Enhancing Fractionated Cancer Therapy: A Triple-Anthracene Photosensitizer Unleashes Long-Persistent Photodynamic and Luminous Efficacy.

Conventional photodynamic therapy (PDT) is often limited in treating solid tumors due to hypoxic conditions that impede the generation of reactive oxygen species (ROS), which are critical for therapeutic efficacy. To address this issue, a fractionated PDT protocol has been suggested, wherein light irradiation is administered in stages separated by dark intervals to permit oxygen recovery during these breaks. However, the current photosensitizers used in fractionated PDT are incapable of sustaining ROS production during the dark intervals, leading to suboptimal therapeutic outcomes (Table S1). To circumvent this drawback, we have synthesized a novel photosensitizer based on a triple-anthracene derivative that is designed for prolonged ROS generation, even after the cessation of light exposure. Our study reveals a unique photodynamic action of these derivatives, facilitating the direct and effective disruption of biomolecules and significantly improving the efficacy of fractionated PDT (Table S2). Moreover, the existing photosensitizers lack imaging capabilities for monitoring, which constraints the fine-tuning of irradiation parameters (Table S1). Our triple-anthracene derivative also serves as an afterglow imaging agent, emitting sustained luminescence postirradiation. This imaging function allows for the precise optimization of intervals between PDT sessions and aids in determining the timing for subsequent irradiation, thus enabling meticulous control over therapy parameters. Utilizing our novel triple-anthracene photosensitizer, we have formulated a fractionated PDT regimen that effectively eliminates orthotopic pancreatic tumors. This investigation highlights the promise of employing long-persistent photodynamic activity in advanced fractionated PDT approaches to overcome the current limitations of PDT in solid tumor treatment.

Elevated D-Dimer levels correlate with the development of hepatorenal syndrome and a poor outcome in patients with cirrhosis.

OBJECTIVES: Whether hemostatic status was correlated with the diverse types of acute kidney injury in cirrhotic patients is unclear. The present study aimed to investigate the relationship between hemostatic markers and the diverse types of acute kidney injury (AKI) in liver cirrhosis. PATIENTS AND METHODS: Cirrhotic patients with consecutive treatment at the First Affiliated Hospital of Medicine School, Zhejiang University, were pooled in a cohort. Their demographic and clinical data, biochemistry parameters and hemostatic markers were assessed to identify risk factors for the development and prognosis of AKI. RESULTS: A total of 773 cirrhotic patients were included in this cohort. Patients with hepatorenal syndrome (HRS) had significantly higher D-Dimer than those with the other types of AKI. In univariate COX regression, APTT, TT, INR, D-Dimer and Fib were correlated with the development of AKI, HRS and acute tubular necrosis (ATN), however, only D-Dimer remained independently associated with the development of AKI and HRS in multivariate COX regression. The area under the ROC curve of D-Dimer was 0.755 (95%CI, 0.718-0.793) in predicting the development of AKI, 0.879 (95%CI, 0.791-0.967) in predicting the development of HRS, respectively. D-Dimer was used for diagnosis of HRS with a sensitivity of 87.3% and specificity of 72.9% at the cutoff of 3.7 (mg/L FEU). Survival rates differed significantly between groups by D-Dimer level. CONCLUSIONS: Hemostatic markers were significantly associated with the diverse types of AKI. D-Dimer was an independent risk factor for HRS and correlated with a poor outcome in cirrhotic patients.

The Love-Hate Relationship Between TGF-ß Signaling and the Immune System During Development and Tumorigenesis.

Since TGF-ß was recognized as an essential secreted cytokine in embryogenesis and adult tissue homeostasis a decade ago, our knowledge of the role of TGF-ß in mammalian development and disease, particularly cancer, has constantly been updated. Mounting evidence has confirmed that TGF-ß is the principal regulator of the immune system, as deprivation of TGF-ß signaling completely abrogates adaptive immunity. However, enhancing TGF-ß signaling constrains the immune response through multiple mechanisms, including boosting Treg cell differentiation and inducing CD8+ T-cell apoptosis in the disease context. The love-hate relationship between TGF-ß signaling and the immune system makes it challenging to develop effective monotherapies targeting TGF-ß, especially for cancer treatment. Nonetheless, recent work on combination therapies of TGF-ß inhibition and immunotherapy have provide insights into the development of TGF-ß-targeted therapies, with favorable outcomes in patients with advanced cancer. Hence, we summarize the entanglement between TGF-ß and the immune system in the developmental and tumor contexts and recent progress on hijacking crucial TGF-ß signaling pathways as an emerging area of cancer therapy.

Serum free light chain is associated with histological activity and cirrhosis in patients with chronic hepatitis B.

BACKGROUND: The antiviral immune response is the main cause of hepatocyte damage and inflammatory necrosis. The serum free light chain, reflecting the immune function of B-cells, is strongly associated with inflammation and disease activity. We aimed to investigate the association of serum free light chain with the progression of chronic hepatitis B. METHODS: A total of 208 eligible chronic hepatitis B patients who had undergone a liver biopsy were studied. Serum free light chains of all patients were measured by turbidimetry using an immunoassay. Liver histology was assessed according to the METAVIR scoring system (which grades the stage of fibrosis on a five-point scale, F0 = no fibrosis to F4 = cirrhosis, and histological activity on a four-point scale, A0 = no activity to A3 = severe activity). The association of serum free light chains with histological activity and fibrosis progression was evaluated. RESULTS: The concentration of serum free light chains in CHB patients increased gradually with histological activity and fibrosis progression. The intensity of histological activity was significantly correlated with the serum free kappa chain (r = 0.658, P < 0.001) and the serum free lambda chain (0.675, P < 0.001). The stages of fibrosis were correlated with the serum free kappa chain (r = 0.683, P < 0.001) and serum free lambda chain (0.664, P < 0.001). After adjusting for age, sex and other synergic factors, the serum free kappa chain remained a potential risk factor, but the serum free lambda chain was no longer associated with liver cirrhosis. Similar to FIB-4 and RPR, the serum free kappa chain exhibited excellent performance in the prediction of liver cirrhosis. The AUCs of serum free Kappa chain, FIB-4 and RPR were 0.873, 0.880 and 0.895, respectively, which were significantly higher than those of the AAR and APRI (0.718 and 0.746). CONCLUSION: Our work revealed that serum free light chains were associated with histological activity and cirrhosis in chronic hepatitis B, which could play a crucial role in the immunopathogenesis of HBV-associated cirrhosis. In addition, free kappa light chain could be a useful predictor of liver cirrhosis.

Using Cell Type-Specific Genes to Identify Cell-Type Transitions Between Different in vitro Culture Conditions.

In vitro differentiation or expansion of stem and progenitor cells under chemical stimulation or genetic manipulation is used for understanding the molecular mechanisms of cell differentiation and self-renewal. However, concerns around the cell identity of in vitro-cultured cells exist. Bioinformatics methods, which rely heavily on signatures of cell types, have been developed to estimate cell types in bulk samples. The Tabula Muris Senis project provides an important basis for the comprehensive identification of signatures for different cell types. Here, we identified 46 cell type-specific (CTS) gene clusters for 83 mouse cell types. We conducted Gene Ontology term enrichment analysis on the gene clusters and revealed the specific functions of the relevant cell types. Next, we proposed a simple method, named CTSFinder, to identify different cell types between bulk RNA-Seq samples using the 46 CTS gene clusters. We applied CTSFinder on bulk RNA-Seq data from 17 organs and from developing mouse liver over different stages. We successfully identified the specific cell types between organs and captured the dynamics of different cell types during liver development. We applied CTSFinder with bulk RNA-Seq data from a growth factor-induced neural progenitor cell culture system and identified the dynamics of brain immune cells and nonimmune cells during the long-time cell culture. We also applied CTSFinder with bulk RNA-Seq data from reprogramming induced pluripotent stem cells and identified the stage when those cells were massively induced. Finally, we applied CTSFinder with bulk RNA-Seq data from in vivo and in vitro developing mouse retina and captured the dynamics of different cell types in the two development systems. The CTS gene clusters and CTSFinder method could thus serve as promising toolkits for assessing the cell identity of in vitro culture systems.

Evaluation of Sysmex XN-1000 hematology analyzer for cell count and screening of malignant cells of serous cavity effusion.

Over the years, with the advancement in hematology analyzer technology, the use of fluid analysis method has seen a drastic increase in clinical examinations. Cell counting and classification in independent body fluid analysis method are conducted by semiconductor laser flow cytometry and nucleic acid fluorescence staining techniques. This study is to evaluate the efficacy of Sysmex XN-1000 hematology analyzer in cell counting and to screen malignant cells with serous cavity effusion. Specimens (N = 206) with serous cavity effusion from our hospital were included in this study. Manual and instrumental methods for cell counting, nucleated cell classification, and high-fluorescent cells (HFC) were used in this study. The correlation between RBC, nucleated cell count (NUC), the percentages of polymorphonuclear cell (PMN%), and mononuclear cells (MN%) was statistically analyzed using manual and instrumental methods. The regression equations of RBC, NUC, PMN%, and MN% in the manual and instrumental methods were RBC y = 0.88x + 426.4; NUC y = 0.85x + 33.4; PMN% y = 0.91x + 4.2; and MN% y = 0.91x + 5.1. Correlation coefficient R was 0.99, 0.98, 0.90, and 0.90 (P < .001). ROC curve analysis showed that when the cut-off value of HFC% was 4.4% and HFC# was 24.5/µL, area under curve (AUC), sensitivity, specificity, and 95% confidence interval were 0.707, 0.792, 0.558, 0.637-0.777; 0.708, 0.753, 0.550, 0.635-0.780, respectively. XN-1000 hematology analyzer body fluid method can accurately and rapidly count cell and nucleated cell classification with serous cavity effusion. HFC can indicate the possible existence of malignant cells; however, further investigations are required to validate its efficacy.

Chronic myeloid leukemia patient with co-occurrence of BCR-ABL junction and JAK2 V617F mutation.

The JAK2 V617F mutation is common in patients with Philadelphia-negative chronic myeloproliferative neoplasms, but few cases of the JAK2 V617F mutation have been described in Philadelphia-positive chronic myeloid leukemia (CML) patients. Here, we report a 21-year-old female who presented with phenotype of CML in whom BCR-ABL transcript and JAK2V617F mutation co-occurred. These findings were determined through cytogenetic analysis, fluorescence in situ hybridization, and allele-specific (AS) PCR. The patient's BCR-ABL transcript disappeared after 6 months of treatment with imatinib, while the JAK2V617F mutation remained positive. We discuss this case with reference to the current literature.

RDW to platelet ratio: a novel noninvasive index for predicting hepatic fibrosis and cirrhosis in chronic hepatitis B.

OBJECTIVE: To develop a simple predictive model for significant fibrosis and cirrhosis in chronic hepatitis B (CHB) using the routine hematological parameters of a complete blood count. METHODS: A total of 458 eligible CHB patients who had undergone a liver biopsy were randomly divided into two cohorts: an estimation group (n = 310) and a validation group (n = 148). Liver histology was assessed according to the Metavir scoring scheme. All common demographics, hematological parameters, HBeAg status, HBV DNA, and liver biochemistry were analyzed. RESULTS: Based on routinely available clinical parameters (age, sex, HBeAg status, HBV DNA, common hematological parameters of a complete blood cell count), a model for predicting significant fibrosis (Metavir score ≥2) in the estimation group was derived using platelets and red cell distribution width (RDW), and another model for predicting cirrhosis (Metavir score = 4) was derived using platelets, RDW and hemoglobin. A novel index, the RDW to platelet ratio (RPR), was developed to amplify the opposing effects of liver fibrosis on the RDW and platelets. The AUCs of the RPR for predicting significant fibrosis and cirrhosis were 0.825 and 0.884, respectively, which is superior to the AAR, FIB-4 and APRI in the estimation group. Compared with the two derived models, the RPR has a comparable predictive power for significant fibrosis and cirrhosis. Using optimized cutoffs (0.10 and 0.16), the RPR accurately predicted 63.1% of cases with significant fibrosis and 73.7% of cases with cirrhosis and accurately excluded 85.5% of the cases with mild fibrosis and 93.0% of the cases with no cirrhosis. CONCLUSION: The RPR, a routinely available, inexpensive and easily calculated index, can predict significant fibrosis and cirrhosis in CHB patients with relatively high accuracy. The application of this index may reduce the need for liver biopsy in CHB patients.

Performance evaluation and result comparison of the automated hematology analyzers Abbott CD 3700, Sysmex XE 2100 and Coulter LH 750 for cell counts in serous fluids.

BACKGROUND: Automated hematology analyzers are used to perform cell counts in body fluids. However, little is known about how the results compare between different analyzers. METHODS: A single batch of serous fluid samples was used to evaluate the cell counting performance of 3 hematology analyzers: CD 3700, XE 2100, and LH 750. Two hundred and seventy four serous fluid samples were used to evaluate the accuracy of the analyzers and to compare the results between different analyzers. RESULTS: The precision and linearity of the 3 analyzers were acceptable for both white blood cell counts and red blood cell counts with a low carryover rate. The limits of detection for white blood cells with the CD 3700, XE 2100, and LH 750 analyzers were 0.033×10(9)/l, 0.07×10(9)/l, and 0.20×10(9)/l, respectively. Performing background counts had no influence on cell counts for any of the analyzers. For those samples over the limit of detection, there was agreement between the automated and manual counting methods. There were also reasonably comparable cell count results between the 3 analyzers. CONCLUSIONS: When the serous fluid cell counts are over the limit of detection, the analyzers produce accurate test results. Additionally, the results are comparable between the 3 hematology analyzers.

Effect of telbivudine therapy on the cellular immune response in chronic hepatitis B.

Weak T-cell reactivity to the hepatitis B virus (HBV) is believed to be the dominant cause of chronic HBV infection. Several lines of experimental evidence suggest that treatment with telbivudine increases the rate of HBV e antigen (HBeAg) loss, undetectable HBV DNA, and normalization of serum alanine aminotransferase (ALT) in chronic hepatitis B patients (CHB). However, it is still unclear how early antiviral therapy affects cellular immune responses during sustained telbivudine treatment. In order to investigate this issue, we measured detailed prospective clinical, virological, and biochemical parameters, and we examined the frequency of T cell subgroups as well as the ability of peripheral blood mononuclear cells (PBMC) to respond to stimuli at five protocol time points for 51 CHB patients who received telbivudine therapy for one year. The preliminary data from this study revealed that effective-treated patients showed an increased frequency of peripheral blood CD4(+)T lymphocytes, an augmented proliferative response of HBV-specific T-cells to the hepatitis B core antigen (HBcAg), and the induction of cytokines, such as interferon gamma (IFN-γ), tumour necrosis factor alpha (TNF-α) release at the site of infection compared to non-responsive patients. Enhanced HBV-specific T-cell reactivity to telbivudine therapy, which peaked at treatment week 12, was confined to a subgroup of effective-treated patients who achieved greater viral suppression.

Simultaneous detection of influenza A, influenza B, and respiratory syncytial viruses and subtyping of influenza A H3N2 virus and H1N1 (2009) virus by multiplex real-time PCR.

A multiplex real-time PCR assay was developed to simultaneously detect and discriminate influenza A virus subtypes, including novel H1N1 (2009) and seasonal H3N2 virus, influenza B virus, and respiratory syncytial virus (RSV) in a single test tube, with detection sensitivity and specificity of 99% and 100%, respectively, for the four pathogens.

[Prokaryotic expression of HPV 16L1 and optimization of expression conditions].

OBJECTIVE: To construct the HPV16 L1 prokaryotic expression plasmid and to optimize its expression. METHODS: A pair of primers was designed according to plasmid sequences of pGEX-KG and the HPV16L1 genes published by GeneBank. The DNA fragment of 1500 bp was amplified by PCR from the HPV recombinant plasmid with HPV16L1 gene, then cloned into pGEX-KG and transformed into the host E.coli strain JM109. The pGEX-KG-HPV16L1 plasmid was taken and transformed into BL21(DE3) for expression. Induced by IPTG at 37 degree, the expression product of HPV16L1 gene was identified by SDS-PAGE and Western blot. RESULTS: HPV16L1 fusion protein was expressed successfully in the form of inclusion bodies. The molecular weight was 83 kD. Meanwhile, the optimum condition of HPV16L1 fusion protein expression was induced with 1.0 mmol*L(-1) IPTG for 4 h. The fusion protein reacted specifically with antibodies against HPV16L1. CONCLUSION: The prokaryotic expression vector of HPV16L1 gene has been constructed and expressed in E.coli successfully.

The prevalence of microalbuminuria and its relationships with the components of metabolic syndrome in the general population of China.

BACKGROUND: Few studies have examined the relationships between the prevalence of microalbuminuria and the metabolic risk factors in the general population of China. We performed a population based study to investigate the prevalence of microalbuminuria and its relationships with the components of the metabolic syndrome in Hangzhou, China. METHODS: The subjects of this cross-sectional study were the individuals from 19 to 87 y. The metabolic syndrome was defined based on the criteria of the Chinese Diabetes Society (CDS). Microalbuminuria was defined as a urine albumin-creatinine ratio of 30 to 300 mg/g. RESULTS: A total of 2985 subjects (average age of 44 y) were analyzed. Among them, the prevalence of the metabolic syndrome and microalbuminuria was 12.6% and 8.8%, respectively. Microalbuminuria prevalence rate was significantly higher in the population >60 y than <60 y. The prevalence of MAU in the group with metabolic abnormalities was significantly higher than the control group, and the prevalence rate of MAU in the metabolic syndrome group reached up to 20.3%. There was a significantly positive correlation between the prevalence of microalbuminuria and the corresponding components of the metabolic syndrome (P<0.001). CONCLUSIONS: Microalbuminuria was highly prevalent in the middle-aged and elderly Chinese population in the city of Hangzhou. There is an increasing likelihood of having microalbuminuria if subjects have the metabolic syndrome. Early screening strategies for prevention and treatment of MAU are strongly suggested, especially in the population >60 y and the ones with metabolic abnormalities.

[Numerical simulations of spatial and temporal characteristics of airborne dust over Asia during springs of 2000 to 2002].

Spatial and temporal characteristics of airborne dust over Asia during springs of 2000, 2001 and 2002 were simulated with a mineral dust entrainment and deposition model (DEAD) embedded in a global model of atmospheric transport and chemistry (MATCH) using the real-time meteorological data as forcing fields. The results show a good agreement of the pattern of model-simulated atmospheric dust concentration with the distribution of surface-observed spring dust storm frequency and a significant correlation between the simulated dust aerosol optical depth (DAOD) and satellite-observed aerosol index (AI). These results validate applicability of the integrated model in simulating dust entrainment, transportation and deposition and describing spatial and temporal characteristics of dust loading over the Asian continent. In addition, an attempt was made to explore possible paths of dust transportation by use of correlation analyses between the simulated dust emission flux (DEF) and DAOD.

The effect of transformation on the virulence of Streptococcus pneumoniae.

Although pneumococcus is one of the most frequently encountered opportunistic pathogen in the world, the mechanisms responsible for its infectiveness have not yet been fully understood. In this paper, we have attempted to characterize the effects of pneumococcal transformation on the pathogenesis of the organism. We constructed three transformation-deficient pneumococcal strains, which were designated as Nos. 1d, 2d, and 22d. The construction of these altered strains was achieved via the insertion of the inactivated gene, comE, to strains 1, 2 and 22. We then conducted a comparison between the virulence of the transformation-deficient strains and that of the wild-type strains, via an evaluation of the ability of each strain to adhere to endothelial cells, and also assessed psaA mRNA expression, and the survival of hosts after bacterial challenge. Compared to what was observed with the wild-type strains, our results indicated that the ability of all of the transformation-deficient strains to adhere to the ECV304 cells had been significantly reduced (p < 0.05), the expression of psaA mRNA was reduced significantly (p < 0.05) in strains 2d and 22d, and the median survival time of mice infected with strains 1d and 2d was increased significantly after intraperitoneal bacterial challenge (p < 0.05). The results of our study also clearly indicated that transformation exerts significant effects on the virulence characteristics of S. pneumoniae, although the degree to which this effect is noted appears to depend primarily on the genetic background of the bacteria.