C6-structural optimizations of 2-aryl-1H-pyrazole-S-DABOs: From anti-HIV to anti-DENV activity.

Both HIV and DENV are serious threats to human life, health and social economy today. So far, no vaccine for either HIV or DENV has been developed successfully. The research on anti-HIV or DENV drugs is still of great significance. In this study we developed a series of novel 2-Aryl-1H-pyrazole-S-DABOs with C6-strucutral optimizations as potent NNRTIs, among which, 8 compounds had low cytotoxicity and EC50 values in the range of 0.0508 âˆ¼ 0.0966 µM, and their selectivity index was SI > 1415 âˆ¼ 3940. In particular, two compounds 4a and 4b were identified to have good inhibitory effects on DENV of four serotypes. The EC50 of compound 4a and 4b against DENV-II (13.2 µM and 9.23 µM, respectively) were better than that of the positive control ribavirin (EC50 = 40.78 µM). In addition, the effect of C-6 substituents on the anti-HIV or anti-DENV activity of these compounds was also discussed.

The New NNRTI ACC007 Combined with Lamivudine and Tenofovir Disoproxil Fumarate Show Synergy Anti-HIV Activity In Vitro.

BACKGROUND: Acquired immunodeficiency syndrome can hardly be cured currently and people with human immunodeficiency virus (HIV) need lifelong treatment that may result in the emergence of drug resistance which leads to failed treatment. Thus, the development of new anti- HIV drugs and new treatment regimens are necessary. OBJECTIVE: The aim of this study is to analyze the combined anti-HIV activity of tenofovir disoproxil fumarate, lamivudine and ACC007, a new non-nucleoside reverse transcriptase inhibitor. METHODS: The antiviral activity of tenofovir disoproxil fumarate, lamivudine and ACC007 alone or in combination against different HIV-1 strains was determined by the detection of HIV-1 p24 level through enzyme-linked immunosorbent assay. RESULT: ACC007 showed EC50 of nanomolar range (from 3.03 nM to 252.59 nM) against all HIV-1 strains used in this study except the HIV-1A17, with EC50 of 1.57 µM. The combined antiviral activity of ACC007, lamivudine and tenofovir disoproxil fumarate showed synergy antiviral activity against all HIV-1 strains used in this study. The three-drug combination showed moderate synergism against HIV-1A17, HIV-14755-5, HIV-1K103N and HIV-1V106M, with a combination index value ranging from 0.71 to 0.87, and showed synergism against the other HIV-1 strains with combination index value from 0.35 to 0.67. The combination with ACC007 significantly increases the dose reduction index value of lamivudine and tenofovir disoproxil fumarate, compared with two-drug combination. CONCLUSION: ACC007 exhibits potent antiviral activity alone or with 3TC and TDF, and exerts synergistic effect against all HIV strains used in our investigation in vitro.

[Effect of Electroacupuncture Intervention on Behavior Changes and Levels of Hippocampal Transforming Growth Factor beta 3 and Basic Fibroblast Growth Factor Proteins in Depression Rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on behavior changes and the adundance levels of transforming growth factor beta 3 (TGF-beta 3) and basic fibroblast growth factor (bFGF) proteins in the hippocampus of rats with chronic unpredictable mild stress (CUMS)-induced depression, so as to explore its mechanisms underlying improvement of depression. METHODS: Forty Sprague-Dawley rats were randomly divided into the following groups: control, model, EA, and medication (Fluoxetine), n = 10 in each group. The depression model was established by CUMS combined with solitary raising for 28 days. EA (2 Hz, 0.6 mA) was applied to "Baihui" (GV 20) and "Yintang" (GV 29) for 20 mm, once daily before CUMS every day. The rats of the medication group were given with Fluoxetine (10 mg/kg, 5 mL/kg) before CUMS every day. The behavioral changes (crossing and rearing locomotion) were detected by using open field tests. The expression levels of TGF-beta 3 and bFGF proteins of the bilateral hippocampus tissues were detected using biotin label-based antibody protein chips. Results Compared to the control group, the crossed grid-square numbers and rearing times were significantly decreased in the model group (P<0.01). Following EA and medication interventions, the CUMS induced decreases of the crossed grid-square number and rearing times were notably reversed in both EA and medication groups (P<0.01), suggesting an amelioration of depression after the intervention. The relative expression level of hippocampal TGF-beta 3 was down-regulated (fold change = 0.48, vs. the control group) and that of bFGF up-regulated (fold change= 1.36, vs the control group) in the model group. In both the EA and medication groups, the down-regulated TGF-beta 3 expression and the up-regulated bFGF protein expression were suppressed (TGF-beta 3: fold change = 1.61, 1.6 and bFGF: fold change = 0.61, 0.45, vs. the model group respectively). CONCLUSION: EA can improve the depression-like state in depression rats which may be associated with its effect in up-regulating hippocampal TGF-beta 3 protein level and down-regulating bFGF protein expression via promoting neurogenesis.

[Effect of Acupuncture Intervention on c-jun N-terminal Kinase Signaling in the Hippocampus in Rats with Forced Swimming Stress].

OBJECTIVE: To observe the effect of acupuncture on c-jun N-terminal Kinase (JNK) signaling in the hippocampus in rats with forced-swimming stress, so as to reveal its underlying mechanism in relieving depression-like motor response. METHODS: Forty-eight Sprague-Dawley rats were randomly divided into 8 groups as control, control + JNK inhibitor (SP 600125) , model, model + SP 600125, acupuncture, acupuncture + SP 600125, Fluoxetine (an anti-depressant) , and Fluoxetine + SP 600125 (n = 6 in each group). The depression-like behavior (immobility) model was established by forcing the rat to swim in a glass-cylinder and solitary raise. Acupuncture stimulation was applied to "Baihui" (GV-20) and "Yintang" (GV 29) for 20 min before forced swimming and once again 24 h later.. The rats of the Fluoxetine and Fluoxetine+ SP 600125 groups were treated by intragastric administration of fluoxetine 10 mL (1.8 mg)/kg before forced swimming and once again 24 h thereafter. The rats of the model + SP 600125 and acupuncture + SP 600125 groups were treated by intraperitoneal injection of SP 600125 (10 mg/kg) 90 min before forced swimming and 30 min before acupuncture intervention, respectively. The immobility duration of rats in the water glass-cylinder was used to assess their depression-like behavior response. The expression levels of protein kinase kinase 4 (MKK 4), MKK 7, JNK, and phosphorylated JNK (p-JNK) in the hippocampus were detected by Western blot. RESULTS: Compared to the control group, the duration of immobility, and the expression levels of hippocampal MKK 4, MKK 7, and p-JNK proteins were significantly increased in the model group (P < 0.01). While in comparison with the model group, the duration of immobility in the model + SP 600125, acupuncture, acupuncture + SP 600125, Fluoxetine and Fluoxetine + SP 600125 groups, the expression levels of hippocampal MKK 4 and MKK 7 proteins in the Fluoxetine + SP 600125 group, and those of p-JNK protein in the acupuncture, acupuncture + SP 600125, model + SP 600125, Fluoxetine and Fluoxetine + SP 600125 groups were considerably decreased (P < 0.05, P < 0.01). No significant differences were found between the control and control + SP 600125 groups and among the model + SP 600125, acupuncture, acupuncture + SP 600125, Fluoxetine and Fluoxetine + SP 600125 groups in the duration of immobility (P > 0.05), and in the expression level of p-JNK protein (P > 0.05). No significant changes were found in the expression levels of JNK among the 8 groups (P > 0.05). CONCLUSION: Acupuncture stimulation of GV 20 and GV 29 is effective in relieving depression-like motor response in forced-swimming stress rats, which may be closely associated with its effects in down-regulating the expression of hippocampal p-JNK protein.

Generation of the CF3 radical from trifluoromethylsulfonium triflate and its trifluoromethylation of styrenes.

The CF(3) radical was generated from the reaction of S-(trifluoromethyl)diphenylsulfonium triflate with Na(2)S(2)O(4) or HOCH(2)SO(2)Na under suitable conditions without further reduction. Based on this, a method for the synthesis of α-trifluoromethylated ketones has been successfully developed.

Detection of HPV types and neutralizing antibodies in women with genital warts in Tianjin City, China.

The serum samples and corresponding cervical swabs were collected from 50 women with genital warts from Tianjin city, China. The neutralizing antibodies against HPV-16, -18, -58, -45, -6 and -11 in serum samples were tested by using pseudovirus-based neutralization assays and HPV DNAs in cervical swabs were also tested by using a typing kit that can detect 21 types of HPV. The results revealed that 36% (18/50) of sera were positive for type-specific neutralizing antibodies with a titer range of 160-2560, of which 22%(11/50), 12%(6/50), 10%(5/50), 4%(2/50), 4%(2/50) and 2%(1/50) were against HPVs -6, -16, -18, -58, -45 and -11, respectively. Additionally, 60% (30/50) of samples were HPV DNA-positive, in which the most common types detected were HPV-68(18%), HPV-16(14%), HPV-58(12%), HPV-33(8%) and HPV-6, HPV-11, HPV-18 and HPV-52 (6% each). The concordance between HPV DNA and corresponding neutralizing antibodies was 56% (28/50) with a significant difference (P<0.05). The full-length sequences of five HPV types (HPV -42, -52, -53, -58 and -68) were determined and exhibited 98%-100% identities with their reported genomes. The present data may have utility for investigating the natural history of HPV infection and promote the development of HPV vaccines.

[The impact of the HIV-1 envelope (Env) mutation on its assembly of functional pseudovirus].

To find out whether the mutations of HIV-1 Env have influence on the assembly of pseudovirus and their abilities to infect cells, site-directed mutation (A457D)was performed using cycling mutagenesis and selection of mutants with DpnI. Transformation and plasmid purification technologies were used to obtain mutated env clone. Then both the prototype and the mutant were co-transfected with pSG3(delta(env)) to 293FT cells, respectively. Single-cycle infection assay was employed to analyze the effect of the prototype and the mutant on the ability of functional pseudovirus assembly. The transient expression of both the prototype S12-42-1 and mutant S12-42M were confirmed by Western blot essay. The S/CO value was less than 1 for S12-42-1 and 6.65 for S12-42M, demonstrating the functional pseudovirus was generated only for S12-42M. So mutation on HIV-1 Env has influence on the assembly of pseudovirus and their abilities to infect cells.

[The preliminary analysis on immunogenicity of DNA vaccines against human papillomavirus 58].

In order to evaluate the immunogenicity of HPV 58 L1 DNA vaccines, five DNA vaccines had been constructed with pcDNA3.1 vector containing different L1 genes of HPV 58, which were designated as L1h, L1hDeltac, L1S, L1SM and L1wt. The protein expression of DNA vaccines in vitro was tested by Western blot. The ability of forming pseudovirus was evaluated by transfecting DNA vaccine together with pcDNA3.1-h58L2 and pcDNA3.1-GFP into 293FT cells. The neutralizing antibodies and cellular immune response produced in BALB/c mice immunized with the DNA vaccines were detected by using pseudovirus-based neutralization assay and ELISPOT respectively. The results showed that the five DNA vaccines had been successfully constructed; the level of protein expression of L1hDeltac was the highest and those for L1S and L1SM were of medium, while no expressed target protein of L1wt was detected. Only L1S could form the pseudovirus while the other four vaccines could not. L1S and L1h could induce neutralizing antibody. However, the average titer of neutralizing antibody for L1S (1:6,400) was much higher than that for L1h (1:48) and the other three vaccines could not induce neutralizing antibody. No cellular immune response for all five DNA vaccines was detectable by ELISPOT. The results indicated that DNA vaccine against HPV 58 can form pseudovirus in vitro, also can induce high level of neutralizing antibodies. This provides reference for screening HPV vaccine in future.

[Responses of Chinese pine tree ring in Shenyang suburb (Fu Mausoleum) to global temperature fluctuation].

In this paper, the correlations between the variations of Chinese pine (Pinus tabulaeformis Carr.) tree ring width in Shenyang suburb (Fu Mausoleum) and the local temperature variables, Global Surface Air Temperature Anomaly (GSATA) from 1880 to 2004, Global Land-Ocean Temperature (GLOTI) from 1880 to 2004 and North Hemisphere Temperature Anomaly (NHTA) from 1880 to 2004 were studied. Some close correlations were detected, and the local temperature variables, GSATA, GLOTI and NHTA had some similar influences on the Chinese pine tree ring width. The air temperature in last winter (December and January) and in spring (April and May) affected the growth of Chinese pine significantly (P < 0.05). There existed a 3-8-year periodicity of the variation of Chinese pine tree ring width and the GSATA, GLOTI and NHTA, and the 19.3-year and 23.2-year quasi-periodicity of the variation of Chinese pine tree ring width corresponded with the 20.8-year quasi-periodicity of GSATA, GLOTI and NHTA. This study suggested that the Chinese pine tree ring width in Shenyang Fu Mausoleum had positive correlations with global-scale temperature fluctuation, and the temperature increase in the past had a positive effect on the Chinese pine growth.

[Evaluation of procleix HIV/HCV RNA diagnostic assay].

BACKGROUND: To investigate the sensitivity and specificity of Procleix HIV/HCV RNA diagnostic assay. METHODS: HIV antibody positive or suspected positive plasmas of blood donors were collected from different provinces and detected with HIV antibody ELISA and HCV antibody ELISA. Samples positive for HIV by ELISA were confirmed by using HIV Blot. All the plasma samples were detected with Procleix HIV/HCV assay, HIV-1 discriminatory assay and HCV discriminatory assay, respectively. RESULTS: All 74 samples positive for both HIV and HCV antibody were positive and 5 samples negative for both HIV and HCV antibody were negative when detected using Procleix HIV/HCV assay; 82 of 84 supplemental HIV antibody positive samples and 6 of 12 supplemental indeterminate samples were positive for HIV RNA, and all 7 HIV antibody negative samples were negative for HIV RNA when detected by using Procleix HIV discriminatory assay. Seventy of 81 HCV antibody positive samples and 4 of 22 HCV antibody negative samples were positive for HCV RNA when detected by using Procleix HCV discriminatory assay. CONCLUSION: This reagent is more sensitive and could be used in blood screening, thereby can reduce both HIV and HCV transmission of blood in window period of HIV and HCV infection.

[Establishment of the reference panel for HIV RNA].

OBJECTIVE: To establish a national reference panel for HIV RNA diagnostic reagents. METHODS: Sera from patients with HIV infection and healthy blood donors were collected and tested for HIV and HCV antibodies and HBsAg by using ELISA. The HIV antibody positive samples with ELISA were confirmed with HIV Blot 2.2 (Genelabs). The quantitative samples for HIV RNA were calibrated with the WHO HIV RNA standard. The stability of the panel was evaluated with acceleration method. RESULTS: After screening and calibration, 8 negative samples, 8 positive samples, 3 quantitative samples, 6 sensitivity samples and 5 samples for linear analysis were composed of the national reference panel for HIV RNA. The convinced international units (IU) for the quantitative samples were obtained by seven independent calibration and the logarithm of international units for the quantitative samples (b1-b3) were less than x +/- s. The results showed that this panel may stabilize for 4 days at 4 degrees C. CONCLUSION: A national reference panel for HIV RNA reagents has been established. It may provide the basis for evaluating HIV RNA diagnostic reagents.

[Capacity of HIV enzyme immunoassay diagnostic kits to detect antibodies against different genotypes of HIV].

OBJECTIVE: To investigate the capacity of commercial HIV enzyme immunoassay (EIA) diagnostic kits to detect antibodies against different genotypes of HIV. METHODS: HIV RNA was detected with RT-PCR from samples positive for HIV antibody. The purified PCR products were sequenced directly and the genotypes of HIV from samples were analyzed. The samples for each genotype of HIV were diluted and the diluted samples were detected with different HIV EIA diagnostic kits. RESULTS: All 20 samples positive for HIV antibody were also positive for HIV RNA; 9 of 20 isolates were genotype B, 9 of them were genotype C or CRF BC, 2 of them were CRF AE. The sensitivity of different HIV EIA diagnostic kits to detect antibodies against different genotypes of HIV was not significantly different. CONCLUSION: The capacity of commercial HIV diagnostic kits to detect antibodies against different HIV genotypes may not be significantly different.