Objective:To evaluate the anti-HIV-1 activity of two new nonnucleoside reverse transcriptase inhibitors (NNRTIs), JB25 and JB26, in combination with 3 approved drugs (AZT, EFV, SQV)in vitro.Methods:The serially diluted 10 concentrations of JB25 and JB26 were combined with 7 serially diluted AZT, EFV and SQV respectively.The combination was added to 384 cell culture plates and then cocultured with HIV-1 ⅢB infected MT-2 cells for 3 days. Finally, the HIV-1 production was determined by measuring the expression of reporter genes of TZM bl cells. The data were analyzed by MacSynergy Ⅱ software.Results:The average capacity of synergism/antagonism of JB25 with AZT, EFV and SQV was 244.45/-5.05(nmol/L)~2%, 119.58/-65.93 (nmol/L)~2% and 145.83/-0.32 (nmol/L)~2% respectively;the average capacity of synergism/antagonism of JB26 with AZT, EFV and SQV was 398.90/0(nmol/L)~2%, 103.62/-0.49(nmol/L)~2% and 138.473/-0.27 (nmol/L)~2% respectively. Conclusion:Two new NNRTIs JB25 and JB26 develop synergism when combined with 3 approved drugs, respectively. MacSynergy Ⅱ software could evaluate the anti-HIV-1 activity of drug combination.

OBJECTIVETo compare changes of hypothalamus-pituitary-adrenal axis (HPAA) in different rat models of Gan stagnation (GS), Pi deficiency (PD), Gan stagnation Pi deficiency (GSPD) syndromes, and to observe interventional effect of Chaishu Sijun Decoction (CSD, capable of soothing Gan-qi invigorating Pi) on them.

METHODSSeventy Wistar rats were divided into the normal control group (group 1), the GS group (group 2), the PD group (group 3), the GSPD group (group 4), the GS intervention group (group 5), the PD intervention group (group 6), and the GSPD intervention group (group 7) according to random digit table, 10 in each group. Rats in group 1 received no treatment. Rats in group 2 and 5 were modeled by chronic restraint method. Rats in group 3 and 6 were modeled by excess fatigue plus alimentary abstinence method. Rats in group 4 and 7 were modeled by chronic restraint, excess fatigue, and alimentary abstinence method. At the 2nd weekend of modeling, CSD at 2.86 g/kg was fed to rats in group 5, 6, and 7 by gastrogavage for 2 successive weeks. Equal volume of distilled water was given to rats in the rest 4 groups. On the 29th day, rats were killed, adrenal weight weighed, and adrenal index calculated. Levels of plasma and hypothalamus corticotropin-releasing hormone (CRH), plasma and pituitary adrenocorticotrophic hormone (ACTH), and plasma corticosterone (CORT) were determined using radioimmunity.

RESULTSCompared with group 1, adrenal index significantly decreased in group 2, 3, and 4 (P < 0.05). Of them, plasma and hypothalamus CRH, plasma CORT increased significantly in group 2 and 4 (P < 0.05). Besides, plasma and pituitary ACTH increased in group 4 (P < 0.05). Plasma and pituitary ACTH, as well as plasma CORT decreased significantly in group 3 (P < 0.05). Compared with group 2, 3, and 4, adrenal index increased significantly in group 5, 6, and 7 (P < 0.05). Compared with group 2, plasma CORT, hypothalamus CRH, and pituitary ACTH decreased significantly in group 5 (P < 0.05). Compared with group 3, plasma ACTH and CORT increased significantly in group 6 (P < 0.05). Compared with group 4, plasma CRH, ACTH, CORT, hypothalamus CRH, and pituitary ACTH decreased in group 7 (P < 0.05).

CONCLUSIONSThe function of HPA .axis was damaged to varying degrees in rats of the three models in this experiment. Hyperactivity of HPA axis existed in GS syndrome and GSPD syndrome. Impairment of feedback regulation in hypothalamus and pituitary was accompanied in GSPD syndrome. Hypofunction of HPA axis existed in PDS. CSD, capable of soothing Gan-qi invigorating'Pi, showed improvement on disarranged HPAA, but with optimal effect on GSPD syndrome. CSD had higher correlation with GSPD syndrome.

Adrenocorticotropic Hormone ; metabolism ; Animals ; Corticosterone ; Corticotropin-Releasing Hormone ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hypothalamo-Hypophyseal System ; metabolism ; Hypothalamus ; metabolism ; Medicine, Chinese Traditional ; Models, Animal ; Pituitary Gland ; metabolism ; Pituitary-Adrenal System ; metabolism ; Rats ; Rats, Wistar

AIMTo test the stability of marine polysaccharide drug sulfated polyguluronic acid ester.

METHODSFour methods including high performance gel chromatography (HPGC), poly-acrylamide gel electrophoresis (PAGE), UV scan of absorbance between 200 and 800 nm and gelatin nephelometry were established. Samples were tested in high temperature, high humidity, strong light and accelerated test conditions. The methods were used to test the changes of the parameters including molecular weight, molecular weight distribution, absorbance between 200 and 800 nm, free sulfate, with which we could estimate the stability of sulfated polyguluronic acid ester could be estimated.

RESULTSThe four methods were suitable to test the stability of sulfated polyguluronic acid ester and the sample were stable in the conditions as before except in high temperature.

CONCLUSIONSulfated polyguluronic acid ester has good stability.

Chromatography, Gel ; methods ; Drug Stability ; Electrophoresis, Polyacrylamide Gel ; methods ; Molecular Weight ; Polysaccharides, Bacterial ; chemistry ; Spectrophotometry, Ultraviolet ; methods ; Temperature

AIMTo search for colchicine derivatives which have high efficacy and low toxicity.

METHODSColchicine was firstly converted into thiocolchicine, and then it was hydrolyzed to get 7-(N-deacetylthiocolchicine). At last, 7-(N-deacetylthiocolchicine) was amidated to get the target compounds. The chemical structure of these new derivatives was confirmed with 1H NMR, IR, MS, and HR-MS. The cytotoxicity of the compounds was tested by MTT assay. Their in vivo antitumor activity was evaluated against mice tumor H22 and U14.

RESULTSTwelve thiocolchicine derivatives are new compounds.

CONCLUSIONIn vitro antitumor activity has showed that some of these thiocolchicines possessed cytotoxic activity superior to colchicine. However, in vivo antitumor activity indicated that these derivatives have poor efficacy in mice.

Animals ; Antineoplastic Agents, Phytogenic ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Survival ; drug effects ; Colchicine ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Humans ; Inhibitory Concentration 50 ; Liver Neoplasms, Experimental ; pathology ; prevention & control ; Male ; Mice ; Mice, Inbred ICR ; Models, Chemical ; Molecular Structure ; Neoplasm Transplantation ; Prostatic Neoplasms ; pathology ; prevention & control ; Structure-Activity Relationship

OBJECTIVETo observe the changes in activities of superoxide dismutase (SOD) and glutathione peroxide (GSH-Px) in the induced sputum of silicosis patients, and to investigate the roles of SOD and GSH-Px in the development and progression of silicosis and the significance of measuring activities of SOD and GSH-Px in induced sputum among silicosis patients.

METHODSFifty hotel attendants were chosen as control group, 50 workers with more than one year of silica dust exposure as dust exposure group, 32 silica dust-exposed workers as observation subject group, and 52 silicosis patients as silicosis group. The activities of SOD and GSH-Px in their induced sputum were measured by enzyme-linked immunosorbent assay.

RESULTSCompared with the control group, the observation subject group and silicosis group had significantly decreased SOD activity (68.16 ± 30.17 and 66.38 ± 47.32 U/ml vs 75.81 ± 11.92 U/ml, P < 0.05); compared with the dust exposure group, the silicosis group had significantly decreased SOD activity (66.38 ± 47.32 U/ml vs 70.12 ± 14.31 U/ml, P < 0.05). Compared with the control group and dust exposure group, the observation subject group and silicosis group had significantly increased GSH-Px activity (268.21 ± 15.45 and 279.34 ± 29.26 U/ml vs 224.22 ± 12.64 and 236.41 ± 14.54 U/ml, P < 0.05 or P < 0.01).

CONCLUSIONThe SOD activity in dust exposure group and silicosis group decreased, but there were no significant differences between patients with different stages of silicosis. The GSH-Px activity in dust exposure group and silicosis group was significantly higher than that in control group, and there were significant differences between patients with different stages of silicosis. These suggest that the imbalance of oxidative/antioxidant systems is associated with the development and progression of silicosis.

Adult ; Case-Control Studies ; Glutathione Peroxidase ; metabolism ; Humans ; Middle Aged ; Silicosis ; enzymology ; Sputum ; enzymology ; Superoxide Dismutase ; metabolism

The gene GeDRP1E encoding dynamin-related protein 1E in Gastrodia elata was cloned by specific primers which were designed based on the transcriptome data of G. elata. Bioinformatics analysis on GeDRP1E gene was carried out by using ExPASy, ClustalW, MEGA, etc. Positive transgenic Arabidopsis plant and potato minituber were obtained with the genetic transformation system of Arabidopsis and potato. The plant height and seed setting rate of transgenic Arabidopsis, and agronomic characters, such as size, weight and starch content of potato minituber of transgenic potato were tested and analyzed. And GeDRP1E gene function was preliminarily investigated. The results showed that the open reading frame of GeDRP1E gene was 1 899 bp in length and 632 amino acids residues were encoded, with a relative molecular weight of 69.90 kDa and a molecular formula of C₃₀₇₉H₄₉₇₃N₈₈₃O₉₃₃S₁₉. It was predicted that the theoretical isoelectric point was 7.27, the instability coefficient was 43.34, and the average hydrophilicity index was -0.259, which was indicative of an unstable hydrophilic protein. GeDRP1E has no transmembrane structure and signal peptide, and was localized in the cytoplasm. The phylogenetic tree showed that GeDRP1E was highly homologous with DRP1E proteins of other plant species, among which GeDRP1E had the highest homology with DcDRP1E (XP_020689662.1) in Dendrobium candidum, reaching 90.05%. GeDRP1E plant expression vector pCambia1300-35Spro-GeDRP1E was constructed by double digests, and Arabidopsis complementary mutant and potato overexpression strain of GeDRP1E gene were obtained by Agrobacterium-mediated gene transformation. Compared with the Arabidopsis AtDRP1E mutant, the height and seed setting rate of the GeDRP1E complementation mutant were rescued. The minituber of GeDRP1E overexpression potato had larger size, heavier weight and higher starch content, comparing to wild-type potato. It was preliminarily induced that GeDRP1E was involved in mitochondrial morphology regulation, which related to the growth and development of Arabidopsis plants and potato miniature. The research results laid a foundation for further elucidating the molecular mechanisms underlying the growth and development of G. elata tuber development.

Angiotensin II Type 1 Receptor Blockers ; administration & dosage ; Animals ; Coronary Angiography ; Coronary Restenosis ; prevention & control ; Coronary Vessels ; pathology ; Drug Delivery Systems ; Immunohistochemistry ; Rabbits ; Receptor, Angiotensin, Type 1 ; analysis ; Receptor, Angiotensin, Type 2 ; analysis ; genetics ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Stents ; Tetrazoles ; administration & dosage ; Valine ; administration & dosage ; analogs & derivatives ; Valsartan

BACKGROUNDThe decrease of surfactant protein (SP) secreted by the alveolar type II cell is one of the important causes of limiting air of pulmonary emphysema. However, the SP-A gene and protein changes in this disease are rarely studied. This study was undertaken to investigate alterations in SP-A gene activity and protein, and to explore their roles in the pathogenesis of emphysematous changes.

METHODSTwenty Wistar rats were divided randomly into a normal control group (n = 10) and a cigarette smoking (CS) + lipopolysaccharide (LPS) group (n = 10). Ultra-structural changes were observed under an electron microscope. The number of cells positive for SP-A was measured by immunohistochemistry. The mRNA expression and protein level of SP-A in the lung tissues were determined by quantitative polymerase chain reaction (qPCR) and Western blot separately. The protein level of SP-A in lavage fluid was determined by Western blot.

RESULTSThe number of cells positive for SP-A of the CS + LPS group (0.35 +/- 0.03) was lower than that of the blank control group (0.72 +/- 0.06, P < 0.05). The level of SP-A in the lung tissues of rats in the CS + LPS group (0.2765 +/- 0.0890) was lower than that in the blank control group (0.6875 +/- 0.1578, P < 0.05). The level of SP-A in the lavage fluid of rats in the CS + LPS group (0.8567 +/- 0.1458) was lower than that in the blank control group (1.3541 +/- 0.2475, P < 0.05). The lung tissues of rats in the CS + LPS group showed an approximate increase (0.4-fold) in SP-A mRNA levels relative to beta-actin mRNA (P < 0.05).

CONCLUSIONSThe changes of SP-A may be related to emphysematous changes in the lung. And cigarette smoke and LPS alter lung SP-A gene activity and protein homeostasis.

Animals ; Blotting, Western ; Emphysema ; metabolism ; pathology ; Homeostasis ; Immunohistochemistry ; Male ; Microscopy, Electron ; Polymerase Chain Reaction ; Pulmonary Surfactant-Associated Protein A ; analysis ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar

OBJECTIVETo investigate the immunologic mechanism of acupuncture at Baihui (GV 20) and Zusanli (ST 36) for treatment of depression.

METHODSEighty-four cases of depression patients were randomly divided into an electroacupuncture observation group (group A), an electroacupuncture control group (group B) and a medication control group (group C), 28 cases in each group. Baihui (GV 20) and Zusanli (ST 36) were used in the group A; Taichong (LR 3), Sanyinjiao (SP 6), Neiguan (PC 6) and Shenmen (HT 7) were used in the group B; and the group C was treated with oral administration of Fluoxetine. The scores of Hamilton Depression Scale (HAMD) were tested and the level of serum interleukin 1 (IL-1 beta), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-alpha) were measured by Enzyme Linked Immunosorbent Assay (ELISA) before and after treatment.

RESULTSTheir scores of HAMD were obviously decreased after treatment in three groups (all P<0.01). In the group A, 2 cases were cured, 19 cases were markedly effective, 5 cases were effective and 2 cases were ineffective. In the group B, 16 cases were markedly effective, 12 cases were effective. While in the group C, 1 case was cured, 17 cases were markedly effective, 7 cases were effective and 3 cases were ineffective. The grade distribution of clinical effect showed that the effect of group A was better (P<0.05). The levels of serum IL-1 beta and IL-6 in the three groups were obviously decreased after treatment (P<0.05, P<0.01), while there was no significant difference between the level of serum TNF-alpha before and after treatment.

CONCLUSIONThe effect of electroacupuncture observation group is superior to those of electroacupuncture control group and medication control group. All of these three methods can clear the inflammatory cytokines such as IL-1 beta and IL-6 away and improve the symptoms of depression.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Cytokines ; blood ; Depression ; blood ; immunology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo explore the effects of mitogen activated protein kinase (MAPK) and extracellular signal-regulated kinases (ERK) on kidney injury in female BALB/c mice exposed to cadmium.

METHODTwenty-one female BALB/c mice were randomly divided into 3 groups, i.e. control group, low Cd exposure group (2.5 µmol/kg) and high Cd exposure group (10 µmol/kg) were exposed to normal saline, 2.5, 10 µmol/kg Cd, respectively, 3 times a week for 14 weeks. The kidney slice were stained by HE, PAS and Masson staining to observe the morphological changes. The expression levels of pERK, ERK, pp38, p38, pJNK and JNK proteins in kidneys were tested by Western blot assay.

RESULTSThe ratios of pERK/ERK, pp38/p38, pJNK/JNK in high Cd group were higher than those in the control group (P < 0.05). The ratio of pERK/ERK in low Cd group was higher than control group (P < 0.05). The expression levels of bcl-2, bax proteins and the ratio of bcl-2 to bax in Cd exposure groups decreased significantly, as compared with the control group (P < 0.05). The impairment of renal glomeruli and tubules were observed in HE, PAS and Masson staining slices of kidneys in mice exposed to Cd.

CONCLUSIONCdCl2 may induced renal injury by affecting the expression levels of MAPK.

Animals ; Apoptosis ; drug effects ; Cadmium ; toxicity ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Female ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Kidney ; metabolism ; pathology ; MAP Kinase Signaling System ; Mice ; Mice, Inbred BALB C ; Mitogen-Activated Protein Kinases ; metabolism ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; metabolism