FOCUS-PDCA can effectively optimize the critical value of test items: FOCUS-PDCA moze efikasno da optimizuje kriticnu vrednost ispitivanja.

Background: To optimize the critical value of test items using FOCUS-PDCA (find, organize, clarify, understand, select, plan, do, check and act), and to set the personalized critical value of the test for different departments. Methods: We searched for literature reporting on the critical value and FOCUS-PDCA published over recent 5 years in order to understand the significance and status quo of critical value and FOCUS-PDCA. We also collected and analyzed the critical value data of hospital tests performed in Sichuan province hospitals in 2019, which were later compared to data from 2020 to determine the FOCUSPDCA cycle. Results: The proportion of critical values in the whole hospital decreased from 3.5% before optimization to 2.5% to 3% after optimization. The critical values of ICU, hematology, nephrology, urology, and neonatal departments after optimization significantly decreased compared with those before optimization, while the critical values of cardiac surgery, emergency ICU, cardiology, and neurosurgery ICU showed no significant difference before and after optimization. Contrary, the critical values of the infection department after optimization significantly increased before optimization. Conclusions: FOCUS-PDCA can effectively optimize the critical value of test items, which is beneficial for rational utilization of medical resources.

Characterization of grass carp (Ctenopharyngodon idella) beta-defensin 1: implications for its role in inflammation control.

Considering that fish grows in a complex aquatic environment, there is an increasing interest in fish ß-defensins, which is an important group of antimicrobial peptides (AMPs). In this study, grass carp (Ctenopharyngodon idella) ß-defensin 1 (gcdefb1) was isolated using homology cloning technology. Tissue distribution assay showed that gcdefb1 transcripts were expressed with the highest levels in brain and liver, followed by some mucous tissues. To examine gcDefb1 bioactivities, the recombinant gcDefb1 proteins fused with thioredoxin tag protein (Trx) (Trx-Defb1) were induced for production in Escherichia coli Rosetta-gami2(DE3)pLysS under optimal expression conditions. The antibacterial activity of Trx-Defb1 against Aeromonas hydrophila was assessed and its minimum inhibitory concentration (MIC) was 36 µM. Interestingly, Trx-Defb1 significantly inhibited LPS-induced Tnf-α (gcTnf-α) secretion and nitric oxide production in grass carp head kidney monocytes/macrophages (HKM), although Trx-Defb1 alone had no effect. Our studies provide the first evidence of fish ß-defensin 1 engaging in both antimicrobial and inflammation suppression process.

The influence of porcine epidemic diarrhea virus on pig small intestine mucosal epithelial cell function.

Porcine epidemic diarrhea (PED) is a highly contagious, acute enteric tract infectious disease of pigs (Sus domesticus) caused by porcine epidemic diarrhea virus (PEDV). PED is characterized by watery diarrhea, dehydration, weight loss, vomiting and death. PEDV damages pig intestinal epithelial tissue, causing intestinal hyperemia and atrophy of intestinal villi, with formation of intestinal epithelial cell cytoplasmic vacuoles. Since pig small intestinal epithelial cells (IECs) are target cells of PEDV infection, IEC cells were utilized as a model for studying changes in cellular activities post-PEDV infection. Monitoring of Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities demonstrated that PEDV infection decreased these activities. In addition, IECs proliferation was shown to decrease after PEDV infection using an MTT assay. Moreover, IECs apoptosis detected by flow cytometry with propidium iodide (PI) staining was clearly shown to increase relative to the control group. Meanwhile, animal experiments indicated that PEDV virulence for IEC cells was greater than viral virulence for Vero cells, although this may be due to viral attenuation after numerous passages in the latter cell line. Collectively, these studies revealed viral pathogenic mechanisms in PEDV-infected IECs and offer a theoretical basis for PEDV prevention and control.

Four polymorphisms in the IL-22 gene and the risk of cancer: A meta-analysis.

OBJECTIVE: IL-22 plays an important role in cancer carcinogenesis. However, the association between IL-22 polymorphisms and cancer risk was inconclusive. The objective of the current study was to assess these associations by meta-analysis. METHODS: PubMed, EMbase, CNKI, VIP, and Wanfang databases were searched up to 31 January 2018. The results were screened according to the inclusion and exclusion criteria. The associations between polymorphisms and cancer risk were estimated by meta-analysis. All analyses were performed using the Revman5.3 software. RESULTS: A total of four polymorphisms (rs2227485, rs1179251, rs1179246, and rs1182844) in seven studies were included. The results of meta-analysis indicated that the rs1179251 polymorphism [OR = 1.46, 95% CI (1.17, 1.82), P = 0.0008 for GG+GC vs. CC] was associated with increased risk of cancer, while the rs2227485, rs1179246, and rs1182844 polymorphisms were not associated with cancer risk. CONCLUSIONS: The current meta-analysis suggests that IL-22 gene rs1179251 polymorphism may be a risk factor for cancer.

Frequency of Polycythemia and Other Abnormalities in a Tibetan Herdsmen Population Residing in the Kham Area of Sichuan Province, China.

INTRODUCTION: The Kham Tibetans are one of several Tibetan ethnic subgroups living in the Kham area of China. Because studies on the high-altitude adaptation of the Kham people are scant, the main aim of this study is to investigate whether the response to hypoxia, especially polycythemia status, in the Kham Tibetans is different from other Tibetan ethnic subgroups. METHODS: The primary investigation was conducted on 346 native Kham Tibetan adults (268 men and 78 women) from 3 herdsmen villages located in Hongyuan County situated at an altitude of greater than 3600 m. The participants were aged 46.2±14.1 (21-82; mean±SD with range) years. Anthropometric measurements such as weight, height, waist circumference, body mass index, and blood pressure, as well as laboratory blood tests such as glycosylated hemoglobin, hemoglobin, total cholesterol, triglycerides, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and uric acid were analyzed. RESULTS: The concentrations of hemoglobin were 171.3±12.9 (66-229) mg·L-1 and 151.4±16.4 (86-190) mg·L-1 in men and women, respectively. The frequency of polycythemia was found to be 25.5 and 21.8% in men and women, respectively. Polycythemia was found to be significantly associated with glycosylated hemoglobin concentrations, hypertension, and hyperuricemia (P=0.002, 0.023, and 0.009, respectively). CONCLUSIONS: There is a higher frequency of polycythemia in the Kham Tibetans when compared with reported studies from other Tibetan ethnic subgroups living on the Qinghai-Tibet plateau.

Genetic susceptibility of lung cancer in Chinese population: An overview of systematic reviews and meta-analyses.

BACKGROUND: Genetic susceptibility of lung cancer has been widely studied for Chinese population, and meta-analysis of candidate gene association studies has also been performed for those genes. However, the overall evidence has not been well recognized. OBJECTIVE: To investigate genetic association for the risk of lung cancer in Chinese. METHOD: An overview of systematic reviews and meta-analyses of candidate gene association studies for lung cancer in Chinese was performed up to August 10th , 2016. The AMSTAR tool was used to assess the quality of the included systematic reviews and meta-analyses. Bibliometric analysis was performed to analyze the characteristics of reviews. RESULTS: A total of 21 variants in 17 genes from 20 meta-analyses were included in this study. All 20 meta-analyses were published from 2011 to 2016. The quality scores of AMSTAR ranged from 3 to 7. All included genes were in the pathogenesis of lung cancer, such as the CYPs genes, GSTs genes, and base excision repair genes. Three polymorphisms were found to be associated with decreased risk of lung cancer for Chinese, 15 polymorphisms were found to be associated with increased risk of lung cancer for Chinese, but three polymorphisms were found to be not associated with lung cancer risk for Chinese. CONCLUSION: The current study supports the genetic risk factors of lung cancer for Chinese are more likely to be variants from genes that contribute to the etiology of lung cancer.

Risk factors for postoperative infection in Chinese lung cancer patients: A meta-analysis.

OBJECTIVE: The objective of this study was to identify risk factors for postoperative infection in Chinese lung cancer patients. METHODS: A comprehensive search was performed in PubMed, EMbase, CNKI, and WanFang Data databases to identify studies investigating risk factors for postoperative infection in Chinese lung cancer patients. Meta-analysis was performed by using Revman 5.2 software. RESULTS: A total of 12 studies were included. The results of meta-analysis showed that old age, male gender, diabetes, cigarette smoking, squamous cell carcinoma, pulmonary diseases, longer of mechanical ventilation, and longer time of surgery procedure were associated with increased risk of postoperative infection. In addition, prophylactic antibiotic was associated with decreased risk of postoperative infection. CONCLUSION: The current meta-analysis suggests that the old age, male gender, diabetes mellitus, cigarette smoking, squamous cell carcinoma, pulmonary diseases, longer of mechanical ventilation, and longer of surgery time are risk factors for postoperative infection in Chinese lung cancer patients. Due to the limited quality and quantity of included studies, more high-quality studies are needed to verify the above results.

The association between polymorphisms in the PDCD1 gene and the risk of cancer: A PRISMA-compliant meta-analysis.

BACKGROUND: The effects of the programed cell death 1 (PDCD1) gene polymorphisms on cancer risk have been investigated in some studies; however, the results were conflicting and ambiguous. Therefore, we aimed to do a meta-analysis to investigate the association of PDCD1 polymorphisms with cancer risk from all eligible case-control studies. MATERIALS AND METHODS: An electronic search of the PubMed, Embase, Chinese National Knowledge Infrastructure, and Wanfang databases was performed. The association between PDCD1 polymorphisms with cancer risk was calculated with odds ratios (ORs) and their corresponding 95% of confidence intervals (CIs). RESULTS: A total of 24 case-control studies from 13 articles that investigated the associations of 5 widely studied polymorphisms in PDCD1 gene and cancer risks were included. The results of meta-analysis: the PDCD-1.5 (rs2227981) and PDCD-1.3 (rs11568821) polymorphisms were associated with decreased risk of cancer (rs2227981: OR = 0.75, 95% CI: 0.64-0.86, P < 0.0001 for TT vs TC + CC; rs11568821: OR = 0.79, 95% CI: 0.65-0.96, P = 0.02 for TC vs TT), while no significant associations were found for the other 3 polymorphisms (PDCD-1.9 [rs2227982] polymorphism: OR = 1.03, 95% CI: 0.90-1.18, P = 0.66 for CC + TC vs TT; PDCD1 rs7421861 polymorphism: OR = 1.10, 95% CI: 0.96-1.25, P = 0.16 for CC + TC vs TT; PDCD-1.6 [rs10204525] polymorphism: OR = 0.93, 95% CI: 0.82-1.05, P = 0.24 for GG + GA vs AA). CONCLUSION: The meta-analysis suggests that the PDCD-1.5 (rs2227981) and PDCD-1.3 (rs11568821) polymorphisms are associated with susceptibility of cancer. Further studies with larger sample sizes are required to make a better assessment of the above association.

Functional expression and characterization of grass carp IL-10: an essential mediator of TGF-ß1 immune regulation in peripheral blood lymphocytes.

In mammals, interleukin-10 (IL-10) is known as a potent regulatory cytokine with pleiotropic effects on various leukocytes. Although teleost IL-10 has been identified in several fish species, its immunoregulatory role is still poorly understood. In the present study, grass carp IL-10 (gcIL-10) has been identified and recombinant gcIL-10 (rgcIL-10) was prepared by using a prokaryotic expression system. Using grass carp peripheral blood lymphocytes (PBLs) as a cell model, rgcIL-10 was shown to up-regulate the cell activity, which was consistent with the regulatory tone of transforming growth factor ß1 (TGF-ß1). An interesting question that follows would be whether IL-10 and TGF-ß1 redundantly regulate PBL activity. To address this question, we investigated the effect of IL-10 on the cell activity of grass carp PBLs challenged by TGF-ß1 using immunoneutralization, showing that removal of endogenous IL-10 could abolish TGF-ß1-induced cell activity, indicating an essential role of IL-10 in TGF-ß1 immune regulation. Furthermore, to examine the connection between TGF-ß1 and IL-10 during immunoregulatory process, effect of TGF-ß1 on IL-10 expression was investigated. Results showed that TGF-ß1 displayed stimulatory effects on gcIL-10 mRNA expression and protein secretion in the same cell model, suggesting that TGF-ß1 may function through its effect on IL-10 production in fish immunity. These results provide clues to the potential immunoregulatory role and production of IL-10 in teleost.

Evidence for pituitary adenylate cyclase-activating peptide as a direct immunoregulator in teleost head kidney.

In mammals, pituitary adenylate cyclase activating polypeptide (PACAP) is a potent anti-inflammatory factor, showing that it inhibits the expression and release of proinflammatory cytokines and enhances the production of anti-inflammatory factors. However, whether fish PACAP plays similar regulatory roles as seen in mammals remains unclear. In the present study, expression of PACAP-specific receptor PAC1-R was shown in grass carp head kidney and spleen, supporting that PACAP may have a direct effect on fish immune cells. To test this hypothesis, the immunoregulatory role of grass carp PACAP (gcPACAP) was examined in head kidney leucocytes (HKLs). Results showed that gcPACAP inhibited basal and further attenuated lipopolysaccharide (LPS)-stimulated cell viability of HKLs, indicating that gcPACAP may possess similar inhibitory property at cellular level as seen in mammals. Curiously, in vitro and in vivo studies revealed that gcPACAP stimulated proinflammatory factors (IL-1ß and TNF-α) but not IL-10 mRNA expression in HKLs and head kidney. Moreover, bacterial infection and LPS enhanced IL-1ß, TNF-α and IL-10 mRNA expression in grass carp head kidney and HKLs, respectively, and these stimulatory effects were not influenced by gcPACAP. These findings suggest that PACAP plays distinct roles, at least does not function as an anti-inflammatory factor, in fish compared with that in mammals.

Functional characterization of TNF-α in grass carp head kidney leukocytes: induction and involvement in the regulation of NF-κB signaling.

Tumor necrosis factor-alpha (TNF-α) is a potent regulatory cytokine, which serves as a key mediator of inflammation, immunity and apoptosis in mammals. Identification, expression and regulatory effects of TNF-α have been reported in various fish species, showing the structural and functional similarity or discrepancy between each other. In this study, TNF-α was identified from grass carp (Ctenopharyngodon idella) and the deduced grass carp TNF-α (gcTNF-α) protein possessed the TNF family signature motifs, a protease cleavage site, a transmembrane domain and two conserved cysteine residues. Further studies showed that gcTNF-α expression was induced with a rapid kinetics by immune challenge in vitro and in vivo. To characterize the function of gcTNF-α, recombinant gcTNF-α (rgcTNF-α) was prepared by using the Escherichia coli expression system. It was shown to enhance the mRNA expression of gcTNF-α and gcIL-1ß in head kidney leukocytes (HKLs), confirming the biological activity of rgcTNF-α. In the same model, NF-κB inhibitor (PDTC) was able to attenuate rgcTNF-α-induced gcTNF-α mRNA expression, implying the involvement of NF-κB pathway in fish TNF-α action. This notion was reinforced by the finding that rgcTNF-α could induce the phosphorylation of IκBα in a time-dependent oscillation in HKLs, indicating a dynamical variation of NF-κB activity as seen in mammals. In addition, rgcTNF-α could up-regulate the expression of two TNF receptor-associated factors (TRAF), TRAF1 and TRAF2, in a time- and dose-dependent manner, suggesting that gcTNF-α may function as a regulator of fish NF-κB pathway. These results for the first time reveal the link of gcTNF-α to the NF-κB pathway and provide a better understanding of TNF-α signaling in teleost immunity.

Characterization of Foxp3 gene from grass carp (Ctenopharyngodon idellus): a rapamycin-inducible transcription factor in teleost immune system.

In this study, we cloned grass carp foxp3 (gcfoxp3) gene including 5' flanking region and determined its expression profiles in vivo under immunosuppressive conditions. Sequence analysis revealed that the promoter of gcfoxp3 contains AP-1, AML-1/Runx1, NF-κb and GATA-3 binding sites, which positively or negatively regulate mammalian foxp3 expression. In addition, the intron II of gcfoxp3 contains some putative binding sites including AP-1, NFAT, Smad3, RAR, CREB/ATF and FOXO1, which are corresponding to their locations in the proximal intronic enhancers of human foxp3. In an in vivo model of grass carp, an immunosuppressive agent rapamycin was showed to stimulate gcfoxp3 mRNA expression in thymus, gill, head kidney and spleen after bacterial challenge. Moreover, rapamycin increased gcFoxp3 protein levels with an additive manner in the infected fish. These findings support the involvement of fish Foxp3 in immune response and highlight a possible signaling pathway that regulates teleost Foxp3 expression.

Regulation of dopamine D2 receptor expression in grass carp pituitary cells: a possible mechanism for dopaminergic modification of luteinizing hormone synthesis.

In many fishes, dopamine (DA) strongly inhibits luteinizing hormone (LH) release by direct action at the pituitary level. In this study, the effect of DA on LH synthesis was examined by detecting its ß-subunit mRNA level in immature grass carp pituitary cells. Results showed that DA inhibited LHß mRNA expression and its inhibition was antagonized by a DA D2 receptor (DRD2) antagonist, sulpiride, suggesting that DA inhibited LH synthesis via DRD2. This notion was further supported by the finding that the grass carp DRD2 (gcDRD2) immunoreactivity was observed in the proximal pars distalis of the pituitary in which gonadotrophs are distributed. Accordingly, a full-length cDNA for DRD2 was cloned from grass carp pituitary and it showed closer phylogenetic relationships to the DA D2 receptors compared with the D3 and D4 or D1-like receptors in other vertebrates. Besides brain, the expression of this receptor in pituitary was revealed by tissue distribution assay, implying the pituitary function of gcDRD2 in immature grass carp. In grass carp pituitary cells, gcDRD2 transcript level was stimulated by DA and this stimulation was blocked by sulpiride. However, hCG, a functional homolog of grass carp LH, was found to inhibit gcDRD2 mRNA expression, indicating an intrapituitary negative feedback of LH on gcDRD2 expression. In view of our observation that the DRD2 mediated the dopaminergic inhibition of LH synthesis, we speculate that the DA stimulation and LH suppression on gcDRD2 may reinforce or attenuate the DA inhibition on LH synthesis, respectively and this regulation of gcDRD2 may at least partially contribute to the steady state levels of LH mRNA in prepubertal grass carp.