Effect of 1,25(OH2D3 on the proliferation of human mesangial cells and their expression of Ki67.

Previous studies have found that 1,25-dihydroxyvitamin D3 [1,25(OH)2D3 or VD3] exerts many biological effects, including the inhibition of cell proliferation and induction of apoptosis, but its mechanism of action remains unclear. The goal of our investigation was to explore the effects of 1,25(OH)2D3 on the proliferation of cultured human mesangial cells and their expression of Ki67 in vitro, and to establish its mechanism of action. Cultured human mesangial cells were randomly divided into the following four groups: normal control (N group; administered Dulbecco's modified Eagle's medium containing 5% fetal bovine serum), proliferation [epidermal growth factor (EGF) group; administered 10 µg/L EGF], VD3 intervention [administered 10-8 M 1,25(OH)2D3], and proliferation and intervention [EGF+VD3 group; administered 10 µg/L EGF and 10-8 M 1,25(OH)2D3]. Cells were incubated for 48 h with the corresponding treatment, and fluorescence immunocytochemistry and reverse transcription-quantitative polymerase chain reaction were used to detect expression of Ki67 protein and mRNA, respectively. Compared to the N group, Ki67 levels were found to be higher in the EGF group but significantly lower in the VD3 intervention group. Moreover, expression of Ki67 by cells in the EGF+VD3 group was significantly lower than that of those in the EGF group. All of these differences were statistically significant (P < 0.05). In conclusion, 1,25(OH)2D3 inhibited Ki67 expression and the proliferation of human mesangial cells; therefore, Ki67 may be regarded as a potent therapeutic target in mesangial proliferative glomerulonephritis.

Synonymous codon selection in the hepatitis B virus translation initiation region.

Hepatitis B virus (HBV) infection is a major health problem worldwide. This virus and its hosts are often fated to continual co-evolutionary interactions. Codon usage analysis has significance for studies of co-evolution between viruses, their hosts, and mRNA translation. Adaptation of the overall codon usage pattern of HBV to that of humans is estimated using the synonymous codon usage value (RSCU), and the synonymous codon usage biases for the translation initiation region (TIR) of HBV are analyzed by calculation of the usage fluctuation of each synonymous codon along the TIR (the first 50 codon sites of the whole coding sequence of HBV). With respect to synonymous codon usage, our results demonstrated that HBV had no significant tendency to select over-represented codons, but had a significant tendency to select certain under-represented codons in the viral genome. Within the three common HBV hosts, 14 of 59 codons had a similar usage pattern, suggesting that mutation pressure from this DNA virus played an important role in the formation of virus synonymous codon usage. In addition, there was no obvious trend for the codons with relatively low energy to be highly selected in the TIR of HBV, suggesting that the synonymous codon usage patterns for the TIR might not be affected by the nucleotide sequence secondary structure; however, synonymous codon usage in the TIR of HBV was influenced by the overall codon usage patterns of the hosts to some degree. Our results suggest that mutation pressure from HBV plays an important role in the formation of synonymous codon usage of the viral genome, while translation selection from the hosts contributes to virus translational fine-tuning.

Ultrasonic bone curette in thoracic spinal decompression: a comprehensive systematic review and meta-analysis.

OBJECTIVE: The aim of this study was to compare the efficacy and safety of ultrasonic bone curette (UBC) and conventional surgical instruments in thoracic laminectomy decompression (TLD) for the treatment of thoracic spinal stenosis (TSS) by meta-analysis. MATERIALS AND METHODS: Two authors independently searched Medline via PubMed, Embase, Cochrane Library, Web of Science, Wanfang Database, and China National Knowledge Infrastructure for the period from the establishment of the database until January 2023 to identify the studies on the safety and efficacy of UBC vs. conventional instruments for TSS. Data extraction and quality assessment were performed by two researchers independently. We used RevMan 5.4 software (Review Manager Web, The Cochrane Collaboration, Copenhagen, Denmark) to analyze the data. RESULTS: Eight retrospective studies were included in the present work. This meta-analysis revealed that no significant differences in the preoperative JOA scores, the JOA scores at the last follow-up, the improvement rate of JOA scores, and the incidence of cerebrospinal fluid leakage/dura injury were detected between the two groups (p>0.05). However, there were significant differences in the operative time and intraoperative blood loss during single-level TLD [operative time: MD=-1.47, 95% CI (-1.86, -1.09), p<0.001; intraoperative blood loss: MD=-46.62, 95% CI (-53.83, -39.40), p<0.001], total operative time [MD=-56.88, 95% CI (-69.66, -44.10), p<0.001], total intraoperative blood loss [MD=-143.52, 95% CI (-212.49, -74.54), p<0.001], the incidence of neurological deterioration/nerve root injury [RR= 0.29, 95% CI (0.09, 0.91), p=0.03] between the groups. CONCLUSIONS: The application of UBC in TLD to treat TSS is safe and effective. UBC can significantly shorten operation time and reduce intraoperative blood loss compared to traditional surgical instruments. Moreover, it has the advantage of reducing perioperative nerve injury.

cAMP/PKA/CREB signaling pathway-mediated effects of melatonin receptor genes on clock gene expression in Bactrian camel ovarian granulosa cells.

The cAMP/PKA/CREB pathway is involved in the regulation of melatonin during important physiological activities in mammals. However, the regulation of circadian clock genes in ovarian granulosa cells remains unclear. Herein, we determined the relationship between melatonin and biological clock genes using cultured Bactrian camel ovarian granulosa cells. The enzyme-linked immunosorbent assays showed that the cAMP content was reduced when melatonin receptor (MT) genes or cryptochrome (Cry) genes were overexpressed; the quantitative polymerase chain reaction and western blot analyses revealed that the expression levels of all circadian clock genes (GNB2, PKA, CREB, Per1/2/3, and Clock) except Cry1/2 decreased significantly at 24 h. Cellular immunolocalization analysis showed that melatonin receptors were localized in the cell membrane and cytoplasm; the CRY protein was mainly localized in the nucleus. Overall, our findings indicated that the rhythmic regulation of ovarian granulosa cells was consistent with the regulatory action of the central circadian clock.

Association study of SNCA gene polymorphisms with schizophrenia in a Chinese North Han population.

OBJECTIVE: Previous studies suggested that the alpha-synapse protein (SNCA) gene and its coding product α-synuclein (α-Syn) may play a role in the pathogenesis of neurodegenerative diseases. The mutation of SNCA can influence the formation of nerve fibers and the function of dopaminergic neurons, and that may be related to addictive behavior, such as alcohol dependence. SNCA may overlap with the pathogenesis of schizophrenia and Parkinson's disease or alcohol dependence associated with the dopamine pathway. The aim was to determine the association between three SNCA SNPs (rs3822086C/T, rs11931074G/T, and rs356219A/G) and schizophrenia in a Chinese North Han population. PATIENTS AND METHODS: A total of 878 subjects, with or without schizophrenia, were included in our study. DNA purification, Polymerase Chain Reaction (PCR) amplification, and subsequent restriction fragment length polymorphism (RFLP) analysis were manipulated to determine genotypes. RESULTS: Between the schizophrenia group and healthy group, neither the genotype nor allele frequencies of rs3822086C/T, rs11931074G/T, or rs356219A/G differed significantly in either the total sample or the subgroups. In the haplotype analysis, the ATT and GTT haplotype frequencies differed significantly between the patients and controls in the total sample (χ2=6.052, p=0.0139; χ2=4.508, p=0.0337). In the female subgroup, the ATT haplotype frequency differed significantly between the patients and controls (χ2=4.219, p=0.04). CONCLUSIONS: There was no association between SNCA polymorphisms and schizophrenia in the North Han Chinese population, and the ATT haplotype may be a susceptibility factor for schizophrenia.

EHMT2 promotes the development of prostate cancer by inhibiting PI3K/AKT/mTOR pathway.

OBJECTIVE: To investigate the expression characteristics and the potential role of euchromatic histone-lysine N-methyltransferase 2 (EHMT2) in the clinical pathology and prognosis of prostate cancer (PCa). PATIENTS AND METHODS: Quantitative Real-time polymerase chain reaction (qRT-PCR) was performed to detect the expression of EHMT2 in 55 pairs of PCa tissues and adjacent normal tissues. The relationship between EHMT2 expression and the pathological features, as well as the prognosis of PCa patients was analyzed. EHMT2 expression in PCa cells was determined by qRT-PCR as well. In addition, EHMT2 knockdown model was constructed by transfection of the small interfering RNA in PCa cell lines PC-3 and DU-145. The regulatory effects of EHMT2 on the behaviors of PCa cells were evaluated through cell counting kit-8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (EdU), colony formation assay and flow cytometry. Finally, we detected the protein levels of relative genes in PI3K/AKT/mTOR pathway through Western blot. RESULTS: QRT-PCR results showed that the expression level of EHMT2 in PCa tissues was markedly higher than that in adjacent normal tissues. Compared with PCa patients with low expression of EHMT2, those with high expression of EHMT2 had higher pathological grade and lower overall survival. EHMT2 was also highly expressed in PCa cell lines. Knockdown of EHMT2 inhibited the proliferative potential and induced apoptosis of PCa cells. Western blot results revealed that PCa cells with EHMT2 knockdown presented downregulated p-PI3K, p-AKT and p-mTOR. CONCLUSIONS: EHMT2 was highly expressed in PCa, and its expression is closely correlated with tumor stage and poor prognosis of PCa patients. EHMT2 promoted the malignant progression of PCa by inhibiting PI3K/AKT/mTOR pathway.

Peripheral Blood Leukocyte Telomere Length Is Associated with Age but Not Renal Function: A Cross-Sectional Follow-Up Study.

OBJECTIVES: We aimed to evaluate the relationship between baseline renal function and changes in telomere length in Han Chinese. METHODS: The telomere restriction fragment (TRF) length of leukocytes in the peripheral blood was measured in healthy volunteers recruited in 2014. The estimated glomerular filtration rate (eGFR) was calculated based on serum creatinine (Scr) and serum cystatin C (CysC)-eGFRcys and eGFRScr-cys through the Cockcroft-Gault formula (eGFRC-G) or the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI / eGFRCKD-EPI) equation. The correlation between telomere length changes over time and renal function was analyzed. RESULTS: Leukocyte TRF lengths were negatively correlated to age (r = -0.393, p < 0.001) and serum CysC (r = -0.180, p < 0.01), while positively associated with eGFRCKD-EPI, eGFRC-G, eGFRcys, and eGFRScr-cys (r = 0.182, 0.122, 0.290, and 0.254 respectively, p < 0.01). The 3-year change of telomere length was 46 bp/years. When adjusted for age, the associations between telomere length changes and baseline, subsequent TRF lengths, and serum CysC were no longer present. No association was observed between TRF length changes and renal function. CONCLUSION: The rate of telomere length changes was affected by age and baseline telomere length. The telomere length changes might be important markers for aging.

Identification of potential therapeutic target genes and mechanisms in non-small-cell lung carcinoma in non-smoking women based on bioinformatics analysis.

OBJECTIVE: The study was aimed to explore the underlying mechanisms and identify the potential target genes by bioinformatics analysis for non-small-cell lung carcinoma (NSCLC) treatment in non-smoking women. MATERIALS AND METHODS: The microarray data of GSE19804 was downloaded from Gene Expression Omnibus (GEO) database. Paired samples (from the same patient) of tumor and normal lung tissues from 60 non-smoking female NSCLC patients were used to identify differentially expressed genes (DEGs). The functional enrichment analysis was performed. Furthermore, the protein-protein interaction (PPI) network of the DEGs was constructed by Cytoscape software. The module analysis was performed. RESULTS: Totally, 817 DEGs including 273 up- and 544 down-regulated genes were identified. The up-regulated genes were mainly enriched in extracellular matrix (ECM)-receptor interaction, focal adhesion and cell cycle functions, while down-regulated genes were mainly enriched in the cytokine-cytokine receptor interaction pathway. DEGs including hyaluronan-mediated motility receptor (HMMR), collagen, type I alpha 2 (COL1A2), cyclin A2 (CCNA2), MAD2 mitotic arrest deficient-like 1 (MAD2L1), interleukin 6 (IL6) and interleukin 1, beta (IL1B) were identified in these functions. These genes were hub nodes in PPI networks. Besides, there were 3 up-regulated modules and 1 down-regulated module. The significant pathways were ECM-receptor interaction and focal adhesion in up-regulated modules, while in down-regulated module, the significant pathway was mitogen-activated protein kinase (MAPK) signaling pathway. CONCLUSIONS: The ECM-receptor interaction, focal adhesion, cell cycle and cytokine-cytokine receptor interaction functions may be associated with NSCLC development. Genes such as HMMR, COL1A2, CCNA2, MAD2L1, IL6 and IL1B may be potential therapeutic target genes for NSCLC.

Human DNA Polymerase-beta Promoter: Phorbol Ester Activation Is Mediated through the cAMP Response Element and cAMP-Response-Element-Binding Protein.

That mammalian DNA polymerase-beta (beta-pol) gene transcription is upregulated by activated ras and also by phorbol ester (TPA) treatment suggests the involvement of protein kinase C in the gene expression control for this DNA repair enzyme. Yet, the core promoters of the human, bovine and rodent beta-pol genes do not have a TPA response element or other binding site for the transcriptional activator AP-1. Instead, these beta-pol promoters appear to be regulated mainly by proteins binding to the cAMP response element (CRE) centered within 50 bp 5' of the transcriptional start site. In this study, the CRE in the human beta-pol promoter was found to mediate TPA upregulation of the cloned promoter in HeLa cell transient expression experiments. To further examine the role of this CRE in TPA stimulation, we used several mutated promoters that were either deficient in protein binding to the CRE or contained extra CRE sites arranged as tandem repeats. All constructs with at least one functional CRE were upregulated by TPA, whereas mutants lacking CRE protein-binding function were not TPA upregulated. Analyses of HeLa nuclear extract DNA-binding proteins indicated that the beta-pol CRE was bound by CRE-binding protein (CREB) family members CREB-1 and activating transcription factor-1, but not by AP-1 or complexes containg AP-1 subunits. These results suggest that CREB, rather than AP-1 proteins, are required for the CRE-mediated TPA activation of the beta-pol promoter.

Pharmacological analysis of functional neurovascular transmission in canine splenic arteries: role of neuropeptide Y.

1 The effects of neuropeptide Y (NPY) upon the isolated vasculature are reviewed. 2 The vasconstrictor responses to periarterial nerve stimulation (PNS) and neurotransmission by noradrenaline (NA) and ATP are discussed and illustrated using canine isolated perfused splenic artery. 3 Modulation of the vascular responses to PNS by NPY via pre- and post-junctional NPY Y2 and Y1 receptors is discussed. 4 Evidence is presented for different alpha1-adrenoceptor subtypes mediating vasoconstriction to exogenous and endogenously released NA and their different locations in the neurovascular junction and extrajunctional regions. 5 Activation of NPY Y1-receptors potentiates sympathetic nerve activated alpha1-adrenoceptor vasoconstriction. The proposal that the postjunctional alpha1B adrenoceptor may be linked to the NPY Y1-receptor and is responsible for co-operation between sympathetic and NPYergic interactions in the vasculature is discussed.

Mechanisms of release of ATP from vascular purinergic nerves.

1. The vasoconstrictor response to periarterial nerve electrical stimulation (PNS) and neurotransmission by ATP are discussed and illustrated, using canine isolated and perfused splenic arterial preparations. 2. The conditions for appearance of dominant purinergic constrictor response to PNS are discussed. 3. Modulation of the purinergic vasoconstrictor responses to PNS by several kinds of presynaptic receptor agonists and antagonists is reviewed. 4. Influences of purinergic responses to PNS by guanethidine, reserpine, tetrodotoxin (TTX) or omega-conotoxin GVIA (omegaCTX) are also reviewed. 5. Effects of imipramine and removal of the endothelium are discussed. 6. Evidence is presented for selective inhibition of purinergic responses to PNS by an adequate cold storage of the vessel. 7. The roles of ATP released by PNS in isolated canine splenic arteries are proposed.

Prejunctional AT(1) receptor subtype-dependent modification of neurotransmitter releases in canine isolated splenic arteries.

1. The regulation by angiotensin II (Ang II) formed locally on nerve-stimulated purinergic and adrenergic components of double-peaked vasoconstrictions in the canine splenic artery and Ang II receptor subtypes involved were investigated. 2. The perfusion of the precursor angiotensin I (Ang I, 0.1-1 nm) did not affect the vasoconstrictor responses to noradrenaline (NA, 0.03-1 nmol) and adenosine 5'-triphosphate (ATP, 0.03-1 micromol). The second component vasoconstrictor response to nerve stimulation was dose dependently potentiated by Ang I (0.1-1 nm). The first peaked constriction was slightly, but insignificantly increased. The potentiating effects of Ang I were abolished by KRH-594 (10 nm), a selective AT(1) receptor antagonist, but not by PD 123319 (1-10 nm), an AT(2) receptor antagonist. KRH-594 (10 nm) or PD 123319 (10 nm) never affected the vasoconstrictions to either NA or ATP. 3. The treatment with KRH-594 (1-10 nm) produced a greater inhibition on the second peaked response than the first one, although both of them were dose dependently inhibited. PD 123319 (1-10 nm) did not affect the vasoconstrictor responses induced by nerve stimulation. 4. Inhibition of angiotensin-converting enzyme with 10 nm enalaprilat reduced the second peaked response, having no significant inhibition on the first peaked response. A higher dose of enalaprilat (100 nm) produced a greater inhibition of the second peak than the first one. It reduced the second peak by approximately 65%, while the first peak was decreased approximately 35%. After treatment with enalaprilat, Ang I (1 nm) failed to enhance the neuronal vascular response. Enalaprilat at doses used did not affect the vasoconstrictions to either NA or ATP. 5. The present results indicate that endogenously generated Ang II may produce a more marked potentiation of adrenergic transmission than purinergic transmission via activation of prejunctional AT(1) receptors.

Prognostic improvement of reexcision for positive resection margins in patients with advanced gastric cancer.

AIM: The aim of this study was to investigate the effect of reexcision for advanced gastric cancer (GC) with positive resection margins on prognosis and to identify the selection criteria for the reexcision of patients with positive margins. PATIENTS AND METHODS: This was a retrospective study of 122 patients with positive margins who underwent potentially curative resection for locally advanced GC. The clinicopathological factors and survival among 50 patients who were reexcised to a negative resection margin (NR group) and 72 patients who were left with a positive resection margin (PR group) were compared using univariate and multivariate analyses. RESULTS: Median survival in the PR group was 18 months compared with 23 months in the NR group (p = 0.019). In the ≤ pN2-category subset, the PR group had a significantly worse prognosis compared with the NR group (median survival of 25 months vs. 44 months; p = 0.021). This difference was not observed in the pN3-category subset. In the univariate analysis, variables including pTNM stage, pN-category, and positive resection margin had adverse effects on OS among the entire population of 122 patients. A positive margin was confirmed as an independent prognostic factor for OS in the multivariate analysis. CONCLUSIONS: The reexcision of a positive margin improves the prognosis of patients with advanced GC, especially in those paitents with ≤ pN2-category disease and in patients undergoing D2 lymphadenectomy. Obtaining routine frozen sections of samples from the resection margin should be mandatory in the treatment of all GC patients undergoing potentially curative surgery.

Glucose-sensitive holographic sensors.

Holographic sensors for monitoring glucose were fabricated from hydrogel films containing chemical ligands based on phenylboronic acid. The films were transformed into reflection holograms using a diffusion method coupled with exposure to laser light. The diffraction wavelength of the holograms was used to monitor the swelling of the hydrogel film in the presence of glucose. Fully reversible changes in diffraction wavelength were demonstrated, highlighting the potential for using these holograms as glucose sensors.