, colon cancer, gastric cancer and breast cancer. In breast cancer specimens, the degree of G3BP expression correlated positively with the presence of lymph node metastasis and c-erbB2 expression.

CONCLUSIONSThe G3BP-specific monoclonal antibody derived from recombination protein can be used in ELISA, Western blot and immunohistochemical assay. It may provide an important tool in analysis of G3BP in in vitro and in vivo experiments. Besides, G3BP may serve as another prognostic marker for breast cancer.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Biomarkers, Tumor ; Breast Neoplasms ; metabolism ; pathology ; Carrier Proteins ; genetics ; immunology ; metabolism ; DNA Helicases ; Female ; Genetic Vectors ; Humans ; Hybridomas ; secretion ; Lymphatic Metastasis ; Male ; Mice ; Mice, Inbred BALB C ; Poly-ADP-Ribose Binding Proteins ; RNA Helicases ; RNA Recognition Motif Proteins ; Receptor, ErbB-2 ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Tumor Cells, Cultured

Recent evidence indicates that the aberrant neuronal expression of mitotic proteins in Alzheimer's disease (AD) brain may be related to AD pathological changes. To investigate whether the toxicity of beta-amyloid protein (Abeta) induces mitotic proteins expression in adult rat brain, we used immunohistochemical and integral optical density analytic method to analyze the adult rat brains, which had been injected with Abeta(25-35) into unilateral amygdala. Results showed that the levels of neurofibrillary tangle (NFT) related phosphorylated tau protein and apoptosis related protein Bax were increased in Abeta(25-35) injected rat brains, meanwhile the aberrantly expression of mitotic protein cyclin A and cyclin B1 was also detected at 7 d after operation, but the level of cyclin A decreased and cyclin B1 disappeared at 21 d. Immunofluorescence double labeling presented that cyclin B1 was partially co-localized with Bax or phosphorylated tau protein, whereas Bax and phosphorylated tau protein seldom co-localized. These results suggest that Abeta causes mitotic protein expression in adult brain neurons, which may die through apoptosis or may be affected by AD NFT-related tau phosphorylation.
Alzheimer Disease ; metabolism ; physiopathology ; Amygdala ; drug effects ; metabolism ; physiopathology ; Amyloid beta-Peptides ; toxicity ; Animals ; Cyclin A ; metabolism ; Cyclin B1 ; metabolism ; Male ; Neurons ; metabolism ; Peptide Fragments ; toxicity ; Phosphorylation ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism ; tau Proteins ; metabolism

OBJECTIVETo investigate the mechanism of influence of phosphoprotein associated with glycosphingolipid microdomains 1 (PAG1) on the biological behavior of human prostatic cancer cell line PC-3M-1E8.

METHODSThe expression of GST-Raf1-RBD recombinant protein, a specific binding domain of active Ras (GTP-Ras), was induced by IPTG in JM109 bacterium. SDS-PAGE and coomassie brilliant blue staining were performed using cleaved product of the bacterium to determine the expression of the fusion protein. GST-pull down essay was designed to detect the level of active Ras in PGA1 and vector transfected, respectively and in the native PC-3M-1E8 cells. Western blotting was used to detect the expression levels of downstream proteins of Ras signal pathway which may be related to the function of PAG1. Phalloidine labeled by tetramethylrhodamine-5-(and-6) isothiocyanate (TRITC) was used for the staining of intracellular F-actin, Laser passing confocal microscopy was adopted for observing change of the cell morphology and the arrangement of F-actin.

RESULTSAfter IPTG induction, the GST-Raf1-RBD recombinant protein, with a molecular weight of 33 000, was noticed to be highly expressed in JM109 bacterium. GST-pull down assay revealed that the expression level of active Ras markedly decreased after PAG1-transfection while the total Ras remained unchanged. The expression of p-ERK and cyclin D1 in the PAG1-transfected cells decreased in accordance with the level of active Ras. However, the expression of p21(WAF1/CIP1) and p-Akt didn't show any variation. Additionally, the structure of F-actin was turbulent and the pseudopodia of cells diminished conspicuously after PAG1-transfection. There was a high expression of PAG1 in normal human prostate tissue, however, the positive rate of PAG1 immuno-staining decreased in cases of prostatic adenocarcinoma, correspondent with increasing of the grading index of cell differentiation established in the Gleason grading system for the diagnosis of prostate adenocarcinoma.

CONCLUSIONSAn over-expression of PAG1 in PC-3M-1E8 cells effectively suppresses the activation of Ras and ERK, as well as the cyclin D1 expression, leading to an inhibition of the proliferation ability of tumor cells. The turbulence of F-actin and reduction of pseudopodia of cells result in an impairment of cell mobility, invasiveness and metastatic capability. In human prostate and prostatic adenocarcinoma tissues, the expression of PAG1 is related to the cellular differentiation and malignancy.

Actins ; metabolism ; Adaptor Proteins, Signal Transducing ; metabolism ; Adenocarcinoma ; metabolism ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Humans ; Male ; Membrane Proteins ; metabolism ; Neoplasm Invasiveness ; Prostate ; metabolism ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-akt ; metabolism ; Transfection ; ras Proteins ; metabolism

Purpose: To evaluate the use of signal intensity index (SII) of skull-base invasion in nasopharyngeal carcinoma (NPC) using magnetic resonance imaging (MRI), select a best cut-off SII value to predict the outcome of NPC. Materials and Methods: One hundred and twenty-two NPC patients (92 men, 30 women) with skull-base invasion were included. All patients underwent MRI, signal intensities on T1-weighted imaging (T1WI) were measured for each invaded site and its contralateral normal counterpart. The SIIs were calculated, receiver operating characteristic curves were constructed. The optimal cut-off values were extracted. The overall survival (OS) rates of 5-year follow-up were performed. Results: Sensitivities for differentiating skull-base invasion from normal contralateral anatomy were 98.9%, 88.5% and 70.0%, and specificities were 98.9%, 96.0% and 74.4%, respectively. There were three cut-off values for differentiating invasion from normal anatomy of skull-base, 49%, 98%, and 60%. Significant difference in OS rates (84.2% vs. 57.1%, p=0.007) was seen for SII threshold values > 60% and those ≤ 60%. Conclusions: The SII might be a useful means of differentiating invasion from normal tissue at the skull-base in NPC. The cut-off value of quantitative SII at the skull-base may aid in monitoring the response to treatment of NPC patients.

OBJECTIVETo investigate the regulatory mechanism of the transcription of tumor metastasis suppressor gene TMSG-1.

METHODSLuciferase reporter assay and site-directed mutagenesis were used to analyze the regulatory region of TMSG-1. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) were carried out to verify the interaction of KLF6 and Sp1 with the regulatory region of TMSG-1. Co-immunoprecipitation (CoIP) was performed to analyze the interaction between KLF6 and Sp1. TMSG-1 and wt-KLF6 mRNA expressions in cells with different metastatic capacities were quantitated by real-time PCR. Cell invasive capability was determined by Matrigel invasion assay.

RESULTSA 63 bp inducible regulatory region (+59 bp - +123 bp) in exon 1 was identified by luciferase assay using reporter plasmids with a series of TMSG-1 regulatory region deletions. Mutations in KLF6/Sp1 binding sites of this region resulted in a decrease of luciferase activity, while cotransfection with KLF6 or Sp1 expressing plasmids led to a remarkable increase of luciferase activity. EMSA and ChIP demonstrated that KLF6 as well as Sp1 interacted with this region. CoIP also indicated a possible interaction between KLF6 and Sp1 proteins. In the highly metastatic cell sublines, a low level of wild type KLF6 was associated synchronously with a low TMSG-1 level. Prostate carcinoma cells overexpressing KLF6 exhibited a higher TMSG-1 level and a lower invasive capability.

CONCLUSIONSTranscription factor complex of KLF6 and Sp1 may participate in the inducible transcriptional regulation of TMSG-1, and a decreased wild type KLF6 expression is likely associated with a low TMSG-1 level in the highly metastatic cell sublines.

Binding Sites ; genetics ; Cell Line, Tumor ; Electrophoretic Mobility Shift Assay ; Humans ; Immunoprecipitation ; Kruppel-Like Factor 6 ; Kruppel-Like Transcription Factors ; genetics ; metabolism ; Lung Neoplasms ; metabolism ; pathology ; Male ; Membrane Proteins ; genetics ; metabolism ; Mutagenesis, Site-Directed ; Mutation ; Neoplasm Invasiveness ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Sp1 Transcription Factor ; genetics ; metabolism ; Sphingosine N-Acyltransferase ; genetics ; metabolism ; Transcriptional Activation ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism

Objective To study the pharmacokinetics and safety in healthy volunteers following intravenous administration of biapenem.Methods Ten healthy volunteers were randomized to administer a single dose of biapenem at 300 and 600 mg,respectively.They were given the other dose after a week.The concentration of biapenem in human plasma and urine were determined by HPLC-UV.The main pharmacokinetic parameters were calculated with WinNonLin.Results The main pharmacokinetic parameters of biapenem after single doses of 300 and 600 mg were as follows C_(max) were(11.33±2.71)and(26.80±5.11)μg·mL~(-1);AUC_(0-tn) were(18.01±4.66)and(42.78±8.81)μg·mL~(-1)·h,respectively.Conclusion The process of biapenem in the dosage range of 300-600 mg fit linear dynamic feature.

Objective To investigate the genetic etiologies in the 0 -3 years old infants with hearing loss and to analyze the interaction between genetics and environmental factors. Methods Total of 130 infants were performed detailed audiological evaluation as well as the detection of the popular deafness gene mutations in GJB2 gene, SLC26A4 and mtDNAI2SrRNA. Of them, 84 cases were performed the computer tomography or magnetic resonance imaging examinations. Results Of the 130 cases, 54 infants were diagnosed as large vestibular aqueduct syndrome, while seven of 130 were as auditory neuropathy and the others were diagnosed as sensorineural hearing loss. Considering of the risks of etiologies for hearing loss, 85 of them had the experiences of the high risk factors at birth(65.4% ,85/130), while 23 of them had the exposure of aminoglycoside antibiotics, and 13 had the family history background as well as two eases were from the consanguineous families. In the causative genes screening, 42 infants were caused by the mutations of SLC26A4 gene (32.3%), but 14 infants found the mutations in GJB2 gene (4.6%), and no infants carried the mutation in mtDNA 12SrRNA 1555G and 1494T points in our studies. Conclusions In our studies, about 36. 9% infants hearing loss cases can be found the mutations in SLC26A4 and GJB2 genes. It is essential to put the idea into the hearing evaluation combined with genetic testing for the diagnoses of heating loss. It is also helpful for exploring the etiologies of hearing loss and performing the target genetic consulting for decreasing the prevalence of deafness in the future.

Objective To analyze the risk factors of cognitive impairment after off-pump coronary artery bypass grafting (OPCABG).Methods A total of 102 patients [ male:82,age:(65.7 ± 7.1 ) years ]undergoing OPCABG in our hospital between January 2009 and December 2010 were divided into postoperative cognitive dysfunction (POCD) group and non-POCD group by the MMSE questionnaire survey conducted at 7 days pre- and post-operation respectively.Results The incidence of POCD was 48.0％ ( 49/102 ).Multivariate logistic stepwise regression analysis showed:advanced age (OR =1.32,95％ CI:1.10 - 1.46,P=0.002),smoking (OR=1.26,95％ CI:1.18-1.32,P=0.001),hypertension (OR =1.66,95％ CI:1.36 - 1.78,P =0.023),diabetes ( OR =1.62,95％ CI:1.02 - 2.84,P =0.032),stroke ( OR =3.32,95 ％ CI:1.68 - 6.49,P ＜ 0.001 ),mitral regurgitation ( OR =1.48,95 ％ CI:1.26 - 1.89,P ＜ 0.001 ),and time of wall clamp ( OR =4.84,95％ CI:1.08 - 7.28,P ＜ 0.001 ) were independent risk factors of POCD.Conclusion Advanced age,smoking,hypertension,diabetes,stroke,mitral regurgitation,and prolonged time of wall clamp are major risk factors for POCD in patients undergoing OPCABG.

Objective:To compare the clinical efficacy of robot-assisted pedicle screw placement with visualization technology and conventional robot-assisted pedicle screw placement, and analyze the accuracy and safety of robot-assisted pedicle screw placement with visualization.Methods:This retrospective study analyzed data from 60 patients (39 males and 21 females) with an average age of 51.03±18.04 years (range 12-78 years) who underwent open spinal pedicle screw fixation surgery for thora columbar diseases at the Orthopedic Department of the First Affiliated Hospital of China Medical University between August 2020 and September 2022. The cases included 25 cases of spinal stenosis, 15 cases of lumbar fractures, 7 cases of thoracic fractures, 3 cases of lumbar spondylolisthesis, and 10 cases of spinal deformities. 30 patients underwent solid pedicle screw placement using robot-assisted visualization technology (visualization group), while the remaining 30 patients received hollow pedicle screw placement using conventional robot-assisted technology (conventional group). After screw placement, "O"-arm X-ray scans were performed for verification, and screw placement accuracy was evaluated based on the Gertzbein-Robbins standard. The study recorded and compared the time required for screw placement, number of fluoroscopy sessions, and perioperative complications between the two groups to provide a comprehensive assessment of surgical outcomes.Results:There were no significant differences in age and gender between the two groups ( P>0.05). In the visualization group, a total of 178 pedicle screws were placed, with 172 screws (96.6%) achieving satisfactory placement, while the conventional group placed 254 pedicle screws, with 240 screws (94.5%) achieving satisfactory placement. The difference in accuracy rates between the two groups was not statistically significant (χ 2=1.087, P=0.297). The visualization group required a mean of 2.60±1.03 fluoroscopy sessions during surgery, significantly less than the conventional group's mean of 5.57±2.12 sessions ( t=-6.860, P=0.001). Moreover, the visualization group had a shorter mean screw placement time of 13.23±3.68 minutes compared to the conventional group's mean of 24.68±15.75 minutes ( t=-3.870, P=0.040). All patients in both groups completed the surgery without postoperative complications such as infection, hematoma, or nerve root injury. Conclusion:The technique of robot-assisted pedicle screw placement with visualization effectively preserves the high precision achieved in conventional robotic surgery. With its advantage of real-time monitoring for screw position, it reduces the intraoperative fluoroscopy times and shortens the screw placement time, thereby further enhancing surgical efficiency.