Prevalence and Genetic Analysis of Porcine Circovirus 3 in China From 2019 to 2020.

Porcine circovirus type 3 (PCV3), a virus belonging to the Circoviridae family, is considered to be associated with respiratory and neurological signs, cardiac and multisystemic inflammation, reproductive failure, and porcine dermatitis and nephropathy syndrome-like disease in pigs (Sus scrofa). In this study, epidemiological and serological investigations of PCV3 in clinically healthy pigs from different regions of China were performed. Overall, 42.87% (1,101/2,568) of pigs were positive for PCV3 Cap antibody via indirect enzyme-linked immunosorbent assay, with a higher prevalence of PCV3 in multiparous sows (62.22%, 881/1,416) and fattening pigs (28.96%, 159/549) than in suckling piglets (8.96%, 32/357) and nursery pigs (11.79%, 29/246). Of the 2,568 samples, 255 were further tested for PCV3 DNA using real-time polymerase chain reaction, and 63.14% of these were positive, with nearly half having <10 virus copies. The PCV3 DNA and antibody positivity rates were high in the pig serum samples; however, the virus titers and antibody levels were both low, indicating that the humoral immune response of PCV3-infected pigs was weak or lagging, and persistent or repeated infections could occur. Additionally, the complete genomes of 23 PCV3 strains were sequenced and analyzed, which showed nucleotide identities of 98.5~100.0%, 98.6~100.0%, and 99.2~100.0% in the complete genome, open reading frame (ORF)2, and ORF1 sequences, respectively, and amino acid identities of 96.7~100.0% and 99.3~100.0% in the capsid and replicase proteins, respectively. Phylogenetic analysis based on ORF2 nucleotide sequences indicated that the PCV3 strains obtained in the present study could be classified into three sub-clades, with most strains clustered into clade 3c, indicating that PCV3c is the dominant subtype in the regions of China investigated. In general, the present study revealed a high prevalence and high genetic divergence of PCV3 among Chinese pig herds, and indicated that the potential effect of PCV3 on the pig industry may be a concern.

Circular RNA-CDR1as is involved in lung injury induced by long-term formaldehyde inhalation.

OBJECTIVE: Formaldehyde (FA) is known to induce lung injury, but the underlying molecular mechanism remains largely unclear. CDR1as is an important member of the circular RNAs (circRNAs) family and functions as miRNA sponges with gene-regulatory potential. Our earlier circRNA microarray data showed CDR1as was highly expressed in lung tissue exposed to FA. However, the mechanism of circRNA-CDR1as mediates the FA-exposed lung injury is still unclear. This study aimed to explore the role of CDR1as in lung injury. MATERIALS AND METHODS: In this study, FA was inhaled at doses of 0.5, 2.46, and 5 mg/m3, respectively. After exposure 8 weeks, lung histopathological examination, lung injury score, and IL-1ß in bronchoalveolar lavage fluid (BALF) were determined. The expressions of CDR1as, rno-miR-7b and Atg7 were detected and the potential interaction of circRNA/miRNA/mRNA was predicted by bioinformatics analysis, including drawing circRNA/miRNA/mRNA interaction network, GO and KEGG analysis. RESULTS: Our results indicated FA inhalation upregulated the expression of CDR1as in lung tissues in a dose-dependent manner while the expression of rno-miR-7b decreased and Atg7 increased. Moreover, the alteration of CDR1as was positively correlated with lung injury. DISCUSSION AND CONCLUSIONS: CircRNA/miRNA/mRNA prediction further explained the possible effect mechanisms of CDR1as. These data implicated that CDR1as might be a critical regulator involved in lung injury induced by FA.

Requirement of Rab21 in LPS-induced TLR4 signaling and pro-inflammatory responses in macrophages and monocytes.

Lipopolysaccharide (LPS) induces macrophage/monocyte activation and pro-inflammatory cytokines production by activating Toll-like receptor 4 (TLR-4) signaling. Rab GTPase 21 (Rab21) is a member of the Rab GTPase subfamily. In the present study, we show that LPS induced TLR4 and Rab21 association and endosomal translocation in murine bone marrow-derived macrophages (BMDMs) and primary human peripheral blood mononuclear cells (PBMCs). In BMDMs, shRNA-mediated stable knockdown of Rab21 inhibited LPS-induced expression and production of pro-inflammatory cytokines (IL-1ß, IL-6 and TNF-α). Conversely, forced overexpression of Rab21 by an adenovirus construct potentiated LPS-induced IL-1ß, IL-6 and TNF-α production in BMDMs. Further studies show that LPS-induced TLR4 endosomal traffic and downstream c-Jun and NFκB (nuclear factor-kappa B) activation were significantly inhibited by Rab21 shRNA, but intensified with Rab21 overexpression in BMDMs. Finally, in the primary human PBMCs, siRNA-induced knockdown of Rab21 significantly inhibited LPS-induced IL-1ß, IL-6 and TNF-α production. Taken together, we suggest that Rab21 regulates LPS-induced pro-inflammatory responses by promoting TLR4 endosomal traffic and downstream signaling activation.

[Value of CHCHD2 as a potential marker of non-small cell lung cancer: analysis of 60 cases].

OBJECTIVE: To investigate the expression of CHCHD2, a potential tumor marker, in tumor and adjacent tissues from patients with non-small cell lung cancer (NSCLC). METHODS: Immunohistochemistry was used to detect the expression and location of CHCHD2 in the tumor tissues from 60 patients with NSCLC and 35 adjacent tissues to analyze the correlation of CHCHD2 expression with the clinicopathological variables and overall survival of the patients. The expression profile of CHCHD2 mRNA in NSCLC was analyzed using Oncomine database. RESULTS: The positivity rate of CHCHD2 was significantly higher in the tumor tissues than in adjacent tissues in patients with NSCLC (75.0% vs 17.1%). CHCHD2 positivity in the tumor tissues was associated with lymph node metastasis, pathological TNM stage, and tumor grades but not with age, gender, or histological type of the tumors. Analysis using Oncomine database showed that CHCHD2 mRNA was expressed at significantly higher levels in NSCLC than in normal control group (P<0.05). Kaplan-Meier survival analysis showed that NSCLC patients with a positive expression of CHCHD2 had a significantly shorter overall survival time than those negative for CHCHD2 (P<0.05). CONCLUSION: As a potential tumor marker, CHCHD2 over-expression plays a role in the occurrence and progression of NSCLC and promotes tumor invasion and metastasis, and can potentially serve as an indicator for early diagnosis and prognostic evaluation of NSCLC.

Pyridostigmine protects against cardiomyopathy associated with adipose tissue browning and improvement of vagal activity in high-fat diet rats.

Obesity, a major contributor to the development of cardiovascular diseases, is associated with an autonomic imbalance characterized by sympathetic hyperactivity and diminished vagal activity. Vagal activation plays important roles in weight loss and improvement of cardiac function. Pyridostigmine is a reversible acetylcholinesterase inhibitor, but whether it ameliorates cardiac lipid accumulation and cardiac remodeling in rats fed a high-fat diet has not been determined. This study investigated the effects of pyridostigmine on high-fat diet-induced cardiac dysfunction and explored the potential mechanisms. Rats were fed a normal or high-fat diet and treated with pyridostigmine. Vagal discharge was evaluated using the BL-420S system, and cardiac function by echocardiograms. Lipid deposition and cardiac remodeling were determined histologically. Lipid utility was assessed by qPCR. A high-fat diet led to a significant reduction in vagal discharge and lipid utility and a marked increase in lipid accumulation, cardiac remodeling, and cardiac dysfunction. Pyridostigmine improved vagal activity and lipid metabolism disorder and cardiac remodeling, accompanied by an improvement of cardiac function in high-fat diet-fed rats. An increase in the browning of white adipose tissue in pyridostigmine-treated rats was also observed and linked to the expression of UCP-1 and CIDEA. Additionally, pyridostigmine facilitated activation of brown adipose tissue via activation of the SIRT-1/AMPK/PGC-1α pathway. In conclusion, a high-fat diet resulted in cardiac lipid accumulation, cardiac remodeling, and a significant decrease in vagal discharge. Pyridostigmine ameliorated cardiomyopathy, an effect related to reduced cardiac lipid accumulation, and facilitated the browning of white adipose tissue while activating brown adipose tissue.

The PPARγ agonist, rosiglitazone, attenuates airway inflammation and remodeling via heme oxygenase-1 in murine model of asthma.

AIM: Rosiglitazone is one of the specific PPARγ agonists showing potential therapeutic effects in asthma. Though PPARγ activation was considered protective in inhibiting airway inflammation and remodeling in asthma, the specific mechanisms are still unclear. This study was aimed to investigate whether heme oxygenase-1 (HO-1) related pathways were involved in rosiglitazone-activated PPARγ signaling in asthma treatment. METHODS: Asthma was induced in mice by multiple exposures to ovalbumin (OVA) in 8 weeks. Prior to every OVA challenge, the mice received rosiglitazone (5 mg/kg, p.o.). After the mice were sacrificed, the bronchoalveolar lavage fluid (BALF), blood samples and lungs were collected for analyses. The activities of HO-1, MMP-2 and MMP-9 in airway tissue were assessed, and the expression of PPARγ, HO-1 and p21 proteins was also examined. RESULTS: Rosiglitazone administration significantly attenuated airway inflammation and remodeling in mice with OVA-induced asthma, which were evidenced by decreased counts of total cells, eosinophils and neutrophils, and decreased levels of IL-5 and IL-13 in BALF, and by decreased airway smooth muscle layer thickness and reduced airway collagen deposition. Furthermore, rosiglitazone administration significantly increased PPARγ, HO-1 and p21 expression and HO-1 activity, decreased MMP-2 and MMP-9 activities in airway tissue. All the therapeutic effects of rosiglitazone were significantly impaired by co-administration of the HO-1 inhibitor ZnPP. CONCLUSION: Rosiglitazone effectively attenuates airway inflammation and remodeling in OVA-induced asthma of mice by activating PPARγ/HO-1 signaling pathway.

[Effectiveness and safety of azithromycin in the treatment of bronchial asthma: a meta-analysis].

OBJECTIVE: To evaluate the effectiveness and safety of azithromycin in treatment of bronchial asthma. METHODS: Reports of randomized controlled trials (RCTs) describing azithromycin for treatment of asthma published before December 2013 were searched in CNKI, WANFANG, PubMed and Medline databases. The data of the included RCTs were extracted and the data quality was evaluated by two assessors independently. Meta-analyses were performed with Revman 5.1 software. RESULTS: Eight RCTs were identified. Meta-analysis of the data showed that compared with the control group, azithromycin treatment significantly improved the patients' PEF (WMD=0.15, 95%CI=0.06-0.24, P=0.001), scores of asthma control test (ACT) (WMD=1.59, 95%CI=0.95-2.23, P<0.00001), and FEV1% (WMD=1.44, 95%CI=0.40-2.49, P=0.007), but the improvement of FEV1% was observed only in Chinese patients (WMD=1.48, 95%CI=0.40-2.57, P=0.007). The scores of asthma control questionnaire (WMD=0.07, 95%CI=-0.11-0.25, P=0.45) or asthma quality of life questionnaire (WMD=-0.06, 95%CI=-0.42-0.31, P=0.77) were not affected by azithromycin. No severe adverse events were reported in these included studies. CONCLUSION: Azithromycin for asthma treatment can improve PEF, ACT and FEV1% (in Chinese patients only) but shows no significant effect on the quality of life of the patients. Azithromycin is well tolerated and may therefore be beneficial as adjuvant therapy for asthma.

LKB1/AMPK/mTOR signaling pathway in non-small-cell lung cancer.

Links between cancer and metabolism have been suggested for a long time but compelling evidence for this hypothesis came from the recent molecular characterization of the LKB1/AMPK signaling pathway as a tumor suppressor axis. Besides the discovery of somatic mutations in the LKB1 gene in certain type of cancers, a critical emerging point was that the LKB1/AMPK axis remains generally functional and could be stimulated by pharmacological molecules such as metformin in cancer cells. In addition, AMPK plays a central role in the control of cell growth, proliferation and autophagy through the regulation of mTOR activity, which is consistently deregulated in cancer cells. Targeting of AMPK/mTOR is thus an attractive strategy in the development of therapeutic agents against non-small-cell lung cancer (NSCLC). In this review, the LKB1/AMPK/mTOR signaling pathway is described, highlighting its protective role, and opportunities for therapeutic intervention, and clinical trials in NSCLC.

[A Meta-analysis of efficacy and safety of sublingual immunotherapy on allergic asthma].

OBJECTIVE: To evaluate the efficacy and safety of sublingual immunotherapy(SLIT) in patients with allergic asthma in order to provide reliable evidence for clinical application of SLIT. METHODS: To search published articles of randomized controlled trials (RCTs) in allergic asthma from CNKI, WANFANG, Pubmed and Medline databases. The methodological quality of trials was assessed by Jadad-scale. The heterogeneity was examined by using Stata 11.0 software. Fixed effect model or random effect model was used to pool the data. The articles which could not be pooled were carried out by descriptive analysis. The Egger's and Begg's test were used to evaluate the publication bias. RESULTS: There were total 6 RCTs included in this text. Compared with control group, SLIT could significantly reduce asthma symptom scores (SMD = -0.89, 95%CI -1.36--0.43, P = 0.000) and asthma medication scores (SMD = -4.53, 95%CI -6.97--2.08, P = 0.000), but not forced expiratory volume (FEV1) of lung function(SMD = 0.19, 95%CI -0.02-0.41, P = 0.078), neither serum sIgE levels (SMD = 0.05, 95%CI -0.58-0.69, P = 0.870). There were no obvious adverse events reported after treatment of SLIT. No publication bias were indicated by Egger's and Begg's tests. CONCLUSION: SLIT significantly reduces asthma symptom scores and medication scores, suggesting that SLIT is a safe and effective approach of immunotherapy. However, it still needs more highly qualified studies of RCTs to prove.

RT-PCR test for detecting porcine sapovirus in weanling piglets in Hunan Province, China.

The prevalence of porcine sapovirus infection in weanling pigs was investigated in Hunan Province, China, between August 2006 and October 2007. A total of 153 diarrheic fecal samples from ten intensive pig farms from ten representative administrative regions in Hunan province were examined for porcine sapoviruses using RT-PCR. Twenty-two of 153 (14.37 %) samples were found to contain porcine sapoviruses. Phylogenetic analysis showed that all the porcine sapovirus isolates in Hunan Province belonged to the porcine sapovirus genogroup III. The results of the present investigation have implications for the control of porcine sapovirus infection in pigs in Hunan Province, China.

Inhibition of cGMP phosphodiesterase 5 suppresses matrix metalloproteinase-2 production in pulmonary artery smooth muscles cells.

1. It has been shown that the beneficial effects of phosphodiesterase (PDE) 5 inhibition on pulmonary hypertension (PH) are associated with the induction of vascular relaxation and suppression of the proliferation of pulmonary artery smooth muscle cells (PASMC). In the present study, we investigated whether PDE5 inhibition affects the production and/or secretion of matrix metalloproteinases (MMPs) in PASMC, resulting in extracellular matrix remodelling in the pulmonary vasculature and, thus, the development of PH. 2. Primary cultured PASMC were stimulated with endothelin (ET)-1 and MMP-2 production and RhoA activation were then determinded using gelatin zymography and a GTP-bound RhoA assay, respectively. The effects of the selective PDE5 inhibitor sildenafil and subsequent protein kinase G-specific inhibitor Rp-8Br-cGMPs on MMP-2 production and RhoA activation were further exmamined. 3. Endothelin-1 (1-1000 nmol/L) concentration-dependently stimulated MMP-2 production and/or secretion in primary cultured PASMC, with 100 nmol/L ET-1 causing a 2.41-fold increase in MMP-2 production compared with control (P < 0.01). This increase in MMP-2 production was accompanied by RhoA activation, which was abolished by preincubation of cells with 10 micromol/L Y27632, an inhibitor of Rho-associated kinase (ROCK). Furthermore, 10 micromol/L Y27632 abolished the ET-1-induced production of MMP-2. 4. The selective PDE5 inhibitor sildenafil (0.1-1 micromol/L) concentration-dependently reduced the increased MMP-2 production induced by 100 nmol/L ET-1. Specifically, in the presence of 1 micromol/L sildenafil, the 100 nmol/L ET-1-induced increase in MMP-2 production was only increased 1.3-fold over that of the control (P < 0.01 vs 100 nmol/L ET-1-stimulated cells). 5. Suppression of RhoA activation was found to mediate the inhibitory effect of sildenafil on ET-1-induced increases in MMP-2 production. Furthermore, the protein kinase G-specific inhibitor Rp-8Br-cGMPs reversed the inhibitory effects of sildenafil on RhoA activation and MMP-2 production. 6. The results of the present study indicate that PDE5 inhibition suppresses RhoA/ROCK-mediated MMP-2 production by PASMC, which may contribute to the regulation of pulmonary vascular remodelling. Thus, PDE5 inhibition may benefit patients with PH through multiple mechanisms of action.

[Characteristic analysis of NA gene of human influenza viruses (H3N2) isolated from 1996 to 2005 in China].

The NA genes of 395 strains of human H3N2 influenza virus isolated from 1996 to 2005 in China were sequenced, analyzed with bioinformatics tools. The NA nucleotide sequence of phylogenetic tree showed a main evolution branch with multiple short side branches. The strains in the same year may be divided into several branches. There was an obvious lag between vaccine strains recommended by WHO and the Chinese circulating strains in phylogenetic tree of the NA nucleotide. The result also showed no amino acid deletion and insertion in the NA. In NA antigen sites, where including residues 197-199 aa, 431-434 aa and 339-347aa the mutation was higher, in contrast, the residues including 153 aa, 328-336 aa, 367-370aa and 400-403 aa, the mutation was lower. Besides the antigenic determinant sites, there also had the other amino acid mutated highly, such as 18, 23, 30, 93, 143, 208, 216, 221, 249, 265, 267, 307, 385 and 437 aa, among them 143 and 267 mutation were higher than that in antigenic determinant sites, their biological significance are not clear yet. The neuraminidase active-site residues in NA were highly conservative and the same were the disulphide bond and the glycosylation sites in NA. In conclusion, our analysis provides some information for influenza prevention and control and the NA inhibitor medicine application.