Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; metabolism ; T-Lymphocytes, Regulatory ; metabolism ; Th17 Cells ; metabolism

This study was purposed to investigate the efficacy of different whole flow lysis reagents for lysis of red blood cells in flow cytometric analysis. The expression of immunocytes was detected by flow cytometry after lysis of red blood cells using commercial reagents (Optilyse C, FACS Lysing Solution) and self-made red blood cell lysis reagents (RBC Lysis Buffer), the detection results were analyzed comparatively. The results showed that there was no significant difference in the percentage of CD3e(+), CD3e(+)CD4(+), CD3e(+)CD8a(+), CD3e(-)CD19(+), CD3e(-)NK1.1(+) and Gr-1(+) cells between 3 different lysis reagent groups. However OptiLyse C solution was suitable to Gr-1(+) cell detection, but did not suit to Foxp3(+) Treg detection. The self-made RBC Lysis Buffer and FACS Lysing Solution were suited to Foxp3(+) Treg detection. It is concluded that the use of self-made RBC Lysis Buffer for flow cytometry can get the lysis efficiency of commercially available lysis solutions when samples are prepared in accordance with standardized procedure. The self-made RBC Lysis Buffer not only can satisfy experimental requirements, but also can reduce the experimental costs.
Animals ; Erythrocyte Count ; Erythrocytes ; immunology ; metabolism ; Flow Cytometry ; instrumentation ; methods ; Immune System ; immunology ; Indicators and Reagents ; analysis ; Mice ; Mice, Inbred C57BL

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.

Objective:To study the effects of compound schisandra extracts (CSE) (schisandra,astragalus,acanthopanax,and rhodiola)on the exhaustive swimming time and the levels of blood urea nitrogen(BUN),serum lactic acid(LD),liver glycogen and muscle glycogene,superoxide dismutase(SOD)activity,and malonaldehyde(MDA) level in the mice and to charify its anti-fatigue effect and the mechanism.Methods:Eighty male ICR mice were randomly divided into blank control group,50 mg·kg-1 CSE group,100 mg·kg-1 CSE group,and 200 mg·kg-1CSE group;there were 20 mice in each group.The mice were administered orally for 30 d.Then 10 mice were randomly selected for exhaustive swimming test in each group and the exhaustive swimming time of the mice was recorded.The remaining 10 mice in each group were used for 90 min swimming,then all the mice were sacrificed and the blood and tissue samples were taken for the measurement of the levels of BUN,LD,liver glycogen and muscle glycogen,the SOD activity and MDA level;the total inhibitory rate of oxidation of CSE in vitro was determined by linoleic acid-ferric thiocyanate method.Results:Compared with blank control group,the exhaustive swimming time of the mice in 50,100,and 200 mg·kg-1 CSE groups were significantly increased (P<0.01);the levels of BUN and LD of the mice in 100 and 200 mg·kg-1 CSE groups were significantly decreased (P<0.05 or P<0.01);the levels of liver glycogen and muscle glycogen of the mice in 100 and 200 mg·kg-1 groups were significantly increased(P<0.05 or P<0.01);whereas the SOD activities were significantly increased and the levels of MDA were decreased(P<0.05 or P<0.01).Compared with 100 mg·kg-1 CSE group,the levels of serum BUN and LD of the mice in 200 mg·kg-1 CSE group were decreased (P<0.01),and the levels of liver glycogen and muscle glycogen were increased(P<0.05).The total inhibitory rate of oxidation of 5 g·L-1CSE was 76.94%.Conclusion:CSE has an anti-fatigue effect and the mechanism may be related to anti-oxidation effect.

OBJECTIVETo examine the effect of exposure to extremely low-frequency electromagnetic fields (ELF EMFs) on the liver function of workers.

METHODSThe workers in a factory were selected as subjects, and the recent physical examination data of these workers were collected. The workers aged 20∼40 years and with more than 2 years' working experience were included for analysis; considering the intensity of electromagnetic field, the workers exposed to less electromagnetic radiation were assigned to exposure I group (n = 123), those exposed to more electromagnetic radiation to exposure II group (n = 229), and those not exposed to electromagnetic radiation to control group (n = 212). There were no significant differences in sex, age, height, and body weight between the three groups (P > 0.05). Physical examination, including measurements of direct bilirubin (DBil), alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (GGT), and albumin, was performed in a health examination center. The intensity of electromagnetic field was measured by EFA-300 power frequency electromagnetic field analyzer, and the intensity of noise by AWA5610D integrating sound level meter.

RESULTSThe intensities of electric field and the magnetic field in exposure II group were significantly higher than those in the exposure I group. The levels of ALT, ALP, AST, GGT and albumin in exposure II group were significantly higher than those in exposure I group and control group. However, the level of direct bilirubin in exposure II group was significantly lower than that in exposure I group and control group.

CONCLUSIONOccupational exposure to ELF EMFs may affect human liver function.

Adult ; Alanine Transaminase ; blood ; Aspartate Aminotransferases ; blood ; Bilirubin ; blood ; Electromagnetic Fields ; adverse effects ; Female ; Humans ; Liver ; physiopathology ; Male ; Occupational Exposure ; adverse effects ; Young Adult

BACKGROUNDIt is known that excessive release of glutamate can induce excitotoxicity in neurons and lead to seizure. Dexamethasone has anti-seizure function. The aim of this study was to investigate glutamate-dexamethasone interaction in the pathogenesis of epilepsy, identify differentially expressed genes in the hippocampus of glutamate-induced epileptic rats by mRNA differential display, and observe the effects of dexamethasone on these genes expression.

METHODSSeizure models were established by injecting 5 microl (250 microg/microl) monosodium glutamate (MSG) into the lateral cerebral ventricle in rats. Dexamethasone (5 mg/kg) was injected intraperitoneally at 30 minutes after MSG inducing convulsion. The rats' behavior and electroencephalogram (EEG) were then recorded for 1 hour. The effects of dexamethasone on gene expression were observed in MSG-induced epileptic rats at 1 hour and 6 hours after the onset of seizure by mRNA differential display. The differentially expressed genes were confirmed by Dot blot.

RESULTSEEG and behaviors showed that MSG did induce seizure, and dexamethasone could clearly alleviate the symptom. mRNA differential display showed that MSG increased the expression of some genes in epileptic rats and dexamethasone could downregulate their expression. From more than 10 differentially expressed cDNA fragments, we identified a 226 bp cDNA fragment that was expressed higher in the hippocampus of epileptic rats than that in the control group. Its expression was reduced after the administration of dexamethasone. Sequence analysis and protein alignment showed that the predicted amino acid sequence of this cDNA fragment kept 43% identity to agmatinase, a member of the ureohydrolase superfamily.

CONCLUSIONSThe results of the current study suggest that the product of the 226 bp cDNA has a function similar to agmatinase. Dexamethasone might relax alleviate seizure by inhibiting expression of the gene.

Animals ; Base Sequence ; Dexamethasone ; pharmacology ; Electroencephalography ; drug effects ; Epilepsy ; chemically induced ; drug therapy ; genetics ; Gene Expression Profiling ; Gene Expression Regulation ; drug effects ; Male ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Rats ; Rats, Sprague-Dawley ; Sodium Glutamate ; pharmacology

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

OBJECTIVETo evaluate the possible association of the MTHFR C677T polymorphism with genetic susceptibility to hepatocellular carcinoma (HCC) in a Chinese population.

METHODSFive hundred and eight HCC cases and 543 controls were studied. The MTHFR genotypes were determined using a PCR-based restriction fragment length polymorphism (RFLP) method. Odds ratios (ORs) for HCC and 95% confidence intervals (CIs) from unconditional logistic regression models were used to evaluate relative risks. Potential HCC risk factors were included in the logistic regression models as covariates in the multivariate analyses on genotypes and HCC risks.

RESULTSNo overall significant difference in genotype distribution was found when comparing all HCC cases to controls (P = 0.258). However, a significantly increased risk of HCC was observed among T/T homozygotes (adjusted OR = 1.66, 95% CI = 1.08-2.54, P<0.05) compared to C-allele carriers (CC or CT). When stratified with sex, this trend was more prominent in females, but not in males. Females who were homozygous (T/T) for the C677T polymorphism were at a 2.64-fold (95% CI = 1.19-5.88, P<0.05) increased risk of developing HCC when compared to C-allele carriers. However in males, T/T homozygotes had a similar risk with C-allele carriers.

CONCLUSIONThe MTHFR C677T polymorphism may be associated with a higher HCC risk in females, but not in males in this population.

Aged ; Carcinoma, Hepatocellular ; genetics ; Female ; Genetic Predisposition to Disease ; Humans ; Liver Neoplasms ; genetics ; Logistic Models ; Male ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Middle Aged ; Polymorphism, Genetic

OBJECTIVETo evaluate the efficacy and safety of live combined Bifidobacterium, Lactobacillus and Enterococcus capsules in treatment of irritable bowel syndrome.

METHODSEighty-five patients [male 32, female 53; age (45.31+/-11.72) years] were given live combined Bifidobacterium, Lactobacillus and Enterococcus capsules 1260 mg/d t.i.d.x4 weeks. Syndrome scales were used to evaluate the efficacy in gastrointestinal syndrome. Fecal flora was also measured before and after the treatment. Six bacteria were cultured and the colony forming units were counted in stool. SPSS was used for data analysis.

RESULTSSeventy-four patients finished the follow-up. No side-effect was found. For treatment of irritable bowel syndrome, the effective rate of live combined Bifidobacterium, Lactobacillus and Enterococcus capsules was 56.8% in the second week, 74.3% in the fourth week and 73.0% in the sixth week. Single symptom was improved, especially in abdominal pain and stool character. The probiotica containing live combined Bifidobacterium, Lactobacillus and Enterococcus could increase bifidobacterium count (P<0.01) and lactobacillus count (P<0.05); decrease bacteroides count (P<0.05) and enterococci count (P<0.01); No obvious changes were observed in clostridium difficile colonitis and enterobacteriaceae (P>0.05).

CONCLUSIONThe result of the study indicated that the administration of live combined Bifidobacterium, Lactobacillus and Enterococcus improved the symptom of irritable bowel syndrome and that there was a gradual increase of this effect. Thereafter conditions remained stable for 2 weeks. That improvement may be associated with alterations in gastrointestinal flora.

Adult ; Bifidobacterium ; Enterococcus ; Female ; Humans ; Intestines ; microbiology ; Irritable Bowel Syndrome ; drug therapy ; Lactobacillus ; Male ; Middle Aged ; Probiotics ; therapeutic use

BACKGROUNDBecause of lacking skills in scanning the normal fetal facial structures and their corresponding ultrasonic features, misdiagnoses frequently occur. Therefore, we studied the appearance features and improved displaying skills of fetal facial anatomy in order to provide basis for prenatal diagnosis.

METHODSTwenty fetuses with normal facial anatomy from induced labor because of other malformations except facial anomalies were immersed in a water bath and then scanned ultrasonographically on coronal, sagittal and transverse planes to define the ultrasonic image features of normal anatomy. The coronal and sagittal planes obtained from the submandibular triangle were used for displaying the soft and hard palate in particular.

RESULTSFacial anatomic structures of the fetus can be clearly displayed through the three routine orthogonal planes. However, the soft and hard palate can be displayed on the planes obtained from the submandibular triangle only.

CONCLUSIONSThe superficial soft tissues and deep bony structures of the fetal face can be recognized and evaluated by routine ultrasonographic images, which is a reliable prenatal diagnostic technique to evaluate the fetal facial anatomy. The soft and hard palate can be well demonstrated by the submandibular triangle approach.

Face ; diagnostic imaging ; embryology ; Humans ; In Vitro Techniques ; Ultrasonography