Search

This study investigated the effect of Luoshi Neiyi Formula(LSNYF) on hypoxia-inducible factor-1α(HIF-1α) and steroidogenic factor 1(SF-1) in endometriosis(EMs), aiming to explore the mechanism of Luoshi Neiyi Formula in treating EMs. Immunohistochemistry and quantitative real-time PCR(qPCR) were employed to determine the expression of HIF-1α and SF-1 in the endometriotic tissue. Human primary endometrial stromal cells(ESCs) were extracted and identified, in which the expression levels of HIF-1α and SF-1 were measured by immunofluorescence and qPCR. Cobalt chloride(CoCl_2) was used to simulate a low-oxygen environment in vitro, and the lentivirus with down-regulated expression of HIF-1α was constructed. The expression of HIF-1α or SF-1 was then determined by qPCR or Western blot. The effects of Luoshi Neiyi Formula-containing serum on the proliferation, migration, estradiol(E_2), HIF-1α, and SF-1 of ESCs in the low-oxygen environment were observed. Finally, the rat model of EMs with the syndrome of Qi stagnation and blood stasis was established by multi-factor intervention combined with autologous endometrium transplantation, and the effect of Luoshi Neiyi Formula on HIF-1α and SF-1 in the endometriotic tissue were studied. Compared with the normal endo-metrial tissue and ESCs, the endometriotic tissue and ectopic ESCs presented up-regulated mRNA and protein levels of HIF-1α and SF-1. Compared with the control group, treating eutopic ESCs with 50 μmol·L~(-1) CoCl_2 for 24 h promoted cell proliferation and up-regulated the mRNA level of HIF-1α and protein level of SF-1, while knocking down HIF-1α down-regulated the expression of SF-1 and E_2. Compared with the model group, the Luoshi Neiyi Formula-containing serum inhibited the proliferation and migration of ESCs and down-regulated the expression of E_2, HIF-1α, and SF-1. Similarly, the treatment with Luoshi Neiyi Formula attenuated the pathological injury of the endometriotic tissue and down-regulated the mRNA and protein levels of HIF-1α and SF-1 in the rat model of EMs. In summary, LSNYF may treat EMs by alleviating hypoxia and inhibiting the HIF-1α/SF-1 axis.

Human movement anomalies in indoor spaces commonly involve urgent situations, such as security threats, accidents, and fires. This paper proposes a two-phase framework for detecting indoor human trajectory anomalies based on density-based spatial clustering of applications with noise (DBSCAN). The first phase of the framework groups datasets into clusters. In the second phase, the abnormality of a new trajectory is checked. A new metric called the longest common sub-sequence using indoor walking distance and semantic label (LCSS_IS) is proposed to calculate the similarity between trajectories, extending from the longest common sub-sequence (LCSS). Moreover, a DBSCAN cluster validity index (DCVI) is proposed to improve the trajectory clustering performance. The DCVI is used to choose the epsilon parameter for DBSCAN. The proposed method is evaluated using two real trajectory datasets: MIT Badge and sCREEN. The experimental results show that the proposed method effectively detects human trajectory anomalies in indoor spaces. With the MIT Badge dataset, the proposed method achieves 89.03% in terms of F1-score for hypothesized anomalies and above 93% for all synthesized anomalies. In the sCREEN dataset, the proposed method also achieves impressive results in F1-score on synthesized anomalies: 89.92% for rare location visit anomalies ( τ = 0.5) and 93.63% for other anomalies.

TBR225 is one of the most popular commercial rice varieties in Northern Vietnam. However, this variety is highly susceptible to bacterial leaf blight (BLB), a disease caused by Xanthomonas oryzae pv. oryzae (Xoo) which can lead to important yield losses. OsSWEET14 belongs to the SWEET gene family that encodes sugar transporters. Together with other Clade III members, it behaves as a susceptibility (S) gene whose induction by Asian Xoo Transcription-Activator-Like Effectors (TALEs) is absolutely necessary for disease. In this study, we sought to introduce BLB resistance in the TBR225 elite variety. First, two Vietnamese Xoo strains were shown to up-regulate OsSWEET14 upon TBR225 infection. To investigate if this induction is connected with disease susceptibility, nine TBR225 mutant lines with mutations in the AvrXa7, PthXo3 or TalF TALEs DNA target sequences of the OsSWEET14 promoter were obtained using the CRISPR/Cas9 editing system. Genotyping analysis of T0 and T1 individuals showed that mutations were stably inherited. None of the examined agronomic traits of three transgene-free T2 edited lines were significantly different from those of wild-type TBR225. Importantly, one of these T2 lines, harboring the largest homozygous 6-bp deletion, displayed decreased OsSWEET14 expression as well as a significantly reduced susceptibility to a Vietnamese Xoo strains and complete resistance to another one. Our findings indicate that CRISPR/Cas9 editing conferred an improved BLB resistance to a Vietnamese commercial elite rice variety., Competing Interests: Authors XP, PD, HPT, HNT, and TB are employee of ThaiBinh Seed Corporation. This does not alter our adherence to PLOS ONE policies on sharing data and materials. There are no patents, products in development or marketed products associated with this research to declare.

A PCR-based survey of hemoprotozoa parasites detected Babesia bigemina, Theileria orientalis and Trypanosoma theileri among cattle and water buffalo in Vietnam, and a new Babesia sp. closely related to Babesia ovata was detected in cattle only. In addition, Theileria annulata and Trypanosoma evansi were not detected in both cattle and water buffalo. Phylogenetic analysis detected T. orientalis MPSP genotypes 3, 5, 7 and N3 in cattle and 5, 7, N1 and N2 in water buffalo. Additionally, water buffalo-derived T. theileri CATL sequences clustered together with a previously reported cattle-derived sequence from Vietnam. This is the first report of a new Babesia sp. in cattle, and T. orientalis MPSP genotype 7 and T. theileri in water buffalo in Vietnam.

Babesia bovis is the most virulent Babesia organism, resulting in a high mortality rate in cattle. The genetic diversity of B. bovis merozoite surface antigens (MSAs), such as MSA-1, MSA-2b, and MSA-2c, might be linked to altered immune profiles in the host animals. The present study aimed to develop type-specific PCR assays for Asian msa-1 genotypes, thereby re-analyzing the genetic diversity of msa-1 in Sri Lanka, Mongolia, and Vietnam. Specific primers were designed for nine Asian msa-1 genotypes, which had been detected based on the phylogeny constructed using msa-1 gene sequences retrieved from the GenBank database. Specificity of the type-specific PCR assays was confirmed using plasmids containing the inserts of msa-1 gene fragments that represent Asian genotypes. Furthermore, no amplicons were observed by these PCR assays when DNA samples of Babesia bigemina, Babesia ovata, Theileria annulata, Theileria orientalis, Trypanosoma evansi, Trypanosoma theileri, Anaplasma marginale, and Anaplasma bovis, and non-infected bovine blood were analyzed. In total, 109 B. bovis-positive blood DNA samples sourced from Sri Lanka (44 cattle), Mongolia (26 cattle), and Vietnam (23 cattle and 16 water buffaloes) were then screened by the type-specific PCR assays. The sequences derived from all of the PCR amplicons were phylogenetically analyzed. Out of 109 DNA samples, 23 (20 from cattle and 3 from water buffaloes) were positive for at least one genotype. In agreement with previous studies, five and four different genotypes were detected among the DNA samples from Sri Lanka and Vietnam, respectively. In contrast, four genotypes, including three novel genotypes, were detected from Mongolia. Five DNA samples were found to be co-infected with multiple genotypes. The sequences of the PCR amplicons clustered phylogenetically within the corresponding clades. These findings indicated that the type-specific PCR assays described herein are useful for the determination of genotypic diversity of the B. bovis msa-1 gene in Asia., (Copyright © 2015 Elsevier B.V. All rights reserved.)

In the present study, a total of 137 blood samples were collected from cattle and water buffaloes in central region of Vietnam and tested using nested polymerase chain reaction (nPCR), enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT) to determine the molecular and serological prevalence of Babesia bovis and Babesia bigemina. In cattle, the prevalence of B. bovis and B. bigemina was 21.3% and 16.0% by nPCR, 73.4% and 42.6% by ELISA and 60.6% and 59.6% by IFAT, respectively, whereas those of water buffalos were 23.3% and 0% by nPCR, 37.2% and 9.3% by ELISA and 27.9% and 18.6% by IFAT, respectively. IFAT and ELISA detected a higher number of infected cattle and water buffaloes than nPCR totally. Statistically significant differences in the prevalence of the two infections were observed on the basis of age. Overall, the current data suggest high incidence of B. bovis and B. bigemina infections in the central region of Vietnam, which is needed to develop comprehensive approach to the modern surveillance, diagnosis and control of bovine babesiosis.

We cloned two genes coding F107-C and K88-1NT fimbrial subunits from strains E. coli C and 1NT isolated from Thua Thien Hue province, Vietnam. The mature peptide of faeG gene from strain E. coli 1NT (called faeG-1NT) is 100 % similarity with faeG gene, while the CDS of fedA gene from strain C (called fedA-C) has a similarity of 97 % with the fedA gene. Expression of the faeG-1NT and fedA-C genes in E. coli BL21 Star™ (DE3) produced proteins of ~31 and 22 kDa, respectively. The effect of IPTG concentration on the K88-1NT and F107-C fimbriae production was investigated. The results showed that 0.5 mM IPTG is suitable for higher expression of K88-1NT subunit, while 0.75 mM IPTG strongly stimulated expression of F107-C subunit. The optimal induction time for expression was also examined. Generally, highest expression of K88-1NT subunit occurred after 6 h of induction, while that of F107-C subunit is after 14 h.

Hemoprotozoan infections often cause serious production losses in livestock. In the present study, we conducted a PCR-based survey of Babesia bovis, Babesia bigemina, Theileria annulata, Theileria orientalis, Trypanosoma evansi and Trypanosoma theileri, using 423 DNA samples extracted from blood samples of cattle (n=202), water buffaloes (n=43), sheep (n=51) and goats (n=127) bred in the Hue and Hanoi provinces of Vietnam. With the exception of T. annulata and T. evansi, all other parasite species (B. bovis, B. bigemina, T. orientalis and T. theileri) were detected in the cattle populations with B. bovis being the most common among them. Additionally, four water buffaloes and a single goat were infected with B. bovis and B. bigemina, respectively. The Hue province had more hemoprotozoan-positive animals than those from the Hanoi region. In the phylogenetic analyses, B. bovis-MSA-2b, B. bigemina-AMA-1 and T. theileri-CATL gene sequences were dispersed across four, one and three different clades in the respective phylograms. This is the first study in which the presence of Babesia, Theileria and Trypanosoma parasites was simultaneously investigated by PCR in Vietnam. The findings suggest that hemoprotozoan parasites, some of which are genetically diverse, continue to be a threat to the livestock industry in this country.

Babesia ovata is a tick-transmitted hemoprotozoan parasite that infects cattle. In our study, bovine blood samples (n=2,034) were collected from 10 different countries (Brazil, China, Ghana, Japan, Mongolia, the Philippines, South Africa, Sri Lanka, Thailand and Vietnam) and DNA extracted. The DNA samples were screened using an established and specific polymerase chain reaction (PCR) assay targeting the Apical membrane antigen 1 (AMA-1) gene. Parasite DNA was detected among samples collected from Japan, Mongolia and Thailand. Sequence analyses confirmed that the PCR assay detected only B. ovata AMA-1, and that amplicons from different geographical locations were conserved. Our findings highlight the importance of designing adequate strategies to control B. ovata infection in Japan, Mongolia, and Thailand.

Theileria orientalis is a benign bovine protozoan parasite that occasionally causes serious economic loss in the livestock industry. We report the findings of a molecular epidemiological survey of T. orientalis in 94 Vietnamese yellow cattle, 43 water buffaloes, 21 sheep, 21 goats and 85 blood-sucking ticks of cattle in the Thua Thien Hue province of Vietnam. The major piroplasm surface protein (MPSP) gene of T. orientalis was detected using polymerase chain reaction from 13 cattle (13.8%), 11 water buffaloes (25.6%), 1 sheep (4.8%) and 9 ticks (10.6%). Phylogenetic analysis using MPSP gene sequences showed the presence of seven genotypes, four previously categorized genotypes (Types 1, 3, 5 and 7) and three new genotypes (Types N-1, N-2 and N-3).

Rotavirus was examined from diarrheal stool samples of 158 infants in rural area near Ho Chi Minh City, Vietnam, from 1994 to 1996. Group A rotavirus was detected in 50%. G1 and G4 were the predominant serotypes. G3 was not detected. The most predominant type changed from year to year. Rotavirus was found in all seasons, especially in winter and autumn. Infants younger than 2 years of age were those mostly infected and the virus was suspected to invade high concentration in this area.

The production and role of endogenous cytokines during the course of secondary Corynebacterium (C.) pseudotuberculosis infection were investigated in mice. When immunized mice were challenged on day 28 after primary infection, tumor necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma) were found to appear at 3 hr and to reach the maximum at 24 hr after challenge. Spleen cells of mice primarily infected from 2 to 8 weeks before produced a significant amount of TNF-alpha and IFN-gamma when stimulated with formalin-killed bacteria. However, they could not produce detectable amounts of IL-4. The administration of anti-TNF-alpha monoclonal antibody (MAb) and IFN-gamma MAb increased bacterial proliferation in the organs of immune mice and exacerbated the secondary infection. Injection of anti-CD4 MAb alone or anti-CD4 plus anti-CD8 MAbs resulted in significantly increased mortality and a marked suppression of bacterial elimination as well as cytokine production of secondarily infected mice, while the treatment with anti-CD8 MAb alone showed no effect on either the resistance or cytokine production of mice. These results suggest that CD4, probably Th1 T cells, play an important role for establishment of protective immunity against secondary C. pseudotuberculosis infection by secreting TNF-alpha and IFN-gamma.

The present study was performed to investigate the role of CR3, the type 3 complement receptor, in host defense against primary and secondary Corynebacterium (C.) pseudotuberculosis infection in mice. Treatment of mice with 5C6, an anti-CR3 monoclonal antibody (mAb), resulted in unrestricted multiplication of bacteria in the organs and dramatically increased mortalities of the infected mice. Histological examinations showed the inflammation, degeneration and necrosis of organs and revealed that the infection-enhancing effect of 5C6 mAb was associated with the failure of mice to focus mononuclear phagocytes at sites of bacterial multiplication. These results suggest that CR3 plays an important role in host defense against primary as well as secondary C. pseudotuberculosis infection in mice.

The production and roles of endogenous tumor necrosis factor alpha (TNF-alpha) and gamma interferon (IFN-gamma) in the infection of Corynebacterium (C.) pseudotuberculosis were investigated in mice. The maximum levels of TNF-alpha and IFN-gamma were detected on day 4 after infection. The administration of anti-TNF-alpha monoclonal antibody (mAb) as well as anti-IFN-gamma mAb increased bacterial proliferation in the organs, leading to the death of infected mice, but anti-IFN-gamma mAb showed a less marked effect than anti-TNF-alpha mAb. The suppressive effect of anti-TNF-alpha and anti-IFN-gamma mAbs on anticorynebacterial resistance was augmented by the simultaneous administration of these antibodies. Anti-TNF-alpha mAb was found to be highly effective when administered on day 0 and day 4, suggesting that TNF-alpha produced during the early stage of infection is critical for the generation of resistance. Histologically, many microabscesses, severe follicular swelling and lymphocyte destruction were observed in mice treated with anti-TNF-alpha or anti-IFN-gamma mAb. Injection of anti-CD4 or anti-CD8 mAb also resulted in significantly increased mortality and a marked suppression of IFN-gamma production, but had no effect on TNF-alpha production. Carrageenan also showed a marked effect on the exacerbation of infection. Taken together, these results suggest that endogenously produced TNF-alpha and IFN-gamma are both essential to the host defense against C. pseudotuberculosis infection and that these cytokines may have an additive effect.