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

Objective To develop an infusion joint of the two-cavity catheter used for deep vein thrombolysis and explore its clinical application.Methods The joint was composed of a perfusion tube,a perfusion tube tee,two connecting hoses,two connecting hard tubes,two Luer tapers,a liquid inlet connected with the tee,No.1 and No.2 liquid outlets.The liquid medicine was driven to flow through the tube and inlet by gravity,and then came into No.1 and No.2 liquid outlets respectively.Results There were no significant differences between the joint and the traditional way when used for deep vein thrombolysis with a two-cavity catheter (P＞0.05),while the joint had patient satisfaction increased significantly and the time consumed decreased obviously when compared with the traditional way (P＜0.05).Conclusion The joint executes deep vein thrombolysis with a two-cavity catheter and only a set of infusion set,gains advantages in simple structure,low cost and easy operation,and thus is worthy promoting clinically.

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.

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 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

The study is to observe the effect of racemic TJ0711 on blood pressure and heart rate as well as protection of cardiovascular system of renal hypertensive rats after long-term administration. The renal hypertensive models were established by the two-kidney, one-clip (2K1C) method in Wistar rats. Four weeks later, assigned the rats whose SBP had increased at least 4 kPa randomly into 5 groups: racemic TJ0711 10, 20 and 40 mg x kg(-1) groups, carvedilol control group, model group and sham group (n=10), ig administration once daily. The changes of BP (blood press) and HR (heart rate) before and after administration were measured by tail-cuff method weekly. Plasma samples of all animals were taken in 6-8 weeks, and plasma MDA as well as renin, angiotensin II (Ang II) and endothelin-1 (ET-1) levels were measured. Left ventricle was cut off after 9 weeks, and left ventricular weight index (LVWI) and hydroxyproline were measured. The significant decrease of the BP of TJ0711 40 mg x kg(-1) group was observed after TJ0711 ig administration for 4 weeks, and this effect remained till the end of the study. In 8th week, the systolic blood pressure values were: TJ0711 40 mg x kg(-1) group 18.93 +/- 1.82 kPa (vs 21.30 +/- 2.30 kPa, P < 0.05); 20 mg x kg(-1) group 20.68 +/- 3.29 kPa (vs 22.19 +/- 2.88 kPa). The plasma MDA level of all treated groups was significantly lower than that of model group, so were the plasma renin, Ang II and ET-1 levels (P < 0.05). LVWI and hydroxyproline content of myocardial tissue decreased to some extent, but was not significant as compared with that of model group. The study showed that TJ0711 repeated dosing could reduce BP level beginning from drug administration; besides block adrenal alpha and beta receptors to play an antihypertensive role. The sustained antihypertensive effect also related to reduce plasma vasoconstrictor substances and oxidation product MDA. These effects benefited cardiovascular protection.
Angiotensin II ; blood ; Animals ; Antihypertensive Agents ; administration & dosage ; pharmacology ; Blood Pressure ; drug effects ; Endothelin-1 ; blood ; Female ; Heart Rate ; drug effects ; Heart Ventricles ; metabolism ; pathology ; Hydroxyproline ; metabolism ; Hypertension, Renal ; blood ; physiopathology ; Longitudinal Studies ; Male ; Malondialdehyde ; blood ; Organ Size ; drug effects ; Phenoxypropanolamines ; administration & dosage ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar ; Renin ; blood