Human sTNFR1 (soluble tumor necrosis factor receptor 1) gene was amplified by RT-PCR from Hela cells. A recombinant expression vector of sTNFR1-MBP was constructed in pMAL-c2x, and transformed into E. Coli JM109.It was sequenced and confirmed to be identifical to the sTNFR1 gene in data bank. Recombinant protein sTNFR1-MBP was induced by IPTG and purified by Amylose resin Affinity Chromatography. sTNFR1-MBP was binded to sTNFR1's antibody in Western-blotting. From MTT assays, the results showed that sTNFR1-MBP could effectively block the cytotoxicity mediated by TNF?on QSG7701 cells. Annexin V-FITC staining and flowcytometry were used to observe the recombinant protein's anti-apoptosis capacity and the recombinant protein has marked anti-apoptosis effect in vitro.sTNFR1-MBP had good biological activity and it will be employed in further study.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Ascites ; pathology ; Biopsy ; Biopsy, Fine-Needle ; Child ; Cytodiagnosis ; methods ; Diagnosis, Differential ; Female ; Humans ; Leukemia-Lymphoma, Adult T-Cell ; pathology ; Lymphoma, B-Cell ; pathology ; Lymphoma, Large B-Cell, Diffuse ; pathology ; Lymphoma, T-Cell ; pathology ; Male ; Middle Aged ; Pleural Effusion ; pathology ; Young Adult

Dioscin has a wide range of biological effects and broad application prospects. However the studies concerning the toxicology and mechanism of dioscin is small. This article is to study the hepatotoxicity of dioscin and the effect of dioscin treatment on expression of aryl hydrocarbon receptor (AhR) mRNA and CYP1A mRNA and protein in HepG2 cells in vitro. Dioscin 0.5-32 µmol · L(-1) exposed to HepG2 cells for 12 h, cell viability was examined by CCK-8 assay and the release rate of lactate dehydrogenase (LDH) was to evaluate cell membrane damage. HepG2 cells morphologic changes were quantified by inverted Microscope, and the effect on production of reactive oxygen species (ROS) was detected by flow cytometry. The mRNA expression of CYP1A and AhR was evaluated by RT-RCR. The protein expression of CYP1A1 was detected by western blot. The cell viability was significantly inhibited after HepG2 cells were exposed to dioscin 0.5-32 µmol · L(-1). Compared with the control, the LDH release rate and ROS were significantly increased. The expression of CYPlA and AhR mRNA was increased. The expression of CYP1Al protein was increased after dioscin treatment, and resveratrol, an AhR antagonist, could downregulate the expression of CYP1A1. It follows that large doses dioscin has potential hepatotoxicity. The possible mechanism may be dioscin can active aryl hydrocarbon receptor (AhR) and induce the expression of CYP1A.
Cell Survival ; drug effects ; Chemical and Drug Induced Liver Injury ; etiology ; Cytochrome P-450 CYP1A1 ; genetics ; Diosgenin ; analogs & derivatives ; toxicity ; Hep G2 Cells ; Humans ; L-Lactate Dehydrogenase ; secretion ; RNA, Messenger ; analysis ; Reactive Oxygen Species ; metabolism ; Receptors, Aryl Hydrocarbon ; genetics

OBJECTIVETo introduce a new method using calcium phosphate cement/Danshen drug delivery system for treating ischemic necrosis of the femoral head and evaluate its curative effect.

METHODSFrom May 2000 to June 2005, 48 adult patients (54 hips) with ischemic necrosis of the femoral head at Stages I, II and III of antigen reactive cell opsonization (ARCO) were treated with implantation of calcium phosphate cement/Danshen drug delivery system in the involved femoral head. The operation consisted of removal of the necrotic bone under weight-loading cartilage and the implantation of phosphate cement/Danshen drug delivery system, and all manipulations were made percutaneously through a bone tunnel in the trochanter. The functions of the hip joint were evaluated and X-ray films were taken preoperatively and postoperatively.

RESULTSPostoperative follow-up was 45.5 months on average, ranging from 27 to 78 months. According to the evaluation criterion of "Dandong 1995" for therapeutic effect of adult ischemic necrosis of the femoral head, the therapeutic effects were excellent in 33 hips, good in 17, fair in 3 and poor in 1, with the excellent and good rate of 92.6%.

CONCLUSIONSThis method is relatively simple with little invasion. It not only improves the microcirculation of the femoral head by local application of traditional Chinese medicine, but also provides mechanic buttress in the weight-loading area to prevent collapse during repairing, which is beneficial to repair and reconstruction of femoral head. It may be a choice of minimal invasion surgery for ischemic necrosis of the femoral head at Stages I, II and III of ARCO.

Adult ; Aged ; Bone Cements ; Calcium Phosphates ; administration & dosage ; Decompression, Surgical ; Drug Delivery Systems ; Female ; Femur Head ; blood supply ; Femur Head Necrosis ; diagnostic imaging ; therapy ; Humans ; Ischemia ; therapy ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Radiography ; Salvia miltiorrhiza

Animals ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Hepatocytes ; pathology ; Liver Cirrhosis, Experimental ; pathology ; Rats ; Rats, Wistar ; Serum

OBJECTIVESTo investigate the internal links between immune responses and Tregs and cytokine by the expression of T regulatory cells (Tregs), Foxp3 mRNA of different response groups and the detection of cytokine secretion after hepatitis B vaccination.

METHODSBlood samples were collected in different response groups. Real-time fluorescence quantitative PCR was used to detect the expression of Foxp3 mRNA of peripheral blood mononuclear cells; The surface markers CD4 and CD25 in peripheral-blood mononuclear cells were determined by flow cytometry; ELISA tests were used to detect the production level of phytohemagglutinin (PHA) in peripheral blood mononuclear cells, IL -4, IL-12, IL-18 stimulated by HBsAg and (IFN) gamma.

RESULTS(1) Foxp3 expressions in response group and non-response group were higher before or after PHA and HBsAg were stimulated. Differences were statistically significant (P value less than 0.05) ; (2) In peripheral blood, the percentage of CD4+CD25+ Treg of CD4+ T cells in response group (0.59%+/-0.46%) was obviously lower than those in control group (1.30%+/-1.44%) ; (3) Peripheral blood mononuclear cells stimulated by PHA and HbsAg in each group, the concentration of IFNgamma in non-response group [(11.00+/-9.03) IU/ml] was markedly lower than those in response group [(38.88+/-28.16) IU/ml],and differences were statistically significant (P value less than 0.01); (4) In PHA- or HBsAg-stimulated peripheral-blood mononuclear cells, the concentrations of IL-18, IL-4 and IL-12 had no significant difference.

CONCLUSIONSTo some extent, CD4+CD25+Foxp3+ Treg cells may be involved in negative regulation of the immune responses to HBV vaccination. Immune non-response to HBV vaccination may be connected to insufficient secretion of IFNgamma; There was no correlation between the titer of anti-HBs and the expressions of IFNgamma and CD4+CD25+ Foxp3.

Adolescent ; Antibody Formation ; CD4 Antigens ; metabolism ; Female ; Forkhead Transcription Factors ; immunology ; Hepatitis B ; immunology ; prevention & control ; Hepatitis B Vaccines ; immunology ; Hepatitis B virus ; immunology ; Humans ; Interferon-gamma ; immunology ; Interleukin-12 ; immunology ; Interleukin-18 ; immunology ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Interleukin-4 ; immunology ; Male ; T-Lymphocytes, Regulatory ; immunology ; Young Adult

To research the effect of Ginseng Radix et Rhizoma and Aconiti Lateralis Radix Praeparata compatibility on cardiac toxicity in rats by UPLC-Q-TOF/MS, and explore the endogenous markers and molecule mechanism. Different compatibility of Shenfu decoction were given to male Wistar rats at dosage of 20 g · kg(-1) for 7 days, collected the serum, and analyze the endogenous metabolites effected by Shenfu formulation by principal component analysis and partial least-squares analysis. Results showed that content of glutathione, phosphatidylcholine and citric acid decreased in mixed-decoction group, while ascorbic acid, uric acid, D-galactose, tryptophan, L-phenylalanine increased. The results showed cardiac toxicity of Aconiti Lateralis Radix Praeparata in Shenfu mixed-decoction. Shenfu co-decoction group showed a similar or weaker trend compared with control group, but most of them do not have a statistically significant. The results indicated the scientific basis of Shenfu compatibility by comparison of co-decoction group with mixed-decoction group. Shenfu compatibility can reduce cardiac toxicity induced by Aconiti Lateralis Radix Praeparata, and citric acid, glutathione, phosphatidyl choline, uric acid might be regarded as potential markers of cardiotoxicity.
Animals ; Biomarkers ; Cardiotoxicity ; Drugs, Chinese Herbal ; toxicity ; Glutathione ; blood ; Least-Squares Analysis ; Male ; Metabolomics ; methods ; Principal Component Analysis ; Rats ; Rats, Wistar

To study the effect of Siwu decoction on the function and expression of P-glycoprotein (P-gp) in Caco-2 cells. The Real-time quantitative poly-merase chain reaction (Q-PCR) was used to analyze the mRNA expression of MDR1 gene in Caco-2 cells. Flow cytometer was used to study the effect of Siwu decoction on the uptake of Rhodamine 123 in Caco-2 cells, in order to evaluate the efflux function of P-gp. Western blotting method was used to detect the effect of Siwu decoction on the P-gp protein expression of Caco-2 cells. Compared with the blank control group, after Caco-2 incubation with Siwu decoction at concentrations of 3.3, 5.0, 10.0 g x L(-1) for 24, 48, 72 h, the mRNA expression of MDR1 was up-regulated, suggesting the effect of Siwu decoction in inducing the expression of MDR1. After the administration with Siwu decoction in Caco-2 cells for 48 h, the uptake of Rhodamine 123 in Caco-2 cells decreased by respectively 16.6%, 22.1% (P < 0.05) and 45.4% (P < 0.01), indicating that the long-term administration of Siwu decoction can enhance the P-gp efflux function of Caco-2 cells. After the incubation of Caco-2 cells with Siwu decoction for 48 h, the P-gp protein expression on Caco-2 cell emebranes, demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp.
ATP Binding Cassette Transporter, Sub-Family B ; genetics ; metabolism ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Caco-2 Cells ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Up-Regulation ; drug effects

Molecular mechanisms whereby H2S influences its targets have been of intriguing interest. In this work, L-lactic dehydrogenase ( L-LDH) was used as the protein target, and three kinds of H2S-donor reagents ( NaHS, Na2S, and polysulfide) were chosen. The interactions of these H2S-donor reagents with L-LDH were disclosed by molecular fluorescent assays for real-time monitoring of L-LDH activity. The results of the SDS-PAGE showed that H2S might not interact with L-LDH to form disulfide/trisulfide bonding. Circular dichroism spectra assays revealed that H2S reagents could be likely to react with cysteine thiols to yield sulfurated thiol (-SSH) derivatives in L-LDH, and sulfur-containing PS ( polysulfide) was a stronger protein S-sulfurating agent than the other two sulfides. Matrix assisted laser desorptionionization time-of-flight tandem mass spectrometry ( MALDI-TOF-MS/MS) study showed partial S-sulfuration of the active cysteine sites existed in L-LDH. In conclusion, H2S exerts its biological effects as a gasotransmitter through its reactions with cysteine thiols in proteins by S-sulfuration.

Objective To explore the transmission chain of COVID-19 by serum antibody detection, and to provide scientific evidence for the prevention and control of the epidemic. Methods Field epidemiological investigation was used to determine the COVID-19 cases and their close contacts. The 2019-nCoV nucleic acid in throat swabs and anal swabs were examined by RT-PCR. Serum specimens were collected for anti-2019-nCoV IgM antibody detection and combined IgM/IgG detection. Results Case A had no confirmed exposure to COVID-19. However, case C and D had dinner and lived together with case A; they reported contact history and dinner history with other confirmed COVID-19 cases(H, L). Case A tested positive for 2019-nCoV nucleic acid, whereas case C and D were negative. Moreover, case A and C were IgM antibody positive, while case D was negative. Case A, C and D were all positive for combined IgM/IgG. In addition, case D had clinical symptom, while case C did not. Conclusion Serum antibody detection can be used as an effective supplement to the inference of transmission chain of COVID-19, which may facilitate determining the source of infection and improving the evidence.