Objective To investigate the iodide uptake by U251 glioma cell lines which were transfered with both human sodium/iodide symporter (hNIS) and human thyroperoxidase (hTPO) genes. Methods Recombinant adenosine virus AdTPO was constructed through cloning, recombination, packaging and amplifying. The viral titers were calculated after purification. The protein expression of AdTPO was tested by Western-Blotting and the recombinant plasmids PcDNA3. 1/hNIS were constructed. After hNIS gene was transfected into human glioma cell lines U251 through liposome, the cell lines with stable hNIS expression (hNIS-U251) selected by G418 antibiotics were defined as hNIS-U251 group. Then, hTPO was transducted into hNIS-U251 with adenosine virus (AdTPO-hNIS-U251 group). U251 cells with no plasmid were used as the control group (U251). Cultured cells from each group were studied for 125I uptake as well as 125I efflux rate. Student-Newman-Keuls in multiple range test was used. Results AdTPO-hNIS-U2.51 with stable expression was successfully established by transfecting hNIS and hTPO genes into human glioma cell lines. The 125I uptake by AdTPO-hNIS-U251, hNIS-U251 and U251 cell lines was (74 647.53 ±3605.88), (55 769.96 ±4353.26) and ( 507.67 ± 57.69 ) counts/min, respectively ( F = 836. 17, P ＜ 0.05 ). The uptake compacity by AdTPO-hNIS-U251 was 147 fold higher than that by U251 (q =55.64, P＜0.01 ) and 1.3 fold higher that by hNIS-U251 (q = 14. 17, P ＜0.01 ). 125I efflux rate was prolonged in AdTPO-hNIS-U251 group and its effective half time was 13 min. Conclusion Enhanced 125I uptake by the human glioma cell lines can be achieved with combined transfection of hNIS and hTPO genes.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; CD2 Antigens ; metabolism ; Child, Preschool ; Chromosome Breakage ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Ki-1 Antigen ; metabolism ; Lymphoma, Large-Cell, Anaplastic ; drug therapy ; metabolism ; pathology ; Mucin-1 ; metabolism ; Prednisone ; therapeutic use ; Receptor Protein-Tyrosine Kinases ; genetics ; metabolism ; Vincristine ; therapeutic use

This study aimed to investigate the involved immunologic mechanism of pure total flavonoids from Citrus (PTFC) on the development of non-alcoholic steatohepatitis (NASH). C57BL/6 mice were fed with high .fat diet for 16 weeks to induce the NASH model, and from the 7th week three dosages (25, 50 and 100 mg x kg(-1) x d(-1)) of PTFC were administrated intragastric for 10 weeks respectively. Serum TG, CHOL, ALT, AST were determined by biochemical assay, histopathological changes of the liver tissue were observed by HE staining, expression of RORyt and Foxp3 mRNA of the liver tissue was detected by Real-time PCR, and serum IL-17, IL-6, IL-10 and IL-4 were determined by.Cytometric Beads Array. As a result, we find that after the administration of PTFC, the in- flammation of the liver tissue of NASH mice was attenuated, liver function was improved, and the expression of RORgammat mRNA was higher in the liver tissue while which was lower of Foxp3 mRNA, the level of proinflammatory cytokines IL-17 and IL-6 decreased and the level of suppressive cytokines IL-10 and IL-4 increased. These data show that PTFC protects the development of NASH through regulating the Th17/Treg balance and attenuating inflammation.
Animals ; Citrus ; chemistry ; Cytokines ; blood ; Flavonoids ; pharmacology ; Male ; Mice ; Mice, Inbred C57BL ; Non-alcoholic Fatty Liver Disease ; immunology ; prevention & control ; T-Lymphocytes, Regulatory ; drug effects ; Th17 Cells ; drug effects

OBJECTIVETo observe the effect of pure total flavonoids from Citrus (PTFC) on the hepatic fatty degeneration, inflammation, oxidative stress and SIRT1/PGC-1alpha expressions in mice with non-alcohol steatohepatitis (NASH), and discuss the action mechanism of PTFC on NASH.

METHODMice were given high-fat diet for 16 weeks to induce the NASH model. Since the seventh week after the model establishment, the mice were intervened with 100, 50 and 25 mg x kg(-1) x d(-1) PTFC for 10 weeks. The pathologic changes in hepatic tissues were observed with HE staining. The contents of TG, CHOL in hepatic tissue, as well as the levels of AST, ALT in serum were detected by using the biochemical process. The expression of SIRT1, PGC-1alpha and MnSOD mRNA in hepatic tissues were detected with Real-time PCR assay. SIRT1, PGC-1alpha protein and 8-OHdG expressions were determined with the immunohistochemical method. The SOD level in hepatic tissues was tested by the xanthine oxidase method. The MDA content in hepatic tissues was examined by the thiobarbituric acid method.

RESULTThe contents of TG, CHOL, NAFLD activity scores and ALT level in serum in hepatic tissues of mice in the model induced by fat-rich diet were obviously higher than that of the normal group (P < 0.010. The SIRT1, PGC-1alpha, MnSOD mRNA and protein expression in hepatic tissues were significantly lower than that of the normal group (P < 0.01). The expression of 8-OHdG and the content of MDA in hepatic tissues were obviously higher than that of the normal group (P < 0.01). After the intervention with different doses of PTFC, the NAFLD activity scores, the content of TG and the level of AST in serum were notably lower than that of the normal group (P < 0.01, P < 0.05); whereas the SIRT1, PGC-1alpha, MnSOD mRNA and protein expression were obviously higher than that of the normal group (P < 0.01, P < 0.05), with the significant decrease in the expression of 8-OHdG and the content of MDA (P < 0.01).

CONCLUSIONOxidative stress/lipid peroxidation enhancement in in NASH mice induced by high-fat diet may be related to the changes in SIRT1/PGC-1alpha signal transduction pathway. PTFC could enhance the anti-oxidant capacity in liver, relieve the damage of reactive oxygen during the fatty acid metabolic process, and prevent NASH from the occurrence and development by regulating the SIRT1/PGC-1alpha signal pathway.

Animals ; Citrus ; chemistry ; Fatty Liver ; drug therapy ; genetics ; metabolism ; Flavonoids ; chemistry ; pharmacology ; Inflammation ; drug therapy ; genetics ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Non-alcoholic Fatty Liver Disease ; Oxidative Stress ; drug effects ; genetics ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; Sirtuin 1 ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism

Objective To investigate the distribution and antibiotic resistance of bacterial isolates from blood samples in pediatric patients in tertiary hospitals in Hubei area from 2006 to 2007.Methods Pathogenic bacteria isolated from blood samples of pediatric patients were collected from 17 tertiary hospitals in Hubei area from 2006 to 2007. All strains were isolated and identified by routine Methods . Antimicrobial susceptibility testing was conducted on all isolates using Kirby-Bauer Methods . Results A total of 941 strains were collected from January to December of 2006. The most common microorganism was coagulase-negative Staphylococcus (573, 60.9%), followed by Staphylococcus aureus (127, 13.5%), Enterococcus faecalis (33, 3.5%), Escherichia coli (16, 1.7%). A total of 969 strains were collected from January to December of 2007. The most common species was coagulase negative Staphylococcus (583, 60.2%), followed by S. aureus (162, 16.7%), E. faecalis (28, 2.9%), E. coli (21, 2.2%), E. faecium (11, 1.1%), Salmonella choleraesuis (11, 1.1%). Of the isolates collected during 2006, the prevalence of methicillin-resistant S. aureus (MRSA) was 71.7%. The prevalence of ESBLs was 56.2% in E. coli. Of the isolates collected during 2007, the prevalence of MRSA was 79.6%. The prevalence of ESBLs was 47.6% in E. coli. MRSA strains were more resistant to antibiotics than methicillin-susceptible S. aureus (MSSA). No glycopeptide-resistant strain was identified in Staphylococcus. Conclusions Staphylococcus is the most frequently isolated pathogen from blood samples of pediatric patients in tertiary hospitals in Hubei area.

Objective To investigate distribution and drug resistance of pathogenic bacteria in lower respitatory tract infection. Methods Distribution and drug resistance of pathogenic bacteria in lower respitatory tract infection of patients in ICU and non-ICU of our hospital during 2013 were retrospectivly analyzed. The pathogens were identified by manual methods routinely and those difficult to be identified were analyzed by using the VITEK-2-COMPACT instrument. Antimicrobial susceptibility of these isolates were tested by Kirby-Bauey methods routinely. Results In total, 956 strains were isolated from lower respitatory tract infection of patients in ICU, including 231 strains of gram-positive cocci (24. 2%), 680 strains of gram-negative bacteria (71. 1%), 45 strains of fungi (4. 7%). In patients of non-ICU, 4 464 strains were isolated, including 1 090 strains of gram-positive cocci (24. 4%), 3 226 strains of gram-negative bacteria (72. 3%), and 148 strains of fungi (3. 3%). Staphylococcus aureus, acinetobacter baumannii and pseudomonas aeruginosa were the most frequent isolates in patients of ICU and non-ICU. The overall prevalence of methicillin resistant staphylococcus aureus (MRSA) in staphylococcus aureus was 87. 0%in ICU and 74. 0% in non-ICU. MSSA was sensitive to the most antibiotics ( more than 80. 0% of the strains were sensitive to common antibiotics) except penicillin, erythromycin and clindamycin. MRSA was sensitive to trimethoprim-sulfamethoxazole and fosfomycin (more than 75. 0% of the strains were sensitive to the antibiotics) except for vancomycin, teicoplanin and linezolid. Acinetobacter baumannii was more resistant to the antibiotics (less than 40. 0% of the strains were susceptible to the antibiotics). Pseudomonas aeruginosa from ICU was more resistant to the antibiotics ( less than 50. 0% of the strains were sensitive to the antibiotics) than that from non-ICU. Stenotrophomonas maltophilia was sensitive to trimethoprim-sulfamethoxazole, levofloxacin and minocycline (more than 80. 0% of the strains were sensitive to the antibiotics). Escherichia coli and Klebsiella pneumoniae were sensitive to Piperacillin-tazobactam and Amikacin except for meropenem and imipenem ( more than 80. 0% of the strains were sensitive to the antibiotics) . Conclusion Gram-negative bacteria was the most frequent isolate in lower respitatory tract infection of our hospital during 2013. Staphylococcus aureus, acinetobacter baumannii and pseudomonas aeruginosa were the most frequent isolates in ICU and non-ICU. Resistance to the antibiotics was more common in ICU than in non-ICU. Antibiotics should be prescribed according to bacterial resistance results reasonably in order to prevent the spread of drug-resistant strains.

Objective To investigate the antimicrobial resistance in the clinical strains isolated from Tongji Hospital to the antimicrobial agents commonly used in 2012.Methods Antimicrobial susceptibility was tested by Kirby-Bauer method.The minimum inhibitory concentration (MIC) of penicillin and ceftriaxone for Streptococcus pneumoniae and vancomycin for Staphylococcus spp.were determined by E-test.All data were analyzed by WHONET 5.6 software.Results A total of 8 191 strains were isolated in 2012,including gram-positive bacteria (2 815,34.4%)and gram-negative bacteria (5 376,65.6%). The top five pathogenic bacteria isolated from outpatients were Escheria coli,Pseudomonas aeruginosa,coagulase-negative Staphylococcus (CNS),Klebsiella spp.and Staphylococcus aureus.For the isolates from non-ICU inpatients,the top five were E.coli,S.aureus,Acinetobacter spp.,Klebsiella spp.and P.aeruginosa.For those isolated from ICU patients,the top five were Acinetobacter spp.,S.aureus,P.aeruginosa,Enterococcus spp.and E.coli.The prevalence of MRSA and MRCNS was 58.1% and 64.3%,respectively.Seventeen strains of vancomycin resistant Enterococcus were identified, including 13 strains of E.faecium with VanA and 4 strains of E.gallinarum with VanA and VanC.The percentage of antimicrobial resistance in E.faecium was significantly higher than that in E.faecalis (P<0.05).A total of 94 strains of carbapenem-resistant Enterobacteriaceae were detected.The prevalence of penicillin-non-susceptible S.pneumoniae in children was much higher than that in adults. The prevalence of carbapenem-resistant P.aerugonosa and Acinetobacter spp. was 28.1% and 56.2% respectively.Beta-lactamase was produced in 41.8% of the H.influenzae and 98.6% of the M. catarrhalis isolates. Conclusions The prevalence of multidrug resistant strains has been increasing, especially vancomycin-resistant Enterococcus and carbapenem-resistant Enterobacteriaceae.

OBJECTIVETo introduce an arthroscopic technique in managing recurrent dislocation of the patella and its clinical results.

METHODSSixteen patients with recurrent patellar dislocation were reviewed, including 3 males and 13 females. The average age was 17.6 years old (ranged from 14 to 32 years). The patients suffering from patellar sub-luxation averaged 18.5 months (ranged from 6 to 23 months)before operation. These patients were treated with lateral patellar retinacular release outside the synovial bursa of knee joint and medial patellofemoral ligament reconstruction using the semitendinosus tendon free autograft. The Lyshohm scores before and after operation were used to evaluate outcomes at the final follow-up.

RESULTSAll the patients were followed up, and the duration ranged from 6 to 48 months, with an average of 12 months. There was no recurrence. The Q angle decreased from (16.4 ± 3.7)° to (10.1 ± 1.4)°; insall index decreased from 1.37 ± 0.25 to 1.28 ± 0.23; congruence angle decreased from (21.3 ± 2.6)° to (5.86 ± 2.23)°; Lysholm score improved from 76.1 ± 5.2 to 89.8 ± 4.1 at 6 months after operation.

CONCLUSIONCompared with conventional procedure, arthroscopic surgery for recurrent dislocation of the patella achieves excellent outcomes with minimum invasion.

Adolescent ; Adult ; Arthroscopy ; Bursa, Synovial ; surgery ; Female ; Humans ; Knee Joint ; surgery ; Male ; Patellar Dislocation ; physiopathology ; surgery ; Patellar Ligament ; surgery ; Range of Motion, Articular ; Treatment Outcome ; Young Adult

Objective To evaluate the effect of hyperhomocysteinemia on ischemic diease in rats. Methods Thirty-two SD rats were randomized into two groups,control group(n=16,administered with tap water) and hyperhomocysteinemia group(HH group,n=16,administered with water containing L-methionine at 1 g/kg/d).At the 14thday of dietary modification,the left femoral artery and vein were excised,and the interventions continued for another 14 days.At the 15th day after operation,serum biochemical parameters as well as NOx and cGMP in ischemic tissues were tested,capillary vessel density of both hindlimbs were measured by histological analysis,and angiogenesis of ischemic hindlimb was observed by angiography. Results At the 15th day after operation,the level of high-density lipoprotein-cholesterol(HDL-C),blood urea nitrogen,uric acid and ratio of blood urea nitrogen to serum creatinine were significantly higher in HH group than those in control group,while the level of serum folic acid,VitB12,HDL-C and endogenous creatinine clearance rate were significantly lower in HH group than those in control group(P

Objective To study the inhibitory effect of pterin acid (PTA) against ricin and recombinant ricin A chain protein. Methods Luciferase protein synthesis inhibition assay in a cell-free system and in vitro cytotoxicity experiments were performed to assess the biological activity of ricin and rRTA treated with PTA.Results The result showed that PTA could significantly inhibit the activity of ricin and rRTA in a dose-dependent manner.Conclusion PTA might be used as a small molecular probe to develop an evaluating system for ricin/RTA small molecular inhibitor in vitro. The cell-free system adopted in the current study could also serve as a necessary basis for screening some novel small molecular compounds against ricin and RTA in the future.