Suppression of ovarian cancer by low-intensity ultrasound through depletion of IL-6/STAT3 inflammatory pathway-maintained cancer stemness.

Ovarian carcinoma is the key cause of cancer death from gynecological malignancy of women. Chemotherapy-resistance, metastasis and relapse contribute to the high mortality in ovarian cancer patients. Cancer stem cells (CSCs) stand for the root of kinds of cancer types such as ovarian cancer, are the key driver of tumor initiation, cancer metastasis, and resistance to conventional chemotherapy as well as genomic targeted therapy. Thus, the approach to eliminate CSCs and uncovering the mechanism will have substantial impact on cancer therapy. However, targeting CSC remains unfeasible in clinical practice in ovarian cancer therapy. In this study, we first found that Low-intensity ultrasound (LIUS) was capable of reducing the CSC populations in the xenograft model with ovarian cancer, with blocking survival, anti-apoptosis, self-renewal, and downregulating the cancer stemness genes in ovarian CSCs. Moreover, LIUS ameliorated IL-6/STAT3 inflammatory pathway via inhibiting IL-6-induced STAT3 phosphorylation, DNA binding activity and, the expressions of its downstream effectors in ovarian CSCs while no explicit effect was found in the corresponding bulk cancer cells. Additional approaches in molecular studies showed that LIUS disrupts CSC features via inhibiting IL-6/STAT3 inflammatory pathway. Collectively, our data for the first time elucidate IL-6/STAT3 inflammatory loop as the key CSC or cancer stemness pathway in ovarian cancer by LIUS treatment, providing a novel and potential therapy and a promising target in ovarian cancer.

Predictive Value of Quantitative Uterine Fibroid Perfusion Parameters From Contrast-Enhanced Ultrasound for the Therapeutic Effect of High-Intensity Focused Ultrasound Ablation.

OBJECTIVES: To assess the predictive significance of quantitative perfusion parameters from contrast-enhanced ultrasound (CEUS) for the therapeutic response to high-intensity focused ultrasound (HIFU) ablation in patients with uterine fibroids. METHODS: A total of 263 patients with single uterine fibroids were treated with HIFU ablation under ultrasound guidance. The arrival time, peak time, enhancement time, enhancement intensity, and enhancement rate were evaluated with pretreatment CEUS. According to a nonperfused volume ratio evaluation by posttreatment magnetic resonance imaging, all patients were assigned to groups with volume ratios of 70% or higher and lower than 70%. Then the predictive performances of different parameters for ablation efficacy were studied. RESULTS: The arrival time, peak time, and enhancement time in the group with a nonperfused volume ratio of 70% or higher were longer than those in the group with a volume ratio lower than 70% (mean ± SD, 16.7 ± 3.5, 26.5 ± 4.9, and 10.2 ± 2.6 seconds, respectively, versus 13.3 ± 4.2, 20.8 ± 5.4, and 7.6 ± 2.3 seconds), whereas patients with a volume ratio of 70% or higher had a lower mean enhancement intensity and enhancement rate than those with a volume ratio lower than 70% (29.7 ± 16.7 dB and 3.2 ± 1.5 dB/s versus 63.2 ± 26.3 dB and 8.6 ± 4.3 dB/s; P < .05). The nonperfused volume ratio was negatively correlated with the enhancement intensity and enhancement rate (r = -0.631 and -0.712) but positively correlated with the arrival time, peak time, and enhancement time (r = 0.322, 0.456, and 0.477; P < .05). The areas under the receiver operating characteristic curve for the enhancement time, enhancement intensity, and enhancement rate were 0.73, 0.79, and 0.81 (P < .05). CONCLUSIONS: Quantitative parameters from CEUS are potentially useful for evaluating the therapeutic effect of HIFU ablation for uterine fibroids.

A novel route for the removal of bodily heavy metal lead (II).

The lead ion concentration in bile is considerably higher than in blood, and bile is released into the alimentary tract. Thiol-modified SBA-15 administered orally can combine with lead ions in the alimentary tract. In this paper, the in vitro lead absorption of bile was investigated. This thiol-modified SBA-15 material was used in pharmacodynamics studies on rabbits. The result that the lead content in faeces was notably higher indicates that thiol-modified SBA-15 can efficiently remove lead. The mechanism could include the following: thiol-modified SBA-15 material cuts off the heavy metal lead recirculation in the process of bile enterohepatic circulation by chelating the lead in the alimentary tract, causing a certain proportion of lead to be removed by the thiol mesoporous material, and the lead is subsequently egested out of the body in faeces. The results indicate that this material might be a potential non-injection material for the removal bodily heavy metal lead in the alimentary tract. This material may also be a useful means of lead removal, especially for non-acute sub-poisoning symptoms.

Transcriptome analysis reveals PRKCA as a potential therapeutic target for overcoming cisplatin resistance in lung cancer through ferroptosis.

Cisplatin-based chemotherapy is the current standard care for lung cancer patients; however, drug resistance frequently develops during treatment, thereby limiting therapeutic efficacy.The molecular mechanisms underlying cisplatin resistance remain elusive. In this study, we conducted an analysis of microarray data from the Gene Expression Omnibus (GEO) database under the accession numbers GSE21656, which encompassed expression profiling of cisplatin-resistant H460 (DDP-H460)and the parental cells (H460). Subsequently, we calculated the differentially expressed genes (DEGs) between DDP-H460 and H460. Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of DEGs demonstrated significant impact on the Rap1, PI3K/AKT and MAPK signaling pathways. Moreover, protein and protein interaction (PPI) network analysis identified PRKCA, DET1, and UBE2N as hub genes that potentially contribute predominantly to cisplatin resistance. Ultimately, PRKCA was selected for validation due to its significant prognostic effect, which predicts unfavorable overall survival and disease-free survival in patients with lung cancer. Network analysis conducted on The Cancer Genome Atlas (TCGA) database revealed a strong gene-level correlation between PRKCA and TP53, CDKN2A, BYR2, TTN, KRAS, and PIK3CA; whereas at the protein level, it exhibited a high correlation with EGFR, Lck, Bcl2, and Syk. The in vitro experiments revealed that PRKCA was upregulated in the cisplatin-resistant A549 cells (DDP-A549), while knockdown of PRKCA increased DDP-A549 apoptosis upon cisplatin treatment. Moreover, we observed that PRKCA knockdown attenuated DDP-A549 proliferation, migration and invasion ability. Western blot analysis demonstrated that PRKCA knockdown downregulated phosphorylation of PI3K expression while upregulated the genes involved in ferroptosis signaling. In summary, our results elucidate the role of PRKCA in acquiring resistance to cisplatin and underscore its potential as a therapeutic target for cisplatin-resistant lung cancer.

The value of synthetic MRI in detecting the brain changes and hearing impairment of children with sensorineural hearing loss.

Introduction: Sensorineural hearing loss (SNHL) can arise from a diverse range of congenital and acquired factors. Detecting it early is pivotal for nurturing speech, language, and cognitive development in children with SNHL. In our study, we utilized synthetic magnetic resonance imaging (SyMRI) to assess alterations in both gray and white matter within the brains of children affected by SNHL. Methods: The study encompassed both children diagnosed with SNHL and a control group of children with normal hearing {1.5-month-olds (n = 52) and 3-month-olds (n = 78)}. Participants were categorized based on their auditory brainstem response (ABR) threshold, delineated into normal, mild, moderate, and severe subgroups.Clinical parameters were included and assessed the correlation with SNHL. Quantitative analysis of brain morphology was conducted using SyMRI scans, yielding data on brain segmentation and relaxation time.Through both univariate and multivariate analyses, independent factors predictive of SNHL were identified. The efficacy of the prediction model was evaluated using receiver operating characteristic (ROC) curves, with visualization facilitated through the utilization of a nomogram. It's important to note that due to the constraints of our research, we worked with a relatively small sample size. Results: Neonatal hyperbilirubinemia (NH) and children with inner ear malformation (IEM) were associated with the onset of SNHL both at 1.5 and 3-month groups. At 3-month group, the moderate and severe subgroups exhibited elevated quantitative T1 values in the inferior colliculus (IC), lateral lemniscus (LL), and middle cerebellar peduncle (MCP) compared to the normal group. Additionally, WMV, WMF, MYF, and MYV were significantly reduced relative to the normal group. Additionally, SNHL-children with IEM had high T1 values in IC, and LL and reduced WMV, WMF, MYV and MYF values as compared with SNHL-children without IEM at 3-month group. LL-T1 and WMF were independent risk factors associated with SNHL. Consequently, a prediction model was devised based on LL-T1 and WMF. ROC for training set, validation set and external set were 0.865, 0.806, and 0.736, respectively. Conclusion: The integration of T1 quantitative values and brain volume segmentation offers a valuable tool for tracking brain development in children affected by SNHL and assessing the progression of the condition's severity.

[Cross-sectional study on ankle sprain and its related factors in physical education college].

OBJECTIVE: To explore prevalence, risk factors and treatment of ankle sprain of young college student , in order to obtain accurate epidemiological data. METHODS: From March 2019 to May 2019, 552 college students(1 104 sides of anke joints) from Xi'an Physical Education university were enrolled in study according to inclusion and excludion standard, including 309 males and 243 females aged from 16 to 24 years old with an average of (20.9±3.7) years old. Age, gender, and body mass indes(BMI) etc were recorded. Morbidity of acute and chronic ankle sprains of physical students, treatment after the first sprain (cold compress, cast or plaster bracing and medicine), visual analogue scale (VAS) during walking were assessed through ankle sprain questionnaire;Cumberland ankle instability tool (CAIT), Maryland foot score were applied to assess ankle function. Lateral ankle ligament injury was objectively assessed by musculoskeletal ultrasonography. RESULTS: The prevalence of acute ankle sprain(AAS) was 96.20% (531/552), and the incidence of AAS was 59.96% (622/1 104). The prevalence of chronic ankle joint instability(CAI) was 16.85% (93/552), and the incidence of CAI was 8.97% (99/1 104). In the four categories of sports, college student suffered from multiple sprains in performance majors group was 22.20% (14/63), including of aerobicsand dance performance. The incidence of AAS of ball sports was 8.60%(14/163). After the first sprain, most college students(94.4%) were received cold compression, about 60% of them went to hospital;however, only 44.7% students were received standard treatmens(cast or plaster), only 35.3% of them were received hard ankle orthosis. In 552 college students, 44 students were suffered from more than 4 times of ankle sprain, and the total incidence was 7.97% (44/552). Cumberland score was 26.6±2.4, Cumberland score of students sprained ankle joint more than 4 times was (29.2±1.1), suggested it was a risk factor for ankle joint instability. VAS of students sprained ankle joint more than 4 times was higher than that of less than 4 times(P<0.05), Maryland foot score was significantly lower than that of that of <4 times(P<0.05). Musculoskeletal ultrasonography measured the thickness of anterior tibiofibular ligament(ATFL) was (2.41±0.41) mm, and the thickness of calcaneofibular ligament(CFL) was (1.92±0.21) mm, and had no statistical difference(P>0.05). CONCLUSION: Ninty-four percent college students had at least once ankle sprain, ankle sprains were more common in erobics and ball sports. After the first sprain, the proportion of cast or plaster treatment was less than 50%. Sprained ankle joint more than 4 times is a risk factor, and musculoskeletal ultrasonography showed thickening of both ATFL and CFL, while no statstical difference.

Improved Differential Diagnosis Based on BI-RADS Descriptors and Apparent Diffusion Coefficient for Breast Lesions: A Multiparametric MRI Analysis as Compared to Kaiser Score.

RATIONALE AND OBJECTIVES: To develop the nomogram utilizing the American College of Radiology BI-RADS descriptors, clinical features, and apparent diffusion coefficient (ADC) to differentiate benign from malignant breast lesions. MATERIALS AND METHODS: A total of 341 lesions (161 malignant and 180 benign) were included. Clinical data and imaging features were reviewed. Univariable and multivariable logistic regression analyses were performed to determine the independent variables. ADC as a continuous or classified into binary form with a cutoff value of 1.30 × 10-3 mm2/s, incorporated other independent predictors to construct two nomograms, respectively. Receiver operating curve and calibration plot was employed to test the models' discriminative ability. The diagnostic performance between the developed model and the Kaiser score (KS) was also compared. RESULTS: In both models, high patient age, the presence of root sign, time-intensity curves (TICs) types (plateau and washout), heterogenous internal enhancement, the presence of peritumoral edema, and ADC were independently associated with malignancy. The AUCs of two multivariable models (AUC, 0.957; 95% CI: 0.929-0.976 and AUC, 0.958; 95% CI: 0.931-0.976) were significantly higher than that of the KS (AUC, 0.919, 95% CI: 0.885-0.946; both P < 0.001). At the same sensitivity of 95.7%, our models showed an increase in specificity by 5.56% (P = 0.076) and 6.11% (P = 0.035), respectively, as compared to the KS. CONCLUSION: The models incorporating MRI features (root sign, TIC, margins, internal enhancement, and presence of edema), quantitative ADC value, and patient age showed improved diagnostic performance and might have avoided more unnecessary biopsies in comparison with the KS, although further external validation is required.

MYC Promotes LDHA Expression through MicroRNA-122-5p to Potentiate Glycolysis in Hepatocellular Carcinoma.

MYC is a notorious oncogene in a vast network of malignancies, whereas liver-specific microRNA- (miR-) 122-5p is downregulated in hepatocellular cancer (HCC). Here, we studied the possible correlation between these two and their involvement in glycolysis in HCC. MYC was overexpressed in HCC tissues and cells compared to normal liver tissues and normal hepatocytes NHC, which predicted a poor survival of HCC sufferers. Functional assays demonstrated that silencing of MYC inhibited the glycolysis in HCC cells, as evidenced by significantly weaker glucose consumption, lactate production, adenosine triphosphate (ATP) levels, and downregulated HK1 and HK2 protein expression. Moreover, MYC bound to the miR-122-5p promoter and repressed the miR-122-5p expression. Rescue experiments showed that miR-122-5p inhibitor rescued the diminished glycolysis after MYC silencing. In addition, lactate dehydrogenase (LDHA) was identified as a downstream target of miR-122-5p. The overexpression of LDHA mitigated the effects of si-MYC and miR-122-5p mimic on glycolysis of HCC cells, respectively. In conclusion, the MYC/miR-122-5p/LDHA axis modulates glycolysis in HCC cells and possibly affects HCC progression.

Atractylenolide-1 Targets FLT3 to Regulate PI3K/AKT/HIF1-α Pathway to Inhibit Osteogenic Differentiation of Human Valve Interstitial Cells.

Atractylenolide-1 (AT-1), a natural active ingredient extracted from Atractylodes macrocephala, was reported to have good anti-fibrotic and anti-inflammatory effects. Osteogenic changes induced by the inflammation of valve interstitial cells (VICs) play a role in the development of calcified aortic valve disease (CAVD). This study aimed to investigate the anti-osteogenic effects of AT-1 in human VICs. Human VICs were exposed to osteogenic induction medium (OM) containing AT-1 to analyze cell viability, as well as protein and osteogenic gene expression. Anti-calcification tests were also performed. mRNA transcriptome sequencing was performed to identify differential genes and pathways regulated by AT-1. Western blotting was used to verify the enrichment pathway, protein-protein interaction (PPI) analysis was conducted to identify drug targets. Finally, molecular docking and inhibitors are used to verify the drug targets. Treatment of VICs with 20 µM AT-1 resulted in no significant cytotoxicity. The addition of AT-1 to OM prevented the accumulation of calcified nodules, and decreases in the level of (Alkaline Phosphatase) ALP and RUNX2 gene and protein expression were observed. Atractylenolide-1 can target FLT3 protein and inhibit the phosphorylation of FLT3, thereby blocking PI3K/AKT pathway activation, reducing the production of Hypoxia inducible factor(HIF)1-α, and inhibiting the osteogenic differentiation of VICs. These results suggest AT-1 as a potential drug for treating calcified aortic valve disease.

An LC-MS/MS method for the simultaneous determination of 18 antibacterial drugs in human plasma and its application in therapeutic drug monitoring.

Antimicrobial resistance (AMR) is a major threat to global health due to the wide use of antibacterial drugs. Multiple studies show that the pharmacokinetic/pharmacodynamic (PK/PD) studies of antibiotics are an approach to prevent/delay AMR. The pharmacokinetic parameters of antibiotics are the basis of PK/PD studies, and therapeutic drug monitoring (TDM) is the key method to obtain pharmacokinetic information. We developed an ultra-performance liquid chromatography-tandem mass spectrometry to determine 18 antibacterial drugs (piperacillin, cefazolin, cefuroxime, cefoperazone, ceftriaxone, cefepime, aztreonam, meropenem, imipenem, levofloxacin, moxifloxacin, azithromycin, clindamycin, tigecycline, linezolid, vancomycin, voriconazole and caspofungin) in human plasma for practical clinical usage. Samples were prepared using protein precipitation with methanol. Chromatographic separation was accomplished in 6 min on a BEH C18 column (2.1 × 100 mm, 1.7 µm) using a gradient elution of acetonitrile and 0.1% formic acid in water at a flow rate of 0.3 ml/min. The electrospray ionization source interface was operated in the positive and negative ionization modes. Inter- and intra-day precision, accuracy, recovery, matrix effect, and stability were validated according to the Food and Drug Administration guidance. The correlation coefficients of calibration curves were all greater than 0.99. The accuracies of the 18 antibacterial drugs ranged from 89.1% to 112.4%. The intra-day precision of the analytes ranged from 1.4% to 9.3% and the inter-day precision from 2.1% to 7.2%. The matrix effects ranged from 93.1% to 105.8% and the extraction recoveries ranged between 90.1% and 109.2%. The stabilities of the 18 antibacterial drugs in plasma were evaluated by analyzing three different concentrations following storage at three storage conditions. All samples displayed variations less than 15.0%. The validated method was successfully applied to routine clinical TDM for 231 samples.

Engineering BspQI nicking enzymes and application of N.BspQI in DNA labeling and production of single-strand DNA.

BspQI is a thermostable Type IIS restriction endonuclease (REase) with the recognition sequence 5'GCTCTTC N1/N4 3'. Here we report the cloning and expression of the bspQIR gene for the BspQI restriction enzyme in Escherichia coli. Alanine scanning of the BspQI charged residues identified a number of DNA nicking variants. After sampling combinations of different amino acid substitutions, an Nt.BspQI triple mutant (E172A/E248A/E255K) was constructed with predominantly top-strand DNA nicking activity. Furthermore, a triple mutant of BspQI (Nb.BspQI, N235A/K331A/R428A) was engineered to create a bottom-strand nicking enzyme. In addition, we demonstrated the application of Nt.BspQI in optical mapping of single DNA molecules. Nt or Nb.BspQI-nicked dsDNA can be further digested by E. coli exonuclease III to create ssDNA for downstream applications. BspQI contains two potential catalytic sites: a top-strand catalytic site (Ct) with a D-H-N-K motif found in the HNH endonuclease family and a bottom-strand catalytic site (Cb) with three scattered Glu residues. BlastP analysis of proteins in GenBank indicated a putative restriction enzyme with significant amino acid sequence identity to BspQI from the sequenced bacterial genome Croceibacter atlanticus HTCC2559. This restriction gene was amplified by PCR and cloned into a T7 expression vector. Restriction mapping and run-off DNA sequencing of digested products from the partially purified enzyme indicated that it is an EarI isoschizomer with 6-bp recognition, which we named CatHI (CTCTTC N1/N4).

The Pharmacokinetics and Tissue Distributions of Nine Steroidal Saponins from Paris polyphylla in Rats.

BACKGROUND AND OBJECTIVES: Paris polyphylla (P. polyphylla) is a herb widely used in traditional Chinese medicine to treat various diseases. This study used ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to study the pharmacokinetics and tissue distributions of nine steroidal saponins from P. polyphylla. METHODS: P. polyphylla extract was administered to rats intravenously (i.v.) and orally (p.o.). The concentrations of the nine main bioactive components of the extract were determined in plasma and tissue samples using UPLC-MS/MS. The nine saponin compounds were also incubated in an anaerobic environment with intestinal flora suspension solution to investigate hydrolysis by intestinal flora. RESULTS: After oral administration of the P. polyphylla extract, polyphyllin VII was found to have the highest maximum concentration (Cmax, 17.0 ± 2.24 µg/L) of all nine components, followed by the Cmax values of dioscin (16.17 ± 0.64 µg/L) and polyphyllin H (11.75 ± 1.28 µg/L), while the Cmax values of polyphyllin I, polyphyllin II, progenin III, polyphyllin IV, gracillin, and polyphyllin were less than 10 µg/L. The bioavailabilities of all nine components were less than 1%. All the compounds were hydrolyzed by intestinal flora and were predominantly distributed in the liver and lungs. CONCLUSIONS: The nine compounds presented different pharmacokinetic parameter values, and multiple administrations did not accumulate in the body. The bioavailabilities of the compounds were low, partly because of hydrolysis by intestinal flora. The nine compounds were mainly distributed in the liver and lungs, which may be target organs.

Degradation of Rhodamine B in aqueous solution by laser cavitation.

A novel method of laser cavitation (LC) was proposed for degrading organic dye wastewater. Rhodamine B (RhB) aqueous solution was employed as the simulated organic dye wastewater, and a LC system was designed to conduct the experiments of degrading RhB. The effects of laser energy, initial concentration and cavitation time on the degradation were investigated. Moreover, the degradation kinetics, degradation mechanism and energy efficiency were analyzed. The experimental results indicate that RhB aqueous solution can be degraded effectively by LC and the degradation follows the pseudo-first-order kinetics. The extent of degradation increases by 27.6% with the rise of laser energy (50-100 mJ) while it decreases by 7.8% with increasing the initial concentration from (20-40 mg/L), but RhB can not be degraded when exceeding 100 mg/L. The degradation extent of RhB at 100 mJ and 20 mg/L for 3 h is 81.11%, and the RhB solution is almost completely degraded at 150 mJ (98.4%). The degradation velocity of RhB rises firstly and then decreases as the cavitation time increases. The degradation of RhB by LC can be attributed to the N-de-ethylation and chromophore cleavage caused by oxidation of hydroxyl (OH) radical and thermal decomposition. LC has a higher energy efficiency compared with other methods and is more energy efficient at lower laser energy.

Discovery of natural nicking endonucleases Nb.BsrDI and Nb.BtsI and engineering of top-strand nicking variants from BsrDI and BtsI.

BsrDI and BtsI restriction endonucleases recognize and cleave double-strand DNA at the sequences GCAATG (2/0) and GCAGTG (2/0), respectively. We have purified and partially characterized these two enzymes, and analyzed the genes that encode them. BsrDI and BtsI are unusual in two respects: each cleaves DNA as a heterodimer of one large subunit (B subunit) and one small subunit (A subunit); and, in the absence of their small subunits, the large subunits behave as sequence-specific DNA nicking enzymes and only nick the bottom strand of the sequences at these respective positions: GCAATG (-/0) and GCAGTG (-/0). We refer to the single subunit, the bottom-strand nicking forms as 'hemidimers'. Amino acid sequence comparisons reveal that BsrDI and BtsI belong to a family of restriction enzymes that possess two catalytic sites: a canonical PD-X(n)-EXK and a second non-canonical PD-X(n)-E-X12-QR. Interestingly, the other family members, which include BsrI (ACTGG 1/-1) and BsmI/Mva1269I (GAATGC 1/-1) are single polypeptide chains, i.e. monomers, rather than heterodimers. In BsrDI and BtsI, the two catalytic sites are found in two separate subunits. Site-directed mutagenesis confirmed that the canonical catalytic site located at the N-terminus of the large subunit is responsible for the bottom-strand cleavage, whereas the non-canonical catalytic site located in the small subunit is responsible for hydrolysis of the top strand. Top-strand specific nicking variants, Nt.BsrDI and Nt.BtsI, were successfully engineered by combining the catalytic-deficient B subunit with wild-type A subunit.

Characterization of transcriptome profile and clinical features of a novel immunotherapy target CD204 in diffuse glioma.

CD204 is a specific marker of tumor-associated macrophages (TAMs) in glioma. However, the expression levels of CD204 and its involvement in glioma are not fully understood. In this large-scale study, we assessed the expression and function of CD204 in whole-grade glioma molecularly and clinically. In total, 1323 glioma samples, including 301 microarray data and 325 RNA-seq data from the Chinese Glioma Genome Atlas (CGGA) dataset and 697 RNA-seq data from The Cancer Genome Atlas (TCGA) dataset, were utilized. The statistical analysis and graphical work were mainly performed using the R software. Univariate and multivariate Cox analysis demonstrated that CD204 was an independent prognosticator in glioma patients. CD204 expression was positively correlated with the grade of malignancy. CD204 was consistently upregulated in wild-type isocitrate dehydrogenase glioma and highly expressed in mesenchymal glioblastoma. Gene ontology of CD204-related genes showed that CD204 was most enriched in inflammatory response and immune response. It was associated with the stromal and immune populations, especially the monocytic lineage, fibroblasts, and T cells. Circos plots revealed that CD204 was closely associated with many immune checkpoint regulators, especially TIM-3. CD204 expression is consistent with the malignant phenotype of glioma and independently predicts poor outcomes in glioma patients. Additionally, CD204+ TAMs, collaborating with other checkpoint members, may contribute to the dysfunction of T cells. These findings suggest that CD204 may be a promising target for glioma immunotherapy.

A functional intronic variant in the tyrosine hydroxylase (TH) gene confers risk of essential hypertension in the Northern Chinese Han population.

The TH (tyrosine hydroxylase) gene encodes the rate-limiting enzyme of catecholamine biosynthesis, and is involved in the pathogenesis of hypertension, but the relationship of its variants with hypertension has not been extensively studied. We designed a case-controlled study consisting of 503 HT (hypertensive) individuals and 490 NT (normotensive) individuals matched by region, age and gender to systematically investigate the association between the TH gene and hypertension. Based on the HapMap and dbSNP (where SNP is single nucleotide polymorphism) data, four SNPs, rs6356 A>G, rs6357 G>A, rs2070762 T>C and rs1800033 A>G in the TH gene were selected for genotyping. Rs1800033 was not polymorphic in our study population. No significant differences were observed for distributions of rs6356 and rs6357 between the HT and NT groups. However, both the genotype and allele frequencies of rs2070762 showed significant differences between cases and controls (P<0.001 and P=0.005 respectively). In haplotype analysis, a total of eight haplotypes were observed in the entire population and the overall frequency distributions differed significantly between the HT and NT groups. Specifically, haplotype A-A-C (rs6356-rs6357-rs2070762) occurred only in the HT group and A-G-C occurred more commonly in HT subjects than in NT subjects (P=0.003 and P=0.013 respectively). Compared with the most common haplotype A-G-T, the adjusted OR (odds ratio) was 1.83 [95% CI (confidence interval), 1.20-2.79; P=0.0049] for haplotype G-G-C and 20 (P<0.0001) for the haplotype A-A-C. Functional analysis showed that the C allele of rs2070762 functioned as an enhancer in the absence of binding by unidentified transcriptional repressor(s). These results provide evidence for an association of the functional intronic rs2070762 with essential hypertension.

Renalase gene is a novel susceptibility gene for essential hypertension: a two-stage association study in northern Han Chinese population.

Renalase, a novel flavin adenine dinucleotide-dependent amine oxidase, is secreted by the kidney, degrades circulating catecholamines, and modulates cardiac function and systemic blood pressure (BP). Its discovery may provide novel insights into the mechanisms of BP regulation and the pathogenesis of essential hypertension (EH). We designed a two-stage case-control study to investigate whether the renalase gene harbored any genetic variants associated with EH in the northern Han Chinese population. From the International Collaborative Study of Cardiovascular Disease in Asia (InterASIA in China), 1,317 hypertensive cases and 1,269 normotensive controls were recruited. These total 2,586 subjects were taken as the main study population in this study. In stage 1, all the eight selected single nucleotide polymorphisms (SNPs) of the renalase gene were genotyped and tested within a subsample (503 cases and 490 controls) of the main study population. By single locus analyses, three SNPs, rs2576178, rs2296545, and rs2114406, showed significant associations with EH (P < 0.05). In stage 2, these three SNPs were genotyped on the remaining individuals and analyzed using all the individuals. After Bonferroni correction for multiple comparisons, the associations of rs2576178 and rs2296545 with EH were still significant in stage 2. The cases had higher frequencies of rs2576178 G allele and rs2296545 C allele than the controls (0.55 versus 0.49, P < 0.0001; 0.61 versus 0.55, P < 0.0001). Particularly, under the codominant model, the adjusted odds ratios for rs2576178 GG genotype and rs2296545 CC genotype were 1.58 (95% CI, 1.25 to 2.00; P = 0.0002) and 1.61 (95% CI, 1.26 to 2.04; P = 0.0002), respectively. We also found risk-associated haplotypes and diplotypes, which further confirmed the significant association between the renalase gene and EH. These findings may provide novel genetic susceptibility markers for EH and lead to a better understanding of EH pathophysiology. In addition, further replications in other populations and functional studies would be warranted.

Relationship of WNT4 Gene with the Risk of Epithelial Ovarian Cancer: A Han Chinese Population-Based Association Study.

OBJECTIVE: In China, epithelial ovarian cancer (EOC) patients account for the majority of ovarian cancer patients. The pathogenesis of EOC, one of the most lethal gynecological malignancies, remains unclear. Recently, the role of WNT4 in gynecological disease and tumor development was reported, and a suspicious association of WNT4 with EOC was identified in Europeans. However, the contributions of the WNT4 gene to EOC and the underlying molecular mechanisms remains largely unknown. To determine whether the WNT4 gene is associated with EOC, this study investigated polymorphisms of the WNT4 gene in Han Chinese individuals. MATERIALS AND METHODS: We designed a case/control study with 707 EOC patients and 1563 unrelated healthy controls of Han Chinese descent. A total of eight tag single-nucleotide polymorphisms (SNPs) were genotyped successfully, and both single SNP and haplotype analyses were performed to detect the potential association of variations in the WNT4 gene with EOC. RESULTS: The SNP rs56318008 was found to be strongly associated with EOC risk. In the serous EOC subgroup, individuals harboring the T allele of rs56318008 exhibited a higher risk of EOC than individuals harboring the C allele. Moreover, the odds ratios and 95% confidence intervals revealed an increased risk of EOC in individuals with the T allele of the SNP, and haplotypic analyses confirmed the results, showing a similar pattern. CONCLUSION: Our results show that the WNT4 gene is associated with EOC risk, indicating that this gene may be a potential genetic risk factor for developing EOC.

Role of CXCR7 as a Common Predictor for Prognosis in Solid Tumors: a Meta-Analysis.

Background: Accumulating evidence indicated that the CXC chemokine receptor (CXCR) 7 (CXCR7) was overexpressed in a variety of tumors. However, the value of the CXCR7 expression in predicting prognosis in solid tumors remains controversial. Therefore, we performed this meta-analysis to evaluate the correlation between CXCR7 expression and lymph node metastasis (LNM), tumor pathological grade and survival, including overall survival (OS), disease-free survival (DFS) and recurrence-free survival (RFS). Methods: Eligible studies were searched in PubMed, Web of Science, and PMC up to April 2018. A total of 27 studies were included in this meta-analysis. Odds ratio (OR), hazard ratio (HR) and 95 % confidence intervals (CI) were used as effect measures. Results: The meta-analysis showed that high expression of CXCR7 predicted a high risk of LNM (pooled OR = 2.22, 95%CI: 1.41-3.50), high tumor grade (pooled OR = 1.94, 95%CI: 1.20-3.13), poor OS (pooled HR = 1.66, 95%CI: 1.30-2.03), and poor DFS/RFS (pooled HR = 1.82, 95%CI: 1.21-2.43). Subgroup analysis showed that CXCR7 expression had a positive correlation with LNM in pan-adenocarcinoma subgroup (pooled OR = 3.73, 95%CI: 2.21-6.30), while no correlation was found in pan-squamous cancer subgroup (pooled OR = 1.29, 95%CI: 0.56-2.96). Subgroup analysis of tumor grade revealed that high expression of CXCR7 predicted high tumor grade both in pan-squamous cancer and pan-adenocarcinoma (pooled OR = 3.58, 95%CI: 1.39-9.22, pooled OR = 2.25, 95%CI: 1.20-4.20). As in OS group, we divided the data based on analysis method and it turned out that overexpressed CXCR7 predicted worse OS both in multivariate analysis (pooled HR =1.57, 95%CI: 1.12-2.01) and univariate analysis subgroup (pooled HR =1.86, 95%CI: 1.23-2.49). Conclusions: Our meta-analysis revealed that high expression of CXCR7 predicted unfavorable prognosis and may serve as potential targets of cancer therapy.

Rational design of a chimeric endonuclease targeted to NotI recognition site.

A cleavage-deficient variant of NotI restriction endonuclease (GCGGCCGC) was isolated by random mutagenesis of the notIR gene. The NotI variant D160N was shown to bind DNA and protect plasmid DNA from EagI (CGGCCG) and NotI digestions. The EDTA-resistant BmrI restriction endonuclease cleaves DNA sequence ACTGGG N5/N4. The N-terminal cleavage domain of BmrI (residues 1-198) with non-specific nuclease activity was fused to the NotI variant D160N with a short linker. The engineered chimeric endonuclease (CH-endonuclease) recognizes NotI sites specifically in the presence of high salt (100-150 mM NaCl) and divalent cations Mg++ or Ca++. In contrast to wild-type NotI, which cuts within its recognition sequence, BmrI198-NotI (D160N) cleaves DNA outside of NotI sites, resulting in deletion of the NotI site and the adjacent sequences. The fusion of the BmrI cleavage domain to cleavage-deficient variants of Type II restriction enzymes to generate novel cleavage sites will provide useful tools for DNA manipulation.