Bilirubin influences the predictive effect of body mass index on hospital mortality in critically ill patients.

Introduction: Body mass index (BMI) can predict mortality in critically ill patients. Moreover, mortality is related to increased bilirubin levels. Thus, herein, we aimed to investigate the effect of bilirubin levels on the usefulness of BMI in predicting mortality in critically ill patients. Methods: Data were extracted from the Medical Information Mart for Intensive Care (MIMIC IV) database. Patients were divided into two groups according to their total bilirubin levels within 24 h. Cox proportional hazard regression models were applied to obtain adjusted hazard ratios and 95 % confidence intervals for the correlation between BMI categories and hospital mortality. The dose-response relationship was flexibly modeled using a restricted cubic spline (RCS) with three knots. Results: Of the 14376 patients included, 3.4 % were underweight, 29.3 % were of normal body weight, 32.2 % were overweight, and 35.1 % were obese. For patients with total bilirubin levels <2 mg/dL, hospital mortality was significantly lower in patients with obesity than in normal body weight patients (p < 0.05). However, the opposite results were observed for patients with total bilirubin levels ≥2 mg/dL. The Cox proportional hazard regression models suggested that the risk of death was lower in patients with overweightness and obesity than in normal body weight patients when the total bilirubin levels were <2 mg/dL, but not in the other case (total bilirubin levels ≥2 mg/dL). RCS analyses showed that, for patients with total bilirubin levels <2 mg/dL, the risk of death gradually decreased with increasing BMI. Conversely, for patients with total bilirubin levels ≥2 mg/dL, this risk did not decrease with increasing BMI until reaching obesity, after which it increased rapidly. Conclusion: BMI predicted the risk of death differently in critically ill patients with different bilirubin levels.

Pathogenic mechanism of Eimeria tenella autophagy activation of chicken embryo cecal epithelial cells induced by Eimeria tenella.

Eimeria tenella mainly invades and develops into cecal epithelial cells of chickens, resulting in cecal epithelial cell damage. Infectious intracellular pathogens possibly act by influencing the autophagy process after invading cells. The interaction between E. tenella and the autophagy of host cells was explored by infecting E. tenella with chick embryo cecal epithelial cells. Transmission electron microscopy, laser confocal microscopy, and Western blot analysis were used to demonstrate that E. tenella infection could induce autophagy in host cells. Results showed that infection with E. tenella induced the formation of autophagosomes in cells. The expression of ATG 5, Beclin-1, and LC3B-II proteins were significantly (P < 0.01) increased after E. tenella infected host cells. Expression of p62 protein levels were significantly (P < 0.01) decreased in host cells infected with E. tenella. Chloroquine (CQ) significantly (P < 0.01) increased the expression levels of LC3B-II and P62 in E. tenella-infected host cells. Rapamycin (RAPA) induced autophagy in host cells, thus reducing the intracellular infection of E. tenella. By contrast, the infection rate of E. tenella increased in cells treated with 3-Methyladenine (3-MA). Hence, E. tenella sporozoite infection could induce autophagy activation in chick embryo cecal epithelial cells, and enhanced autophagy could reduce the infection rate of E. tenella.

Role of EtMIC4 EGF-like in regulating the apoptosis of Eimeria tenella host cells via the EGFR pathway.

This study aimed to explore the role and key point of EtMIC4 EGF-like recombinant protein in regulating the apoptosis of Eimeria tenella host cells via the epidermal growth factor receptor (EGFR) pathway. The cells were treated with EtMIC4 EGF-like protein, EGFR-specific siRNA, or both. Infection and apoptosis rates as well as dynamic changes in the key genes and proteins of the EGFR signaling pathway in the host cells were determined. Results showed that the E. tenella and EtMIC4 EGF-like group had the highest infection rate (P < 0.01). In cells treated with EtMIC4 EGF-like for 4 to 24 h, the apoptosis rate was significantly decreased (P < 0.01) and the relative mRNA expression and protein phosphorylation levels of EGFR, protein kinase B (AKT), and extracellular regulated protein kinases (ERK) were significantly increased (P < 0.01). In E. tenella sporozoites infected for 4 to 96 h, the rate of host cell apoptosis induced by E. tenella infection was significantly (P < 0.01) reduced by EtMIC4 EGF-like. The relative mRNA expression and protein phosphorylation levels of EGFR, AKT, and ERK in the host cells of E. tenella + EtMIC4 EGF-like group were significantly increased (P < 0.01). These results indicated that E. tenella could activate the EGFR pathway through EtMIC4 EGF-like and regulate the expression of key genes in the AKT and ERK signaling pathways, thereby inhibiting cell apoptosis.

The role of Ca2+ in the injury of host cells during the schizogenic stage of E. tenella.

Cecal epithelial cell damage is a key factor in host injure during the development of E. tenella. The intracellular free Ca2+ of the host cell is closely related to the invasion, development and proliferation of intracellular parasites, and cell damage. To determine the relationship between Ca2+ and host cell damage in the schizogenic stage of E. tenella, we established a chick embryo cecal epithelial cells model of E. tenella infection. Fluorescence staining, flow cytometry, transmission electron microscopy, inhibition and blocking experiments were used to detect the damage effect and mechanism of host cells during the schizogenic stage of E. tenella. The results showed that the host cells cytoskeletal remodeling, cell and organelle structure was destroyed, and apoptosis and necrosis were increased during the schizont stage of E. tenella. Furthermore, the above-mentioned effects of the schizogenic stage of E. tenella on cells can be alleviated by reducing the intracellular Ca2+ concentration in the host cells. These observations indicate that the effect of host cell injury was closely related to Ca2+ during schizont stage of E. tenella.

Improved YOLOv4-tiny network for real-time electronic component detection.

In the electronics industry environment, rapid recognition of objects to be grasped from digital images is essential for visual guidance of intelligent robots. However, electronic components have a small size, are difficult to distinguish, and are in motion on a conveyor belt, making target detection more difficult. For this reason, the YOLOv4-tiny method is used to detect electronic components and is improved. Then, different network structures are built for the adaptive integration of middle- and high-level features to address the phenomenon in which the original algorithm integrates all feature information indiscriminately. The method is deployed on an electronic component dataset for validation. Experimental results show that the accuracy of the original algorithm is improved from 93.74 to 98.6%. Compared with other current mainstream algorithms, such as Faster RCNN, SSD, RefineDet, EfficientDet, and YOLOv4, the method can maintain high detection accuracy at the fastest speed. The method can provide a technical reference for the development of manufacturing robots in the electronics industry.

Serum Total Bilirubin Level Is Associated With Hospital Mortality Rate in Adult Critically Ill Patients: A Retrospective Study.

Background: Serum bilirubin level has been suggested to be associated with mortality for patients with severe sepsis. This study aimed to investigate the association of serum total bilirubin level with hospital mortality rate in adult critically ill patients. Method: Data were extracted from the Medical Information Mart for Intensive Care-III (MIMIC-III) database. Patients with measured serum total bilirubin levels that recorded within 24 h after admission were involved in this study. Association of serum total bilirubin level and hospital mortality rate was assessed using logistic regression analysis. Propensity score-matching (PSM) was used to minimize differences between different groups. Results: A total of 12,035 critically ill patients were herein involved. In patients with serum total bilirubin level ≥ 2 mg/dL, the hospital mortality rate was 31.9% compared with 17.0% for patients with serum total bilirubin level < 2 mg/dL (546/1714 vs. 1750/10321, P < 0.001). The results of multivariable logistic regression analysis showed that the odds ratio of mortality in patients with serum total bilirubin level ≥ 2 mg/dL was 1.654 [95% confidence interval (CI): 1.307, 2.093, P < 0.001]. After propensity score matching, in patients with serum total bilirubin level ≥ 2 mg/dL, the weighted hospital mortality rate was 32.2% compared with 24.8% for patients with serum total bilirubin level < 2 mg/dL, P = 0.001). Conclusions: Serum total bilirubin concentration was found to be independently associated with hospital mortality rate in adult critically ill patients.

IFN-γ induces senescence-like characteristics in mouse bone marrow mesenchymal stem cells.

BACKGROUND: Mesenchymal stem cells (MSC) are considered promising in tissue repair and regeneration medicine due to their proliferation and differentiation ability. Many properties of MSC are affected by cytokines, and IFN-γ has been shown to regulate MSC in many aspects. Senescence affects the proliferation, differentiation and cytokine secretion of MSC. OBJECTIVES: To investigate the effects of IFN-γ on the senescence-associated properties of MSC. MATERIAL AND METHODS: The MSC used in our study were isolated from the bone marrow (BM) of mice. Cell vitalities were measured by CCK8. The phenotypes and ROS of mBM-MSC were analyzed by flow cytometry. Cellular senescence was detected using SA-ß-gal stains. IL-6 and CXCL1 secretions were measured by ELISA. RESULTS: mBM-MSC can differentiated into osteocytes and adipocytes. They expressed CD29, CD106, and Sca-1, and did not express CD31, CD45 or FLK1. Our study showed that the cell vitalities of mBM-MSC were significantly reduced after IFN-γ treatment for 5 days, and the cell numbers were obviously lower after IFN-γ treatment for 5, 10 or 15 days. The IFN-γ group increased SA-ß-gal-positive cells and reactive oxygen species (ROS) significantly after 15 days of IFN-γ treatment. Moreover, IL-6 and CXCL1 secretions were upregulated by IFN-γ. CONCLUSIONS: Our study shows IFN-γ can induce senescence-like characteristics in mBM-MSC, suggesting a novel target for anti-aging therapy.

Development and validation of a solid-phase extraction method coupled with HPLC-UV detection for the determination of biogenic amines in Chinese rice wine.

A reliable and accurate method for the determination of seven biogenic amines (BAs) was developed and validated with Chinese rice wine samples. The BAs were derivatised with dansyl chloride, cleaned up using solid-phase extraction (SPE) and separated by high-performance liquid chromatography (HPLC) coupled with ultraviolet (UV) detection. The optimised derivatisation reaction, conducted at pH 9.6 and 60°C for 30 min, ensured baseline separation and peak symmetry for each BA. SPE clean-up using Oasis MCX cartridges yielded good recovery rates for all BAs and effectively reduced matrix effects. The developed method shows good linearity with determination coefficients of more than 0.9989 over a concentration range of 0.1-100 mg l-1. The limits of detection (LODs) for the investigated BAs ranged from 2.07 to 5.56 µg l-1. The intra- and inter-day relative standard deviations (RSDs) ranged from 0.86% to 3.81% and from 2.13% to 3.82%, respectively. Spiking experiments showed that the overall recovery rates ranged from 85% to 113%. Thus, the proposed method was demonstrated as being suitable for simultaneous detection, with accurate and precise quantification, of BAs in Chinese rice wine.

Association of HTR2A T102C and A-1438G polymorphisms with susceptibility to major depressive disorder: a meta-analysis.

Serotonin 2A receptor (HTR2A) gene was implicated to be associated with major depressive disorder (MDD) susceptibility due to its role of key neurotransmitter in many physiologic processes. A great number of related studies reported in different populations have emerged. The results of these studies, however, have been inconsistent and thereby definite conclusions are difficult to establish. With the cumulative data in recent years, it was necessary to carry out a comprehensive analysis of previous findings. Electronic databases were systematically searched for studies published before May 2013. Pooled odds ratios (OR) and 95 % confidence interval (CI) were estimated under three different genetic models. Subgroup and sensitivity analyses were also performed. A total of 21 studies, 3,299 patients and 4,092 controls, met the selection criteria. 15 studies included HTR2A T102C polymorphism (with a total of 2,409 patients and 3,130 controls), and 9 studies included HTR2A A-1438G polymorphism (with a total of 1,510 patients and 2,281 controls). Our results showed that no significant association of MDD susceptibility with T102C polymorphism was found in allelic analysis and genotypic analysis (For T vs. C: OR = 1.06, 95 % CI = 0.95-1.18, P = 0.307; For TT + TC vs. CC: OR = 1.07, 95 % CI = 0.90-1.28, P = 0.451; For TT vs. TC + CC: OR = 1.08, 95 % CI = 0.95-1.22, P = 0.235). With respect to A-1438G polymorphism, however, carriers with A allele tend to suffer from MDD (AA + AG vs. GG: OR = 1.20, 95 % CI = 1.02-1.43, P = 0.030). When stratified by race for T102C polymorphism and A-1438G polymorphism of the HTR2A, we found no significant association. In conclusions, our study suggests that the A allele of A-1438G polymorphism might play a role in susceptibility to MDD. On the contrary, T102C polymorphism does not seem to be capable of modifying MDD risk.

Left-behind children in rural China experience higher levels of anxiety and poorer living conditions.

AIM: The number of rural Chinese parents who leave their children with family members to work in the cities has increased dramatically over the last decade. This study compared the social anxiety of left-behind children (LBC) and children not left behind (non-LBC). METHODS: We investigated 1694 LBC and 1223 non-LBC, aged seven to 17 years, in a Chinese province using a structured questionnaire that included questions about socio-demographic characteristics, social anxiety, family function, quality of life, neglect and physical abuse. RESULTS: LBC displayed higher social anxiety scores, more neglect, lower parental educational level, lower quality of life, lower family function and lower prevalence of physical abuse than non-LBC. Multiple linear regression models showed that higher Social Anxiety Scales for Children (SASC) scores in LBC were clearly associated with lower quality of life, poorer family function, physical abuse, being female, having more siblings and minorities. In non-LBC, they were associated with lower quality of life, poorer family function, neglect, being female and physical abuse. CONCLUSION: LBC have a relatively higher level of social anxiety and poorer living conditions than non-LBC, and there are differences in social anxiety, and its relevant factors, between the two groups.

Association between MTHFR C677T polymorphism and depression: An updated meta-analysis of 26 studies.

BACKGROUND: Previous studies concerning the association between the 5,10-methylenetetrahydrofolate reductase (MTHFR) C677T gene polymorphism and depression have provided inconclusive findings. A meta-analysis was therefore performed to investigate a more reliable estimate. METHODS: This meta-analysis recruited 26 published studies which were selected by a search of electronic databases up to January 2013, including 4992 depression cases and 17,082 controls. Odds ratios (ORs) with 95% confidence intervals (CIs) were used to assess the association between MTHFR C677T polymorphism and depression susceptibility using random effect models. RESULTS: Meta-analyses results suggested that MTHFR C677T polymorphism contributed to the increased depression risk in overall populations (for T vs. C: OR=1.19, 95%CI=1.07-1.32; for TT+CT vs. CC: OR=1.15, 95%CI=1.01-1.31; for TT vs. CC: OR=1.42, 95%CI=1.16-1.75; for TT vs. CT+CC: OR=1.38, 95%CI=1.16-1.63). Subgroup analysis by ethnicity indicated an association in Asian populations (for T vs. C: OR=1.36, 95%CI=1.11-1.66; for TT+CT vs. CC: OR=1.32, 95%CI=1.03-1.69; for TT vs. CC: OR=1.88, 95%CI=1.26-2.79; for TT vs. CT+CC: OR=1.76, 95%CI=1.30-2.38); and a marginal association in White populations (for TT vs. CT+CC: OR=1.15, 95%CI=1.01-1.31). However, the association between the MTHFR C677T polymorphism and depression was not observed in the elderly. CONCLUSION: The MTHFR C677T polymorphism was associated with an increased risk of depression, especially in Asian populations. However, there was no evidence indicating a correlation in the elderly.

The TRIMCyp genotype in four species of macaques in China.

The tripartite motif protein (TRIM)5α/CypA fusion protein TRIMCyp in Old World monkeys is generally considered unable to restrict HIV-1 replication. Monkeys with TRIMCyp can serve as a unique animal model for studies of HIV-1 infection. The present study investigated the distribution and expression status of TRIMCyp in four species of macaques originating from China and its borderlands: pigtail macaques (Macaca nemestrina), rhesus macaques (Macaca mulatta), long-tailed macaques (Macaca fascicularis), and Tibetan macaques (Macaca thibetana). The results revealed that the frequencies of the TRIMCyp genotype were significantly different among different species and even within different populations of the same species. Interestingly, the TRIMCyp genotype was more prevalent among macaques originating from Yunnan and surrounding regions than those from other regions of China. Importantly, TRIMCyp individuals were first identified in Chinese M. mulatta originating from Yunnan, although multiple earlier studies failed to find CypA retrotransposition in this subspecies. Furthermore, TRIMe7-CypA, one of the splicing isoforms of the TRIMCyp transcript was expressed in M. nemestrina and M. mulatta but not M. fascicularis. The intra- and interspecies polymorphisms in the deduced TRIMCyp amino acid sequences of these macaques were also analyzed. Taken together, the data in this study provide important information about the genomic background of TRIMCyp among major species of Chinese macaques.

Short form 36-Item Health Survey test result on the empty nest elderly in China: a meta-analysis.

The aim of this study was to evaluate the quality of life of the empty nest elderly in China using meta-analytic techniques. Electronic databases were searched for studies concerning the quality of life of the empty nest elderly published before December 2011. Pooled weighted mean difference (WMD) and 95% confidence interval (95% CI) were estimated with fixed and random effect models. Subgroup analyses, sensitivity analyses and publication bias were also performed. Ten studies (6948 empty nesters and 4560 non-empty nesters) were included in this meta-analysis. Vitality (WMD -1.946; 95% CI -3.183 to -0.708), role emotional (WMD -6.084; 95% CI -8.545 to -3.623) and mental health (WMD -2.606; 95% CI -3.964 to -1.249) were statistically significantly low in the empty nest elderly whereas physical functioning, role physical, bodily pain, general health, and social functioning showed no statistical significance. Subgroup analyses showed a statistical significance with WMD of role emotional and mental health in urban area, that of role physical, bodily pain, vitality, role emotional and mental health in rural area, and that of vitality, social functioning, role emotional and mental health in mix area. This meta-analysis indicated that among eight dimensions of the Short Form 36-Item Health Survey (SF-36), only vitality, role emotional and mental health (mental components) were statistically significantly low in the empty nest elderly. More studies are needed to confirm this finding.

Interleukin 1 beta -511 C/T gene polymorphism and susceptibility to febrile seizures: a meta-analysis.

One previous meta-analysis found no evidence that interleukin 1 beta (IL-1ß) -511 gene polymorphism was associated with febrile seizures (FS) by pooling a limited number of studies. However, it is necessary for the meta-analysis to reevaluate the relationship with more recent findings. Electronic databases were systematically searched for studies published before June 2011. Pooled odds ratios (OR) and 95% confidence interval (CI) were estimated by means of a genetic model free approach. Subgroup and sensitivity analyses were also performed. All statistical analyses were conducted using Stata 9.0. A total of eight studies, 728 FS cases and 1,223 controls, met the selection criteria. The results show a significant association between IL-1ß -511 C/T gene polymorphism and FS (recessive genetic model TT vs. CC + CT: OR = 1.361, 95% CI: 1.065-1.738, P = 0.014). Subgroup analyses show a significant association in Asia (OR = 1.394, 95% CI: 1.005-1.935, P = 0.047), but not in Europe (OR = 1.387, 95% CI: 0.750-2.565, P = 0.298). IL-1ß -511 C/T gene polymorphism may play a role in susceptibility to FS, especially in Asia. Geographic differences may be a critical factor in the risk of FS.

An attenuated EIAV vaccine strain induces significantly different immune responses from its pathogenic parental strain although with similar in vivo replication pattern.

The EIAV (equine infectious anemia virus) multi-species attenuated vaccine EIAV(DLV121) successfully prevented the spread of equine infectious anemia (EIA) in China in the 1970s and provided an excellent model for the study of protective immunity to lentiviruses. In this study, we compared immune responses induced by EIAV(DLV121) to immunity elicited by the virulent EIAV(LN40) strain and correlated immune responses to protection from infection. Horses were randomly grouped and inoculated with either EIAV(DLV121) (Vaccinees, Vac) or a sublethal dose of EIAV(LN40) (asymptomatic carriers, Car). Car horses became EIAV(LN40) carriers without disease symptoms. Two of the four Vac horses were protected against infection and the other two had delayed onset or reduced severity of EIA with a lethal EIAV(LN40) challenge 5.5 months post initial inoculation. In contrast, all three Car animals developed acute EIA and two succumbed to death. Specific humoral and cellular immune responses in both Vac and Car groups were evaluated for potential correlations with protection. These analyses revealed that although plasma viral loads remained between 10(3) and 10(5)copies/ml for both groups before EIAV(LN40) challenge, Vac-treated animals developed significantly higher levels of conformational dependent, Env-specific antibody, neutralizing antibody as well as significantly elevated CD4(+) T cell proliferation and IFN-γ-secreting CD8(+) T cells than those observed in EIAV(LN40) asymptomatic carriers. Further analysis of protected and unprotected cases in vaccinated horses identified that cellular response parameters and the reciprocal anti-p26-specific antibody titers closely correlated with protection against infection with the pathogenic EIAV(LN40). These data provide a better understanding of protective immunity to lentiviruses.

A proviral derivative from a reference attenuated EIAV vaccine strain failed to elicit protective immunity.

To investigate essential factors that determine the efficacy of vaccines against lentiviruses, an effective attenuated equine infectious anemia virus (EIAV) vaccine strain and a proviral derivative of the vaccine were compared with respect to differences in inducing protective immunity. Although these two strains replicated equally well in vitro and in vivo, the proviral strain induced significantly less protection from disease and infection caused by viral challenge and significantly lower specific neutralizing capability. These findings indicated that the proviral strain had lost the ability to stimulate immune protection compared to the parental vaccine strain. A further analysis of the envelope gp90 gene variation revealed that compared to the proviral strain, the vaccine strain displayed a wide sequence diversity in immunogen composition. Thus, we inferred that the differences in immunogen composition might be the major cause for the failure of the proviral derivative to elicit the immune protection induced by the parental strain.

Class-specific molecularly imprinted polymers for the selective extraction and determination of sulfonylurea herbicides in maize samples by high-performance liquid chromatography-tandem mass spectrometry.

A novel method based on the molecularly imprinted solid-phase extraction (MISPE) procedure has been developed for the simultaneous determination of concentrations of sulfonylurea herbicides such as chlorsulfuron (CS), monosulfuron (MNS), and thifensulfuron methyl (TFM) in maize samples by liquid chromatography-tandem quadrupole mass spectrometry (LC-MS/MS). The molecularly imprinted polymer (MIP) for sulfonylurea herbicides was synthesized by precipitation polymerization using chlorsulfuron as the template molecule, 2-(diethylamino)ethyl methacrylate (DEAMA) as the functional monomer, and trimethylolpropane trimethacrylate (TRIM) as the cross-linker. The selectivities of the chlorsulfuron template and its analogs on the molecularly imprinted polymer were evaluated by high-performance liquid chromatography (HPLC). The extraction and purification procedures for the solid-phase extraction (SPE) cartridge with a molecularly imprinted polymer as the adsorbent for the selected sulfonylurea herbicides were then established. A molecularly imprinted solid-phase extraction method followed by high-performance liquid chromatography-tandem mass spectrometry for the determination of chlorsulfuron, monosulfuron, and thifensulfuron methyl was also established. The mean recoveries of these compounds in maize were in the range 75-110% and the limits of detection (LOD) of chlorsulfuron, monosulfuron, and thifensulfuron methyl were 0.02, 0.75, and 1.45 microg kg(-1), respectively. It was demonstrated that the MISPE-HPLC-MS/MS method could be applied to the determination of chlorsulfuron, monosulfuron, and thifensulfuron methyl in maize samples.

[Study of the correlation between the plasma viral load and protective immunity induced by the equine infectious anemia attenuated vaccine and its parental virulent strain].

The threshold hypothesis of attenuated lentiviral vaccine considers that the type of host response to infections of lentiviruses depends on the viral load. To evaluate the correlation between viral loads of the attenuated vaccine strain of equine infectious anemia virus (EIAV) and their effects to induce protective immunity, longitudinal plasma viral loads in groups of horses inoculated with either an attenuated EIAV vaccine strain (EIAV(DLV125)) or sub-lethal dose of an EIAV virulent strain (EIAV(LN40)) were compared. Similar levels of plasma viral loads ranging from 10(3)-10(5) copies/mL were detected from samples of these two groups of animals (P > 0.05) during 23 weeks post the inoculation. However, different responses to the challenge performed thereafter with lethal dose of the EIAV virulent strain were observed from the groups of horses inoculated with either EIAV(DLV125) or sub-lethal dose of EIAV(LN40). The protective efficiency was 67% (3 of 4 cases) and 0 (none of 2 cases), respectively. Our results implicate that the viral load of EIAV attenuated vaccine is not the primary factor, or at least not the solo primary factor, to determine the establishment of immune protection.

Application of the molecular combing technique to starch granules.

The molecular combing technique was used to dissociate the nanostructural units of starch granules from the starch fragments after a gelatinization process. With the help of atomic force microscopy (AFM), we observed that some nanostructural chains were just flowing out of the granules. It proves that there are substantive nanostructural units in the starch granules, a phenomenon not previously observed, so these nanostructural units were defined as suspected intermediates. Furthermore, we conclude that blocklets of starch granules are formed through twisting or distortion of nanochains.

[Comparison of proviral genomes between the Chinese EIAV donkey leukocyte-attenuated vaccine and its parental virulent strain].

The donkey leukocyte-attenuated vaccine of equine infectious anemia virus (EIAV) was the first lentiviral vaccine that induced solid protection from the infection of virulent strains. To elucidate the mechanism of increased immunogenicity and attenuated virulence of the vaccine, the proviral genomic DNA of an EIAV vaccine strain, EIAV(DLV121) was analyzed and compared with the genome of a parental virulent strain EIAV(DV117). Full length viral genomic DNAs were amplified as two segments by LA-PCR and were cloned. Because of the genomic diversity of retroviral quasispecies, 10 full-length sequences of EIAV(DLV121) and 4 full-length sequences of EIAV(DV117) from randomly picked clones were analyzed. Results showed that the average length of the complete nucleotide sequence of EIAV(DLV121) was 8,236bp and EIAV(DV117) was 8,249bp, with the inter-strain diversity of 2.8%. As for individual genes between the vaccine and virulent strains, the differences in nucleotide sequence of S2, LTR and env were significantly higher than the other genes with the diversity of 4.1%, 3.7% and 3.1%, respectively. Considerable variations in deduced amino acid sequences were found in S2, S3 and env. The diversities were 10.4%, 5.6% and 4.8%, respectively. Furthermore, the LTR of EIAV(DLV121) consisted of at least 5 subtypes grouped by their nucleotide sequences. There were two additional N-linked glycosylation sites in the deduced amino acid sequence of EIAV(DV117) gp90 than that of EIAV(DLV121). Among glycosylation sites in the gp90 of virulent strain, 3 were found unique only in EIAV(DV117), of which 2 were located in the principle neutralizing domain (PND). In addition, there was one EIAV(DLV121) -specific glycosylation site, which was positioned in the PND, too. Taken together, it is clear that greatly increased genomic diversity exists in the EIAV vaccine strain, which provides important information for the further study on biological characters of the Chinese EIAV attenuated vaccine.