To investigate me material basis of Mahuang Fuzi Xixin decoction (MFXD) for anti-inflammation and immune-suppression based on the combined method of serum chemical and serum pharmacological. The LC-MS/MS fingerprints of MFXD, drug-containing serum and blank serum were compared to define the components in plasma. Histamine, β-hexosaminidase released from RBL-2H3 cell infulenced by drug-containing serum at different time points were measured by ELISA. The effect of drug-containing serum on lipopolysaccharide-induced splenocyte proliferation at different time points were determined by MTT. A correlation analysis was made on components of MFXD and pharmacological indexes based the stepwise regression method. After the intragastrical administration with MFXD, 32 components were discovered in rat serum, including 27 prototype components (10 from Mahuang, 13 from Fuzi and four from Xixin) and five unknown components. Compared with blank serum, drug-containing serum could reduce the release of histamine from RBL-2H3 induced by antigen at different time points (P < 0.05); except the 4-hour drug-containing serum, all of the remaining drug-containing serums could inhibit the RBL-2H3 mastocyte degranulation induced by antigen at different time points (P < 0.05). Drug-containing serum could significantly lipopolysaccharide-induced mouse splenocyte proliferation at 15 and 30 min (P < 0.05). A regression analysis was made on the chemical data of components absorbed into blood and pharmacological indexes, i. e. release rate of histamine, release rate of β-hexosaminidase and inhibition rate of splenocyte. This suggested the close correlations among methyl pseudo-ephedrine, pseudoephedrine and histamine released from RBL-2H3 induced by antigen; pseudoephedrine, hypaconine, methyl pseudoephedrine and β-hexosaminidase released from RBL-2H3 induced by antigen; as well as benzoyl hypaconine, benzoylaconine, 14-benzoyl-10-OH-mesaconine, mesaconine and lipopolysaccharide-induced mouse splenocyte proliferation. Methylpseudoephedrine, pseudoephedrine, benzoyl hypaconine, benzoylaconine and mesaconine may be part of material basis of MFXD on anti-inflammation and immune suppression.
Animals ; Anti-Inflammatory Agents ; chemistry ; pharmacology ; Cell Degranulation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Histamine ; immunology ; Immunosuppressive Agents ; chemistry ; pharmacology ; Male ; Mass Spectrometry ; Mast Cells ; drug effects ; immunology ; Mice ; Rats ; Rats, Wistar ; Serum ; chemistry

Objective To investigate the clinical effect of of rt-Pa Early Thrombolytic Therapy on the Patients with Acute Cerebral Infarction (ACI). Methods 40 patients with acute cerebral infarction were randomly divided into rt-PA group and control group, 20 cases in each group. The treatment time window was less than 3 hours. On the basis of the routine medication of cerebral infarction, rt-PA group was given rt-PA intravenous thrombolytic therapy, the control group was treated with sodium ozagrel. The National Institutes of Health Stroke Scale (NIHSS) and Barthel index (BI) were used to evaluate the efficacy and safety. Results The NIHSS scores of two groups after treatment were significantly improved compared with before treatment, rt-PA group improved significantly better than the control group (P<0.05); At 21 days of treatment, the excellence rate of rt-PA group was 80% and the ratio of BI≥95 was 40.00%, the excellence rate of control group was 25.0% and the ratio of BI≥95 was 20.00%, the rt-PA group was significantly higher than the control Group (P<0.01); there was no significant difference between the two groups in the mortality rate, the incidence of secondary cerebral hemorrhage and vascular restenosis in the two groups. Conclusion Within 3 hours of onset of ACI, compared with the sodium ozagrel therapy, the rt-PA intravenous thrombolytic therapy is more safe and effective in the treatment of ACI patients, and it is worthy to be widely used in clinical treatment.

Objective To observe expression of the keratin 10 (K10), c-myc and cox2 in buccal mucous membrane exfoliated cells, and changes of liver function of coal-burning arsenism patients, in order to explorate coal-burning arsenic poison circumference biology symbol. Methods The buccal mucous membrane exfoliated cells were collected from both arsenism patients and normal controls. Real-time PCR was employed to detect the changes of K10, c-myc, cox2 genes expression in these cells. Simultaneously, circumference venous blood was extracted and examinated liver function. Results K10 was found obviously overexpressed in arsenism patients(3.60±0.94) compared to control(1.82±0.68), the difference had statistics significance(t=2.15, P<0.05), c-mye and cox2 did not found obviously changed(c-myc: 3.50±2.77,3.39±2.07; cox2:5.90±1.40,4.73±1.91; t=1.26,1.65, P> 0.05). The serum ALT of patients(25.83±2A5) obviously increased than control(36.86±1735, t=2.55, P<0.05). Conclusions Expression of K10 gene in buccal mucous membrane cells may be regarded as sensitive molecular markers for skin pathologic changes in arsenic patients. The liver is a sensitive target organ of inorganic arsenic.

OBJECTIVETo investigate the effects of Shadu Cao Mixture (SDCM, traditional Chinese medicine) on immune functions of immunosuppression mice.

METHODSFifty BALB/C mice were randomly divided into blank control group, model group, SDCM low-dose, middle-dose and high-dose group. Except the blank control group, other groups were intraperitoneal injected with cyclophosphamide (40 mg/kg) to establish immunosuppression mice model. The blank control group and model group received gavage administration with nonnal saline, while the other groups received gavage administration with different doses of SDCM (10, 20, 40 m/kg for 15 days) respectively. The number of leukocytes and serum levels of interleukin-2 (IL-2), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in peripheral blood, spleen index, and the function of NK cells were measured.

RESULTSCompared with the model group , SDCM increased the number of leukocytes and serum concentrations of IL-2, TNF-α and IFN-γ in peripheral blood and improved the spleen index and the function of NK cells significantly (P < 0.05-0.01).

CONCLUSIONSDCM could remarkably enhance the immune functions of immunosuppression mice induced by cyclophosphamide.

Animals ; Cyclophosphamide ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Immunosuppression ; Interferon-gamma ; blood ; Interleukin-2 ; blood ; Killer Cells, Natural ; immunology ; Mice ; Mice, Inbred BALB C ; Spleen ; immunology ; Tumor Necrosis Factor-alpha ; blood

AIMTo investigate the antagonistic effects of the novel compounds on vasoconstriction induced by ET-1 and the effect on the blood pressure of stroke-prone spontaneously hypertensive rats.

METHODSOrgan bath experiment and whole cardiac function experiment were used.

RESULTSThe analogues of o-CPhe-D-Trp-D-Phe(-X)-OH showed good ability against endothelin biological effects. When X was displaced by 3-F, 3-Cl or 4-Cl, the novel compounds inhibit the vascular constriction induced by ET-1 in a concentration-dependent manner, the IC50 +/- L95 were (0.09 +/- 0.05), (0.15 +/- 0.06) or (0.11 +/- 0.03) mumol.L-1 respectively. The blood pressure of stroke-prone spontaneously hypertensive rats was decreased. No significant effect on cardiac function of rats was discovered.

CONCLUSIONThe results demonstrate that among the six kinds of compounds, those with o-CPhe-D-Trp-D-Phe (-X)-OH configuration showed good biological effects.

Animals ; Aorta ; drug effects ; Blood Pressure ; drug effects ; Endothelin Receptor Antagonists ; Endothelins ; pharmacology ; Hypertension ; drug therapy ; physiopathology ; Male ; Molecular Structure ; Peptides ; chemical synthesis ; chemistry ; pharmacology ; therapeutic use ; Rats ; Rats, Inbred SHR ; Rats, Wistar ; Structure-Activity Relationship ; Vasoconstriction ; drug effects

OBJECTIVETo provide evidence for illustrating the molecular mechanism of nickel carcinogenesis, and to identify the differential expression of protein in crystalline NiS-induced neoplastic transformation of human bronchial epithelial cell by proteomics technology.

METHODSTwo dimensional electrophoresis (2-DE) and the ImageMaster 3.10 software were used to analyze the differential expression of protein, matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) combined with database search was applied to identify protein peroxiredoxin 2 (PDX2) related to malignant transformation.

RESULTSThe good 2-DE pattern including resolution and reproducibility was obtained. Nearly 700 expressed proteins per 2-D gel were isolated with molecular weights (MW) ranging from 14,400 to 94,000 KD and pI 3 - 10. A protein PDX2 with MW 21,890 KD, pI 5.66, which was highly expressed in malignantly transformed cell, was identified using MALDI-TOF-MS.

CONCLUSIONPDX2 was involved in malignant transformation of human bronchial epithelial cell induced by crystalline nickel sulfide.

Bronchi ; cytology ; Cell Line ; Cell Transformation, Neoplastic ; chemically induced ; metabolism ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Nickel ; toxicity ; Peroxiredoxins ; metabolism ; Proteome

OBJECTIVETo explore the role of EGF in regulating HaCaT apoptosis through peroxisome proliferator-activated receptor beta (PPARbeta).

METHODSCultured HaCaT cells were divided into different groups with different additives in culture medium as control (normal culture), TNF-alpha (with addition of 10 ng/mL TNF-alpha), EGF (with addition of 20 ng/mL EGF), EGF + TNF-alpha (cells were treated with 10 ng/mL TNF-alpha for 60 mins after the exposure to 20 ng/mL EGF for 4 hs) groups. Conjugation activity and transcription activity of PPARbeta of HaCaT cells in each group were detected by electrophoretic mobility shift assay (EMSA) and luciferase gene analysis (LGA). Protein expression of PPARbeta of HaCaT cells after transfected by missense oligonucleotide (scrODN) and antisense oligonucleotide (asODN) was determined by Western blot. Caspase-3 activity and apoptosis rate were detected by flow cytometry.

RESULTSConjugation and transcription activity of PPARbeta DNA were enhanced as shown in EMSA and LGA. Compared with that of cells in groups transfected by scrODN, protein expression of PPARbeta in cells of groups transfected by asODN was obviously inhibited as shown in Western blot. Caspase-3 activity of cells in TNF-alpha and EGF + TNF-alpha groups transfected by asODN was stronger than that of cells in TNF-alpha and EGF + TNF-alpha groups transfected by scrODN (P < 0.01). Apoptosis rate of cells in control, EGF, TNF-alpha, and EGF + TNF-alpha groups which were transfected by scrODN was (7.31 +/- 0.45)%, (7.43 +/- 0.21)%, (39.78 +/- 0.65)%, (28.34 +/- 0.54)% respectively, and that in those groups transfected by asODN was (8.22 +/- 0.51)%, (7.83 +/- 0.67)%, (46.78 +/- 0.48)%, (44.69 +/- 0.83)%. Apoptosis rate of cells in TNF-alpha and EGF + TNF-alpha groups transfected by asODN was respectively higher than that in TNF-alpha and EGF + TNF-alpha groups transfected by scrODN (P < 0.01).

CONCLUSIONSEGF inhibits HaCaT KC apoptosis caused by TNF-alpha in a PPARbeta-dependent manner.

Apoptosis ; drug effects ; Cell Culture Techniques ; Cell Line ; Epidermal Growth Factor ; pharmacology ; Humans ; PPAR-beta ; genetics ; metabolism ; Transcription, Genetic ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors

Construction of mutant strain is an essential method in pathogenesis researches. The conventional method for Brucella unmarked deletion mutant construction is based on suicide plasmid, but the efficiency is very low. In the present study, we first optimized the electroporation parameters, and then, the cloning plasmid pEX18Gm containing sacB was successfully used to construct unmarked deletion mutant of the type IV secretion system. This indicated that by using conventional cloning plasmid as suicide plasmid in Brucella, unmarked deletion mutants can be constructed with high efficiency.

OBJECTIVETo investigate the influence of antisense phosphorothioate oligonucleotides on peroxisome proliferator-activated receptors (PPARbeta) in the TNF-alpha mediated apoptosis of HaCat cells.

METHODSHaCat cells were resuscitated and randomly divided into normal control (without transfection), sham (merely with liposome transfection), scrODN (with transfection of 4 micromol/L PPARbeta scrODN), asODN (with transfection of 4 micromol/L PPARbeta asODN), TNF-alpha with transfection of 10 micromol/L TNF-alpha), scrODN + TNF-alpha with 10 micromol/L TNF-alpha stimulation after transfection of 4 micromol/L PPARbeta scrODN), asODN + TNF-alpha with 10 micromol/L TNF-alpha stimulation after transfection of 4 micromol/L PPARbeta asODN) groups. The mRNA and protein levels of PPARbeta were determined with RT-PCR and Western blotting, respectively. The changes in cell morphology were observed with Hoechst 33258 fluorescent staining to quantitate apoptotic rate of nuclei. The effect of PPARbeta asODN on HaCat cell viability was assayed with MTT method. Activation of caspase-3 was evaluated with caspase colorimetric analysis kit.

RESULTSThe mRNA and protein expression of PPARbeta in normal control, sham, scrODN groups were similar, but it decreased obviously in asODN group. The nuclear apoptotic rate in normal control, scrODN and asODN groups were rather low, and the caspase-3 activity in these groups was also low. After 24 hours of culture, the nuclear apoptotic rate in TNF-alpha and scrODN + TNF-alpha groups were (33.1 +/- 2.7)% and (32.9 +/- 3.0)%, respectively, while that in asODN + TNF-alpha group was obviously increased (58.8 +/- 4.6)%, with the caspase-3 activity significantly higher, but the number of live cells markedly lower than that in the former 2 groups (P < 0.05).

CONCLUSIONPPARbeta expression can promote the apoptosis of HaCat cells mediated by TNF-alpha.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Cycle ; Cell Line ; Cell Proliferation ; Humans ; Oligonucleotides, Antisense ; genetics ; pharmacology ; PPAR-beta ; genetics ; pharmacology ; RNA, Messenger ; metabolism ; Transfection ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo study the effect and mechanism of saffor injection on renal ischemia/reperfusion (I/R) injury in rats.

METHODSeventy-five SD rats were randomly divided into five groups (n = 15, in each), normal control groups, I/R control groups, low-dose treatment groups, middle-dose treatment groups and high-dose treatment groups. After rat's I/R injury model was established, renal function was assessed by measuring serum creatinine, blood urea nitrogen, urine osmotic pressure and urine osmotic pressure/blood osmotic pressure, the apoptosis rate in I/R renal tissure was measured by TUNEL method and caspase-3 concentration was measured by immunohistochemistry.

RESULTReperfusion of the ischemic kidney induced marked renal dysfunction. Saffor injection significantly inhibited the reperfusion-associated increase in apoptosis rate and caspase-3 protein absorbance value. Moreover, the renal dysfunction at all treatment groups was markedly ameliorated by Saffor injection. (P < 0.01).

CONCLUSIONThe results show that saffor injection significantly reduces the renal dysfunction and injury caused by I/R of the kidney, And the protective effect of Saffor injection may be related to the inhibition of cell apoptosis and caspase-3 gene expression following renal I/R.

Animals ; Apoptosis ; drug effects ; Blood Urea Nitrogen ; Carthamus tinctorius ; chemistry ; Caspase 3 ; metabolism ; Creatinine ; blood ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Female ; Injections ; Kidney ; blood supply ; Male ; Osmotic Pressure ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; blood ; enzymology ; pathology