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.

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

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

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

The study was purposed to investigate whether processing and storage conditions might influence the stability of the HCV RNA in whole blood or in plasma. The samples obtained from seven patients known to be positive for HCV RNA were kept in different storage conditions with different anticoagulants, and at the end of processing the plasma samples were frozen at -80 degrees C until fluorescent quantitative PCR testing. The results showed that there was no significant loss of HCV RNA titers in whole blood anticoagulated with CPDA or ACD or EDTA or none (P > 0.05), while differences in comparison of the EDTA-anticoagulant storage condition with three other anticoagulants storage conditions at 4 degrees C after 48 hours were significant (P < 0.05). The HCV RNA level decreased to 53.8%, 72.5% and 29.8% after 48 hours of storage of whole blood anticoagulated with ACD at 4 degrees C, 25 degrees C and 37 degrees C respectively. The HCV RNA level of plasma samples stored at 4 degrees C and at 25 degrees C (room temperature) after 7 days decreased to 70.9% and 25.1% respectively. After four freeze-thaw cycles the HCV RNA level decreased 38.9% in plasma samples. It is concluded that the HCV RNA is stable relatively. The HCV RNA is resistant to degradation under routine laboratory handling and storage conditions or blood collection, transport and processing conditions. The influence of different anticoagulants on the stability of HCV RNA is different. Blood samples would better be stored at 4 degrees C after collection and plasma separated within 48 hours. And it is important for the stability of HCV RNA undergoing asepsis blood collection process. HCV RNA remains stable at 4 degrees C for at least 7 days or at room temperature for 3 days, allowing greater flexibility in samples collection and transport in transfusion practice nowadays. HCV RNA in plasma samples subject to up to three short-term freeze-thaw cycles is still stable.
Blood Donors ; Blood Preservation ; methods ; Hepacivirus ; genetics ; Hepatitis C ; virology ; Humans ; RNA, Viral ; blood ; drug effects ; Specimen Handling ; standards ; Temperature ; Time Factors

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

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 seek a sequential method for the management of residual wounds in burn patients.

METHODSThree chronic residual wounds on each of 25 burn patients were either covered with vaseline gauze (A group), human tissue-engineered active skin (Active Skin, B group) or Active Skin after rinsing with fluid containing oxygen and vacuum assisted drainage ( C group) on wounds. The contents of (TNF)a in granulation tissue were assayed by enzyme linked immunosorbent assay (ELISA). Expression of metalloproteinase-13 (MMP-13) mRNA in granulation tissue was determined with reverse transcription polymerase chain reaction (RT-PCR). Moreover, quantity of wound bacteria in the wounds and wound healing rate were determined with usual method.

RESULTSThe quantities of wound bacteria in C group on 3,6,9, 12 post-treatment day( PTD) were (5.30 +/- 1.60), (1.30 +/-0.80) , (1.70 +/- 0. 60)and (0.60 +/-0. 10)clone formation unit/ml( CFU/ml) , respectively, which were obviously lower than those in A and B groups. The contents of TNFa and expression of metalloproteinase-13 (MMP-13) mRNA in granulation tissue in C group on 6 PTD were [ (0. 650 +/- 0. 040) ng/mg and 0. 210 +/- 0. 010,] ,respectively, and they were evidently lower than those in A group [(1.550 +/-0. 370)ng/mg,1. 040 +/- 0. 050, P <0.01] and B group (0. 810 +/- 0.080) ng/mg, 0.640 +/- 0.030, P <0.01]. Meanwhile, the contents of (TNF)a and expression of MMP-13 mRNA in B group were also obviously lower than those in A group. The wound healing ratio in C group on 15 and 30 PTD were markedly higher than those in A or B group ( P <0.01).

CONCLUSIONCovering the residual burn wounds with Active Skin after rinsing with fluid containing oxygen followed by vacuum assisted drainage can improve repairing of residual burn wounds.

Adolescent ; Adult ; Burns ; microbiology ; therapy ; Female ; Humans ; Male ; Matrix Metalloproteinase 13 ; metabolism ; Middle Aged ; Negative-Pressure Wound Therapy ; RNA, Messenger ; metabolism ; Skin, Artificial ; Therapeutic Irrigation ; Tissue Engineering ; Tumor Necrosis Factor-alpha ; metabolism ; Wound Healing ; Young Adult

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.