Serum homocysteine concentration as a marker for advanced diabetic nephropathy in a cohort of elderly patients.

BACKGROUND: Hyperhomocysteinemia has been linked with chronic kidney disease (CKD). The present study investigated whether homocysteine (Hcy) serum levels might serve as a marker for the advancement of diabetic nephropathy (DN). METHODS: Clinical and laboratory indicators including Hcy, vitamin D (VD), urine protein, estimated glomerular filtration rate (eGFR) and the urinary protein/creatinine ratio in subjects > 65 years with DN (n = 1,845), prediabetes (n = 1,180) and in a non-diabetes (control) group (n = 28,720) were analyzed. RESULTS: DN patients had elevated Hcy concentrations, decreased VD and higher urinary protein levels, a reduced eGFR and a higher urinary protein/creatinine ratio compared with prediabetic and control subjects. After correcting for urinary protein quantitation, multivariate analysis revealed that both the Hcy concentration (P < 0.010) and urinary protein/creatinine ratio (P < 0.001) were risk factors, while the VD2 + VD3 serum concentration (P < 0.001) was a protective factor for DN. Moreover, Hcy > 12 µmol/L was a cut-off value for predicting advanced DN. CONCLUSION: Hcy serum concentration is a potential marker for the advancement of CKD in DN but not prediabetes patients.

Case report of pyogenic meningitis after tooth extraction in an elderly diabetic patient.

INTRODUCTION: Among elderly diabetic patients with comorbid conditions, the clinical characteristics of suppurative meningitis may not be typical, which is easy to cause misdiagnosis and delayed treatment. CASE PRESENTATION: In this report, we described the case of a 65-year-old elderly diabetic male who developed purulent meningitis after tooth extraction. The patient developed repeated infections (fever, headache and other symptoms) and was diagnosed with pyogenic meningitis by combining clinical manifestations and cerebrospinal fluid testing. The patient was treated with intracranial pressure reduction, nutritional support and anti-infection drugs, and recovered and was discharged after 15 days. CONCLUSIONS: The atypical clinical presentation and insidious onset of the disease in this patient could easily lead to missed diagnosis and misdiagnosis. We emphasize that we should be vigilant with purulent meningitis and active treatment and prevention should be implemented in this subgroup of patients. During treatment, active anti-infection and nutritional support should be provided on the basis of blood glucose control.

Clinical progression and outcomes of 260 patients with severe COVID-19: an observational study.

This paper presents the results of an observational, prospective study of the clinical progression and outcomes of patients with severe COVID-19. Overall, 260 patients with severe COVID-19 were included. The median age of the patients was 61 years (IQR 42.0-73.0), and 119 (45.8%) patients had one or more medical comorbidities. The median time from initial onset of symptoms to hospital admission was 8 days (IQR 6.0-11.0). Varying degrees of abnormalities in blood biochemical results were detected in most patients. All patients received supportive therapy and antiviral treatment. All patients were administered empirical antibiotic treatment with a median time of 5 days (IQR 3-7). Mechanical ventilation was required in accordance with respiratory conditions. At the data cutoff, 183 (70.4%) patients had been discharged, and 17 (6.5%) patients had been transferred to the intensive care unit (ICU). Twenty-five (9.6%) patients had died, and 35 (13.5%) patients were still in the hospital. During follow-up, 7 patients with fever were negative for SARS-Cov-2 antigens upon retest. The implications of the results are discussed for clinical features and the management of patients with severe COVID-19.

Neurocognitive Outcomes in Adult Quasi -Moyamoya Disease: A Prospective Analysis of Consecutive Cases.

BACKGROUND: This study aimed to evaluate and compare the neurocognitive outcomes of adult quasi-moyamoya disease (quasi-MMD) patients with autoimmune diseases (AIDs) to help better manage these patients. METHODS: We performed a structured battery of neurocognitive tests to analyze and compare the neurocognitive outcomes of adult quasi-MMD patients with AID in our hospital from October 2000 to September 2015. RESULTS: Overall, 27.3% of the neuropsychological test comparisons indicated a significant improvement in cognition, and a significant decline was found in 6%. In 47.4% of comparisons, the observed difference did significantly change the reliable change indices (RCI) before and after anti-autoimmune treatment. We found that the number of patients showing significant improvements, and no change in cognitive outcomes did differ between quasi-MMD and MMD (31.8% vs 14.9% with p = 0.006 and 50.0% vs 66.8% with p = 0.031, respectively; Chi-squared test). The incidence of cognitive decline in quasi-MMD patients (18.2%) did not significantly differ from that in MMD patients (18.3%) (p = 0.982). After adjusting for covariates, including sex, age, type 2 diabetes mellitus, risk factors, other comorbidities, and AID, multiple logistic regression analysis suggested that AID was more likely to aggravate the neurocognitive outcome of quasi-MMD patients (p = 0.042, odds ratio (OR) 6.78, 95% confidence interval (CI) 1.31-62.71). CONCLUSIONS: AID was more likely to aggravate the neurocognitive outcome of quasi-MMD patients, and anti-autoimmune treatment could improve long-term neurocognitive outcomes. These findings indicated that AID seemed to be an independent risk factor for the pathological and physiological mechanisms of quasi-MMD.

Relationship of gender and age on thyroid hormone parameters in a large Chinese population

ABSTRACT Objective This study aimed to present the impact of age and gender on thyroid hormone levels in a large Chinese population with sufficient iodine intake. Subjects and methods A total of 83643 individuals were included and were stratified by age and gender. The median, 2.5th and 97.5th of thyrotropin (TSH), free triiodothyronine (FT3), free thyroxine (FT4) and FT3/FT4 ratio were calculated for both genders for every decade from 18 to over 80 years. TSH, FT3, FT4, FT3/FT4 distribution in each age group was evaluated for females and males using smoothing splines in the generalized additive models (GAM). TSH concentrations were compared in the different age groups in gender. Results In the over 80s age group, the TSH level (median: 2.57 mIU/L, 2.5th-97.5th: 0.86-7.56 mIU/L) was significantly higher than other age groups, irrespective to gender (P<0.001). Females had a higher TSH value than males in all age groups (P<0.001). Results of the smoothing curves showed that TSH increased with age, FT3 concentration was higher in males than in females and the tendency of the FT3/FT4 ratio was basically similar to that of FT3. TSH concentration in the 50s age group (median 2.48 mIU/L for females versus 2.00 mIU/L for males) was significantly higher than that in the 30s age group (median 2.18 mIU/L for females versus median 1.85 mIU/L for males). Conclusions In accord with increasing TSH values during aging, females and older adults have lower FT3 values and lower FT3/FT4 ratios, while the FT4 values remain stable.

Relationship of gender and age on thyroid hormone parameters in a large Chinese population.

Objective This study aimed to present the impact of age and gender on thyroid hormone levels in a large Chinese population with sufficient iodine intake. Subjects and methods A total of 83643 individuals were included and were stratified by age and gender. The median, 2.5th and 97.5th of thyrotropin (TSH), free triiodothyronine (FT3), free thyroxine (FT4) and FT3/FT4 ratio were calculated for both genders for every decade from 18 to over 80 years. TSH, FT3, FT4, FT3/FT4 distribution in each age group was evaluated for females and males using smoothing splines in the generalized additive models (GAM). TSH concentrations were compared in the different age groups in gender. Results In the over 80s age group, the TSH level (median: 2.57 mIU/L, 2.5th-97.5th: 0.86-7.56 mIU/L) was significantly higher than other age groups, irrespective to gender (P<0.001). Females had a higher TSH value than males in all age groups (P<0.001). Results of the smoothing curves showed that TSH increased with age, FT3 concentration was higher in males than in females and the tendency of the FT3/FT4 ratio was basically similar to that of FT3. TSH concentration in the 50s age group (median 2.48 mIU/L for females versus 2.00 mIU/L for males) was significantly higher than that in the 30s age group (median 2.18 mIU/L for females versus median 1.85 mIU/L for males). Conclusions In accord with increasing TSH values during aging, females and older adults have lower FT3 values and lower FT3/FT4 ratios, while the FT4 values remain stable.

[Mutation analysis of a family affected with isolated proteinuria].

OBJECTIVE: To analyze the clinical characteristics and genetic features of a family affected with isolated proteinuria. METHODS: Clinical data of the family was collected. Mutations of 191 renal disease-related genes in the proband were screened with next generation sequencing (NGS). Sanger sequencing was used to verify suspected mutations in his family members and 100 healthy controls. The impact of the mutation was predicted with online software SIFT. Frequency of the mutation was searched in databases including 1000 Genomic Project, ESP and ExAC. RESULTS: NGS and Sanger sequencing showed that the proband harbored compound heterozygous mutations of ADCK4 gene including c.748C>G (p.Asp250His) and c.1041G>T (p.Cys347*), which were respectively inherited from his mother and father whom were both non-symptomatic. CONCLUSION: The proband may have ADCK4-associated glomerulopathy due to the compound heterozygous mutations of the ADCK4 gene.

PIWIL1 suppresses circadian rhythms through GSK3ß-induced phosphorylation and degradation of CLOCK and BMAL1 in cancer cells.

Circadian rhythms are maintained by series of circadian clock proteins, and post-translation modifications of clock proteins significantly contribute to regulating circadian clock. However, the underlying upstream mechanism of circadian genes that are responsible for circadian rhythms in cancer cells remains unknown. PIWIL1 participates in many physiological processes and current discoveries have shown that PIWIL1 is involved in tumorigenesis in various cancers. Here we report that PIWIL1 can suppress circadian rhythms in cancer cells. Mechanistically, by promoting SRC interacting with PI3K, PIWIL1 can activate PI3K-AKT signalling pathway to phosphorylate and inactivate GSK3ß, repressing GSK3ß-induced phosphorylation and ubiquitination of CLOCK and BMAL1. Simultaneously, together with CLOCK/BMAL1 complex, PIWIL1 can bind with E-BOX region to suppress transcriptional activities of clock-controlled genes promoters. Collectively, our findings first demonstrate that PIWIL1 negatively regulates circadian rhythms via two pathways, providing molecular connection between dysfunction of circadian rhythms and tumorigenesis.

RAE1 promotes BMAL1 shuttling and regulates degradation and activity of CLOCK: BMAL1 heterodimer.

Circadian rhythm is an autoregulatory rhythm, which is sustained by various mechanisms. The nucleocytoplasmic shuttling of BMAL1 is essential for CLOCK translocation between cytoplasm and nucleus and maintenance of the correct pace of the circadian clock. Here we showed that RAE1 and NUP98 can promote the degradation of BMAL1 and CLOCK. Knockdown of RAE1 and NUP98 suppressed BMAL1 shuttling, leading to cytoplasm accumulation of CLOCK. Furthermore, Chip assay showed that knockdown of RAE1 and NUP98 can enhance the interaction between CLOCK: BMAL1 and E-box region in the promoters of Per2 and Cry1 while reducing its transcription activation activity. Our present study firstly revealed that RAE1 and NUP98 are critical regulators for BMAL1 shuttling.

Vitamin D status is associated with 1,5-anhydro-d-glucitol status in patients with type 2 diabetes mellitus.

Serum 1,5-anhydro-d-glucitol (1,5-AG) concentrations are short-term hyperglycemia indicators and were used to estimate the effects of serum vitamin D concentrations on glycemic control in patients with type 2 diabetes mellitus (T2DM). Serum concentrations of 1,5-AG, 25-hydroxyvitamin D2 (25-OH-D2), and 25-hydroxyvitamin D3 (25-OH-D3) from 11 026 patients with T2DM, hospitalized in the Department of Endocrinology, Shanghai Central Hospital of Xuhui District, from January 2012 to June 2015, were retrospectively analyzed. Correlation analyses revealed correlations between 1,5-AG and 25-OH-D3 (r = 0.05, P < 0.001), age (r = 0.05, P < 0.001), and 25-OH-D2 + 25-OH-D3 (25-OH-D2/D3) (r = 0.05, P < 0.001). Linear regression analyses revealed associations between 1,5-AG and 25-OH-D2/D3 (adjusted R2 = 0.003) as well as 25-OH-D3 (adjusted R2 = 0.002). In males with 1,5-AG levels ≤11.55 mg/L, serum concentrations of 25-OH-D2 (P < 0.001) and 25-OH-D3 (P = 0.001) were significantly lower than those in diabetic males with 1,5-AG levels >11.55 mg/L. Serum concentrations of 25-OH-D2/D3 in patients with T2DM were associated with 1,5-AG retention, suggesting involvement in glycemic control.

PIWIL2 suppresses Siah2-mediated degradation of HDAC3 and facilitates CK2α-mediated HDAC3 phosphorylation.

HDAC3 is involved in deacetylation of histone and non-histone proteins, having a key role in the regulation of gene transcription and also in the process of tumorgenesis. However, how HDAC3 is regulated in cancer remains largely unclear. Here, we showed that PIWIL2 can interact with HDAC3, leading to stabilization of HDAC3 from ubiquitin-mediated degradation by competitive association with E3 ubiquitin ligase Siah2. Furthermore, we found that expression of PIWIL2 enhanced HDAC3 activity via CK2α. PIWIL2 facilitated the interaction between HDAC3 and CK2α, thus exhibiting a promotion on the HDAC3 phosphorylation by CK2α. Further work showed that PIWIL2 could promote cell proliferation and suppress cell apoptosis via regulating HDAC3. Our present study firstly revealed that PIWIL2 can play a role in HDAC3-mediated epigenetic regulation on cancer cell proliferation and apoptosis. These findings provide a novel insight into the roles of PIWIL2 in tumorigenesis.

Overexpression of secretagogin promotes cell apoptosis and inhibits migration and invasion of human SW480 human colorectal cancer cells.

OBJECTIVE: In order to investigate the effect of secretagogin (SCGN) on colorectal cancer (CRC) cells apoptosis, invasion and migration in vitro. METHODS: Expression of SCGN in CRC tissues and the paired adjacent non-tumorous tissues (n = 36) and four human CRC cell lines (HT29, HCT116, SW480 and SW620) were detected. SW480 cells were transfected with the SCGN overexpression plasmid (eGFP-SCGN), si-SCGN-773, and the corresponding negative controls (NCs). Then, cell-cycle distribution, cell apoptosis, migration, invasion and expression of apoptosis- and metastasis-related proteins were detected. RESULTS: SCGN was significantly downregulated in CRC tissues as compared with the adjacent non-tumorous tissues. The expression of SCGN in HT29 and SW480 cells were lower than those in HT116 and SW620 cells. We transfected SW480 cells with SCGN overexpression plasmid eGFP-SCGN and found the increased cell apoptosis, with cell arresting at G0/G1 phase. SW480 cells with SCGN overexpression showed wider wound width and fewer invaded cells than control and blank cells, with upregulated Bax, cleaved Caspase 3 and E-cadherin, and downregulated Bcl-2 and Vimentin. We also transfected SW480 cells with si-SCGN-773 and found si-SCGN increased cell migration and invasion, but did not affect cell apoptosis and expression of related proteins. CONCLUSION: We concluded that the overexpression of SCGN in SW480 cells promoted cell apoptosis and inhibited cell migration and invasion.

Itraconazole attenuates hepatic gluconeogenesis and promotes glucose uptake by regulating AMPK pathway.

The primarily metabolic abnormality in type 2 diabetes mellitus (T2DM) is the defect in gluconeogenesis and glucose uptake. Itraconazole (ITCZ) is a traditional azole drug with anti-fungal and anticancer properties. However, limited attention has been directed towards the contribution of ITCZ to hepatic gluconeogenesis and glucose uptake in T2DM. The present study aimed to investigate the potential effects of ITCZ on hepatic gluconeogenesis and glucose uptake as well as the underlying mechanisms. No obvious change in cell viability was detected by MTT assay in HepG2 cells with ITCZ treatment at gradually increasing concentrations. Western blot analysis demonstrated that the phosphorylation level of 5' adenosine monophosphate-activated protein kinase (AMPK) was significantly elevated by ITCZ treatment at ≥5 µg/ml (P<0.05). Moreover, ITCZ repressed the gluconeogenesis of HepG2 cells, as evidenced by the dose-dependently increased glycogen synthase kinase 3ß phosphorylation level and a notably decreased glucose production rate (P<0.05). Simultaneously, the expression of peroxisome proliferator-activated receptor γ co-activator 1α, phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) in HepG2 cells was reduced by ITCZ in a dose-dependent manner (P<0.001). Furthermore, a 2-deoxyglucose uptake assay revealed that the glucose uptake of HepG2 cells was notably enhanced, accompanied by the ITCZ dose-dependent upregulation of glucose transporter-4 (GLUT-4) (P<0.05). Conversely, silencing of AMPK by small interfering RNA resulted in an increase of ITCZ-reduced gluconeogenesis and inhibition of ITCZ-induced glucose uptake with relative upregulation of PEPCK and G6Pase and downregulation of GLUT4 in the presence of 50 µg/ml ITCZ (P<0.05). Overall, the results indicated that AMPK has an important role in regulating ITCZ-induced glucose uptake by stimulating GLUT4 in HepG2 cells. Therefore, ITCZ may become a promising candidate for T2DM therapy.

CLOCK interacts with RANBP9 and is involved in alternative splicing in spermatogenesis.

The core circadian gene CLOCK plays an important role in regulating male reproduction. However, the underlying mechanism still remains unclear. In the present study, we executed yeast two-hybrid screening using cDNA fragment of CLOCK PAS A domain as bait, and identified RANBP9 as a novel protein interacting with CLOCK. The interaction between CLOCK and RANBP9 was further validated by in vivo and in vitro assays. Previous studies have confirmed that SF3B3 was a RANBP9 interacting protein. Subsequently, our study also found that CLOCK and SF3B3 can interact with each other by co-immunoprecipitation in mouse testis. In order to dissect the underlying mechanism of CLOCK in spermatogenesis, we also performed RNA-immunoprecipitation followed by high-throughput sequencing (RIP-Seq) in mouse testis. The result of sequence analyses and Gene Ontology enrichment analyses (biological processes) demonstrated that CLOCK can directly bind 186 key mRNA transcripts in mouse spermatogenesis. Taken together, our results firstly showed that CLOCK can interact with RANBP9 and bind with mRNAs, demonstrating that CLOCK is involved in alternative splicing in spermatogenesis. These results reveal a novel mechanism for CLOCK in spermatogenesis.

Cancer/testis antigen PIWIL2 suppresses circadian rhythms by regulating the stability and activity of BMAL1 and CLOCK.

Circadian rhythms are regulated by transcriptional and post-translational feedback loops generated by appropriate functions of clock proteins. Rhythmic degradation of the circadian clock proteins is critical for maintenance of the circadian oscillations. Notably, circadian clock does not work during spermatogenesis and can be disrupted in tumors. However, the underlying mechanism that suppresses circadian rhythms in germ cells and cancer cells remains largely unknown. Here we report that the cancer/testis antigen PIWIL2 can repress circadian rhythms both in the testis and cancer cells. By facilitating SRC binding with PI3K, PIWIL2 activates the PI3K-AKT pathway to phosphorylate and deactivate GSK3ß, suppressing GSK3ß-induced phosphorylation and degradation of circadian protein BMAL1 and CLOCK. Meanwhile, PIWIL2 can bind with E-Box sequences associated with the BMAL1/CLOCK complex to negatively regulate the transcriptional activation activity of promoters of clock-controlled genes. Taken together, our results first described a function for the germline-specific protein PIWIL2 in regulation of the circadian clock, providing a molecular link between spermatogenesis as well as tumorigenesis to the dysfunction of circadian rhythms.

Clock1a affects mesoderm development and primitive hematopoiesis by regulating Nodal-Smad3 signaling in the zebrafish embryo.

Circadian clock and Smad2/3/4-mediated Nodal signaling regulate multiple physiological and pathological processes. However, it remains unknown whether Clock directly cross-talks with Nodal signaling and how this would regulate embryonic development. Here we show that Clock1a coordinated mesoderm development and primitive hematopoiesis in zebrafish embryos by directly up-regulating Nodal-Smad3 signaling. We found that Clock1a is expressed both maternally and zygotically throughout early zebrafish development. We also noted that Clock1a alterations produce embryonic defects with shortened body length, lack of the ventral tail fin, or partial defect of the eyes. Clock1a regulates the expression of the mesodermal markers ntl, gsc, and eve1 and of the hematopoietic markers scl, lmo2, and fli1a Biochemical analyses revealed that Clock1a stimulates Nodal signaling by increasing expression of Smad2/3/4. Mechanistically, Clock1a activates the smad3a promoter via its E-box1 element (CAGATG). Taken together, these findings provide mechanistic insight into the role of Clock1a in the regulation of mesoderm development and primitive hematopoiesis via modulation of Nodal-Smad3 signaling and indicate that Smad3a is directly controlled by the circadian clock in zebrafish.

PIWIL1 destabilizes microtubule by suppressing phosphorylation at Ser16 and RLIM-mediated degradation of Stathmin1.

Human PIWIL1, alias HIWI, is a member of Piwi protein family and expressed in various tumors. However, the underlying mechanism of PIWIL1 in tumorigenesis remains largely unknown. Stathmin1 is a cytosolic phosphoprotein which has a critical role in regulating microtubule dynamics and is overexpressed in many cancers. Here we report that PIWIL1 can directly bind to Stathmin1. Meanwhile, PIWIL1 can up-regulate the expression of Stathmin1 through inhibiting ubiquitin-mediated degradation induced by an E3 ubiquitin ligase RLIM. Furthermore, PIWIL1 can also reduce phosphorylation level of Stathmin1 at Ser-16 through inhibiting the interaction between CaMKII and Stathmin1. Our results showed that PIWIL1 suppresses microtubule polymerization, and promotes cell proliferation and migration via Stathmin1 for the first time. Our study reveals a novel mechanism for PIWIL1 in tumorigenesis.

Dipeptidyl peptidase 4 concentration influenced by serum insulin levels rather than arterial stiffness index in type 2 diabetics.

OBJECTIVE: To investigate whether inhibitors of dipeptidyl peptidase-4 (DPP-4) can suppress the atherosclerotic development in diabetic patients. METHODS: The prospective study was carried out in the Out-patient Department of XuHui Central Hospital, Shanghai, China, between March and August 2013. The correlation of major index of glucose and lipid metabolism profiles, and the arterial stiffness index (AI) between diabetic patients and healthy subjects were analyzed. PATIENTS OR MATERIALS: 39 patients with type 2 diabetes and 29 healthy subjects were enrolled for measurements of blood glucose, plasma insulin, HbA1c, TC, cholesterol, HDL, AI and body mass index (BMI). RESULTS: Significant differences were found between DPP-4 and blood glucose (fasting and 2 h postprandial), HbA1c, cholesterol, high density lipoprotein and arterial stiffness in normal subjects and diabetic patients. Only the fasting insulin concentration and high density lipoprotein had a significant impact on DPP-4 levels. CONCLUSIONS: It was clear that insulin (fasting) and HDL levels had an impact on DPP-4 activity but only in patients with Type 2 diabetes. The arterial stiffness index was not correlated with DPP-4 levels in Type 2 diabetic patients.

PIWIL2 induces c-Myc expression by interacting with NME2 and regulates c-Myc-mediated tumor cell proliferation.

c-Myc serves as a crucial regulator in multiple cellular events. Cumulative evidences demonstrate that anomalous c-Myc overexpression correlates with proliferation, invasion and metastasis in various human tumors. However, the transcriptionally activating mechanisms responsible for c-Myc overexpression are complex and continue to be intangible. Here we showed that Piwi-Like RNA-Mediated Gene Silencing 2 (PIWIL2) can upregulate c-Myc via binding with NME/NM23 nucleoside diphosphate kinase 2 (NME2). PIWIL2 promotes c-Myc transcription by interacting with and facilitating NME2 to bind to G4-motif region within c-Myc promoter. Interestingly, in a c-Myc-mediated manner, PIWIL2 upregulates RhoA, which in turn induces filamentary F-actin. Deficiency of PIWIL2 results in obstacle for c-Myc expression, cell cycle progress and cell proliferation. Taken together, our present work demonstrates that PIWIL2 modulates tumor cell proliferation and F-actin filaments via promoting c-Myc expression.