[Evaluation of Heavy Metal Distribution Characteristics and Ecological Risk of Soil of Vegetable Land for Hong Kong in Ningxia].

Heavy metal pollution in farmland soil can affect the growth, development, and yield of vegetable crops, as well as the quality and taste of vegetables, and can be continuously transmitted and enriched through the food chain, which ultimately poses a certain hazard to human health in the long term. Therefore, in order to investigate the distribution characteristics of soil heavy metals after years of multi-crop planting of vegetables supplied to Hong Kong, predict their ecological risks, and analyze the causes of pollution formation, 477 surface soil samples of vegetable fields supplied to Hong Kong in Ningxia were collected for three consecutive years from 2019 to 2021, and the contents and distribution characteristics of eight heavy metals, namely, As, Cd, Cr, Hg, Pb, Cu, Zn, and Ni were analyzed. The soil heavy metal pollution status of vegetable fields supplied to Hong Kong in Ningxia was evaluated using the single-factor pollution index method, Nemero's comprehensive pollution index method, land accumulation index method, and potential ecological risk index method, and the sources of heavy metals in vegetable fields supplied to Hong Kong in Ningxia were analyzed using the Pearson's correlation analysis and the principal component analysis method. The results showed that the mean values of As, Cd, Cr, Hg, Cu, and Zn in the soils of Ningxia's vegetable fields were higher than the background values of Ningxia soils, but the contents of all eight heavy metals were lower than the risk screening values of domestic agricultural soils; in terms of spatial distribution, As, Cr, and Ni showed contiguous high values in the northwestern, central, and southern parts of the study area, whereas Pb, Zn, Cd, Hg, and Cu showed high values in the northwestern and southern parts of the study area. The single-factor index method and the Nemero's comprehensive pollution index method showed that the soil of Ningxia's vegetable farmland for Hong Kong was at the clean level as a whole. The results of the ground accumulation index method showed that the pollution in the study area was mainly Hg and Cd pollution, and the pollution areas were mainly concentrated in the northwest and south of the study area. The potential ecological risk index showed that Hg and Cd were the main risk elements, among which Hg was dominated by moderate, strong, and very strong ecological risks, accounting for 44.65 %, 44.65 %, and 1.26 %, respectively, and Cd was dominated by moderate and strong risks, accounting for 65.83 % and 3.56 %. The comprehensive Pearson correlation analysis and principal component analysis showed that the pollution sources of eight heavy metals could be divided into three categories, namely, natural sources:Cu, Zn, Pb, As, Ni, and Cr; agricultural sources:Cd; and industrial and agricultural sources:Hg. From a comprehensive point of view, the heavy metals of the soil in the fields of vegetables supplied to Hong Kong had not exceeded the standard, and the environmental conditions of the soil were good, such that the production of vegetables supplied to Hong Kong by Ningxia was at a safe level overall. The results of the study can provide a theoretical basis for the safe utilization of soil in vegetable fields and the green production of vegetables supplied to Hong Kong in Ningxia, which were aimed to provide help for the safe production of vegetable fields supplied to Hong Kong, the rational application of fertilizers, agronomic planning, and the adjustment of planting structure.

A machine learning approach for predicting radiation-induced hypothyroidism in patients with nasopharyngeal carcinoma undergoing tomotherapy.

The purpose of this study was to establish an integrated predictive model that combines clinical features, DVH, radiomics, and dosiomics features to predict RIHT in patients receiving tomotherapy for nasopharyngeal carcinoma. Data from 219 patients with nasopharyngeal carcinoma were randomly divided into a training cohort (n = 175) and a test cohort (n = 44) in an 8:2 ratio. RIHT is defined as serum thyroid-stimulating hormone (TSH) greater than 5.6 µU/mL, with or without a decrease in free thyroxine (FT4). Clinical features, 27 DVH features, 107 radiomics features and 107 dosiomics features were extracted for each case and included in the model construction. The least absolute shrinkage and selection operator (LASSO) regression method was used to select the most relevant features. The eXtreme Gradient Boosting (XGBoost) was then employed to train separate models using the selected features from clinical, DVH, radiomics and dosiomics data. Finally, a combined model incorporating all features was developed. The models were evaluated using receiver operating characteristic (ROC) curves and decision curve analysis. In the test cohort, the area under the receiver operating characteristic curve (AUC) for the clinical, DVH, radiomics, dosiomics and combined models were 0.798 (95% confidence interval [CI], 0.656-0.941), 0.673 (0.512-0.834), 0.714 (0.555-0.873), 0.698 (0.530-0.848) and 0.842 (0.724-0.960), respectively. The combined model exhibited higher AUC values compared to other models. The decision curve analysis demonstrated that the combined model had superior clinical utility within the threshold probability range of 1% to 79% when compared to the other models. This study has successfully developed a predictive model that combines multiple features. The performance of the combined model is superior to that of single-feature models, allowing for early prediction of RIHT in patients with nasopharyngeal carcinoma after tomotherapy.

[Risk Assessment and Sources of Heavy Metals in Farmland Soils of Yellow River Irrigation Area of Ningxia].

The farmland environment is directly related to the quality and safety of agricultural products. In order to understand the characteristics and main influencing factors of heavy metals in farmland soil in the Yellow River irrigation area of Ningxia, sampling and monitoring were conducted for five consecutive years from 2017 to 2021, and the distribution characteristics and correlation of heavy metals were analyzed. The pollution status and potential ecological risks of heavy metals were evaluated, and the main sources of heavy metals in farmland were analyzed. The results showed that the average values of Pb, As, Zn, Ni, Cu, Hg, Cr, and Cd in the soil of the Ningxia Yellow River irrigation area were 19.74, 11.67, 66.88, 29.09, 22.55, 0.03, 62.27, and 0.19 mg·kg-1, respectively, which were enriched to some extent compared with the background values of the soil environment in Ningxia. Among them, Hg and Cd had middle- and high-grade ecological risk points; however, none of them exceeded the control value of agricultural land soil pollution risk, and all sampling sites had no high-risk or extremely high-risk levels. The results of source analysis based on positive matrix factorization (PMF) and correlation analysis showed that there were five main sources of heavy metals in farmland soil in the study area: natural sources, mixed sources of industrial and mining activities and the production and life of residents, transportation sources, agricultural production activities sources, and industrial sources, with contribution rates of 26.54%, 25.59%, 22.52%, 15.63%, and 9.72%, respectively. On the whole, the heavy metals in farmland soil in the Ningxia Yellow River irrigation area did not exceed the standard, and there was no high-level ecological risk. The production environment of the farmland soil was good, but the contribution rate of human activities to soil heavy metals was large.

Facile Synthesis of Uniform Mesoporous Nb2O5 Micro-Flowers for Enhancing Photodegradation of Methyl Orange.

The removal of organic pollutants using green environmental photocatalytic degradation techniques urgently need high-performance catalysts. In this work, a facile one-step hydrothermal technique has been successfully applied to synthesize a Nb2O5 photocatalyst with uniform micro-flower structure for the degradation of methyl orange (MO) under UV irradiation. These nanocatalysts are characterized by transmission and scanning electron microscopies (TEM and SEM), X-ray diffraction (XRD), Brunauer-Emmett-Teller (BET) method, and UV-Vis diffuse reflectance spectroscopy (DRS). It is found that the prepared Nb2O5 micro-flowers presents a good crystal phases and consist of 3D hierarchical nanosheets with 400-500 nm in diameter. The surface area is as large as 48.6 m2 g-1. Importantly, the Nb2O5 micro-flowers exhibit superior catalytic activity up to 99.9% for the photodegradation of MO within 20 mins, which is about 60-fold and 4-fold larger than that of without catalysts (W/O) and commercial TiO2 (P25) sample, respectively. This excellent performance may be attributed to 3D porous structure with abundant catalytic active sites.

Study on adsorption of ammonia nitrogen by iron-loaded activated carbon from low temperature wastewater.

In order to improve the adsorption efficiency of ammonia nitrogen in low temperature wastewater, the modified activated carbon (Fe-AC) was prepared by impregnation-calcination modification of Fe(NO3)3. The characterization results indicated that the total pore volume, specific surface area and the point of zero charge of activated carbon increased after modification. A better adsorption effect was achieved under neutral condition than under alkaline or acidic condition. The effect of Ca2+ on competitive adsorption of NH4+ was greater than that of Na+ when both cations were present. Pseudo-first-order kinetic model was confirmed to be consistent with Fe-AC adsorption kinetic data, and Langmuir model was consistent with adsorption isotherm data. The adsorption thermodynamics demonstrated that the ammonia nitrogen adsorption process by Fe-AC was spontaneous and low-temperature was helpful to improve the adsorption capacity. The mechanism of adsorption of ammonia nitrogen by Fe-AC was the comprehensive effect of physical adsorption and chemical adsorption, which was the essential reason for improving the adsorption efficiency of ammonia nitrogen by Fe-AC at a low temperature. This research offered a new way for the modification of activated carbon and a new method for the removal of ammonia nitrogen at a low temperature.

[Development of critical nitrogen dilution curve based on leaf dry matter for maize under drip irrigation]. / 基于叶片干物质的滴灌玉米临界氮稀释曲线构建.

To provide a theoretical reference to nitrogen fertilizer management for maize and eco-enviromental protection in the irrigated area of Ningxia, we established the critical nitrogen concentration (Nc) dilution curve of maize under drip-irrigated condition. The nitrogen nutrition index (NNI) was deduced from the established Nc curve model, to diagnose feasibility of nitrogen nutrition. Six levels of nitrogen application rates including 0, 90, 180, 270, 360 and 450 kg·hm-2 were conducted in field with maize variety 'Tianci 19'. During two-year field study, the Nc dilution curve and NNI model of maize were established based on leaf dry matter (LDM). The results showed that: 1) There was a negative power function relationship between LDM and Nc, which presented as two parts, if LDM<1.15 t·hm-2, Nc=3.2%, while if LDM≥1.15 t·hm-2, Nc=3.29LDM-0.29. 2) The root mean square error (RMSE) and standardized root mean square error (n-RMSE) of the model were 0.203 and 8.0%, respectively, with fine stability among years. 3) The NNI ranged from 0.47 to 1.44 with different nitrogen application rates. In addition, NNI was positively correlated with yield and negatively correlated with agronomic utilization efficiency of nitrogen fertilizer at different growth stages of maize. Therefore, NNI could be used to explain maize yield changes with and without nitrogen limit under drip-irrigated system. In conclusion, the critical nitrogen dilution curve established from leaf dry matter could be applied to accurately estimate nitrogen nutrition status throughout the whole growing stages from the jointing stage to silking stage of maize under drip irrigated system.

[Correlation between fractional exhaled nitric oxide and airway reversibility in children with IgE-mediated asthma].

OBJECTIVE: To study the correlation between fractional exhaled nitric oxide (FeNO) and airway reversibility in children with IgE-mediated asthma. METHODS: A total of 86 children, aged 6-14 years, who were initially diagnosed with acute attack of asthma from September 2016 to August 2018 were enrolled as subjects. According to the results of serum specific IgE, they were divided into IgE mediated group with 61 children and non-IgE mediated group with 25 children. According to the results of allergen detection, the IgE mediated group was further divided into four groups with one, two, three, and four or more positive allergens. FeNO and the parameters of pulmonary ventilation function before and after dilation test were measured. Pearson correlation analysis was used to evaluate the correlation of FeNO with each parameter of pulmonary function. RESULTS: The IgE mediated group had significantly higher FeNO than the non-IgE mediated group (P<0.05). FeNO increased with the increase in the number of positive serum specific allergens (P<0.05). In the IgE mediated group, FeNO level was positively correlated with the change in forced expiratory volume in the first second (FEV1) and the improvement in percentage of predicted FEV1 after medication in bronchial dilation test (r=0.655 and 0.473 respectively, P<0.05). The FeNO level was not correlated with FEV1, percentage of predicted FEV1, peak expiratory flow (PEF), change in PEF after medication, percentage of predicted PEF (PEF%pred), and improvement in PEF%pred after medication (P>0.05). In the non-IgE mediated group, FeNO level was not correlated with the above indicators (P>0.05). CONCLUSIONS: FeNO level is associated with the degree of allergies. For children with IgE-mediated asthma, FeNO is positively correlated with airway reversibility, which has a certain value in the diagnosis of asthma, disease evaluation, and understanding of airway reversibility. For children with non-IgE-mediated asthma, FeNO cannot be used to evaluate airway reversibility. These two types of asthma should be treated differently.

Correlation between fractional exhaled nitric oxide and airway reversibility in children with IgE-mediated asthma / 中国当代儿科杂志

OBJECTIVE@#To study the correlation between fractional exhaled nitric oxide (FeNO) and airway reversibility in children with IgE-mediated asthma.@*METHODS@#A total of 86 children, aged 6-14 years, who were initially diagnosed with acute attack of asthma from September 2016 to August 2018 were enrolled as subjects. According to the results of serum specific IgE, they were divided into IgE mediated group with 61 children and non-IgE mediated group with 25 children. According to the results of allergen detection, the IgE mediated group was further divided into four groups with one, two, three, and four or more positive allergens. FeNO and the parameters of pulmonary ventilation function before and after dilation test were measured. Pearson correlation analysis was used to evaluate the correlation of FeNO with each parameter of pulmonary function.@*RESULTS@#The IgE mediated group had significantly higher FeNO than the non-IgE mediated group (P0.05). In the non-IgE mediated group, FeNO level was not correlated with the above indicators (P>0.05).@*CONCLUSIONS@#FeNO level is associated with the degree of allergies. For children with IgE-mediated asthma, FeNO is positively correlated with airway reversibility, which has a certain value in the diagnosis of asthma, disease evaluation, and understanding of airway reversibility. For children with non-IgE-mediated asthma, FeNO cannot be used to evaluate airway reversibility. These two types of asthma should be treated differently.

MicroRNA-221 sponge therapy attenuates neointimal hyperplasia and improves blood flows in vein grafts.

BACKGROUND: Vein graft failure due to neointimal hyperplasia remains an important and unresolved problem of cardiovascular surgery. MicroRNA-221 (miR-221) has been shown to play a major role in regulating vascular smooth muscle cell (VSMC) proliferation and phenotype transformation. Thus, the purpose of this study is to determine whether adenovirus mediated miR-221 sponge gene therapy could inhibit vein graft neointimal hyperplasia. METHODS: Adenovirus encoding miR-221 sponge (Ad-miR-221-SP) was used to inhibit VSMC proliferation in vitro and neointimal formation in vivo. Expression of miRNA-221 was evaluated in cultured VSMC and in rat vein graft models following transduction with Ad-miR-221-SP, Ad-Control-SP (without miR-221 antisense binding sites), or Ad-GFP (control). To accelerate the transfer of miR-221 sponge gene to the vein grafts, 20% poloxamer F-127 gel was used to extend virus contact time and 0.25% trypsin to increase virus penetration. RESULTS: miR-221 sponges can significantly decrease the expression of miR-221 and proliferation in cultured VSMC. Cellular proliferation rates were significantly reduced in miR-221 sponge treated grafts as compared with controls at 6 weeks after bypass surgery (19.8% versus 43.6%, P=0.0028). miR-221 sponge gene transfer reduced the neointimal area (210.75 ± 24.13 versus 67.01 ± 12.02, P<0.0001), neointimal thickness (171.86 ± 27.87 versus 64.13 ± 16.23, P<0.0001) and neointima/media ratio (0.74 ± 0.21 versus 1.95 ± 0.25, P<0.0001) in vein grafts versus controls. miR-21 sponge treatment was also improved hemodynamics in vein grafts. We have further identified that p27 (Kip1) is a potential target gene of miR-221 in vein grafts. CONCLUSION: miR-221 sponge therapy can significantly reduce miR-221 activity and inhibit neointimal hyperplasia in vein grafts. Locally adventitial delivery of adenoviruses mediated miRNA sponges may be promising gene therapies to prevent vein graft failure.

[Dynamics simulation on plant growth, N accumulation and utilization of processing tomato at different N fertilization rates].

Three field experiments were conducted to simulate the dynamics of aboveground biomass, N accumulation and utilization of drip-irrigated processing tomatoes at different N fertilization rates (0, 75, 150, 300, 450, or 600 kg x hm(-2)). The results showed that Logistic models best described the changes in aboveground biomass, N accumulation, and utilization of accumulated N efficiency with the physiological development time (PDT). Rapid accumulation of N began about 4-6 d (PDT) earlier than the rapid accumulation of aboveground biomass. The momentary utilization rate of N (NMUR) increased after emergence, reached a single peak, and then decreased. The N accumulation, aboveground biomass and yield were highest in the 300 kg x hm(-2) treatment. The quadratic model indicated that application rate of 349 to 382 kg N x hm(-2) was optimum for drip-irrigated processing tomatoes in northern Xinjiang.

Use of a digital camera to monitor the growth and nitrogen status of cotton.

The main objective of this study was to develop a nondestructive method for monitoring cotton growth and N status using a digital camera. Digital images were taken of the cotton canopies between emergence and full bloom. The green and red values were extracted from the digital images and then used to calculate canopy cover. The values of canopy cover were closely correlated with the normalized difference vegetation index and the ratio vegetation index and were measured using a GreenSeeker handheld sensor. Models were calibrated to describe the relationship between canopy cover and three growth properties of the cotton crop (i.e., aboveground total N content, LAI, and aboveground biomass). There were close, exponential relationships between canopy cover and three growth properties. And the relationships for estimating cotton aboveground total N content were most precise, the coefficients of determination (R(2)) value was 0.978, and the root mean square error (RMSE) value was 1.479 g m(-2). Moreover, the models were validated in three fields of high-yield cotton. The result indicated that the best relationship between canopy cover and aboveground total N content had an R(2) value of 0.926 and an RMSE value of 1.631 g m(-2). In conclusion, as a near-ground remote assessment tool, digital cameras have good potential for monitoring cotton growth and N status.

Rice photosynthetic productivity and PSII photochemistry under nonflooded irrigation.

Nonflooded irrigation is an important water-saving rice cultivation technology, but little is known on its photosynthetic mechanism. The aims of this work were to investigate photosynthetic characteristics of rice during grain filling stage under three nonflooded irrigation treatments: furrow irrigation with plastic mulching (FIM), furrow irrigation with nonmulching (FIN), and drip irrigation with plastic mulching (DI). Compared with the conventional flooding (CF) treatment, those grown in the nonflooded irrigation treatments showed lower net photosynthetic rate (PN), lower maximum quantum yield (Fv/Fm), and lower effective quantum yield of PSII photochemistry (ΦPSII). And the poor photosynthetic characteristics in the nonflooded irrigation treatments were mainly attributed to the low total nitrogen content (TNC). Under non-flooded irrigation, the PN, Fv/Fm, and ΦPSII significantly decreased with a reduction in the soil water potential, but these parameters were rapidly recovered in the DI and FIM treatments when supplementary irrigation was applied. Moreover, The DI treatment always had higher photosynthetic productivity than the FIM and FIN treatments. Grain yield, matter translocation, and dry matter post-anthesis (DMPA) were the highest in the CF treatment, followed by the DI, FIM, and FIN treatments in turn. In conclusion, increasing nitrogen content in leaf of rice plants could be a key factor to improve photosynthetic capacity in nonflooded irrigation.

Rice performance and water use efficiency under plastic mulching with drip irrigation.

Plastic mulching with drip irrigation is a new water-saving rice cultivation technology, but little is known on its productivity and water-saving capacity. This study aimed to assess the production potential, performance, and water use efficiency (WUE) of rice under plastic mulching with drip irrigation. Field experiments were conducted over 2 years with two rice cultivars under different cultivation systems: conventional flooding (CF), non-flooded irrigation incorporating plastic mulching with furrow irrigation (FIM), non-mulching with furrow irrigation (FIN), and plastic mulching with drip irrigation (DI). Compared with the CF treatment, grain yields were reduced by 31.76-52.19% under the DI treatment, by 57.16-61.02% under the FIM treatment, by 74.40-75.73% under the FIN treatment, which were mainly from source limitation, especially a low dry matter accumulation during post-anthesis, in non-flooded irrigation. WUE was the highest in the DI treatment, being 1.52-2.12 times higher than with the CF treatment, 1.35-1.89 times higher than with the FIM treatment, and 2.37-3.78 times higher than with the FIN treatment. The yield contribution from tillers (YCFTs) was 50.65-62.47% for the CF treatment and 12.07-20.62% for the non-flooded irrigation treatments. These low YCFTs values were attributed to the poor performance in tiller panicles rather than the total tiller number. Under non-flooded irrigation, root length was significantly reduced with more roots distributed in deep soil layers compared with the CF treatment; the DI treatment had more roots in the topsoil layer than the FIM and FIN treatments. The experiment demonstrates that the DI treatment has greater water saving capacity and lower yield and economic benefit gaps than the FIM and FIN treatments compared with the CF treatment, and would therefore be a better water-saving technology in areas of water scarcity.

A transgenic Bm cell line of piggyBac transposon-derived targeting expression of humanized glycoproteins through N-glycosylation.

Glycoproteins have been implicated in a wide variety of important biochemical and biological functions, including protein stability, immune function, enzymatic function, cellular adhesion and others. Unfortunately, there is no therapeutic protein produced in insect system to date, due to the expressed glycoproteins are paucimannosidic N-glycans, rather than the complex, terminally sialylated N-glycans in mammalian cells. In this paper, we cloned the necessary genes in glycosylation of mammalian cells, such as N-acetylglucosaminyltransferase II (Gn-TII), galactosyltransferases (Gal-Ts), 2,6-Sial-T (ST6 GalII)and 2,3-Sial-T (ST3GalIII), and transformed them to silkworm genome of BmN cell line through transgenesis to establish a transgenic Bm cell line of piggyBac transposon-derived targeting expression of humanized glycoproteins. The study supplied a new insect cell line which is practically to produce "bisected" complex N-glycans like in mammalian cells.

Heterologous expression characteristics of Trichoderma viride endoglucanase V in the silkworm, Bombyx mori L.

Efficient degradation of cellulose needs a synergistic reaction of the cellulolytic enzymes, which include exoglucanases, endoglucanases, and ß-1,4-glucosidase. In this study, we used an improved Bac-to-Bac/BmNPV baculovirus expression system, which lacks the virus-encoded chitinase cathepsin (v-cath) genes of Bombyx mori nucleopolyhedrovirus (BmNPV), to express the endoglucanase V (EG V) gene from Trichoderma viride in silkworm BmN cells and silkworm larvae, and analyzed the characteristics of the recombinant enzyme in silkworm larvae. The result showed that an around 36-kDa protein was visualized in BmN cells at 48 h after the second-generation recombinant mBacmid/BmNPV/EG V baculovirus infection. The crude enzyme extract from the recombinant baculoviruses-infected silkworms exhibited a significant maximum activity at the environmental condition of pH 5.0 and a temperature of 50 °C, and increased 39.86% and 37.76% compared with that from blank mBacmid/BmNPV baculovirus-infected silkworms and normal silkworms, respectively. It was stable at pH range from 5.0 to 10.0 and at temperature range from 40 to 60 °C. The availability of large quantities of EG V that the silkworm provides might greatly facilitate the future research and the potential application in industries.

Cloning and characterization of the Bombyx mori ecdysone oxidase.

The physiological titer of molting hormones in insects depends on relative activities of synthesis and degradation pathways. Ecdysone oxidase (EO) is a key enzyme in the inactivation of ecdysteroid. However, there are only a few reports on ecdysteroid inactivation and its enzymes in silkworm. In this study, we cloned and characterized the Bombyx mori EO (BmEO). The BmEO cDNA contains an ORF of 1,695 bp and the deduced protein sequence contains 564 amino acid residues. The deduced protein sequence contains two functional domains of glucose-methanol-choline oxidoreductase in N-terminal and C-terminal. Comparing the expression levels of BmEO in different tissues, high transcription was mainly present in hemocytes. Reduced expression of this enzyme is expected to lead to pathological accumulation of ecdysone in the hemolymph of silkworm larvae or pupae. Our data show that RNA inference of BmEO transcripts resulted in the accumulation of ecdysteroid and death of larvae or pupae. We infer that EO is a crucial element in the physiology of insect development.

Cloning and expression pattern of 3-dehydroecdysone 3ß-reductase (3DE 3ß-reductase) from the silkworm, Bombyx mori L.

Molting in insects is regulated by molting hormones (ecdysteroids), which are also crucial to insect growth, development, reproduction, etc. Ecdysone was inactivated to 3-dehydroecdysone (3DE) under ecdysone oxidase (EO), and followed by NAD(P)H-dependent irreversible reduction to 3-epiecdysteroid under 3DE 3a-reductase. On the other hand, 3-dehydroecdysone undergoes reversible reduction to ecdysone by 3DE 3ß-reductase in the hemolymph. In this article, we cloned and characterized 3-dehydroecdysone 3ß-reductase (3DE 3ß-reductase) in the different tissues and the developing stage from the silkworm, Bombyx mori L. The B. mori 3DE 3ß-reductase cDNA contains an ORF 972 bp and the deduced protein sequence containing 323 amino acid residues. Analysis showed that the deduced 3DE 3ß-reductase belongs to the aldo-keto reductase (AKR) superfamily, which has the NAD(P)-binding domain, indicating that the function of 3DE 3ß-reductase depends on the existence of NAD(P)H. Using Escherichia coli, a high level expression of a fusion polypeptide band of approx. 40 kDa was observed. The high transcription of 3DE 3ß-reductase was mainly observed in the genitalia and fatty bodies in the third day of the fifth-instar larvae, followed next in the head, epidermis, and hemocytes. The expression of 3DE 3ß-reductase in the early of every instar was lower than that in the late of instar. When the titer of 3DE is low, higher expression of 3DE 3ß-reductase is necessary to maintain the ecdysone titer in body through converting 3DE to ecdysone, while the 3DE titer is high, the expression of 3DE 3ß-reductase showed feedback inhibition.

Expression pattern of enzymes related to juvenile hormone metabolism in the silkworm, Bombyx mori L.

The physiological balance of juvenile hormone (JH) in insects depends on its biosynthesis and degradation pathway. Three key enzymes namely, juvenile hormone esterase (JHE), juvenile hormone epoxide hydrolase (JHEH) and juvenile hormone diol kinase (JHDK) are required for degradation in insects. Our present results showed that JHE and JHEH exhibited expression in almost all the tissues. This indicated that JHE and JHEH might degrade JH simultaneously. In addition, the highest levels of JHDK were observed in the midgut, with trace level being found in the malpighian tubule and haemocytes. Since the midgut is a digestive organ and not a JH target, it was hypothesized that both JHE and JHEH hydrolyzed JH to JH diol (JHd) which was then transported to midgut and hydrolyzed further by JHDK, to be finally excreted out of the body. Also the expression studies on JH degradation enzymes in different tissues and stages indicated that the activities of the three enzymes are specific and coincident with the JH functions in silkworm, Bombyx mori L.

Expression of Trichoderma viride endoglucanase III in the larvae of silkworm, Bombyx mori L. and characteristic analysis of the recombinant protein.

Endoglucanase is a part of cellulase which hydrolyzes cellulose into glucose. In this study, we cloned endoglucanase III (EG III) gene from Trichoderma viride strain AS 3.3711 using a PCR-based exon splicing method, and expressed EG III recombinant protein in both silkworm BmN cell line and silkworm larvae with an improved Bac-to-Bac/BmNPV mutant baculovirus expression system, which lacks the chiA and v-cath genes of Bombyx mori nucleopolyhedrovirus (BmNPV). The result showed that around 45 kDa protein was visualized in BmN cells at 48 h after the second generation recombinant mBacmid/BmNPV/EG III baculovirus infection. The enzymes from recombinant baculoviruses infected silkworms exhibited significant maximum enzyme activity at the environmental condition of pH 8.0 and temperature 50°C, and increased 20.94 and 19.13% compared with that from blank mBacmid/BmNPV baculoviruses infected silkworms and normal silkworms, respectively. It was stable at pH range from 5.0 to 9.0 and at temperature range from 40 to 60°C. It provided a possibility to generate transgenic silkworms expressing bio-active cellulase, which can catabolize dietary fibers more efficiently, and it might be of great significance for sericulture industry.

Identification and characterization of two chitin-binding proteins from the peritrophic membrane of the silkworm, Bombyx mori L.

The peritrophic membrane (PM) is a semi-permeable lining of the insect midgut, broadly analogous to the mucous lining of vertebrate gut. The PM proteins are important achievements for the function of the PM. In this study, two chitin-binding proteins (BmPM-P43 and BmPM-P41) from the PM of the silkworm, Bombyx mori, were identified and cloned. These proteins showed the molecular mass of 43 and 41 kDa, respectively. The deduced amino acid sequences codes for a protein of 381 amino acid residues and 364 amino acid residues, containing 12 and 14 cysteine residues followed by similar domain, both of them have 5 cysteine residues in similar position in the C-terminal. The confirmation of these proteins was performed by western blot analysis of recombinant BmPM-P43 and BmPM-P41. The chitin-binding activity analysis showed that the BmPM-P43 and BmPM-P41 could bind to chitin strongly. It is concluded that BmPM-P43 and BmPM-P41 contains a polysaccharide deacetylase domain instead of peritrophin domain, indicated that these two proteins may belong to a new chitin-binding protein family.