Simultaneous imaging of cancer biomarkers in live cells based on DNA-engineered exosomes.

Cancer biomarkers are directly related to the development of cancers. Noninvasive identification of the location and expression levels of these biomarkers in live cancer cells offers great potential for accurate early-stage cancer diagnosis and cancer metastasis monitoring. Herein, we propose a DNA-engineered exosome (DNA-Exo) nanoplatform to image dual cancer biomarkers at the single-cell level, in which DNA probes were modified with the cholesterol group to facilely anchor on the exosomal membrane through hydrophobic interaction. Fluorophore-labeled DNA aptamer and hairpin probes targeting two kinds of cancer biomarkers of transmembrane glycoprotein mucin 1 (MUC1) and cytoplasmic microRNA-21 (miR-21), respectively, were employed for convenient dual-fluorescence imaging of cancer cells. The cellular uptake of DNA-Exos induced the specific recognition of MUC1 and miR-21, allowing the acquisition of the expression levels and spatial distributions of these two biomarkers in three tested cell lines. Our work demonstrated that the proposed DNA-Exos with designable functions have the capacity to visually discriminate different cell types based on the specific recognition of analytes.

[Therapeutic effect of cinnamaldehyde on ulcerative colitis in mice induced by dextran sulfate sodium with Candida albicans colonization and its effect on dectin-1/TLRs/NF-κB signaling pathway].

To observe the efficacy of cinnamaldehyde on dextran sulfate sodium(DSS)-induced ulcerative colitis(UC) with Can-dida albicans(Ca) colonization and its effect on dectin-1/TLRs/NF-κB signaling pathway in mice. C57 BL/6 mice were randomly divided into normal group, DSS group, DSS+Ca group, cinnamaldehyde group and mesalazine group. Mice in DSS+Ca group were given Ca(1×10~8 CFU per mouse) through intragastrical administration for 4 consecutive days and then distilled water with 3.0% DSS for 7 consecutive days. In cinnamaldehyde group and mesalazine group, in addition to the induction method of the DSS+Ca group, mice were given 75 mg·kg~(-1) cinnamaldehyde and 200 mg·kg~(-1) mesalazine accompanied with 3.0% DSS for 7 consecutive days, respectively. Mice in normal group and DSS group were correspondingly administered with distilled water. The general conditions of the mice were observed daily, the diseased activity index(DAI) score was calculated, and fungal loads of feces were detected by plate method. The mice were sacrificed on day 12, colon length was measured, colon mucosa damage index(CMDI) score was calculated, and histopathological analysis was carried out by HE staining. Anti-saccharomces cerevisiae antibody(ASCA) and ß-1,3-glucan in serum, and TNF-α, IL-1ß, IL-6, IL-8, IL-10 in serum and colon tissue were detected by ELISA. The contents of ß-1,3-glucan and macrophage infiltration in colon tissues were examined by immunofluorescence staining. The protein expressions of dectin-1, TLR2, TLR4 and NF-κB were detected by Western blot and immunohistochemistry staining. The results showed that cinnamaldehyde could significantly improve the general conditions of UC mice with Ca colonization, decrease DAI and histopathological scores, reduce intestinal mucosal congestion, erosion and colon shortening, decrease Ca load in mouse feces and tissues, down-regulate the contents of ASCA and ß-1,3-glucan in serum, reduce the contents of TNF-α, IL-1ß, IL-6, IL-8 and increase IL-10 in serum and colon tissues, inhibit macrophages infiltration and down-regulate the protein expression of dectin-1, TLR2, TLR4 and NF-κB in colon tissue. These results suggested that cinnamaldehyde had a therapeutic effect on UC mice with Ca colonization, which might be related to the inhibition of Ca proliferation, the regulation of dectin-1/TLRs/NF-κB signaling pathways and the coordination of the balance between pro-inflammatory and anti-inflammatory factors.

Quantitative analysis of biologically active ingredients of Five Seeds Combo by liquid chromatography-quadrupole time-of-flight mass spectrometry for quality control of commercial herbal product.

Five Seeds Combo (wu zi yan zong wan) is a traditional Chinese herbal formula composed of fructus Lycii, semen Cuscutae, fructus Rubi, semen Plantaginis, and fructus Schisandrae. This herbal prescription has been developed into herbal products by many pharmaceutical manufacturers for treating age-related symptoms. The present study aims to develop an analytical method for the quality control of this herbal drug. Nine active ingredients including schisantherin A, schisandrin B, schisandrin, schisandrin A, quercitrin, betaine, verbascoside, hyperoside, and kaempferol were selected as the targeted analytes for the analysis. By using liquid chromatogram/quadrupole time-of-flight mass spectrometry (MS), the nine chemical compounds were determined simultaneously from the chromatogram. The parameters for MS were optimized by orthogonal array testing and the best condition of the MS for the determination of the nine marker compounds was found to be 175, 75, and 700 V for fragmentor, skimmer, and voltage of capillary, respectively. The method validation showed that this analytical method had high precision and sensitivity (limit of quantitation was smaller than 10 ng/mL for most of the analytes). The method was found to be able to demonstrate the quality of Five Seeds Combo from different manufacturers.

Involvement of the cerebral monoamine neurotransmitters system in antidepressant-like effects of a chinese herbal decoction, baihe dihuang tang, in mice model.

Baihe Dihuang Tang (BDT) is a renowned Chinese herbal formula which is commonly used for treating patients with mental instability, absentmindedness, insomnia, deficient dysphoria, and other psychological diseases. These major symptoms closely associated with the depressive disorders. BDT was widely popular use for treating emotion-thought disorders for many years in China. In the present study, the antidepressant-like effect of BDT in mice was investigated by using the forced swim test (FST) and the tail suspension test (TST). The underlying mechanism was explored by determining the effect of BDT on the level of cerebral monoamine neurotransmitters. BDT (9 and 18 g/kg, p.o. for 14 days) administration significantly reduced the immobility time in both the FST and the TST without changing locomotion in the open field-test (OFT). Moreover, BDT treatment at the dose of 18 g/kg inhibited reserpine-induced ptosis. Meanwhile, BDT enhanced 5-HT and NA levels in mouse cerebrum as well as decreased the ratio of 5-HT compared to its metabolite, 5-HIAA, (turnover, 5-HIAA/5-HT) after TST. The results demonstrated that the antidepressant-like effect of BDT is mediated, at least partially, via the central monoaminergic neurotransmitter system.

Radiofrequency ablation combined with transarterial chemoembolization in treatment of hepatocellular carcinoma adjacent to the second hepatic hilus.

PURPOSE: To explore the efficacy and safety of using radiofrequency ablation (RFA) combined with transarterial chemoembolization (TACE) for treating hepatocellular carcinoma (HCC) adjacent to the second hepatic hilus. METHODS: Between February 2011 and June 2013, 17 patients with HCC underwent combination therapy of TACE and RFA under DSA and CT guidance at our institution. The 17 patients had a total of 23 hepatic tumors, 17 of which were adjacent to the second hepatic hilus. RESULTS: TACE combined with RFA was performed successfully in all 17 patients with no mortalities or major morbidities. During the 1-month follow-up, tumors of 15 patients (88.2%) were completely ablated after one therapy session and 2 patients had detectable tumor residue. During the follow-up time period (range 6-52 months), local tumor progression developed in 1 patient (1/17, 5.9%) and both local tumor progression and new tumors appeared in 1 patient (1/17, 5.9%). Also, new tumors developed in the untreated portions of the liver in 8 patients (8/17, 47.1%). No distant metastasis was found. Of the 17 patients, 6 (35.3%) died due to tumor progression (3/17, 17.6%), liver failure (2/17, 11.8%), or massive hemorrhage of the gastrointestinal tract (1/17, 5.9%). The overall survival rates were 94.1% (16/17), 82.4% (14/17), and 61.8% (11/17) at 12, 18, and 24 months, respectively, and the median survival time was 25 months (95% CI 18-27). CONCLUSION: Treatment using combination of TACE and RFA is an effective and safe therapeutic strategy for treating HCC with tumor(s) adjacent to the second hepatic hilus.

Airway stent placement for direct cricothyroid membrane puncture anesthesia with airway stricture or fistula.

Context: Tracheal stent implantation with bronchoscope has become one of the main methods for the treatment of airway stenosis. Bronchoscopy is usually performed under local or general anesthesia. Aims: The aim of this study is to report our experience with direct cricothyroid membrane puncture anesthesia for airway stent placement. Materials and Methods: The medical records of 48 consecutive patients who underwent direct cricothyroid membrane puncture anesthesia for tracheal or bronchial stent placement were reviewed. The puncture site was chosen as the interspace between the thyroid cartilage and cricoid cartilage. Through the cricothyroid membrane, lidocaine was injected into the trachea to suppress the cough reflex. The airway stent was placed fluoroscopically at 48 patients at the trachea or bronchus. The success rate and complications of cricothyroid membrane puncture anesthesia were assessed. Results: Anesthesia was successfully performed in all 48 patients. Six patients needed 2-4 mL supplementary lidocaine by injection from the catheter or stent delivery system. Technical success of the stent insertion was achieved in all 48 cases. Small asymptomatic subcutaneous hemorrhage occurred at the puncture site in two patients, which disappeared 1 or 2 days after stent placement. Conclusion: Direct cricothyroid membrane puncture anesthesia for airway metal stent insertion is safe and effective.

Involvement of 5-HT-BDNF signaling axis in mediating synergistic antidepressant-like effects after combined administration of two oligosaccharide esters.

Potential mechanisms of depression involving herbal medicines and their specific compounds include elevated 5-HT level and downstream BDNF pathway. To identify potentially new combined therapeutic strategies, 3,6'-disinapoylsucrose (DISS) and tenuifoliside A (TFSA) have been observed to show antidepressant-like effects and its related 5-HT-BDNF pathway. We have tried to investigate whether combined administration of DISS and TFSA exerted more effective in the treatment of depression, as assessed through tail suspension test (TST) and forced swimming test (FST). In addition, we also analyzed the expression of three important proteins, cyclic adenosine monophosphate (cAMP) response element binding (CREB), brain-derived neurotrophic factor (BDNF), and cAMP-regulated transcriptional coactivators (CRTC1), which have been shown to be involved in the regulation of the neurotrophic factors in the hippocampus. The DISS and TFSA separately, both at a dose of 5 mg/kg each, displayed small effect in the immobility time. However, combined treatment of these two in multiple doses exhibited better effect. Moreover, combined treatment of DISS and TFSA also demonstrated enhanced levels of 5-hydroxytryptamine (5-HT), and stronger increase in the phosphorylation levels of CREB, BDNF, and CRTC1 proteins in the hippocampus. Overall, our results indicated that coadministration of these two oligosaccharide esters at low dose may induce more pronounced antidepressant activity, in comparison with individual treatment even at high dosage. Thus, the antidepressant properties of both these compounds can be attributed to their ability to influence 5-HT and BDNF pathway, and thereby suggesting that this combination strategy can definitely act as alternative therapy for depression disorder with very limited side effects.

Incidence and risk of significantly raised blood pressure in cancer patients treated with bevacizumab: an updated meta-analysis.

PURPOSE: Bevacizumab is a humanized monoclonal antibody targeting vascular endothelial growth factor. The aim of this study was to gain a better understanding of the overall incidence and risk of significantly raised blood pressure in cancer patients who receive bevacizumab therapy. METHODS: We performed a meta-analysis of relevant randomized controlled trials (RCTs) identified in PubMed, Cochrane library, Embase, and American Society of Clinical Oncology conferences. Overall incidence rates, relative risks (RRs), and 95% confidence intervals (CIs) were calculated using a random-effects model. The primary clinical endpoint was significantly raised blood pressure (grade 3 or above). RESULTS: A total of 12,949 cancer patients with a variety of solid tumors from 19 RCTs were included in our meta-analysis. The overall incidence of significantly raised blood pressure was 8% (95% CI 6-10%) among patients receiving bevacizumab. Bevacizumab treatment was associated with a statistically significant increased risk of developing significantly raised blood pressure (RR 5.38, 95% CI 3.63-7.97). The RRs of significantly raised blood pressure in patients receiving bevacizumab at 5 and 2.5 mg/kg per week were 7.17 (95% CI, 3.91-13.13) and 4.11 (95% CI 2.49-6.78), respectively. Among cancer patients, those with renal cell carcinoma (RR 13.77, 95% CI 2.28-83.15) and breast cancer (RR 18.83, 95% CI 1.23-292.29) who received bevacizumab at 5 mg/kg per week had a higher risk of developing significantly raised blood pressure. CONCLUSIONS: Among the patients included in the trials analyzed in this meta-analysis, the addition of bevacizumab to cancer therapy treatments significantly increased the risk of significantly raised blood pressure. The risk may be dose-dependent and vary with tumor type.

Chemical and biological differentiation of Cortex Phellodendri Chinensis and Cortex Phellodendri Amurensis.

The Chinese herbal drug Cortex Phellodendri is derived from two species of PHELLODENDRON, P. CHINENSIS Schneid. and P. AMURENSE Rupr. Traditionally, Cortex Phellodendri Chinensis (CPC) and Cortex Phellodendri Amurensis (CPA) are used interchangeably because they are believed to share the same clinical efficacy. Berberine has been believed to be the active ingredient of the herbs. However, recent studies have shown that the content of berberine is much higher in CPC than in CPA. Interestingly, the majority of researches deal with CPA, the one with lower content of berberine. These observations provoke us to reconsider the active ingredients of Cortex Phellodendri. In this study, two traditional usages (antidiarrheal and antibacterial) of Cortex Phellodendri were compared with the chemical analysis of the two herb species used in its formulation. The results suggest that berberine is one of the active ingredients responsible for the antidiarrheal and antibacterial activities of the herbs, but that other chemical ingredients are also involved in regulating the biological actions of the herbal drug. These chemical ingredients may have the same or the opposite effect as berberine. The effectiveness of the herbs is more likely to correlate to the content of total alkaloids rather than to the content of berberine.

Ertapenem versus piperacillin/tazobactam for the treatment of complicated infections: a meta-analysis of randomized controlled trials.

BACKGROUND: Ertapenem, a new carbapenem with a favorable pharmacokinetic profile, has been approved for the treatment of complicated intra-abdominal Infections (cIAIs), acute pelvic infections (APIs) and complicated skin and skin-structure infections (cSSSIs). The aim of this study is to compare the efficacy and safety of ertapenem with piperacillin/tazobactam, which has been reported to possess good efficacy for the treatment of these complicated infections. METHODS: We performed a meta-analysis of randomized controlled trials identified in PubMed, Cochrane library and Embase that compared the efficacy and safety of ertapenem with piperacillin/tazobactam for the treatment of complicated infections including cIAIs, APIs, cSSSIs. The primary efficacy outcome was clinical treatment success assessed at the test-of-cure visit. The primary safety outcome was drug related clinical and laboratory adverse events occurred during the treatment and the post-treatment period. RESULT: Six RCTs, involving 3161 patients, were included in our meta-analysis. Ertapenem was associated similar clinical treatment success with piperacillin/tazobactam for complicated infections treatment (clinically evaluable population, 1937 patients, odds ratios: 1.15, 95% confidence intervals: 0.89-1.49; modified intention to treat population, 2855 patients, odds ratios: 1.03, 95% confidence intervals: 0.87-1.22). All of secondary efficacy outcomes analysis obtained similar findings with clinical treatment success. No difference was found about the incidence of drug related adverse events between ertapenem and piperacillin/tazobactam groups. CONCLUSION: This meta-analysis provides evidence that ertapenem 1 g once a day can be used as effectively and safely as recommended dose of piperacillin/tazobactam, for the treatment of complicated infections, particularly of mild to moderate severity. It is an appealing option for the treatment of these complicated infections.

Stimulating effect of catechin, an active component of Spatholobus suberectus Dunn, on bioactivity of hematopoietic growth factor.

BACKGROUND: Hematopoietic growth factor (HGF) is indispensable to hematopoiesis in the body. The proliferation and differentiation of hematopoietic cells must rely on the existence and stimulation of HGF. This study investigated the effect of catechin, an active component extracted from Spatholobus suberectus Dunn (SSD), on bioactivity of granulocyte-macrophage colony-stimulating activity (GM-CSA), burst-promoting activity (BPA) and megakaryocyte colony-stimulating activity (MK-CSA) in spleen condition medium (SPCM) of mice to clarify the hematopoietic mechanism of catechin and SSD. METHODS: Spleen cells of mice were separated and spleen condition medium (SPCM) was prepared from spleen cell culture. Bone marrow cells of mice were separated and cultured in a culture system including 10% (v/v) SPCM (induced by catechin in vivo or ex vivo) for 6 days. Granulocyte-macrophage colony forming units (CFU-GM), erythrocyte burst-colony-forming units (BFU-E) and megakaryocyte colony-forming units (CFU-Meg) formation were employed to assay the effects of different treatment on the bioactivity of GM-CSA, BPA and MK-CSA in SPCM. RESULTS: SPCM induced by 100 mg/L catechin ex vivo could promote the growth of CFU-GM, BFU-E and CFU-Meg, which indicated that catechin could stimulate the production of GM-CSA, BPA and MK-CSA in SPCM. SPCM prepared at the fourth day of spleen cell culture showed the best stimulating activity. The bioactivity of GM-CSA, BPA and MK-CSA in the SPCM prepared after intraperitoneally injecting catechin into mice was also increased. The number of CFU-GM, BFU-E and CFU-Meg gradually increased as the dose of catechin increased and the time of administration prolonged. CFU-GM, BFU-E and CFU-Meg of the high-dose catechin group were significantly higher than those of the control group (P < 0.01) and reached the maximum at the seventh day after administration. CONCLUSIONS: This study suggests that catechin extracted from the active acetic ether part of Spatholobus suberectus Dunn can regulate hematopoiesis by inducing bioactivity of GM-CSA, BPA and MK-CSA in SPCM of mice. This may be one of the mechanisms for the hematopoietic-supportive effect of catechin and Spatholobus suberectus Dunn.

[Effects of the Farmland-to-Forest/Grassland Conversion Program on the Soil Bacterial Community in the Loess Hilly Region].

This study investigated the effects of the program aimed at converting farmland into forest or grassland on the soil bacterial diversity in the Loess Hilly region. Corn land was selected as the experimental control, and Caragana intermedia land and Stipa bungeana land were selected as the experimental lands. Soil from three different land use types were selected as subjects. The soil bacterial communities were analyzed using a high throughput sequencing technique (MiSeq). The sequence region was 16S rRNA V3-V4 variable region. The α diversity, community composition, and relative abundances of the soil bacterial groups were analyzed, in order to explore the effects of soil physical and chemical properties on the bacterial community structure. The results showed that the structure and diversity of the microbial communities differed under the different land use types. At the phylum level, the dominant phyla were Proteobacteria, Actinobacteria, Acidobacteria, Chloroflexi, Gemmatimonadetes, and Firmicutes. At the class level, α-Proteobacteria, Actinobacteria, Acidobacteria, ß-Proteobacteria, Thermoleophilia, Gemmatimonadetes, Bacilli, and δ-Proteobacteria were predominant. Compared to the corn land, Proteobacteria and Actinobacteria were the dominant bacteria in the Caragana intermedia land, while Actinobacteria and Acidobacteria were the dominant bacteria in the Stipa bungeana land. Soil bacterial diversity was the highest in the shrub land. RDA analysis revealed that soil organic matters and total nitrogen were the most influential environmental factors. It was shown thus that the program of converting farmland into forest or grassland has significantly improved the soil fertility and environmental conditions, and the composition of the soil bacterial community has also been appreciably changed.

[Effect of catechin on bone marrow cell cycle and gene expression of hematopoietic growth factors].

AIM: To study the effect of catechin, the active component of Spatholobus suberectus Dunn, on bone marrow cell cycle and the expression of IL-6 and GM-CSF mRNA in spleen cells of normal and marrow-depressed mice in order to clarify the mechanism of hematopoietic-supportive effect of catechin. METHODS: Flow cytometry was adopted to investigate the influence of catechin on bone marrow cell cycle in mice and the expression of IL-6 and GM-CSF mRNA induced by catechin in spleen cells was detected by RT-PCR technique simultaneously. RESULTS: The cell proportion of normal and marrow-depressed mice in G0/G1 phase was reduced significantly, while that in S + G2/M phase increased significantly. Being induced by catechin, IL-6 mRNA and GM-CSF mRNA in spleen cells were markedly up-regulated. CONCLUSION: Catechin (2 g x L(-1), intraperitoneally injected to mice daily immediately after irradiation for 7 consecutive days) was shown to promote the expression of IL-6 mRNA and GM-CSF mRNA in spleen cells of mice, through which it can accelerate bone marrow cells of normal mice into cell cycle and help those of marrow-depressed mice to get out of "G1-phase-block", enter into cell cycle and radically accelerate the proliferation and differentiation of hematopoietic stem cell/hematopoietic progenitor cell (HSC/HPC).

[Effect of SS8, the active part of Spatholobus suberectus Dunn, on proliferation of hematopoietic progenitor cells in mice with bone marrow depression].

OBJECTIVE: To investigate the effect of SS8, a monomer extracted from Spatholobus suberectus Dunn used for invigorating the circulation of blood, on proliferation of hematopoietic progenitor cells in mice with bone marrow depression. METHOD: The method of semi-solid culture with methylcellulose of CFU-GM, CFU-E, BFU-E, CFU-Meg was adopted in bone marrow depressed mice which were treated with SS8 for a long time. RESULT: The experimental data demonstrated that the numbers of CFU-GM, CFU-E, BFU-E, CFU-Meg in bone marrow depressed mice were raised distinctly under the control of SS8 as compared with those of contrast group. The effect of SS8 became stronger as time went on and the dosage rose. CONCLUSION: SS8 can distinctly stimulate the proliferation of hematopoietec progenitor cells in mice with bone marrow depression in a time-and-dosagedependent manner.

[Effects of yishenjiangyafang on blood pressure and protecting renal function in RPH rats].

OBJECTIVE: To study the effects and mechanism of Yishenjiangyafang, a prescription of Chinese traditional herbs used for renal hypertension, on blood pressure and protecting renal function of RPH rats. METHOD: The 5/6 kidney of rat was resected to set up the RPH rat model. Blood pressure, Cr(creatinine), Ccr(creatinine clearance) and BUN(urea nitrogen) were measured dynamically. After eight weeks treatment, plasma content of PAR A II TXB2, 6-keto-PGF1 were measured. At same time, The change of renal pathology was observed. RESULT: Yishenjiangyafang could reduce blood pressure Cr, but had no effect on BUN of RPH rat. The indexes of PAR, A II of each different dosage group of Yishenjiangyafang were decreased. At the same time, it reduced plasma content of TXB2, and increased 6-keto-PGF1 alpha. Glomerulosclerosis and atrophy of renal tubule in Yishenjiangyafang group RPH rats were better than those of the contrast group and the Capten group. CONCLUSION: Yishenjiangyafang can reduce blood pressure of RPH rats and has protective effects on its kidney. Yishenjiangyafang can perform its effects of reducing blood pressure and protecting kidney by influencing the RAS of RPH rats.

Efficacy and safety of micafungin for invasive candida infections: a meta-analysis of randomized controlled trials.

BACKGROUND: Invasive fungal infections such as candidiasis and mold infections cause significant morbidity and mortality in seriously ill patients. Micafungin is an echinocandin antifungal agent with potent activity against most species of Candida and Aspergillus. We did this meta-analysis to clarify whether micafungin offers superior efficacy and safety compared with other antifungal agent for treating infections associated with invasive candidiasis. METHODS: We did a meta-analysis of randomized controlled trials to examine whether micafungin has superior efficacy and safety compared with other antifungal agents recommended by the treatment guidelines for fungal infection. Seven trials involving 2913 patients were included in this analysis. Odds ratios (OR) and 95% confidence intervals (CI) were calculated. RESULTS: Micafungin was associated with significantly better treatment success compared with the comparator antifungal agents (modified intention to treat, 2851 patients; random-effects model, OR 1.20, 95%CI 1.00 - 1.45, P = 0.0487). In addition, micafungin was more effective than the comparators for antifungal prophylaxis of neutropenic patients undergoing hematopoietic stem cell transplantation (OR 1.47, 95%CI 1.08 - 2.00, P = 0.01). Although there was no significant difference between the compared regimens in terms of the incidence of adverse drug effects (OR 0.94, 95%CI 0.77 - 1.11), fewer patients treated with micafungin withdrew from the studies because of adverse events (OR 0.64, 95%CI 0.44 - 0.94). CONCLUSIONS: Micafungin has a good safety and tolerability profile, with an efficacy at least comparable to the other antifungal agents. Micafungin offers advantages over other agents for antifungal prophylaxis. Micafungin offers an appropriate alternative for antifungal prophylaxis rather than the treatment of invasive candida infections.

Biochemical mechanism of Wu-Zi-Yan-Zong-Wan, a traditional Chinese herbal formula, against alcohol-induced oxidative damage in CYP2E1 cDNA-transfected HepG2 (E47) cells.

AIM OF THE STUDY: Wu-Zi-Yan-Zong-Wan (WZ) is a traditional Chinese herbal formula which is commonly used for treating patients with "Yang deficiency". In the present study, the effect of WZ on ethanol-induced toxicity in CYP2E1 cDNA-transfected HepG2 (E47) cells was investigated. MATERIALS AND METHODS: WZ extract was obtained by extracting the herbal powder with 50% ethanol (v/v, in water) and the effect of the extract on ethanol-induced toxicity was investigated in cultured cells. RESULTS: The treatment with WZ extract (12.5-200 microg/mL) for 24h dose-dependently protected against ethanol-induced toxicity in E47 cells, as evidenced by the enhanced cell viability and decreased extent of lactate dehydrogeanse leakage. The cytoprotection against ethanol-induced toxicity was associated with decreases in the extents of reactive oxygen species production and lipid peroxidation, as well as increases in mitochondrial reduced glutathione and membrane potential. In addition, WZ extract treatment also suppressed the formation of DNA fragments in ethanol-intoxicated E47 cells. CONCLUSIONS: WZ extract was found to protect against the ethanol-induced toxicity in E47 cells, possibly by virtues of its antioxidant activity.

Long-term treatment with a "Yang-invigorating" Chinese herbal formula, Wu-Zi-Yan-Zong-Wan, reduces mortality and liver oxidative damage in chronic alcohol-intoxicated rats.

Long-term alcohol consumption has been reported to increase oxidative stress in multiple organs and accelerate the aging process. A previous study in our laboratory has shown that Wu-Zi-Yan-Zong-Wan (WZ), a "Yang-invigorating" Chinese herbal formula, protected against ethanol-induced toxicity in HepG2 cells transfected to express human CYP2E1, presumably by enhancing mitochondrial antioxidant status and functional ability. The present study aims to investigate whether WZ extract treatment can afford protection against chronic ethanol-induced oxidative stress (a major risk factor of aging) and mortality in rats. The effect of the extract (1.8 g, 4.5 g, and 9 g raw materials/kg per day) on chronic ethanol hepatotoxicity was investigated in rats receiving steady intragastric infusion of ethanol-containing liquid diet. The results showed that long-term (42 days) herbal co-treatment protected against chronic ethanol-induced mortality and hepatotoxicity and in rats, as evidenced by decreased plasma transaminases activities. The extract also suppressed the pathological development of fatty liver, as assessed by histopathological examination and the ratio of liver weight to body weight. The hepatoprotection afforded by the extract was associated with decreases in the extents of reactive oxygen species production, lipid peroxidation, and oxidative modification of proteins, as well as the reversal of altered mitochondrial reduced glutathione level. The results suggest that the suppressive effect of WZ on chronic ethanol-induced oxidative stress and mortality may be attributed to the antioxidant action, particularly in mitochondria.