In this study, we assessed the prevalence of overt and occult hepatitis B virus (HBV) infection among leukemia patients. Among 256 leukemia patients and 377 fracture patients (control group), we found that the hepatitis B surface-antigen-positive rate was greater in leukemia patients than in the controls (odds ratio, 2.08; p = 0.01). Moreover, the prevalence of occult HBV infection was higher in leukemia patients than in the controls (10.5 % vs. 2.9 %; odds ratio, 3.92; p < 0.001). The HBV genotype distribution differed significantly between the leukemia and chronic hepatitis B or control groups (p < 0.001 and 0.01, respectively); specifically, genotype C was primarily observed in occult HBV infection patients with leukemia. The stop codon mutation rate or amino acid substitutions in the major hydrophilic region did not differ between the groups. Thus, the prevalence of occult hepatitis B is higher in leukemia patients, and the HBV genotype distribution differs between patients with leukemia and chronic hepatitis B virus infection.

In drug discovery and development, it is very important to investigate the plasma protein binding (PPB) of a drug to better understand its in vivo fate. In this study, a rapid and low-cost solid-phase extraction (SPE) method was developed for determining the PPB. With this method, the total protein recovery of a blank human plasma sample was 83.7 %. The unbound drug was easily adsorbed by an ODS C18 SPE column, and the recovery of three known drugs was more than 90 %. Their PPBs obtained by the SPE were identical to the value reported by conventional techniques. In addition, more than 90 % of 4-amino-2-trifluoromethyl-phenyl retinate (ATPR), which is a novel all-trans retinoic acid derivative (ATRA), was bound to human plasma protein as determined by SPE, and this value was comparable with that obtained by our previously described gel filtration-based method. Considering its versatility, speed of separation, and low cost, SPE is a rapid and economical method for measuring PPB.

•Novel dihydropyrazole as potential telomerase inhibitors were designed.•Compound 4f showed the most potent inhibitory activity.•Compounds could induce tumor cell apoptosis.A series novel 1-(3-substituted-5-phenyl-4,5-dihydropyrazol-1-yl)-2-thio-ethanone derivatives as potential telomerase inhibitors were designed and synthesized. The bioassays demonstrated that compounds 4a, 4f, 4j and 7b, 7d occupied high antiproliferative activity against SGC-7901, MGC-803, Bcap-37 and HEPG-2 cell lines. By a modified TRAP assay, some title compounds were tested against telomerase, and compound 4f showed the most potent inhibitory activity with IC50 value at 0.92 ± 0.09 μM. The mechanism of antitumor action indicated that title compounds 4f and 7b could suppress cell proliferation through inducing cell cycle arrest in G0/G1 phase.Novel 4,5-dihydropyrazol-1-yl-2-thio-ethanones as potential telomerase inhibitors were synthesized. The mechanism of the antitumor action indicated that title compounds could inhibit the growth of MGC-803 cells via the induction of tumor cell apoptosis.

To explore the characteristic expression of endoplasmic reticulum (ER) stress protein in antigen-induced arthritis models and the role of ER stress in arthritis.Effective animal models of rheumatoid arthritis in rabbits and rats were induced by methylated bovine serum albumin and Freund’s complete adjuvant. Pathological changes were assessed by magnetic resonance imaging and histological analysis. The expression and localization of ER stress proteins in synovium and peritoneal macrophages (PMΦ) were analyzed by double immunofluorescence staining. RT-PCR was performed to detect mRNA expression of ER stress-related genes. Tumor necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β) levels in synoviocytes were measured by RT-PCR and radioimmunoassay.We found that the ER stress marker BiP was highly up-regulated in arthritis synovium and extensively expressed in fibroblast-like synoviocytes (FLS) and macrophage-like synoviocytes (MLS). The expression of the pro-apoptotic factor CHOP/GADD153 was slightly elevated in inflammatory synovium and mainly localized in FLS, but insignificant in MLS. Unexpectedly, increased expression of CHOP was observed in PMΦ in arthritis rats. Likewise, cleaved caspase-3 was rarely expressed in MLS. In addition, induction of ER stress by tunicamycin resulted in significantly increased expression of pro-inflammatory molecules such as IL-1β and TNF-α in cultured inflammatory FLS.Differential activation of the ER stress proteins in synovium MLS may contribute to the resistance of synoviocytes to ER stress-induced apoptosis. Furthermore, ER stress is a potential mediator of arthritis inflammation.

A series of novel N-phenylacetyl (sulfonyl) 4,5-dihydropyrazole derivatives as potential telomerase inhibitors were synthesized. The bioassay tests show that compound 4a exhibited high activity against human gastric cancer cell SGC-7901, liver cancer Hep-G2 and human prostate PC-3 cell lines with IC50 values of 21.23 ± 0.99, 29.43 ± 0.32 and 30.89 ± 1.07 μM, respectively. All title compounds were assayed for telomerase inhibition by a modified TRAP assay, the results show that compound 4a can inhibit telomerase with IC50 value of 4.0 ± 0.32 μM. Docking simulation was performed to position compound 4a into the telomerase (3DU6) active site to determine the probable binding model.Novel 5-(2-hydroxyphenyl)-3-methyl-4,5-dihydropyrazol-1-yl-anone as potential telomerase inhibitors were synthesized. The bioassay tests show that compound 4a exhibited high activity against human gastric cancer cell SGC-7901, Hep-G2 and PC-3 cell lines. All title compounds were assayed for telomerase inhibition, the results show that compound 4a can inhibit telomerase. Docking simulation was performed to position compound 4a into the telomerase (3DU6) active site to determine the probable binding model.

The study was to evaluate the effect of triterpene acids of Eriobotrya japonica (thunb.) lindl. leaf (TAL) on inflammatory cytokine and mediator expression in alveolar macrophages (AM) of chronic bronchitic (CB) rats.CB was induced by endotracheal instillation of lipopolysaccharide (LPS) followed by Bacillus Calmette Guerin (BCG) injection via the caudal vein one week later. Treatment groups received TAL at there different doses (50, 150, or 450 mg/kg daily i. g.), Ketotifen fumarate (5 mg/kg daily i. g.) or dexamethasone (1.2 mg/kg daily i. g.) for two weeks, 7 days after LPS injection. AM were then isolated and incubated for 24 h. IL-1, TNF-α and PGE2 levels in cultured supernatants were measured by thymocyte co-stimulating assay and radioimmunoassay. Immunocytochemistry staining and western-blot were used for intracellular location and activation of p65 subunit of NF-kB. LTB4 level was analyzed by reverse-phase high performance liquid chromatography (RP-HPLC).The levels of TNF-α, IL-1, NF-kB, PGE2 and LTB4 expression in AM of TAL groups were significantly decreased compared to the CB group (P < 0.05 or P < 0.01), in a dose dependent manner.TAL inhibited NF-kB activation in AM from CB rats and led to down regulation of TNF-α, IL-1, PGE2 and LTB4 expression, which might be a mechanism for its anti-inflammatory effects in CB rats.

The aim of this study was to evaluate the association between macrophage migration inhibitory factor (MIF) −173G/C (rs755622), mannose-binding lectin (MBL2) exon 1 codon 54 (rs1800450) gene polymorphisms and rheumatoid arthritis (RA) susceptibility in ethnically different populations. A meta-analysis was conducted (allelic contrast, the additive model, the dominant model and the recessive model) on the MIF−173G/C polymorphism across five studies (four European and one Asian studies), and the MBL2 codon 54 polymorphism with five studies (four Asian and one European studies), respectively. Meta-analysis indicated an association between the MIF−173G/C in all study subjects in allelic contrast (OR = 1.19, 95%CI: 1.05–1.35, P = 0.001), the additive model (OR = 1.68, 95CI: 1.13–2.49, P = 0.001), the dominant model (OR = 1.17, 95CI: 1.01–1.35, P = 0.003), the recessive model (OR = 1.63, 95CI: 1.10–2.42, P = 0.001). While stratified by ethnicity with European populations, an association was found in allelic contrast (OR = 1.20, 95CI: 1.04–1.38, P = 0.002), the additive model (OR = 1.85, 95CI: 1.19–2.88, P = 0.001), the dominant model (OR = 1.20, 95CI: 1.02–1.41, P = 0.003). With respect to MBL2 codon 54 polymorphism and RA, no association was found in all study subjects in all comparisons, but there was an association while stratified by ethnicity with Asian populations in the dominant model (OR = 1.50, 95CI: 1.01–2.23, P = 0.007). In conclusion, the present study suggests that the MIF−173G/C polymorphism is associated with RA susceptibility, but the MBL2 codon 54 polymorphism is not associated with RA.

Activation of hepatic stellate cells (HSC) plays a pivotal role in the development of hepatic fibrosis. Transforming growth factor-β1 (TGF-β1) is considered to be the main stimuli factor responsible for the activation of HSC. MicroRNAs (miRNAs) have recently been shown to regulate cell proliferation, differentiation, and apoptosis. The involvement of miRNAs and their roles in TGF-β1-induced HSC activation remains largely unknown. Our study found that the expression of miR-146a was downregulated in HSC in response to TGF-β1 stimulation in dose-dependent manner by one-step real-time quantitative PCR. Moreover, we sought to examine whether miR-146a became dysregulated in CCl4-induced hepatic fibrosis in rats. Our study revealed that miR-146a was downregulated in liver fibrotic tissues. In addition, The HSC transfected with miR-146a mimics exhibited attendated TGF-β1-induced α-smooth muscle actin (α-SMA) expression compared with the control. Furthermore, overexpression of miR-146a suppressed TGF-β-induced HSC proliferation, and increased HSC apoptosis. Bioinformatics analyses predict that SMAD4 is the potential target of miR-146a. MiR-146a overexpression in TGF-β1-treated HSC did not decrease target mRNA levels, but significantly reduced target protein expression. These results suggested that miR-146a may function as a novel regulator to modulate HSC activation during TGF-β1 induction by targeting SMAD4.Highlights► The expression of miR-146a in HSC ► miR-146a modulates SMAD4 in HSC. ► miR-146a modulates HSC proliferation by targeting SMAD4.

ObjectiveTo study the effect of Wuziyanzong treatment on the sperm quality and content of calcium ions (Ca2+) in oligoasthenospermia rats.MethodsA model of oligoasthenospermia was induced in 50 Sprague Dawley rats by treatment with tripterygium glycosides at 30 mg/kg per day for 8 weeks. They were divided randomly into a model group, a positive group (Huangjingzanyu capsule, 3.01 g/kg), and low, medium and high dose Wuziyanzong treatment groups (2.30, 4.60, 9.20 g/kg crude drug respectively) with 10 in each group. Another 10 rats were used as a control group. The rats in the control and model groups were administered distilled water, while the rats in the remaining groups were administered Wuziyanzong for 30 d. The epididymides were removed, spermatozoa recovered and the sperm density and viability were measured. The spermatozoa were purified and the contents of Ca2+ in the cytoplasm and mitochondria were detected by flow cytometry and atomic absorption spectrometry, respectively.ResultsAfter 8 weeks of treatment with tripterygium glycosides, the sperm density, sperm activity and the Ca2+ content of spermatozoa in the model rats were all significantly decreased compared with the control group (all P<0.05). After 30 d treatment, the sperm density and activity improved and the Ca2+ content of sperm were increased significantly in the medium and high dose Wuziyanzong treatment groups in comparison with the model group (all P<0.05).ConclusionsThe Wuziyanzong treatment increased sperm density, improved sperm viability and enhanced the content of Ca2+ in the sperm cytoplasm and mitochondria in this rat model of oligoasthenospermia.

In this manuscript, we demonstrated that following stimulus by Kupffer cell-conditioned medium (KCM) and PDGF-BB, hepatic stellate cells (HSCs) showed significant increases in DNA synthesis and PDGFR-β expression. Furthermore, phosphorylation of PDGFR-β and three major members of the mitogen-activated protein kinase (MAPK) family were also significantly increased. Studies with respective neutralizing antibodies against released cytokines in conditioned medium demonstrated that PDGF-BB played an essential role in this complex activation process. Administration of A771726, leflunomide’s metabolite, markedly blunted these effects. However, the combination of A771726 with any of the three MAPK inhibitors potentiated this inhibitory effect and showed completely blockage on PDGFR-β expression and phosphorylation. Collectively, these data demonstrate that leflunomide inhibits KCM-mediated HSC proliferation via PDGFR-β phosphorylation and the subsequent activation of the MAPK pathway. Accordingly, targeted intervention against the PDGF-BB isoform may also offer a promising therapeutic approach to liver fibrosis.

AimTo investigate the therapeutic effects and mechanisms of total flavonoids of Turpinia Arguta Seen (TFS) on adjuvant arthritis in rats.MethodsThe model of adjuvant arthritis was induced by injection of Freund's Complete Adjuvant (FCA). Secondary paw swelling of AA rats was measured with volume meter and polyarthritis index were scored. The splenocyte proliferation, (interleukin-1) IL-1 and interleukin-2 (IL-2) production were assayed by cell proliferation assay. Prostaglandin E2 (PGE2) production was determined by radioimmunoassay.ResultsTFS (80, 160, 320 mg/kg, i.g.) could significantly inhibit secondary inflammatory reaction (secondary swelling, multiple arthritis, pathologic change of ankle arthritis) in AA rats. The results in vivo showed that the low response of splenocytes to concanavalin A (Con A) and lipopolysaccharide (LPS) and the decreased IL-2 synthesis were restored in AA rats treated with TFS (160, 320 mg/kg, i.g.), while the elevated IL-1 and PGE2 released from peritoneal macrophages (PMφ) were also reduced.ConclusionTFS has significant therapeutic effect on AA rats, which might be relate to its immunoregulatory actions.

Yupingfeng is a Chinese herbal compound used efficaciously to treat respiratory tract diseases. Total glucosides of Yupingfeng have been proven effective in anti-inflammation and immunoregulation. Nevertheless, the role of total extract of Yupingfeng (YTE) in pulmonary fibrosis (PF), a severe lung disease with no substantial therapies, remains unknown. Present study was conducted to elucidate the anti-fibrotic activity of YTE. The rat PF model was induced by intratracheal administration of bleomycin (BLM, 5 mg/kg), and YTE (12 mg/kg/d) was gavaged from the second day. At 14 and 28 days, the lungs were harvested and stained with H&E and Masson's trichrome. The content of hydroxyproline (HYP) and type I collagen (Col-I) were detected, while the protein expression of high-mobility group box 1 (HMGB1), transforming growth factor-beta 1 (TGF-β1), Col-I and α-smooth muscle actin (α-SMA) were analyzed by immunohistochemistry or Western blot. As observed, YTE treatment attenuated the alveolitis and fibrosis induced by BLM, reduced the loss of body weight and increase of lung coefficient. Meanwhile, YTE strongly decreased the levels of HYP and Col-I, and reduced the over-expression of HMGB1, TGF-β1, Col-I and α-SMA. In conclusion, YTE could ameliorate BLM-induced lung fibrosis by alleviating HMGB1 activity and TGF-β1 activation, suggesting therapeutic potential for PF.Download high-res image (179KB)Download full-size image