Association between decreased grip strength in preschool children and the COVID-19 pandemic: an observational study from 2015 to 2021.

BACKGROUND: Coronavirus disease 2019 (COVID-19) has reduced people's physical activity. It is essential to accumulate knowledge regarding the influence of COVID-19 on the stimulation of physical fitness and physical functions. Several studies have reported the effects of COVID-19 on physical fitness; however, there are very few reports regarding preschoolers. This study aimed to compare the physical fitness of preschoolers before and during the COVID-19 pandemic to clarify the effects of curtailment of outings implemented to control the pandemic on physical fitness among preschoolers. METHODS: The subjects were 593 Japanese preschool children enrolled at a kindergarten during 2015-2019 and in 2021 who received a physical fitness test. Children enrolled in 2020 who did not receive a physical fitness test because of the COVID-19 pandemic were excluded. The physical fitness test included grip strength, standing long jump, and a 25-m run. The relationship between physical fitness level and survey year was analyzed using a general linear model, with grip strength and standing long jump as dependent variables, year of study as the independent variable, and sex and age in months as adjusted variables. Kruskal-Wallis test was used to analyze data for the 25-m run. Multiple comparisons were used to compare fitness levels between 2021 (during the COVID-19 pandemic) with levels in previous years. RESULTS: Significant relationships were found between survey year and each of grip strength (p < 0.001), standing long jump (p < 0.05), and 25-m run (p < 0.001) among the overall subjects. Grip strength was significantly lower in 2021 compared with the 2016-2019 period. Similarly, sub-stratification analysis by sex showed that grip strength was lower in 2021 than in previous survey years, in both sexes. However, there was no difference in standing long jump or 25-m run times between before and during the pandemic among the overall subjects or according to sex. CONCLUSIONS: These findings indicate that the COVID-19 pandemic has had a negative effect on the development of muscle strength in preschoolers, and suggest the need to develop strategies that could promote the development of muscle strength of preschool children when limitations are placed on activity during prolonged infectious disease pandemics.

RNA Modification in Inflammatory Bowel Diseases.

Inflammatory bowel disease (IBD) is a chronic inflammatory disorder characterized by damage to the intestinal mucosa, which is caused by a combination of factors. These include genetic and epigenetic alterations, environmental influence, microorganism interactions, and immune conditions. Some populations with IBD show a cancer-prone phenotype. Recent studies have provided insight into the involvement of RNA modifications in the specific pathogenesis of IBD through regulation of RNA biology in epithelial and immune cells. Studies of several RNA modification-targeting reagents have shown preferable outcomes in patients with colitis. Here, we note a new awareness of RNA modification in the targeting of IBD and related diseases, which will contribute to early diagnosis, disease monitoring, and possible control by innovative therapeutic approaches.

Establishment of a reference single-cell RNA sequencing dataset for human pancreatic adenocarcinoma.

Single-cell RNA sequencing (scRNAseq) has been used to assess the intra-tumor heterogeneity and microenvironment of pancreatic ductal adenocarcinoma (PDAC). However, previous knowledge is not fully universalized. Here, we built a single cell atlas of PDAC from six datasets containing over 70 samples and >130,000 cells, and demonstrated its application to the reanalysis of the previous bulk transcriptomic cohorts and inferring cell-cell communications. The cell decomposition of bulk transcriptomics using scRNAseq data showed the cellular heterogeneity of PDAC; moreover, high levels of tumor cells and fibroblasts were indicative of poor-prognosis. Refined tumor subtypes signature indicated the tumor cell dynamics in intra-tumor and their specific regulatory network. We further identified functionally distinct tumor clusters that had close interaction with fibroblast subtypes via different signaling pathways dependent on subtypes. Our analysis provided a reference dataset for PDAC and showed its utility in research on the microenvironment of intra-tumor heterogeneity.

EpisomiR, a New Family of miRNAs, and Its Possible Roles in Human Diseases.

MicroRNAs (miRNAs) are synthesized through a canonical pathway and play a role in human diseases, such as cancers and cardiovascular, neurodegenerative, psychiatric, and chronic inflammatory diseases. The development of sequencing technologies has enabled the identification of variations in noncoding miRNAs. These miRNA variants, called isomiRs, are generated through a non-canonical pathway, by several enzymes that alter the length and sequence of miRNAs. The isomiR family is, now, expanding further to include episomiRs, which are miRNAs with different modifications. Since recent findings have shown that isomiRs reflect the cell-specific biological function of miRNAs, knowledge about episomiRs and isomiRs can, possibly, contribute to the optimization of diagnosis and therapeutic technology for precision medicine.

Single-cell RNA sequencing unravels heterogeneity of skeletal progenitors and cell-cell interactions underlying the bone repair process.

Introduction: Activation of skeletal progenitors upon tissue injury and the subsequent cell fate specification are tightly coordinated in the bone repair process. Although known osteoimmunological signaling networks play important roles in the microenvironment of the bone defect sites, the molecular mechanism underlying the bone repair process has not been fully understood. Methods: To better understand the behavior of the skeletal progenitors and the heterogeneity of the cells during bone repair at the microenvironmental level, we performed a combinatorial analysis consisting of lineage tracing for skeletal progenitors using the Sox9-CreERT2;R26R tdTomato mouse line followed by single-cell RNA sequencing (scRNA-seq) analysis using a mouse model of calvarial bone repair. To identify a therapeutic target for bone regeneration, further computational analysis was performed focusing on the identification of the cell-cell interactions, followed by pharmacological assessments with a critical-size calvarial bone defect mouse model. Results: Lineage tracing analysis showed that skeletal progenitors marked by Sox9 were activated upon bone injury and contributed to bone repair by differentiating into osteoblasts. The scRNA-seq analysis characterized heterogeneous cell populations at the bone defect sites; the computational analysis predicted a bifurcated lineage from skeletal progenitors toward osteogenic and adipogenic lineages. Chemokine C-C motif ligand 9 (Ccl9) was identified as a signaling molecule that regulates bone regeneration in the mouse model, possibly through the regulation of adipogenic differentiation at the bone defect site. Conclusion: Multipotential skeletal progenitors and the direction of the cell differentiation were characterized at single cell resolution in a mouse bone repair model. The Ccl9 signaling pathway may be a key factor directing osteogenesis from the progenitors in the model and may be a therapeutic target for bone regeneration.

[A case of combined hepatocellular and cholangiocarcinoma showing tumor growth and invasion to the diaphragm during interferon-alpha and 5-fluorouracil combined intra-arterial chemotherapy].

We report a case of combined hepatocellular and cholangiocarcinoma showing tumor growth and invasion to the diaphragm during interferon-alpha (IFN-alpha) and 5-fluorouracil (5-FU) combined intra-arterial chemotherapy. A 63-year-old female was in hospital for treatment of hepatic tumor in left lobe with portal venous tumor thrombus. She was treated by IFN-alpha/5-FU chemotherapy, and the tumor was significantly reduced. For 28 months, the tumor was successfully treated by IFN-alpha/5-FU chemotherapy. But, thereafter the abdominal computed tomography showed the tumor re-growth and invasion to the diaphragm. Then the patient underwent the tumor resection. The histological findings were consistent with combined hepatocellular and cholangiocarcinoma. Immunohistological examination revealed a heterogeneous expression of IFN-alpha receptor 2. This case suggested that the growth of cancer cells without sensitivity to IFN-alpha/5-FU chemotherapy and the blood supply via the diaphragm led the relapse of IFN-alpha/5-FU chemotherapy.

First phase viral kinetic parameters and prediction of response to interferon alpha-2b/ribavirin combination therapy in patients with chronic hepatitis C.

The aim of the present study was to assess parameters in early phase HCV dynamics for predicting the outcome of interferon (IFN)/ribavirin combination therapy in patients with chronic hepatitis C (CH-C). Sixty-five CH-C patients who received IFN alpha-2b/ribavirin combination therapy were enrolled. The serum levels of HCV RNA 0h and 3 months after commencing therapy were serially quantified. HCV kinetic parameters such as quantity, ratio of decline, and half-life were analyzed. In genotype 1 patients, both the quantity and the ratio of decline of HCV RNA 24h after the start of therapy were useful predictors of a poor response. No patients who had serum HCV RNA above 200KIU/ml 24h after the start of therapy achieved a sustained viral response (SVR). In genotype 2 patients, conversely, these two parameters were predictors of a sustained viral response. The efficacy of these parameters in predicting the outcome of therapy was comparable to that of the disappearance of HCV RNA from sera at 4 weeks. These results demonstrate that parameters of HCV kinetics 24h after the start of therapy are useful for the early prediction of outcome in response to IFN alpha-2b/ribavirin combination therapy.

Prediction of breakthrough hepatitis due to lamivudine-resistant hepatitis B virus by a sensitive semiquantitative assay using peptide nucleic acids.

OBJECTIVE: The aim of this study was to predict breakthrough hepatitis and analyze the dynamics of lamivudine-resistant hepatitis B virus in patients treated with lamivudine. METHODS: Fifty-five chronic hepatitis B patients treated with lamivudine were included. The emergence of YMDD motif mutants was detected by peptide nucleic acid (PNA) mediated PCR clamping with a detection limit of 10(1) YMDD mutants. We then performed a semiquantitative PCR assay of subjects in whom YMDD mutants were detected. This assay detects 10(2.7)-10(7.7) copies of mutant virus per 1 ml of serum. RESULTS: YMDD mutants were detected in 28 (51%) of the 55 patients. Eight patients stopped medication before viral breakthrough. YMDD mutants appeared transiently despite the continuance of lamivudine therapy in 12 patients. In all 8 patients with breakthrough hepatitis, the quantities of YMDD mutants ranged from 10(2.7)-10(4.7) copies/ml in the two to three months before clinical breakthrough. In contrast, in 12 patients without viral breakthrough, there were always less than 10(2.7) copies/ml YMDD mutants. CONCLUSIONS: Lamivudine-resistant viruses sometimes disappear even during lamivudine administration. Our sensitive quantitative assay proved useful for early detection of YMDD mutants and a threshold of 10(2.7) copies/ml is suggested for predicting viral breakthrough.

A follow-up study to determine the value of liver biopsy and need for antiviral therapy for hepatitis C virus carriers with persistently normal serum aminotransferase.

BACKGROUND/AIMS: Long-term follow-up study was performed to identify the candidates for antiviral therapy for hepatitis C virus (HCV) infection among carriers with persistently normal aminotransferase (ALT< or = 30 U/L) levels (PNAL). METHODS: One hundred and twenty-nine HCV carriers with PNAL who underwent liver biopsy and had platelet count over 150,000/microl were entered and 69 were followed for over 5 years. Thirty-five patients underwent serial liver biopsies. Serum ferritin and thioredoxin levels were also determined. RESULTS: Seventeen patients had normal liver histology, 10 had moderate chronic hepatitis and the remainder 102 had mild hepatitis. Serum ferritin and thioredoxin levels were normal. The mean follow-up period for the 69 patients was 8.5 years. Of these 69 patients, 10 had persistently normal ALT levels (group A), 39 had transient elevation of ALT (group B), and 20 changed to symptomatic chronic hepatitis (group C). The rate of progression of fibrosis for groups A, B, and C were 0.05, 0.04, and 0.08, respectively. Hepatocellular carcinoma was not diagnosed in any of the patients. CONCLUSIONS: Around 90% of HCV carriers with PNAL have normal to mild liver histology. This long-term follow-up study demonstrated that 30% of such carriers became candidates for antiviral therapy within 5 years.

Cell growth after withdrawal of gefitinib ("Iressa", ZD1839) in human lung cancer cells.

Gefitinib ("Iressa", ZD1839) is a selective epidermal growth factor receptor-tyrosine kinase inhibitor, which blocks signal transduction pathways implicated in proliferation and survival of cancer cells. However, in vitro and in vivo studies concerning cell growth after withdrawal of gefitinib are limited. To determine whether cancer cells would resume proliferation upon removal of gefitinib, an in vitro study was undertaken using 4 human non-small cell lung cancer cell lines. With immunocytochemical staining and Western blot analysis, we confirmed positive expression of EGFR in these cell lines. We next evaluated growth inhibition before and after withdrawal of gefitinib. After incubation with 0-100 microM gefitinib for 24 h, medium containing gefitinib was removed, followed by addition of fresh growth medium to cell cultures. MTT assays were performed daily over a 6-day time course. Even at very low levels of gefitinib (<1 microM), these 4 TKB cells had 1-10% growth inhibition, respectively. With these levels of gefitinib, continued inhibitory effects after withdrawal of getitinib were observed in 3 of the 4 cell lines. Furthermore, none of the 4 cell lines, including the cell line which had abolished growth inhibition, showed accelerated re-growth rate after the withdrawal of gefitinib even in these exposure conditions. Based on our in vitro experiments, additional in vivo studies, which can compare the pre- and post-treatment growth rate, will be necessary to help better understand the mechanisms behind cell growth recovery after temporary gefitinib treatment.

Circuit training for elderly patients with chronic obstructive pulmonary disease: a preliminary study.

To examine the efficacy of the circuit training in elderly patients with chronic obstructive pulmonary disease (COPD), we evaluated muscle forces of the upper and lower extremities and respiratory muscles, exercise tolerance and quality of life (QOL) before and after the circuit training in 10 male patients with mild to severe COPD. The circuit training improved muscle forces of the upper and lower extremities and abdominal muscles (P < 0.05), and 6 min walking distance (P < 0.05). Emotional function and mastery in the Chronic Respiratory Disease Questionnaire scores (P < 0.05) were also improved after the circuit training. The circuit training designed in the present study was effective to improve the QOL in elderly COPD patients.

Cytokeratin 19 fragment in patients with nonmalignant respiratory diseases.

STUDY OBJECTIVE: Cytokeratin 19 fragment (CYFRA) is a specific tumor marker in patients with lung cancer; however, it has been reported that serum CYFRA levels are elevated in some patients with nonmalignant respiratory diseases such as interstitial pulmonary fibrosis (IPF) and collagen disease-associated pulmonary fibrosis (CDPF). To investigate the serum CYFRA levels in nonmalignant respiratory diseases in detail, we studied 413 patients with respiratory diseases. DESIGN: Retrospective study. SETTING: University hospital. PATIENTS: Four hundred thirteen patients with nonmalignant respiratory diseases and lung cancer. MEASUREMENTS AND RESULTS: Serum CYFRA levels were measured with a commercially available enzyme immunoassay kit. Immunohistochemical study was performed using monoclonal antibody Ks 19.1 (Rosch Diagnosica; Bern, Switzerland) on surgically resected or autopsied lung specimens. Gel electrophoresis and immunoblotting was performed with serum samples. In 149 patients with nonmalignant diseases except IPF and CDPF, the ratio of patients with > 3.5 ng/mL of serum CYFRA was 13.4%. In 13 of 30 patients (43.3%) with IPF and CDPF, the serum CYFRA levels were abnormally elevated. The 95th percentile serum CYFRA level of the patients with nonmalignant respiratory diseases was 6.2 ng/mL, and none of them had CYFRA levels > 20.3 ng/mL. Survival in patients with IPF and CDPF with elevated serum CYFRA levels were significantly lower than in those with normal range (p = 0.0335). Western blotting using serum from nonmalignant lung diseases and patients with lung cancer showed no apparent difference between them. An immunohistochemical study indicated CYFRA was selectively expressed in the pulmonary epithelial cells that covered the remodeling alveolar septi in nonmalignant respiratory disease. CONCLUSION: Serum CYFRA was elevated in some nonmalignant respiratory diseases, especially in IPF and CDPF. The value of serum CYFRA would reflect the severity of lung injury in nonmalignant respiratory diseases and might be related to the prognosis in patients with IPF and CDPF.

Plasma orexin-A levels and body composition in COPD.

STUDY OBJECTIVE: To study the role of orexins in regulating body composition in patients with COPD. DESIGN: Prospective study. PATIENTS AND MEASUREMENTS: We measured the plasma concentration of orexin-A in 20 patients with COPD and compared the results to those obtained from 10 age-matched control subjects. Patients with COPD were classified into two groups based on their body mass index (BMI): a normal weight (NW) group (BMI > 20) and an underweight (UW) group (BMI < 20). RESULTS: The plasma orexin-A level was significantly lower in patients with COPD than in control subjects. In patients with COPD, the level was significantly lower in the UW group than in the NW group. Plasma orexin-A levels significantly correlated with BMI and fat mass values, but there was no significant relationship between plasma orexin-A levels and the fat-free mass of patients with COPD. CONCLUSION: These results suggest that orexin-A levels are altered with weight loss and changes in body composition in patients with COPD.

Effect of heavy-ion radiotherapy on pulmonary function in stage I non-small cell lung cancer patients.

STUDY OBJECTIVES: The purpose of this study was to investigate the effect of heavy-ion radiotherapy on pulmonary function in patients with clinical stage I non-small cell lung cancer (NSCLC) patients. DESIGN: Retrospective study. SETTING: Research Center Hospital for Charged Particle Therapy, National Institute of Radiological Sciences, Chiba, Japan. PATIENTS: From a total of 81 patients who were not candidates for surgical resection due to medical reasons or patient refusal, and who were treated with carbon beam radiotherapy from October 1994 to February 1999, the 52 patients who had completed the repeat overall pulmonary function tests at 6 and 12 months after undergoing heavy-ion radiotherapy were examined. The total heavy-ion irradiation dose ranged from 59.4 to 95.4 photon gray equivalents (GyE), with a mean dose of 76.2 GyE. INTERVENTIONS AND MEASUREMENT: Pulmonary function was evaluated prior to heavy-ion radiotherapy and at 6 and 12 months after heavy-ion radiotherapy. Comparisons of all pulmonary function indexes between, before, and at 6 and 12 months after heavy-ion radiotherapy were made using repeated-measures analysis of variance using the Dunnett test for post hoc comparison. RESULTS: A statistically significant decrease in FEV(1) and total lung capacity was detected at both 6 and 12 months after the patient had undergone heavy-ion radiotherapy. No significant decreases in other pulmonary function indexes in patients were observed at either 6 or 12 months after heavy-ion radiotherapy. The magnitude of the decrease in all pulmonary function indexes was < 8% at both 6 and 12 months after heavy-ion radiotherapy. CONCLUSIONS: These findings suggest that heavy-ion radiotherapy is feasible for stage I NSCLC patients without a severe loss of pulmonary function.

Age-related changes in plasma orexin-A concentrations.

To study the effect of aging on orexin-A, we measured plasma orexin-A concentrations in 82 healthy volunteers (55 men and 27 women) over a wide range of ages (mean 50.2 +/- 13.9 years, ranging from 23 to 79 years). Correlation analyses revealed that plasma orexin-A concentrations correlated with age (r = 0.50, P < 0.01). When comparing three age groups, it appeared that the concentrations in the group of more than 60 years were significantly higher than those in the group of less than 39 years in both genders (P < 0.05). These findings suggest that orexin-A could be involved in aging in a healthy population.