Fibroblast growth factor 23 is associated with the development of gestational diabetes mellitus.

BACKGROUND: Besides its established impact on bone and mineral metabolism, it was suggested that fibroblast growth factor 23 (FGF23) might play an important role in the pathogenesis of type 2 diabetes. The impact of FGF23 on gestational diabetes mellitus (GDM), however, is not well understood. iFGF23 ELISAs measure the intact FGF23 molecule, whereas cFGF23 assays measure intact FGF23 as well as degradation products of FGF23. OBJECTIVES: The aim of this study is to compare the association of maternal and foetal cFGF23 and iFGF23 with GDM in a German birth cohort. METHODS: cFGF23 and iFGF23 were analysed in 826 random mother/child pairs from the Berlin Birth Cohort. RESULTS: Mothers who developed GDM had higher concentrations of iFGF-23 compared to mothers who did not suffer from GDM (19.73 vs. 13.23 pg/mL, p < 0.0001), but not higher concentrations of cFGF-23. Multivariant regression analyses showed that gestational diabetes is associated with iFGF23 independently of confounding factors such as age, BMI, ethnic background, family history of diabetes, smoking during pregnancy, and recurrent pregnancy loss. This, however, was only seen when using an iFGF23 ELISA measuring just the full length FGF23 and not in addition FGF23 fragments. No differences in both iFGF23 and cFGF23 concentrations between the GDM and non-GDM groups were detected in cord blood samples of the offspring. CONCLUSIONS: This study of a representative German birth cohort showed that maternal but not foetal iFGF23 is independently associated with GDM.

A Novel Punch Technique for Facial Pigmented Melanocytic Nevus.

The punch tool is a swift and practical instrument in the facial pigmented melanocytic nevus. However, few studies have evaluated the efficacy of the method for facial pigmented nevus. The aim of this study was to evaluate the practicability and effectiveness of removing facial pigmented nevus by punch biopsy technique. This was an observational study of patients with facial pigmented nevus in the Hospital of Plastic Surgery, Weifang Medical University. The ages of patients ranged from 15 to 36 years (average, 25 y). The outcome evaluations included Vancouver Scar Scale (VSS) score, esthetic appearance, and patient satisfaction. Following standard procedures, preoperative surgical excision was performed with safety margins. Anatomopathologic analysis of the surgical specimen was used as the gold standard to evaluate the accuracy of diagnosis by punch biopsy. From January 2019 to January 2020, this punch technique was carried out on 96 patients (151 pigmented nevus) with 35 melanocytic nevus on the forehead, 39 on the cheek, 21 on the eyelid, and 45 on the nose, whereas 11 were on nasolabial folds. The diameters of pigmented nevus are 0.5 to 10 mm on the face. All patients were evaluated at a follow-up visit ranging from 6 to 20 months (average, 11±1.5 mo) and healed with no complication. The histopathological examinations of the skin lesions showed benign outcomes. The mean Vancouver Scar Scale were 1.1±0.4. Ideal cosmetic and functional outcomes were achieved in 94 patients (97.9%). All patients achieved complete satisfaction except 2 patients with partial satisfaction. No recurrences and complications were recorded. This study demonstrated that the punch technique is an effective method to remove facial pigmented melanocytic nevus with acceptable functional and esthetic outcomes without relapse.

Inverse correlation of intact PTH, oxidized PTH as well as non-oxidized PTH with 25-hydroxyvitamin D3 in kidney transplant recipients.

Background: 25-hydroxyvitamin D (25(OH)D) and potentially also 1,25-dihydroxyvitamin D (1,25(OH)2D) inhibits the synthesis of parathyroid hormone (PTH) in the chief cells of the parathyroid gland. Clinical studies showing a negative correlation between (25(OH)D and PTH are in good agreement with these findings in basic science studies. However, PTH was measured in these studies with the currently clinically used 2nd or 3rd generation intact PTH (iPTH) assay systems. iPTH assays cannot distinguish between oxidized forms of PTH and non-oxidized PTH. Oxidized forms of PTH are the by far most abundant form of PTH in the circulation of patients with impaired kidney function. Oxidation of PTH causes a loss of function of PTH. Given that the clinical studies done so far were performed with an PTH assay systems that mainly detect oxidized forms of PTH, the real relationship between bioactive non-oxidized PTH and 25(OH)D as well as 1,25(OH)2D is still unknown. Methods: To address this topic, we compared for the first time the relationship between 25(OH)D as well as 1,25(OH)2D and iPTH, oxPTH as well as fully bioactive n-oxPTH in 531 stable kidney transplant recipients in the central clinical laboratories of the Charité. Samples were assessed either directly (iPTH) or after oxPTH (n-oxPTH) was removed using a column that used anti-human oxPTH monoclonal antibodies, a monoclonal rat/mouse parathyroid hormone antibody (MAB) was immobilized onto a column with 500 liters of plasma samples. Spearman correlation analysis and Multivariate linear regression were used to evaluate the correlations between the variables. Results: There was an inverse correlation between 25(OH)D and all forms of PTH, including oxPTH (iPTH: r=-0.197, p<0.0001; oxPTH: r=-0.203, p<0.0001; n-oxPTH: r=-0.146, p=0.001). No significant correlation was observed between 1,25(OH)2D and all forms of PTH. Multiple linear regression analysis considering age, PTH (iPTH, oxPTH and n-oxPTH), serum calcium, serum phosphor, serum creatinine, fibroblast growth factor 23 (FGF23), osteoprotegerin (OPG), albumin, and sclerostin as confounding factors confirmed these findings. Subgroup analysis showed that our results are not affected by sex and age. Conclusion: In our study, all forms of PTH are inversely correlated with 25-hydroxyvitamin D (25(OH)D). This finding would be in line with an inhibition of the synthesis of all forms of PTH (bioactive n-oxPTH and oxidized forms of PTH with minor or no bioactivity) in the chief cells of the parathyroid glad.

Multivesicular liposomes for the sustained release of thymopentin: stability, pharmacokinetics and pharmacodynamics.

The objective of the present study was to investigate the storage stability of thymopentin multivesicular liposomes (TP5-MVLs) prepared with different emulsifiers, and to study the pharmacokinetics and pharmacodynamics of the produced TP5-MVLs in vivo. The stability studies of TP5-MVLs indicated that MVLs particles prepared with mixed emulsifiers (Myrj52:solutolHS15 = 2:3) were stable at the storage temperature of 4 +/- 2 degrees C within 3 months. In addition, FITC-TP5-loaded MVLs was prepared for pharmacokinetic studies that after subcutaneous administration, the fluorescence signal lasted for about 5 days in plasma demonstrating that the rate of drug release from MVLs was very slow. The pharmacodynamic studies indicated that the therapeutic efficacy of TP5-MVLs after subcutaneous administration once every four days was the same as free TP5 solution after intravenous or subcutaneous administration once daily. In conclusion, MVLs, which possessed great storage stability, can be utilized to reduce the administration frequency of TP5, and therefore, served as a promising sustained release delivery system for polypeptide.

Head-to-Head Comparison of Oxidative Stress Biomarkers for All-Cause Mortality in Hemodialysis Patients.

Oxidative stress (OS) presents even in the early chronic kidney disease (CKD) stage and is exacerbated in patients with end-stage renal disease (ESRD) undergoing maintenance hemodialysis (MHD). There is still a debate over the association between oxidative stress and mortality. Our study aims to compare head-to-head the prognostic value of different oxidative markers for all-cause mortality in hemodialysis (HD) patients. We thus enrolled 347 patients on HD in this prospective study. Four OS biomarkers were measured (carbonyl proteins, myeloperoxidase (MPO), advanced oxidation protein products (AOPPs), and oxidized low-density lipoprotein (ox-LDL)). During the 60-month follow-up period, 9 patients have been lost to follow-up and 168 (48.4%) patients died. Concerning the oxidative stress (ox-stress) byproducts, carbonyl proteins were lower in survivors (105.40 ng/mL (IQR 81.30−147.85) versus 129.65 ng/mL (IQR 93.20−180.33); p < 0.001), with similar results for male patients (103.70 ng/mL (IQR 76.90−153.33) versus 134.55 ng/mL (IQR 93.95−178.68); p = 0.0014). However, there are no significant differences in MPO, AOPP, and ox-LDL between the two groups. Kaplan−Meier survival analysis indicated that patients in the higher carbonyl proteins concentration (>117.85 ng/mL group) had a significantly lower survival rate (log-rank test, p < 0.001). Univariate Cox regression analysis showed a positive correlation between carbonyl proteins and all-cause mortality in the higher and lower halves. Even after adjustment for conventional risk factors, it remained a statistically significant predictor of an increased risk of death in MHD. Univariate Cox regression analysis of MPO showed that continuous MPO and Log MPO were significantly associated with all-cause mortality, except for binary MPO (divided according to the median of MPO). Multivariate Cox analysis for MPO showed that the mortality prediction remains significant after adjusting for multiple factors. In conclusion, not all ox-stress biomarkers predict all-cause mortality in HD patients to a similar extent. In the present study, carbonyl proteins and MPO are independent predictors of all-cause mortality in HD patients, whereas AOPPs and oxLDL are clearly not associated with all-cause mortality in HD patients.

Empagliflozin reduces kidney fibrosis and improves kidney function by alternative macrophage activation in rats with 5/6-nephrectomy.

BACKGROUND: Sodium glucose cotransporter 2 (SGLT2) inhibitors originally developed for the treatment of type 2 diabetes are clinically very effective drugs halting chronic kidney disease progression. The underlying mechanisms are, however, not fully understood. METHODS: We generated single-cell transcriptomes of kidneys from rats with 5/6 nephrectomy before and after SGLT2 inhibitors treatment by single-cell RNA sequencing. FINDINGS: Empagliflozin treatment decreased BUN, creatinine and urinary albumin excretion compared to placebo by 39.8%, 34.1%, and 55%, respectively (p < 0.01 in all cases). Renal interstitial fibrosis and glomerulosclerosis was likewise decreased by 51% and 66.8%; respectively (p < 0.05 in all cases). 14 distinct kidney cell clusters could be identified by scRNA-seq. The polarization of M2 macrophages from state 1 (CD206-CD68- M2 macrophages) to state 5 (CD206+CD68+ M2 macrophages) was the main pro-fibrotic process, as CD206+CD68+ M2 macrophages highly expressed fibrosis-promoting genes and can convert into fibrocytes. Empagliflozin remarkably inhibited the expression of fibrosis-promoting (IFG1 and TREM2) and polarization-associated genes (GPNMB, LGALS3, PRDX5, and CTSB) in CD206+CD68+ M2 macrophages and attenuated inflammatory signals from CD8+ effector T cells. The inhibitory effect of empagliflozin on CD206+CD68+ M2 macrophages polarization was mainly achieved by affecting mitophagy and mTOR pathways. INTERPRETATION: We propose that the beneficial effects of empagliflozin on kidney function and morphology in 5/6 nephrectomyiced rats with established CKD are at least partially due to an inhibition of CD206+CD68+ M2 macrophage polarization by targeting mTOR and mitophagy pathways and attenuating inflammatory signals from CD8+ effector T cells. FUNDINGS: A full list of funding bodies that contributed to this study can be found in the Acknowledgements section.

[Knowledge on drinking water of adults in four cities of China].

OBJECTIVE: To understand the status on knowledge of drinking water among adults aged 18 - 60 yrs in Beijing, Shanghai, Chengdu and Guangzhou of China. METHODS: A total of 1483 adults aged 18 - 60 yrs from the four cities (Beijing, Shanghai, Chengdu and Guangzhou) were selected using multiple-stage random sampling method. The information was collected by face-to-face interview. Through face to face interview, investigators surveyed the daily water intake, the harm of dehydration, time of drinking water and healthy drinks knowledge using a questionnaire. Knowledge awareness was compared through χ(2) test for different cities, urban and rural, genders. RESULTS: The unawareness rate of minimum drinking water volume (1200 ml/d) was 28.4% (421/1483). The rate in Chengdu was the highest (41.8%, 156/373). The rate in women (32.4%, 241/745) was significantly higher than that in men (24.4%, 180/738) (χ(2) = 11.55, P < 0.01). The unawareness rate of the harm of dehydration was 14.4% (213/1483). The rate in Chengdu (18.5%, 69/373) and Guangzhou (18.3%, 68/371) was higher than that in the other cities. The rate in rural (18.6%, 137/736) was significantly higher than that in urban (10.2%, 76/747) (χ(2) = 21.47, P < 0.01). The people who considered drinking water when they felt thirsty as healthy behavior accounted for 48.3% (716/1483). The rate in Chengdu was the highest (59.0%, 220/373). The unawareness rate of plain water being the best accounted for 18.9% (280/1483), while 17.9% (50/280) of them considered beverages as the best. CONCLUSIONS: The knowledge of drinking water among 18 - 60 yrs residents in the four cities is not comprehensive. We should enhance propaganda of healthy drinking water.

[Water intake of adults in four cities of China in summer].

OBJECTIVES: To investigate total drinking water intake of adults in the four cities of China in summer. METHODS: A total of 1483 adults aged 18 - 60 yrs from Beijing, Shanghai, Chengdu and Guangzhou were selected using multiple-stage random sampling method. The information of amounts and types of daily drinking water was recorded by subjects for seven consecutive days using a quantitative measurement. The amounts and types of daily drinking water among different cities and between men and women or urban and rural was analyzed. RESULTS: The median of daily total drinking water of subjects was 1488 ml, with significant difference among the four cities (1579, 1793, 1150, 1467 ml in Beijing, Shanghai, Chengdu and Guangzhou city, respectively, χ(2) = 154.31, P = 0.000). The median of daily drinking water was significantly higher in men (1679 ml) than women (1370 ml) (Z = 8.34, P = 0.000), but no significant difference was found between urban (1514 ml) and rural (1466 ml) daily drinking water median (Z = -0.81, P = 0.420). The median of daily consumption of plain water, tea and beverages were 786, 109, 186 ml, respectively. Among four cities, the highest consumption of plain water was in subjects of Guangzhou (917 ml), while the highest tea consumption in Shanghai (257 ml) and the highest beverages consumption in Shanghai (323 ml) and Beijing (264 ml). Consumption of tea in men (229 ml) was significantly higher than that in women (57 ml) (Z = 7.52, P = 0.000). Subjects in urban (693 ml) had lower consumption of plain water than those in rural (914 ml). The proportion was 32.4% (480/1483) for subjects with water drinking less than 1200 ml per day. CONCLUSION: The daily consumption of total drinking water, including plain water, tea and beverages is different in adults among different cities and is different in gender and regions. It is nearly 1/3 of subjects with daily total drinking water less than current Chinese recommended water intake (1200 ml).

[Food water sources analysis of adults in four cities of China in summer].

OBJECTIVE: To investigate the food water sources of adults in the four cities of China in summer. METHODS: A total of 64 adults aged 18 - 60 yrs from four cities (Beijing, Shanghai, Chengdu and Guangzhou)were selected using convenient sampling method. The food samples were collected by using duplicate portion method and the water content of food samples were determined by using the national standard. The information on amounts and types of daily drinking soup was recorded by subjects using a quantitative measurement. RESULTS: A total of 63 subjects had completed the investigation. The median of daily water from food of subjects was 1157 ml, while water from staple food was 480 ml, the median ratio of which accounting for 41.8% of water from food. Water from dishes was 427 ml, the median ratio of which accounting for 37.9% of water from food. Water from soups was 133 ml, the median ratio of which accounting for 13.8% of water from food. Water from snacks was 0 ml, the median ratio of which accounting for 6.4% of water from food. The medians of water from staple food (384, 388 and 435 ml, respectively) in Beijing, Shanghai and Guangzhou was significant lower than that of Chengdu subjects' (900 ml, χ(2) = 21.27, P = 0.000). But the median proportion of water from staple food of subjects in Beijing (47.7%) was significantly higher than that in Chengdu (43.2%), Shanghai (42.9%) and Guangzhou (33.9%) (χ(2) = 8.69, P = 0.034). The median of amount and proportion of water from soups of subjects in Guangzhou (267 ml, 24.4%) and Chengdu(278 ml, 15.7%) was significantly higher than that in Shanghai (133 ml, 9.0%) and Beijing (100 ml, 5.8%) (amount: χ(2) = 22.52, P = 0.000;proportion: χ(2) = 16.27, P = 0.001). CONCLUSION: The staple food and dishes are the main sources of daily food water.

[Water intake sources analysis of adults in four cities of China in summer].

OBJECTIVE: To investigate the water intake sources of adults in the four cities of China in summer. METHODS: A total of 64 adults aged 18 - 60 yrs from the four cities (Beijing, Shanghai, Chengdu and Guangzhou) were selected using convenient sampling method. The information on amounts and types of daily drinking water was recorded by subjects for three consecutive days using a quantitative measurement. The food samples were collected by using duplicate portion method and the water content of food samples were determined. RESULTS: The median of daily total water intake of 63 subjects was 3045 ml, while daily total drinking water was 1600 ml, the median ratio of which accounting for 55.8% of total water intake. Water from food was 1157 ml, the median ratio of which accounting for 41.4% of total water intake. The median of daily total water intake (3566 ml) and the ratio of water from food (55.3%) of subjects in Chengdu was significantly higher than subjects in Guangzhou (2929 ml, 45.4%), Shanghai (2748 ml, 33.1%) and Beijing (2743 ml, 31.3%) (daily total water intake: χ² = 10.42, P = 0.015;the proportion of water: χ² = 28.48, P < 0.01). Daily total water intake in men (3302 ml) was significantly higher than that in women (2900 ml) (Z = 2.35, P = 0.019). CONCLUSION: Daily total water intake of subjects in summer was more than the current Chinese recommended total water intake (2200 ml), with the main part of drinking water and water from food.

Potential gene flow of two herbicide-tolerant transgenes from oilseed rape to wild B. juncea var. gracilis.

Four successive reciprocal backcrosses between F(1) (obtained from wild Brassica juncea as maternal plants and transgenic glyphosate- or glufosinate-tolerant oilseed rape, B. napus, as paternal plants) or subsequent herbicide-tolerant backcross progenies and wild B. juncea were achieved by hand pollination to assess potential transgene flow. The third and forth reciprocal backcrosses produced a number of seeds per silique similar to that of self-pollinated wild B. juncea, except in plants with glufosinate-tolerant backcross progeny used as maternal plants and wild B. juncea as paternal plants, which produced fewer seeds per silique than did self-pollinated wild B. juncea. Germination percentages of reciprocal backcross progenies were high and equivalent to those of wild B. juncea. The herbicide-tolerant first reciprocal backcross progenies produced fewer siliques per plant than did wild B. juncea, but the herbicide-tolerant second or third reciprocal backcross progenies did not differ from the wild B. juncea in siliques per plant. The herbicide-tolerant second and third reciprocal backcross progenies produced an amount of seeds per silique similar to that of wild B. juncea except for with the glufosinate-tolerant first and second backcross progeny used as maternal plants and wild B. juncea as paternal plants. In the presence of herbicide selection pressure, inheritance of the glyphosate-tolerant transgene was stable across the second and third backcross generation, whereas the glufosinate-tolerant transgene was maintained, despite a lack of stabilized introgression. The occurrence of fertile, transgenic weed-like plants after only three crosses (F(1), first backcross, second backcross) suggests a potential rapid spread of transgenes from oilseed rape into its wild relative wild B. juncea. Transgene flow from glyphosate-tolerant oilseed rape might be easier than that from glufosinate-tolerant oilseed rape to wild B. juncea. The original insertion site of the transgene could affect introgression.

Mechanisms of phospholipid complex loaded nanoparticles enhancing the oral bioavailability.

The purpose of the present study was to study the mechanisms of salvianolic acid B phospholipid complex loaded nanoparticles (SalB-PLC-NPs) enhancing the oral bioavailability of SalB by in situ perfusion model in rats and to evaluate the potential of phospholipid complex loaded nanoparticles (PLC-NPs) serving as an efficient oral delivery system to enhance the bioavailability of highly water-soluble drugs. SalB-PLC-NPs, prepared by a solvent evaporation method, exhibited a spherical shape with a mean particle size and a zeta potential of 112.2 nm and -44.2 mV, respectively. The drug entrapment efficiency and drug loading were 86.19% and 3.21%, respectively. The lyophilized SalB-PLC-NPs, prepared with 10% maltose as the cryoprotectant, presented sustained release profiles in artificial gastric juice (0.1 M HCl with pH 1.2) and intestinal juice (PBS with pH 6.8 and 7.4). The absorption mechanisms were studied using a modified in situ perfusion method in rats, which showed the segment dependent absorption characteristics of SalB, SalB-PLC as well as SalB-PLC-NPs. The greatest absorption was obtained when SalB-PLC-NPs were perfused in colon. The possibility of intestinal lymphatic transport of SalB-PLC-NPs was investigated using mesenteric lymph vessel cannulation. Microscope (fluorescence and natural light) observation of lymph indicated that nanoparticles underwent intestinal lymphatic transport. In conclusion, the enhanced oral bioavailability of SalB was contributed to both the PLC and NPs. Importantly, our studies indicate that PLC-NPs may be a promising delivery system to enhance the oral bioavailability of highly water-soluble drugs.

Inhalable microparticles as carriers for pulmonary delivery of thymopentin-loaded solid lipid nanoparticles.

PURPOSE: Microparticles containing solid lipid nanoparticles (SLNs) are receiving increased attention as carriers for the lung delivery of the SLNs. Thus, we aim to prepare the hybrid microparticles and thoroughly evaluate their feasibility for the pulmonary drug delivery. METHODS: The microparticles were prepared by co-spray-drying the thymopentin (TP5)-loaded SLNs with bulking agents. Thereafter, we systematically estimated the potential of the microparticles as the carriers for the pulmonary delivery of the SLNs, including the investigations of their characteristics, aerodynamic properties, pharmacokinetics and pharmacodynamics. RESULTS: The spherical and hollow microparticles presented a size of 4.1 +/- 0.1 microm and a low tap density of 0.175 +/- 0.02 g/cm(3). In addition, the microparticles showed a high aerosolization efficiency (emitted dose of 98.0% +/- 1.23% and respirable fraction of 51.07% +/- 1.21%). Furthermore, the SLNs could be easily recovered from the microparticles without essential changes on their characteristics and the drug release behavior. The pharmacokinetic and pharmacodynamic studies suggested that, compared to i.v. TP5 solution, the bioavailability and therapeutic efficacy of TP5 were remarkably strengthened after the pulmonary administration of the microparticles. CONCLUSIONS: Taken together, we believe the microparticles were suitable for inhalation and possessed an ample potential for the pulmonary delivery of the SLNs.

Development of rat urinary HPLC-UV profiling for metabonomic study on Liuwei Dihuang Pills.

Since urine contains thousands of metabolites, there are no universal analytical techniques to analyze these compounds simultaneously. Therefore, a readily obtained and simple sample pretreatment high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection urinary profiling with good reproducibility was developed for metabonomic study in this paper. The results of current study clearly showed that there were different phenotypes of metabolites based on HPLC-UV urinary profiling after administration of Liuwei Dihuang Pills (LWPs) or Carrageenan-stimulated inflammation model, and those could be conveniently discriminated by principal component analysis (PCA). In addition, the results also showed that LWPs could restore the metabolite network that disturbed by inflammation, which would be a proof of therapeutic efficacy of LWPs to inflammation by metabonomics study. Our study indicated that HPLC-UV method could be used as a potentially powerful tool for urinary profiling on metabonomic studies.

Enhanced oral bioavailability of salvianolic acid B by phospholipid complex loaded nanoparticles.

With a simple and feasible method, a salvianolic acid B-phospholipid complex was prepared to increase the absorption of salvianolic acid B in the gastrointestinal (GI) tract. Because the solubility of the complex in water was very poor, the complex was encapsulated into nanoparticles to facilitate its administration. The physicochemical properties of the complex were investigated by differential scanning calorimetry (DSC), infrared scanning, ultraviolet scanning and X-ray diffraction (XRD), and the solubility of salvianolic acid B and the complex in water or n-octanol was measured. The pharmacokinetic characteristics and bioavailability were compared after oral administration of salvianolic acid B (500 mg/kg) and the complex nanoparticles (450 mg/kg equivalent to salvianolic acid B). The results proved the drastic decrease in the solubility of salvianolic acid B in water after successful formation of the drug-phospholipid complex. After oral administration the peak plasma concentration (Cmax) of salvianolic acid B given by the complex nanoparticles was 3.4 microg/ml which was much higher than that of salvianolic acid B sample (Cmax = 0.9 microg/ml), with Tmax of 75 min, 45 min and AUC of 664, 257 microg/ml min, respectively. The relative bioavailability (F) reached 286%.

Use of RNAi technology to develop a PRSV-resistant transgenic papaya.

Papaya ringspot virus (PRSV) seriously limits papaya (Carica papaya L.) production in tropical and subtropical areas throughout the world. Coat protein (CP)- transgenic papaya lines resistant to PRSV isolates in the sequence-homology-dependent manner have been developed in the U.S.A. and Taiwan. A previous investigation revealed that genetic divergence among Hainan isolates of PRSV has allowed the virus to overcome the CP-mediated transgenic resistance. In this study, we designed a comprehensive RNAi strategy targeting the conserved domain of the PRSV CP gene to develop a broader-spectrum transgenic resistance to the Hainan PRSV isolates. We used an optimized particle-bombardment transformation system to produce RNAi-CP-transgenic papaya lines. Southern blot analysis and Droplet Digital PCR revealed that line 474 contained a single transgene insert. Challenging this line with different viruses (PRSV I, II and III subgroup) under greenhouse conditions validated the transgenic resistance of line 474 to the Hainan isolates. Northern blot analysis detected the siRNAs products in virus-free transgenic papaya tissue culture seedlings. The siRNAs also accumulated in PRSV infected transgenic papaya lines. Our results indicated that this transgenic papaya line has a useful application against PRSV in the major growing area of Hainan, China.

Publisher Correction: Use of RNAi technology to develop a PRSV-resistant transgenic papaya.

A correction to this article has been published and is linked from the HTML version of this paper. The error has been fixed in the paper.

A Comprehensive Assessment of the Effects of Transgenic Cry1Ac/Cry1Ab Rice Huahui 1 on Adult Micraspis discolor (Fabricius) (Coleoptera: Coccinellidae).

Micraspis discolor (Fabricius) (Coleoptera: Coccinellidae) is a widely distributed coleoptera predator in southern Asia in rice ecosystem, and adult M. discolor feed on both rice pollen and soft-bodied arthropods. Bitrophic bioassay and tritrophic bioassay were conducted to evaluate the potential impact of Cry1Ac/Cry1Ab-expressing rice Huahui 1 and its non-transgenic counterpart Minghui 63 on fitness parameters of adult M. discolor. The results showed that the survival, and fecundity of this beetle' adults were not different when they fed on Bt rice or non-Bt rice pollen or Nilaparvata lugens (Stål) reared on Bt rice or non-Bt rice. Toxicity assessment to ensure M. discolor adults were not sensitive to Cry1Ab or Cry1Ac protein independent from the pollen background, M. discolor adults were fed with an artificial diet containing Cry1Ac, Cry1Ab or both protein approximately 10 times higher concentration than in Huahui 1 rice pollen. No difference was detected for any of the life-table parameters tested between Cry protein-containing and pure diet. Artificial diet containing E-64 (N-(trans-Epoxysuccinyl)-L-leucine 4-guanidinobutylamide) was included as a positive control. In contrast, the pre-oviposition and fecundity of M. discolor were significantly adversely affected by feeding on E-64-containing diet. In both bioassays, the uptakes of Cry protein by adult M. discolor were tested by ELISA measurements. These results indicated that adults of M. discolor are not affected by Cry1Ab- or Cry1Ac-expressing rice pollen and are not sensitive to Cry protein at concentrations exceeding the levels in rice pollen in Huahui1. This suggests that M. discolor adults would not be harmed by Cry1Ac/Cry1Ab rice if Bt rice Huahui 1 were commercialized.

Genome-wide characterization and analysis of bZIP transcription factor gene family related to abiotic stress in cassava.

The basic leucine zipper (bZIP) transcription factor family plays crucial roles in various aspects of biological processes. Currently, no information is available regarding the bZIP family in the important tropical crop cassava. Herein, 77 bZIP genes were identified from cassava. Evolutionary analysis indicated that MebZIPs could be divided into 10 subfamilies, which was further supported by conserved motif and gene structure analyses. Global expression analysis suggested that MebZIPs showed similar or distinct expression patterns in different tissues between cultivated variety and wild subspecies. Transcriptome analysis of three cassava genotypes revealed that many MebZIP genes were activated by drought in the root of W14 subspecies, indicating the involvement of these genes in the strong resistance of cassava to drought. Expression analysis of selected MebZIP genes in response to osmotic, salt, cold, ABA, and H2O2 suggested that they might participate in distinct signaling pathways. Our systematic analysis of MebZIPs reveals constitutive, tissue-specific and abiotic stress-responsive candidate MebZIP genes for further functional characterization in planta, yields new insights into transcriptional regulation of MebZIP genes, and lays a foundation for understanding of bZIP-mediated abiotic stress response.