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Systemic lupus erythematosus (SLE) is a chronic, devastating autoimmune disorder associated with severe organ damage. The roles of Toll-like receptor 9 (TLR9) and NETosis in SLE have been described, suggesting the involvement of NETosis signaling in the development of SLE. Shaoyao-Gancao Decoction (SGT) is a potential medication for the treatment of SLE; however, its potential therapeutic mechanism remains unexplored. To determine the function of SGT in SLE, we treated MRL/lpr female mice with SGT, the main components of which were paeoniflorin (56.949 μg·mL−1) and glycyrrhizin (459.393 μg·mL−1). We found that SGT treatment relieved lymphadenectasis and splenomegaly, reduced urine protein and anti-dsDNA antibody concentrations, and relieved kidney pathology in MRL/lpr mice. SGT could also effectively regulate the oxidation/antioxidant balance, significantly reduce malondialdehyde (MDA) and nitric oxide (NO) contents and significantly increase superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in MRL/lpr mice. The neutrophil extracellular trap (NET) content of MRL/lpr mice also decreased to a certain extent after SGT treatment. All these results suggested that SGT might improve the inflammatory damage to tissues caused by oxygen free radicals, thereby regulating the NETosis process mediated by TLR9 and exerting a good therapeutic effect on SLE.

The correlation between dietary inflammation index (DII) and rheumatoid arthritis (RA) has been found, but the effect of confounding factors is not considered. This study aims to further explore the association between DII and RA risk by taking the Americans as the research object.The data from the 2005-2018 National Health and Nutrition Examination Survey (NHANES) database included 1819 self-reported RA individuals and 8602 non-RA individuals. The analytical methods include logistic regression, additive model, smooth curve fitting, and the recursive algorithm.There was a positive correlation between DII and RA in Americans (β = 1.068, 95% CI = 1.026 to 1.111, P = 0.001). This result was still presented in the subgroup analysis, including age less than 50 years, female, other Hispanics, BMI ≥ 25, and federal poverty rate 185%, and it was more pronounced in smokers. The results show that the superposition of DII and other risk factors would increase the risk of RA (β 1.068). In addition, individuals with RA are inadequate in intake of anti-inflammatory foods, in line with the Mediterranean diet.The inflammatory potential of the diet is positively correlated with the risk of RA, and has a superimposed effect with other risk factors, increasing the probability of the risk of disease. These results emphasize that reducing the intake of pro-inflammatory foods may be an effective measure to prevent the onset of rheumatoid arthritis. However, eating anti-inflammatory foods exclusively is not the best option. Intaking some pro-inflammatory foods like protein, energy, and total saturated acids may be necessary to maintain the physiological function of the human body. Key Points • Dietary inflammation index (DII) is positively correlated with RA risk. • When DII and other risk factors appear at the same time, the effects of the two will be superimposed on each other, increasing the risk of RA. • When the DII is the same, Hispanic has a higher incidence of RA. • Among the pro-inflammatory foods, the intake of protein, energy, and saturated fatty acids is still required by RA patients.

A correlation between mental illness and systemic rheumatoid arthritis (RA) has been observed in several prior investigations. However, little is known about the causative relationship between them. The present study aimed to systematically investigate the potential association between genetically determined mental illness and RA. Two-sample bidirectional Mendelian randomization (MR) analysis was performed using publicly released genome-wide association studies (GWAS). We selected independent genetic variants associated with four mental illnesses (bipolar disorder, broad depression, major depression, and anxiety) as instrumental variables. The inverse variance weighted (IVW) method was used as the primary analysis to assess the causal relationship between mental illness and RA. Results of the IVW analysis suggested that genetic predisposition to bipolar disorder was associated with a decreased risk of RA (odds ratio [OR] = 0.825, 95% CI = 0.716 to 0.95, p = 0.007). Furthermore, we did not find a significant causal effect of RA on bipolar disorder in the reverse MR analysis (p > 0.05). In addition, our study found no evidence of a bidirectional causal relationship between genetically predicted broad depression, major depression, anxiety, and RA (p > 0.05). The genetically proxied bipolar disorder population has a lower RA risk, which may indicate that there is a hidden mechanism for inhibiting the pathogenesis of RA in bipolar disorder. However, results do not support a causal connection between depression, anxiety, and RA.

ObjectiveRecent studies reported that SLE is characterised by altered interactions between the microbiome and immune system. We performed a meta-analysis of publications on this topic.MethodsCase–control studies that compared patients with SLE and healthy controls (HCs) and determined the diversity of the gut microbiota and the abundance of different microbes were examined. Stata/MP V.16 was used for the meta-analysis. A Bonferroni correction for multiple tests was used to reduce the likelihood of false-positive results.ResultsWe included 11 case–control studies that examined 373 patients with SLE and 1288 HCs. These studies were performed in five countries and nine cities. Compared with HCs, patients with SLE had gut microbiota with lower Shannon-Wiener diversity index (weighted mean difference=−0.22, 95% CI −0.32 to –0.13, p0.05). Patients with SLE who used high doses of glucocorticoids had altered gut microbiota based on the Chao1 species diversity estimator, and hydroxychloroquine use appeared to reduce the abundance of Enterobacteriaceae.ConclusionsPatients with SLE have imbalanced gut microbiota, with a decrease in beneficial bacteria and an increase in harmful bacteria. Drugs used to treat SLE may also alter the gut microbiota of these patients.

Development of a fully functional dressing that can be applied throughout wound healing remains challenging. In this study, Bletilla striata polysaccharide (BSP) and the ethanol extract of B. striata (EEB) were physically blended and vacuum freeze-dried into a new medical dressing with various functions, such as hemostasis, anti-inflammatory, antibacterial and wound healing promotion. The in vivo and in vitro blood coagulation evaluation results showed that the most obvious hemostasis effects coming from the EEB25 sponge was probably due to good water absorption capacity. The drug sensitivity test results showed that the sponges presented a good antibacterial effect originated from the addition of EEB. In addition, the EEB25 sponge showed significant anti-inflammatory effects, as well as excellent biodegradability and biocompatibility. The full-thickness wound experiment illustrated the wound healing rates of EEB25 group on the 8th day and 16th day was 82.76 ± 2.78% and 95.36 ± 3.72%, respectively, which showed significant differences (p

Introduction: Evaluate the changes of gut Microbiota in patients with Systemic Lupus Erythematosus (SLE) and healthy people by meta-analysis. Methods We searched the case-control studies of SLE and healthy controls (HCs) for detecting the diversity of gut Microbiota and the abundance level of some microbiota in the two groups. StataMP16 software was applied for this meta-analysis. The Newcastle-Ottawa quality assessment scale (NOS) was used to assess the quality of the included studies. Results Eleven case-control studies were included. There were 373 SLE patients and 1288 healthy people, involving 5 countries and 9 different cities. Compared with the HCs, the Shannon-wiener diversity index (WMD=-0.22; 95% CI=-0.32 to -0.13; P = 0.000) and Chao1 richness estimator (SMD=-0.62; 95% CI=-1.04 to -0.21; P = 0.003) of gut Microbiota in SLE decreased, and the abundance level of Ruminococcaceae decreased (SMD=--0.48; 95% CI = 0.76 to-0.21; P = 0.001). Enterobacteriaceae (SMD = 0.39,95% CI = 0.11 to 0.66;P = 0.006) and Enterococcaceae (SMD = 0.55; 95% CI = 0.19 to 0.9; P = 0.03) showed higher abundance levels in comparison with HCs. The subgroup analysis showed the abundance level of Ruminococcaceae (SMD=-0.89; 95% CI =-1.34 to -0.45; P = 0.000) was lower and Enterococcaceae was higher (SMD = 0.77; 95% CI = 0.34 to 1.21༛P = 0.001) in Chinese with SLE compared with HCs. In non-Chinese patients with SLE, there were no significant difference between the abundance level of Ruminococcaceae (SMD=-0.22; 95% CI=--0.58 to 0.13; P = 0.216) and Enterococcaceae (SMD=-0.08; 95% CI=-0.49 to 0.32; P = 0.682 ) with HCs. The subgroup analysis also found the level of Enterobacteriaceae was affected by the sample size. Conclusion Compared with the diversity of healthy people, richness and evenness of gut microbiota in patients with SLE are impaired. There is a decrease in the abundance level of beneficial bacteria and an increase in the harmful bacteria. Thus, gut microbiota in patients with SLE appear disorder, which may lead to metabolic imbalance, destruction of the integrity of the small intestine, immune system disorders and pro-inflammatory. Regulating the abundance of gut microbiota can be used as one of the key strategies for treating SLE.

Jieduquyuziyin prescription (JP) is a traditional Chinese medicinal (TCM) formula which has been demonstrated to be effective for systemic lupus erythematosus (SLE) treatment as an approved hospital prescription for many years in China. But its mechanism of action in combating this disease is largely unknown. Our previous studies showed that JP can slow disease progression without producing significant toxic side effects. We treated MRL/lpr mice with JP to ascertain if JP could improve SLE by the suppression of Ca(2+)/calmodulin-dependent protein kinase-4 (CaMK4) expression. We investigated the role of JP in a model of SLE in MRL/lpr mice, and identified the possible mechanism of action. Mice were divided randomly into four groups: control, model, and two treatment groups. Sections of renal tissue were stained with hematoxylin and eosin (HE). Histopathologic changes in the kidney were evaluated by light microscopy. T-helper (Th)17 cells were analyzed by flow cytometry. mRNA levels of interleukin (IL)-17, CaMK4, and RORγt were measured by reverse transcription polymerase chain reaction (RT-PCR). CaMK4 expression was assessed by Western blotting. The results showed that the percentages of Th17, IL-17, and RORγt in mice treated with JP were decreased remarkably compared to the model group (p 0.05). Interestingly, a high CaMK4 expression was observed in the SLE mice, which was inhibited by JP. These results suggest that CaMK4 activity was increased in T cells from MRL/lpr mice compared with the control group. Our findings support the conclusion that the effects of JP on MRL/lpr mice may involve the regulation of CaMK4 overexpression in MRL/lpr mice.

Cardamonin has promising potential in cancer prevention and therapy by interacting with proteins and modifying the expressions and activities, including factors of cell survival, proliferation, and angiogenesis. In our precious study, we have demonstrated that cardamonin suppressed vascular endothelial growth factor- (VEGF-) induced angiogenesis as evaluated in the mouse aortic ring assay. It is also known that microRNAs (miRNAs) play important roles in angiogenesis. Herein, we hypothesized whether antiangiogenesis effect of cardamonin in human umbilical vein endothelial cells (HUVECs) triggered by VEGF was associated with miRNAs. We found that cardamonin reduced the miR-21 expression induced by VEGF in HUVECs. Treatment with miR-21 mimics abolished the effects of cardamonin on VEGF-induced cell proliferation, migration, and angiogenesis in HUVECs. However, treatment with miR-21 inhibitors presented the opposite effects, indicating the vital role of miR-21 in this process. Our study provides a new insight of the preliminary mechanism of anti-VEGF-induced angiogenesis by cardamonin in HUVECs.

A urine metabolomics method based on gas chromatography-mass spectrometry was developed in order to investigate the metabolite characteristics of systemic lupus erythematosus (SLE) and the therapeutic effects of Jieduquyuziyin prescription. The urinary metabolic profiles in urine specimens of the SLE model mice (MRL/lpr) group, prednisone acetate-treated SLE mouse group, Jieduquyuziyin prescription-treated SLE mouse group, and control group (C57BL/6 J) after the administration were analyzed by gas chromatography-mass spectrometry. These metabolic profiles were then processed by multivariate analysis, in particular Mass Profiler Professional, SIMCA-P and partial least-squares discriminant analysis. According to the partial least-squares discriminant analysis results, the SLE model group and the control group were obviously separated, indicating that the incidence of SLE had a greater impact on the metabolic network, and the SLE model group had significant difference compared with the control group in urine metabolites. Eleven differential metabolites were identified to be related to SLE, and the results of differential metabolite identification showed that the metabolites were mainly related to energy metabolism and amino acid metabolism pathway. These results can provide an experimental basis for further exploring the mechanism of traditional Chinese medicine in the treatment of SLE.

A serum metabolomics method based on rapid resolution liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (RRLC-Q-TOF-MS) was performed for a holistic evaluation of the metabolic changes of collagen-induced arthritis (CIA) in rats and to assess the interventional effects of type II collagen (CII) in this model. Partial least-squares-discriminant analysis (PLS-DA) was employed to study the metabolic profiling of CIA rats and control rats. Ten metabolites, namely, 12(S)-HHTrE, 12(S)-HEPE, PGE2, TXB2, 12(S)-HETE, LysoPE(16:0), PE(O-18:0/0:0), Lyso-PE(18:2), Lyso-PE(20:4), and Lyso-PC(22:5) were identified as differential metabolites associated with the pathogenesis of CIA. These results suggested that dysregulation of the arachidonic acid (AA) and phospholipid metabolic networks is involved in the pathomechanism of CIA. Differential metabolomics and histopathological analyses demonstrated that CII inhibits the progress of arthritis. Furthermore, the therapeutic effects of CII on CIA may involve regulation of the disordered AA and phospholipid metabolic networks. This metabolomics study provides new insights into the pathogenesis of arthritis and, furthermore, indicates the potential mechanism underlying the significantly increased prevalence of metabolic syndrome, defined as a clustering of cardiovascular disease (CVD) risk factors, in arthritis patients.

A metabolomics approach was applied to study the metabolites characters of systemic lupus erythematosus (SLE) and therapeutical effects of jieduquyuziyin prescription (JP). High performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF/MS) in positive mode was employed to analyze urine specimens of the SLE model mice (MRL/lpr) group, prednisone acetate (PA)-treated SLE mice group, JP-treated SLE mice group, and control group (C57BL/6 J). Software of Mass Hunter and Mass Profiler Professional (MPP) were used to process the data. Multivariate statistical analysis of principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) were applied to recognize the data pattern. PCA result showed that there were obvious disorders of amino acid metabolism in SLE model. Ten compounds have significant contributions to classification were identified as differential metabolites, such as glycine, pantetheine, and phenylpyruvic acid. Metabolism of phenylalanine tended to recover to the normal state after treatment with PA. The changes of glycine, phenylalanine, and tryptophan in SLE mice showed some alleviation in JP-treated SLE group. Thus, the JP has a good therapeutic effect on the SLE model mice by regulating the dysfunctions of amino acid metabolism. These results may be helpful to further understand the pathological mechanisms of SLE from the whole metabolism.

Cardiovascular disease is one of the most serious diseases threatening human health. Puerarin has been widely used in china as a cardiovascular agent. However, a major clinical limitation is its adverse effects of hemolysis. We have addressed this issue by PEGylation and synthesized a puerarin prodrug by covalently linking puerarin to the drug carrier monomethoxy-poly(ethylene glycol) (mPEG). The covalent linkage was validated by UV, FTIR, 1H-NMR, and matrix-assisted laser desorption ionization-time-of-flight mass spectrometry. The hemolysis assay showed that the prodrug produced minimal hematotoxicity. The hydrolysis profile showed that the prodrug was stable in a low pH environment and can continuously release the active drug in physiological condition in vitro. A pharmacokinetic study showed a prolongation of the elimination half-life and a favorable distribution profile in organs, suggesting that the prodrug preparation produced fewer side effects on liver than the injectable form of puerarin normally used. Additionally, the protective effect of the prodrug on acute myocardial ischemia model implied better therapeutic effect than injectable puerarin. We show that the puerarin PEG conjugation is a promising prodrug that produces fewer side effects, exhibits better aqueous solubility, and is more effective than puerarin. © 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013

The interaction between bovine serum albumin (BSA) and pegylated puerarin (Pur) in aqueous solution was investigated by UV-Vis spectroscopy, fluorescence spectroscopy and circular dichroism spectra (CD), as well as dynamic light scattering (DLS). The fluorescence of BSA was strongly quenched by the binding of pegylated Pur to BSA. The binding constants and the number of binding sites of mPEG 5000-Pur with BSA were 2.67 ± 0.12 and 1.37 ± 0.05 folds larger after pegylating, which were calculated from the data obtained from fluorescence quenching experiments. The enthalpy change (ΔH) and entropy change (ΔS) were calculated to be 4.09 kJ mol -1 and 20.01 J mol -1 K -1, respectively, according to Van't Hoff equation, indicating that the hydrophobic force plays a main role in the binding interaction between pegylated Pur and BSA. In addition, the negative sign for Gibbs free energy change (ΔG) implies that the interaction process is spontaneous. Moreover, the results of synchronous fluorescence and CD spectra demonstrated that the microenvironment and the secondary conformation of BSA were changed. Comparing with Pur, all our data collected indicated that pegylated Pur interacted with BSA in the same way as that of Pur, but docked into the hydrophobic pocket of BSA with more accessibility and stronger binding force. DLS measurements showed monomethoxy polyethylene glycol (mPEG) have an effect on BSA conformation, and revealed that changes in BSA size might be due to increases in binding constant and the absolute values of ΔG after Pur pegylation. © 2011 Elsevier B.V.

Two peptides of Pt-A (Glu-Asn-Trp 429 Da) and Pt-B (Glu-Gln-Trp 443 Da) were isolated from venom liquor ofDeinagkistrodon acutus. Their antiplatelet aggregation effects were evaluated with platelet-rich human plasmain vitro; the respective IC50of Pt-A and Pt-B was 66 μM and 203 μM. Both peptides exhibited protection effects on ADP-induced paralysis in mice. After ADP administration, the paralysis time of different concentration of Pt-A and Pt-B lasted as the following: 80 mg/kg Pt-B (152.8 ± 57.8 s) < 40 mg/kg Pt-A (163.5 ± 59.8 s) < 20 mg/kg Pt-A (253.5 ± 74.5 s) < 4 mg/kg clopidogrel (a positive control, 254.5 ± 41.97 s) < 40 mg/kg Pt-B (400.8 ± 35.9 s) < 10 mg/kg Pt-A (422.8 ± 55.4 s), all of which were statistically shorter than the saline treatment (666 ± 28 s). Pulmonary tissue biopsy confirmed that Pt-A and Pt-B prevented the formation of thrombi in the lung. Unlike ADP injection alone, which caused significant reduction of peripheral platelet count, Pt-A treatment prevented the drop of peripheral platelet counts; interestingly, Pt-B could not, even though the same amount of Pt-B also showed protection effects on ADP-induced paralysis and thrombosis. More importantly, intravenous injection of Pt-A and Pt-B did not significantly increase the hemorrhage risks as clopidogrel.

The effects of different aeration conditions on xylitol production from corncob hemicellulose hydrolysate by Candida sp. ZU04 were investigated. Batch fermentations were carried out in a 3.7-L fermentor at 30 degrees C, pH 5.5, and agitation of 300 rpm. It was found that the two-phase aeration process was more effective than the one-phase aeration process in xylitol production. In the first 24 h of the aerobic phase, a high aeration rate was applied, glucose was soon consumed, and biomass increased quickly. In the second fermentation phase, aeration rate was reduced and an improved xylitol yield was obtained. The maximum xylitol yield (0.76 g/g) was obtained with an aeration rate of 1.5 vvm (KLa of 37 h-1) for the first 24 h and 0.3 vvm (KLa of 6 h-1) from 24 to 96 h.

Jieduquyuziyin prescription (JP), a traditional Chinese medicine (TCM) prescription, has been widely used for the clinical treatment of systemic lupus erythematosus (SLE). However, the complex chemical constituents of JP and the multifactorial pathogenesis of SLE make research on the therapeutic mechanism of JP in SLE challenging. In this paper, a serum metabolomics approach based on rapid resolution liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (RRLC-Q-TOF/MS) was employed to acquire the metabolic characteristics of serum samples obtained from mice in the SLE model group, JP-treated group, prednisone acetate (PA)-treated group and control group. The orthogonal partial least squares (OPLS) was applied to recognize metabolic patterns, and an obvious separation of groups was obtained. Thirteen metabolites, namely, phosphatidylethanolamine (PE 20:3), hepoxilin B3, lyso- phosphatidylethanolamine (lyso-PE 22:6), 12S-hydroxypentaenoic acid (12S-HEPE), traumatic acid, serotonin, platelet-activating factor (PAF), phosphatidylcholine (PC 20:5),eicosapentaenoic acid (EPA), 12(S)-hydroxyei- cosatetraenoic acid (12S-HETE), 14-hydroxy docosahexaenoic acid (14-HDOHE), lyso-phosphatidylcholine (lyso-PC 20:4), and indole acetaldehyde, were identified and characterized as differential metabolites involved in the pathogenesis of SLE. After treatment with JP, the relative content of 12(S)-HETE, PAF, 12(S)-HEPE, EPA, PE (20:3), Lyso-PE(22:6), and 14-HDOHE were effectively regulated, which suggested that the therapeutic effects of JP on SLE may involve regulating disturbances to the metabolism of unsaturated fatty acid, tryptophan and phospholipid. This research also demonstrated that metabolomics is a powerful tool for researching complex disease mechanisms and evaluating the mechanism of action of TCM.

This study was carried out to evaluate the utilization probability of the fibrous root part (FRP) of Bletilla striata, which was usually discarded and harvesting pseudobulb part (PSP). The chemical composition, total phenolic content, DPPH radical scavenging activity, Ferric-reducing antioxidant power and tyrosinase inhibition activity were compared between FRP and PSP. Antioxidant and pro-oxidant effect as well as antitumor effect of the extract of FRP and PSP were analyzed by in vitro cell system as well. Thin layer chromatography and high performance liquid chromatography analysis indicated that the chemical compositions in the two parts were similar, but the content in FRP was much higher than PSP. Meanwhile, the FRP extracts showed higher phenolic content, stronger DPPH scavenging activity, Ferric-reducing antioxidant capacity and tyrosinase inhibition activity. Sub-fraction analysis revealed that the distribution characteristic of phenolic components and other active constituents in FRP and PSP were consistent, and mainly deposited in chloroform and acetoacetate fractions. Especially, the chloroform sub-fraction (sch) of FRP showed extraordinary DPPH scavenging activity and tyrosinase inhibition activity, with IC50 0.848 mg/L and 4.3 mg/L, respectively. Besides, tyrosinase inhibition activity was even stronger than the positive compound arbutin (31.8 mg/L). Moreover, In vitro cell system analysis confirmed that FRP extract exerts comparable activity with PSP, especially, the sub-fraction sch of FRP showed better antioxidant activity at low dosage and stronger per-oxidant activity at high dosage, and both sch of FRP and PSP can dose-dependent induce HepG2 cells apoptosis, which implied tumor therapeutic effect. Considering that an additional 0.3 kg FRP would be obtained when producing 1.0 kg PSP, our work demonstrated that FRP is very potential to be used together with PSP.

In order to investigate systemic lupus erythematosus (SLE) possible disease biomarkers and disclose its pathogenesis, a rapid resolution liquid chromatography combined with quadrupole-time of flight mass spectrometry method (RRLC-Q-TOF/MS) was used to study the sera from 17 SLE patients and 15 healthy volunteers. Supervised pattern recognition methods combined with orthogonal signal correction partial least square-discriminate analysis (OSC-PLS-DA) were chosen to process the metabolic data. A nice distinction between SLE patients and healthy control group was observed. Further, ten ions, which had a significant contribution to classification, had been selected from metabolic profiles of SLE patients. The ten ions were identified and belong to seven metabolic biomarkers. It was shown that there were abnormal metabolisms of amino acids, phospholipids and porphyrin in SLE patients. It provided a scientific basis for the monitoring and diagnosis of SLE.

Pinellia ternate (Thunb.) Breit (Araceae) is an important Chinese medicinal herb. In order to obtain the hexadecaploid plants of P. ternate with improved medicinal qualities, the shoot tips from in vitro-grown plants were treated with colchicine at various concentrations for different time to double the chromosome number. We obtained highest (42.5%) frequency of morphological variation when the tips were treated with 0.1% colchicine for 48 h. The putative hexadecaploid plants showed characteristics such as larger and thicker leaves, deeper green leaf color, stouter petiole, larger stomata, lower density of stomata across the lower epidermis of leaves and increased numbers of chloroplasts per stomata guard cell. Further analysis by flow cytometry and chromosome counting confirmed seven out of 129 were hexadecaploids. Besides, the growth rate (GR) of PLBs in hexadecaploid plants was 10.49% (increase rate in 1 day), 1.6 times higher than that of octoploid plants and the alkaloid content of PLBs in hexadecaploid plants was 0.0794%, 1.8 times higher than that of octoploid plant. The results indicated that we have established a successful procedure for development and propagation of hexadecaploid of P. ternate with higher GR and alkaloid content.

In order to develop an efficient continuous process for producing xylitol at high yield, batch xylitol production on corn cob hemicellulosic hydrolysates carried out in a three-phase fluidized-bed bioreactor using immobilized Candida sp.ZU04 cells was investigated. It was found that two-phase aeration process was more effective than one-phase aeration process in xylitol production. In the first 24 h of aerobic phase, glucose was soon consumed and biomass in the Ca-alginate beads increased quickly at high aeration rate of 1.00 vvm. In the second fermentation phase, aeration rate was reduced to 0.30 vvm and the xylitol yield increased remarkably. The immobilized cells could be reused successfully before 6 batch cycles with average xylitol yield of 0.73 g g−1 and average volumetric productivity of 0.84 g l-1h-1 in the bioreactor for 501 h.

To the Editor, We read with great interest the two recent meta-analysis of the association between Tumor necrosis factor(TNF)-a 308G/A polymorphism and breast cancer risk [1, 2] and the two related letters [3, 4] which commented on the two articles. The meta-analysis by Fang et al. [1] included 11 studies with 10,184 breast cancer cases and 12,911 controls, and the results suggested that TNF-a308G allele might be a modestly risk factor for the development of breast cancer among Caucasian and the overall. However, the other one by Shen et al. [2] comprised 13 studies involving 10,236 cases and 13,143 controls, and the results indicated that TNF-a308A allele might slightly decrease the risk of breast cancer among Caucasian, but not among the overall. The partial difference (for the overall population) on the conclusions between two studies is due to the eligible studies they identified. As commented by Chen et al. [3] and Zhou et al. [4], the article by Fang et al. contained one study not in Hardy–Weinberg equilibrium [5] and omitted two eligible studies [6, 7]; while the article by Shen et al. [2] included one duplicate study [8]. These findings suggested that the incomprehensive search of publications would bias the results. In addition, we have three further comments on the Rebuttal Comments by Fang et al. [3] and Shen et al. [4]. First, Fang et al. [3] mentioned that the study by Kohaar et al. [6] did not supply the frequencies of GA and AA genotypes. In fact, this information was provided in the main text (see the fourth paragraph of the ‘‘Result’’ section). Second, the opinions of Shen et al. [4] suggested that PubMed searching might be enough to indentify the eligible studies. We agree that PubMed database is one of the most comprehensive sources of medicine information worldwide, while its coverage is incomplete like any database [9, 10]. Other databases, such as ISI Web of Knowledge, Embase, and even non-English databases also should be searched to avoid the introduction of selection bias and language bias [10, 11]; or at least, both Medline and hand searching should be used when the meta-analysis is performed [10]. Third, the reference number in the ‘‘References for Rebuttal letter’’ section is not in accord with that in the main text of Rebuttal letter [4].