OBJECTIVETo investigate the effect of four traditional Chinese medicines distributed along lung meridian, namely Ephedrae Hebra, Zingiberis Rhizoma, Scutellariae Radix and Mori Cortex, on TLR2 and NF-κB expressions in mice with lung heat syndrome, in order to study the intervention effect of the four traditional Chinese medicines (TCMs) on the lung heat syndrome.

METHODOne hundred KM mice were randomly divided into the normal control group, the model group, the Ephedrae Hebra group, the Zingiberis Rhizoma group, the Scutellariae Radix group and the Mori Cortex group (20, 10 g x kg(-1)), nasally dripped with streptococcus pneumoniae to establish the mouse lung heat syndrome model, and then administered with different TCMs. The expressions of TLR2, NF-κB p65 proteins in lung tissues were analyzed by the immunohistochemical method. The expressions of TLR2, NF-κB p65 mRNA were measured by real time PCR.

RESULTCompared with the normal control group, the expressions of TLR2 and NF-κB p65 proteins in lung tissues in the model group were higher (P < 0.01), and the expressions of TLR2 and NF-κB p65 mRNA in lung tissues were up-regulated (P < 0.01 or P < 0.05). Compared with the model group, Ephedrae Hebra high and low dose groups, the Zingiberis Rhizoma low dose group and the Scutellariae Radix high dose group showed decreased expression of TLR2 protein (P < 0.05 or P < 0.01); Ephedrae Hebra high and low dose groups, the Zingiberis Rhizoma low dose group, Scutellariae Radix high and low dose groups and Mori Cortex high and low dose groups showed reduced expression of NF-κB p65 protein (P < 0.05 or P < 0.01). Ephedrae Hebra high and low dose groups, Zingiberis Rhizoma high and low dose groups, Scutellariae Radix high dose group and Mori Cortex high dose group showed down-regulated expression of TLR2 mRNA (P < 0.01 or P < 0.05).

CONCLUSIONEphedrae Hebra, Zingiberis Rhizoma, Scutellariae Radix and Mori Cortex can induce the TLR2/NF-κB inflammatory signal pathways by down-regulating the expressions of TLR2 and NF-κB p65 in protein and mRNA, so as to alleviate the lung tissue injury in mice with lung heat syndrome.

Animals ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; classification ; pharmacology ; Female ; Gene Expression ; drug effects ; Immunohistochemistry ; Lung ; drug effects ; metabolism ; microbiology ; Male ; Medicine, Chinese Traditional ; methods ; Meridians ; Mice ; Phytotherapy ; methods ; Plants, Medicinal ; chemistry ; classification ; Pneumonia, Pneumococcal ; drug therapy ; genetics ; metabolism ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction ; Scutellaria baicalensis ; chemistry ; Species Specificity ; Syndrome ; Toll-Like Receptor 2 ; genetics ; metabolism ; Transcription Factor RelA ; genetics ; metabolism

Objective To analyze the factors affecting the efficacy of postoperative radiotherapy (PORT) in node-positive non-small cell lung cancer (NSCLC). Methods 480 patients with stage N_1-N_2 NSCLC after radical surgery were retrospectively reviewed. Of them, 267 patients received adjuvant chemotherapy and 121 received PORT. All patients were grouped based on the N stage, tumor size and lymph node positive ratio (the percentage of positive lymph nodes from the detected lymph nodes, LNPR). Group 1 included patients with tumor size ≤3 cm and LNPR ≤33%, group 2 was tumor size > 3 cm or LNPR > 33%, and group 3 was tumor size > 3 cm and LNPR > 33%. The endpoints were the local recurrence free survival (LRFS) and overall survival (OS). Kaplan-Meier method and Cox's proportional hazards regression model were used for the statistic analyses. Results PORT improved the overall survival only in patients with N_2 disease. Both tumor size and LNPR significantly influenced the efficacy of PORT. The 5-year LRFS for patients with vs. without PORT in the group 1, 2 and 3 were 55% vs. 60% (χ~2 = 0.03,P-0.869), 42% vs. 50% (χ~2 =0.31,P=0.547),and 62% vs. 52% (χ~2=4.25,P=0.036), respectively;and the corresponding OS were 22% vs. 50% (χ~2 = 1.65 ,P =0. 199), 26% vs. 22% (χ~2= 0. 13,P=0.786) and 42% vs. 16% (χ~2= 15.33,P=0.000), respectively. Conclusions Tumor size and LNPR significantly impact the efficacy of PORT . For patients with stage N_2 NSCLC , PORT could improve local recurrence free survival and overall survival when tumor size > 3 cm and LNPR >33%.

Objective With awareness, attitudes, and rice-intake behavior of Kaschin-Beck disease (KBD) and the analysis of the factors that influence KBD related rice-intake behaviors among resident's in Aba,this research could provide evidences for KBD-Control, and benefit the policy development related to KBD-Control.Methods Villages were chosen by proportional stratified random sampling from KBD monitoring villages among agriculture areas, pastoral areas, and farming & pastoral areas in Aba, Sichuan, in July 2009. Interview questionnaire of household survey, designed by research associates of this project, was used in this research for residents in endemic area of KBD in Aba. The questionnaire covered demographic and socio-economic characteristics, KBD knowledge and diet habits. Multi-level Variance Component Analysis was used to explore factors which would influence the KBD related rice-intake behaviors. Results A total of 1029 permanent residents were recruited in this research, among which the detection rate of KBD was 48.01％ (482/1004). Most of the patients lived in farming & pastoral areas(84.44％, 407/482). Pastoral residents had the least knowledge of KashinBeck disease, and the composition ratios ofGeneral andGood were 15.87％ (33/208)and 3.36％ (7/208),respectively. Still, people who were willing to have rice as staple food were 93.13％(935/1004). It indicated that only (50.40 ± 23.68)％ on average, of research subjects had the life style of rice intake. Ethnic, work status,language situation and attitudes to rice intake were influencing factors for rice-intake behavior. Conclusions The percentage of rice intake in Aba KBD epidemic areas is low. And to prevent KBD, the advocacy actions should be targeted at ethnic, work status, language situation, and attitudes to rice intake.

AlM:To discuss the protective effect ofα-mangostin on retinal light damage in mice.METHODS:Totally 30 Balb/c mice, aged 6~8wk, were randomly divided into the control group, light-exposure group and α-mangostin group. Every group contained 10 mice. Mice of α-mangostin group were treated with alpha-mangostin at the dose of 30mg/( kg · d ) body weight by intragastric administration daily for 7d, and then exposed to white light at the 5th d. The light-exposure group and α-mangostin group were exposed to 5 000 ± 200lx white light-emmiting diodes (LEDs) for continuously 1h to establish the mice model of retinal light damage. Flash -electroretinograme was recorded 72h after light exposure. The changes in retinal morphology of mice were observed by light microscopy. Retinas were extracted to detect the malondialdhyde ( MDA ) content change of the retinal homogenate.RESULTS: Flash-electroretinogram ( F-ERG ) showed that retinal dysfunction was less severe in α-mangostin group than in light-exposure group ( P<0. 05 ). Light microscopy test showed that retina structural damage was less severe in α-mangostin group than in light-exposure group (P<0. 05). The level of MDA in retinal tissue of α-mangostin group was significantly lower when compared with light-exposure group (P<0. 05).CONCLUSlON: α-mangostin inhibits lipid peroxidation induced by light damage and protect retina against light damage.

Objective To study the clinical char acteristics and outcome of two lymphoma patients mimicking Beh?et's disease. Methods Lymphoma was diagnosed in two patients mimicking Beh?et's disease referred to our Department in 2015. A search on published similar cases in Chinese database and the Pubmed was also performed and then analyzed. Results Eight patients were indentified in this pooled analysis, six of which were non-Hodgkin lymphoma (NHL). All of the eight cases presented with cutaneous lesion, seven cases with fever, seven cases with oral ulceration and six cases with orogenital ulceration, respectively. Ocular involvement was present in four of the eight cases, two were with a positive pathergy test. Among feverish patients, six were moderate or high fever, four were high fever, one was low-grade fever. Neutropenia was found in four patients, and lymphocytoponia in four of five patients with detailed data. All patients fulfilled the 2014 International Criteria for Beh?et's Disease (ICBD) with an average score of (5.8 ±1.5), ranging from 4 to 8. Survival period ranged from one month to 36 months, with an average of 8 months. Conclusion For patients diagnosed as Beh?et's disease are finally diagnosed as lymphoma. For patients with Beh?et's disease present-ation but also present with mediate to high fever, atypical deepseated ulcer, neutropenia or lymphocytoponia, malignancy especially lymphoma should be investigated.

To establish single- and double-transfected transgenic cells stably expressing hMATE1, hMATE1 cDNA was cloned by RT-PCR from human cryopreserved kidney tissue, and subcloned into pcDNA3.1(+) plasmid by virtue of both HindIII and Kpn I restriction enzyme sites. Subsequently, the recombined pcDNA3.1(+)- hMATE1 plasmid was transfected into MDCK, MDCK-hOCT1 or MDCK-hOCT2 cells using Lipofectamine 2000 Reagent. After a 14-day-cultivation with hygromycin B at the concentration of 400 µg · mL(-1), all clones were screened with DAPI and MPP+ as substrates to identify the best candidate. The mRNA content of hMATE1, the cellular accumulation of metformin with or without cimetidine as inhibitor, or transportation of cimetidine was further valuated. The results showed that all of the three cell models over expressed hMATE1 mRNA. The cellular accumulation of metformin in MDCK-hMATE1 was 17.6 folds of the control cell, which was significantly inhibited by 100 µmol · L(-1) cimetidine. The transcellular transport parameter net efflux ratios of cimetidine across MDCK-hOCT1/hMATE1 and MDCK-hOCT2/hMATE1 monolayer were 17.5 and 3.65, respectively. In conclusion, cell models with good hMATE1 function have been established successfully, which can be applied to study the drug transport or drug-drug interaction involving hMATE1 alone or together with hOCT1/2 in vitro.
Animals ; Biological Transport ; Cimetidine ; pharmacology ; DNA, Complementary ; Dogs ; Drug Interactions ; Humans ; Madin Darby Canine Kidney Cells ; Metformin ; pharmacology ; Organic Cation Transport Proteins ; genetics ; metabolism ; Transfection

In this study, based on the transcriptome data, we cloned the full-length cDNAs of TwAACT gene from Tripterygium wilfordii suspension cells, and then analyzed the bioinformation of the sequence, detected the genetic differential expression after being induced by methyl jasmonate (MeJA) by RT-PCR. The full-length cDNA of the TwAACT was 1 704 bp containing a 1 218 bp open reading frame (ORF) encoding a polypeptide of 405 amino acids (GeneBank accession No. KP297934). The deduced isoelectric point (pI) was 6.10, a calculated molecular weight was about 41.20 kDa, and online prediction showed that TwAACT had two catalytic active sites. After the induction of MeJA, the relative expression level of TwAACT increased rapidly. The expression level of TwAACT was highest at 24 h. TwAACT was cloned firstly, that laid the foundation for identifying thegene and illustrating thebiosynthesis mechanism and its synthetic biology.
Acetyl-CoA C-Acetyltransferase ; chemistry ; genetics ; metabolism ; Amino Acid Sequence ; Cloning, Molecular ; Gene Expression Regulation, Plant ; Models, Molecular ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Sequence Alignment ; Tripterygium ; chemistry ; classification ; enzymology ; genetics

Objective:To stablish of chemiluminescent Southern blot detection system for examining HBV DNA replication intermediates in HepG2.2.15,and analyse the inhibition of HBV replication with three kind of drugs with different targets.Methods:The HBV DNA replication intermediates were extracted and analyzed by Southern blot with HBV probe,which(pTHBV1047) was labelling with digoxigenin.The results of the hybridization were detected by chemiluminescent,and the condition of hybridization was optimized.After treated with lamivudine,Bay41-4109,?-Galcer in different concentration,the HBV DNA from the HepG2215 cells were detected with the system.Results:the sensitivity of the system was 1pg of pTHBV1047,and HBV specific positive signals was detected with the DNA from HepG2.2.15.The three kinds of drugs can inhibit the HBV replication obviously with chemiluminescent Southern blot detection system,the IC50 were 1.53?mol/L,0.41?mol/L,0.01?mol/L.Conclusion:The HBV replication intermediates from the cell of HepG2.2.15 can reflect the antiviral effect accurately with different targets drugs and this mothod would be used in the study of Chinese-midicine.

The present study was to investigate the role of dopamine D1 receptors and its relationship with glutamate, N-methyl-D-aspartic acid (NMDA) receptor and α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor in depression induced by chronic unpredictable mild stress (CUMS). CUMS-induced depression model was established in Sprague-Dawley rats, and intrahippocampal microinjections of D1 dopamine receptor agonist SKF38393, non-competitive NMDA receptor antagonist MK-801 and AMPA receptor antagonist NBQX were respectively adopted by rat brain stereotaxic coordinates. The behavioral observations were conducted by measurement of weight changes, sucrose preference test, open-field test and tail suspension test. The concentration of glutamic acid and the expression of its receptors' subunits were detected by HPLC and Western blot, respectively. The results showed that, compared with control group, CUMS rats showed depression-like behavioral changes, higher concentration of glutamic acid, lower expressions of NMDA receptor (NR1) and AMPA receptor (GluR2/3) in hippocampus. Pretreatment with injection of SKF38393 could rescue such depression effect of CUMS, decrease the concentration of glutamic acid, and increase the expressions of NMDA receptor (NR1), AMPA receptor (GluR2/3) in hippocampus. Pretreatment with MK-801 could enhance the antidepressant effect of SKF38393, while NBQX weakened. These results suggest that agonists of D1 dopamine receptor could reduce the concentration of glutamic acid in hippocampus, and its antidepressant effect may be mediated by AMPA receptor partially.
2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-benzazepine ; pharmacology ; Animals ; Depression ; etiology ; physiopathology ; Dizocilpine Maleate ; pharmacology ; Excitatory Amino Acid Antagonists ; Glutamates ; metabolism ; Hippocampus ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, AMPA ; metabolism ; Receptors, Dopamine D1 ; agonists ; physiology ; Stress, Physiological ; physiology