SIRT3 facilitates mitochondrial structural repair and functional recovery in rats after ischemic stroke by promoting OPA1 expression and activity.

BACKGROUND: Optical atrophy 1 (OPA1), a protein accountable for mitochondrial fusion, facilitates the restoration of mitochondrial structure and function following cerebral ischemia/reperfusion (I/R) injury. The OPA1-conferred mitochondrial protection involves its expression and activity, which can be improved by SIRT3 in non-cerebral ischemia. Nevertheless, it remains obscure whether SIRT3 enhances the expression and activity of OPA1 after cerebral I/R injury. METHODS: Mature male Sprague Dawley rats were intracranially injected with adeno-associated viral-Sirtuin-3(AAV-SIRT3) and AAV-sh_OPA1, followed by a 90-min temporary blockage of the middle cerebral artery and subsequent restoration of blood flow. Cultured cortical neurons of rats were transfected with LV-SIRT3 or LV-sh_OPA1 before a 2-h oxygen-glucose deprivation and reoxygenation. The rats and neurons were subsequently treated with a selective OPA1 activity inhibitor (MYLS22). The interaction between SIRT3 and OPA1 was assessed by molecular dynamics simulation technology and co-immunoprecipitation. The expression, function, and specific protective mechanism of SIRT3 were examined by various analyses. RESULTS: SIRT3 interacted with OPA1 in the rat cerebral cortex before and after cerebral I/R. After cerebral I/R damage, SIRT3 upregulation increased the OPA1 expression, which enhanced deacetylation and OPA1 activity, thus alleviating cerebral infarct volume, neuronal apoptosis, oxidative pressure, and impairment in mitochondrial energy production; SIRT3 upregulation also improved neuromotor performance, repaired mitochondrial ultrastructure and membrane composition, and promoted the mitochondrial biogenesis. These neuroprotective effects were partly reversed by OPA1 expression interference and OPA1 activity inhibitor MYLS22. CONCLUSION: In rats, SIRT3 enhances the expression and activity of OPA1, facilitating the repair of mitochondrial structure and functional recovery following cerebral I/R injury. These findings highlight that regulating SIRT3 may be a promising therapeutic strategy for ischemic stroke.

SIRT1 restores mitochondrial structure and function in rats by activating SIRT3 after cerebral ischemia/reperfusion injury.

Mitochondrial dysfunction contributes to cerebral ischemia-reperfusion (CI/R) injury, which can be ameliorated by Sirtuin-3 (SIRT3). Under stress conditions, the SIRT3-promoted mitochondrial functional recovery depends on both its activity and expression. However, the approach to enhance SIRT3 activity after CI/R injury remains unelucidated. In this study, Sprague-Dawley (SD) rats were intracranially injected with either adeno-associated viral Sirtuin-1 (AAV-SIRT1) or AAV-sh_SIRT1 before undergoing transient middle cerebral artery occlusion (tMCAO). Primary cortical neurons were cultured and transfected with lentiviral SIRT1 (LV-SIRT1) and LV-sh_SIRT1 respectively before oxygen-glucose deprivation/reoxygenation (OGD/R). Afterwards, rats and neurons were respectively treated with a selective SIRT3 inhibitor, 3-(1H-1,2,3-triazol-4-yl) pyridine (3-TYP). The expression, function, and related mechanism of SIRT1 were investigated by Western Blot, flow cytometry, immunofluorescence staining, etc. After CI/R injury, SIRT1 expression decreased in vivo and in vitro. The simulation and immune-analyses reported strong interaction between SIRT1 and SIRT3 in the cerebral mitochondria before and after CI/R. SIRT1 overexpression enhanced SIRT3 activity by increasing the deacetylation of SIRT3, which ameliorated CI/R-induced cerebral infarction, neuronal apoptosis, oxidative stress, neurological and motor dysfunction, and mitochondrial respiratory chain dysfunction, promoted mitochondrial biogenesis, and retained mitochondrial integrity and mitochondrial morphology. Meanwhile, SIRT1 overexpression alleviated OGD/R-induced neuronal death and mitochondrial bioenergetic deficits. These effects were reversed by AAV-sh_SIRT1 and the neuroprotective effects of SIRT1 were partially offset by 3-TYP. These results suggest that SIRT1 restores the structure and function of mitochondria by activating SIRT3, offering neuroprotection against CI/R injury, which signifies a potential approach for the clinical management of cerebral ischemia.

The relationship between daytime napping and glycemic control in people with type 2 diabetes.

Aim: To examine the association between napping characteristics and glycemic control in people with type 2 diabetes. Design: This study used a cross-sectional design. Methods: A convenience sample of people with type 2 diabetes (N=226) were included. Glycemic control was indicated by HbA1c which was measured by A1C Now®+. Napping characteristics including napping frequency, duration, timing, and type were measured by validated questionnaires. Other variables, such as insomnia, cognitive impairment, and depression were measured by the Insomnia Severity Index, Montreal Cognitive Assessment, and Patient Health Questionnaire-9, respectively. Multivariate linear regression analyses were performed. Results: The sample consisted of 122 women (54.0%), with a median age of 67 years. Their median HbA1c was 6.8%. No significant relationship was found between napping frequency and HbA1c. Among nappers, after controlling for covariates, long napping duration (≥60 min) and morning napping were both associated with poorer glycemic control. Compared with appetitive napping, restorative napping was associated with better glycemic control. Conclusion: Daytime napping (e.g., duration and type) is an important modifiable factor for glycemic control in people with type 2 diabetes. This study provides new insights into the relationship between napping and glucose management among people with diabetes.

Sleep health predicted glucose metabolism among pregnant women: A prospective cohort study.

AIMS: To examine whether sleep health in the first trimester could predict glucose metabolism in the second trimester. METHODS: Pregnant women (N = 127) during the first trimester were recruited (August 2022 to March 2023). Overall sleep health was assessed by the Sleep Health Index. Various dimensions of sleep health were measured using a 7-day sleep diary and questionnaires. The outcomes, including diagnosis of gestational diabetes mellitus (GDM) and HbA1c, were obtained from the medical records in the second trimester. Poisson regression analysis and multiple linear regression were used for data analysis. RESULTS: The average age of the participants was 32.6 years. The incidence of GDM was 28.3 % and the mean HbA1c was 5.2 % (33 mmol/mol). Sleep duration regularity (RR = 1.808; 95 %CI 1.023, 3.196) was associated with GDM after controlling for confounders. SHI total score (ß = -0.278; 95 %CI -0.022, -0.005) and sleep duration regularity (ß = 0.243; 95 %CI 0.057, 0.372) were associated with HbA1c. CONCLUSIONS: Worse sleep health, particularly lower sleep regularity, predicted worse glucose metabolism among pregnant women. Healthcare professional may consider adding sleep-related assessment to prenatal care. Maintaining regular sleep should be encouraged. Studies examining the impact of sleep intervention on glucose metabolism among pregnant women are warranted.

Sleep behaviors predicted sleep disturbances among Chinese health science students: a cross-sectional study.

PURPOSE: Healthy sleep is essential for individuals' physiological and psychological health. Health science students experience a high prevalence of sleep disturbances which may be due to maladaptive behaviors. This study aimed to examine the associations of sleep behaviors including sleep hygiene and bedtime procrastination with the associations of sleep disturbances (e.g., poor sleep quality, insomnia, and short sleep). METHODS: This cross-sectional study included health science students from a medical university in Shanghai, China. Sleep disturbances included poor sleep quality, insomnia, and short sleep. They were measured by the Pittsburgh Sleep Quality Index (PSQI), Insomnia Severity Index (ISI), and one question "How many hours of sleep did you usually get during the past week?", respectively. Sleep behaviors included sleep hygiene and bedtime procrastination measured by the Sleep Hygiene Index (SHI) and Bedtime Procrastination Scale (BPS), respectively. Logistic regression was performed while controlling for potential confounders. RESULTS: A total of 464 health science students participated. Poorer overall sleep hygiene and more bedtime procrastination were independently associated with higher odds of poor sleep quality (OR=1.065, 95% CI 1.028-1.103; OR=1.040, 95% CI 1.006-1.075, respectively) and insomnia (OR=1.059, 95% CI 1.018-1.101; OR=1.093, 95% CI 1.049-1.139, respectively). More bedtime procrastination was associated with higher odds of short sleep (OR=1.148, 95% CI 1.093-1.206). Commonly reported specific sleep behaviors, such as "Going to bed later than intended", "Doing other things than sleep at bedtime", and "Easily stopping what I am doing at bedtime", were also related to higher odds of sleep disturbances. CONCLUSIONS: Sleep hygiene and bedtime procrastination were strong predictors of sleep disturbances. Tailored interventions targeting specific sleep behaviors are warranted to clarify their effect on sleep disturbances.

Information Needs and Its Association With Depressive Symptoms in People With Type 2 Diabetes.

PURPOSE: The purpose of this study was to describe the information needs and examine its association with depressive symptoms in people with type 2 diabetes (T2D). METHODS: A descriptive, correlational design was used. People with T2D (N = 358) were recruited from 12 communities in Shanghai, China. Self-reported information needs and depressive symptoms were measured using the Information Needs in Diabetes Questionnaire and Patient Health Questionnaire-9 (PHQ-9), respectively. Multivariate linear regression analysis was performed. RESULTS: The participants were 64.8 years on average, and 46.6% were men. One hundred fifty-one (42.2%) had depressive symptoms (PHQ-9 ≥ 5). Participants had the least knowledge about "diabetes research," "acute complications," and "lifestyle adjustment." The sample had the highest levels of information needs about topics including "mental strain," "treatment/therapy," and "diabetes in everyday life." Compared to those without depressive symptoms, those experiencing depressive symptoms were less informed and had higher levels of need for further information. Controlling for covariates, higher levels of need for further information were significantly associated with greater depressive symptoms (B = 0.368, 95% CI, 0.155-0.582, P = .001). CONCLUSIONS: This study demonstrated areas that should be prioritized when meeting patients' information needs. It also showed the potential negative effect of unmet information needs on depression. These findings may help develop a more tailored intervention for people with T2D.

Effect of Temozolomide Combined with Intensity Modulated Radiation Therapy on Serum Factor, Immune Function and Clinical Efficacy in Postoperative Glioma Patients.

To investigate the effect of Temozolomide combined with intensity modulated radiation therapy on serum factor, immune function and clinical efficacy in postoperative glioma patients. One hundred twenty-four patients with high-grade glioma admitted to the First Affiliated Hospital of Zhengzhou University were selected and randomly divided into the study group and the control group, with 62 cases in each group. The control group was given intensity modulated radiation therapy alone, and the study group was given Temozolomide combined with intensity modulated radiation therapy. The clinical efficacy, serum factor, immune function and adverse reactions were observed and compared. The overall response rate of the study group was 95.16%, which is higher than 83.87% in the control group, and the differences were significant (P < 0.05); After the treatment, the serum VEGF, EGF and HGF indicators and diverse immune function indicators were superior to those in the control group, and the differences indicated significance (P < 0.05); the incidence of adverse reactions in the study group was 37.10%, which is higher than 25.81% in the control group, but the differences showed no significance (P > 0.05). Temozolomide combined with intensity modulated radiation therapy could improve the level of serum factor in postoperative glioma patients, strengthen the immune function of the patients, and effectively facilitate the clinical comprehensive efficacy without increasing adverse reactions.

Neuropilin-1 promotes mitochondrial structural repair and functional recovery in rats with cerebral ischemia.

OBJECTIVES: Available literature documents that ischemic stroke can disrupt the morphology and function of mitochondria and that the latter in other disease models can be preserved by neuropilin-1 (NRP-1) via oxidative stress suppression. However, whether NRP-1 can repair mitochondrial structure and promote functional recovery after cerebral ischemia is still unknown. This study tackled this very issue and explored the underlying mechanism. METHODS: Adeno-associated viral (AAV)-NRP-1 was stereotaxically inoculated into the cortex and ipsilateral striatum posterior of adult male Sprague-Dawley (SD) rats before a 90-min transient middle cerebral artery occlusion (tMCAO) and subsequent reperfusion. Lentivirus (LV)-NRP-1 was transfected into rat primary cortical neuronal cultures before a 2-h oxygen-glucose deprivation and reoxygenation (OGD/R) injury to neurons. The expression and function of NRP-1 and its specific protective mechanism were investigated by Western Blot, immunofluorescence staining, flow cytometry, magnetic resonance imaging, transmission electron microscopy, etc. The binding was detected by molecular docking and molecular dynamics simulation. RESULTS: Both in vitro and in vivo models of cerebral ischemia/reperfusion (I/R) injury presented a sharp increase in NRP-1 expression. The expression of AAV-NRP-1 markedly ameliorated the cerebral I/R-induced damage to the motor function and restored the mitochondrial morphology. The expression of LV-NRP-1 alleviated mitochondrial oxidative stress and bioenergetic deficits. AAV-NRP-1 and LV-NRP-1 treatments increased the wingless integration (Wnt)-associated signals and ß-catenin nuclear localization. The protective effects of NRP-1 were reversed by the administration of XAV-939. CONCLUSIONS: NRP-1 can produce neuroprotective effects against I/R injury to the brain by activating the Wnt/ß-catenin signaling pathway and promoting mitochondrial structural repair and functional recovery, which may serve as a promising candidate target in treating ischemic stroke.

Psychometric evaluation of the Chinese version of Sleep Health Index in pregnant women.

OBJECTIVE: The aim of this study was to investigate the psychometric properties of the Chinese version of the Sleep Health Index (SHI-C) among pregnant women. DESIGN: Cross-sectional design. SETTING: Outpatient clinic of three hospitals in China. PARTICIPANTS: Pregnant women (N = 264) aged between 18 and 45 years were recruited via convenience sampling. METHODS: The Pittsburgh Sleep Quality Index (PSQI), Epworth Sleepiness Scale (ESS) and Insomnia Severity Index (ISI) were used to measure sleep quality, daytime sleepiness, and insomnia, respectively. The Fatigue Assessment Scale (FAS) and the Edinburgh Postnatal Depression Scale (EPDS) were used to measure fatigue and depression, respectively. Structural validity was assessed via confirmatory factor analysis (CFA). Concurrent and convergent validity were assessed using bivariate correlation analyses. Known-group validity was assessed by comparing the SHI-C score between different groups. Cronbach's α was calculated for reliability. FINDINGS: The average sample age was 30.6 years old and their average score of SHI-C was 86.4 (SD 8.2). Based on PSQI, ISI, and ESS, 43.6%, 32.2%, and 26.9% had poor sleep quality, insomnia, and excessive daytime sleepiness, respectively. The SHI-C total and sleep quality sub-index scores had moderate to strong correlations with both PSQI (r=-0.542, p<0.01; r=-0.648, p<0.01) and ISI (r=-0.692, p<0.01; r=-0.752, p<0.01). The SHI-C total and sleep quality sub-index scores were significantly associated with ESS, FAS, and EPDS (r=-0.171 to -0.276; p<0.01). The SHI-C total score was higher in the second trimester and among those who were working, never drank coffee, or took a nap every day. The Cronbach's α of the SHI-C total and the sleep quality sub-index were 0.723 and 0.806, respectively. The Cronbach's α of sleep duration and disordered sleep sub-indices were 0.594 and 0.545, respectively. KEY CONCLUSIONS: Overall, the SHI-C has good validity and acceptable reliability among the pregnant population in China. It can be a useful tool for the assessment of sleep health. More research is warranted to refine the sleep duration and disordered sleep sub-indices. IMPLICATIONS FOR PRACTICE: The use of SHI-C would facilitate the assessment of sleep health among pregnant women, which could contribute to the promotion of perinatal care.

Cross-cultural adaptation and validation of the Chinese version of the Sleep Health Index.

OBJECTIVE: To generate the Chinese Sleep Health Index (SHI-C) in Mandarin with cross-cultural adaptations and test its psychometric properties. METHODS: This study used a cross-sectional design. Health science students were included (N = 271) and a sub-set (n = 74) was invited for the re-test. Cross-cultural adaptation of the SHI-C was performed prior to formal validation. The SHI-C, Pittsburgh Sleep Quality Index (PSQI), Insomnia Severity Index (ISI), Bedtime Procrastination Scale, and Sleep Hygiene Index were used to measure variables of interest. Exploratory factor analysis was used to evaluate the structure validity. Bivariate analyses were used to evaluate the construct validity. RESULTS: Exploratory factor analysis identified 3 factors (ie, sleep quality, sleep duration, and disordered sleep) accounting for 55.6% of the total variance. The SHI-C total and sleep quality sub-index scores were significantly associated with both PSQI global score (r = -0.132, p < .05; r = -0.182, p < .01, respectively) and ISI score (r = -0.655, p < .05; r = -0.820, p < .05, respectively). SHI-C total, sleep quality sub-index, and sleep duration sub-index scores were significantly associated with Bedtime Procrastination Scale and Sleep Hygiene Index scores (r = -0.238 to -0.368, p < .05). Students with insomnia (ISI > 9) or poor sleep quality (PSQI > 5) had significantly lower SHI-C scores than those without (73.5 vs. 89.0, p < .01; 84.1 vs. 86.7, p < .05, respectively). SHI-C showed good internal consistency (Cronbach's alpha = 0.73) and test-retest reliability (intraclass correlation coefficient = 0.82). CONCLUSIONS: The SHI-C demonstrated good validity and adequate reliability in a Chinese sample of health science students. It could be used to measure sleep health in future research and practice. Psychometric properties of the SHI-C among other Chinese populations remain to be confirmed.

3'UTR of SARS-CoV-2 spike gene hijack host miR-296 or miR-520h to disturb cell proliferation and cytokine signaling.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has becoming globally public health threat. Recently studies were focus on SARS-CoV-2 RNA to design vaccine and drugs. It was demonstrated that virus RNA could play as sponge to host noncoding RNAs to regulate cellular processes. Bioinformatic research predicted a series of motif on SARS-CoV-2 genome where are targets of human miRNAs. In this study, we used dual-luciferase reporter assays to validate the interaction between 3'UTR of SARS-CoV-2 S (S-3'UTR) gene and bioinformatic predicted targeting miRNAs. The growth of 293T cells and HUVECs with overexpressed S-3'UTR was determined, while miRNAs and IL6, TNF-α levels were checked in this condition. Then, miR-296 and miR-602 mimic were introduced into 293T cells and HUVECs with overexpressed S-3'UTR, respectively, to reveal the underlying regulation mechanism. In results, we screened 19 miRNAs targeting the S-3'UTR, including miR-296 and miR-602. In 293T cell, S-3'UTR could inhibit 293T cell growth through down-regulation of miR-296. By reducing miR-602, S-3'UTR could induce HUVECs cell proliferation, alter the cell cycle, reduce apoptosis, and enhanced IL6 and TNF-αlevel. In conclusion, SARS-CoV-2 RNA could play as sponge of host miRNA to disturb cell growth and cytokine signaling. It suggests an important clue for designing COVID-19 drug and vaccine.

Effect of circPUM1 on radioresistance of cervical cancer cells through targeting miR-144-3p.

OBJECTIVE: To investigate the effect of circular RNA pumilio RNA binding family member (circPUM) 1 on radioresistance of cervical cancer cells and its mechanism. METHODS: Cancer tissue and corresponding paricancerous tissue samples were collected from 47 patients with cervical cancer who underwent surgical treatment in the Second Affiliated Hospital of Zhengzhou University from August 2019 to February 2020. The expression levels of circPUM1 and miR-144-3p in cervical cancer tissues and paricancerous tissues were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The Pearson method was used to analyze the correlation between circPUM1 and miR-144-3p expression in cervical cancer tissues. circPUM1 lentiviral short hairpin RNA (sh-circPUM1) and its negative control (sh-NC), miR-144-3p oligonucleotide mimic (miR-144-3p mimic) and its negative control (miR-NC), sh-circPUM1 and miR-144-3p inhibitor (anti-miR), and sh-circPUM1 and anti-miR negative control (anti-miR-NC) were transfected into human cervical carcinoma SiHa cells, respectively, and the cells were irradiated with 0 and 4 Gy irradiation doses. Cell proliferation, colony formation, apoptosis, migration and invasion were detected by cell counting kit (CCK-8 method), plate colony formation assay, flow cytometry and Transwell assay, respectively. The protein expression of cleaved-caspase3 was detected by Western blotting. The targeting relationship between circPUM1 and miR-144-3p was analyzed with Starbase platform. RESULTS: Compared with adjacent tissue, the expression of circPUM1 in cervical cancer tissue was significantly increased ( P<0.05), while the expression of miR-144-3p was decreased ( P<0.05). The circPUM1 was negatively correlated with miR-144-3p ( r=-0.9282, P<0.01). After transfection with sh-circPUM1 or miR-144-3p mimic, the inhibition rate of cell proliferation, the rate of apoptosis and the expression level of cleaved-caspase3 protein increased (all P<0.05), while the number of colonies formed, migrated and invaded cells decreased (all P<0.05). CircPUM1 could targeted to miR-144-3p. After co-transfection of sh-circPUM1 and anti-miR, the inhibition rate of cell proliferation, the rate of apoptosis and the expression level of cleaved-caspase3 protein significantly decreased (all P<0.05), while the number of colonies formed, migrated and invaded cells increased (all P<0.05). CONCLUSION: Silencing circPUM1 may inhibit the proliferation, colony formation, migration, invasion and induce apoptosis of cervical cancer cells through targeting and regulating the expression of miR-144-3p.

Associations between sleep variability and cardiometabolic health: A systematic review.

This review explored the associations between sleep variability and cardiometabolic health. It was performed following PRISMA guidelines. We identified 63 studies. Forty-one studies examined the association between sleep variability and body composition, with 29 examined body mass index (BMI). Thirteen studies used social jet lag (SJL), n = 30,519, with nine reporting a null association. Eight studies used variability in sleep duration (n = 33,029), with five reporting a correlation with BMI. Fourteen studies (n = 133,403) focused on overweight/obesity; significant associations with sleep variability were found in 11 (n = 120,168). Sleep variability was associated with weight gain (seven studies; n = 79,522). Twenty-three studies examined glucose outcomes. The association with hemoglobin A1c (16 studies, n = 11,755) differed depending on populations, while associations with diabetes or glucose were mixed, and none were seen with insulin resistance (five studies; n = 6416). Sixteen studies examined cardiovascular-related outcomes, with inconsistent results. Overall significant associations were found in five studies focusing on metabolic syndrome (n = 7413). In summary, sleep variability was likely associated with obesity, weight gain, and metabolic syndrome. It might be associated with hemoglobin A1c in people with type 1 diabetes. The associations with other outcomes were mixed. This review highlighted the possible association between sleep variability and cardiometabolic health.

Characteristics of tinnitus and factors influencing its severity.

Background and Objectives: There is a wealth of information regarding the treatment methods for tinnitus; however, the treatment available is unsatisfactory because of the following reasons: first, tinnitus has various etiologies and second, it has distinct heterogeneity among different individuals. Numerous studies have focused on understanding the causes of tinnitus severity, but the conclusions have been inconsistent. The purpose of the present study was to define factors that differentially influence subjectively perceived tinnitus severity. Methods: Clinical data of patients with chronic tinnitus who visited our outpatient clinic from April 2020 to April 2021 were collected. Tinnitus Handicap Inventory (THI) and Tinnitus Evaluation Questionnaire (TEQ) were used to evaluate tinnitus severity among patients, and the independent factors influencing the severity of tinnitus were investigated by performing univariate and multivariate stepwise regression analyses. Results: Eleven variables were associated with THI and TEQ scores, of which nine were identical. Multiple regression analyses results revealed that five variables had a significantly unique predictive effect on tinnitus severity based on THI and the TEQ scores. Three factors including Self-Rating Scale of Sleep (SRSS), change in loudness, and Self-Rating Anxiety Scale (SAS) were identical. Conclusion: Sleep status, anxiety level, and change in loudness in patients with chronic tinnitus were significantly correlated with severity of tinnitus. Follow-up studies should investigate the causal relationship between these factors and tinnitus severity.

Circ_0060055 Promotes the Growth, Invasion, and Radioresistance of Glioblastoma by Targeting MiR-197-3p/API5 Axis.

Circular RNA (circRNA) has been shown to be involved in the regulation of human disease progression. Our study aims to reveal the role of circ_0060055 in the progression of glioblastoma (GBM) and its potential molecular mechanism. The expression of circ_0060055, microRNA (miR)-197-3p, and apoptosis inhibitor 5 (API5) was determined by quantitative real-time PCR. GBM cell proliferation, apoptosis, and invasion were assessed using cell counting kit 8 assay, colony formation assay, EdU assay, flow cytometry, and transwell assay. Besides, the radiosensitivity of cells also was assessed using colony formation assay. The interaction between miR-197-3p and circ_0060055 or API5 was analyzed by dual-luciferase reporter assay and RNA pull-down assay. Animal experiments were conducted to measure the effect of circ_0060055 on GBM tumor growth and radiosensitivity in vivo. Circ_0060055 was overexpressed in GBM tumor tissues and cells, and its silencing suppressed GBM cell proliferation and invasion, while promoted apoptosis and radiosensitivity. In terms of mechanism, circ_0060055 could interact with miR-197-3p, and miR-197-3p could target API5. API5 expression also could be positively regulated by circ_0060055. Function experiments suggested that miR-197-3p inhibitor abolished the effect of circ_0060055 knockdown on GBM cell growth, invasion, and radiosensitivity. MiR-197-3p repressed GBM cell progression and improved radiosensitivity, and this effect was eliminated by API5 upregulation. In vivo experiments confirmed that circ_0060055 knockdown reduced GBM tumor growth and enhanced the radiosensitivity of tumors. This study revealed that circ_0060055 contributed to GBM progression and radioresistance through miR-197-3p/API5 pathway, providing a potential target for GBM treatment.

HECTD3 enhances cell radiation resistance and migration by regulating LKB1 mediated ZEB1 in glioma.

Homologous to the E6-associated protein carboxyl terminus domain containing 3 (HECTD3) has been reported to play a role in carcinogenesis. Here, we explored the role of HECTD3 in regulating the radiation resistance of glioma, and the underlying mechanism. HECTD3 expressions in glioma tissues were assessed using Western blotting, quantitative reverse transcription (qRT)-polymerase chain reaction (PCR) and immunohistochemistry. Glioma cells were exposed to 2-, 4-, 6- or 8-Gy X-ray to mimic the radiation treatment. Cell count kit-8 (CCK-8), clone formation assay, flow cytometry assay, transwell chambers and animal assay were used to test cell viability, apoptosis, migration, invasiveness and tumourigenesis, respectively. HECTD3 expression was increased in glioma tissues, especially from patients with radiation resistance. Knockdown of HECTD3 promoted cell apoptosis and inhibited cell viability under the condition of 8-Gy X-ray, as well as suppressed cell migration and invasiveness. In mechanism, HECTD3 positively regulated ZEB1 (zinc finger E-box binding hemeobox 1) expression through regulating the ubiquitination of liver kinase B1 (LKB1) protein. Overexpression of ZEB1 significantly abolished the effects of HECTD3 downregulation in inhibiting the radiation resistance and migration of glioma cells. Moreover, downregulation of HECTD3 further enhanced the anti-tumour effect of X-ray on glioma growth in vivo. In conclusion, HECTD3 was overexpressed in glioma patients with radiation resistance. Knockdown of HECTD3 sensitized glioma cells to radiation and inhibited cell migration by downregulating ZEB1 expression via regulating the ubiquitination of LKB1 protein. This study reveals that HECTD3 might be a potent target to enhance the radiation sensitivity of glioma.

Identification of a peptide that crosses the blood-cerebrospinal fluid barrier by phage display technology.

Treatments of brain diseases are heavily limited by the existence of the blood-brain barrier (BBB), which precludes efficient drug delivery to the brain. Compared with the BBB, drugs may have a better likelihood of reaching the brain via the cerebrospinal fluid (CSF) because of the lack of a barrier between the CSF and the brain. In this study, phage display technology was effectively applied to screen novel peptides as targeting motifs to transport drugs across the blood-cerebrospinal fluid barrier (BCSFB). We applied a phage seven-mer cyclic peptide library (Ph.D.-C7C™) intravenously to rats and later recovered phages from the CSF. After several rounds of screening, the candidate phages that could cross the BCSFB were enriched. Several bacteriophage clones from the final round were randomly selected and sequenced. A peptide sequence denoted as PMK, which was demonstrated to be able to cross the BCSFB via in vivo optical imaging analysis, could be used in the future for the construction of targeted drug delivery systems.

Low vitamin D levels and frequencies of regulatory T cells (Tregs) are associated with adenotonsillar hypertrophy in children.

OBJECTIVE: To evaluate the levels of vitamin D and the frequencies of regulatory T cells (%Tregs) in children undergoing adenotonsillectomies (T&As) and their controls. METHODS: We prospectively collected data from 130 children aged from 2 to 14 years old undergoing T&As and 60 undergoing unrelated elective procedures from November 1, 2015 to December 20, 2017 at the First Affiliated Hospital of Anhui Medical University. Demographic and disease specific data was obtained in addition to blood samples for the measurement of 25-hydroxy (OH)-vitamin D, interleukin-10 and %Tregs. RESULTS: Among the 130 patients undergoing T&As who had 25(OH) vitamin D levels measured, 40.8% were vitamin D deficient (25(OH) vitamin D < 20 ng/mL), 42.3% were insufficient (20 ng/mL < 25(OH) vitamin D < 30 ng/mL), only 16.9% were sufficient (25(OH) vitamin D > 30 ng/mL). Compared with the control group, children undergoing adenotonsillectomies exhibited a significant decrease in the level of serum 25(OH) vitamin D and %Tregs (p < 0.01, p < 0.01). The level of 25(OH) vitamin D and % Tregs did not correlate to parameters like BMI, age, sex in the children undergoing T&As. The lower Vitamin D levels were related to higher OSA-18 scores (Pearson correlation, r = -0.476, p < 0.01), tonsil size (Spearman rank correlation, r = -0.563)and adenoid size (Spearman rank correlation, r = -0.291). In the different vitamin D concentration groups, the mean values of %Tregs were not equal (ANOVA, F = 7.389, p = 0.001). CONCLUSION: Children undergoing T&As have a lower level of 25(OH) vitamin D and %Tregs. Low 25(OH) vitamin D levels were related to higher OSA-18 scores and greater lymphoid tissue size rather than sex, age, increased BMI. Vitamin D and Treg cells are associated with adenotonsillar hypertrophy.

Experimental Study and Clinical Observation on the Improvement Effect of Lienal Polypeptide on Blood Toxicity and Immune Injury Induced by Radiotherapy.

Aims: To investigate the immune and gastrointestinal functional effects of lienal polypeptide (LP) treatment in tumor-bearing mice and carcinoma patients receiving radiotherapy (RT), and to detect hematological indicators and T lymphocyte subsets. Methods: Tumor-bearing mice were randomly divided into five groups: the control group, the RT group, the RT+LP-L (1.7 mg/kg, low dosage of LP) group, the RT+LP-M (5.2 mg/kg, middle dosage of LP) group, and the RT+LP-H (10.4 mg/kg, high dosage of LP) group. In addition, carcinoma patients were randomly divided into two groups. The observation group was given LP during RT, and the control group was only treated with RT. We then compared the myelosuppression, gastrointestinal reactions, and clinical efficacy among groups. Results: In the animal experiments, compared with the control group, the number of leukocytes and lymphocytes of the mice in the "RT" group decreased (p < 0.05). Animals receiving LP evidenced a dose-response curve with regard to the number of leukocytes and lymphocytes that was proportional to the LP dose, increased (p < 0.05). Flow cytometric analyses showed that LP treatment of the mice increased the numbers of CD3+, and CD4+ T cells and theCD4+/CD8+ ratio. In our clinical study, the Radiation Therapy Oncology Group (RTOG)/European Organization for Research and Treatment of Cancer (EORTC) criteria were used for measuring myelosuppression and gastrointestinal reactions. The RTOG/EORTC grade 3 or 4 inhibition rate of leukocytes, granulocytes, hemoglobin, platelets, and gastrointestinal toxic effects in the observation group were significantly lower than that in the control group (p < 0.05). Conclusion: LP can improve the hematopoietic and immune function of RT-treated mice and reduce the hematological and gastrointestinal toxicity of patients treated with RT and improve the quality of life.

Effects of PLK1 on proliferation, invasion and metastasis of gastric cancer cells through epithelial-mesenchymal transition.

Effects of polo-like kinase (PLK1) on proliferation, migration and invasion capacities of gastric cancer cells through epithelial-mesenchymal transition (EMT) were investigated. Small-interfering ribonucleic acid (siRNA) with targeted interference in PLK1 gene was designed and transfected into gastric cancer MGC-803 cells via Lipofectamine to inhibit the expression of PLK1 gene in MGC-803 cells. The proliferation of MGC-803 cells was detected via methyl thiazolyl tetrazolium (MTT) assay. The mRNA and protein expression of PLK1 and EMT-related marker (E-cadherin) was detected via real-time polymerase chain reaction and western blot analysis, respectively. The effects of interference in PLK1 gene on migration and invasion of MGC-803 cells were studied via wound healing assay and Transwell chamber assay, respectively. Results of MTT assay showed that compared with that in control group, the cell proliferation in PLK1 siRNA group was significantly inhibited (p<0.01). Compared with those in control group, the mRNA and protein expression of PLK1 in PLK1 siRNA group was significantly decreased (p<0.01), but the mRNA and protein expression of E-cadherin was obviously upregulated (p<0.01). Results of wound healing assay and invasion assay showed that the capacity of migration and invasion of MGC-803 cells in PLK1 siRNA group was significantly inhibited compared with those in control group (p<0.01). In conclusion, PLK1 enhances the proliferation, migration and invasion of gastric cancer MGC-803 cells through affecting EMT.