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The trypsin-assisted extraction of polysaccharides from Allium cepa L. was optimized using the response surface methodology (RSM). The optimum extraction conditions were extraction temperature, ext...

Undaria pinnatifida (U. pinnatifida) polysaccharides (UPPS) are considered to be the major bioactive components of U. pinnatifida. The aim of the present study was to investigate the separation, sulfated modification, characterization and monosaccharide composition of UPPS. The optimal processing conditions were as follows: Distilled water-to-solid ratio, 50 ml/g; extraction time, 300 min; and extraction temperature, 90˚C. The major polysaccharide fraction of U. pinnatifida (UPPS-B1) was purified via DEAE-52 and Sephadex G-200 column chromatography. The chlorosulfonic acid-pyridine method was applied for sulfation modification. UPPS-B1 and sulfated (S)-UPPS-B1 were characterized via chemical analysis, ultraviolet-visible and Fourier-transformed infrared spectroscopy, gas chromatography and high-performance liquid chromatography. The total sugar content of UPPS-B1 and S-UPPS-B1 was 79.78 and 77.28%, respectively. The sulfate radical content of UPPS-B1 and S-UPPS-B1 was 8.53 and 29.12%, whilst the content of uronic acid was 9.29 and 7.98%, respectively. The average molecular weight of UPPS-B1 and S-UPPS-B1 was determined to be 37 and 110 kD, respectively. UPPS-B1 was considered to be a heteropolysaccharide composed of xylose, mannose, glucose and galactose at a ratio of 7.9:8.7:12.0:9.8. In addition, S-UPPS-B1 was a heteropolysaccharide composed of xylose, mannose, glucose and galactose at a ratio of 1.0:9.7:6.4:1.6. The results of the tumor growth inhibition experiment demonstrated that UPPS-B1 exhibited anti-tumor activity in vivo, which was improved following sulfation to yield S-UPPS-B1.

The main objective of this study is to establish the best preparation technology of the particles of Acanthopanax senticosus. First, take the reflux extraction method extract of Acanthopanax senticosus coarse powder, optimized by orthogonal experimental method, to flavonoids flavonoids extraction extraction rate as the indexes to determine the effects of extraction temperature, ethanol concentration, extraction time on flavonoids content. Then by a wet granulation of thorn slender acanthopanax particles, taste with granules, forming rate, melting rate as index to investigate the influences of materials adding amount of granules effect. The results showed that the ethanol water heating reflux extraction method to extract the temperature of 70 deg, the percentage of ethanol 75%, extraction time 2.5 h, the highest content of total flavonoids in the extract. Join the 5 ml and 10 g in the extract of acacia honey, dextrin, starch, sugar ratio for 3:4:8, the best taste of Acanthopanax granules. In the end, the best preparation technology of the granules is established, and the process is simple, which is suitable for the large-scale production of the factory. Keywords: Acanthopanax senticosus, Total flavonoids, Extraction technology, Preparation technology

Gynura procumbens is commonly consumed as a vegetable and has been approved as an ingredient for food and dietary supplements in China. However, Gynura species are known to contain toxic py...

LC technology is a recognized method used worldwide to evaluate the quality of traditional Chinese medicines (TCM). The quality of TCM has a direct impact on its efficacy. Therefore, in order to thoroughly reveal how TCM exerts its efficacy, first of all, it is necessary to understand the material basis for its efficacy, and then to control the quality of active compounds. The application of the spectrum-effect relationship method is crucial for determining the pharmacological material basis. The goal of this paper was to investigate the underlying correlations between the chemical profiles and oestrogenic activity of Cistanche, to reveal the active compounds. The chemical profiles of Cistanche were recorded using HPLC/Q-TOF-MS/MS, and oestrogenic activity was determined by the Uterus growth test and the MTT assay. Then combining the results of bivariate analysis, principal component analysis and gray correlation analysis method, fifteen active compounds were identified. They are 8-epiloganic acid, salidroside, syringalide A 3’-α-l-rhamnopyranoside, cistanoside A, echinacoside, cistanoside F, cistanoside B, cistanoside C, osmanthuside B, acteoside, isoacteoside, tubuloside B, 2’-acetylacteoside, and two unknown compounds. This study lays a foundation for in vivo studies of Cistanche and for the development of its clinical application.

The trypsin-assisted extraction of polysaccharides from Allium cepa L. was optimized using the response surface methodology (RSM). The optimum extraction conditions were extraction temperature, extraction time, extraction pH, and enzyme amount of 37.16°C, 180 min, 8.57, and 5.16%, respectively. Under the optimized conditions, the yield of A. cepa L. polysaccharides (ACP) reached 9.69%, which was comparable with the predicted yield (9.73%). Mid- and high-dose ACP significantly inhibited the tumor growth (43.93%) and the tumor inhibition percentage (38.05%), which were more than 30%. The ACP could extend the survival time of H22 ascites tumor-bearing mice. Furthermore, the ACP could reduce the thymus and the spleen atrophy and significantly promoted the Con A-induced proliferation of splenocytes and elevated the serum IFN-γ and IL-2 levels. Therefore, the ACP could inhibit the tumor growth in tumor-bearing mice and regulated the immune function of mice. Practical ApplicationsThe trypsin-assisted extraction has high efficiency, is carried out through the polysaccharide extraction and the deproteinization at the same time, and is more convenient and fast than traditional methods. No detailed study on the optimization of the trypsin extraction of onion polysaccharides is available. Thus, this experiment aims to use the BBD (4 factors and 3 levels) to optimize the roles of extraction temperature, extraction time, extraction pH, and amount of enzyme on the yield of polysaccharides obtained from the fruit of A. cepa L. In addition, when looking for high-quality biological functional principles for the pharmaceutical industry, the antitumor activity of ACP was evaluated. A. cepa L. is one of the most widely cultivated and consumed crops worldwide. Polysaccharides are the main active ingredient, and studies have shown that a high intake of Allium vegetables is associated with reduced risk of cancers.

Strain SZW01 was isolated from sea sediment collected from Shenzhen in Guangdong province, China, and was later identified as

The extract of the strain Aspergillus flavipes DL-11 exerted antibacterial activities against six Gram-positive bacteria. During the following bioassay-guided separation, ten diphenyl ethers (1-10), two benzophenones (11-12), together with two xanthones (13-14) were isolated. Among them, 4'-chloroasterric acid (1) was a new chlorinated diphenyl ether. Their structures were elucidated by extensive spectroscopic data analysis, including IR, HR-ESI-MS, NMR experiments, and by comparison with the literature data. All compounds showed moderate to strong antibacterial effects on different Gram-positive bacteria with MIC values that ranged from 3.13 to 50 μg/mL, but none of the compounds exhibited activity against Gram-negative bacteria Vibrio parahaemolyticus ATCC17802 (MIC>100 μg/mL). In particular, the MICs of some compounds are at the level of positive control.

Neutrophils are key inflammatory cells in the immunopathogenesis of asthma. Neutrophil migration can be initiated through activation of the CXCR1 and CXCR2 receptors by CXC chemokines, such as IL-8. Although transcription factor KLF2 has been found to maintain T cell migration patterns through repression of several chemokine receptors, whether KLF2 can regulate neutrophil migration via modulation of CXCR1 and CXCR2 is unknown. Here, we aimed to explore the functions of KLF2, CXCR1 and CXCR2 in neutrophil migration in asthma and to establish a regulatory role of KLF2 for CXCR1/2. We demonstrate that with asthma aggravation, the percentages and migration rates of peripheral blood neutrophils gradually increased in asthmatic patients and the guinea pig asthma model. Correspondingly, both the KLF2 mRNA and protein levels in neutrophils were gradually reduced. While CXCR1 and CXCR2 expression was negatively correlated with KLF2. In vitro knockdown of KLF2 dramatically increased the migration of HL-60-drived neutrophil-like cells, which was accompanied by an increase in the CXCR1 and CXCR2 mRNA and protein expression levels. Taken together, our results indicate that decreased KLF2 aggravates asthma progression by promoting neutrophil migration, which is associated with the transcriptional upregulation of CXCR1 and CXCR2. The KLF2 and/or CXCR1/2 expression levels may represent an indicator of asthma severity.

Cytisine is a quinolizidine alkaloid, which has been reported to be among the major bioactive components of Sophora alopecuraides L. Quinolizidine alkaloids have previously been demonstrated to inhibit the proliferation of several types of tumor cells. However, few studies have investigated the effects of cytisine on cancer cells. The present study was performed to further investigate the molecular mechanisms underlying cytisine‑induced apoptosis of HepG2 human hepatocellular carcinoma cells. The results of an MTT assay demonstrated that cytisine inhibited the growth of HepG2 cells in a dose‑dependent manner. In addition, the induction of apoptosis was detected, as determined by morphological observation and flow cytometry. As determined by fluorescence microscopy, apoptotic morphological alterations were detected following cytisine administration. Flow cytometric analyses demonstrated that cytisine induced cytotoxicity through apoptosis‑like mechanisms in HepG2 cells. Furthermore, western blot analysis was performed to investigate the release of cytochrome c (Cyt‑c) and activation of the caspase cascade, and the results indicated that treatment of HepG2 cells with cytisine induced caspase‑dependent apoptosis via the release of Cyt‑c from the mitochondria, upregulation of caspase‑3 and downregulation of pro‑caspase‑3. These results indicated that cytisine may induce apoptosis of HepG2 cells through the mitochondrial pathway.

In this experiment,the antioxidant capacity of raspberry extract and the protective effect on liver injury induced by ConA in mice were investigated. Balb/C male mice were randomly divided into six groups: normal group,model group,bicyclol control group( 200 mg·kg~(-1)),low-dose raspberry extract group( 200 mg·kg~(-1)),middle-dose raspberry extract group( 400 mg·kg~(-1)),and highdose raspberry extract group( 800 mg·kg~(-1)). Each group was intragastrically administered with drugs according to the body weight once a day. Seven days later,all of the groups except for the normal group were treated with ConA( 20 mg·kg~(-1)) through tail vein injection to establish the acute liver injury model. The mice were put to death 8 hours later. The organ indexes were calculated. These rum levels of ALT,AST and LDH and the activities of SOD,CAT,GSH and MDA in liver tissue were detected. HE staining was used to observe the pathological changes of liver tissue in mice. Western blot was used to detect the expressions of Bax,Bcl-2,Nrf2 and Keap-1. The antioxidant capacity of raspberry extract was measured by CAA assay. The results showed that,raspberry extract had a strong antioxidant capacity. Simultaneously,compared with the model group,raspberry extract can significantly improve the pathological conditions of liver,and significantly reduce ALT,AST and LDH activities in serum of liver injury mice( P0. 01). The activities of SOD,CAT in liver homogenate supernatant were significantly increased in the high-dose group,the content of GSH increased,while the content of MDA was sharply declined in the high-dose group( P0. 01). Meanwhile,raspberry extract down-regulated the expressions of Bax and Keap-1 and up-regulated the expressions of Bcl-2 and Nrf2. CAA showed that the compound raspberry extract had a strong antioxidant capacity. Therefore,raspberry extract has an obvious protective effect on acute liver injury induced by ConA in mice.

Evodia rutaecarpa (E. rutaecarpa) has been used to treat aches, vomiting and dysentery in traditional Chinese medicine. However, as a mildly toxic herb its toxic components have not been elucidated. An attempt was made to illuminate the hepatotoxic constituents of E. rutaecarpa. The 50% ethanol extracts of E. rutaecarpa from 19 different sources were used to establish UPLC fingerprints and administered to mice at a dose of 35 g/kg (crude medicine weight/mouse weight) once daily for 14 days. Serum levels of alanine transaminase, aspartate aminotransferase and liver coefficient were used as indices of liver injury. Additionally, the characteristic peaks of 19 fingerprints were identified. Spectrum-effect relationships between fingerprints and hepatotoxic indicators were analyzed using bivariate correlation analysis (BCA). The UPLC fingerprints were established and a total of 28 main compounds were identified. Because of the inherent variations in chemical compositions, the liver injury levels were different among the E. rutaecarpa samples from 19 sites of production. BCA results indicated that compounds dihydrorutaecarpine, 6-acetoxy-5-epilimonin, goshuyuamide I, 1-methyl-2-[(Z)-5-undecenyl]-4(1H)-quinolone, 1-methyl-2-[(4Z,7Z)-4,7-tridecadienyl]-4(1H)-quinolone, evocarpine and 1-methyl-2-[(6Z,9Z)-6,9-pentadecadienyl]-4(1H)-quinolone were tentatively determined as the primary hepatotoxic components. The present study provides a valuable method for the discovery of hepatotoxic constituents by combination of fingerprints and hepatotoxicity index.

Cajaninstilbene acid (CSA) exerts wide pharmacological activities, such as anti-inflammation, hypoglycaemic activity, analgesic effect and cognition improvement. However, it underwent severe phase II metabolism mediated by UDP-glucuronosyltransferase (UGT) in the gastrointestinal (GI) tract after oral administration, affecting its oral bioavailability. In the present study, we utilize UGT inhibitory excipient containing self-microemulsion (SME) delivery system to reduce the production of glucuronide metabolites and increase its oral bioavailability. The present results showed that although similar properties in physiochemical, cytotoxicity, cellular uptake, absorption and transport across rat everted gut sacs between SME-1 (inhibitory excipient containing SME) and SME-2 (control SME, without inhibitory excipient), an improved absolute bioavailability of 57.3 % was conferred by SME-1, significantly higher than the value of 35.4 % by SME-2 and 34.0 % by free CSA. Noticeably, the significantly lower AUC value of CSA glucuronide was determined in rats treated with SME-1 than those either treated with SME-2 or free CSA. Thus, the ability of SME-1 to enhance oral bioavailability of CSA is mainly attributed to the inhibition of phase II metabolism in the GI tract.

High-performance liquid chromatography (HPLC) is an efficient method that is widely used to assess the quality of traditional Chinese medicine (TCM). It is well known that the quality of TCM has a direct effect on its efficacy; therefore, in order to thoroughly explain how TCM exerts its efficacy, it is necessary to characterize its active ingredients and assess their quality. The application of the spectrum‑effect method is crucial for determining the pharmacological basis of materials. The aim of the present study was to examine the correlation between chemical spectra and estrogenic activity of Cuscuta chinensis Lam., in order to reveal active compounds with potential therapeutic effects. The spectra of Cuscuta chinensis Lam. were recorded using HPLC, and estrogenic activity was determined using a uterus growth test and MTT assay. Combination of the results of bivariate analysis, principal component analysis and Gray relational analysis identified 19 active compounds, as follows: Quercetin‑3‑O‑(2'‑O‑α‑rhamnosy‑6'‑O‑malony)‑​β‑D‑glucoside, ka-empferol‑3‑O‑β‑D‑aplosyl‑(1→2)‑​[‑α‑​L‑rhamnosy‑(1→6)]‑β-wD-glucoside, 6‑O‑(E)‑P‑coumaroyl)‑β‑​D‑fructofuranosyl‑(2→1)‑α‑D‑glucopyranoside, kaempferol‑7‑rhamnosy, kaempferol‑3‑β‑D-glucuronide, apigenin, 4‑caffeoyl‑5‑coumaroylquinic acid, kaempferol‑3‑arabofuranoside, quercetin‑3‑O-β‑D-apiofuranosyl-(1→2)-β‑D‑galactoside, dicaffeoylquinic acid, hyperin, quercitin, isorhamnetin, chlorogenic acid, quercetin, quercltrin‑2''‑gallate, quercetin‑3, 7‑α‑L‑dirhamnoside and stigmasterol, as well as one unknown compound. The present study laid a foundation for in vivo metabolic studies regarding Cuscuta chinensis Lam. and for the development of its clinical application.

The caper plant (Capparis spinosa L.) was a common Uyghur folk medicine, and is a member of the Capparidaceae family. In a previous study, the n-butanol extract of C. spinosa L. (CSBE) was demonstrated to exert anti-tumor activity; however, the underlying mechanism is currently not understood. The present study aimed to elucidate the mechanism underlying the CSBE-induced mitochondrial apoptotic pathway, in order to investigate the anti-tumor effects of this plant extract. CSBE-induced apoptosis of the SGC-7901 human gastric cancer cell line was observed, and alterations in the expression levels and localization of initiators, markers, and executors of the mitochondrial apoptosis pathway were analyzed. Following treatment of SGC-7901 cells with CBSE, proliferation was inhibited and apoptosis was induced; and these effects were associated with mitochondrial membrane potential disruption, cytochrome c release into the cytoplasm, and caspase-9 and caspase-3 activation. CSBE may have induced SGC-7901 cell apoptosis by upregulating the expression of B-cell lymphoma-2 (BCL-2)-associated X protein, and downregulating the expression of BCL-2. The results of the present study suggested that CSBE may induce SGC-7901 cell apoptosis via activation of the mitochondrial apoptosis pathway.

To identify the estrogenic constituents in Tu-Si-Zi extracts. Estrogen activity of Tu-Si-Zi extracts was assayed by the method of uterotrophic assay and MTT. Chemical ingredients in the serum were confirmed by HPLC/MS-Q-TOF. Ethanol (95%, v/v) extract of Tu-Si-Zi could significantly increase the uterine coefficient of minor mice and the proliferation of MCF-7 cell lines at the dosage of 24 g · kg−1 · d−1(p

Hereditary nonsyndromic hearing loss is extremely heterogeneous. Mutations in the transmembrane channel-like gene1 (TMC1) are known to cause autosomal dominant and recessive forms of nonsyndromic hearing loss linked to the loci of DFNA36 and DFNB7/11, respectively. We characterized a six-generation Chinese family (5315) with progressive, postlingual autosomal dominant nonsyndromic hearing loss (ADNSHL). By combining targeted capture of 82 known deafness genes, next-generation sequencing and bioinformatic analysis, we identified TMC1 c.1714G>A (p. D572N) as the disease-causing mutation. This mutation co-segregated with hearing loss in other family members and was not detected in 308 normal controls. In order to determine the prevalence of TMC1 c.1714G>A in Chinese ADNSHL families, we used DNA samples from 67 ADNSHL families with sloping audiogram and identified two families carry this mutation. To determine whether it arose from a common ancestor, we analyzed nine STR markers. Our results indicated that TMC1 c.1714G>A (p.D572N) account for about 4.4% (3/68) of ADNSHL in the Chinese population.

Chemical examination of the fermentation broth of a sponge-associated fungus Trichoderma harzinum HMS-15-3 led to the isolation of four pairs of new C13 lipid enantiomers namely harzianumols A-H (1a-4b). Their structures were elucidated on the basis of extensive spectroscopic (IR, MS, 1D, and 2D NMR) data analysis, including the modified Mosher's method for the assignment of their absolute configurations. The new compounds were evaluated for antihyperlipidemic effects in HepG2 cells.

Ethnopharmacological relevance Gegenqinlian Decoction (GQD) has been used as a folk remedy for gastrointestinal diseases in China over thousands of years. It has significant treatment efficacy for patients with inflammatory bowel disease (IBD). We analyzed and showed that the active components alignment of Gegenqinlian Decoction (ACAG) possesses broad pharmacological effects including analgesic, antipyretic, anti-inflammatory, antibacterial, antiviral and antidiarrhea, as well as the effect of adjusting gastrointestinal function in our preliminary experiments. However, the exact molecular mechanisms on how ACAG exerts these pharmacological effects still remain elusive. In the present study, the plausible pharmacological effects of ACAG on 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis were investigated. Materials and methods Male Sprague-Dawley (SD) rats with TNBS/ethanol-induced colitis were used. The colonic wet weight, macroscopic and histological colon injury, superoxide dismutase (SOD), malonyldialdehyde (MDA), and inducible nitric oxide synthase (iNOS) activity were observed. Pro-inflammation cytokines were determined by ELISA methods, semi-quantitative RT-PCR and Immuno-histochemistry. Results We showed administration of ACAG was able to improve colitis. This was manifested by a decreased in the score of macroscopic and histological colonic injury, by lowered colonic wet weight, accompanied by significant increased of SOD activity, and decreased of MDA and iNOS activities. The treatment also significantly reduced tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1β) levels in colon and serum as well as the colonic mRNA levels for several inflammatory cytokines such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), macrophage inflammatory protein-2 (MIP-2), intercellular adhesion molecule-1 (ICAM-1) and toll-like receptor 2, 4 (TLR2, TLR4). In addition, we also showed that ACAG was able to inhibit the activation and translocation of transcription factors, nuclear factor kappaBp65 (NF-κBp65) in colon. Conclusions Our results suggest that ACAG exhibits protective effect in TNBS-induced ulcerative colitis. We postulate that this might be due to its modulation of oxidant/anti-oxidant balance, downregulation of productions, expressions of pro-inflammatory cytokines and inhibition of NF-κBp65 signal transduction pathways.

Background Both alcohol and aqueous extracts of Sauromatum giganteum(Engl.) Cusimano and Hett, the dried root tuber of which is named Baifuzi in Chinese, have been used for folklore treatment of cancer in Northeast of China. However, little is known about which is most suitable to the cancer therapy. Materials and methods Serum pharmacology and MTT assays were adopted to detect the effects of ethanol and aqueous extracts of Sauromatum giganteum(Engl.) Cusimano and Hett , prepared by heat reflux methods, on proliferation of different cancer cells. Results Cancer cells treated with medium supplemented with 10%, 20%, 40% serum(v/v) containing ethanol extract had a decline in viability, with inhibition rates of 7.69%, 21.8%, 41.9% in MCF-7 cells, 42.8%, 48.1%, 51.8% in SGC-7901 cells, 44.1%, 49.2%, 53.7% in SMMC-7721 cells, 6.8%, 15.2%, 39.8% in HepG2 cells, 7.57%, 16.3%, 36.2% in HeLa cells, 6.24%, 12.5%, 27.4% in A549 cells, and 7.20%, 17.5%, 31.3% in MDA-MB-231 cells, respectively. Viability in the aqueous extract groups was no different with that of controls. Conclusions An ethanol extract of Sauromatum giganteum(Engl.) Cusimano and Hett inhibited the proliferation of SMMC-7721, SGC-7901 and MCF-7 cells, which supports the use of alcoholic but not aqueous extracts for control of sensive cancers, which might include hepatocarcinoma, gastric cancer and breast cancer.

Trace chemical constituents from the ethyl acetate extract of Red Yeast Rice were investigated. Four phenolic compounds were isolated by various column chromatographies, and their structures were identified on the basis of spectroscopic analysis including UV, MS, IR and NMR. The four compounds were identified as 2-methyl-5-(2'R-methyl-4'-hydroxy-butyl)-cinnamic acid(1), 5-(2'-hydroxy-6'-methyl phenyl)-3-methylfuran-2-carboxylic acid(2), daidzein(3), and genistein(4). Compound 1 was new and 2 was firstly discovered from the genus Monascus, while 3-4 were obtained from Red Yeast Rice for the first time.

Compared with chemical drugs, it is a huge challenge to identify active ingredients of multicomponent traditional Chinese medicine (TCM). For most TCMs, metabolism investigation of absorbed constituents is a feasible way to clarify the active material basis. Although Andrographis paniculata (AP) has been extensively researched by domestic and foreign scholars, its metabolism has seldom been fully addressed to date. In this paper, high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry was applied to analysis and characterization of AP metabolism in rat urine and feces samples after oral administration of ethanol extract. The differences in metabolites and metabolic pathways between the two biological samples were further compared. The chemical structures of 20 components were tentatively identified from drug-treated biological samples, including six prototype components and 14 metabolites, which underwent such main metabolic pathways as hydrolyzation, hydrogenation, dehydroxylation, deoxygenation, methylation, glucuronidation, sulfonation and sulfation. Two co-existing components were found in urine and feces samples, suggesting that some ingredients' metabolic processes were not unique. This study provides a comprehensive report on the metabolism of AP in rats, which will be helpful for understanding its mechanism.

Background The mutation frequencies of three common deafness genes (MT-RNR1 m.1555A>G, GJB2, and SLC26A4) among patients with nonsyndromic sensorineural hearing loss (NSHL) were different in previous studies. Inconsistent selection criteria for recruiting patients could have led to differences in estimating the frequencies of genetic mutations thus resulting in different mutation frequencies among these studies. The aim of this study was to reveal the differences in the mutation spectrums of the three common genes between familial and sporadic Chinese Han patients. Methods Totally, 301 familial probands and 703 sporadic patients with NSHL were enrolled in this study. Three genes, MT-RNR1 m.1555A>G, GJB2, and SLC26A4, were screened for mutation in our study cohort. A χ(2) test was performed to compare the mutation frequencies between the two groups. Results The study showed that the disease-causing mutation frequencies of MT-RNR1 m.1555A>G, GJB2, and SLC26A4 were 12.29%, 14.62%, and 18.27% in familial probands and 3.56%, 18.63%, and 18.92% in sporadic patients, respectively. The mutation frequency of MT-RNR1 m.1555A>G in familial probands was significantly higher than in sporadic patients (χ(2) test, P = 0.000), while there were no significant differences in the mutation frequencies of GJB2 and SLC26A4 between the familial and sporadic groups (χ(2) test, P > 0.05). Conclusions It is necessary to reveal the differences in gene mutation frequencies between patients of different sources or characteristics by comparative studies in order to avoid selection bias. The mutations of GJB2, SLC26A4, and MT-RNR1 m.1555A>G are the most important etiological factors in Chinese Han patients, among which SLC26A4 might be the most frequent.

Hepatocellular carcinoma (HCC) has a relatively higher incidence in many countries of Asia. Globally, HCC has a high fatality rate and short survival. Epirubicin, a doxorubicin analogue, may be administered alone or in combination with other agents to treat primary liver cancer and metastatic diseases. However, the toxic effects of epirubicin to normal tissues and cells have been one of the major obstacles to successful cancer chemotherapy. Here, we investigated the effects of epirubicin in combination with kappa-selenocarrageenan on mice with H22 implanted tumors and HepG-2 cell proliferation, immune organ index, morphology, cell cycle and related protein expressions in vivo and in vitro with sequential drug exposure. The inhibitory rate of tumor growth in vivo was calculated. Drug sensitivity was measured by MTT assay, and the King's principle was used to evaluate the interaction of drug combination. Morphological changes were observed by fluorescent microscopy. Cell cycle changes were analyzed by flow cytometry. Expression of cyclin A, Cdc25A and Cdk2 were detected by Western blotting. In vivo results demonstrated that the inhibitory rate of EPI combined with KSC was higher than that of KSC or EPI alone, and the Q value indicated an additive effect. In addition, KSC could significantly raise the thymus and spleen indices of mice with H22 implanted tumors. In the drug sensitivity assay in vitro, exposure to KSC and EPI simultaneously was more effective than exposure sequentially in HepG-2 cells, while exposure to KSC prior to EPI was more effective than exposure to EPI prior to KSC. Q values showed an additive effect in the simultaneous group and antagonistic effects in the sequential groups. Morphological analysis showed similar results to the drug sensitivity assay. Cell cycle analysis revealed that exposure to KSC or EPI alone arrested the cells in S phase in HepG-2 cells, exposure to KSC and EPI simultaneously caused accumulation in the S phase, an effect caused by either KSC or EPI. Expression of cyclin A, Cdc25A and Cdk2 protein was down-regulated following exposure to KSC and EPI alone or in combination, exposure to KSC and EPI simultaneously resulting in the lowest values. Taken together, our findings suggest that KSC in combination with EPI might have potential as a new therapeutic regimen against HCC.

A number of scientific studies have revealed that laminarin has antitumor effects. Therefore, the aim of the present study was to investigate the apoptosis of LoVo cells and the underlying mechanisms induced by laminarin. LoVo cells were treated with various concentrations of laminarin and fluorescence-inverted microscopy was used to observe the morphology of LoVo cells treated with laminarin. In addition, western blotting was performed to analyze the expression levels of death receptor (DR)4, DR5, TNF-related apoptosis-inducing ligand (TRAIL), Fas-associated protein with death domain (FADD), caspase-8, caspase-3, Bid and tBid. Flow cytometry was conducted to analyze the expressions of Bcl-2 and Bax, and spectrophotometry was performed to quantify the activity of caspases-8, -3, -6 and -7. Following the treatment of LoVo cells with laminarin for 24 h, the expression levels of DR4, DR5, TRAIL, FADD, Bid, tBid and Bax were observed to be upregulated, whereas the expression levels of pro-caspase-8, pro-caspase-3 and Bcl-2 were downregulated. In addition, the activities of casapse-8, -3, -6 and -7 were observed to increase, which was a significant difference when compared with those of the control group. Therefore, laminarin is considered to induce the apoptosis of LoVo cells, which may occur via a DR pathway, suggesting that laminarin may be a potent agent for cancer treatment.

Numerous studies have demonstrated that podophyllotoxin and its derivatives exhibit antitumor effects. The aim of the present study was to investigate SGC-7901 cell apoptosis and the underlying mechanism induced by podophyllotoxin. SGC-7901 cells were treated with varying concentrations of podophyllotoxin. MTT assays and flow cytometry were used to evaluate the effects of podophyllotoxin on the proliferation and apoptosis of SGC-7901 cells, while fluorescence inverted microscopy was used to observe the morphology of SGC-7901 cells that had been dyed with Hoechst 33258. In addition, laser scanning confocal microscopy was used to analyze the mitochondrial membrane potential (MMP) of SGC-7901 cells dyed with Rhodamine 123. Western blotting was performed to analyze the expression levels of cytochrome c (cyt-c), caspase-9 and caspase-3 in the SGC-7901 cells. The results indicated that podophyllotoxin was capable of inhibiting growth and inducing the apoptosis of SGC-7901 cells in a dose-dependent manner, causing cell cycle arrest at the G2/M phase. After 48 h of treatment, the apoptotic morphology of SGC-7901 cells was clear, exhibiting cell protuberance, concentrated cytoplasms and apoptotic bodies. Following 24 h of treatment, the MMP of the SGC-7901 cells decreased. In addition, after 48 h, the expression of cyt-c was shown to be upregulated, while the expression levels of pro-caspase-9 and pro-caspase-3 in the SGC-7901 cells were shown to be downregulated. In conclusion, apoptosis can be induced in SGC-7901 cells by podophyllotoxin, potentially via a mitochondrial pathway, indicating that podophyllotoxin may be a potent agent for cancer treatment.

Background: Cistanche deserticola, a traditional Chinese herb medicine, has been widely used for thousands of years with the activities of hormone regulation, immunomodulatory, antioxidative, neuroprotective, anti-inflammatory, and estrogen. Glycosides of Cistanches (GCs) were the main bioactivity components of the herb. Objective: The objective of the study is to study estrogenic activity and the mechanism about estrogen receptors (ERs) of GCs. Materials and Methods: Cell proliferation was measured using the 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyltetrazolium bromide assay for MCF-7 cells. The cell cycle was detected using flow cytometry, and proliferation index was calculated. The mRNA and protein expressions of ERα and ERβ were detected by reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis as the reported method with minor modifications. Results: GCs group at the concentrations of 1.75, 17.5, and 175 μg/mg could enhance proliferation of the MCF-7 cell lines with a time and dosage-dependent manner. Combined medication group (fulvestrant with estradiol [E2] or GCs) could lead to the incline of proliferation rate compared with the individual medication group (P

The purpose of this study is to investigate the extraction and separation of alkaloids from H.littoralis. Chopped bulbs of H.littoralis was soaked in a mixture of dichloromethane and methanol (1:1) for 24 h, ultrasonic extraction six times, then extracted with N-butanol to yield crude extracts. Three components were isolated from N-butanol extract of H.littoralis by Sephadex LH-20 and TLC. The compounds were identified by LC-MS, they are 5,6-dihydrobicolorine, Lycorine and 7-deoxy-trans-dihydronarciclasine.

The basic structures of Crinum Asiaticum alkaloids are N-(3, 4-dioxo-benzyl)-4-O-phenylethylamine, pyrrole phenanthridine, 5,10b-ethanophenanthridine. The basic framework can form a new parent nucleus structure with significant pharmacological activity by oxidation, dehydrogenation, cyclization, O, N-methylation, acylation reaction. Crinum Asiaticum alkaloids antibacterial, antimalarial, anti-inflammatory, anti-cancer, inducing tumor cell apoptosis, the protection of the cardiovascular and other effects are reviewed.

The purpose of this study is to investigate the effect of fucosterol on the induction of apoptosis and the molecular mechanism involved in Human promyelocytic leukemia HL-60 Cells. HL-60 Cells were treated with different concentrations of fucosterol at different time. MTT method was used to study fucosterol anti-tumor activity. Morphology observation was performed to determine the effects of fucosterol on apoptosis of HL-60 cells. Flow cytometry (FCM) was used to detect the cell cycle. Laser scanning confocal microscope (LSCM) was used to analyze the expressions of Fas, FasL, Fadd and Caspase-8. Caspase activity kits were used to determine the activity of Caspase-8 and Caspase-3. The results showed fucosterol could inhibit the growth of HL-60 cells, and the apoptosis morphology for 48 h treatment was obvious, which showed cell protuberance, cytoplasm concentrated and apoptotic body. Fucosterol treatment for 24 h increased the protein expression of Fas, FasL, Fadd and Caspase-8. It also showed that the activity of Caspase-3 and Caspase-8 has increased significantly. In conclusion, Fucosterol could induce HL-60 cells apoptosis via death receptor pathway.

The present paper reviews the research on the medicinal value of Amaryllidaceae plants. The medicinal value of Amaryllidaceae plants attracts much attention nowadays by By discovery the pharmaco-activity of alkaloid series within plants etc. Lycoris radiata, Crinum asiaticum L. Kafir Lily, Curculigo orchioides gaertn. This paper mainly discourses the mecicinal value on the pharmaco-activity of Amaryllidaceae plants. The study found that Amaryllidaceae plants have effects on anti-tumor, antiinflammatory, antibiosis, and antiviral, antimalarial and so on, and have some effects on the immune system, central nervous system, inhibition AcCHE.

socarbostyril alkaloids is a kind of alkaloid does not contain basic nitrogen atoms and is represented by hydroxylated benzophenanthridone or isoquinolinone types of structure. The most widely known compounds of this group are narciclasine, lycoricidine , and pancratistatin. They have demonstrated to inhibite the proliferation of many human cancer cells, and at the same time have no affect on normal human cells under a certain dose, they have a high efficiency and low toxicity in antitumor area. Now this kind of compound has been a hot spot research to antitumor workers. The present paper reviews the origin and the antitumor function of the Isocarbostyril alkaloids.

Lycorine is a kind of natural alkaloid extracted from Amaryllidaceae that has shown various pharmacological effects. Recent studies have focused on the potential antitumor activity of lycorine. The inhibition effects of lycorine on the cell proliferation of leukemia HL-60 cells, K562 cells, chronic lymphocytic leukemia, U937 cells and structure-activity studies on the lycorine pharmacophore in leukemia cells were summarized in the article. The review of pharmacological effects and structure-activity on leukemia of lycorine can provide a reference for the further development and utilization on the kind of natural products.

The development of tumor is not only results of the uncontrolled cell proliferation, the differentiation and cell abnormalities, but also relates to apoptosis imbalance. Inducing tumor cell apoptosis has become an important means of clinical applications on cancer treatment. Apoptosis of tumor cells is affected by a variety of gene regulation, including some oncogenes and tumor suppressor genes which are related to cell proliferation. Focusing on the three basic ways of apoptosis: death receptor pathway, the mitochondrial pathway, and endoplasmic reticulum stress pathway, This paper summarizes the research progress of appoptosis factors for the past few years and provides information and ideas for finding out effective anticancer drugs which have key anti-cancer factors as target points.

The purpose of this study is to investigate the effect of fucosterol on the induction of apoptosis and the molecular mechanism involved in Human promyelocytic leukemia HL-60 Cells. HL-60 Cells were treated with different concentrations of fucosterol at different time. MTT method was used to study fucosterol anti-tumor activity. Morphology observation was performed to determine the effects of fucosterol on apoptosis of HL-60 cells. Flow cytometry (FCM) was used to detect the cell cycle. Laser scanning confocal microscope (LSCM) was used to analyze mitochondrial membrane potential (MMP). Western blot was performed to analyze the expressions of Cyt-C, Caspase-9 and Caspase-3. The results showed fucosterol could inhibit the growth of HL-60 cells, and the apoptosis morphology for 48 h treatment was obvious, which showed cell protuberance, cytoplasm concentrated and apoptotic body. Fucosterol treatment for 24 h decreased MMP in dose-dependent manners. It also induced the release of Cyt-C and the activation of Caspase-9 and-3. In conclusion, Fucosterol could induce HL-60 cells apoptosis through a mitochondrial pathway.

This paper investigated the selenylation, characterization and cytotoxic activity of seleno-Astragalus polysaccharide. Firstly, Se-APS was synthesized under reaction time at 8 h, reaction temperature at 80 °C, ratio of Na2SeO3 to APS at 1.0 g/g and water bath shaking rate at 40 r/min. Then, a series of experiments were designed to investigate the characterization and cytotoxicity of Se-APS. The result indicated that the characterization of Se-APS was significantly different from APS, except for X-ray diffraction. Additionally, MTT assay conformed that Se-APS could significantly inhibit the proliferation of human breast cancer MCF-7 cells in dose-dependent and time-dependent manners.