Association Between Serum Glycated Hemoglobin Levels at Early Gestation and the Risk of Subsequent Pregnancy Loss in Pregnant Women Without Diabetes Mellitus: Prospective Cohort Study.

BACKGROUND: As a severe morbidity during pregnancy, the etiology of spontaneous pregnancy loss (SPL) remains largely unknown. Serum glycated hemoglobin (HbA1c) level is an established predictor of SPL risk among women with diabetes, but little is known about whether such an association exists among pregnant women without diabetes when glycemic levels are within the normal range. OBJECTIVE: This study aimed to quantify the association between maternal HbA1c levels in early pregnancy and subsequent SPL risk in a cohort of pregnant women without diabetes. METHODS: This prospective cohort study involved 10,773 pregnant women without diabetes enrolled at their first antenatal care visit at a hospital's early pregnancy clinic from March 2016 to December 2018 in Shanghai, China. HbA1c and fasting blood glucose (FBG) levels were examined at enrollment. Participants with diabetes before or pregnancy or those diagnosed with gestational diabetes were excluded. Diagnosis of SPL, defined as fetal death occurring before 28 gestational weeks, was derived from medical records and confirmed via telephone interviews. We used generalized linear models to quantify the associations of continuous and dichotomized maternal HbA1c levels with SPL risk and reported crude and adjusted risk ratios (RRs) and 95% CIs. A restricted cubic spline (RCS) regression model was used to assess the potential nonlinear dose-response relationship. Adjusted covariates included maternal age, education level, preconception BMI, gestational weeks, gravidity, history of adverse pregnancy outcomes, family history of diabetes, folic acid supplementation, and smoking and drinking during the periconception period. RESULTS: In total, 273 (2.5%) SPL cases occurred. Every 0.5% increase in HbA1c levels was linearly associated with a 23% increase in SPL risk (adjusted RR [aRR] 1.23; 95% CI 1.01-1.50). The RCS model revealed that this association was linear (P=.77 for the nonlinearity test). Analyses based on dichotomized HbA1c levels showed a significantly increased risk of SPL when HbA1c levels were ≥5.9% (aRR 1.67; 95% CI 0.67-3.67), and the significance threshold was ≥5.6% (aRR 1.60; 95% CI 1.01-2.54). Sensitivity analyses showed similar results when including the participants with missing SPL records or HbA1c data. Linear associations of HbA1c levels remained significant even in the subgroups without overweight, alcohol consumption, and a family history of diabetes and adverse pregnancy outcomes. Every 1 mmol/L increment in maternal FBG levels was associated with a >2-fold higher risk of SPL (aRR 2.12; 95% CI 1.61-2.80; P<.001). CONCLUSIONS: Higher HbA1c levels in early pregnant women without diabetes are associated with an increased SPL risk in a dose-response manner. Pregnant women with an HbA1c level above 5.6% at early gestation need attention for its potentially increased risk for SPL. Our findings support the need to monitor HbA1c levels to identify individuals at high risk of subsequent SPL in the general population of pregnant women. TRIAL REGISTRATION: ClinicalTrials.gov NCT02737644; https://clinicaltrials.gov/study/NCT02737644.

Impact of Nutrition Counseling in Head and Neck Cancer Sufferers Undergoing Antineoplastic Therapy: A Randomized Controlled Pilot Study.

Head and neck cancer (HNC) sufferers usually encounter arduous nutritional problems when they are receiving antineoplastic therapy. Consequently, the presence of anxiety and depression is commonly observed in this population. This study aimed to explore the physical and psychological influence of nutritional counseling in patients with HNC. Patients receiving concurrent chemo-radiotherapy were randomly assigned to the nutritional counseling group (n = 32, 52.45%) and the control group (n = 29, 47.54%) according to their treatment patterns. In the nutritional counseling group, registered dietitians provided face-to-face counseling during the antineoplastic treatment course at least every two weeks. Nutrient intake amount, relevant nutritional indexes, quality of life, and the degree of anxiety and depression were compared between the two groups. We observed a decrease in the calorie and protein intake amount in both groups, while the decrease in the control group is even worse. The weight loss is more obvious in the control group. The HADS scores in the intervention group were significantly lower than that in the control group (p < 0.05). The Karnofsky Performance Status (KPS) scores in the intervention group were significantly higher than that in the control group (p < 0.05). The level of serum total protein, serum albumin, transferrin, and the thickness of the triceps skin fold decreased less in the intervention group (p < 0.05). Our findings suggest that nutrition counseling is essential for the maintenance of calorie and protein intake in HNC suffers, which contributes to an improvement in the physical and psychological states. The impacts observed in this pilot study warrant further exploration in a larger prospective trial.

Periconception Red Blood Cell Folate and Offspring Congenital Heart Disease : Nested Case-Control and Mendelian Randomization Studies.

BACKGROUND: Periconception folic acid supplementation has been suggested to protect against congenital heart disease (CHD), but the association between maternal red blood cell (RBC) folate, the gold-standard biomarker of folate exposure, and subsequent offspring CHD risk is lacking. OBJECTIVE: To quantify the association between periconception maternal RBC folate and offspring CHD risk. DESIGN: Prospective, nested, case-control study and 1-sample Mendelian randomization. (ClinicalTrials.gov: NCT02737644). SETTING: 29 maternity institutions in 12 districts of Greater Shanghai, China. PARTICIPANTS: All 197 mothers of offspring with CHD and 788 individually matched mothers of unaffected offspring from the SPCC (Shanghai Preconception Cohort). MEASUREMENTS: Maternal RBC folate was measured before or at early pregnancy. Odds ratios [ORs] were estimated using conditional logistic regression after adjustment for covariates. Mendelian randomization was done using the methylenetetrahydrofolate reductase (MTHFR) C677T as the genetic instrument. RESULTS: Case patients had lower median maternal RBC folate concentrations than control participants (714 nmol/L [interquartile range, 482 to 1008 nmol/L] vs. 788 nmol/L [557 to 1094 nmol/L]). Maternal RBC folate concentrations were inversely associated with offspring CHD (adjusted OR per 100 nmol/L, 0.93 [95% CI, 0.89 to 0.99]). The adjusted OR for mothers with periconception RBC folate of 906 nmol/L or more (vs. <906 nmol/L) was 0.61 (CI, 0.40 to 0.93). Mendelian randomization showed that each 100-nmol increase in maternal RBC folate concentrations was significantly associated with reduced offspring CHD risk (OR, 0.75 [CI, 0.61 to 0.92]). LIMITATION: Potential confounding due to unmeasured covariates in the nested case-control study. CONCLUSION: Higher maternal RBC folate is associated with reduced offspring CHD risk. For primary CHD prevention, higher target RBC folate levels than currently recommended for neural tube defect prevention may be needed and warrant further study. PRIMARY FUNDING SOURCE: National Key Research and Development Program of China, National Natural Science Foundation of China, China Postdoctoral Science Foundation, and Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences.

RBC Folate and Serum Folate, Vitamin B-12, and Homocysteine in Chinese Couples Prepregnancy in the Shanghai Preconception Cohort.

BACKGROUND: The protective effects of maternal folate on neural tube defects are well-established. Emerging evidence has shown paternal folate also is related to pregnancy outcome and offspring health. OBJECTIVES: This study aimed to assess the status of red blood cell (RBC) folate and serum folate, vitamin B-12, and homocysteine (Hcy) and their associated factors in a cohort of pregnancy-preparing couples. METHODS: This was a cross-sectional study involving 14,178 participants from the extension of the Shanghai Preconception Cohort conducted in 2018-2021. Circulating biomarker concentrations were measured, and the prevalence of abnormal status was reported. Linear and logistic regression analyses were conducted to examine associations of demographic factors (age, education, and income), lifestyle factors (smoking, drinking, and folic acid supplement use), and BMI with concentrations of the folate-related biomarkers, abnormal status of folate (deficiency and insufficiency) and vitamin B-12 (deficiency and marginal deficiency), and hyperhomocysteinemia. RESULTS: The geometric mean (95% CI) concentrations of RBC folate, serum folate, vitamin B-12, and Hcy were 490 nmol/L (485, 496 nmol/L), 20.1 nmol/L (19.8, 20.3 nmol/L), 353 pmol/L (350, 357 pmol/L), and 7.54 µmol/L (7.48, 7.60 µmol/L) in females, respectively, and 405 nmol/L (401, 409 nmol/L), 13.5 nmol/L (13.4, 13.7 nmol/L), 277 pmol/L (274, 279 pmol/L), and 12.0 µmol/L (11.9, 12.2 µmol/L) in males, respectively. Prevalence of abnormal status was higher in males than females for the 4 folate-related biomarkers: RBC folate deficiency (<340 nmol/L, 32.2% compared with 18.9%), serum folate deficiency (<10.0 nmol/L, 26.5% compared with 7.3%), RBC folate insufficiency (<906 nmol/L, 96.6% compared with 90.1%), serum folate insufficiency (<15.9 nmol/L, 65.5% compared with 31.4%), vitamin B-12 marginal deficiency (148-221 pmol/L, 21.4% compared with 8.8%), and hyperhomocysteinemia (>15.0 µmol/L, 22.1% compared with 2.5%). CONCLUSIONS: Most pregnancy-preparing couples failed to achieve the optimal RBC folate status (>906 nmol/L) as recommended by the WHO. These findings call for attention to the insufficiency status of folate and promising strategies to improve the folate status of the pregnancy-preparing population not exposed to folic acid fortification.

Separation and enrichment of sibiskoside from Sibiraea angustat with magnetic surface dummy template molecularly imprinted polymers.

In this work, a novel strategy was developed for separation and enrichment of sibiskoside by dummy molecular imprinting technology and magnetic separation technology. The structural analogue geniposide was selected as the dummy template, using 4-vinylpyridine as the functional monomer, ethylene glycol dimethacrylate as the cross-linking agent, and acetonitrile as the porogen. The molecularly imprinted layer was formed on the surface of the magnetic carrier to prepare dummy template molecularly imprinted polymers (DMIPs) with a core-shell structure. The DMIPs were characterized by scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA) and Vibration sample magnetometer (VSM). The results of adsorption kinetics experiments and isothermal adsorption experiments showed that DMIPs can reach adsorption equilibrium in a short period of time and the maximum adsorption capacity can reach 14.67 mg/g. The imprinting factor was 2.08. Compared with the andrographolide, polydatin, arbutin, caffeic acid, neohesperidin dihydrochalcone and quercetin, DMIPs have good adsorption capacity for the sibiskoside. And the reusability was better. After the adsorption of DMIPs, the purity of sibiskoside in the crude extracts from Sibiraea angustata increased to 78%. It provided a basis for the further development and utilization of Sibiraea angustata as well as the separation and enrichment of monoterpenes.

Six new anthraquinone glycosides from Lasianthus acuminatissimus Merr.

Six new anthraquinones named lasianthuoside F (1), G (2), H (3), I (4), J (5), K (6) were isolated from an acetone extract of the root of Lasianthus acuminatissimus. Their structures were elucidated by physical and chemical evidence and spectral analysis.

Association of Maternal Folate and Vitamin B12 in Early Pregnancy With Gestational Diabetes Mellitus: A Prospective Cohort Study.

OBJECTIVE: To investigate the association of folate and vitamin B12 in early pregnancy with gestational diabetes mellitus (GDM) risk. RESEARCH DESIGN AND METHODS: The data of this study were from a subcohort within the Shanghai Preconception Cohort Study. We included pregnancies with red blood cell (RBC) folate and vitamin B12 measurements at recruitment (between 9 and 13 gestational weeks) and those with three samples available for glucose measurements under an oral glucose tolerance test. GDM was diagnosed between 24 and 28 weeks' gestation. Odds ratio (OR) and 95% CI of having GDM was used to quantify the association. RESULTS: A total of 1,058 pregnant women were included, and GDM occurred in 180 (17.01%). RBC folate and vitamin B12 were significantly higher in pregnancies with GDM than those without GDM (P values were 0.045 and 0.002, respectively) and positively correlated with 1-h and 2-h serum glucose. Daily folic acid supplementation in early pregnancy increases the risk of GDM; OR (95% CI) was 1.73 (1.19-2.53) (P = 0.004). Compared with RBC folate <400 ng/mL, pregnancies with RBC folate ≥600 ng/mL were associated with ∼1.60-fold higher odds of GDM; the adjusted OR (95% CI) was 1.58 (1.03-2.41) (P = 0.033). A significant trend of risk effect on GDM risk across categories of RBC folate was observed (P trend = 0.021). Vitamin B12 was significantly associated with GDM risk (OR 1.14 per 100 pg/mL; P = 0.002). No significant association of serum folate and percentile ratio of RBC folate/vitamin B12 with GDM was observed. CONCLUSIONS: Higher maternal RBC folate and vitamin B12 levels in early pregnancy are significantly associated with GDM risk, while the balance of folate/vitamin B12 is not significantly associated with GDM.

Effectiveness of community-based folate-oriented tertiary interventions on incidence of fetus and birth defects: a protocol for a single-blind cluster randomized controlled trial.

BACKGROUND: Birth defects are the main cause of fetal death, infant mortality and morbidity worldwide. However, the etiology of birth defects remains largely unknown. Maternal folate status during periconception plays an important role in organogenesis and folic acid supplement reduces the risk of neural tube defects, congenital heart diseases, and several other birth defects. This trial seeks to evaluate the effectiveness of folate-oriented tertiary interventions during periconception on the incidence of fetus and birth defects. METHODS: This is a single-blind, two-arm cluster randomized controlled trial in Shanghai, China. Eligible women from 22 clusters are recruited at pre-pregnancy physical examinations clinical settings. Compared to the routine perinatal care group (control arm), folate-oriented tertiary interventions will be provided to the intervention arm. The core interventions consist of assessments of folate status and metabolism, folate intake guidance, and re-evaluation of folate status to ensure red blood cell folate level above 400 ng/ml (906 nmol/L) before pregnancy. Screening and consulting of fetus and birth defects, and treatments of birth defects during pregnancy and afterward will be provided to both arms. The primary outcome is a composite incidence of fetus defects, stillbirth, and neonatal birth defects identified from the confirmation of pregnancy to 28 days after birth. Secondary outcomes include maternal and offspring adverse complications and cost-effectiveness of folate-oriented tertiary interventions. This protocol adheres to the SPIRIT Checklist. DISCUSSION: To achieve the recommended folate status before or during pregnancy is still a challenge worldwide. This community-based cluster-randomized controlled intervention trial will evaluate the effectiveness of a package of interventions aiming at achieving recommended maternal folate status covering pre- and during pregnancy in reducing fetus and birth defects. Our study has the potential to improve the community-based practice of reducing modifiable risk factors of disease and improving primary prevention of the defects in China. The procedures would formulate the policy on folic acid supplementation during periconception against birth defects in primary care settings. TRIAL REGISTRATION: Clinical Trial Registry, NCT03725878 . Prospectively registered on 31 October 2018.

Maternal folate levels during pregnancy and children's neuropsychological development at 2 years of age.

OBJECTIVE: To explore the relationship between maternal folate levels during pregnancy and children's neuropsychological development at 2 years of age. METHODS: In the birth cohort MKFOAD, maternal serum folate concentrations at 12-14, 22-26, and 34-36 weeks of gestation were measured, as well as red blood cell (RBC) folate at 12-14 weeks. Neurodevelopment of 2-year-old children was assessed by Gesell Development Scale (GDS), which contained subscales of gross motor, fine motor, language, adaptive behavior, and social behavior. Linear regression models were applied to investigate the association of maternal folate levels with children's developmental quotients (DQs). RESULTS: One hundred and eighty singleton children participated the GDS assessment, of whom 97 (53.9%) were boys. Median RBC folate concentration was 1002.8 (IQR = 577.6) nmol L-1 in early pregnancy and median serum folate concentrations were, respectively, 33.9 (IQR = 9.2) nmol L-1, 26.3 (IQR = 14.3) nmol L-1, and 26.7 (IQR = 18.9) nmol L-1. Maternal serum folate concentration in late pregnancy was significantly associated with children's language development, where language DQ increases by 3.1 (95% CI 0.6, 5.5) for every 10 nmol L-1 increment of serum folate concentration. And maternal serum folate in early pregnancy was significantly associated with children's fine motor development, with 2.0 (95% CI 0.1, 4.0) DQ decrease for 10 nmol L-1 increase of serum folate. CONCLUSIONS: Maternal serum folate in late pregnancy was significantly associated with children's language development at age 2, which supports the importance of remaining folic acid supplementation across the entire gestation. However, maternal serum folate in early pregnancy was also inversely associated with children's fine motor development.

Antioxidant and anti-inflammatory activities of an anti-diabetic polysaccharide extracted from Gynostemma pentaphyllum herb.

This work aimed to determine the antioxidant and anti-inflammatory effects of an anti-diabetic polysaccharide (GPP) extracted from Gynostemma pentaphyllum herb on diabetic mice. In vitro antioxidant assays showed that the DPPH radical, hydroxyl radical, superoxide anion, and ABTS radical scavenging activities of GPP increased in a dose-dependent manner. Animal experiments demonstrated that GPP could efficiently reduce the body weight and fasting blood sugar of diabetic mice and decrease the blood parameters of ALP, ALT, AST, and BUN. Furthermore, antioxidant and anti-inflammatory results indicated that GPP might exert its hypoglycemic effect by enhancing the SOD, CAT, and GSH-Px activities, decreasing the MDA activity, improving the levels of the anti-inflammatory factors IL-4 and IL-10, and decreasing the levels of the pro-inflammatory factors TNF-α and IL-6. This work provided a good basis for further revealing the prevention and treatment mechanisms of diabetes.

Shanghai Preconception Cohort (SPCC) for the association of periconceptional parental key nutritional factors with health outcomes of children with congenital heart disease: a cohort profile.

PURPOSE: The Shanghai Preconception Cohort (SPCC) was initially established to investigate the associations of parental periconceptional nutritional factors with congenital heart disease (CHD) but has further analysed child growth and development and paediatric diseases. PARTICIPANTS: Preparing-for-pregnancy couples who presented at Shanghai preconception examination clinics and early-pregnancy women before 14 gestational weeks were enrolled to comprise the periconceptional baseline study population. General characteristics, routine clinical data and consumption of diet supplements, such as folic acid and multivitamins, were collected. Blood samples were obtained at preconception and early, middle and late gestations using standard procedures. Multiple nutritional factors, including folate, homocysteine, vitamin A, vitamin D, vitamin E and metals, in the blood samples of participants selected using a case-control design were examined. Genomic DNA was extracted. FINDINGS TO DATE: The baseline population included 8045 preconception couples, 3054 single women and 15 615 early-pregnancy women. Data from 12 402 births were collected, and follow-up of the cohort for other outcomes is ongoing. Currently, 151 cases of CHD were identified after birth. The pilot analysis in a small subgroup showed that approximately 20.0% of preconception women and 44.9% of early-pregnancy women had red blood cell (RBC) folate levels that met the international recommendation for preventing neural tube defects. FUTURE PLANS: Once a sufficient number of CHD cases are achieved, we will investigate the quantitative association of preconception RBC folate levels with CHD using a nested case-control design. The SPCC will be followed up for 18 years to investigate extensive outcomes of growth, development, obesity, and common and rare diseases during childhood and adolescence according to our plan. Blood nutritional factors will be examined in participants selected for specific aims. The SPCC will also allow for prospective cohort studies on extensive research questions. TRIAL REGISTRATION NUMBER: NCT02737644.

Amido surface-functionalized magnetic molecularly imprinted polymers for the efficient extraction of Sibiskoside from Sibiraea angustata.

A general method for efficient and selective extraction of a target compound from complex natural products remains elusive, despite decades of research. By introducing a functional amido group on the surface of dispersity-enhanced magnetic nanoparticles, a nanoparticle receptor to selectively recognize Sibiskoside (a monoterpene) from the aerial portion of Sibiraea angustata by hydrogen bond interaction was synthesized. The superparamagnetic Fe3O4 nanoparticles were successively modified with tetraethyl orthosilicate (TEOS), amino and amido functional groups, and 4-vinylbenzoic acid (VBZA) was used as the functional monomer. A thin layer of poly (VBZA) imprinted with Sibiskoside was immobilized on the surface of magnetic carriers. Attributing to the amido group introduced into the magnetic particles, the template could attract and bind to the surface and promote the formation of a hydrogen bond system between the carrier, template molecules and functional monomer. High-density molecular recognition sites grew on the surface of magnetic substrate. The adsorption reached equilibrium at approximately 150 min, while fast adsorption occurred during the first 60 min. The maximum adsorption capacity has been found to be 13.75 mg g-1 according to calculation with the Langmuir isotherm. The selectivity coefficients of Molecular imprinting polypers (MIPs) for Sibiskoside with respect to Andrographolide, Loganin, Gastrodin, geraniol-1-O-[α-l-rhamnopyranosyl-(1 → 6)-1-ß-d-glucopyranoside] (GRG), Sibiscolacton and Sibiraic acid were 2.26, 1.43, 1.701.56, 1.05, 0.73 and, respectively. The results indicated that the MIPs possessed good specific adsorption capacity and selectivity toward Sibiskoside and had the potential to be a candidate for the separation and purification of monoterpenes from Sibiraea angustata, which is of great significance to obesity management.

Phosphorus removal from aqueous solution using a novel granular material developed from building waste.

In this study, a granular material (GM) developed from building waste was used for phosphate removal from phosphorus-containing wastewater. Batch experiments were executed to investigate the phosphate removal capacity of this material. The mechanism of removal proved to be a chemical precipitation process. The characteristics of the material and resulting precipitates, the kinetics of the precipitation and Ca2+ liberation processes, and the effects of dosage and pH were investigated. The phosphate precipitation and Ca2+ liberation processes were both well described by a pseudo-second-order kinetic model. A maximum precipitation capacity of 0.51 ± 0.06 mg g-1 and a liberation capacity of 6.79 ± 0.77 mg g-1 were measured under the experimental conditions. The processes reached equilibrium in 60 min. The initial solution pH strongly affected phosphate removal under extreme conditions (pH <4 and pH >10). The precipitates comprised hydroxyapatite and brushite. This novel GM can be considered a promising material for phosphate removal from wastewater.

Antibacterial and antifungal activities of various solvent extracts from the leaves and stem bark of Anisophyllea laurina R. Br ex Sabine used as traditional medicine in Guinea.

ETHNOPHARMACOLOGICAL RELEVANCE: Anisophyllea laurina R. Br ex Sabine is a plant that has been used in the folk medicine to treat malaria, dysentery, diabetes and toothache against bacterial infection. Through this study, most likely we are reporting for the first time, its effectiveness as an antibacterial and antifungal agent. AIM OF THE STUDY: To evaluate antibacterial and antifungal activities of various solvent extracts from the leaves and stem bark of A. laurina R. Br ex Sabine. MATERIALS AND METHODS: The various solvent extracts of leaves and stem bark were tested for antibacterial and antifungal activities against eight bacteria strains and three fungal strains using the well diffusion and micro-dilution methods. RESULTS: Antibacterial and antifungal activities of methanol and ethanol extracts were higher than the ethyl acetate and water extract. Leaves extracts had the highest potential activity against bacteria than stem bark extracts, but showed low antifungal activity compared to stem bark extracts. They also exhibited potent growth inhibitory activity against Pseudomonas aeruginosa, Salmonella typhimurium, Escherichia coli ATCC 8739 and Bacillus subtilis with MIC value of 125µg/mL. CONCLUSIONS: The current study confirmed that the leaves and stem bark extracts of A. laurina R. Br ex Sabine proved to be most effective as antibacterial and antifungal activities.

[Chemical constituents from the aerial part of Sibiraea angustata].

OBJECTIVE: To study the chemical constituents from the aerial part of Sibiraea angustata. METHODS: The constituents were isolated by various chromatographic techniques (HP-20 macroporous absorption resin, Sephadex LH-20 gel, RP-MPLC and PHPLC)and their structures were determined on the basis of physicochemical properties and their spectroscopic data,as well as literatures. RESULTS: Eleven compounds were separated and identified as p-methoxycinnamic acid(I), protocatechuic aldehyde(II), quercetin(III), isorhamnetin(IV), quercetin 3-O-beta-D-galactopyranoside (V),9-0-[beta-D-glucopyranoside]-3,4,5-trimethoxy cinnamyl alcohol(VI), syringaresinol-4'-O-beta-D-monoglucoside(VII), ntin(VIII), sibiraic acid(IX), sibiscolacton(X), methyl ferulic acid(XI). CONCLUSION: Compounds I-XIII are isolated from the genus of Sibiraea for the first time.

The R2R3-MYB, bHLH, WD40, and related transcription factors in flavonoid biosynthesis.

R2R3-MYB, bHLH, and WD40 proteins have been shown to control multiple enzymatic steps in the biosynthetic pathway responsible for the production of flavonoids, important secondary metabolites in Camellia sinensis. Few related transcription factor genes have been documented. The presence of R2R3-MYB, bHLH, and WD40 were statistically and bioinformatically analyzed on 127,094 C. sinensis transcriptome unigenes, resulting in identification of 73, 49, and 134 genes, respectively. C. sinensis phylogenetic trees were constructed for R2R3-MYB and bHLH proteins using previous Arabidopsis data and further divided into 27 subgroups (Sg) and 32 subfamilies. Motifs in some R2R3-MYB subgroups were redefined. Furthermore, Sg26 and Sg27 were expanded compared to Arabidopsis data, and bHLH proteins in C. sinensis were grouped into nine subfamilies. According to the functional annotation of Arabidopsis, flavonoid biosynthesis in C. sinensis was predicted to include R2R3-MYB genes in Sg4 (6), Sg5 (2), and Sg7 (1), as well as bHLH genes in subfamily 2 (2) and subfamily 24 (5). The wide evolutionary gap prevented phylogenetic analysis of WD40s; however, a single gene, CsWD40-1, was observed to share 80.4 % sequence homogeny with AtTTG1. Analysis of CsMYB4-1, CsMYB4-2, CsMYB4-3, CsMYB4-4, CsMYB5-1, and CsMYB5-2 revealed the interaction motif [DE]Lx2[RK]x3Lx6Lx3R, potentially contributing to the specificity of the bHLH partner in the stable MYB-bHLH complex. Full-length end-to-end polymerase chain reaction (PCR) and quantitative reverse transcriptase (qRT)-PCR were used to validate selected genes and generate relative expression ratio profiles in C. sinensis leaves by developmental stage and treatment conditions, including hormone and wound treatments. Potential target binding sites were predicted.

Differential gene expression in tea (Camellia sinensis L.) calli with different morphologies and catechin contents.

Tea (Camellia sinensis) is a commercially important crop that contains valuable secondary metabolites. To understand the molecular regulation of secondary metabolism in tea, we selected and analyzed two cell lines of tea callus (Yunjing63Y and Yunjing63X) that showed different morphological characteristics and catechin contents. Yunjing63Y callus was yellow and tight, while yunjing63X callus was white and loose. HPLC analyses showed that Yunjing63Y contained 3.71 times higher levels of catechins than Yunjing63X. Using cDNA amplified fragment-length polymorphism (cDNA-AFLP) we identified 68 genes that were differentially expressed between the two lines. Of the 68 differentially expressed ESTs, 40 showed higher expressions in Yunjing63Y and 28 showed higher expressions in Yunjing63X. BLASTX comparisons classified these ESTs into seven functional groups; phenylpropanoid metabolism (2.9%), UDPG-dependent glucosyl transferase (8.8%), transcription factors (11.8%), transporters (13.2%), signal transduction (19.1%), other metabolism (26.5%), and unknown (17.7%). We used qRT-PCR to validate the expression of genes and ESTs, and found that genes associated with flavan-3-ols biosynthesis and metabolism were expressed at higher levels in Yunjing63Y than in Yunjing63X. In addition, the expression of ESTs associated with flavonoid biosynthesis, regulation and transport were higher in Yunjing63Y than in Yunjing63X. The full-length cDNA of a EST coding for a putative MYB transcription factor was amplified using rapid amplification of cDNA ends (RACE). The resulting 1270 bp long cDNA, named CsMYB1, contained a 933-bp ORF encoding a 310-amino acid protein with a predicted molecular weight of 105.27 kDa and a predicted isoelectric point of 4.85 and showed highest homology to plant MYBs likely involved in stress signaling.

More tea for septic patients?--Green tea may reduce endotoxin-induced release of high mobility group box 1 and other pro-inflammatory cytokines.

Despite recent advances in antibiotic therapy and intensive care, sepsis remains widespread problems in critically ill patients. The high mortality of sepsis is in part mediated by bacterial endotoxin, which stimulates macrophages/monocytes to sequentially release early (e.g., TNF, IL-1, and IFN-gamma) and late (e.g., HMGB1) pro-inflammatory cytokines. In light of our recent discovery of HMGB1 as a late mediator of lethal systemic inflammation, and the observation that green tea (Camellia sinensis) dose-dependently attenuated bacterial endotoxin-induced HMGB1 release, we propose that regular tea intake might decrease the incidence of and mortality rates from lethal endotoxemia and sepsis.