Gene signatures of copper metabolism related genes may predict prognosis and immunity status in Ewing's sarcoma.

Background: Cuproptosis is copper-induced cell death. Copper metabolism related genes (CMRGs) were demonstrated that used to assess the prognosis out of tumors. In the study, CMRGs were tested for their effect on TME cell infiltration in Ewing's sarcoma (ES). Methods: The GEO and ICGC databases provided the mRNA expression profiles and clinical features for downloading. In the GSE17674 dataset, 22prognostic-related copper metabolism related genes (PR-CMRGs) was identified by using univariate regression analysis. Subsequently, in order to compare the survival rates of groups with high and low expression of these PR-CMRGs,Kaplan-Meier analysis was implemented. Additionally, correlations among them were examined. The study employed functional enrichment analysis to investigate probable underlying pathways, while GSVA was applied to evaluate enriched pathways in the ES (Expression Set). Through an unsupervised clustering algorithm, samples were classified into two clusters, revealing significant differences in survival rates and levels of immune infiltration. Results: Using Lasso and step regression methods, five genes (TFRC, SORD, SLC11A2, FKBP4, and AANAT) were selected as risk signatures. According to the Kaplan-Meier survival analysis, the high-risk group had considerably lower survival rates than the low-risk group(p=6.013e-09). The area under the curve (AUC) values for the receiver operating characteristic (ROC) curve were 0.876, 0.883, and 0.979 for 1, 3, and 5 years, respectively. The risk model was further validated in additional datasets, namely GSE63155, GSE63156, and the ICGC datasets. To aid in outcome prediction, a nomogram was developed that incorporated risk levels and clinical features. This nomogram's performance was effectively validated through calibration curves.Additionally, the study evaluated the variations in immune infiltration across different risk groups, as well as high-expression and low-expression groups. Importantly, several drugs were identified that displayed sensitivity, offering potential therapeutic options for ES. Conclusion: The findings above strongly indicate that CMRGs play crucial roles in predicting prognosis and immune status in ES.

A nomogram model of spectral CT quantitative parameters and clinical characteristics predicting lymphovascular invasion of gastric cancer.

Objective: The study established a nomogram based on quantitative parameters of spectral computed tomography (CT) and clinical characteristics, aiming to evaluate its predictive value for preoperative lymphovascular invasion (LVI) in gastric cancer (GC). Methods: From December 2019 to December 2021, 171 patients with pathologically confirmed GC were retrospectively collected with corresponding clinical data and spectral CT quantitative data. Patients were divided into LVI-positive and LVI-negative groups based on their pathological results. The univariate and multivariate logistic regression analyses were used to identify the risk factors and construct a nomogram. The calibration curve and receiver operating characteristic (ROC) curve were adopted to evaluate the predictive accuracy of nomogram. Results: Four clinical characteristics or spectral CT quantitative parameters, including Borrmann classification (P = 0.039), CA724 (P = 0.007), tumor thickness (P = 0.031), and iodine concentration in the venous phase (VIC) (P = 0.004) were identified as independent factors for LVI in GC patients. The nomogram was established based on the four factors, which had a potent predictive accuracy in the training, internal validation and external validation cohorts, with the area under the ROC curve (AUC) of 0.864 (95% CI, 0.798-0.930), 0.964 (95% CI, 0.903-1.000) and 0.877 (95% CI, 0.759-0.996), respectively. Conclusion: This study constructed a comprehensive nomogram consisting spectral CT quantitative parameters and clinical characteristics of GC, which exhibited a robust efficiency in predicting LVI in GC patients.

Research on machine vision and deep learning based recognition of cotton seedling aphid infestation level.

Aphis gossypii Glover is a major insect pest in cotton production, which can cause yield reduction in severe cases. In this paper, we proposed the A. gossypii infestation monitoring method, which identifies the infestation level of A. gossypii at the cotton seedling stage, and can improve the efficiency of early warning and forecasting of A. gossypii, and achieve precise prevention and cure according to the predicted infestation level. We used smartphones to collect A. gossypii infestation images and compiled an infestation image data set. And then constructed, trained, and tested three different A. gossypii infestation recognition models based on Faster Region-based Convolutional Neural Network (R-CNN), You Only Look Once (YOLO)v5 and single-shot detector (SSD) models. The results showed that the YOLOv5 model had the highest mean average precision (mAP) value (95.7%) and frames per second (FPS) value (61.73) for the same conditions. In studying the influence of different image resolutions on the performance of the YOLOv5 model, we found that YOLOv5s performed better than YOLOv5x in terms of overall performance, with the best performance at an image resolution of 640×640 (mAP of 96.8%, FPS of 71.43). And the comparison with the latest YOLOv8s showed that the YOLOv5s performed better than the YOLOv8s. Finally, the trained model was deployed to the Android mobile, and the results showed that mobile-side detection was the best when the image resolution was 256×256, with an accuracy of 81.0% and FPS of 6.98. The real-time recognition system established in this study can provide technical support for infestation forecasting and precise prevention of A. gossypii.

Does Intraoperative Multi-Drug Wound Infiltration Relieve Postoperative Pain Following Single-Level TLIF Surgery? A Randomized Controlled Trial.

BACKGROUND: How to minimize postoperative pain following spinal surgery has been a great challenge for both surgeons and patients. We hypothesized that intraoperative multi-drug wound infiltration could relieve postoperative pain following single-level transforaminal lumbar interbody fusion (TLIF) surgery. OBJECTIVES: To evaluate the effect of intraoperative multi-drug wound infiltration for postoperative pain following single-level TLIF surgery. STUDY DESIGN: A randomized, double-blinded controlled trial (RCT). SETTING: Department of Orthopaedic Surgery, Qilu Hospital of Shandong University. METHODS: The RCT enrolled 50 patients with 25 cases in 2 groups. The study group received intraoperative wound infiltration of mixed solution with lidocaine, ropivacaine, and epinephrine before wound closure. The control group was infiltrated with an equal amount of normal saline. The primary outcome measure was the visual analog scale (VAS) of postoperative incision pain. The secondary outcome measures were the postoperative opioids dosage, the time of first analgesic demand, and the Oswestry Disability Index (ODI). RESULTS: The VAS of postoperative pain in the study group was significantly lower than the control group within postoperative 24 hours. The opioid dosage was significantly less and the time of first analgesic demand of patient-controlled analgesia (PCA) in the study group was significantly longer than the control group. None of the patients in the study group required analgesic supplementation. The side effects of opioids were significantly less in the study group. There was no significant difference in ODI, operation time, intraoperative blood loss, postoperative drainage, and postoperative incision complications between the 2 groups. LIMITATIONS: Single-center study for single-level TLIF surgery. CONCLUSIONS: Intraoperative multi-drug wound infiltration before closure could significantly relieve postoperative pain following single-level TLIF surgery.

Comparison of Clinical Outcomes and Muscle Invasiveness between Unilateral Biportal Endoscopic Discectomy and Percutaneous Endoscopic Interlaminar Discectomy for Lumbar Disc Herniation at L5/S1 Level.

OBJECTIVE: Both unilateral biportal endoscopic discectomy (UBED) and percutaneous endoscopic interlaminar discectomy (PEID) could achieve favorable outcomes for lumbar disc herniation (LDH). There are limited studies comparing the two different methods of endoscopic discectomy. The objective was to comprehensively compare the clinical outcome and muscle invasiveness of UBED and PEID for the treatment of LDH at L5/S1 level with at least 1-year follow-up. METHODS: The retrospective cohort study enrolled 106 LDH patients of L5/S1 level from January 2018 to December 2020. There were 51 patients who underwent UBED (22 males and 29 females, 43.8 ± 14.2 years old) and 55 patients underwent PEID (28 males and 27 females, 42.3 ± 13.8 years old). Clinical outcomes and surgical invasiveness were compared between the two groups for at least 1 year follow-up. Clinical outcomes included visual analogue scale (VAS) scores, Oswestry Disability Index (ODI), complications, recurrence of LDH, intraoperative anesthesia time, operative time, number of intraoperative fluoroscopies, and postoperative length of stay. Surgical invasiveness was evaluated with serum CPK level and change rate of lean multifidus cross-sectional area (LMCSA). Independent-sample t test and paired sample t test were used to compare continuous data. Chi-square test and Fisher's precision probability tests were used to analyze the categorical data. RESULTS: Both groups achieved favorable clinical outcomes at the last follow-up, including VAS and ODI (all Ps <0.05). The intraoperative anesthesia time for UBED was longer, but with no difference of operative time. As for intraoperative fluoroscopy times (2.5 vs 2.4), postoperative length of stay (2.1 vs 2.0 days), postoperative complications (5.9% vs 3.6%), there were also no significant difference. The serum CPK level and change rate of LMCSA for UBED was higher than PEID at postoperative 1st day. At the last follow-up, there was no significant difference in the change rate of LMCSA between the two groups (P = 0.096). CONCLUSIONS: Both UBED and PEID could achieve favorable clinical outcomes for the treatment of L5/S1 LDH. Despite UBED is more invasive, the radiological manifestation of paraspinal muscle invasiveness was equal at last follow-up with at least 1 year. UBED is a safe and innovative alternative choice for treatment of LDH at L5/S1 level.

Electrostatic Interactions Are the Primary Determinant of the Binding Affinity of SARS-CoV-2 Spike RBD to ACE2: A Computational Case Study of Omicron Variants.

To explore the mechanistic origin that determines the binding affinity of SARS-CoV-2 spike receptor binding domain (RBD) to human angiotensin converting enzyme 2 (ACE2), we constructed the homology models of RBD-ACE2 complexes of four Omicron subvariants (BA.1, BA.2, BA.3 and BA.4/5), and compared them with wild type complex (RBDWT-ACE2) in terms of various structural dynamic properties by molecular dynamics (MD) simulations and binding free energy (BFE) calculations. The results of MD simulations suggest that the RBDs of all the Omicron subvariants (RBDOMIs) feature increased global structural fluctuations when compared with RBDWT. Detailed comparison of BFE components reveals that the enhanced electrostatic attractive interactions are the main determinant of the higher ACE2-binding affinity of RBDOMIs than RBDWT, while the weakened electrostatic attractive interactions determine RBD of BA.4/5 subvariant (RBDBA.4/5) lowest ACE2-binding affinity among all Omicron subvariants. The per-residue BFE decompositions and the hydrogen bond (HB) networks analyses indicate that the enhanced electrostatic attractive interactions are mainly through gain/loss of the positively/negatively charged residues, and the formation or destruction of the interfacial HBs and salt bridges can also largely affect the ACE2-binding affinity of RBD. It is worth pointing out that since Q493R plays the most important positive contribution in enhancing binding affinity, the absence of this mutation in RBDBA.4/5 results in a significantly weaker binding affinity to ACE2 than other Omicron subvariants. Our results provide insight into the role of electrostatic interactions in determining of the binding affinity of SARS-CoV-2 RBD to human ACE2.

Greater flood risks in response to slowdown of tropical cyclones over the coast of China.

The total amount of rainfall associated with tropical cyclones (TCs) over a given region is proportional to rainfall intensity and the inverse of TC translation speed. Although the contributions of increase in rainfall intensity to larger total rainfall amounts have been extensively examined, observational evidence on impacts of the recently reported but still debated long-term slowdown of TCs on local total rainfall amounts is limited. Here, we find that both observations and the multimodel ensemble of Global Climate Model simulations show a significant slowdown of TCs (11% in observations and 10% in simulations, respectively) from 1961 to 2017 over the coast of China. Our analyses of long-term observations find a significant increase in the 90th percentile of TC-induced local rainfall totals and significant inverse relationships between TC translation speeds and local rainfall totals over the study period. The study also shows that TCs with lower translation speed and higher rainfall totals occurred more frequently after 1990 in the Pearl River Delta in southern China. Our probability analysis indicates that slow-moving TCs are more likely to generate heavy rainfall of higher total amounts than fast-moving TCs. Our findings suggest that slowdown of TCs tends to elevate local rainfall totals and thus impose greater flood risks at the regional scale.

Intranasal Application of Budesonide Attenuates Lipopolysaccharide-Induced Acute Lung Injury by Suppressing Nucleotide-Binding Oligomerization Domain-Like Receptor Family, Pyrin Domain-Containing 3 Inflammasome Activation in Mice.

AIM: To investigate the protective effects of budesonide against lipopolysaccharide- (LPS-) induced acute lung injury (ALI) in a murine model and its underlying mechanism. METHODS: Adult male C57BL/6 mice were divided into three groups: control, ALI, and ALI + budesonide groups. LPS (5 mg/kg) was intratracheally injected to induce ALI in mice. Budesonide (0.5 mg/kg) was intranasally given 1 h before LPS administration in the ALI + budesonide group. Twelve hours after LPS administration, all mice were sacrificed. Hematoxylin-eosin staining and pathological scores were used to evaluate pathological injury. Bronchoalveolar lavage was performed. The numbers of total cells, neutrophils, and macrophages in the bronchoalveolar lavage fluid (BALF) were counted. Enzyme-linked immunosorbent assay was employed to detect the proinflammatory cytokines in BALF and serum, including tumor necrosis factor- (TNF-) α, monocyte chemoattractant protein- (MCP-) 1, and interleukin- (IL-) 1ß. The expression of the nucleotide-binding oligomerization domain-like receptor family, pyrin domain-containing 3 (NLRP3) inflammasome was detected by western blotting. A lethal dose of LPS (40 mg/kg, intraperitoneally) was injected to evaluate the effects of budesonide on survival rates. RESULTS: Budesonide pretreatment dramatically attenuated pathological injury and reduced pathological scores in mice with ALI. Budesonide pretreatment obviously reduced the numbers of total cells, neutrophils, and macrophages in the BALF of mice with ALI. Additionally, budesonide dramatically reduced TNF-α and MCP-1 expression in the BALF and serum of mice with ALI. Budesonide significantly suppressed NLRP3 and pro-caspase-1 expression in the lung and reduced IL-1ß content in the BALF, indicating that budesonide inhibited the activation of the NLRP3 inflammasome. Furthermore, we found that budesonide improved the survival rates of mice with ALI receiving a lethal dose of LPS. CONCLUSION: Suppression of NLRP3 inflammasome activation in mice via budesonide attenuated lung injury induced by LPS in mice with ALI.

Resistance of alfalfa and Indian mustard to Cd and the correlation of plant Cd uptake and soil Cd form.

Phytoremediation of heavy metal-contaminated soil is considered to be one ecological environmental protection way that is effective and economical. The selection of suitable hyperaccumulators is a key issue for phytoremediation of heavy metal-contaminated soil. Pot experiments were conducted to study the effects of different Cd levels (0, 75, 150, 300, and 600 mg kg-1 Cd) on the dry weight, antioxidant enzyme activities, malondialdehyde (MDA) contents, Cd concentration, Cd accumulation, and soil Cd form distribution ratio (FDC) of alfalfa (Medicago sativa L.) and Indian mustard (Brassica juncea L.). The correlations between Cd concentration in shoots and roots of alfalfa and Indian mustard and soil Cd form were also investigated. The results showed that with the increase of soil Cd levels, dry weight of shoot and root of alfalfa and Indian mustard significantly decreased, which decreased by 50.0-71.8% and 29.6-59.3% (alfalfa), 59.6-89.0% and 64.3-74.8% (India mustard), respectively, compared with the control. With the increase of soil Cd level, superoxide dismutase (SOD) activity in shoot of alfalfa significantly increased. Catalase (CAT) activity and malondialdehyde contents in shoots and roots of alfalfa and Indian mustard, as well as superoxide dismutase activity in the roots of alfalfa and Indian mustard increased first and then decreased with the increase of soil Cd level. With increasing Cd stress, Cd concentration in shoots and roots of alfalfa and Indian mustard significantly increased. At soil Cd level of 75 mg kg-1, Cd concentrations in shoots of alfalfa and Indian mustard exceeded the critical value of Cd-hyperaccumulator (100 mg kg-1), which was 356.46 mg kg-1 and 308.74 mg kg-1, respectively. Cadmium concentrations in shoots and roots of plants were in the order of that of alfalfa > Indian mustard; total Cd accumulation in the aboveground tissues and roots of the plants was in the order of that of Indian mustard > alfalfa at the same Cd level. With increasing soil Cd level, Cd concentrations of exchangeable form (EXC-Cd), carbonate-bound form (CAB-Cd), iron-manganese oxide-bound form (FeMn-Cd), organic-bound form (OM-Cd), and residual form (RES-Cd) showed an increasing trend. The form distribution ratio of soil Cd forms in alfalfa's rhizosphere was in the order of that of exchangeable form Cd > carbonate-bound form Cd > iron-manganese oxide-bound form Cd > residual form Cd > organic-bound form Cd. Except for organic-bound form Cd, soil Cd forms were significantly positively correlated with Cd concentration in shoot and root (P < 0.01). Comprehensively considering the biomass and Cd accumulation, Indian mustard is more suitable as remediation material for soil Cd pollution.

B7-H3 expression and its correlation with clinicopathologic features, angiogenesis, and prognosis in intrahepatic cholangiocarcinoma.

This study was designed to explore the expression of B7-H3 in human intrahepatic cholangiocarcinoma (ICC) and its association with the clinicopathologic factors. In the current study, the expression of B7-H3 in 45 patients with intrahepatic cholangiocarcinoma and 8 patients with hepatolithiasis was analyzed by immunohistochemistry, which revealed that B7-H3 was not expressed in hepatolithiatic tissues, but positively expressed in 57.8% (26/45) of the ICC cases. The expression of B7-H3 was significantly associated with lymph node metastases and venous invasion. A positive correlation was also observed between the expression of B7-H3 and MVD, an index for tumor angiogenesis. Further survival analysis indicated that patients with B7-H3 negative expression had higher overall survival (OS) and cancer-specific survival (CSS) rates than those with B7-H3 positive expression. Multivariate analysis revealed that B7-H3 expression was an independent prognostic indicator for poor OS and CSS of ICC patients. Our results suggest that B7-H3 may be a valuable biomarker in determining tumor progression and prognosis of intrahepatic cholangiocarcinoma. It is also a potential target for antivascular therapy of ICC.

[Effect of Nano Zeolite on Growth, Activity of Antioxidant Enzyme, and Chemical Fractions and Concentration of Cd in Chinese Cabbage].

Pot experiment was carried out to investigate the influence of nano zeolite (NZ) and ordinary zeolite (OZ) on the growth, activities of antioxidant enzyme, Cd fraction and Cd concentration in two varieties of Chinese cabbage. The results showed that the activities of SOD, CAT and POD in the shoot and root of two varieties increased, and the dry weight of shoot and the total dry weight of Chinese cabbage increased by 4.5%-96.5% and 3.4%-88.4%, respectively. The application of zeolite effectively reduced the Cd concentration of shoot and root in the two varieties of Chinese cabbage, and the reduction range increased with the increase of the application amount of zeolite. Under the low Cd condition (1mg·kg-1 Cd), the Cd concentration of edible parts (shoots) in SD4 variety and XJC3 variety decreased by 1.0%-75.0% and 19.5%-68.9% (except the Cd1+OZ5/OZ10 processing), respectively. Under the high Cd condition (5 mg·kg-1 Cd), the Cd concentration of the edible parts (shoots) in SD4 variety and XJC3 variety decreased by 7.2%-53.2%(except Cd5+OZ10 processing) and 0.7%-63.0%, respectively. The Cd concentrations of the edible parts (shoots) in tow varieties of Chinese cabbage in the nano zeolite treatments decreased by 10.5%-65.7% compared with the ordinary zeolite treatments. In Cd contaminated soils (1 mg·kg-1 and 5 mg·kg-1 Cd),) the total extraction amount of Cd in the edible parts (shoots) of SD4 variety reduced by 12.4%-68.8% and 13.2%-55.6% (except of Cd5+OZ5 processing), and the total extraction amount of Cd in the edible parts (shoots) of XJC3 variety reduced by 9.4%-71.5% and 3.1%-38.7% (except of Cd1+OZ5 processing), respectively. The application amount of nano zeolite (NZ) and ordinary zeolite (OZ) was more, the reduction range of the total extraction amount of Cd in the edible parts (shoots) of two varieties of Chinese cabbage was greater.

Isolation of prostrate turfgrass mutants via screening of dwarf phenotype and characterization of a perennial ryegrass prostrate mutant.

Prostrate turf varieties are desirable because of their increased low mowing tolerance, heat resistance, traffic resistance and ground coverage compared with upright varieties. Mutation breeding may provide a powerful tool to create prostrate varieties, but there are no simple, straightforward methods to screen for such mutants. Elucidation of the molecular basis of the major 'green revolution' traits, dwarfism and semi-dwarfism, guided us to design a simple strategy for isolating dwarf mutants of perennial ryegrass (Lolium perenne L.). We have shown that gamma-ray-mediated dominant dwarf mutants can be easily screened for at the three-leaf stage. About 10% of dwarf mutant lines also displayed a prostrate phenotype at mature stages (>10 tillers). One prostrate line, Lowboy I, has been characterized in detail. Lowboy I had significantly shorter canopy, leaf blade and internode lengths compared with wild type. Lowboy I also exhibited greater tolerance to low mowing stress than wild type. Exogenous gibberellic acid (GA) restored Lowboy I to a wild-type phenotype, indicating that the dwarf and prostrate phenotypes were both due to GA deficiency. We further showed that phenotypes of Lowboy I were dominant and stably inherited through sexual reproduction. Prostrate turfgrass mutants are difficult to screen for because the phenotype is not observed at young seedling stages, therefore our method represents a simple strategy for easily isolating prostrate mutants. Furthermore, Lowboy I may provide an outstanding germplasm for breeding novel prostrate perennial ryegrass cultivars.

[Effect of Nano Zeolite on Chemical Fractions of Cd in Soil and Uptake by Chinese Cabbage at Different Soil pH and Cadmium Levels].

Incubation experiments were carried out to investigate the influence of different nano zeolite(NZ) and ordinary zeolite(OZ) levels(0, 5, 10 and 20 g·kg-1) on the fraction distribution coefficient (FDC) of Cd and soil CEC at different soil pH (4, 5, 6, 7 and 8) when exposed to different cadmium(Cd) levels(1, 5, 10 and 15 mg·kg-1), and pot experiment were carried out to investigate the effects of nano zeolite(NZ) and ordinary zeolite(OZ) on the growth, Cd concentration and Cd accumulation of Chinese cabbage. The results showed that nano zeolite and ordinary zeolite decreased the concentration and FDC of exchangeable Cd (EX-F), and increased the concentration and FDC of carbonate(CAB-F), Fe-Mn oxide(FMO-F), organic matter (OM-F) and residual fraction(RES-F) in incubation experiments. At the end of incubation, the FDC of soil exchangeable Cd decreased from 72.0%-88.0% to 2.4%-10.7%. The decreasing effect of zeolite on the concentration and FDC of exchangeable Cd (EX-F) increased with the increase of zeolite, and the decreasing effect of nano zeolite (NZ) was better than that of ordinary zeolite (OZ). During the culture of 28 d, the concentration of different Cd fractions in soil was in order of EX-F>RES-F>FMO-F>CAB-F>OM-F under different pH conditions. Exchangeable fraction Cd was the dominant fraction of Cd in soil during the whole incubation. Soil CEC had significant negative correlation with soil exchangeable Cd (EX-F) (P<0.01), and significant positive correlation with the concentrations of Fe-Mn oxide(FMO-F) and organic matter (OM-F) in soil(P<0.01). Nano zeolite and ordinary zeolite effectively increased soil CEC, and soil CEC increased with the increase of the pH value of soil in the zeolite treatments. Significant negative correlation was found in soil pH with soil exchangeable Cd (EX-F)(P<0.01). The dry weight of plant tissues in Chinese cabbage increased by 14.3%-131.4% in the presence of nano zeolite(NZ) and ordinary zeolite(OZ), and Cd concentration of shoot and root decreased by 1.0%-75.0% and 3.8%-53.2%, respectively. Higher concentration and accumulation of Cd were observed in XJC3 variety than those in SD 4 variety. Compared with ordinary zeolite (OZ), nano zeolite (NZ) was better in increasing the biomass of Chinese cabbage as well as decreasing accumulation of Cd in Chinese cabbage.

[Effect of Nano Zeolite on Chemical Fractions of Cd in Soil and Its Uptake by Cabbage].

Incubation experiments were carried out to investigate the influence of different nano zeolite (NZ) and ordinary zeolite (OZ) levels(0, 5, 10 and 20 g · kg⁻¹) on the change trends in fraction distribution coefficient (FDC) of Cd when exposed to different Cadmium (Cd) levels (1, 5, 10 and 15 mg · kg⁻¹), and pot experiments were carried out to investigate their influence on soil Cd fraction and Cd uptake by cabbage. The results in incubation experiments showed that the application of nano zeolite as well as ordinary zeolite effectively decreased the FDC of exchangeable Cd and increased the FDC of Fe-Mn oxide fraction. The FDC of soil Cd from 0 d to 28 d was deceased at first, then increased and tended to be stable, and finally increased. At the end of incubation, the FDC of soil exchangeable Cd decreased from 72.0%-88.0% to 30.0%-66.4%. Exchangeable fraction Cd was the most dominant Cd fraction in soil during the whole incubation. The results in pot experiment indicated that the application of nano zeolite and ordinary zeolite decreased the concentration and FDC of soil exchangeable Cd, and concurrently the concentration and FDC of Cd in carbonate, Fe-Mn oxide, organic matter and residual fraction were increased. The lowest EX-Cd was observed in the treatment with high dose of nano zeolite (20 g · kg⁻¹). The FDC of exchangeable Cd showed significant negative relationship with the soil pH (P < 0.05), and was concurrently extremely positively correlated with Cd concentration in shoot and root of cabbage (P < 0.01). Soil pH increased by 1.8%-45.5% and 6.1%-54.3% in the presence of zeolite when exposed to 5 mg · kg⁻¹ 1 and Cd, respectively; FDC of exchangeable Cd decreased by 16.3%-47.7% and 16.2%-46.7%; Cd concentration in each tissues of cabbage decreased by 1.0%-75.0% and 3.8%-53.2%, respectively. Moreover, the reduction effect of nano zeolite on soil and plant Cd was better than that of ordinary zeolite. The growth of cabbage was stimulated by low and medium zeolite doses (≤ 10 g · kg⁻¹), while inhibited by high zeolite doses (20 g · kg⁻¹). Compared to ordinary zeolite, the biomass of Chinese cabbage was significantly increased by Nano zeolite, while the exchangeable Cd in soil as well as Cd concentration and Cd accumulation of cabbage were significantly reduced.

[Effect of Ryegrass and Arbuscular Mycorrhizal on Cd Absorption by Varieties of Tomatoes and Cadmium Forms in Soil].

Field trial was carried out to investigate the effects of ryegrass and arbuscular mycorrhizal single or compound treatment to two varieties of tomato ("Defu mm-8" and "Luobeiqi") on the plant growth, concentrations and accumulations of Cd as well as the impact on microorganisms, enzyme activities, pH and Cd forms in soil when exposed to Cd (5.943 mg · kg⁻¹). The results showed that dry weights of fruit, root, stem, leaf and plant significantly increased by single or compound treatment of ryegrass and arbuscular mycorrhizal by 14.1%-38.4% and 4.2%-18.3%, 20.9%-31.5% and 8.4%-10.3%, 13.0%-16.8% and 3.0%-9.5%, 10.7%- 16.8% and 2.7%-7.6%, 14.3%-36.6% and 4.5%-16.8%, respectively. The amounts of bacteria, fungi, actinomycetes of soil and the activities of urease, invertase, acid phosphatase, catalase in soil were increased by single or compound treatment of ryegrass and arbuscular mycorrhizal, and the soil microorganism amounts and enzyme activities significantly differed between the two varieties of tomato and treatments (P < 0.05). Soil pH was increased by single or compound treatment of ryegrass and arbuscular mycorrhizal, while the concentrations of EXC-Cd, CAB-Cd, Fe-Mn-Cd and total Cd in soil were decreased, and the total Cd content was decreased by 16.9%-27.8%. Cadmium concentrations in fruit, leaf, stem and root of both varieties were significantly decreased by 6.9%-40.9%, 5.7%-40.1%, 4.6%-34.7% and 9.8%-42.4%, respectively. Cadmium accumulations in tomato were in order of leaf > stem > root > fruit. Comparing the two tomato varieties, Cd concentrations and Cd accumulations in fruit and plant were in order of "Luobeiqi" < "Defu mm-8" in the presence or absence of single or compound treatment of ryegrass and arbuscular mycorrhizal.

Molecular cloning and differential expression analysis of a squalene synthase gene from Dioscorea zingiberensis, an important pharmaceutical plant.

Diosgenin is a steroid derived from cholesterol in plants and used as a typical initial intermediate for synthesis of numerous steroidal drugs in the world. Commercially, this compound is extracted mainly from the rhizomes or tubers of some Dioscorea species. Squalene synthase (SQS: EC 2.5.1.21) catalyzes the condensation of two molecules of farnesyl diphosphate to form squalene, the first committed step for biosynthesis of plant sterols including cholesterol, and is thought to play an important role in diosgenin biosynthesis. A full-length cDNA of a putative squalene synthase gene was cloned from D. zingiberensis and designated as DzSQS (Genbank Accession Number KC960673). DzSQS was contained an open reading frame of 1,230 bp encoding a polypeptide of 409 amino acids with a predicted molecular weight of 46 kDa and an isoelectric point of 6.2. The deduced amino acid sequence of DzSQS shared over 70 % sequence identity with those of SQSs from other plants. The truncated DzSQS in which 24 amino acids were deleted from the carboxy terminus was expressed in Escherichia coli, and the resultant bacterial crude extract was incubated with farnesyl diphosphate and NADPH. GC-MS analysis showed that squalene was detected in the in vitro reaction mixture. Quantitative real-time PCR analysis revealed that DzSQS was expressed from highest to lowest order in mature leaves, newly-formed rhizomes, young leaves, young stems, and two-year-old rhizomes of D. zingiberensis.

Improvement of amorpha-4,11-diene production by a yeast-conform variant of Vitreoscilla hemoglobin.

Amorpha-4,11-diene is the precursor of the antimalarial compound artemisinin. The effect of Vitreoscilla hemoglobin (VHb) and its yeast-conform variant (VHbm) on amorpha-4,11-diene production in engineered Saccharomyces cerevisiae was investigated. First, the VHb gene was mutated to the yeast-conform variant VHbm based on step-by-step extension of a short region of the gene through a series of polymerase chain reactions (PCR). The artificial VHbm gene contained codons preferred by the yeast translation machinery. Two yeast expression vectors containing VHb or VHbm gene were constructed and introduced into the amorpha-4,11-diene-producing strain S. cerevisiae WK1 to form WK1[VHb] and WK1[VHbm], respectively. Western blot and CO-difference spectrum absorbance assay showed that VHb and VHbm were successfully expressed. In shake flasks, VHbm expression conferred higher cell growth than VHb expression. GC-MS results indicated the amorpha-4,11-diene production in WK1[VHbm] and WK1[VHb] was 3- and 2-fold higher than that in WK1, respectively. This suggests that VHb might improve the amorpha-4,11-diene production in engineered S. cerevisiae.

[Intragastric and subcutaneous administration of yolk antibodies against adipocyte plasma membrane proteins on growth and fat deposition in rats].

OBJECTIVE: To study the effects of different administrations of antibodies against adipocyte plasma membrane proteins on growth and fat deposition in rats. METHOD: Ninety six female SD rats that weighed approximately 140 g were allotted randomly into four groups which were given negitive control yolk and positive yolk containing antibody (IgY) against adipocyte plasma membrane (APM) proteins by intragastric administration (i.g.) and subcutaneous injection (s.c.) respectively. Rats were given 1 ml of yolk for every three days in i.g. groups. Rats were given 1 ml of yolk for 4 consecutive days, and the procedure was repeated after one month. The trial lasted for 75 days after which rats were slaughtered for carcass analysis and sampling. RESULTS: The body weight gain and food intake of rats were not different between treatment and control groups. In i.g. goups, positive yolk decreased mesemteric fat index, paramertrial fat index and perirenal fat index (P < 0.05), reduced serum triglycerides (P < 0.05) and increased serum free fatty acids (FFA) (P < 0.01), and also decreased serum leptin, insulin and TNF-alpha levels ( P < 0.01 or P < 0.05), but did not affect gastrocnemius muscle index and serum cholesterol. In s.c. groups, positive yolk increased gastrocnemius muscle index (P < 0.05), reduced serum triglycerides and serum leptin (P < 0.01), increased serum TNF-alpha (P < 0.05), but did not affect adipose tissue depots, serum FFA, cholesterol and insulin. CONCLUSION: Administration of yolk antibody against APM proteins could effectively improve body composition of rats, and the treatment by intragastric administration could give better effect than by subcutaneous injection.

Molecular cloning and characterization of an acetylcholinesterase cDNA in the brown planthopper, Nilaparvata lugens.

A full cDNA encoding an acetylcholinesterase (AChE, EC 3.1.1.7) was cloned and characterized from the brown planthopper, Nilaparvata lugens Stål (Hemiptera: Delphacidae). The complete cDNA (2467 bp) contains a 1938-bp open reading frame encoding 646 amino acid residues. The amino acid sequence of the AChE deduced from the cDNA consists of 30 residues for a putative signal peptide and 616 residues for the mature protein with a predicted molecular weight of 69,418. The three residues (Ser242, Glu371, and His485) that putatively form the catalytic triad and the six Cys that form intra-subunit disulfide bonds are completely conserved, and 10 out of the 14 aromatic residues lining the active site gorge of the AChE are also conserved. Northern blot analysis of poly(A)+ RNA showed an approximately 2.6-kb transcript, and Southern blot analysis revealed there likely was just a single copy of this gene in N. lugens. The deduced protein sequence is most similar to AChE of Nephotettix cincticeps with 83% amino acid identity. Phylogenetic analysis constructed with 45 AChEs from 30 species showed that the deduced N. lugens AChE formed a cluster with the other 8 insect AChE2s. Additionally, the hypervariable region and amino acids specific to insect AChE2 also existed in the AChE of N. lugens. The results revealed that the AChE cDNA cloned in this work belongs to insect AChE2 subgroup, which is orthologous to Drosophila AChE. Comparison of the AChEs between the susceptible and resistant strains revealed a point mutation, Gly185Ser, is likely responsible for the insensitivity of the AChE to methamidopho in the resistant strain.