BACKGROUND:Both high glucose and lipopolysaccharide have been proved to promote the apoptosis of human periodontal ligament fibroblasts (HPLFs), but their interactions on the HPLF apoptosis in vitro have not yet been reported. OBJECTIVE:To investigate the effect of different concentrations of lipopolysaccharide and high glucose on the proliferation, apoptosis and the expression levels of Bax and Bcl-2 in HPLFs in vitro. METHODS:The primarily cultured HPLFs were identified. The 5-8 generations of HPLFs were col ected and used in the subsequent experiment. The HPLFs were cultured in different concentrations of glucose (5.5 and 25 mmol/L) and lipopolysaccharide (0, 1 and 10 mg/L) for 24 and 48 hours, respectively. RESULTS AND CONCLUSION:Lipopolysaccharide (10 mg/L) could significantly inhibit the cel proliferation, promote the cel apoptosis, upregulate the expression levels of Bax and Bcl-2 mRNA and induce a significant decrease in Bcl-2/Bax ratio in the cel s cultured with 5.5 mmol/L glucose (P<0.05). The lipopolysaccharide-induced suppression of cel proliferation, cel apoptosis, the expressions of Bax and Bcl-2 mRNA as wel as decrease in Bcl-2/Bax ratio were significantly strengthened in the HPLFs treated with 25 mmol/L glucose (P<0.05). Analysis of variance found that high glucose and lipopolysaccharide had a significant interaction on the cel apoptosis (P<0.05). These results reveal that lipopolysaccharide-induced suppression of cel proliferation, cel apoptosis and the expressions of Bax and Bcl-2 mRNA are augmented in HPLFs cultured under high glucose condition, indicating lipopolysaccharide and high glucose interactively act in inducing cel apoptosis.

This study aimed at evaluating the quality of the precise powder decoction pieces (PPDP) of L.japonicae Flos (LJF) compared with the traditional commercial slices with chemical fingerprint methods and DNA molecular identification technology.Different specifications of PPDP were prepared,their dry extract contents were in contrast with that of commercial slices.The three batches of commercial slices were collected,and the content uniformity,fingerprint and similarity evaluation before and after the mixing and pulverization were studied by HPLC-DAD and DNA sequence alignment.As a result,the paste rate of PPDP was slightly higher than that of the traditional commercial slices.The dissolution of chlorogenic acid of PPDP was higher than that of the traditional commercial slices.RSD of inter-assay dissolutions of chlorogenic acid of commercial slices was 11.93％,which was reduced to 8.29％ after mixing and preparing into PPDP.The fingerprint showed that the slimilarity of the fringerprint of the mixed and powdered LJF was elevated with 7 common peaks.All the common peaks were increased at different levels.In conclusion,compared with traditional commercial slices of LJF,PPDP apparently improved the dissolution rate and the quality uniformity,indicating that the boiled powder of CRP obviously presented vantages in clinic.

OBJECTIVETo examine the effect of simazine on selected immune parameters in BALB/c mice.

METHODSSimazine (90, 200, 400 mg/kg) was administrered by oral gavage for 21 days in adult BALB/c mice. The negative control group unith distilled water and positive control group administered with cyclophosphamide in abdominal cavity were also established. After the last simazine dose, the mice were sacrificed, and blood, spleens, and thymuses were collected and processed for detection. The relative weight of spleen and thymus was calculated. The rate of T cell in spleen and the concentration of IL-2, IL-4, IgG and IgM were detected by ELISA.

RESULTSThe weights of mice were decreased in 200 mg/kg and 400 mg/kg simazine groups. Thymus and spleen weights were decreased in 200 mg/kg and 400 mg/kg simazine groups compared with the negative control group. The concentration of IL-2, IL-4, IgG and IgM in serum of 200 mg/kg group were (108.50 +/- 3.20) pg/ml, (36.54 +/- 3.36) pg/ml, (46.25 +/- 7.41) μg/ml, (17.58 +/- 2.23) μg/ml respectively;The concentration of IL-2, IL-4, IgG and IgM in serum of 400 mg/kg group were (85.70 +/- 4.00) pg/ml, (35.92 +/- 2.29) pg/ml, (40.08 +/- 6.80) μg/ml, (11.92 +/- 3.23) μg/ml respectively (P < 0.05 or P < 0.01). These results were decreased significantly compared with negative group.

CONCLUSIONSimazine can inhibit the cellular immune function and the humoral immune function.

Animals ; Cytokines ; blood ; Female ; Herbicides ; toxicity ; Immunity, Cellular ; drug effects ; Immunity, Humoral ; drug effects ; Immunoglobulin M ; blood ; Male ; Mice ; Mice, Inbred BALB C ; Simazine ; toxicity ; T-Lymphocytes ; drug effects

Adolescent ; Brain Neoplasms ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Humans ; Melanoma ; metabolism ; pathology ; Melanoma-Specific Antigens ; metabolism ; Neurilemmoma ; metabolism ; pathology ; S100 Proteins ; metabolism ; Vimentin ; metabolism

Aim To pick out the siRNA which could most effectively inhibit the expression of TLR4 in microglial cells and to detect the cytotoxicity of the transfection complex.Methods Five siRNAs were chemicaly synthesized:four of them were used to inhibit TLR4 expression in microglial cells,the rest was fluorescence-labeled mismatch siRNA as a nagative control.They were all transfected into microglial cells,respectively.TLR4 mRNA was detected 24 h after transfection by RT-PCR and its protein expression wasobserved by Western blot 48 h later.The cytotoxicity of complex was detected using MTT.Results ① The transfection rate was high enough in microglial cells with siRNA(40 pmol)and LipofectamineTM 2000(1 μl).② The TLR4 siRNA pool reduced TLR4 mRNA by 85%(siRNA_(439)),73%(siRNA_(312)),67%(siRNA_(1495))and 33%(siRNA_(2062))respectively compared with mismatch siRNA-treated group 24 h after transfection in a microglial cell line.③ The TLR4 siRNA439 was the most effective siRNA(P<0.01).④ The cell survival rates were above 85% in the groups of Lipofectamine~(TM) 2000 1 μl compound less than 40 nmol·L~(-1) siRNA.Conclusions ① The TLR4 siRNA_(439) can inhibit TLR4 expression most effectively in microglial.② 40 nmol·L~(-1) siRNA and 1 μl Lipofectamine~(TM) 2000 have low cytotoxicity,which are suitable for transfection.

Objective To construct a recombinant immunotoxin expression vector composed of a single-chain Fv fragment of Sehistosorna japomicum and PE38KDEL gene,and identify the binding activity of the purified product with SEA antigen.Methods The V_H and V_L genes were amplified by PCR from the parent monoclonal antibody NP11-4.Then the amplified scFv and PE38KDEL genes were inserted into the expression vector pBAD/gIII A.The fusion protein expressed in E.coli Top10F' induced by L-arabinose.After purification,the activity of the immunotoxin was evaluated by Westem-blot and ELISA.Results The new recombinant immunotoxin expression vector pBAD/gIII A-scfv-PE38KDEL was constructed successfully.The main product was in inclusion bodies.ELISA assay showed that the refolding recombinant immunotoxin remained binding activity with SEA antigen.Conclusion A new recombinant expression plasmid pBAD/gIII A-scfv-PE38KDEL has been constructed and expressed successfully,which is useful in further study of the treatment of schistosomiasis japonica.

OBJECTIVETo analyze cell-free fetal DNA in maternal plasma for prenatal screening of beta-thalassaemia major.

METHODSSix couples undergoing prenatal diagnosis of beta-thalassaemia (gestational age range 23-26 weeks) were enrolled in this study. The husbands were all carriers of the CD17 (A-->T) mutation, and the wives carried another beta-thalassaemia mutation. The allele-specific primers and two fluorescent cycling probes were synthesized for the detection of the CD17 (A-->T) mutation, using FAM and HEX fluorescence labeling, respectively. The cell-free fetal DNA in the maternal plasma was detected using real-time PCR, and the fetal genotype was confirmed by cord blood conventional prenatal diagnosis.

RESULTSIn the 6 pregnancies, FAM and HEX fluorescent signals were detected in 3 maternal plasma samples; in the other 3 samples, only FAM fluorescent signals were detected, suggesting the absence of paternally derived CD17 (A-->T) mutation.

CONCLUSIONExamination of cell-free fetal DNA in maternal plasma using real-time PCR and cycling probe technology can be effective means for prenatal screening of beta-thalassaemia major.

Adult ; DNA ; blood ; DNA Mutational Analysis ; DNA Probes ; Female ; Fetal Diseases ; blood ; diagnosis ; genetics ; Heterozygote ; Humans ; Maternal-Fetal Exchange ; Point Mutation ; Pregnancy ; Prenatal Diagnosis ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; beta-Thalassemia ; blood ; diagnosis ; genetics

This paper introduces the current development and challenges of vision prosthesis.
Prosthesis Design ; Visual Prosthesis

Objective To evaluate the changes in expression of NF-κB, IL-6 and TNF-α in spinal cord in a rat model of bone cancer. Methods Seventy-two female SD rats weighing 150-180 g were randomly divided into 3 groups (n = 24 each): control group (group C);sham operation group (group S) and bone cancer pain group (group BP). Bone cancer was induced by intra-tibial inoculation of 1 × 105 Walker 256 breast cancer cells. Paw withdrawal threshold to mechanical stimulation was measured with yon Frey filaments. The expression of NF-κB p65, IL-6 and TNF-α mRNA in the spinal cord was determined by RT-PCR and the expression of NF-κB p65 by immuno-histochemistry and NF-κB p65 positive cell count was determined. Results The paw withdrawal threshold was significantly lower and the expression of NF-κB p65, NF-κB p65 mRNA, IL-6 mRNA, TNF-α mRNA and NF-κB p65 positive cell count in the spinal cord were significantly higher in group BP than in group C and S ( P ＜0.05 or 0.01 ). Conclusion Intra-tibial inoculation of Walker 256 breast cancer cells activates NF-κB in the spinal cord, leading to the increased release of IL-6 and TNF-α and mechanical hyperalgesia.

Adenocarcinoma ; genetics ; Adult ; Aged ; Female ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Mutation ; Receptor, Epidermal Growth Factor ; genetics