Antigenicity, epitope mapping, and intracellular distribution of the NSP7α protein of porcine reproductive and respiratory syndrome virus.

Porcine reproductive and respiratory syndrome virus (PRRSV) is an important pathogen that causes huge economic losses to the global pig industry. Nonstructural protein 7α (NSP7α) of PRRSV is highly conserved among different lineages of PRRSV and could be a potential target for the development of detection methods. In this study, NSP7α was expressed in prokaryote (Escherichia coli) and purified. An NSP7α-ab-ELISA detection method was established, the NSP7α-ab-ELISA has 93.1 % coincidence rate with IDEXX PRRS X3 ab test kit. NSP7α antibody was detected in pig serum by ELISA 14 days following PRRSV infection. Three monoclonal antibodies (4H9, 3F2, and C10) against NSP7α prepared by a hybridoma technique were used for epitope mapping by indirect immunofluorescence. The 4H9, 3F2, and C10 antibodies all recognized the C-terminal 72-149 amino acid region of NSP7α. 4H9 reacted with amino acids 135-143, but 3F2 and C10 did not react with any truncated polypeptide. In addition, by using the monoclonal antibodies, NSP7α was localized solely in the cytoplasm, while the N protein was distributed in the cytoplasm and nucleus. The collective findings of the antigenicity and epitope of NSP7α will be helpful for understanding the antigenicity of NSP7α and developing PRRSV diagnostic methods.

T817MA Regulates Mitochondrial Dynamics via Sirt1 and Arc Following Subarachnoid Hemorrhage.

Spontaneous subarachnoid hemorrhage (SAH) is an acute neurologic emergency with poor outcomes, and mitochondrial dysfunction is known as one of the key pathological mechanisms underlying the SAH-induced early brain injury (EBI). 1-{3-[2-(1-benzothiophen-5-yl)ethoxy]propyl} azetidin-3-ol maleate (T817MA) is a newly synthesized neurotrophic compound that has been demonstrated to exert protective effects against brain injury. Here, we investigated the effect of T817MA in neuronal injury following experimental SAH both in vitro and in vivo. Primary cultured cortical neurons were treated with oxyhemoglobin (OxyHb) to mimic SAH in vitro, and T817MA at concentrations higher than 0.1 µM reduced OxyHb-induced neuronal injury. T817MA treatment significantly inhibited lipid peroxidation, reduced neuronal apoptosis and attenuated mitochondrial fragmentation. The results of western blot showed that T817MA markedly reduced the expression of mitochondrial fission proteins, fission protein 1 (Fis-1) and dynamin-related GTPase-1 (Drp-1), but prolonged the expression of the postsynaptic protein activity-regulated cytoskeleton-associated protein (Arc). In addition, T817MA significantly increased the expression of sirtuin 1 (Sirt1), which was accompanied by preserved enzymatic of isocitrate dehydrogenase (IDH2) and superoxide dismutase (SOD). Knockdown of Sirt1 and Arc via small interfere RNA (siRNA) transfection partially prevented the T817MA-induced protection in cortical neurons. Furthermore, treatment with T817MA in vivo significantly reduced brain damage and preserved neurological function in rats. The decreased expression of Fis-1 and Drp-1, as well as the increased expression of Arc and Sirt1 were also observed in vivo. Taken together, these data indicate that the neuroprotective agent T817MA protects against SAH-induced brain injury via Sirt1- and Arc-mediated regulation of mitochondrial dynamics.

Recombinant characteristics, pathogenicity, and viral shedding of a novel PRRSV variant derived from twice inter-lineage recombination.

Porcine reproductive and respiratory syndrome (PRRS), caused by the PRRS virus (PRRSV), is a significant threat to the global pig industry. In this study, a novel recombinant PRRSV, SD043, was isolated from a pig farm experiencing disease in 2019. Phylogenetic analysis revealed that SD043 belonged to lineage 1 of PRRSV-2 while recombination analyses revealed that it is a recombinant virus from lineage 1 and lineage 8 strains. Based on further analysis, SD043 underwent recombination twice. Pathogenicity studies revealed that SD043 causes mild clinical symptoms, thymus atrophy, and severe histopathological lesions in the lungs. Notably, virus shedding in SD043-infected piglets was detectable at 10 days post-inoculation with a high viral load in the respiratory or digestive tract, indicating that the recombinant PRRSV appears to shed higher numbers of virus. Furthermore, genomic surveillance based on all available PRRSVs circulating in Shandong province revealed an increasing increase in recombinant PRRSV since 2015, with the recombinant pattern (between lineages 1 and 8) being the same as that of SD043. These findings enable a better understanding of the process of twice recombination and virus shedding of recombinant PRRSV and can strengthen the prevention and control of the PRRSV epidemic.

Characterization of Two Immunodominant Antigenic Peptides in NSP2 of PRRSV-2 and Generation of a Marker PRRSV Strain Based on the Peptides.

Porcine reproductive and respiratory syndrome (PRRS) is a widespread disease with great economic importance in the pig industry. Although vaccines against the PRRS virus (PRRSV) have been employed for more than 20 years, differentiating infected from vaccinated animals remains challenging. In this study, all 907 non-structural protein 2 (NSP2) full-length sequences of PRRSV-2 available from GenBank were aligned. Two peptides, at positions 562-627 (m1B) and 749-813 (m2B) of NSP2, were selected, and their potential for use in differential diagnosis was assessed. Both m1B and m2B were recognized by PRRSV-positive pig serum in peptide-coated enzyme-linked immunosorbent assays. Further epitope identification yielded five overlapping short peptides for the immunodominant regions of m1B and m2B. Using the infectious clone of PRRSV HuN4-F112 as a template, the deletion mutants, rHuN4-F112-m1B, rHuN4-F112-m2B, and rHuN4-F112-C5-m1B-m2B, were generated and successfully rescued in Marc-145 cells. Growth kinetics revealed that the deletion of m1B and m2B did not significantly affect virus replication. Hence, m1B and m2B show potential as molecular markers for developing a PRRSV vaccine.

Rapid visual detection of porcine reproductive and respiratory syndrome virus via recombinase polymerase amplification combined with a lateral flow dipstick.

Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically devastating infectious diseases in the global swine industry. A rapid and sensitive on-site detection method for PRRS virus (PRRSV) is critically important for diagnosing PRRS. In this study, we established a method that combines reverse transcription recombinase polymerase amplification (RT-RPA) with a lateral flow dipstick (LFD) for detecting North American PRRSV (PRRSV-2). The primers and probe were designed based on the conserved region of all complete PRRSV-2 genomic sequences available in China (n = 512) from 1996 to 2020. The detection limit of the assay was 5.6 × 10-1 median tissue culture infection dose (TCID50) per reaction within 30 min at 42 °C, which was more sensitive than that of reverse transcription polymerase chain reaction (RT-PCR) (5.6 TCID50 per reaction). The assay was highly specific for the epidemic lineages of PRRSV-2 in China and did not cross-react with pseudorabies virus, porcine circovirus 2, classical swine fever virus, or porcine epidemic diarrhea virus. The assay performance was evaluated by testing 179 samples and comparing the results with those of quantitative RT-PCR (RT-qPCR). The results showed that the detection coincidence rate of RT-RPA and RT-qPCR was 100% when the cycle threshold values of RT-qPCR were < 32. The assay provides a new alternative for simple and reliable detection of PRRSV-2 and has great potential for application in the field.

The association between socioeconomic status and visual disability among older adults in China.

AIM: To investigate the association between socioeconomic status (SES) and visual disability (VD) among older Chinese adults. METHODS: We obtained data from the Second National Sample Survey on Disability, conducted in China in 2006. A total number of 192 375 older adults (aged≥65y) were screened for suspected VD via interviews with trained examiners. Those who screened positively for VD were referred to ophthalmologists to obtain a final diagnosis. RESULTS: VD was prevalent among 7.29% of Chinese adults aged 65 and older, and was higher in rural areas (8.71%) than in urban areas (4.82%). After adjusting for SES indicators and covariates, we found that less-educated older adults were more likely to suffer from VD, with an odds ratio (OR) of 2.50 (95%CI: 2.26-2.82) for illiterates, compared with those who graduated from senior high school or above. Older adults who were in the lowest income quintile were more at risk of VD, with an OR of 1.81 (95%CI: 1.68-2.95), compared with adults in the highest income quintile. In urban areas, when compared with adults who graduated from senior high school or above, those who did not continue their education after junior high school, primary school, or those who were illiterate, were more likely to suffer from VD, with an OR of 1.35 (95%CI: 1.51-1.59), 1.84 (95%CI: 1.60-2.12), and 2.63 (95%CI: 2.27-3.04), respectively. Lower levels of income were statistically significant when associated with VD. In rural areas, adults who were illiterate had an OR of 2.21 (95%CI: 1.75-2.79) when compared to adults with senior high school or above education level. Per capita, household income remained significantly associated with VD. Older adults who were ≥85, female, single, and residing in rural areas were associated with higher risks of VD. CONCLUSION: Individual-level SES among the elderly, in the form of education and income, is associated with VD among elderly Chinese adults in both urban and rural areas; however, the association is stronger in rural areas. Further studies are still required to explore the mechanism behind the relationships.

Evaluation of different surgical methods in treating recurrent patella dislocation after three-dimensional reconstruction.

PURPOSE: Recurrent patella dislocation (RPD) is the most common complication of patellar instability. The effects of different surgical techniques on the outcome of RPD treatments remain unclear. This study was conducted to compare the effects of three surgical techniques in treating RPD by three-dimensional (3D) reconstruction from computed tomography (CT) scans. METHODS: Sixty-eight patients with RPD and no previous surgical treatments who attended our hospital between October 2010 and 2013 were enrolled and randomly assigned into three groups: (1) medial patellofemoral ligament (MPFL) reconstruction and medial patellar retinaculum (MPR) plication group; (2) lateral patellar retinaculum (LPR) release and MPR plication group; and (3) MPFL reconstruction and LPR release group. Knee joints with flexion of 20° were scanned by a 64-row CT scanner and 3D reconstructed. Congruence angle (CA), patellar tilt angle (PTA), lateral patellofemoral angle (LPFA), and congruence of the lateral patellofemoral articular surface were measured. Knee joint function was evaluated by the Lysholm knee scoring scale, Kujala patellofemoral score, and International Knee Documentation Committee (IKDC) score. RESULTS: Pre-operative clinical characteristics were similar across groups. After treatment, the CA, PTA, and LPFA were reverted to normal post-operatively without statistically significant between-group differences. The MPFL reconstruction and LPR release group had the highest congruence of the lateral patellofemoral articular surface; while the (LPR) release and (MPR) plication group had the lowest Lysholm knee scoring scale, Kujala patellofemoral score, and IKDC score after surgery. CONCLUSIONS: The MPFL reconstruction and LPR release group had the best clinical outcomes among the three surgical methods, as indicated by better joint congruence after 3D joint reconstruction and higher knee function scores.

The complete mitochondrial genome of Juema pig Sus scrofa (Suina: Suidae) from southern Gansu.

Juema pig is a kind of rare and special pig which is well adapted to high altitude, cold climate and harsh natural environment. The complete mitochondrial genome of Juema pig Sus scrofa is a circular molecule of 16 532 bp in length, containing 13 protein-coding genes, two ribosomal RNAs, 22 transfer RNAs, and a control region. The A + T content of the overall base composition of H-strand is 60.7% (T: 26.2%; C: 26.0%; A: 34.5%; G: 13.3%). ND4L gene begins with GTG as start codon, ND2, ND3, and ND5 genes begin with ATA as a start codon, and other nine protein-coding genes start with ATG. Cyt b gene is terminated with AGA as stop codon, ND1 and ND2 genes are terminated with TAG as stop codon, COII, COIII, ND3, and ND4 end with T, while ATP6, ATP8, COI, ND4L, ND5, and ND6 end with TAA. In addition, the phylogenetic relationships from neighbor-joining analyses based on the 13 concatenated PCGs indicated (Tylopoda (Suina (Ruminantia (Hippopotamidae, Cetacea)))).

The complete mitochondrial genome of stretch spider Tetragnatha maxillosa (Araneae: Tetragnathidae).

The stretch spider Tetragnatha maxillosa (Araneae: Tetragnathidae) is found all over the world. In the present study, we investigated the complete mitochondrial genome of T. maxillosa and the mitogenome is a circular molecule of 14 414 bp in length, consists of 13 protein-coding genes, two ribosomal RNAs, 22 transfer RNAs, and a control region. The A + T content of the overall base composition of H-strand is 74.5% (T: 40.4%; C: 9.6%; A: 34.1%; G: 15.9%). COI gene begins with TTT as start codon, COII and COIII genes begin with TTG as start codon, ATP8, Cyt b, ND2, and ND4L genes begin with ATT as start codon, and other six protein-coding genes start with ATA. ATP6, ATP8, COI, COIII, ND1, ND3, and ND6 genes are terminated with TAA as stop codon, Cyt b, ND2, ND4, ND4L, and ND5 end with T, and COII ends with TAG. In addition, the phylogenetic relationships from neighbor-joining analyses based on the 13 concatenated PCGs indicated (Tetragnatha (Nephila (Argiope (Araneus, Neoscona)))).

Use of patient-specific templates in hip resurfacing arthroplasty: experience from sixteen cases.

PURPOSE: Hip resurfacing arthroplasty (HRA) is a technically demanding operation, requiring both accuracy and precision in placement of the acetabular and femoral components. Malalignment of the component can lead to notching and possible femoral neck fractures. We used specific templates created using a rapid prototyping machine based on the patients' anatomy, to aid in accurate intraoperative pin placement. METHODS: A 3D model of the hip was reconstructed using spiral computed tomography (CT) data by Amira 3.1 software in 16 patients in whom HRA was planned for hip osteoarthritis (OA). All of the patients in the study had normal contralateral hips. The rotational centre of femoral head on the normal side was superimposed using Imageware12.0 software to determine the centre of the femoral head on the contralateral side. The data was then used to produce patient-specific templates using a rapid prototyping technique. These templates were designed according to the anatomical features of femoral head surface, the rotation centre and the planned prosthesis shaft angle. The orientation of the prosthesis was determined by matching the model to the femoral head surface during the operation. In addition, a control group of 18 patients with OA was operated upon by the conventional method. RESULTS: The mean prosthesis stem shaft angle (SSA), as determined from postoperative imaging, was 138.68 ± 8.85° for the locating template group, and (118.9 ± 12.8) for the conventional group. CONCLUSIONS: The locating template designed and constructed preoperatively can provide precise and dependable location for hip resurfacing femoral components during arthroplasty and ensure the valgus stem placement necessary for optimal outcomes.

[Treatment of medial malleolus fractures with closed reduction and percutaneous internal fixation].

OBJECTIVE: To study clinical effects of minimally invasive, effective and economic operational method for the treatment of medial malleolus fractures. METHODS: From March 2008 to August 2010, 19 patients (12 males and 7 females, ranging in age from 17 to 42 years, averaged 31.7 years) with medial malleolus fractures were reviewed. Closed reduction and percutaneous internal fixation were applied, with a hollow compression screw inserted at the centre and perpendicularly to the fracture surface. A Kirschner wire was inserted through the cortical bone of opposite side and in accordance with the axis of inner malleolus. Postoperative therapeutic effect was evaluated by Kaikkonen sprained ankle scoring system and imageology examination. RESULTS: All the patients got primary healing of incisions and were followed up, the duration ranged from 6 to 30 months, with an average of 18.7 months. All the patients obtained bone union. Clinical healing time ranged from 2.6 to 3.8 months, averaged 3.2 months. According to Kaikkonen scoring system, the results were rated as excellent in 5 cases, good in 10 cases, moderate in 3 cases, and poor in 1 case. CONCLUSION: It is a minimally invasive, effective and economic method to treat medial malleolus fractures by closed reduction and percutaneous internal fixation with hollow compression screw and Kirschner wire.

[The relationship between TGFbeta1 and TIMP-1 and its effects on the diagnosis of hepatic fibrosis].

OBJECTIVE: To detect the expressions of serum transforming growth factor beta (TGFbeta1) and tissue inhibitor of metalloproteinase 1 (TIMP-1) in patients with chronic hepatitis B (CHB) in different phases of their liver fibrosis and to investigate the effects of TGFbeta1 and TIMP-1 on the diagnosis of hepatic fibrosis and the relationship between them. METHODS: Serum TGFbeta1 and TIMP-1 were detected by enzyme-linked immunosorbent assay and serum HA, LN, PIIINP were detected by radioimmunoassay in 40 CHB patients and 12 healthy persons. Liver biopsies were performed on the 40 patients. The relationships between the TGFbeta1 and TIMP-1 and hepatic fibrosis stages, hepatitis grades and the relevance of the five serum makers of the patients were analyzed. RESULTS: Serum TGFbeta1, TIMP-1 rose in the early stage of liver fibrosis and their sensitivities were higher than that of HA, LN and PIIINP. Serum levels of TGFbeta1 and TIMP-1 increased significantly with the development of fibrosis and had positive correlations with other fibrotic makers such as HA, LN and PIIINP. CONCLUSION: TGFbeta1 and TIMP-1 are involved in the course of developing hepatic fibrosis and TIMP-1 may be regulated by TGFbeta1. Both of them can be used as diagnostic markers of early hepatic fibrosis.