Background Age-related cataract is one of the common causes of blindness.Although the pathophysiology of age-related cataract is far from clearly understood,it is well accepted that DNA damage plays an important role in the disease pathogenesis.Objective The purpose of this study was to quantitatively evaluate the DNA damage in peripheral lymphocytes of age-related cataract.Methods A cross-sectional study was carried out.This study complied Declaration of Helsinki and approved by Ethic Committee of Affiliated Hospital of Nantong University.Written informed consent was obtained from each subject.Two hundred and eleven patients with agerelated cataract and 147 normal subjects were enrolled from a “ Jiangsu Eye Study:Funing 2011 Eye Disease Epidemic Survey”.All the subjects aged from 50 through 80 years with matched age and gender between the two groups.The percentage of tail DNA and Olive tail moment (OTM) were detected by comet assay to assess the extent of DNA damage in peripheral lymphocytes.Statistical analyses were performed with SPSS 17.0 software,and the differences of the percentage of tail DNA and OTM were compared between the age-related cataract group and normal control group by independent sample t test as well as among the 50-59 years group,60-69 years group and ≥70 years group by one-way analysis of variance.Results Comet assay showed a round lymph cell with the clear border in the normal group;while in the age-related cataract group,the cell was bigger with a comet-like tail.The percentage of tail DNA and OTM in peripheral lymphocytes were (21.75 ± 3.51) ％ and 6.54 ± 1.65 in the age-related cataract group,and those in the normal control group were (9.31 ±3.60)％ and 2.18 ± 1.10,respectively,with significant differences between them (t =32.67,P =0.00 ; t =28.02,P =O.00).In the 50-59 years subgroup of the age-related cataract group,the percentage of tail DNA and OTM in peripheral lymphocytes were (20.04±2.86) ％ and 5.92± 1.14,and in the 60-69 years subgroup of the age-related cataract group,the percentage of tail DNA and OTM in peripheral lymphocytes were (20.77 ±2.93) ％ and 6.13 ± 1.14,which were significantly reduced in comparison with (22.79 ± 3.67)％ and 6.95±1.91 of the ≥70years subgroup(TailDNA％:q=2.75,P=0.00; q=2.02,P=0.00;OTM:q=1.03,P =0.02 ; q =0.82,P =0.00).Conclusions The pathogenesis and development of age-related cataract probably is associated with DNA damage.

Adult ; Female ; Hepatitis B, Chronic ; metabolism ; Humans ; Liver ; metabolism ; Male ; Receptors, CCR5 ; biosynthesis ; genetics ; Receptors, CXCR3 ; Receptors, Chemokine ; biosynthesis ; genetics

OBJECTIVETo investigate the expression of thymosin beta10 (Tbeta10) and related changes of actin filament organization in human tumor cell lines with different metastatic potential.

METHODSFour groups of nine human tumor cell lines with different metastatic potential were analyzed for the expression of Tbeta10 mRNA detected by northern-blot and its peptide by immunohistochemical staining. The filamentous actin (F-actin) was stained with TRITC-phalloidin to detect changes in actin organization.

RESULTSIn comparison with the non and/or weakly metastatic counterparts, Tbeta10 was upregulated in highly metastatic human lung cancer, malignant melanoma and breast cancer cell lines. TRITC-phalloidin staining revealed less actin bundles and a fuzzy network of shorter filaments in the highly metastatic tumor cells, while in the non and/or weakly metastatic cancer cell lines, there were thick and orderly arranged actin filaments.

CONCLUSIONSTbeta10 levels correlate positively with the metastatic phenotype in human tumors currently examined. The increased metastatic potential of tumor cells is accompanied by the loss of F-actin and poorly organized actin skeleton. There is a consistent correlation between the elevated Tbeta10 expression and the disrupted actin skeleton.

Actins ; analysis ; Blotting, Northern ; Cell Line, Tumor ; Humans ; Immunohistochemistry ; Neoplasm Metastasis ; Thymosin ; analysis

BACKGROUNDTo investigate the differential expression levels of thymosin beta 10 (T beta 10) and the corresponding changes of actin filament organization in human tumor cell lines with different metastatic potential.

METHODSFour groups of nine human tumor cell lines with different metastatic potential were analyzed for the amount of T beta 10 mRNAs by Northern blot and for their peptide expression levels by immunohistochemistry. The filamentous actin (F-actin) was observed by staining of TRITC-phalloidin to detect changes in actin organization.

RESULTSIn comparison with non-/weakly metastatic counterparts, T beta 10 was upregulated in highly metastatic human lung cancer, malignant melanoma and breast cancer cell lines. Staining of TRITC-phalloidin revealed less actin bundles, a fuzzy network of shorter filaments and some F-actin aggregates in the highly metastatic tumor cells. Meanwhile, the actin filaments were robust and orderly arranged in the non-/weakly metastatic cancer cell lines.

CONCLUSIONT beta 10 levels correlate positively with the metastatic capacity in human tumors currently examined. The increasing metastatic potential of tumor cells is accompanied by a loss of F-actin, poorly arranged actin skeleton organizations and presence of F-actin aggregates. There is a consistent correlation between the elevated T beta 10 expression and the disrupted actin skeleton.

Actin Cytoskeleton ; ultrastructure ; Blotting, Northern ; Cell Line, Tumor ; Humans ; Immunohistochemistry ; Neoplasm Metastasis ; genetics ; ultrastructure ; RNA, Messenger ; analysis ; Thymosin ; analysis

OBJECTIVETo investigate neuron regeneration of aged rats after intracerebral hemorrhage(ICH).

METHODSICH model was induced by intracerebral infusion of 100microL autologous blood in adult and aged (3 and 18 m of age, respectively) male Sprague-Dawley (SD) rats. Doublecortin(DCX), polysialylated neural cell adhesion protein(PSA-NCAM) and Bromodeoxyuridine (Brdu) in brain tissue were detected by Western blot analysis and immunohistochemical examination.

RESULTIn aged rats, DCX levels in the ipsilateral basal ganglia and subventricular zone started to increase at 7d after ICH, peaked at 14 d [(670+/-145)% of controls, P<0.01]. Most DCX positive cells in ipsilateral striatum were BrdU positive. Though DCX content in ipsilateral basal ganglia [(2560+/-758) pixels]of ICH aged rats was much higher than that of aged rat controls [(450+/-97)pixels, P<0.01], it was still lower than that in adult rats.

CONCLUSIONThe results demonstrate that neuron regeneration occurs in subventricular zone in the aged rats following intracerebral hemorrhage(ICH).

Age Factors ; Animals ; Cerebral Hemorrhage ; physiopathology ; Male ; Nerve Regeneration ; physiology ; Neuronal Plasticity ; physiology ; Neurons ; physiology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate how F261S mutation identified from Chinese obese patients affects the function of melanocortin 4 receptor (MC4R) and to analyze the obesity-related phenotypes in subjects carrying the F261S mutation.

METHODSF261S mutant of MC4R was generated by site-directed mutagenesis. Plasmids encoding wild-type or F261S mutant of MC4R were transfected into HEK293 and COS-7 cells to examine their functional characteristics. Signaling properties of F261S MC4R were assessed by measuring intracellular cAMP levels in response to alpha-MSH stimulation. Cell surface expression of F261S MC4R was compared with that of wild-type MC4R. Clinical examinations were performed in subjects carrying F261S mutation and in non-mutated controls.

RESULTSThe alpha-MSH-stimulated reporter gene activity was significantly reduced in cells expressing F261S MC4R, with a maximal response equal to 57% of wild-type MC4R. The F261S mutation also led to a significant change in the Es50 value compared with the wild-type receptor (P<0.01). Immunofluorescent assay revealed a marked reduction in plasma membrane localization of the MC4R in cells expressing the F261S mutant receptor. The resting metabolic rate and fat composition of the mutant carriers were not significantly different from those of the non-mutated obese controls.

CONCLUSIONSThe decreased response to alpha-MSH due to the intracellular retention of MC4R may cause early-onset obesity in the F261S pedigree of Chinese.

Adult ; Age of Onset ; Aged ; Animals ; COS Cells ; Cercopithecus aethiops ; Child ; China ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Obesity ; epidemiology ; metabolism ; Pedigree ; Receptor, Melanocortin, Type 4 ; genetics ; metabolism

OBJECTIVETo prepare oligo microarrays for hepatitis virus detection and genotyping.

METHODSBy analyzing the DNA or cDNA of HBV, HDV and 4 different genotypes of HCV with the BLAST program, a group of specific sequences for the candidate probes was specified. Array Designer 3.0 software was applied to analyze the candidates to select probes with high specificity, identical length and similar melting temperature (Tm). Altogether 16, 8 and 68 oligonucleotide probes were designed for diagnosis of HBV, HDV, and genotyping HCV. Following the synthesizing and purification, oligo probes were deposited on oligonucleotide chips as microarrays for hepatitis virus detection and genotyping. The samples were labeled by RD-PCR method. Hybridization results were analyzed to cross out those probes with low specificity and sensitivity, and those with signal to noise ratios (SNR) less than 4.0.

RESULTSTwo types of gene chips were successfully developed: microarrays for HBV and HDV simultaneous detection and for HCV genotyping.

CONCLUSIONUsing oligo probes to construct gene chips for clinical diagnosis of hepatitis virus is a simple and effective method. It may be widely used in detecting hepatitis viruses and their genotyping in clinical settings.

Base Sequence ; DNA Fingerprinting ; DNA, Viral ; genetics ; Genotype ; Hepatitis B virus ; classification ; genetics ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Oligonucleotide Probes ; Sensitivity and Specificity

BACKGROUND: The recombinant adenovirus has certain toxic reactions to cell growth and survival, but the determination of the optimal multiplicity of infection (MOI) is not well documented. OBJECTIVE: To study the effectiveness and toxicity of transfection of bone marrow mesenchymal stem cells (BMSCs) by adenovirus vectors with different MOIs, and to explore the effect on osteogenic capability of the cells. METHODS: BMSCs at passage 5 were infected with enhanced green fluorescent protein (EGFP) via adenovirus vectors with different MOI (0, 50, 100, 150, 200 and 250). Cell morphology and mortality rate were observed and calculated using trypan blue staining method at 24 hours after cell transfection. The transfection efficiency and the relative mRNA expression of EGFP and Runx2 were analyzed respectively by inverted florescent microscopy and real-time quantitative PCR at 48 hours after cell transfection. The activity of BMSCs transfected with different MOIs was evaluated by MTT, and the optimal MOI was determined thereafter. RESULTS AND CONCLUSION: With the increase of MOI, BMSCs showed decreased adherent ability, and even some cells aged and died. The mortality rate of BMSCs transfected for 24 hours at a MOI of 0 to 250 was 5.80%, 6.67%, 7.95%, 7.76%, 10.35% and 11.18%, respectively, indicating that the mortality rate of BMSCs is positively correlated with the MOI. The transfection efficiency changed insignificantly when the MOI was greater than 100. On the contrary, the cellular viability and osteogenic differentiation capability of rabbit BMSCs were receded when the MOI level was up to 200-250. The study discovered that the suitable scope of MOI to transfect BMSCs is 50-150, and the optimal MOI is 100.

Objective To investigate the clinical effect and safety of butylphthalide injection combined with edaravone injection in the treatment of acute cerebral infarction.Methods A total of 154 patients with acute cerebral infarction were randomly divided into control group (n =77) and treatment group (n =77).Control group was given edaravone injection 30 mg,bid,intravenous drip.Treatment group was given butylphthalide and sodium chloride injection 25 mg,bid,intravenous drip on the basis of control group.All patients were treated for 14 d.The clinical effect,levels of hemorheological indexes,serum endothelin (ET-1),calmodulin (CaM),thromboxane A2 (TXA2),and levels of serum vascular endothelial growth factor (VEGF),VEGF receptor (VEGFR),platelet derived endothelial cell growth factor (PD-ECGF) levels and adverse drug ractions were compared between the two groups.Results After treatment,the total effective rates in treatment group and control group were 84.42％ (65 cases/77 cases) and 70.13 ％ (54 cases/77 cases),with significant difference (P ＜ 0.05).After treatment,the plasma viscosity in treatment group and control group were (1.02 ± 0.14) and (1.29 ± 0.17) mPa · s,whole blood viscosity shear rates were (3.03 ± 0.41) and (3.98 ± 0.56) mPa · s,hematocrit were 0.33 ± 0.04 and 0.42 ± 0.05,platelet aggregation rates were(42.38 ±5.35)％ and(50.43 ± 6.26)％,serum ET-1 were (50.62 ± 6.63) and (62.47 ±8.05) ng · L-1,CaM were(98.65 ± 12.18) and(137.85 ± 16.45) mg · mL-1,TXA2 were (70.02 ± 9.55) and (82.14 ± 11.15)ng · L-1,serum VEGF were (419.36 ± 40.37) and (372.27 ± 39.44) ng · L-1,VEGFR were (405.47 ± 45.36) and (346.56 ± 41.19) ng· L-1,PD-ECGF were (12.47 ± 1.36) and (6.11 ± 0.65) μg · mL-1,all with significant difference (all P ＜ 0.05).The adverse drug reactions were nausea and vomiting,dizziness,drowsiness and heart rate slowing in treatment group and control group.The incidence of adverse drug reactions of treatment group and control group were 14.29％ (11 cases/77 cases) and 10.39％ (8 cases/77 cases),without significant difference (P ＞ 0.05).Conclusion The clinical effect of butylphthalide injection combined with edaravone in the treatment of acute cerebral infarction is exactly,and can effectively improve the indexes of hemorheology,reduce serum levels of ET-1,CaM and TXA2,reduce vascular endothelial damage and prevent thrombosis,increase the levels of serum VEGF,VEGFR,PD-ECGF,and promote neovascularization,with high safety.

Objective To explore a better sputum suction nursing model on the large area burn patients with inhalation injury patients after tracheotomy.Methods Patients with bums area ≥30％ aged 18-60 years,moderate or severe inhalation injury and tracheotomy were included.The patients in the traditional sputum suction group were chosen from January 2009 to October 2010,and received the sputum suction nursing according to the medical nursing technical operation,specific process was as follows:the disposable sputum suction tube was inserted into the airway in certain depth,was slowly lifted from the bottom,and was rotated around in order to completely suck up sputum; and the sputum suction time was on-demand timely.The patients in the improved sputum suction group were chosen from November 2010 to September 2012,and received the planned sputum suction,and the sputum in the tracheal cannula were sucked up for the first times,the length of suction tube insertion was calculated according to the specification of tracheal cannula; the sputum suction tube was changed at the second time,and was inserted into 1-2 cm above tracheal carina; the sputum suction time was after inhalation,call-back or vibration row phlegm,6-8 times/d.Results The pulmonary infection rate in the improved sputum suction group was 30.43％,and 82.35％ in the traditional sputum suction group,and the difference was statistically significant (x2 =15.604,P ＜ 0.01).Conclusions The improved sputum suction nursing method which was planned,inhalation,call-back,etc,is better than the traditional sputum suction,and it is helpful to reduce the pulmonary infection rate in large area burn patients with inhalation injury after tracheotomy.