VG161 activates systemic antitumor immunity in pancreatic cancer models as a novel oncolytic herpesvirus expressing multiple immunomodulatory transgenes.

The VG161 represents the first recombinant oncolytic herpes simplex virus type 1 carrying multiple synergistic antitumor immuno-modulating factors. Here, we report its antitumor mechanisms and thus provide firm theoretical foundation for the upcoming clinical application in pancreatic cancer. Generally, the VG161-mediated antitumor outcomes were analyzed by a collaboration of techniques, namely the single-cell sequencing, airflow-assisted desorption electrospray ionization-mass spectrometry imaging (AFADSI-MSI) and nanostring techniques. In vitro, the efficacy of VG161 together with immune checkpoint inhibitors (ICIs) has been successfully shown to grant a long-term antitumor effect by altering tumor immunity and remodeling tumor microenvironment (TME) metabolisms. Cellular functional pathways and cell subtypes detected from patient samples before and after the treatment had undergone distinctive changes including upregulated CD8+ T and natural killer cells. More importantly, significant antitumor signals have emerged since the administration of VG161 injection. In conclusion, VG161 can systematically activate acquired and innate immunity in pancreatic models, as well as improve the tumor immune microenvironment, indicative of strong antitumor potential. The more robusting antitumor outcome for VG161 monotherapy or in combination with other therapies on pancreatic cancer is worth of being explored in further clinical trials.

A Cas12a-based fluorescent microfluidic system for rapid on-site human papillomavirus diagnostics.

Persistent infection with human papillomavirus (HPV) is the leading cause of cervical cancer, and early diagnosis is crucial for clinical management. However, the easy and rapid on-site diagnostic for HPV genotyping remains challenging. Here, we develop a Cas12a-based fluorescent microfluidic detection system for diagnosing six HPV subtypes (HPV6, HPV11, HPV16, HPV18, HPV31, and HPV33). A panel of crRNAs and recombinase polymerase amplification (RPA) primers targeting the HPV L1 gene was screened for sensitive and specific detection. Furthermore, a one-pot RPA reaction was developed to amplify the six HPV subtypes without cross-reactivity. For on-site detection, we integrated the RPA-Cas12a detection into a microfluidic device, enabling the detection of processed clinical samples within 35 minutes. The assay was validated using 112 clinical swab samples and obtained consistent results with the qPCR assay, with a concordance rate of 99.1%. Overall, our diagnostic method offers a rapid, sensitive, and easy-to-use on-site assay for detecting HPV genotypes and holds promise for improving cervical cancer screening and prevention. KEY POINTS: • The Cas12a-based fluorescent microfluidic detection system for the diagnosis of six HPV subtypes. • A one-pot RPA reaction for amplifying the six HPV subtypes without cross-reactivity. • The RPA-Cas12a-microfluidic system provides results within 35 minutes for on-site detection.

Thrombocytopenia in patients with hepatitis B virus-related chronic hepatitis: evaluation of the immature platelet fraction.

The immature platelet fraction (IPF) measures the number of reticulated platelets in peripheral blood, and can be used to help determine if thrombocytopenia is secondary to low-platelet production or increased platelet turnover. The aim of this study was to determine whether abnormalities in the IPF were associated with thrombocytopenia in patients with hepatitis B virus-related chronic hepatitis (CHB). One hundred fifty-six patients with chronic hepatitis B, including 80 thrombocytopenia, 76 without thrombocytopenia, and 48 healthy controls were enrolled in the study. The IPF percentages (IPF%) were measured using a XE-2100 multiparameter automatic hematology analyzer. We demonstrated that in the thrombocytopenic group, the IPF% was significantly increased compared with that in healthy controls and the non-thrombocytopenic group (both p < 0.001). Multivariate analysis demonstrated that IPF%, splenomegaly, and the model for end-stage liver disease score were independent predictors for thrombocytopenia (both p < 0.001). High IPF% during the course of thrombocytopenia suggests that platelet destruction/sequestration due to hypersplenism is a major factor contributing to thrombocytopenia in patients with CHB.

Evaluation of an automated microplate technique in the Galileo system for ABO and Rh(D) blood grouping.

BACKGROUND: A number of automated devices for pretransfusion testing have recently become available. This study evaluated the Immucor Galileo System, a fully automated device based on the microplate hemagglutination technique for ABO/Rh (D) determinations. METHODS: Routine ABO/Rh typing tests were performed on 13,045 samples using the Immucor automated instruments. Manual tube method was used to resolve ABO forward and reverse grouping discrepancies. D-negative test results were investigated and confirmed manually by the indirect antiglobulin test (IAT). RESULTS: The system rejected 70 tests for sample inadequacy. 87 samples were read as "No-type-determined" due to forward and reverse grouping discrepancies. 25 tests gave these results because of sample hemolysis. After further tests, we found 34 tests were caused by weakened RBC antibodies, 5 tests were attributable to weak A and/or B antigens, 4 tests were due to mixed-field reactions, and 8 tests had high titer cold agglutinin with blood qualifications which react only at temperatures below 34 degrees C. In the remaining 11 cases, irregular RBC antibodies were identified in 9 samples (seven anti-M and two anti-P) and two subgroups were identified in 2 samples (one A1 and one A2) by a reference laboratory. As for D typing, 2 weak D+ samples missed by automated systems gave negative results, but weak-positive reactions were observed in the IAT. CONCLUSIONS: The Immucor Galileo System is reliable and suited for ABO and D blood groups, some reasons may cause a discrepancy in ABO/D typing using a fully automated system. It is suggested that standardization of sample collection may improve the performance of the fully automated system.

The mean platelet volume is a potential biomarker for cirrhosis in chronic hepatitis B virus infected patients.

BACKGROUND/AIMS: In recent years, a great interest has been dedicated to the development of noninvasive predictive models to substitute liver biopsy for cirrhosis assessment and follow-up. Our aim was to provide a simple marker for predicting liver cirrhosis in chronic hepatitis B virus (HBV)-infected patients based on routinely available clinical parameters. METHODOLOGY: Three hundred and fifty-four patients with chronic hepatitis B, including 144 with cirrhosis, 210 without cirrhosis, and 68 healthy controls were enrolled in the study. During the study, a blood sample was collected from all cases to examine liver function, renal function, international normalized ratio (INR) and routine hematological testing. RESULTS: We demonstrated that in the cirrhosis group, mean platelet volume (MPV) values were significantly increased compared with healthy controls and non-cirrhosis group. Multivariate analysis demonstrated that MPV, total bilirubin (TB) and INR were independent predictors for cirrhosis (both P < 0.01). CONCLUSIONS: MPV is a simple and non-invasive routine laboratory parameters and elevated MPV level might be an independent predictor for cirrhosis in patients with chronic HBV infection.

Changes in peripheral T-lymphocyte subsets in acute-on-chronic liver failure patients with artificial liver support system.

BACKGROUND/AIMS: HBV-related acute-on-chronic liver failure (AoCLF) is associated with a high mortality rate. An artificial liver support system (ALSS) is a newly emerging therapeutic option, which can be implemented in routine patient care. In order to determine if any pretreatment immunological parameter could be an indicator to evaluate immune states for the outcome of the AoCLF patients, we analyzed the relationship between the level of T-lymphocyte subsets (CD4+, CD8+, CD4/CD8 ratio) and its therapeutic effect and prognosis. METHODOLOGY: Sixty-three patients with AoCLF were enrolled in 2 groups (ALSS plus routine-care, n=29 and routine-care only, n=34). In the ALSS group, there were 17 survivors (17/29), while in the routine-care group there were 11 survivors (11/34), both after 30 days of treatment. Twenty-three healthy donors were used as the control group. The number of CD4 and CD8 T cells was detected by flow cytometry and the ratio of CD4/ CD8 was calculated on admission and on days 7, 14, 21 and 30, during hospitalization. RESULTS: More dramatic increased CD4/CD8 ratio in the ALSS survivors (form 0.92±0.18 to 1.26±0.24, p<0.01) than the medical survivors (form 0.86±0.16 to 1.09±0.16, p<0.05) after 30 days of treatment. A declined tendency was observed in nonsurvivors. CONCLUSIONS: T-lymphocyte subsets may be important in the pathogenesis of the AoCLF and ALSS may represent a reliable hepatic support device for Ao- CLF.

Tranilast attenuates lipopolysaccharide­induced lung injury via the CXCR4/JAK2/STAT3 signaling pathway.

It has been reported that the expression of C­X­C motif chemokine receptor 4 (CXCR4) is increased in patients with lung injury, while CXCR4 downregulation can improve sepsis­induced lung injury. Previous studies have shown that tranilast can inhibit CXCR4 mRNA expression. Therefore, the present study aimed to investigate whether tranilast could protect against lipopolysaccharide (LPS)­induced lung injury via the CXCR4/Janus kinase 2 (JAK2)/STAT3 signaling pathway. A Cell Counting Kit­8 assay was performed to evaluate the effect of different concentrations of tranilast on the viability of LPS­induced BEAS­2B cells. The mRNA and protein expression levels of the inflammatory factors, TNFα, IL­1ß, IL­6, cytochrome c oxidase subunit II and inducible nitric oxide synthase were detected using reverse transcription­quantitative PCR and western blot analysis, respectively. In addition, the cell apoptosis rate and the expression levels of apoptosis­related proteins were analyzed using a TUNEL staining assay and western blot analysis, respectively. The expression levels of the CXCR4/JAK2/STAT3 signaling pathway­related proteins were also determined using western blot analysis. Furthermore, the effects of tranilast on cell viability, inflammation and apoptosis were also evaluated in LPS­stimulated BEAS­2B cells following CXCR4 overexpression, which were pre­treated with tranilast. The results demonstrated that tranilast could alleviate LPS­induced cell viability, the secretion of inflammatory cytokines and cell apoptosis. In addition, cell treatment with tranilast inhibited the expression of CXCR4/JAK2/STAT3 signaling­related proteins in LPS­induced BEAS­2B cells. Following CXCR4 overexpression, the alleviating effect of tranilast on cell viability, inflammatory response and apoptosis was notably attenuated. Overall, the current study suggested that tranilast could attenuate LPS­induced lung injury via the CXCR4/JAK2/STAT3 signaling pathway, suggesting that tranilast could be considered as a promising agent for treating sepsis­induced acute lung injury.

Sustainable Treatment and Resource Recovery of Anion Exchange Spent Brine by Pilot-Scale Electrodialysis and Ultrafiltration.

The anion exchange (AIX) spent brine, generated during the NDMP-3 resin regeneration process, highly loaded with organic substances mainly humic substances (HSs) and salts (mainly NaCl) remains an environmental concern. In this study, pilot-scale electro dialysis (ED) and ultrafiltration (UF) hybrid technologies were first used to recover NaCl solution as a resin regeneration agent and HSs, which could be utilized as a vital ingredient of organic fertilizer, from the AIX spent brine. Recovered ≈ 15% w/w NaCl solution obtained by two-stage pilot-scale ED can be used to regenerate saturated NDMP-3 anion exchange resins; the regeneration−readsorption performance of NDMP-3 resins was equivalent to that of fresh ≈ 15% w/w NaCl solution. The two-stage dilute solution with low-salt content (0.49% w/w) was further concentrated by pilot-scale UF, so that the HS content in the retentate solution was >30 g/L, which meets the HS content required for water-soluble organic fertilizers. The HS liquid fertilizer could significantly stimulate the growth of green vegetables with no phytotoxicity, mainly due to special properties of HSs. These results demonstrate that ED + UF hybrid technologies can be a promising approach for the sustainable treatment and resource recovery of AIX spent brine.

Intravenous leiomyomatosis of the uterus with extension to the right heart.

A 42-year-old woman admitted with debilitation and engorgement both lower extremities. Transthoracic two-dimensional echocardiography, abdominal ultrasound and computerized tomography revealed a lobulated pelvic mass, a mass within right internal iliac vein, both common iliac vein, as well as the inferior vena cava, extending into the right atrium. In addition, echocardiography and abdominal ultrasound showed the tumor of right atrium and inferior vena cave has no stalk and has well-demarcated borders with the wall of right atrium and inferior vena cave. Hence, the presumptive diagnosis of IVL was made by echocardiography and abdominal ultrasound and the presumptive diagnosis of sarcoma with invasion in right internal iliac vein, both common iliac vein, the inferior vena cava, as well as the right atrium was made by multi-detector-row computerized tomography. The patient underwent a one-stage combined multidisciplinary thoraco-abdominal operation under general anaesthetic. Subsequently the pathologic report confirmed IVL.

Reduction in rectal doses of pelvic radiotherapy with excessive rectal gas extraction: a single-institution case report.

During radiotherapy for pelvic tumors, there might be insufficient radiation doses delivered that result from concerns of immediately adjacent critical organs, most notebly the rectum. Several measures have been proposed to reduce the rectal doses, such as use of a radioprotective spacer to shift the rectum away from the lesion or an endorectal balloon to reduce the internal target volume (ITV). The removal of rectal gas is supposed to achieve the similar effect by reducing the rectal volume and ITV. This is because the distance of the rectum to lesions can be increased with decreasing rectal volumes, which also makes it easy for physicists to avoid the rectum during field design. Besides, a reduced ITV can be obtained due to stability in rectal volume, as with use of an endorectal balloon. The procedure is most suitable for patients receiving stereotactic body radiotherapy (SBRT) and high dose rate brachytherapy (HDR-BR) because of limited treatment sessions. In this study, we showed that the rectal dose decreased significantly with rectal gas extraction, as manifested by dose parameters of D2 cm3 and D0.1 cm3 (the doses delivered to the most exposed 2 cm, 3 and 0.1 cm3 of the rectum, respectively). This technique is especially benefitable for candidates with excessive rectal gas fillings.

microRNA-15a-5p participates in sepsis by regulating the inflammatory response of macrophages and targeting TNIP2.

The mortality rate for patients experiencing sepsis is decreasing; however, an effective therapeutic strategy requires further investigation. Increasing evidence has supported the idea that dysregulated microRNAs (miR) participate in the development of sepsis. Meanwhile, macrophages are crucial players in various inflammatory responses and diseases. The objective of the current study was to investigate the associated molecular mechanisms of action of miR-15a-5p on inflammatory responses in lipopolysaccharide (LPS)-stimulated mouse macrophages and the macrophage cell line RAW264.7. RAW264.7 macrophages were stimulated with LPS for 4 h, and ELISAs were subsequently used to measure the expression levels of pro-inflammatory cytokines, including tumor necrosis factor (TNF)-α, interleukin (IL)-1ß and IL-6, in RAW264.7 macrophages. The expression levels of miR-15a-5p in RAW264.7 macrophages were detected after the stimulation of LPS using reverse transcription quantitative-PCR. The results indicated that the IL-1ß, IL-6, TNF-α and miR-15a-5p levels were significantly increased compared with the control group. The Target gene prediction database (TargetScan) and dual-luciferase reporter assays were subsequently employed, and TNF-α induced protein 3-interacting protein 2 (TNIP2) was confirmed as a direct target for miR-15a-5p. Additionally, it was found that the TNIP2 expression levels were decreased in RAW264.7 macrophages following LPS treatment compared with controls. The present study also examined the effects of miR-15a-5p inhibitor on inflammatory cytokine expression levels and the activation of the NF-κ signaling pathway. These results demonstrated that miR-15a-5p inhibitor reduced the secretion of inflammatory cytokines and inhibited NF-κ pathway activation by targeting TNIP2. This may be associated with the progression of sepsis. Meanwhile, a LPS-induced mouse model of sepsis was established to examine the regulation of TNIP2 and miR-15a-5p during inflammation. In the animal model, miR-15a-5p inhibitor significantly suppressed the secretion of inflammatory factors. The levels of creatin, blood urea nitrogen, aspartate aminotransferase and alanine aminotransferase in the serum of LPS-treated mice were also found to be decreased in the miR-15a-5p inhibitor treatment group, while the protective effects of miR-15a-5p inhibitor on sepsis were eliminated by TNIP2-small interfering RNA combination therapy. In conclusion, the present findings indicated that miR-15a-5p may be involved in the inflammatory process during sepsis by activating the NF-κ pathway and targeting TNIP2. This suggests that miR-15a-5p inhibitor may be a novel anti-inflammatory agent and therapeutic strategy for sepsis.

Tunable odd-frequency triplet pairing states and skyrmion modes in chiral p-wave superconductor.

Bogliubov-de Gennes equations are solved self-consistently to investigate the properties of bound states in chiral p-wave superconductive disks. It shows that either an s-wave or the mixed d- and s-wave state with odd-frequency and spin-triplet symmetry is induced at the vortex core, depending both on the chirality of the pairing states and on the vortex topology. It is also found that the odd-frequency triplet even parity (OTE) bound state can be manipulated with a local non-magnetic potential. Interestingly, with an appropriate potential amplitude, the zero-energy OTE bound state can be stabilized at a distance from the vortex core and from the local potential. Possible existences of the Majorana fermion modes are expected if the particle-hole symmetry property is applied to the zero-energy OTE bound state. Moreover, skyrmion modes with an integer topological charge have been found to exist.

Serum C-reactive protein predicts early mortality in hospitalized patients with HBV-related decompensated cirrhosis.

The serum C-reactive protein (CRP) is an inflammatory marker. The aim of the present study was to elucidate whether CRP could serve as a potential surrogate marker for 30-day mortality in hospitalized patients with HBV-related decompensated cirrhosis (HBV-DeCi).This was a retrospective cohort study that included 140 patients with HBV-DeCi. All patients were followed up for 1-month. A panel of clinical and biochemical variables were analyzed for potential associations with outcomes using multiple regression models.The serum CRP was significantly higher in nonsurviving patients than in surviving patients. Multivariate analysis demonstrated that CRP levels (odds ratio: 1.047, P = 0.002) and the model for end-stage liver disease score (odds ratio: 1.370, P = 0.001) were independent predictors for mortality.Serum CRP is a simple marker that may serve as an additional predictor of 1-month mortality in hospitalized patients with HBV-DeCi.

Lymphocyte-monocyte ratio at admission predicts possible outcomes in patients with acute-on-chronic liver failure.

BACKGROUND: The lymphocyte-monocyte ratio (LMR) in the peripheral blood is suggested to be a potential biomarker for predicting the clinical outcomes of several diseases. We aimed to evaluate the relative efficiency of LMR for predicting 3-month mortality in patients with acute-on-chronic liver failure (AoCLF). PATIENTS AND METHODS: In this study, 74 chronic hepatitis B patients, 90 AoCLF patients, and 70 healthy controls were followed up for 4 months. The primary endpoint was 3-month in-hospital mortality. Hematological and virological parameters as well as liver biochemistry were determined using blood samples ordered upon admission. A panel of clinical and biochemical variables were analyzed for potential associations with outcomes using Cox proportional hazards and multiple regression models. RESULTS: A significantly lower LMR was detected in AoCLF patients than in healthy controls and chronic hepatitis B groups (both P=0.001). The LMR inversely correlated with model for end-stage liver disease scores, and a lower LMR was associated with increased 3-month mortality. Multivariate analysis suggested that both LMR and model for end-stage liver disease scores were independent predictors of 3-month mortality (P<0.01). CONCLUSION: A low LMR measured at admission is predictive of a poor prognosis in AoCLF patients.

Abnormal serum iron markers in chronic hepatitis B virus infection may be because of liver injury.

OBJECTIVE: In patients with chronic hepatitis B virus (HBV) infection, it is not known whether altered serum iron markers are directly because of the infection or the associated liver injury. We determined the serum iron status of patients with chronic HBV infection, and investigated whether it is HBV infection or HBV-related liver injury that likely causes abnormal serum iron markers in chronic HBV infection. MATERIALS AND METHODS: For a retrospective study, chronic HBV-infected patients (80 patients with cirrhosis and 76 patients without cirrhosis) and 58 healthy controls were enrolled. Serum alanine transaminase levels were measured to ascertain liver damage. Indicators of iron status included serum iron, ferritin, and transferrin. RESULTS: Compared with noncirrhotic patients and healthy controls, the serum transferrin of cirrhotic patients was lower and the serum iron and ferritin values were higher (P < 0.001, all). In cirrhotic patients, the serum iron and ferritin levels correlated positively with serum alanine transaminase levels and the transferrin levels were inversely related to both end-stage liver disease scores and iron levels (all P < 0.01). CONCLUSION: Serum iron markers tended to be aberrant in chronic HBV-infected patients with cirrhosis. The liver injury associated with HBV infection, but not chronic HBV infection directly, is likely the main cause for iron metabolism disorder.

Immature platelet fraction values predict recovery of platelet counts following liver transplantation.

BACKGROUND: Thrombocytopenia after orthotopic liver transplantation (OLT) is a well-recognized and prevalent early postoperative complication. In the current study, we assessed the clinical utility of the immature platelet fraction (expressed as a percent of total platelets, IPF %) to predict platelet count recovery following OLT. METHODS: We analyzed the changes in peripheral platelet counts, IPF values, and liver function before OLT, and 21 days after transplantation in 30 patients with liver cirrhosis. All patients were followed up for at least six months. RESULTS: The nadir in the peripheral platelet counts most commonly occurred on the 5th day after post-transplant. Platelets counts gradually rose again to exceed pre-transplant levels by the 3rd to 4th day after the IPF % reached its peak. CONCLUSION: The IPF % increased prior to the elevation of platelet counts in patients with OLT suggesting that the IPF % may reflect production of platelets. These findings suggest IPF% could be useful as a predictor of platelet recovery in patients with OLT.

Evaluation of mean platelet volume in patients with hepatitis B virus infection.

OBJECTIVE: Mean platelet volume (MPV), which is determined by a routine complete blood count, is a parameter that is usually overlooked by clinicians. The present study was designed to investigate the association between MPV and different disease states in patients with hepatitis B virus (HBV) infection. METHODS: A total of 120 patients, including 17 with acute hepatitis B (AHB), 62 with chronic hepatitis B (CHB), and 41 with chronic severe hepatitis B (CSHB), as well as 58 healthy controls (HCs) were enrolled in the study. At study entry, blood samples were collected from all subjects to examine liver function and renal function, determine the international normalized ratio and perform routine hematological tests. RESULTS: We demonstrated that MPV was significantly increased in CSHB and CHB patients compared with HCs and AHB patients (all P<0.05). Among the patient groups, the CSHB patients had the highest MPV. Increased MPV was clinically associated with severe liver disease. CONCLUSIONS: MPV is significantly increased in chronic HBV-infected patients and is associated with disease severity; thus, it may serve as an important biomarker.

Low Serum retinol-binding protein-4 levels in acute exacerbations of chronic obstructive pulmonary disease at intensive care unit admission is a predictor of mortality in elderly patients.

BACKGROUND: Acute exacerbations of chronic obstructive pulmonary disease (AECOPD) are thought to be associated with increased mortality in elderly patients. Low retinol-binding protein-4 (RBP4) is associated with a high risk of respiratory infections in the general population. Therefore, we hypothesized that low RBP4 levels are associated with an increased risk of AECOPD and can be used as a biomarker for AECOPD in elderly patients. METHODS: Enzyme-linked immunosorbent assays were used to assess RBP4 levels in elderly with AECOPD within the first 24 hours after intensive care unit admission. Forty-six elderly patients with stable COPD in outpatient clinics and 50 healthy elderly persons who had physical examinations as outpatients were controls. RESULTS: In AECOPD patients, RBP4 levels were lower than those in stable COPD patients and healthy controls (59.7 vs 91.2 and 113.6 mg/L, p < 0.001). RBP4 levels were decreased by 30.6% in non-survivors compared with survivors (51.5 vs 74.2 mg/L, p < 0.001). A higher Acute Physiology and Chronic Health Enquiry II (APACHE II) score and Simplified Acute Physiology score (SAPS II) were associated with lower RBP4 levels (r = -0.692, p = 0.024 and r = -0.670, p = 0.015, respectively). RBP4 was positively correlated with creatinine and body mass index, and negatively correlated with C-reactive protein and Global Initiative for Chronic Obstructive Lung Disease stage. Multivariate logistic regression showed that RBP4 was an independent mortality predictor of AECOPD (odds ratio: 0.926, p = 0.007). Analysis of the area under the receiver operating characteristic (AUC) curve showed that RBP4 showed good discrimination (AUC: 0.88; 95% confidence interval: 0.78-0.94; p = 0.008) in predicting mortality. RBP4 improved the prognostic accuracy of mortality for the APACHE II and SAPS II scores. CONCLUSIONS: Serum RBP4 levels are significantly reduced in elderly AECOPD patients. RBP4 might be a good predictive biomarker for mortality in elderly AECOPD patients in the intensive care unit.