Background Toll-like receptor 4 (TLR4) is an immune related receptor.It plays an important role in inducing inflammation response.The inflammatory response secondary to optic nerve crush will results in serious retinal damage.It is worthy of studying the expression and effect of TLR4 in retina after optic nerve crush.Objective This experimental study was to explore the role of TLR4 in the loss of retinal ganglion cells(RGCs) after optic nerve crush.Methods Twenty-four SPF adult health Sprague-Dawley (SD) rats were used in the study and radomized into two groups based to the experimental time.The optical nerve crush models were established by crushing the optical nerve in the right eyes of the rats,and the left normal eyes served as controls.The rats were sacrificed by over anesthesia and retinas were isolated 3 days and 7 days after operation.Expression of TLR4 in the retinas was detected using immunofluorescence method.Reverse trancription PCR (RT-PCR) and Western blot were applied respectively for the detection of TLR4 mRNA and protein in the retinas.The apoptosis of RGCs was evaluated using TUNEL staining.The use and care of experimental animals followed theGuide for the Care and Use of Laboratory Animals of NIH.This study was approved by the Institutional Animal Care and Use Committee at the Zhongshan Ophthalmic Center.Results The expression of TLR4 in rat retinas presented with green fluorescence mainly in the inner layer of retinas.The fluorescence was enhanced in the model 3 days group and the model 7 days group compared with the corresponding control groups.The relative expressing values of TLR4 mRNA in the retinas were 2.92±0.06and 3.92±0.12 in the model 3 days and 7 days groups,respectively,which were significantly higher than 2.87±0.12and 3.44±0.17 in the control 3 days and 7 days group (t3d =-12.888,P＜0.001 ;t7d =-4.669,P=0.010).In the model 3 days group and model 7 days group,the grey values of TLR4 protein were 1.14±0.05 and 1.49±0.03,and those in the control 3 days and 7 days groups were 0.99 ± 0.09 and 1.38 ± 0.07,showing significant differences between them(t3d =-11.324,P＜0.001 ;t7d =-5.638,P=0.005).Apoptotic RGCs were obviously increased in the optic nerve damage group in comparison with the control group.Conclusions The TLR4 is over-expressed in the inner layer of retina after optic nerve crush,which suggestes that TLR4 is probably involved in the loss of RGCs after optic nerve crush.

Antineoplastic Agents ; therapeutic use ; Apoptosis ; drug effects ; Caspases ; metabolism ; physiology ; Enzyme Activation ; Fas Ligand Protein ; metabolism ; Humans ; NF-kappa B ; metabolism ; Neoplasms ; metabolism ; therapy ; Neovascularization, Pathologic ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Signal Transduction

Objective To explore the pathophysiological mechanisim underlying the development and progression of chronic subdural hematoma (CSDH) through an ultrastructural observation of the outer membrane of CSDH. Methods A total of 8 samples of CSDH outer membrane were obtained from the 8 patients who had uncergone surgery of hematoma removal in our department from January 2008 to January 2009.CT scanning revealed 2 cases of low hematoma density,2 cases of hematoma isodensity, 2 cases of high hematoma density and 2 cases of mixed hematoma desity.Conventional light microscopy and electron microscopy were used to observe the ultrastructure of the outer membrane. Results Light microscopy showed numerous dilatated and congested macrocapillaries with a wide vascular lumen in the outer membrane of the hematoma capsule.Electron microscopy showed weak, discontinous or partially dissolved endothelial cells in the macrocapillaries.Scattered red blood cells in the extracellular space were found, indicating bleeding within the outer membrane.Eosinophils increased with enlarged granules within the cellular cytoplasm.Neutrophils and macrophages were also present in some specimens.Fibroblasts showed a state of significant proliferation and activation. Conclusions There are abundant newly formed vascular networks in the outer membrane of CSDH. Neomembrane formation, neovascularization and repeated micro-haemorrhages from these fragile new vessels may play a key role in the development and progression of CSDH.

Objective To investigate in vitro activity of antimicrobial agents against Enterococcus spp . isolated from clinic specimens in a hospital.Methods 188 Enterococcus spp . isolates from specimens sent by clinic depart-ments in June 2013-July 2014 were identified and performed antimicrobial susceptibility testing.Results Of 188 En-terococcus spp . isolates,119 were Enterococcus faecium (E.faecium),60 were E.faecalis ,and 9 were E.avium, these strains were mainly isolated from urine (34.57%)and blood specimens (19.15% ).No daptomycin and linezolid-resistant strain was detected;resistant rates of E.faecium to vancomycin was 1 .68%,to penicillin, ampicillin,high concentration gentamycin,erythromycin,and levofloxacin were all > 70%;except tetracycline, resistant rates of E.faecalis to the other antimicrobial agents were all lower than E.faecium,resistant rates of E. faecalis to penicillin and ampicillin were 16.67% and 13.33% respectively.Conclusion Daptomycin has high activity against Enterococcus spp . in this hospital.

OBJECTIVE: To investigate genetic polymorphisms of 21 non-CODIS STR loci in Han population from the east of China and to explore their forensic application value. METHODS: Twenty-one non-CODIS STR loci, were amplified with AGCU 21+1 STR kit and DNA samples were obtained from 225 unrelated individuals of the Han population from the east of China. The PCR products were analyzed with 3130 Genetic Analyzer and genotyped with GeneMapper ID v3.2 software. The genetic data were statistically analyzed with PowerStats v12.xls and Cervus 2.0 software. RESULTS: The distributions of 21 non-CODIS STR loci satisfied the Hardy-Weinberg equilibration. The heterozygosity (H) distributions were 0.596-0.804, the discrimination power (DP) were 0.764-0.948, the probability of exclusion of duo-testing (PEduo) were 0.176-0.492, the probability of exclusion of trios-testing (PEtrio) were 0.334-0.663, and the polymorphic information content (PIC) were 0.522-0.807. The cumulative probability of exclusion (CPE) of duo-testing was 0.999707, the CPE of trios-testing was 0.9999994, and the cumulated discrimination power (CDP) was 0.99999999999999999994. CONCLUSION Twenty-one non-CODIS STR loci are highly polymorphic. They can be effectively used in personal identification and paternity testing in trios cases. They can also be used as supplement in the difficult cases of diad paternity testing.
Alleles ; Asian People/genetics* ; China/ethnology* ; DNA/isolation & purification* ; Forensic Genetics/methods* ; Gene Frequency ; Genetic Loci/genetics* ; Genotype ; Humans ; Microsatellite Repeats/genetics* ; Polymerase Chain Reaction ; Polymorphism, Genetic

Objective To detect the feasibility of magnetically labeled swine bone marrow mesenehymal stem cells(MSCs)with SHU 555A combined with poly-L-arginine(PLL),under MR imaging in vitro and in vivo.Methods Swine mesenehymal stem cells were isolated and culture-expanded 3 passages in vitro,then magnetically labeled by incubation with SHU 555A(25?g Fe/ml,Resovist,Schering)for 24 hours with 750 ng/mL poly-L-lysine(PLL;average MW_275 kDa)added 1 hour before incubation.Cellular iron incorporation and detention at 0 d,4 d,8 d,12 d,16 d,20 d after labeling was qualitatively assessed using Prussian blue and quantified at atomic absorption spectrometry.Cell viability was assessed by trypan-blue exclusion test.Cell suspensions underwent MR imaging with T_1-and T_2-weighted spin-echo and fast field-echo sequences on a clinical 1.5 T MR system.At last,1?10~6 SHU 555A labeled and unlabeled MSCs were transextracardially implanted into the infracted and normal myocardium approximately 2 week following the ligation of left anterior descending coronary artery in 1 swine respectively,and finally performed 1.5-T MRI within 1 week after infarction.Results①Intracytoplasmic particles stained with Prussian blue stain were detected for all cells with mean cellular iron content of(13.13?2.30)pg per cell.With division of stem cells, the stained particles decreased gradually with iron content(0.68?0.20)pg per cell.at 16 days after labeling, approximately to the prelabeled baseline values.(0.21?0.06)pg per cell(P＞0.05).The viability of the labeled cells at various time points were not significantly different with that of nonlabeled cells(P＞0.05).②MR images showed signal intensity changed most obviouly in T2*WI in vitro.The percentage change of signal intensity increased with increasing cell numbers,and decreased with the time.As few as 5?10~4-1?10~5 cells could be detected by using this approach.③Two injected sites containing MR-MSCs were detected in vivo,presentingas low signal intensity areas with the T_2*WI scanning sequence.Conclusion Swine bone marrow MSCs can be labeled with SHU555A-PLL and depicted with a standard 1.5-T MR imager in vitro and in vivo.(J lntervent Radiol,2007,16:115-121)

OBJECTIVETo find out the optimal approach to decompress externally the severe injured brain and to avoid possible complications caused by external decompression.

METHODS68 patients who underwent external decompression after traumatic brain injury were admitted into Tianjin Medical University General Hospital for cranioplasty from 1995 to 2001. Complications were retrospectively investigated and analyzed in all patients. The findings were compared between the patients who accepted the decompressive craniectomy in our hospital and in local hospitals. chi(2)-test was employed for statistical analysis and complication evaluation.

RESULTSLarge craniectomy definitely caused some side effects to patients. Among various complications, several of them showed significantly high incidence (P<0.05) in patients who underwent the decompressive operation in local hospitals such as shunt-dependent hydrocephalous, subdural fluid collection, and CSF leakage from scalp incision. The rest of the complications had no remarkable difference (P<0.05) between the two groups including dilation or/and migration of lateral ventricle underlying the cranial defect, skin flap concavity, encephalomalacia of the decompressive area, seizure and infection.

CONCLUSIONSTo reduce the incidence of iatrogenic side effects, surgical craniectomy should be performed according to the strict indication and standard and any abuse should be avoided.

Adolescent ; Adult ; Chi-Square Distribution ; Craniocerebral Trauma ; surgery ; Craniotomy ; adverse effects ; standards ; Decompression, Surgical ; adverse effects ; standards ; Female ; Humans ; Male ; Middle Aged ; Postoperative Complications ; epidemiology ; Treatment Outcome

OBJECTIVETo establish a high-throughput chemiluminescence assay of serotype 5 specific neutralizing antibody and understand the epidemiology of this antibody in the healthy adults and children in Guangzhou.

METHODSUsing rAd5 carrying the reporter gene of secreted alkaline phosphatases (SEAP), serum samples from 116 healthy adults and 94 healthy children were examined with chemiluminescence assay to detect the presence of Ad5 neutralizing antibody. The reliability of this assay was tested against conventional cytopathic effect observation.

RESULTSThe chemiluminescence assay using secreted alkaline phosphatases (SEAP) as the reporter allowed rapid, sensitive, specific and reproducible detection of serotype 5 specific neutralizing antibody for epidemiological study of Ad5, which was positive in 26.72% of the adults and 17.02% of the children in this study.

CONCLUSIONAd5 neutralizing antibody is prevalent in the population of Guangzhou, suggesting the necessity of developing other serotype adenovectors for better vaccination and therapeutic effects.

Adenoviridae ; classification ; genetics ; immunology ; Adult ; Alkaline Phosphatase ; genetics ; Animals ; Antibodies, Neutralizing ; immunology ; Artifacts ; Blotting, Western ; China ; DNA, Recombinant ; genetics ; Female ; Genes, Reporter ; genetics ; High-Throughput Screening Assays ; Humans ; Luminescent Measurements ; methods ; Male ; Middle Aged ; Reproducibility of Results ; Serotyping ; Time Factors ; Young Adult

This study is to investigate the antitumor activities of the immunoconjugates composed of anti-type IV collagenase monoclonal antibody 3G11 and lidamycin (LDM) prepared by different methods. The immunoconjugates were prepared by linking 2-iminothiolane modified 3G11 to lysine-69 of LDM apoprotein by SPDP and SMBS as the intermediate drug linker. Immunoreactivity of the conjugates was determined by ELISA. The cytotoxicity of the conjugates was examined by clonogenic assay. Antitumor effects of the conjugates in vivo were evaluated in nude mice bearing subcutaneously implanted HT-1080 tumor. ELISA assay showed that the immunoconjugates retained the immunoreactivity of 3G11 against type IV collagenase. The cytotoxicity of the 3G11-SMBS-LDM to HT-1080 cells was significantly more potent than that of free LDM and 3G11-SPDP-LDM. In animal model at the same condition, free LDM inhibited the growth of HT-1080 tumor by 71.2%, while 3G11-SPDP-LDM and 3Gl1-SMBS-LDM reached 77.1% and 86.1%, respectively. The median survival time of the mice treated with free LDM was prolonged by 71.9% compared with that of untreated group. Whereas, the median survival time of 3G11-SPDP-LDM and 3G11-SMBS-LDM was prolonged by 125.3% and 163.7%, respectively, indicating that 3G11-SMBS-LDM was more effective than 3G11-SPDP-LDM in tumor suppression and life span prolongation. 3Gll-SMBS-LDM has more selective antitumor efficacy and lower toxicity, and might be a novel candidate for cancer therapy. LDM was more effective than 3G11-SPDP-LDM in tumor suppression and life span prolongation. 3Gll-SMBS-LDM has more selective antitumor efficacy and lower toxicity, and might be a novel candidate for cancer therapy.
Aminoglycosides ; therapeutic use ; Animals ; Antibiotics, Antineoplastic ; therapeutic use ; Antibodies, Monoclonal ; immunology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Collagenases ; immunology ; Enediynes ; therapeutic use ; Fibrosarcoma ; pathology ; therapy ; Humans ; Immunoconjugates ; therapeutic use ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Tumor Burden ; drug effects

OBJECTIVETo evaluate the efficacy and safety of recombinant adenovirus-p53 gene (Gendicine) therapy combined with radiotherapy for head and neck squamous-cell carcinoma (HNSCC).

METHODSFrom Oct. 2001 to May 2003, a randomized controlled clinical trial on Gendicine combined with radiation in 36 patients (gene therapy + radiotherapy, GTRT) vs. radiotherapy alone in 33 patients (RT) with HNSCC was completed. In the GTRT group, Gendicine 1 x 10(12) VP (virus particle) was injected intratumorally once a week for eight weeks, and concurrently followed by irradiation. For both groups, the conventional fractionation 2 Gy/f, five fractions a week to a total dose of 70 Gy, was given to either primary tumor or neck lymph nodes. Tumor response was assessed by CT image at 40 Gy, 70 Gy, 2 months after treatment to evaluate the response rate of CR, PR, SD and PD.

RESULTSWild-type p53 gene significantly enhanced radiotherapeutic effectiveness in patients with HNSCC (P < 0.05). The CR rate of tumors treated by GTRT was increased by nearly 2.31 times more than that of tumors treated by RT alone. No dose-limiting toxicity and adverse events were noted, except transient fever after Gendicine administration.

CONCLUSIONIntratumoral injection of Gendicine to HNSCC patients is safe and effective. The apparent improved results of combined therapy with Gendicine and radiation suggest that p53 gene therapy has promising therapeutic potential in cancer treatment.

Adenoviridae ; genetics ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; radiotherapy ; therapy ; Combined Modality Therapy ; Female ; Genes, p53 ; Genetic Therapy ; methods ; Genetic Vectors ; administration & dosage ; genetics ; Head and Neck Neoplasms ; radiotherapy ; therapy ; Humans ; Injections, Intralesional ; Male ; Middle Aged ; Recombinant Fusion Proteins ; administration & dosage ; genetics