K33 only mutant ubiquitin augments bone marrow-derived dendritic cell-mediated CTL priming via PI3K-Akt pathway.

To explore the effect of K33 only mutant ubiquitin (K33O) on bone marrow-derived dendritic cells' (BMDCs') maturity, antigen uptake capability, surface molecule expressions and BMDC-mediated CTL priming, and further investigate the role of PI3K-Akt engaged in K33O-increased BMDC maturation, antigen uptake and presentation, surface molecule expressions and BMDC-based CTL priming. BMDCs were conferred K33O and other ubiquitin mutants (K33R, K48R, K63R-mutant ubiquitin) incubation or LY294002 and wortmannin pretreatment. PI3K-Akt phosphorylation, antigen uptake, antigenic presentation and CD86/MHC class I expression in BMDC were determined by western blot or flow cytometry. BMDC-based CTL proliferation and priming were determined by in vitro mixed lymphocyte reaction (MLR), ex vivo enzyme-linked immunospot assay (Elispot) and flow cytometry with intracellular staining, respectively. The treatment with K33O effectively augmented PI3K-Akt phosphorylation, BMDCs' antigen uptake, antigenic presentation, CD86/MHC class I and CD11c expressions. MLR, Elispot and flow cytometry revealed that K33O treatment obviously enhanced CTL proliferation, CTL priming and perforin/granzyme B expression. The pretreatment with PI3K-Akt inhibitors efficiently abrogated K33O's effects on BMDC. The replenishment of K33 only mutant ubiquitin augments BMDC-mediated CTL priming in bone marrow-derived dendritic cells via PI3K-Akt signalling.

K48- and K27-mutant ubiquitin regulates adaptive immune response by affecting cross-presentation in bone marrow precursor cells.

K48-linked ubiquitination determines antigen degradation and plays vital roles in the process of cross-presentation of bone marrow precursor cell (BMPC)-derived dendritic cells (DCs). Although previous studies revealed that K48 and K27-linked ubiquitination regulates innate immunity, the exact roles of K48 and K27-linked ubiquitination in cross-presentation and BMPC-based adaptive immunity are still uncertain. In this study, we investigated the effects of K48- and K27-mutant ubiquitin (Ub) on BMPC-based adaptive immune response by observing the effects of MG132, Ub deficiency, and K48/K27-mutant Ub on cross-presentation, T cell proliferation, IFN-γ secretion, BMPC-based CTL priming, and thereby the efficiency of cytolytic capacity of BMPC-activate T cells. We demonstrated that MG132, Ub deficiency, and K48-mutant Ub impair cross-presentation, T cell proliferation, IFN-γ secretion, BMPC-based CTL priming, and the cytolytic capacity of BMPC-activated T cells. Moreover, although K27-only Ub decreases cross-presentation, the replenishment of K27-mutant Ub restores Ub deficiency impaireds the abilities of T cell proliferation, IFN-γ secretion, CTL priming, and the cytolytic capacity of BMPC-activated T cells. Thus, these data suggest that K48- and K27-linked ubiquitination contributes to BMPC-mediated adaptive immune response by affecting BMPC cross-presentation and the cytolytic capacity by up-regulating both perforin and granzyme B in BMPC-activated T cells. K48- and K27-mutant Ub might have potential clinical therapeutic function in adaptive immune response-associated diseases.

Hysteropreservation versus hysterectomy in uterine prolapse surgery: a systematic review and meta-analysis.

INTRODUCTION AND HYPOTHESIS: Hysteropreservation and hysterectomy for uterine prolapse have been compared in several randomized controlled trials (RCTs), as the best treatment has not been definitively determined. This study aimed to summarize the available evidence in RCTs of hysteropreservation versus hysterectomy. METHODS: We performed electronic searches in the PubMed, Embase, Cochrane Library, and China National Knowledge Infrastructure databases for eligible RCTs from inception to June 2020. The relative risks (RRs) and weighted mean differences (WMDs) with corresponding 95% confidence intervals (CIs) were calculated for categorical and continuous variables using random-effects models. RESULTS: Twelve RCTs involving 1177 patients were selected for meta-analysis. There were no significant differences between hysteropreservation and hysterectomy for the incidences of recurrence (RR, 0.55; 95% CI, 0.26-1.19; P = 0.130) and reoperation (RR, 1.15; 95% CI, 0.56-2.37; P = 0.705). Moreover, neither hysteropreservation nor hysterectomy had any significant effect on the risk of constipation (RR, 0.72; 95% CI, 0.15-3.46; P = 0.681), voiding dysfunction (RR, 0.99; 95% CI, 0.54-1.84; P = 0.981), intraoperative bleeding (RR, 0.35; 95% CI, 0.05-2.26; P = 0.271), upper leg dullness (RR, 0.70; 95% CI, 0.15-3.17; P = 0.643), dyspareunia (RR, 1.47; 95% CI, 0.69-3.13; P = 0.317), and wound infection (RR, 1.38; 95% CI, 0.24-7.80; P = 0.714). Furthermore, hysteropreservation was associated with less intraoperative blood loss (WMD, -25.68; 95% CI, -44.39 to -6.96; P = 0.007), shorter duration of surgery (WMD, -11.30; 95% CI, -19.04 to -3.55; P = 0.004), and shorter duration of hospitalization (WMD, -0.63; 95% CI, -1.10 to -0.16; P = 0.009) compared with hysterectomy. CONCLUSION: This study found that both hysteropreservation and hysterectomy have similar effects on recurrence and reoperation rates, while hysteropreservation was superior to hysterectomy in reducing intraoperative blood loss and shortening the duration of surgery and hospitalization.

K48-Linked Ubiquitination Contributes to Nicotine-Augmented Bone Marrow-Derived Dendritic-Cell-Mediated Adaptive Immunity.

K48-linked ubiquitination determining antigen degradation and the endosomal recruitments of p97 and Sec61 plays vital roles in dendritic cell (DC) cross-presentation. Our previous studies revealed that nicotine treatment increases bone marrow-derived dendritic cell (BM-DC) cross-presentation and promotes BM-DC-based cytotoxic T lymphocyte (CTL) priming. But the effect of nicotine on K48-linked ubiquitination and the mechanism of nicotine-increased BM-DC cross-presentation are still uncertain. In this study, we first demonstrated that ex vivo nicotine administration obviously increased K48-linked ubiquitination in BM-DC. Then, we found that K48-linked ubiquitination was essential for nicotine-augmented cross-presentation, BM-DC-based CTL priming, and thereby the superior cytolytic capacity of DC-activated CTL. Importantly, K48-linked ubiquitination was verified to be necessary for nicotine-augmented endosomal recruitments of p97 and Sec61. Importantly, mannose receptor (MR), which is an important antigenic receptor for cross-presentation, was exactly catalyzed with K48-linked ubiquitination by the treatment with nicotine. Thus, these data suggested that K48-linked ubiquitination contributes to the superior adaptive immunity of nicotine-administrated BM-DC. Regulating K48-linked ubiquitination might have therapeutic potential for DC-mediated immune therapy.

Human Ccr4 and Caf1 Deadenylases Regulate Proliferation and Tumorigenicity of Human Gastric Cancer Cells via Modulating Cell Cycle Progression.

Cancer cells generally have reprogrammed gene expression profiles to meet the requirements of survival, continuous division, and metastasis. An interesting question is whether the cancer cells will be affected by interfering their global RNA metabolism. In this research, we found that human Ccr4a/b (hCcr4a/b) and Caf1a/b (hCaf1a/b) deadenylases, the catalytic components of the Ccr4-Not complex, were dysregulated in several types of cancers including stomach adenocarcinoma. The impacts of the four deadenylases on cancer cell growth were studied by the establishment of four stable MKN28 cell lines with the knockdown of hCcr4a/b or hCaf1a/b or transient knockdown in several cell lines. Depletion of hCcr4a/b or hCaf1a/b significantly inhibited cell proliferation and tumorigenicity. Mechanistic studies indicated that the cells were arrested at the G2/M phase by knocking down hCaf1a, while arrested at the G0/G1 phase by depleting hCaf1b or hCcr4a/b. The four enzymes did not affect the levels of CDKs and cyclins but modulated the levels of CDK-cyclin inhibitors. We identified that hCcr4a/b, but not hCaf1a/b, targeted the p21 mRNA in the MKN28 cells. Furthermore, depletion of any one of the four deadenylases dramatically impaired processing-body formation in the MKN28 and HEK-293T cells. Our results highlight that perturbating global RNA metabolism may severely affect cancer cell proliferation, which provides a potential novel strategy for cancer treatment.

A novel F30S mutation in γS-crystallin causes autosomal dominant congenital nuclear cataract by increasing susceptibility to stresses.

Despite of increasingly accumulated genetic variations of autosomal dominant congenital cataracts (ADCC), the causative genes of many ADCC patients remains unknown. In this research, we identified a novel F30S mutation in γS-crystallin from a three-generation Chinese family with ADCC. The patients possessing the F30S mutation exhibited nuclear cataract phenotype. The potential molecular mechanism underlying ADCC by the F30S mutation was investigated by comparing the structural features, stability and aggregatory potency of the mutated protein with the wild type protein. Spectroscopic experiments indicated that the F30S mutation did not affect γS-crystallin secondary structure compositions, but modified the microenvironments around aromatic side-chains. Thermal and chemical denaturation studies indicated that the mutation destabilized the protein and increased its aggregatory potency. The mutation altered the two-state unfolding of γS-crystallin to a three-state unfolding with the accumulation of an unfolding intermediate. The almost identical values in the changes of Gibbs free energies for transitions from the native state to intermediate and from the intermediate to unfolded state suggested that the mutation probably disrupted the cooperativity between the two domains during unfolding. Our results expand the genetic variation map of ADCC and provide novel insights into the molecular mechanism underlying ADCC caused by mutations in ß/γ-crystallins.

Ex vivo IL-15 replenishment augments bone marrow precursor cell-mediated adaptive immunity via PI3K-Akt pathway.

This study tested the hypothesis that PI3K-Akt activity contributes to the superior immune function of IL-15-administrated bone marrow precursor cells (BMPC). Our previous studies revealed that PI3K-Akt play vital role in dendritic cells (DCs) cross-presentation and DC-based CTL priming. Despite the fact that IL-15 serves multiple functions in its therapeutic potential for the induction and maintenance of T cell response, the exact role of PI3K-Akt in IL-15 increased adaptive immunity is still poorly understood. In this study, we demonstrated that ex vivo IL-15 administration increased BMPC capability of antigen uptake and the expression of costimulatory molecules (such as CD80 and 4-1BB(CD137) ligand [4-1BBL]) and MHC class I molecule via PI3K-Akt pathway. Importantly, PI3K-Akt activity was not only necessary for IL-15 augmented BMPC cross-presentation and CTL priming, but also facilitated IL-15 increased therapeutic potential of the cytolytic capacity and maintenance of BMPC-activated T cells. Thus, these data suggested that PI3K-Akt activity contribute to the superior immune function of IL-15-administrated BMPC and thereby might be therapeutic potential for adaptive immunity.

Inflammatory bowel disease- and Barrett's esophagus-associated neoplasia: the old, the new, and the persistent struggles.

Early diagnosis of and adequate therapy for premalignant lesions in patients with inflammatory bowel disease (IBD) and Barrett's esophagus (BE) has been shown to decrease mortality. Endoscopic examination with histologic evaluation of random and targeted biopsies remains the gold standard for early detection and adequate treatment of neoplasia in both these diseases. Although eventual patient management (including surveillance and treatment) depends upon a precise histologic assessment of the initial biopsy, accurately diagnosing and grading IBD- and BE-associated dysplasia is still considered challenging by many general as well as subspecialized pathologists. Additionally, there are continuing updates in the literature regarding the diagnosis, surveillance, and treatment of these disease entities. This comprehensive review discusses the cancer risk, detailed histopathological features, diagnostic challenges, and updates as well as the latest surveillance and treatment recommendations in IBD- and BE-associated dysplasia.

Prevalence of incontinence among cognitively impaired older residents in long-term care facilities in East Asia: A cross-sectional study.

AIM: The current study was an international cross-sectional study comparing the prevalence of incontinence among cognitively impaired older residents in long-term care facilities in East Asia, including Japan, Korea, China, Taiwan and Thailand between 2015 and 2016. METHODS: Participants were cognitively impaired older residents in long-term care facilities. Demographic data were collected. The Clinical Dementia Rating scale was used to assess dementia severity, and the Barthel Index was used as a surrogate measure of incontinence and toilet use dependence. The prevalence of urinary incontinence and fecal incontinence were examined. Multiple logistic regression analysis was used to predict incontinence and toilet use dependence. RESULTS: We analyzed data from 662 participants (age 82.6 ± 9.9 years, 57.6% women). The prevalence of urinary incontinence ranged from 10.1% in Taiwan to 71.0% in Korea. The prevalence of fecal incontinence varied from 4.0% in Taiwan to 57.0% in Korea. A higher Clinical Dementia Rating score was a significant predictor of urinary and fecal incontinence and toilet use dependence (P < 0.0001). CONCLUSIONS: The current survey showed a high prevalence of incontinence in long-term care residents in East Asia, and identified challenges for future studies. Development of clinical guidelines for incontinence care in cognitively impaired older persons is urgently required. Geriatr Gerontol Int 2019; 19: 444-450.

Variations and factors associated with psychotropic use in cognitively impaired elderly residing in long-term care facilities in East Asia: a cross-sectional study.

BACKGROUND: The aim of this cross-sectional study was to compare patterns of psychotropic prescription drug use among cognitively impaired residents in long-term care facilities in East Asia and to explore factors associated with these patterns. METHODS: This study included elderly participants with cognitive impairments residing in long-term care facilities with and without dementia care units in Japan, South Korea, China, Taiwan, and Thailand. The Mini-Mental State Examination, the Clinical Dementia Rating, and the Neuropsychiatric Inventory, Nursing Home version were used to assess cognitive status, examine dementia severity, and evaluate behavioural psychological symptoms of dementia, respectively. The rate of psychotropic drug use and the relationship between the number of psychotropic drugs and clinical factors were examined. RESULTS: In total, 662 people were analyzed. Facilities with dementia care units had a higher rate of anti-dementia drug use than regular elderly care sites. Among the three dementia care sites, a Japanese hospital and a Korean site had a high rate of antipsychotic use and use of other types of psychotropics, whereas these drugs were used at a low rate in a Chinese nursing home. Patterns of psychotropic drug use may be partially associated with local regulations and facility type. Poly-pharmacy was identified as a common problem at all study sites. CONCLUSIONS: Our findings will be beneficial for health-care professionals and policymakers when developing practice guidelines and strategies to regulate overuse of psychotropics and poly-pharmacy. Prospective studies are needed to examine patterns of psychotropic prescriptions and to promote evidence-based practice.

Prevalence of behavioural and psychological symptoms of dementia in cognitively impaired elderly residents of long-term care facilities in East Asia: a cross-sectional study.

OBJECTIVE: The aim of this study was to compare the prevalence of behavioural and psychological symptoms of dementia (BPSD) in cognitively impaired elderly residents of long-term care facilities in East Asia and to explore the factors associated with these patterns. METHODS: This was a cross-sectional survey of BPSD in cognitively impaired elderly residents of long-term care facilities in Japan, South Korea, China, Taiwan, and Thailand. The Mini-Mental State Examination, Clinical Dementia Rating (CDR), and Neuropsychiatric Inventory, Nursing Home version (NPI-NH), were used to assess cognitive status, dementia severity, and BPSD, respectively. NPI-NH subscale severity scores were multiplied by frequency scores to obtain the subscale scores and aggregated into two groups based on score (clinically insignificant = 1- 3; clinically significant ≥4). RESULTS: Data from 662 people were analyzed. Median age, median Mini-Mental State Examination scores, and median CDR scores differed significantly among the seven study sites. The prevalence of BPSD varied from 64% in Taiwan to 100% in dementia care units in Japan, and the median total NPI-NH scores ranged from 2 in Taiwan to 14 in dementia care units in Japan. After stratification of the sample by dementia severity and clinical significance of NPI-NH scores, differences in the prevalence of clinically significant BPSD were mostly observed among facilities dedicated to dementia patients in the CDR 1 group. In the CDR 3 group, the prevalence of some clinically significant BPSD, such as apathy, was high even among study sites with low median total NPI-NH scores. CONCLUSIONS: Our findings may suggest referral and selection biases in the study sites. Future prospective studies are needed to address the impact of environmental and care factors on the occurrence of BPSD in Asian countries.

A meta-analysis of serum cancer antigen 125 array for diagnosis of ovarian cancer in Chinese.

OBJECTIVE: To further evaluation the diagnosis accuracy of serum cancer antigen 125 (CA125) in the diagnosis of ovarian cancer in Chinese patients. MATERIALS AND METHODS: The PubMed, Wanfang and CNKI databases were electric searched and relevant diagnosis trials were reviewed and finally included in this meta-analysis. The diagnosis sensitivity, specificity, positive likely hood ratio (+LR), negative likely hood ratio (-LR), diagnostic odds ratio (DOR) and receiver operating characteristic curve were pooled by Meta DiSc 1.4 software. RESULTS: Nineteen studies with a total of 2426 subjects were included in this meta-analysis. The pooled sensitivity, specificity, +LR, -LR and DOR were 0.75 (95% confidence interval = 0.73-0.78), 0.80 (0.77-0.82), 4.52 (3.27-6.26), 0.31 (0.28-0.35) and 15.76 (10.45-23.75) respectively. The area under the summary receiver operating characteristic curve was 0.84. CONCLUSION: Serum CA125 was potential biomarker for diagnosis of ovarian cancer with acceptable diagnosis value.

Changes in activity patterns after the oral administration of brotizolam in institutionalized elderly patients with dementia.

BACKGROUND: Little is known about the side effects of sedative-hypnotic agents in elderly dementia patients with sleep disorders. The present study describes activity pattern changes after a single dose of brotizolam in elderly patients with dementia. METHODS: We conducted retrospective analysis of prospectively collected data from a case series at Asakayama Hospital (Osaka, Japan) between September 2008 and September 2009. Around-the-clock movements of dementia patients who were administered a single dose of brotizolam were recorded by the integrated circuit tag monitoring system during a 4-week baseline and 7-day peri-administration period. Diurnal and nocturnal activity levels and the onset times of the least-active and most-active phases were then measured. RESULTS: Seven patients (four men, three women; age range 59-85 years) were analyzed. All seven patients had disturbed activity patterns during the peri-administration period. Compared with the pre-administration period, the incidence of reversed rest-activity pattern increased significantly in the post-administration period, as measured by the distance moved per hour (P < 0.000). Patients with advanced stages of dementia had prolonged and delayed activity responses. CONCLUSIONS: Findings showed changes in activity levels and reversed active/resting phases after a single dose of brotizolam in elderly patients with dementia. Use of brotizolam in elderly patients with dementia, especially in advanced stages, calls for closer attention and longer observation periods.

[Difference in sundowning of wandering behavior in patients with Alzheimer disease and frontotemporal dementia].

OBJECTIVES: To delineate the difference in sundowning of wandering behavior between patient with Alzheimer Disease (AD) and frontotemporal dementia (FTD). METHODS: The study was conducted in a dementia care unit at A hospital in Osaka, Japan from September 2008 to September 2009. Twenty-four-hour movements of 27 ambulatory inpatients with AD and 7 with FTD were coded consecutively by the IC tag monitoring system. RESULTS: Repeated measures ANOVA after the adjustment of Huynh-Feldt Epsilon (H-F) showed no significant difference in 24 h standardized activity level between two groups (F = 3.74, P = 0.06), and there was no interaction between diagnosis and time (F = 1.42, P > 0.05). The standardized activity levels gradually increased from late afternoon to evening and then reached the highest point at 18:00 in AD group and 19:00 in FTD group. Test of within-subjects contrasts for order 17 was significant (F = 5.24, P < 0.05) and for order 9 was a tendency of significant (F = 4.26; P = 0.05) between two groups. AD group was significant greater active at 5:00, 6:00 and 7:00 (0.75 ± 0.08 vs 0.35 ± 0.16, F = 4.91; 1.13 ± 0.13 vs 0.49 ± 0.26, F = 5.06; 1.24 ± 0.15 vs 0.56 ± 0.28, F = 4.47 respectively, P < 0.05), and less active at 16:00 (1.65 ± 0.11 vs 2.22 ± 0.22, P < 0.05) comparing to FTD group by Bonferroni's multiple comparison test. Meanwhile, the time of peak value of hourly distance moved per day (PV-time) was delayed in FTD group comparing to AD by circular χ² test (14:12 ± 5:12 vs 15:47 ± 4:19, χ² = 87.31, P < 0.01). CONCLUSIONS: The study suggests great possibility of sundowning of wandering behavior in both two subtypes with different temporal pattern of wandering behavior. Comparing to FTD patients, AD patients showed an advanced PV-time and prolonged active phase.

A novel covalent coupling method for coating of capillaries with liposomes in capillary electrophoresis.

A novel covalent coupling method for coating of capillaries with liposomes has been developed, which includes three steps: (i) epoxy-diol coating, (ii) activation with 2,2,2-trifluoroethanesulfonyl chloride, and (iii) liposome coupling. The coating conditions, such as the reaction time and temperature of liposome coupling, the content of dimyristoylphosphatidylethanolamine in liposomes, were optimized. Vesicles were visualized on the inner silica wall as confirmed by atomic force microscopy. The effectiveness of the coating was demonstrated by investigating the effect of pH of BGE on EOF and separating neutral compounds. The intra- and inter-capillary variations in EOF are 4.02% RSD (n=30) and 6.72% RSD (n=4) respectively, and the coated capillaries can be used to perform analysis at least for one month without any performance deterioration when stored at 4 degrees C. A set of drugs with diverse structures was applied into the developed liposome-coated CE. The normalized capacity factor (K) was introduced to quantitatively evaluate drug-membrane interactions. The relationship between log K and the fraction dose absorbed in humans (Fa%) shows that the liposome-coated CE can be utilized for in vitro prediction of Fa% of drugs that follow the transcellular passive transport route.