[Development and implementation of the course entitled "Virtual Simulation Experiment of Recombinant Human Erythropoietin Manufacturing Process"].

Considering the issues present in traditional learning methods of manufacturing process for biotechnology majors, this paper presents the development and implementation process of the course entitled "Virtual Simulation Experiment of Recombinant Human Erythropoiesis Manufacturing Process". The experiment combines modern biological manufacturing technology and three-dimensional information technology, with recombinant human erythropoiesis drug serving as the focal point. This paper elaborates on the teaching concepts, objectives, contents, implementation methods, experimental procedures, interactive steps, and assessment criteria used in the experiment. Through innovative experimental scheme design, teaching methodologies, and evaluation systems, this course aims to cultivate students' analytical and problem-solving skills in the field of biopharmaceutical engineering, while also broadening students' perspective and expanding their vision.

Coexpression of Tail Fiber and Tail Protein Genes of the Cyanophage PP Using a Synthetic Genomics Approach Enhances the Salt Tolerance of Synechocystis PCC 6803.

Cyanophages are viruses that specifically infect cyanobacteria and are capable of regulating the population densities and seasonal distributions of cyanobacteria. However, few studies have investigated the interactions between cyanophages and heterologous hosts, owing to the inability of cyanophages to infect heterologous cyanobacterial hosts. Here, a truncated artificial cyanophage genome, Syn-P4-8, was designed and assembled that contained 18 genes for viral coat assembly proteins but not genes related to host infection or DNA replication. Syn-P4-8 was transferred into the heterologous host Synechocystis sp. PCC 6803 by conjugation. The growth of strain CS-02 carrying Syn-P4-8 was significantly better than that of the control strain when grown in medium containing 5% NaCl. Only two cyanophage genes, encoding the tail protein (open reading frame 25 [ORF25]) and the tail fiber protein (ORF26), were transcribed in Synechocystis PCC 6803 grown in BG11 medium supplemented with 5% NaCl. However, expression of either ORF25 or ORF26 alone could not recover this phenotype. In addition, transcriptomic analysis revealed the presence of 334 differentially expressed genes in CS-02 compared to the control strain, corresponding to 151 downregulated and 183 upregulated genes that may affect cyanobacterial salt tolerances. In this study, synthetic biology methods were used to strengthen our understanding of the interactions between cyanophage genes and heterologous hosts. IMPORTANCE We synthesized and assembled a truncated cyanophage genome called Syn-P4-8, containing 18 genes for viral coat assembly proteins, and transferred it into a nonhost strain, Synechocystis sp. PCC 6803, to investigate interactions between Syn-P4-8 and Synechocystis PCC 6803. We found that coexpression of tail fiber and tail protein genes enhanced the salt tolerance of Synechocystis PCC 6803.

Synthesis and assembly of full-length cyanophage A-4L genome.

Artificial cyanophages are considered to be an effective biological method to control harmful cyanobacterial bloom. However, no synthetic cyanophage genome has been constructed and where its obstacles are unclear. Here, we survey a stretch of 16 kb length sequence of cyanophage A-4L that is unclonable in Escherichia coli. We test 12 predicted promoters of cyanophage A-4L which were verified all active in E. coli. Next, we screen for eight ORFs that hindered the assembly of intermediate DNA fragments in E. coli and describe that seven ORFs in the 16 kb sequence could not be separately cloned in E. coli. All of unclonable ORFs in high-copy-number plasmid were successfully cloned using low-copy-number vector, suggesting that these ORFs were copy-number-dependent. We propose a clone strategy abandoned the promotor and the start codon that could be applied for unclonable ORFs. Last, we de novo synthesized and assembled the full-length genome of cyanophage A-4L. This work deepens the understanding of synthetic cyanophages studies.

Histamine bidirectionally regulates the intrinsic excitability of parvalbumin-positive neurons in the lateral globus pallidus and promotes motor behaviour.

BACKGROUND AND PURPOSE: Parvalbumin (PV)-positive neurons are a type of neuron in the lateral globus pallidus (LGP) which plays an important role in motor control. The present study investigated the effect of histamine on LGPPV neurons and motor behaviour. EXPERIMENTAL APPROACH: Histamine levels in LGP as well as its histaminergic innervation were determined through brain stimulation, microdialysis, anterograde tracing and immunostaining. Mechanisms of histamine action were detected by immunostaining, single-cell qPCR, whole-cell patch-clamp recording, optogenetic stimulation and CRISPR/Cas9 gene-editing techniques. The effect of histamine on motor behaviour was detected by animal behavioural tests. KEY RESULTS: A direct histaminergic innervation in LGP from the tuberomammillary nucleus (TMN) and a histamine-induced increase in the intrinsic excitability of LGPPV neurons were determined by pharmacological blockade or by genetic knockout of the histamine H1 receptor (H1 R)-coupled TWIK-related potassium channel-1 (TREK-1) and the small-conductance calcium-activated potassium channel (SK3), as well as by activation or overexpression of the histamine H2 receptor (H2 R)-coupled hyperpolarization-activated cyclic nucleotide-gated channel (HCN2). Histamine negatively regulated the STN → LGPGlu transmission in LGPPV neurons via the histamine H3 receptor (H3 R), whereas blockage or knockout of H3 R increased the intrinsic excitability of LGPPV neurons. CONCLUSIONS AND IMPLICATIONS: Our results indicated that the endogenous histaminergic innervation in the LGP can bidirectionally promote motor control by increasing the intrinsic excitability of LGPPV neurons through postsynaptic H1 R and H2 R, albeit its action was negatively regulated by the presynaptic H3 R, thereby suggesting possible role of histamine in motor deficits manifested in Parkinson's disease (PD).

Receptor and Ionic Mechanism of Histamine on Mouse Dorsolateral Striatal Neurons.

The dorsolateral striatum (DLS) is the critical neural substrate that plays a role in motor control and motor learning. Our past study revealed a direct histaminergic projection from the tuberomammillary nucleus (TMN) of the hypothalamus to the rat striatum. However, the afferent of histaminergic fibers in the mouse DLS, the effect of histamine on DLS neurons, and the underlying receptor and ionic mechanisms remain unclear. Here, we demonstrated a direct histaminergic innervation from the TMN in the mouse DLS, and histamine excited both the direct-pathway spiny projection neurons (d-SPNs) and the indirect-pathway spiny projection neurons (i-SPNs) of DLS via activation of postsynaptic H1R and H2R, albeit activation of presynaptic H3R suppressed neuronal activity by inhibiting glutamatergic synaptic transmission on d-SPNs and i-SPNs in DLS. Moreover, sodium-calcium exchanger 3 (NCX3), potassium-leak channels linked to H1R, and hyperpolarization-activated cyclic nucleotide-gated channel 2 (HCN2) coupled to H2R co-mediated the excitatory effect induced by histamine on d-SPNs and i-SPNs in DLS. These results demonstrated the pre- and postsynaptic receptors and their downstream multiple ionic mechanisms underlying the inhibitory and excitatory effects of histamine on d-SPNs and i-SPNs in DLS, suggesting a potential modulatory effect of the central histaminergic system on the DLS as well as its related motor control and motor learning.

The Synthesis and Assembly of a Truncated Cyanophage Genome and Its Expression in a Heterogenous Host.

Cyanophages play an important role in regulating the dynamics of cyanobacteria communities in the hydrosphere, representing a promising biological control strategy for cyanobacterial blooms. Nevertheless, most cyanophages are host-specific, making it difficult to control blooming cyanobacteria via single or multiple cyanophages. In order to address the issue, we explore the interaction between cyanophages and their heterologous hosts, with the aim of revealing the principles of designing and constructing an artificial cyanophage genome towards multiple cyanobacterial hosts. In the present study, we use synthetic biological approaches to assess the impact of introducing a fragment of cyanophage genome into a heterologous cyanobacterium under a variety of environmental conditions. Based on a natural cyanophage A-4L genome (41,750 bp), a truncated cyanophage genome Syn-A-4-8 is synthesized and assembled in Saccharomyces cerevisiae. We found that a 351-15,930 bp area of the A-4L genome has a fragment that is lethal to Escherichia coli during the process of attempting to assemble the full-length A-4L genome. Syn-A-4-8 was successfully introduced into E. coli and then transferred into the model cyanobacterium Synechococcus elongatus PCC 7942 (Syn7942) via conjugation. Although no significant phenotypes of Syn7942 carrying Syn-A-4-8 (LS-02) could be observed under normal conditions, its growth exhibited a prolonged lag phase compared to that of the control strain under 290-millimolar NaCl stress. Finally, the mechanisms of altered salt tolerance in LS-02 were revealed through comparative transcriptomics, and ORF25 and ORF26 on Syn-A-4-8 turned out to be the key genes causing the phenotype. Our research represents an important attempt in designing artificial cyanophages towards multiple hosts, and offers new future insights into the control of cyanobacterial blooms.

Development and validation of a liquid chromeatography-tandem mass spectrometry method for simultaneous quantification of medium- and long-chain saturated fatty acids in hamster plasma samples.

RATIONALE: Saturated fatty acids (SFAs) are associated with many diseases in humans. Developing a reliable analytical method to analyze SFAs in plasma is essential to understand their biological activities. An ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC/MS/MS) method has been developed for the quantification of medium- and long-chain SFAs (M/LCSFAs) in hamster plasma. METHODS: We compared three methods (DOLE, Folch and MTBE) for extracting M/LCSFAs from plasma. The M/LCSFA derivatives were separated using a C18 column. The method was validated and applied to analyze M/LCSFA concentrations in normal-fat diet (NFD) and high-fat diet (HFD) hamster plasma. RESULTS: Among the three extraction methods, the DOLE method had the highest extraction recovery and was simple to operate with a short incubation time. All of the calibration curves exhibited good linear relationships (r ≥ 0.9958). The results for selectivity, accuracy, precision, matrix effects and recovery were all within the acceptance criteria. In HFD hamster plasma, the concentration of M/LCSFAs with even-carbon chain length was significantly increased. CONCLUSIONS: A simple, robust and reproducible method for the simultaneous quantification of M/LCSFAs by UHPLC/MS/MS was developed and validated. The method gave successfully quantification of M/LCSFAs in plasma samples from NFD and HFD hamsters.

Diversity-oriented synthesis of marine sponge derived hyrtioreticulins and their anti-inflammatory activities.

Diversity-oriented synthesis is aimed to increase the chemical diversity of target natural products for extensive biological activity evaluation. Indole ring is an important functional group in a large number of drugs and other biologically active agents, and indole-containing natural products have been frequently isolated from marine sources in recent years. In this paper, a series of indole-containing marine natural hyrtioreticulin derivatives, including 19 new ones, were designed, synthesized through a key Pictet-Spengler reaction, and evaluated for their inflammation related activity. Compound 13b displayed the most promising activity by inhibiting TNF-α cytokine release with an inhibitory rate of 92% at a concentration of 20 µmol·L-1. A preliminary structure-activity relationship analysis was also discussed. This research may throw light on the discovery of marine indole alkaloid derived anti-inflammatory drug leads.

A simultaneous identification and quantification strategy for determination of sulfhydryl-containing metabolites in normal- and high-fat diet hamsters using stable isotope labeling combined with LC-MS.

Sulfur-containing metabolites are related to several physiologic disorders and metabolic diseases. In this study, a simultaneous identification and quantification strategy in one batch for determination of sulfhydryl-containing metabolites was developed using stable isotope labeling combined with liquid chromatography-tandem mass spectrometry (SIL-LC-MS). In the proposed method, a pair of isotope labeling reagents, D0/D5-N-ethylmaleimide (D0/D5-NEM), was used to derivatize sulfhydryl-containing metabolites in blood and plasma of normal- and high-fat-diet (NFD and HFD) hamsters for reduced (-SH) and total (-SH, -S-S-, S-glutathionylated proteins) analysis. Quality control (QC) samples and test samples were prepared for LC-MS analysis. First, both QC samples and stable isotope labeled internal standards were used to monitor the status of the instrument and ensure the reliability of the analysis. Subsequently, an inhouse database containing 45 sulfhydryl-containing metabolites was established by MS1 based on QC samples. Then, qualitatively differential sulfhydryl-containing metabolites were found by MS2 between the NFD and HFD hamsters of the test samples, including 3 in reduced and 8 in total analysis of blood samples, and 2 in reduced and 2 in total analysis of plasma samples. Next, in quantitative analysis, satisfied linearities for 6 sulfhydryl-containing metabolites were obtained with the correlation coefficient (R2) > 0.99 and absolute quantification was carried out. The results showed that glutathione and cysteine have different concentrations in blood and plasma of hamsters. Finally, the correlation of sulfhydryl-containing metabolites with blood lipid and oxidative stress levels was determined, which provided insight into the hyperlipidemia-related oxidative stress. Taken together, the developed method of simultaneous identification with the inhouse database and MS2 and quantification with standards in one batch provides a promising strategy for the analysis of sulfhydryl-containing metabolites in biological samples, which may promote the in-depth investigation on sulfhydryl-containing metabolites and related diseases.

Qualitatively and quantitatively investigating the metabolism of 20(S)-protopanaxadiol-type ginsenosides by gut microbiota of different species.

Ginsenosides Rb1, Rb2, Rb3 and Rc, four major protopanaxadiol (PPD)-type ginsenosides, can be metabolized by gut microbiota. The composition of gut microbiota varies in different species. Existing publications have reported the metabolite fates of ginsenosides by gut microbiota from single species. However, their microbiota-related metabolic species differences have not been evaluated yet. In current study, in vitro anaerobic incubations of PPD-type ginsenosides with gut microbiota from humans, rabbits and rats were conducted. The metabolites of each ginsenoside were then identified by LC-MS. A total of 15 metabolites from the four ginsenosides were identified. The major metabolic pathways were stepwise removals of the C-20 and C-3 sugar moieties to obtain aglycone PPD. The results showed that the hydrolysis rate of C-20 terminal ß-D-glucopyranosyl was significantly higher than those of α-L-arabinopyranosyl, ß-D-xylopyranosyl and α-L-arabinofuranosyl in different species. The activity of ß-glucosidase, the metabolic rates of parent compounds and the formation rates of their metabolites were significantly higher in gut microbiota from rabbits than from humans and rats. Our research draws researchers' attention to the species differences of microbiota-related drug metabolism.

Two novel spirobifluorene-based two-photon fluorescent probes for the detection of hydrazine in solution and living cells.

Two spirobifluorene-based two-photon fluorescent probes for the detection of hydrazine, namely SPF-MN and SPF- PA, have been designed and synthesized. Along with the addition of hydrazine to a solution of SPF-MN, both a colorimetric change from yellow to colorless and a fluorescence change from yellow to blue (Under 365 nm UV light) can be observed by ''naked-eye''. Probe SPF- PA displayed response toward hydrazine with fluorescence enhancement. The detection limits are 6.9 µM for SPF- PA and 0.29 µM for SPF-MN, respectively. Moreover, SPF-MN and SPF- PA can be used as two-photon fluorescent probes for hydrazine with large two-photon absorption cross-sections and used for the imaging of hydrazine in living cells. Specially, SPF-PA can located at the surface of the cells, and it is the first fluorescent probe which possesses the capability of sensing intercellular hydrazine. Besides, SPF-MN is the first colorimetric two-photon fluorescent probe for meeting the criteria of both hydrazine bioimaging and visual detection of hydrazine in solution.

The biodegradation of cefuroxime, cefotaxime and cefpirome by the synthetic consortium with probiotic Bacillus clausii and investigation of their potential biodegradation pathways.

Cephalosporin residues in the environment are a great concern, but bioremediation options do exist. Bacillus clausii T reached a removal rate of 100% within 8 h when challenged with a mixture of cefuroxime (CFX), cefotaxime (CTX), and cefpirome (CPR). The co-culture of B. clausii T and B. clausii O/C displayed a higher removal efficiency for the mixture of CFX, CTX and CPR than a pure culture of B. clausii O/C. B. clausii T alleviated the biotoxicity of CFX and CPR. What's more, the biotoxicity of for CFX and CPR transformation products released by the co-culture of B. clausii T and B. clausii O/C was lower than that in pure cultures. Real-time PCR was applied to detect the changes in the expression levels of the relevant antibiotic-resistance genes of B. clausii T during CFX and CPR degradation. The results indicated that CFX and CPR enhanced the expression of the ß-lactamase gene bcl1. Hydrolysis, deacetylation and decarboxylation are likely the major mechanisms of CTX biodegradation by B. clausii. These results demonstrate that B. clausii T is a promising strain for the bioremediation of environmental contamination by CFX, CTX, and CPR.

The application of SVR model in the improvement of QbD: a case study of the extraction of podophyllotoxin.

In order to make a further optimization of process design via increasing the stability of design space, we brought in the model of Support Vector Regression (SVR). In this work, the extraction of podophyllotoxin was researched as a case study based on Quality by Design (QbD). We compared the fitting effect of SVR and the most used quadratic polynomial model (QPM) in QbD, and an analysis was made between the two design spaces obtained by SVR and QPM. As a result, the SVR stayed ahead of QPM in prediction accuracy, the stability of model and the generalization ability. The introduction of SVR into QbD made the extraction process of podophyllotoxin well designed and easier to control. The better fitting effect of SVR improved the application effect of QbD and the universal applicability of SVR, especially for non-linear, complicated and weak-regularity problems, widened the application field of QbD.

Convergent engineering of syntrophic Escherichia coli coculture for efficient production of glycosides.

Synthetic microbial coculture to express heterologous biosynthetic pathway for de novo production of medicinal ingredients is an emerging strategy for metabolic engineering and synthetic biology. Here, taking efficient production of salidroside as an example of glycosides, we design and construct a syntrophic Escherichia coli-E. coli coculture composed of the aglycone (AG) strain and the glycoside (GD) strain, which convergently accommodate biosynthetic pathways of tyrosol and salidroside, respectively. To accomplish this the phenylalanine-deficient AG strain was engineered to utilize xylose preferentially and to overproduce precursor tyrosol, while the tyrosine-deficient GD strain was constructed to consume glucose exclusively and to enhance another precursor UDP-glucose availability for synthesis of salidroside. The AG and GD strains in the synthetic consortium are obligatory cooperators through crossfeeding of tyrosine and phenylalanine and compatible in glucose and xylose mixture. Through balancing the metabolic pathway strength, we show that the syntrophic coculture was robust and stable, and produced 6.03 g/L of salidroside. It was the de novo production of salidroside for the first time in E. coli coculture system, which would be applicable for production of other important glycosides and natural products.

Antitumor component recognition from the Aconiti Lateralis Radix Praeparata and Glycyrrhizae Radix et Rhizoma herb pair extract by chemometrics and mean impact value.

The purpose of this research is to recognize the active antitumor components from the mixed pair extract of Aconiti Lateralis Radix Praeparata (Fuzi in Chinese) and Glycyrrhizae Radix et Rhizoma (Gancao in Chinese) using chemometrics and mean impact value (MIV) methods. Firstly, 30 common components of 31 different samples were analyzed quantitatively and qualitatively by HPLC-UV and UPLC-Q-TOF tandem mass spectrometry, respectively. Meanwhile, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays were used to test the inhibition activities of the 31 different samples against HeLa cells. Then a back propagation (BP) neural network, support vector regression (SVR), and two optimization algorithms - genetic algorithm (GA) and particle swarm optimization (PSO) - were applied to construct composition-activity relationship (CAR) models for the Fuzi-Gancao extract. Based on the optimal CAR model, the MIV was introduced to evaluate the contribution of each individual component to the anticancer efficacy of the extract. Results indicated that the SVR-PSO model best depicted the complex relationship between the chemical composition and the inhibition effect of a Fuzi-Gancao extract. The 30 common components were ranked by their absolute MIVs, and the top 8, which corresponded to peaks 17, 25, 22, 13, 23, 28, 5, and 7 in the chromatogram, were tentatively deemed to be the main antitumor components. The integrated strategy shows a novel and efficient approach to understanding the potential contributions of components from complicated herbal medicines, and the identified results suggest certain directions for screening and research into new antitumor drugs.

[Extraction Optimization of Rhizome of Curcuma longa by Response Surface Methodology and Support Vector Regression].

OBJECTIVE: To optimize the optimal microwave-assisted extraction method of curcuminoids from Curcuma longa. METHODS: On the base of single factor experiment, the ethanol concentration, the ratio of liquid to solid and the microwave time were selected for further optimization. Support Vector Regression (SVR) and Central Composite Design-Response Surface Methodology (CCD) algorithm were utilized to design and establish models respectively, while Particle Swarm Optimization (PSO) was introduced to optimize the parameters of SVR models and to search optimal points of models. The evaluation indicator, the sum of curcumin, demethoxycurcumin and bisdemethoxycurcumin by HPLC, were used. RESULTS: The optimal parameters of microwave-assisted extraction were as follows: ethanol concentration of 69%, ratio of liquid to solid of 21 : 1, microwave time of 55 s. On those conditions, the sum of three curcuminoids was 28.97 mg/g (per gram of rhizomes powder). CONCLUSION: Both the CCD model and the SVR model were credible, for they have predicted the similar process condition and the deviation of yield were less than 1.2%.

[Chromatographic Fingerprinting Study of Zhenyuan Granules Dry Extract by HPLC-DAD and HPLC-MS/MS].

OBJECTIVE: To establish a novel, accurate and valid fingerprint method of Zhenyuan granules dry extract by using HPLC-DAD method, to study herbs belonging of fingerprint peaks and to identify some of the chromatographic peaks by HPLC-MS/MS analysis, for providing the basis for scientific evaluation of the quality. METHODS: The sample solutions were analyzed by an Agilent SB C18 (250 mm x 4.6 mm, 5 µm) column, and gradiently eluted with acetonitrile (containing 0.1% formic acid) and aqueous phase (containing 0.1% formic acid) as the mobile phase. The flow rates were 1.2 mL/min (0~70 min) and 0.8 mL/min (70~150 min); the column temperature was 30 °C; and the detection wavelength was 254 nm. RESULTS: 40 peaks were selected as fingerprint peaks under the optimal chromatographic condition, and the similarity coefficients of 10 batches of Zhenyuan granules dry extract were all greater than 0.98. 27 peaks were tentatively identified with reference to literature data based on their mass spectrometry. CONCLUSION: The chromatographic fingerprint of Zhenyuan granules is proved to be a reliable method for comprehensive quality control and assessment.

Feature selection for the identification of antitumor compounds in the alcohol total extracts of Curcuma longa.

Antitumor activity has been reported for turmeric, the dried rhizome of Curcuma longa. This study proposes a new feature selection method for the identification of the antitumor compounds in turmeric total extracts. The chemical composition of turmeric total extracts was analyzed by gas chromatography-mass spectrometry (21 ingredients) and high-performance liquid chromatography-mass spectrometry (22 ingredients), and their cytotoxicity was detected through an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay against HeLa cells. A support vector machine for regression and a generalized regression neural network were used to research the composition-activity relationship and were later combined with the mean impact value to identify the antitumor compounds. The results showed that six volatile constituents (three terpenes and three ketones) and seven nonvolatile constituents (five curcuminoids and two unknown ingredients) with high absolute mean impact values exhibited a significant correlation with the cytotoxicity against HeLa cells. With the exception of the two unknown ingredients, the identified 11 constituents have been reported to exhibit cytotoxicity. This finding indicates that the feature selection method may be a supplementary tool for the identification of active compounds from herbs.

Chronic atrophic gastritis in association with hair mercury level.

The objective of this study was to explore hair mercury level in association with chronic atrophic gastritis, a precancerous stage of gastric cancer (GC), and thus provide a brand new angle of view on the timely intervention of precancerous stage of GC. We recruited 149 healthy volunteers as controls and 152 patients suffering from chronic gastritis as cases. The controls denied upper gastrointestinal discomforts, and the cases were diagnosed as chronic superficial gastritis (n=68) or chronic atrophic gastritis (n=84). We utilized Mercury Automated Analyzer (NIC MA-3000) to detect hair mercury level of both healthy controls and cases of chronic gastritis. The statistic of measurement data was expressed as mean ± standard deviation, which was analyzed using Levene variance equality test and t test. Pearson correlation analysis was employed to determine associated factors affecting hair mercury levels, and multiple stepwise regression analysis was performed to deduce regression equations. Statistical significance is considered if p value is less than 0.05. The overall hair mercury level was 0.908949 ± 0.8844490 ng/g [mean ± standard deviation (SD)] in gastritis cases and 0.460198 ± 0.2712187 ng/g (mean±SD) in healthy controls; the former level was significantly higher than the latter one (p=0.000<0.01). The hair mercury level in chronic atrophic gastritis subgroup was 1.155220 ± 0.9470246 ng/g (mean ± SD) and that in chronic superficial gastritis subgroup was 0.604732 ± 0.6942509 ng/g (mean ± SD); the former level was significantly higher than the latter level (p<0.01). The hair mercury level in chronic superficial gastritis cases was significantly higher than that in healthy controls (p<0.05). The hair mercury level in chronic atrophic gastritis cases was significantly higher than that in healthy controls (p<0.01). Stratified analysis indicated that the hair mercury level in healthy controls with eating seafood was significantly higher than that in healthy controls without eating seafood (p<0.01) and that the hair mercury level in chronic atrophic gastritis cases was significantly higher than that in chronic superficial gastritis cases (p<0.01). Pearson correlation analysis indicated that eating seafood was most correlated with hair mercury level and positively correlated in the healthy controls and that the severity of gastritis was most correlated with hair mercury level and positively correlated in the gastritis cases. Multiple stepwise regression analysis indicated that the regression equation of hair mercury level in controls could be expressed as 0.262 multiplied the value of eating seafood plus 0.434, the model that was statistically significant (p<0.01). Multiple stepwise regression analysis also indicated that the regression equation of hair mercury level in gastritis cases could be expressed as 0.305 multiplied the severity of gastritis, the model that was also statistically significant (p<0.01). The graphs of regression standardized residual for both controls and cases conformed to normal distribution. The main positively correlated factor affecting the hair mercury level is eating seafood in healthy people whereas the predominant positively correlated factor affecting the hair mercury level is the severity of gastritis in chronic gastritis patients. That is to say, the severity of chronic gastritis is positively correlated with the level of hair mercury. The incessantly increased level of hair mercury possibly reflects the development of gastritis from normal stomach to superficial gastritis and to atrophic gastritis. The detection of hair mercury is potentially a means to predict the severity of chronic gastritis and possibly to insinuate the environmental mercury threat to human health in terms of gastritis or even carcinogenesis.

HPLC-MS and GC-MS analyses combined with orthogonal partial least squares to identify cytotoxic constituents from turmeric (Curcuma longa L.).

We investigated the fingerprints of 48 batches of turmeric total extracts (TTE) by HPLC-MS-MS and GC-MS analyses and 43 characteristic peaks (22 constituents from HPLC-MS-MS; 21 from GC-MS) were analyzed qualitatively and quantitatively. An MTT {3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide} assay was implemented to measure the cytotoxicity of the TTE against HeLa cells. Then we utilized orthogonal partial least squares analysis, which correlated the chemical composition of the TTE to its cytotoxic activity, to identify potential cytotoxic constituents from turmeric. The result showed that 19 constituents contributed significantly to the cytotoxicity. The obtained result was verified by canonical correlation analysis. Comparison with previous reports also indicated some interaction between the curcuminoids and sesquiterpenoids in turmeric.