Objective To explore the possible mechanism and protective effect of Xuebijing injection (血必净注射液)and dexamethasone on rats with paraquat-induced chronic pulmonary injury.Methods Thirty male Wistar rats were randomly divided into six groups:normal group(n=5),model group(n=5), treatment groups(n=20).In the normal group,normal saline was used,while in the other groups,20% paraquat 80 mg/kg was injected peritoneally for poisoning.After 2 hours of intoxication,low dose Xuebijing injection(1.25 g/kg),high dose Xuebijing injection(2.50 g/kg),dexamethasone(25 mg/kg),high dose Xuebijing injection combined with dexamethasone(combined group)respectively were administered into the four different treatment groups,equal amount of normal saline was given to the normal and model groups,and the treatment continued for 4 days.At 28 days after paraquat injection,5 rats in each group were killed respectively,serum transforming growth factor-?1(TGF-?1)and hydroxyproline(HYP)level in the lung homogenate were measured,and pulmonary coefficient and histological changes were observed.Results In the treatment groups,the levels of serum TGF-?1 and lung tissue HYP,pulmonary coefficient were leas than those of model group,and among the treatment groups,combined group had the best results(all P

Litsea cubeba is one of aromatic medicinal plant belonging to family Lauraceae. The roots, stems and fruits of L. cubeba have been widely applied as folk medicines in some districts in China for relieving rheumatism and cold, regulating Qi (meridian) to alleviate pain. Previous studies revealed that this species contains major alkaloids, in specific aporphines, and minor flavonoids, lignans as well. Related pharmacological investigations demonstrated its activities and clinical applications on cardiovascular diseases, anti-cancer, against rheumatoid arthritis, relieving asthma and anti-allergic effects, as anti-oxidants, and so on. As an effort for further exploration of this bioactive ingredients and potential drug development, this paper summarizes most phytochemical and pharmacological results. Further, future prospects are also included.
Animals ; Drug Therapy ; Humans ; Litsea ; chemistry ; Molecular Structure ; Plant Extracts ; chemistry ; pharmacology ; Plants, Medicinal ; chemistry

A phytochemical investigation on the aerial parts of a Tibetan medicine Meconopsis horridula, by solvent extraction, repeated chromatographies on silica gel, Sephadex LH-20, and preparative TLC techniques, led to the isolation of 9 compounds. By spectroscopic analysis and comparison of its 1H and 13C-NMR data with those in literatures, their structures were identified as oleracein E(1), N-( trans-p-coumaroyl) tyramine (2), chrysoeriol (3), apigenin (4), hydnocarpin (5), p-coumaric acid glucosyl ester (6), stigmast-5-ene-3beta-ylformate (7), 3beta-hydroxy-7alpha-ethoxy-24beta-ethylcholest-5-ene (8), and beta-sitosterol (9), respectively, among which compounds 6-8 were isolated from the genus for the first time,and 1,3 were isolated from the species for the first time. A MTT method was applied to evaluate the cytotoxic activity of compounds 14 against the human hepatocellular liver carcinoma cell line (HepG2), and compound 1 showed significant cytotoxicity against HepG2,with its inhibitory rate of 52.2% at 10 micromol x L(-1).
Medicine, Tibetan Traditional ; Molecular Structure ; Papaveraceae ; chemistry ; Plant Extracts ; chemistry ; Spectrometry, Mass, Electrospray Ionization

A phytochemical investigation on the roots and stems of Litsea cubeba led to the isolation of seven isoquinolone alkaloids. By spectroscopic analysis and comparison of their 1H and 13C-NMR data with those in literatures, these alkaloids were identified as (+)-norboldine (1), (+)-boldine (2), (+)-reticuline (3), (+)-laurotetanine (4), (+)-isoboldine (5), (+)-N-methyl-laurotetanine (6), and berberine (7), respectively. Among them, 7 was isolated from the genus for the first time. The evaluation of these compounds showed weak anti-inflammatory activity against NO production in RAW 267.4 and BV-2 cells.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Litsea ; chemistry ; Molecular Structure ; Plant Roots ; chemistry ; Plant Stems ; chemistry ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo investigate the effect of mito chondrial K(ATP) channels (mitoK(ATP)) inhibitor 5-hydroxydecanoate(5-HD) on chronic hypoxic pulmonary artery hypertension (CHPAH) rats and its underlying mechanisms.

METHODSForty-eight male SD rats were equally divided into 4 groups randomly (n=12): normal group, hypoxia group, hypoxia + 5-HD group, hypoxia + Diazoxide group. Except the first group, the other three groups were put into hypoxic [O2 (10.0% +/- 0.3%] and nonrmobaric chamber for four weeks to establish chronic hypoxic model and received different interference. When the interference completed, right heart catheter was used to detect the mean pulmonary arterial pressure (mPAP) of each rat and PKC-alpha mRNA expression in pulmonary arteries was detected by reverse transcription-polymerase chain reaction (RT-PCR) and protein expression by Western blot.

RESULTS(mPAP was much higher in hypoxia group than that in normal group (P < 0.01) while in hypoxia + 5-HD group and hypoxia + diazoxide were decreased significantly compared to hypoxia group (P < 0.01). (2) The protein and mRNA levels of PKC-alpha in the hypoxic group were higher than those in normal group (P < 0.05).

CONCLUSION5-HD plays a protective role on CHPAH. The mechanism of its effect may be attributed to inhibiting MitoK(ATP).

Animals ; Decanoic Acids ; pharmacology ; Hydroxy Acids ; pharmacology ; Hypertension, Pulmonary ; etiology ; metabolism ; physiopathology ; Hypoxia ; complications ; physiopathology ; Male ; Muscle, Smooth, Vascular ; metabolism ; Potassium Channel Blockers ; pharmacology ; Potassium Channels ; drug effects ; Protein Kinase C-alpha ; genetics ; metabolism ; Pulmonary Artery ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective:To observe the effect of electroacupuncture (EA) on the expressions of acetylcholine (ACh) and mucin 5AC (MUC5AC) in the lungs of rats with chronic obstructive pulmonary disease (COPD),and explore the mechanism of EA in treating COPD.Methods:Thirty Sprague-Dawley (SD) rats were randomly divided into a control group,a COPD group,and an EA group,with 10 rats in each group.The control group was a group of normal rats.The COPD rat model was induced by cigarette smoke combined with lipopolysaccharide (LPS).The COPD rats were treated with EA at bilateral Feishu (BL 13) and Zusanli (ST 36) in the EA group,30 min each time,once a day,successively for 14 d.The lung function was tested.The contents of ACh and MUC5AC in lungs and bronchoalveolar lavage fluid (BALF) were detected by enzyme-linked immunosorbent assay (ELISA).Pearson method was used to analyze the correlation between pulmonary function and the content of MUC5AC in lungs.The mRNA and protein expressions of MUC5AC in lung tissues were detected by real-time polymerase chain reaction (RT-PCR) and Western blot (WB),respectively.The immune response of MUC5AC was observed by immunohistochemistry.Results:Eight rats were left in each group,and the other two died.Compared with the control group,the total airway resistance (Raw) increased significantly and dynamic compliance (Cdyn) decreased significantly in the COPD group (P＜0.01);compared with the COPD group,the Raw level declined significantly and Cdyn increased significantly in the EA group (P＜0.01).The contents of ACh and MUC5AC in the lungs and BALF were remarkably higher in the COPD group compared with those in the control group (P＜0.01,P＜0.001);compared with the COPD group,the contents of ACh and MUC5AC were significantly lower in the EA group (P＜0.05,P＜0.001).There was a negative correlation between MUC5AC content and lung function (P＜0.001).The mRNA and protein expressions of MUC5AC in the lungs were significantly higher in the COPD group than in the control group (P＜0.001);compared with the COPD group,the expressions were significantly lower in the EA group (P＜0.01).Compared with the control group,the immune response of MUC5AC in the airway epithelium significantly increased in the COPD group (P＜0.001);the immune response of MUC5AC was significantly lower in the EA group compared with that in the COPD group (P＜0.001).Conclusion:EA treatment can improve the lung function of COPD rats,which may be related to its effect in the down-regulation of ACh and MUC5AC contents in the lungs as well as the inhibition of mucus hypersecretion.

BACKGROUNDTo construct the full-length complementary DNA of HCV genome from an HCV infected patient.

METHODSFour HCV gene fragments (1.6, 3.5, 2.4 kb and 2.6 kb) were amplified by RT-PCR from serum of a Chinese patient and fused and connected together to produce a 9.2 kb subgenomic fragment, which was further cloned into a cassette vector with fixed 5-prime and 3-prime termini of HCV to make the full-length cDNA. The cDNA heterogeneity was analyzed by comparing the sequences of 4 isolated HVR1 regions. The prokaryotic expressed Core, NS3 protease, NS3 helicase were detected for their specific reactivities with patient serum by Western blot analysis. And the protease activity of NS3 was evaluated in a cell-based NS3/4A-SEAP expression system.

RESULTSThe cDNA covered the near full-length of HCV genome from the patient's serum. The difference among HVR1 regions indicated no selection of HCV variants during RT-PCR and the quasi-species characteristic of the amplified cDNA. The prokaryotic expressed viral proteins could be identified by patient serum. In the NS3/4A-SEAP system, NS3 could cleave the 4A-4B site between NS4A and SEAP proteins and resulted in the secretion of SEAP in culture media.

CONCLUSIONThese results suggest that the cloned HCV cDNA encodes a complete and functional open reading frame and will be useful for further construction of infectious cDNA clone.

Base Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Genome, Viral ; Hepacivirus ; genetics ; Hepatitis C ; virology ; Humans ; Molecular Sequence Data ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo elucidate the variation in characterizations and genetic evolution of the matrix protein 2 or ion channel protein(M2) genes of avian influenza subtype H5N1 viruses in the boundary region of Yunnan province from 2008 to 2012.

METHODSA total of swab samples were collected from foreign poultry such as the junction between Yunnan and Vietnam, Laos,myanmar and wild birds in boundary region of Yunnan province from 2008 to 2012 and screened by H5N1 subtype-specific multiplex RT-PCR. The M genes of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis of M2 genes were performed with sequences of the known reference strains.

RESULTSA total of 71 positive samples were found out of 1240 samples and the positive rate was 5.72%. A total of 14 different M2 sequences were obtained from 30 positive samples and were divided into 3 distinct clades or sub-clades(1.2.1, 1.2.2 and 2) by phylogenetic analysis, 5, 7 and 2, respectively. The M2 genes and Hemagglutinin(HA) genes of H5N1 viruses from the boundary region of Yunnan province had showed different relationship of genetic evolution. The substitution or mutation of key amino acids sites had been found among the domains of epitope, adamantane-resistance, and poultry or human original viral strains.

CONCLUSIONThe M2 genes of H5N1 subtype viruses in boundary region of Yunnan province from 2008 to 2012 showed genetic divergence and the virus of clade 1.2.2 had become dominant epidemic strain in this region.

Animals ; Birds ; virology ; Chickens ; virology ; China ; Evolution, Molecular ; Influenza A Virus, H5N1 Subtype ; classification ; genetics ; Influenza in Birds ; virology ; Phylogeny ; Poultry ; virology ; Viral Matrix Proteins ; genetics

OBJECTIVETo elucidate the characteristics of variation and the genetic evolution of non-structural protein (NS1, NS2) genes related to avian influenza subtype H5N1 viruses isolated from the boundary region of Yunnan province.

METHODSSwab samples were collected from foreign poultry and wild birds in the boundary regions of Yunnan province and screened by H5/N1 subtype-specific multiplex RT-PCR. The NS segment of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis on those available NS1, NS2 genes were performed with sequences of the known reference strains.

RESULTS71 positive samples were identified from 1240 samples, with the positive rate as 5.72%. Fourteen different NS segment sequences were obtained from 30 representative positive samples and could be divided into 3 distinct clades or sub-clades (I-1, I-2 and II), by phylogenetic analysis. The NS1/NS2 genes and Hemagglutinin (HA) genes of H5N1 viruses from the boundary regions of Yunnan province showed different relationships regarding the characteristics on genetic evolution. The substitution or mutation of key amino acids sites had been noticed in the nuclear location signal domains, effect domain, and other pathogenicity markers.

CONCLUSIONNS genes of H5N1 subtype viruses in boundary region of Yunnan province showed genetic divergence and the virus of clade I-2 and II had become dominant epidemic strains in this region since 2010.

Amino Acid Sequence ; Animals ; Animals, Wild ; Birds ; virology ; China ; epidemiology ; Evolution, Molecular ; Genome, Viral ; Influenza A Virus, H5N1 Subtype ; genetics ; isolation & purification ; Influenza in Birds ; epidemiology ; virology ; Phylogeny

OBJECTIVETo study the Th17/Treg (regulatory T cells) immunoregulation in patients coinfected with TB and HIV before and after HAART(highly active anti-retroviral therapy).

METHODS10 HIV cases coinfected with TB (HIV/TB group) and 10 cases infected with HIV only (HIV group) received HAART. PBMCs were stained and immunophenotyping of Th17 (IL-17 expressing T cells) and CD4+ CD25 T cells (Treg) were analysed by flow cytometry.

RESULTSThe pre-treatment patients tended to have lower Th17 cells and higher Tregs cells compared to post-treatment (1.90% +/- 0.9% vs. 4.65% +/- 1.48%, 16.48% +/- 4.91% vs. 8.29% +/- 3.13% respectively). The percentage of IL-17 before and after HAART were 1.90 +/- 0.9% vs. 4.65 +/- 1.48% respectively in HIV/TB group patients (P < 0.01). The difference between the percentage of IL-17 before and after HAART in the HIV/TB group and the HIV group were 2. 65 +/- 1.62% vs. 0.67% +/- 0.46% respectively (P < 0.01). IL-17 expressing T cells were increased faster after HAART in the former group than the latter. The percentage of Treg before and after HAART were 16.48% +/- 4.91% vs. 8.29% +/- 3.13% respectively in HIV/TB group (P < 0.01). The difference between the percentage of Treg before and after HAART in the HIV/TB group and the HIV group were 8.91% +/- 4.82% vs. 2.63% +/- 2.34% respectively (P < 0.01). Treg were decreased more rapidly after HAART in the former than the latter.

CONCLUSIONSTB and HAART both had an effect on the Th17/Treg ratio of HIV/ TB co-infected patients, which can cause increased Th17 expression, the later plays a pro-inflammatory role. TB and HAART can decrease Treg expression and enhance anti-inflammation response. The fact that Th17/ Treg disorder are more likely to exist in patients with HIV/TB co-infection after HAART for one month suggests a potential role for Th17/Treg imbalance leading to tuberculosis-associated immune reconstitution inflammatory syndrome during patients receiving HAART period.

Adult ; Antiretroviral Therapy, Highly Active ; Coinfection ; drug therapy ; immunology ; virology ; Female ; HIV Infections ; drug therapy ; immunology ; virology ; Humans ; Male ; T-Lymphocytes, Regulatory ; drug effects ; immunology ; Th17 Cells ; drug effects ; immunology ; Tuberculosis ; immunology ; virology