Antineoplastic Agents ; administration & dosage ; therapeutic use ; Child ; Female ; Humans ; Injections, Intralesional ; Interferon-alpha ; administration & dosage ; therapeutic use ; Male ; Neoplasms ; drug therapy ; Recombinant Proteins ; Treatment Outcome

The rice gall dwarf disease, caused by the Rice gall dwarf virus (RGDV) is a serious disease occurring in rice in many regions of Guangdong province. As a basis to control the disease we have studied the genomic diversity of a variety of isolates from different locations. Genome segment 8(S8), encoding a main outer capsid protein (Pns8) of RGDV five isolates (BL, CH, DQ, GZ, XY) from Guangdong province was cloned and sequenced. The results revealed that all the S8 segments of the five isolates consisted of 1 578 nucleotides and had a single open reading frame (ORF) extending for 1 301 nucleotides from nucleotide 21 which encoded a polypeptide of 426 amino acids with an estimated molecular weight of 47.4 kDa. The S8 full-length sequence and the ORF sequence shared 97.3%-98.8% and 97.3%-99.1% nucleotide sequence identities within the five Chinese isolates, and shared 94.8%-95.6% and 95.0%-96.0% identities with those of the Thailand isolate respectively. The deduced amino acid sequence of Pns8 in GZ isolate was identical to that in the Thailand isolate, while the amino acid sequence variability of Pns8 within five Chinese isolates ranged from 0.5% to 2.1%. These results indicate that the S8 segment of RGDV is highly conserved in different isolates from different locations. The S8 cDNA from the XY isolate was cloned into the plasmid vector pET-28b(+) and a fused expression protein with an apparent molecular mass of 51kDa was specifically detected in an analysis of Escherichia coli Rossetta(DE3)Ⅱcells. To our knowledge, this is the first report on analysis of the RGDV segment 8 sequence and genetic comparison of different RGDV isolates and their protein expression.

OBJECTIVETo evaluate the genotypic diversity of Streptococcus sobrinus (Ss) between children suffering with severe early childhood caries (S-ECC) and caries-free children by arbitrarily primed-polymerase chain reaction (AP-PCR).

METHODSA total of 178 children aged from 42 to 54 months were recruited from 14 urban kindergartens. The S-ECC group contained 87 children with more than 5 decayed teeth, and the control group was composed of 91 caries-free children. Stimulated whole saliva was collected by chewing paraffin. All mutans streptococcus isolates were subcultured, biochemically characterised and identified by PCR as Streptococcus mutans (Sm) and Ss. Then the Ss isolates were genotyped by AP-PCR.

RESULTSThe frequency of Ss detection was 18% in S-ECC children, which was significantly higher than 3% in caries-free children (P < 0.01). Twenty-two distinct genotypes of Ss were identified from 53 clinical isolates. In S-ECC group, one to three genotypes of Ss were detected in each saliva sample. Only one genotype of Ss was detected in all the caries-free children. One genotype of Ss were shared by three S-ECC children. The genotypes of isolates in S-ECC group were relate to decayed-missing-filled teeth (r = 0.50, P < 0. 05).

CONCLUSIONSThe rate of Ss detection was significantly higher in S-ECC children than in caries-free children. Isolates of Ss displayed genetic polymorphism. The multi-genotypes of Ss was related to differences in caries susceptibility. Strains of Ss with same genotype were present in unrelated subjects.

Child, Preschool ; Dental Caries ; microbiology ; Female ; Genotype ; Humans ; Male ; Polymorphism, Genetic ; Streptococcus sobrinus ; genetics ; isolation & purification

Objective To observe the effects of 75 gram glucose oral tolerance test (75 g OGTT) and standard mixed meal test (SMMT) on insulin secretion function of the islets of Langerhans and plasma free fatty acid (FFA) in patients with type 2 diabetes mellitus.Methods Seventy-six patients with type 2 diabetes without using insulin and with no obvious complications were recruited for 75 g OGTT following overnight fasting on the first day and SMMT (bread 50 g,egg 50 g and milk 250 ml) on the 7th day.Blood specimens were collected from each patients before the tests and 30 min,60 min,120 min and 180 min after glucose or meal load to measure their levels of plasma glucose,serum insulin,C peptide,FFA and lipids (total cholesterol,triglyceride,high-density and low-density lipoprotein cholesterol).Results No difference in fasting plasma glucose,serum insulin,C peptide,FFA and lipids between 75 g OGTT and SMMT was found.Postprandial plasma glucose 30 min,60 min,120 min and 180 min after 75 g OGTT was significantly higher than that after SMMT,with (15.3?3.5) vs (9.9?3.4) mmol/L,(18.2?4.8) vs (12.8?4.0) mmol/L,(16.3?5.8) vs (12.2?4.9) mmol/L and (10.6?5.4) vs (9.5?4.5) mmol/L (F=28.1,P

To investigate the effects of interferon alpha-2b on proliferation and apoptosis in HL-60 cells, HL-60 cells were cultured in different concentrations of IFN alpha-2b. The morphologic changes were observed by Wright's and acridine orange (AO) and ethidium bromide (EB) staining respectively. Inhibition of proliferation was detected by MTT. Expression of CD13(+) was checked by indirect fluoroimmunoassay. The results showed that apoptosis rate of HL-60 cells assayed by the above-mentioned two methods was (51 +/- 2)% and (78 +/- 3)% respectively and OD(570) values of proliferation inhibited were 1.8 +/- 0.1 and 1.0 +/- 0.1 respectively when the concentrations of the IFN(alpha-2b) were 500 and 10,000 U/ml in culture for 48 hours. Morphology and count of CD13(+) cells were changed. CD13(+) cell expression rate was (62 +/- 2)% and (30 +/- 3)% respectively when the concentrations of the IFN(alpha-2b) were 500 and 10,000 U/ml in culture for 48 hours. It is concluded that IFN(alpha-2b) can enhance the apoptosis of HL-60 cells, inhibit their proliferation, promote their maturation and differentiation.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; CD13 Antigens ; biosynthesis ; genetics ; Cell Proliferation ; drug effects ; HL-60 Cells ; Humans ; Interferon-alpha ; pharmacology ; Recombinant Proteins

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.

BACKGROUNDCarbon disulfide (CS(2)) is a commonly used organic solvent. Many epidemiological investigations and animal experiments have indicated that learning and memory ability can be affected to different degrees after long-term exposure to CS(2), but the mechanisms are still unclear. The aim of this study was to explore the possible mechanisms of CS(2)-related impairment of the learning and memory ability of rats, by investigating the effects of CS(2) on nitric oxide synthase (NOS) activity and NOS mRNA expression in rat hippocampus.

METHODSRat models of toxicity were generated by inhalation of various doses of CS(2). After two months of inhaling intoxication, the activities of constitutive NOS (cNOS) and induced NOS (iNOS) in the hippocampus were measured. The levels of neuronal NOS (nNOS) mRNA and iNOS mRNA were measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR).

RESULTScNOS activity was significantly decreased compared with controls, while iNOS activity was changed only slightly. CS(2) treatment significantly decreased nNOS mRNA levels. iNOS mRNA levels were significantly increased only at higher doses of CS(2).

CONCLUSIONThe effect of CS2 on learning and memory ability in rats is related to the activity of NOS and the expression of nNOS in the hippocampus.

Animals ; Carbon Disulfide ; pharmacology ; Gene Expression Regulation, Enzymologic ; drug effects ; Hippocampus ; drug effects ; metabolism ; Nitric Oxide Synthase ; genetics ; metabolism ; RNA, Messenger ; Random Allocation ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Spectrophotometry

OBJECTIVETo investigate the early clinical detection and new method for the treatment of congenital dislocation of hip in infants.

METHODSFrom 2006 to 2010, 95 infants with congenital dislocation of hip were treated with self-made pygal cloth sling, including 25 males and 70 females, with an average age of 3.2 months old ranging from 0 to 6 months. Some patients were detected incidentally for the symptoms like asymmetric muscle strength or lower limbs range of motion, and all the patients got diagnosed with dislocation.

RESULTSAfter the treatment, all of the patients received outpatient view once a month and taken X-ray examination bimonthly. Pygal cloth sling was removed after 2 months. According to the assessment criteria made by LIU Yuan-zhong, 90 patients got an excellent result, 2 good, 2 fair and 1 poor.

CONCLUSIONTreatment of congenital dislocation of hip in infants with self-made pygal cloth sling promotes the development of acetabulum and femoral head, and worthy further clinical applications.

External Fixators ; Female ; Hip Dislocation ; therapy ; Humans ; Infant ; Infant, Newborn ; Male

OBJECTIVETo investigate the clinical features of thyroid disease occurring in response to antiviral therapy in patients with chronic hepatitis C (CHC).

METHODSEighty-two patients diagnosed with CHC were recruited for study from our hospital between 2009 and 2010. All patients were given a 48-week course of antiviral combination therapy with pegylated-interferon (Peg-IFN; 180 mug qw ih) and ribavirin (RBV; 15 mg/kg bw). Patient sera was collected prior to treatment (baseline), at treatment weeks 24 and 48, and post-treatment week 24, and used to detect changes in levels of thyroid function markers, thyroid-specific and other autoantibodies, complement factors, and immunoglobulins (Igs). Differential expression of biomarkers was assessed between patients who developed thyroid disorder and those who did not.

RESULTSAt treatment week 48, 13.4% (11/82) of cases developed hypothyroidism, 3.7% (3/82) developed hyperthyroidism, 20.7% (17/82) tested positive for thyroglobulin antibody, and 22.0% (18/82) tested positive for thyroid peroxidase antibody. The patients who did not develop thyroid disease had significantly higher post-treatment levels (vs. baseline) of IgG (14.84 +/- 2.61 vs. 12.95 +/- 3.32 g/L, F = 10.458, P = 0.002) and C4 (0.26 +/- 0.09 vs. 0.22 +/- 0.08 g/L, F = 6.835, P = 0.011) and significantly lower IgM (0.86 +/- 0.48 vs. 1.00 +/- 0.42 g/L, F = 9.106, P = 0.003). The patients who developed thyroid disease showed no significant differences in the baseline and post-treatment levels of IgG, C4, or IgM. When the two groups of patients who did or did not develop thyroid disease were compared, there was no difference in the amount of patients who achieved sustained virological response.

CONCLUSIONAntiviral-induced thyroid disease in patients with refractory hepatitis C manifests as clinically-detectable abnormalities in serum levels of thyroid autoantibody and markers of hypothyroidism. Levels of other autoantibodies and Igs do not correlate with the development of thyroid disease in these patients, and thyroid disease does not appear to affect the efficacy of Peg-IFN + RBV antiviral therapy.

Antiviral Agents ; therapeutic use ; Drug Therapy, Combination ; Hepatitis C, Chronic ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Polyethylene Glycols ; therapeutic use ; Ribavirin ; therapeutic use ; Thyroid Diseases ; chemically induced

Objective　To investigate morphological changes of endothelial cells after nordihydroguaiaretic acid (NDGA) treatment in vitro. Methods　The morphological changes of human umbilical vein endothelial cells (HUVEC) cell line ECV-304 and the cell apoptosis rate in sub-G0 phase were observed with invert, light and electron microscope and flow cytometry after NDGA treatment at different concentrations or with PBS (0.01 mol/L) as control. Results　①After the treatment of NDGA at 50～200 μmol/L for 1～3 d or up to 8 d at 100 μmol/L, ECV-304 cells tended to elongate into a shuttle-like sparse appearance and those in mitosis were decreased, indicating the suppression of cell proliferation. All these alteration was in a time-and dose-dependent manner. ②NDGA-treated ECV-304 cells displayed morphological features of apoptosis, especially at the 48th h after the treatment. With flow cytometry, the cells in sub-G0 phase were significantly increased, and reached its peak at hour 12 (20.42%) after NDGA treatment. In addition, the degeneration and necrosis of ECV-304 cells were related to the concentrations of NDGA. Conclusion　NDGA can inhibit the proliferation and growth of endothhelial cells, and induce apopotosis, which might also inhibit angiogenesis.