Comparison of two novel diagnostic criteria for bronchopulmonary dysplasia in predicting adverse outcomes of preterm infants: a retrospective cohort study.

BACKGROUND: This study aimed to compare the predictive value of two diagnostic criteria for bronchopulmonary dysplasia (BPD) in preterm infants with gestational age (GA) < 32 weeks for death or severe respiratory morbidity at corrected age of 18-24 months. METHODS: In this retrospective cohort study, clinical data from July 2019 to September 2021 were classified by 2018 National Institute of Child Health and Human Development (NICHD) and 2019 Jensen definitions of BPD. Based on the follow-up results, the enrolled population was divided into adverse outcome group and normal outcome group. Logistic regression and receiver operating characteristic (ROC) curve analyses were conducted to explore the risk factors of adverse outcomes and evaluate the predictive value of both diagnostic criteria. RESULTS: Of 451 infants, 141 (31.3%) had adverse outcomes, which increased with increasing severity of BPD. Logistic regression analysis showed only BPD was an independent risk factor for adverse outcomes in preterm infants. ROC analysis revealed that both diagnostic criteria showed similar predictive values (2018 NICHD definition AUC = 0.771 vs. 2019 Jensen definition AUC = 0.770), with specificities of 93.5% and 96.8%, respectively; however, combining them separately with GA or birth weight did not improve their predictive values. CONCLUSIONS: The two novel definitions of BPD demonstrate similar predictive values in predicting death or severe respiratory morbidity at corrected age of 18-24 months, with higher specificity observed in both.

Effect of IL-8 on hepatocellular carcinoma-associated metastasis by targeting MMP9 in mice.

Background: Interleukin-8 (IL-8) and matrix metallopeptidase 9 (MMP9) are overexpressed in hepatocellular carcinoma (HCC), and both are related to tumor metastasis, but whether they regulate HCC metastasis is still unclear. Methods: HCC orthotopic implantation and colonization mice models were established in vivo. Model mice were treated with IL-8 and or MMP9 inhibitors, protein kinase C (PKC) inhibitors, or extracellular regulated protein kinases 1/2 (ERK1/2) inhibitors. Liver metastasis and lung metastasis of model mice were confirmed by hematoxylin and eosin staining. The population of circulating tumor cells (CTCs) was detected by flow cytometry. The expression of MMP9 in tumor tissues was determined by quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry. In vitro, HCC LM6 (HCCLM6) cells were treated with IL-8 combined PKC inhibitor or ERK1/2 inhibitor. The expression of MMP9 was confirmed by qRT-PCR and Western blot, and the activation of the PKC/ERK1/2 signaling pathway was confirmed by Western blot. Results: IL-8 promoted liver metastasis and lung metastasis in orthotopic transplantation model mice, increased the proportion of CTCs and promoted the expression of MMP9 in tumor tissues, but these effects were reversed by PKC inhibitor or ERK1/2 inhibitor. In vivo colonization experiments, IL-8 promoted tumor cell metastasis to the liver and lung, but the MMP9 inhibitor reversed the metastasis-promoting effect of IL-8. In cell experiments, IL-8 promoted the expression of p-PKC and p-ERK1/2 and inhibited the expression of PKC and ERK1/2; the promotion of MMP9 expression by IL-8 was reversed by PKC inhibitor or ERK1/2 inhibitor. Conclusions: IL-8 up-regulated the expression of MMP9 by activating the PKC/ERK1/2 signaling pathway, thereby promoting the metastasis and colonization of HCC.

Ultrasound-guided needle release of A1 pulley combined with corticosteroid injection is more effective than ultrasound-guided needle release alone in the treatment of trigger finger.

BACKGROUND: The purpose of the present study was to evaluate the clinical effectiveness of ultrasonography-guided needle release of A1 pulley combined with corticosteroid injection by comparing it with ultrasound-guided needle release of the A1 pulley alone. METHODS: A total of 49 patients (55 fingers, thumb) with trigger fingers were included in this retrospective study. Twenty-seven fingers were treated with ultrasound-guided needle release of the A1 pulley alone (monotherapy group), and 28 fingers were treated with needle release of the A1 pulley combined with corticosteroid injection (combination group). Visual analog scale (VAS), Froimson scale, postoperative recurrence rate, and thickness of A1 pulley at baseline, Week-2, Week-12, and Month-6 were recorded. RESULTS: Higher clinical cure rates were observed in the combination group at Week-2 after treatment among patients with the Froimson scale Grade III and IV (p < 0.05). Among Froimson scale Grade IV patients, the combination group had a significantly thinner thickness of A1 pulley and better articular pain relief at Week-2 (all p < 0.05). No significant differences were found in the clinical cure rate, the thickness of the A1 pulley, articular pain relief, and recurrence rate between the two groups at Week-12 and Month-6 (all p > 0.05). CONCLUSIONS: Ultrasonography-guided needle release of A1 pulley plus corticosteroid injection was superior to ultrasonography-guided A1 pulley needle release alone during early-stage treatment of severe patients with trigger fingers. Moreover, ultrasonography-guided A1 pulley needle release combined with corticosteroid injection narrows the thickness of the A1 pulley. It is necessary to carry out preoperative evaluation and individualized treatment for patients of various severities.

Modifiable Factors and Incident Gout Across Ethnicity Within a Large Multiethnic Cohort of Older Adults.

OBJECTIVE: Gout disproportionately affects older Pacific Islander and Black populations relative to White populations. However, the ethnic-specific determinants remain understudied within these groups, as well as within other ethnicities. We examined gout incidence and associations with behavioral factors, including diet, alcohol, and smoking, within a large multiethnic population of older adults from the Multiethnic Cohort Study, which linked prospective cohort data to Medicare gout claims between 1999-2016. METHODS: Using samples of Black (n = 12,370), Native Hawaiian (n = 6459), Japanese (n = 29,830), Latino (n = 17,538), and White (n = 26,067) participants, we conducted multiple Cox regressions, producing hazard ratios (HRs) and 95% CIs. RESULTS: Relative to White individuals, Native Hawaiians had the highest risk of gout (HR 2.21, 95% CI 2.06-2.38), followed successively by Black and Japanese participants, whereas Latino individuals had a lower risk of gout (HR 0.78, 95% CI 0.73-0.83). Alcohol use was associated with an increased risk, with significantly greater effects observed among Japanese participants drinking ≥ 3 drinks per day (HR 1.46, 95% CI 1.27-1.66), or > 5 beers per week (HR 1.29, 95% CI 1.17-1.43), compared to White individuals (Pinteraction < 0.001). Former smokers with ≥ 20 pack-years had an increased risk (HR 1.14, 95% CI 1.06-1.22). Higher dietary quality was associated with a decreased gout risk, with the largest effect observed among White participants (HRQ5vsQ1 0.84, 95% CI 0.79-0.90), whereas vitamin C was weakly associated with a decreased risk of gout only among Japanese individuals (HR 0.91, 95% CI 0.85-0.98). CONCLUSION: Overall, notable ethnic differences were observed in both gout risk and associations with modifiable behavioral factors. Our findings offer crucial insights that may improve precision in preventing and managing gout.

Targeting GATA1 and p2x7r Locus Binding in Spinal Astrocytes Suppresses Chronic Visceral Pain by Promoting DNA Demethylation.

Irritable bowel syndrome is a gastrointestinal disorder of unknown etiology characterized by widespread, chronic abdominal pain associated with altered bowel movements. Increasing amounts of evidence indicate that injury and inflammation during the neonatal period have long-term effects on tissue structure and function in the adult that may predispose to gastrointestinal diseases. In this study we aimed to investigate how the epigenetic regulation of DNA demethylation of the p2x7r locus guided by the transcription factor GATA binding protein 1 (GATA1) in spinal astrocytes affects chronic visceral pain in adult rats with neonatal colonic inflammation (NCI). The spinal GATA1 targeting to DNA demethylation of p2x7r locus in these rats was assessed by assessing GATA1 function with luciferase assay, chromatin immunoprecipitation, patch clamp, and interference in vitro and in vivo. In addition, a decoy oligodeoxynucleotide was designed and applied to determine the influence of GATA1 on the DNA methylation of a p2x7r CpG island. We showed that NCI caused the induction of GATA1, Ten-eleven translocation 3 (TET3), and purinergic receptors (P2X7Rs) in astrocytes of the spinal dorsal horn, and demonstrated that inhibiting these molecules markedly increased the pain threshold, inhibited the activation of astrocytes, and decreased the spinal sEPSC frequency. NCI also markedly demethylated the p2x7r locus in a manner dependent on the enhancement of both a GATA1-TET3 physical interaction and GATA1 binding at the p2x7r promoter. Importantly, we showed that demethylation of the p2x7r locus (and the attendant increase in P2X7R expression) was reversed upon knockdown of GATA1 or TET3 expression, and demonstrated that a decoy oligodeoxynucleotide that selectively blocked the GATA1 binding site increased the methylation of a CpG island in the p2x7r promoter. These results demonstrate that chronic visceral pain is mediated synergistically by GATA1 and TET3 via a DNA-demethylation mechanism that controls p2x7r transcription in spinal dorsal horn astrocytes, and provide a potential therapeutic strategy by targeting GATA1 and p2x7r locus binding.

Higher healthy lifestyle scores are associated with greater bone mineral density in middle-aged and elderly Chinese adults.

This study examined the association between healthy lifestyle score (HLS), which contained 7 items (smoking, BMI, physical activity, diet, alcohol, sleep and anxiety) and BMD. Results showed HLS was positively associated with BMD at all studied sites, suggesting that healthier lifestyle patterns might be beneficial to bone health. PURPOSE: Previous studies have reported favourable associations of individual healthy lifestyle factors with bone mineral density (BMD), but limited evidence showed the relationship of a combined healthy lifestyle score (HLS) with BMD. This study examined the association between the HLS and BMD. METHODS: This community-based cross-sectional study included 3051 participants aged 40-75 years. The HLS contained 7 items (smoking, BMI, physical activity, diet quality, alcohol intake, sleep and anxiety). BMD values of whole body (WB), lumbar spine 1-4 (L1-4), total hip (TH) and femur neck (FN) were measured using dual-energy X-ray absorptiometry. RESULTS: After adjusting for potential covariates, HLS was positively associated with BMD at all studied sites (P-trend < 0.01). The mean BMDs were 2.69% (WB), 5.62% (L1-4), 6.13% (TH) and 5.71% (FN) higher in participants with HLS of 6-7 points than in those with HLS of 0-2 points. The per 1 of 7 unit increase in the HLS was associated with increases of 7.63 (WB)-13.4 (TH) mg/cm2 BMD levels at all sites. These favourable associations tended to be more pronounced in men than in women. Among the 7 items, physical activity contributed most to the favourable associations, followed by BMI, non-smoking and diet; the other three items played little roles. Sensitivity analyses showed that the significant associations remained after excluding any one of the 7 components or excluding fracture subjects at all sites. CONCLUSION: Higher HLS was associated with greater BMD in middle-aged and elderly Chinese, suggesting that healthier lifestyle patterns might be beneficial to bone health.

Detection of the MYD88L265P and CXCR4S338X mutations by cell-free DNA in Waldenström macroglobulinemia.

We aimed to detect the MYD88L265P and CXCR4S338X mutations in cell-free DNA (cfDNA) in patients with Waldenström macroglobulinemia (WM). We collected peripheral blood and paired bone marrow aspirates from 27 WM patients (including 16 patients with newly diagnosed WM, 3 patients with WM in relapse and 8 patients with WM during treatment). cfDNA was extracted from peripheral blood using a QIAamp Circulating Nucleic Acid Kit. The MYD88L265P and CXCR4S338X mutations were detected by real-time allele-specific PCR (AS-PCR) in cfDNA and genomic DNA (gDNA) extracted from bone marrow aspirates. The sensitivity of real-time AS-PCR for detecting MYD88L265P in cfDNA was determined using a serial dilution of 10%, 2%, 0.4% and 0.08% MYD88L265P cfDNA in wild-type cfDNA. Among the 27 patients, MYD88L265P was detected in 88.9% of them in gDNA and in 85.2% of them in cfDNA, with a concordance rate of 96.3%. The concordance rates were 93.8%, 100% and 100% in patients with newly diagnosed WM, patients with WM in relapse and patients with WM during treatment, respectively. The sensitivity of real-time AS-PCR for detecting MYD88L265P in cfDNA was 0.4%. CXCR4S338X was detected in 6.3% of the 16 newly diagnosed WM patients in both gDNA and cfDNA, with a concordance rate of 100.0%. It is feasible to apply cfDNA to detect MYD88L265P and CXCR4S338X in WM patients with a high concordance rate.

The role of high serum CXCL13 level in Waldenström macroglobulinemia.

PURPOSE: To explore the value of elevated CXCL13 levels in Waldenström macroglobulinemia (WM). METHODS: We collected serum samples from 41 patients and bone marrow tissues from 14 patients with newly diagnosed symptomatic WM. Serum and bone marrow samples from patients with other indolent B-cell lymphomas and MGUS were also collected for comparison. Serum CXCL13 levels were measured by enzyme-linked immunosorbent assay, and bone marrow tissues were examined by immunohistochemistry. RESULTS: The median serum level of CXCL13 in patients with symptomatic WM was 2483.0 (range 36.8-5644.0) pg/ml, which was significantly higher than in patients with other indolent B-cell lymphomas with monoclonal IgM (median 380.9 pg/ml, range 23.8-5518.0 pg/ml) (p = 0.01). Serum CXCL13 >3250 pg/ml and serum M-protein >38 g/l diagnosed WM with a sensitivity of 98% and specificity of 85%. Serum CXCL13 was strongly correlated with hemoglobin levels (ρ=-0.46, p = 0.002), serum M-protein (ρ=0.47, p = 0.002), and IgM levels (ρ=0.30, p = 0.05) in patients with symptomatic WM. Immunohistochemistry analysis indicated that CXCL13 and activated mast cell levels were also higher in the bone marrow of WM patients compared to patients with IgM-MGUS or other indolent B-cell lymphomas with monoclonal IgM. CONCLUSIONS: Serum CXCL13 levels were significantly elevated in patients with WM and correlated with tumor load. Detection of serum CXCL13 may therefore be helpful in the differential diagnosis of WM.

Disparities in Diabetes Prevalence Among Native Hawaiians/Other Pacific Islanders and Asians in Hawai'i.

INTRODUCTION: The prevalence of diabetes varies widely among racial/ethnic groups in Hawai'i. How prevalence varies by age for Asian subgroups and Native Hawaiian/Other Pacific Islanders (NHOPIs) is understudied. We examined diabetes prevalence by age and race/ethnicity and assessed how socioeconomic status and lifestyle behaviors affected prevalence among Japanese, Filipino, Chinese, NHOPI, and white populations in Hawai'i. METHODS: We studied 18,200 subjects aged 18 or older from the Hawai'i Behavioral Risk Factor Surveillance System. We performed Poisson regression analyses to examine the prevalence of diabetes by race/ethnicity, age, sex, marital status, education, income, health care coverage, obesity, smoking and drinking status, physical activity, and fruit and vegetable consumption and examined the interactions of these factors with age and race/ethnicity. RESULTS: We found disparities in diabetes prevalence among respondents aged 35 to 44 and among Asians and NHOPIs, and disparities increased with age. NHOPIs and Filipinos had the highest prevalence of diabetes after controlling for other demographic factors and lifestyle variables. Japanese adults were less likely than NHOPIs and Filipinos to have diabetes; however, whites had the lowest prevalence. Income, physical activity, and obesity were the strongest predictors of diabetes. CONCLUSION: NHOPIs and Filipinos have higher rates of diabetes compared with other races/ethnicities in Hawai'i. More research is needed to reduce diabetes disparities among NHOPI and Filipino populations in Hawai'i. This study also shows the importance of conducting age-specific analyses of racial/ethnic-subgroups for health disparities.

Effects of berberine on tumor growth and intestinal permeability in HCT116 tumor-bearing mice using polyamines as targets.

The prognosis of colorectal cancer (CRC) is seriously affected by high intestinal mucosal permeability accompanied by increasing tumor load. Berberine, a natural plant-derived product, can protect the intestinal mucosal barrier and suppress tumor growth, but its effects on the intestinal mucosal barrier dysfunction of CRC have not yet been evaluated. Herein, we assessed the effects of berberine on the intestinal mucosal permeability of HCT116 tumor-bearing mice and the underlying mechanism. Berberine (6.25, 12.5, 25 mg/kg) was administered to tumor-bearing mice for 3 weeks by intraperitoneal injection, and saline was given to controls and models. Compared with the control group, tumor-bearing mice had increased intestinal mucosal permeability in the third week. Meanwhile, the body weight decreased by 4%-7%, the concentration of D-lactic acid in plasma increased, and the expressions of ZO1 and Occludin were down-regulated. The intestinal mucosa was impaired. Compared with the model group, berberine inhibited tumor growth in a dose-dependent manner (6.25, 12.5, 25 mg/kg), reduced the permeability of intestinal mucosa, and alleviated intestinal mucosal damage. HPLC showed that berberine decreased the content of polyamines in tumor tissue, whereas increased that in intestinal mucosa tissue. Western blot showed that berberine inhibited the expressions of ODC, C-MYC and HIF-1α, but up-regulated those of OAZ1 and SSAT. In short, berberine may exert antitumor effects by suppressing tumor growth and elevating the intestinal mucosal permeability.

Aspirin Increases the Risk of Nondiagnostic Yield of Fine-Needle Aspiration and Biopsy of Thyroid Nodules.

BACKGROUND: The link between the diagnostic yield of thyroid fine-needle aspiration and biopsy (FNAB) in patients taking antithrombotic or anticoagulant medications (AT/AC) remains poorly characterized. OBJECTIVES: We studied the risk of obtaining a nondiagnostic sample with ultrasound-guided thyroid FNAB in patients taking AT/AC medications. METHODS: This is a retrospective cohort study using medical rec-ords of 556 patients who underwent thyroid FNAB. All cytology samples were reported using the Bethesda classification. For patients with a nondiagnostic cytology, logistic regression was used to calculate OR for patients taking AT/AC medications. Multivariate regression was used to adjust for potential confounding variables including age, cystic ultrasound features, presence of eggshell calcifications, number of passes performed, cystic aspirate on FNAB, and position of the nodule. RESULTS: Out of 556 patients, cytology results were available for 547 patients. Of these, 46 subjects were taking aspirin and 1 was on warfarin. Among the entire cohort, 17.5% of the subjects had a nondiagnostic cytology. Among the patients on AT/AC medications, 34% had a nondiagnostic result compared to 16% for those not taking them (OR = 2.70, p = 0.003). The subgroup of patients taking aspirin had similarly higher odds of a nondiagnostic cytology (OR = 2.78, p = 0.002). These differences remained statistically significant after multivariate adjustment. CONCLUSIONS: This is the first study to demonstrate a 3-fold independently greater risk of a nondiagnostic FNAB cytology in patients taking aspirin. Our results highlight the importance of evaluating the need for continuation of aspirin in patients undergoing thyroid FNAB as this may impact the diagnostic yield of the procedure.

Ultrasound-guided needle release of the transverse carpal ligament with and without corticosteroid injection for the treatment of carpal tunnel syndrome.

BACKGROUND: To compare the clinical effectiveness of ultrasound-guided needle release of the transverse carpal ligament (TCL) with and without corticosteroid injection in carpal tunnel syndrome (CTS). METHODS: From June 2016 to June 2017, 49 CTS patients (50 wrists) were included in this study. Twenty-five wrists were treated with ultrasound-guided needle release of the TCL plus corticosteroid injection (group A), and 25 wrists were treated with single ultrasound-guided needle release of the TCL (group B). The following parameters were assessed and compared including postprocedure results according to relief of symptoms, ultrasound parameters (cross-sectional area of the median nerve at the levels of pisiform, flattening ratio of median nerve at the levels of the hamate bone, and the thicknesses of TCL on the cross-section at the level of the hamate bone), and electrophysiological parameters (distal motor latency and sensory conduction velocity). RESULTS: Group A had higher overall excellent and good rate 3 months after the procedure than group B (84 vs 52%, P < 0.05). There were significant differences regarding the above ultrasonic and electrophysiological parameters between the baseline and postprocedure values in both groups (all P < 0.05). There were significant differences regarding the postprocedure values of above ultrasonic and electrophysiological parameters between the two groups (all P < 0.05). No complications such as infection or tendon rupture were noted. No procedures were converted to the open release. CONCLUSIONS: Both techniques are effective in treating CTS. Ultrasound-guided needle release of the TCL with corticosteroid injection had better treatment benefits than single ultrasound-guided needle release of the TCL in treating CTS.

Model-based recursive partitioning to identify risk clusters for metabolic syndrome and its components: findings from the International Mobility in Aging Study.

OBJECTIVE: Conceptual models underpinning much epidemiological research on ageing acknowledge that environmental, social and biological systems interact to influence health outcomes. Recursive partitioning is a data-driven approach that allows for concurrent exploration of distinct mixtures, or clusters, of individuals that have a particular outcome. Our aim is to use recursive partitioning to examine risk clusters for metabolic syndrome (MetS) and its components, in order to identify vulnerable populations. STUDY DESIGN: Cross-sectional analysis of baseline data from a prospective longitudinal cohort called the International Mobility in Aging Study (IMIAS). SETTING: IMIAS includes sites from three middle-income countries-Tirana (Albania), Natal (Brazil) and Manizales (Colombia)-and two from Canada-Kingston (Ontario) and Saint-Hyacinthe (Quebec). PARTICIPANTS: Community-dwelling male and female adults, aged 64-75 years (n=2002). PRIMARY AND SECONDARY OUTCOME MEASURES: We apply recursive partitioning to investigate social and behavioural risk factors for MetS and its components. Model-based recursive partitioning (MOB) was used to cluster participants into age-adjusted risk groups based on variabilities in: study site, sex, education, living arrangements, childhood adversities, adult occupation, current employment status, income, perceived income sufficiency, smoking status and weekly minutes of physical activity. RESULTS: 43% of participants had MetS. Using MOB, the primary partitioning variable was participant sex. Among women from middle-incomes sites, the predicted proportion with MetS ranged from 58% to 68%. Canadian women with limited physical activity had elevated predicted proportions of MetS (49%, 95% CI 39% to 58%). Among men, MetS ranged from 26% to 41% depending on childhood social adversity and education. Clustering for MetS components differed from the syndrome and across components. Study site was a primary partitioning variable for all components except HDL cholesterol. Sex was important for most components. CONCLUSION: MOB is a promising technique for identifying disease risk clusters (eg, vulnerable populations) in modestly sized samples.

Venous and arterial base deficits: do these agree in occult shock and in the elderly? A Bland-Altman analysis.

BACKGROUND: Trauma centers are increasingly advocating the replacement of arterial blood gas measurements with venous blood gas measurements for simplification of base deficit (BD) determination. These values have never been demonstrated to agree in important trauma populations, such as for patients in occult shock (OS) or the elderly. The goal of this study was to investigate the level of agreement between venous and arterial BDs from blood gases in critically ill or injured patients, specifically in OS and the elderly. METHODS: This is a retrospective, consecutive, cohort study using matched pairs of venous and arterial blood gases from patients admitted to the Trauma and Neurosurgery Intensive Care Unit in a Level I trauma center in Toronto, Ontario, Canada. Agreement between near simultaneous arterial and venous BD was calculated using the Bland-Altman method. McNemar's test was used for differences in BDs in the presence or absence of OS and in elderly patients. RESULTS: BDs for 466 arterial and venous samples from 72 patients were compared pairwise. There was no significant difference between samples (p = 0.88). Ninety-eight percent of samples were within 3.0 mmol/L of each other. No significant differences were detected between venous and arterial BD in the presence of OS or in the elderly (p = 0.72 and p = 0.25, respectively). CONCLUSION: Arterial and venous BDs agree, including in the presence of OS and in the elderly. Consideration may be given to venous sampling both in the intensive care unit or in other areas of care, such as the trauma bay. LEVEL OF EVIDENCE: Diagnostic study, level III.

A novel viral SOCS from infectious spleen and kidney necrosis virus: interacts with Jak1 and inhibits IFN-α induced Stat1/3 activation.

Interferon (IFN)-induced Janus kinase (Jak)/signal transducer and activator of transcription (Stat) pathway is important in controlling immune responses and is negatively response-regulated by the suppressor of cytokine signaling (SOCS) proteins. However, several viruses have developed various strategies to inhibit this pathway to circumvent the anti-viral immunity of the host. The infectious spleen and kidney necrosis virus (ISKNV) is the type species of the genus Megalocytivirus in the family Iridoviridae and a causative agent of epizootics in fish. ISKNV ORF103R encodes a predicted viral SOCS (vSOCS) with high homology to the vertebrate SOCS1, but lacks a SOCS-box domain. Interestingly, vSOCS only exists in the genus Megalocytivirus. ISKNV-vSOCS can block the IFN-α-induced Jak/Stat pathway in HepG2 cells. Over-expression of ISKNV-vSOCS inhibited the activities of IFN-stimulated response element (ISRE) promoter; however, the inhibitions by ISKNV-vSOCS were dose-dependent. ISKNV-vSOCS interacted with Jak1 protein and inhibited its tyrosine kinase activity in vitro. ISKNV-vSOCS also impaired the phosphorylation of Stat1 and Stat3 proteins and suppressed their activations. The point mutations (F18D, S66A, S85A, and R64K) of ISKNV-vSOCS significantly impaired the inhibition of IFN-α-induced ISRE-promoter activation. In conclusion, vSOCS inhibits IFN-α-induced Stat1/Stat3 signaling, suggesting that Megalocytivirus has developed a novel strategy to evade IFN anti-viral immunity via vSOCS protein.

Entry of tiger frog virus (an Iridovirus) into HepG2 cells via a pH-dependent, atypical, caveola-mediated endocytosis pathway.

Tiger frog virus (TFV), in the genus Ranavirus of the family Iridoviridae, causes high mortality of cultured tiger frog tadpoles in China. To explore the cellular entry mechanism of TFV, HepG2 cells were treated with drugs that inhibit the main endocytic pathways. We observed that TFV entry was inhibited by NH(4)Cl, chloroquine, and bafilomycin, which can all elevate the pH of acidic organelles. In contrast, TFV entry was not influenced by chlorpromazine or overexpression of a dominant-negative form of Esp15, which inhibit the assembly of clathrin-coated pits. These results suggested that TFV entry was not associated with clathrin-mediated endocytosis, but was related to the pH of acidic organelles. Subsequently, we found that endocytosis of TFV was dependent on membrane cholesterol and was inhibited by the caveolin-1 scaffolding domain peptide. Dynamin and actin were also required for TFV entry. In addition, TFV virions colocalized with the cholera toxin subunit B, indicating that TFV enters as caveola-internalized cargo into the Golgi complex. Taken together, our results demonstrated that TFV entry occurs by caveola-mediated endocytosis with a pH-dependent step. This atypical caveola-mediated endocytosis is different from the clathrin-mediated endocytosis of frog virus 3 (FV3) by BHK cells, which has been recognized as a model for iridoviruses. Thus, our work may help further the understanding of the initial steps of iridovirus infection in lower vertebrates.

Involvement of caveolin-1 in the Jak-Stat signaling pathway and infectious spleen and kidney necrosis virus infection in mandarin fish (Siniperca chuatsi).

Caveolae, the major source of caveolin-1 protein, are specialized invaginated microdomains of the plasma membrane that act as organizing centers for signaling molecules in the immune system. In the present study, we report the cloning and characterization of caveolin-1 (mCav-1) from mandarin fish (Siniperca chuatsi) and study on the roles of mCav-1 in the fish Jak-Stat signaling pathway and in virus infection. The cDNA sequence of mCav-1 was 707bp in size, encoding a protein of 181 amino acids, which was different from the mammalian protein (178 amino acids). The deduced amino acid sequence of mCav-1 shared similar architecture with vertebrate caveolin-1 proteins, but mCav-1 lacked a phosphorylation site (y14). The major subcellular location of mCav-1 was in the caveolae, where the protein appeared to have major functions. Real-time PCR revealed that the expression of the mandarin fish Mx, IRF-1, SOCS1, and SOCS3 genes involved in the poly(I:C)-induced Jak-Stat signaling pathway was impaired by the mCav-1 scaffolding domain peptide (mSDP). In mandarin fish fry (MFF-1) cells, the protein levels of mCav-1 were markedly up-regulated at 12 and 24h post-infection with ISKNV (infectious spleen and kidney necrosis virus). In addition, ISKNV entry into MFF-1 cells was significantly inhibited by mSDP, and the inhibition was dose-dependent. Thus, ISKNV infection was apparently associated with mCav-1 protein and may utilize the caveolae-related endocytosis pathway. The findings reported here further our understanding of the function of caveolin-1 in the complex signal transduction network in fish immune systems and in the cellular entry mechanism of iridoviruses.

[Effect of occupational stress on hypertension].

OBJECTIVE: To explore the effect of occupational stress on hypertension. METHODS: 498 workers whose accumulative length of service was more than two years were investigated with questionnaire by method of cluster sampling from a thermal power plant in Henan province in China; 446 respondents returned qualified questionnaire including 281 male and 165 female Han People. After the patients with secondary hypertension, diabetes patients, and patients with liver or kidney disease were excluded, 84 workers (58 males and 26 females) were diagnosed as hypertension. The occupational stressors, personalities, buffering factors and occupational strain were measured by using the Job Demand-control Model, the Effort-reward Imbalance Model questionnaires and Occupational Stress Measurement Scale. Main risk factors for the development of hypertension such as heredity, body mass index, high salt diet, alcohol use, smoking habit and lack of physical activity were investigated. 498 whole blood samples were collected from workers in field epidemiologic survey. All of the samples were detected TG, CHO and FPG by common biochemistry methods. Multivariate logistic regression analysis were used to determine the relationship between occupational stressors and prevalence rate of hypertension. The difference of morbidity of hypertension between different stress level subjects was analyzed by chi2 test. RESULTS: (1) Logistic regression analysis of the hypertension by all occupational stressors and risk factors of hypertension indicated that not only some common factors such as parents' hypertensive history, BMI, alcohol use and TG, but also responsibility for person, work locus of control and social support were significantly correlated with elevated risks of hypertension. (2) Logistic regression analysis of the hypertension by main dimensions of effort-reward imbalance model and risk factors of hypertension indicated that parents' hypertensive history, BMI, alcohol use, TG, and effort were significantly correlated with elevated risks of hypertension. Logistic regression analysis indicated the risk of hypertension had an effect on the FRI and effort (OR was 1.71 and 2.43 respectively). (3) For the job strain model, results indicated that parents' hypertensive history, UMI, alcohol use, TG, work locus of control and social support were significantly correlated with elevated risks of hypertension. But the main dimensions of job strain model (job demands and decision latitude) didn't enter regression equation. (4) The difference of prevalence of hypertension between high- and low stress level groups in male was statistically significant (OR = 3.13, P < 0.01), but the case was not the same in female (P > 0.05). CONCLUSIONS: Occupational stress might be risk factor of hypertension; The predictive power of effort-reward imbalance model for the development of hypertension would be larger than that of job strain model.

[Study on the effect of heluoshugan capsule on liver fibrosis induced by Schistosoma japonicum infection in mice].

OBJECTIVE: To investigate the effect of Chinese traditional medicine heluoshugan capsule on liver fibrosis induced by Schistosoma japonicum infection in mice. METHODS: Liver fibrosis in mice was established by Schistosoma japonicum infection in 6 weeks. Suspension of heluoshugan prepared with normal saline was given orally to the mice, 2 capsules for 20 mice daily for 8 weeks. The level of vascular endothelial growth factor (VEGF), focal adhesion kinase (FAK) and type I, III collagen in liver tissue were detected by immuno-histochemistry. RESULTS: The results showed that heluoshugan improved the pathological change of the liver tissue, decreased the level of type I, III collagen, especially type III collagen (P < 0.01). The level of VEGF and FAK expression was inhibited after the administration of heluoshugan, though the level usually increased in liver fibrosis due to the infection. CONCLUSIONS: The result suggests that heluoshugan capsule might have therapeutic effect on liver fibrosis induced by the infection of Schistosoma japonicum in mice, by inhibiting the activation of hepatic stellate cells and the pathological change of liver blood vessel.