• 제목/요약/키워드: Single cell analysis/methods

검색결과 203건 처리시간 0.032초

Reciprocal regulation of SIRT1 and AMPK by Ginsenoside compound K impedes the conversion from plasma cells to mitigate for podocyte injury in MRL/lpr mice in a B cell-specific manner

  • Ziyu Song;Meng Jin;Shenglong Wang;Yanzuo Wu;Qi Huang;Wangda Xu;Yongsheng Fan;Fengyuan Tian
    • Journal of Ginseng Research
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    • 제48권2호
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    • pp.190-201
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    • 2024
  • Background: Deposition of immune complexes drives podocyte injury acting in the initial phase of lupus nephritis (LN), a process mediated by B cell involvement. Accordingly, targeting B cell subsets represents a potential therapeutic approach for LN. Ginsenoside compound K (CK), a bioavailable component of ginseng, possesses nephritis benefits in lupus-prone mice; however, the underlying mechanisms involving B cell subpopulations remain elusive. Methods: Female MRL/lpr mice were administered CK (40 mg/kg) intragastrically for 10 weeks, followed by measurements of anti-dsDNA antibodies, inflammatory chemokines, and metabolite profiles on renal samples. Podocyte function and ultrastructure were detected. Publicly available single-cell RNA sequencing data and flow cytometry analysis were employed to investigate B cell subpopulations. Metabolomics analysis was adopted. SIRT1 and AMPK expression were analyzed by immunoblotting and immunofluorescence assays. Results: CK reduced proteinuria and protected podocyte ultrastructure in MRL/lpr mice by suppressing circulating anti-dsDNA antibodies and mitigating systemic inflammation. It activated B cell-specific SIRT1 and AMPK with Rhamnose accumulation, hindering the conversion of renal B cells into plasma cells. This cascade facilitated the resolution of local renal inflammation. CK facilitated the clearance of deposited immune complexes, thus reinstating podocyte morphology and mobility by normalizing the expression of nephrin and SYNPO. Conclusions: Our study reveals the synergistic interplay between SIRT1 and AMPK, orchestrating the restoration of renal B cell subsets. This process effectively mitigates immune complex deposition and preserves podocyte function. Accordingly, CK emerges as a promising therapeutic agent, potentially alleviating the hyperactivity of renal B cell subsets during LN.

Color discrepancy of single-shade composites at different distances from the interface measured using cell phone images

  • Marcia Luciana Carregosa Santana;Gabriella de Jesus Santos Livi;Andre Luis Faria-e-Silva
    • Restorative Dentistry and Endodontics
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    • 제49권1호
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    • pp.7.1-7.11
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    • 2024
  • Objectives: This study aimed to evaluate the impact of substrate color and interface distance on the color adjustment of 2 single-shade composites, Vittra APS Unique and Charisma Diamond One. Materials and Methods: Dual disc-shaped specimens were created using Vittra APS Unique or Charisma Diamond One as the center composite, surrounded by shaded composites (A1 or A3). Color measurements were taken with a spectrophotometer against a gray background, recording the color coordinates in the CIELAB color space. Illumination with a light-correcting device and image acquisition using a polarizing filter-equipped cell phone were performed on specimens over the same background. Image processing software was used to measure the color coordinates in the center and periphery of the inner composite and in the outer composite. The color data were then converted to CIELAB coordinates and adjusted using data from the spectrophotometer. Color differences (ΔE00) between the center/periphery of single-shade and outer composites were calculated, along with color changes in single-shade composites caused by different outer composites. Color differences for the inner composites surrounded by A1 and A3 were also calculated. Data were analyzed using repeated-measures analysis of variance (α = 0.05). Results: The results showed that color discrepancies were lowest near the interface and when the outer composite was whiter (A1). Additionally, Charisma Diamond One exhibited better color adjustment ability than Vittra APS Unique. Conclusions: Color discrepancies between the investigated single-shade composites diminished towards the interface with the surrounding composite, particularly when the latter exhibited a lighter shade.

Analysis of allele-specific expression using RNA-seq of the Korean native pig and Landrace reciprocal cross

  • Ahn, Byeongyong;Choi, Min-Kyeung;Yum, Joori;Cho, In-Cheol;Kim, Jin-Hoi;Park, Chankyu
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권12호
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    • pp.1816-1825
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    • 2019
  • Objective: We tried to analyze allele-specific expression in the pig neocortex using bioinformatic analysis of high-throughput sequencing results from the parental genomes and offspring transcriptomes from reciprocal crosses between Korean Native and Landrace pigs. Methods: We carried out sequencing of parental genomes and offspring transcriptomes using next generation sequencing. We subsequently carried out genome scale identification of single nucleotide polymorphisms (SNPs) in two different ways using either individual genome mapping or joint genome mapping of the same breed parents that were used for the reciprocal crosses. Using parent-specific SNPs, allele-specifically expressed genes were analyzed. Results: Because of the low genome coverage (${\sim}4{\times}$) of the sequencing results, most SNPs were non-informative for parental lineage determination of the expressed alleles in the offspring and were thus excluded from our analysis. Consequently, 436 SNPs covering 336 genes were applicable to measure the imbalanced expression of paternal alleles in the offspring. By calculating the read ratios of parental alleles in the offspring, we identified seven genes showing allele-biased expression (p<0.05) including three previously reported and four newly identified genes in this study. Conclusion: The newly identified allele-specifically expressing genes in the neocortex of pigs should contribute to improving our knowledge on genomic imprinting in pigs. To our knowledge, this is the first study of allelic imbalance using high throughput analysis of both parental genomes and offspring transcriptomes of the reciprocal cross in outbred animals. Our study also showed the effect of the number of informative animals on the genome level investigation of allele-specific expression using RNA-seq analysis in livestock species.

Single Nucleotide Polymorphisms in the u-PA Gene are Related to Susceptibility to Oral Tongue Squamous Cell Carcinoma in the Northern Chinese Han Population

  • Zhong, Feng;Yang, Xue-Cai;Bu, Ling-Xue;Li, Ning-Yi;Chen, Wan-Tao
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권2호
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    • pp.781-784
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    • 2013
  • Aim: The purpose of this study was to determine whether susceptibility to oral tongue squamous cell carcinoma (OSCC) is related to polymorphisms in the u-PA gene. Methods: We examined the rs2227564 C/T and rs2227562 G/A single nucleotide polymorphisms (SNPs) in 196 OSCC patients and 201 age- and gender-matched controls via direct sequencing and PCR-RFLP methods. Results: Significant differences were found in allelic and genotypic distributions of the rs2227564 and rs2227562 loci when comparing cases and controls. In addition, logistic analyses indicated that the rs2227564 C/T genotype was related to a 1.52-fold increased risk of developing OSCC (adjusted OR=1.521, 95%CI: 1.144~2.022, P=0.004). Linkage disequilibrium analysis was conducted and no association between the two loci was found (D'=0.031, $r^2$=0.000). Conclusions: Our findings provide evidence that the rs2227564 C/T SNP in the u-PA gene is associated with the development of OSCC.

환경 오염물질의 진보된 독성 평가 기법 (Recent Advanced Toxicological Methods for Environmental Hazardous Chemicals)

  • 류재천;최윤정;김연정;김형태;방형애;송윤선
    • Environmental Analysis Health and Toxicology
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    • 제14권1_2호
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    • pp.1-12
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    • 1999
  • Recently, several new methods for the detection of genetic damages in vitro and in vivo based on molecular biological techniques were introduced according to the rapid progress in toxicology combined with cellular and molecular biology. Among these methods, mouse lymphoma thymidine kanase (tk) gene forward mutation assay, single cell gel electrophoresis (comet assay) and transgenic animal and cell line model as a target gene of lac I (Big Blue) and lac Z (Muta Mouse) gene mutation are newly introduced based on molecular toxicological approaches. The mouse lymphoma tk$\^$+/-/ gene assay (MOLY) using L5178Y tk$\^$+/-/ mouse lymphoma cell line is one of the mammalian forward mutation assays, and has many advantages and more sensitive than hprt assay. The target gene of MOLY is a heterozygous tk$\^$+/-/ gene located in 11 chromosome, so it is able to detect the wide range of genetic changes like point mutation, deletion, rearrangement, and mitotic recombination within tk gene or deletion of entire chromosome 11. The comet assay is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakages in mammalian cells, Also, transgenic animal and cell line models, which have exogenous DNA incorporated into their genome, carry recoverable shuttle vector containing reporter genes to assess endogenous effects or alteration in specific genes related to disease process, are powerful tools to study the mechanism of mutation in vivo and in vitro, respectively. Also in vivo acridine orange supravital staining micronucleus assay by using mouse peripheral reticulocytes was introduced as an alternative of bone marrow micronucleus assay. In this respect, there was an International workshop on genotoxicity procedure (IWGTP) supported by OECD and EMS (Environmental Mutagen Society) at Washington D. C. in March 25-26, 1999. The objective of IWGTP is to harmonize the testing procedures internationally, and to extend to finalization of OECD guideline, and to the agreement of new guidelines under the International Conference of Harmonization (ICH) for these methods mentioned above. Therefore, we introduce and review the principle, detailed procedure, and application of MOLY, comet assay, transgenic mutagenesis assay and supravital staining micronucleus assay.

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Keratinization of Lung Squamous Cell Carcinoma Is Associated with Poor Clinical Outcome

  • Park, Hye Jung;Cha, Yoon-Jin;Kim, Seong Han;Kim, Arum;Kim, Eun Young;Chang, Yoon Soo
    • Tuberculosis and Respiratory Diseases
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    • 제80권2호
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    • pp.179-186
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    • 2017
  • Background: Although the World Health Organization (WHO) classification of lung squamous cell carcinoma (SCC) was revised in 2015, its clinical implications for lung SCC subsets remain unclear. We investigated whether the morphologic characteristics of lung SCC, including keratinization, were associated with clinical parameters and clinical outcome of patients. Methods: A total of 81 patients who underwent curative surgical resection of diagnosed lung SCC, were enrolled in this study. Attributes such as keratinization, tumor budding, single cell invasion, and nuclear size within the tumor, as well as immunohistochemistry of Bcl-xL and pS6 expressions, were evaluated. Results: The keratinizing and nonkeratinizing subtypes did not differ with respect to age, sex, TNM stage, and morphologic parameters such as nuclear diameter, tumor budding, and single cell invasion at the tumor edge. Most patients with the keratinizing subtype (98.0%) had a history of smoking, whereas the nonkeratinizing group had a relatively higher proportion of never-smokers relative to the keratinizing group (24.0% vs. 2.0%; p=0.008, chi-square test). Expression of pS6 (a surrogate marker of mammalian target of rapamycin complex 1 [mTORC1] signaling that regulates keratinocyte differentiation), and Bcl-xL (a key anti-apoptotic molecule that may inhibit keratinization), did not correlate significantly with the presence of keratinization. Patients with the keratinizing subtype had a significantly shorter overall survival (85.2 months vs. 135.7 months, p=0.010, log-rank test), and a multivariate analysis showed that keratinization was an independent, poor prognostic factor (hazard ratio, 2.389; 95% confidence interval, 1.090-5.233; p=0.030). Conclusion: In lung SCC, keratinization is associated with a poor prognosis, and might be associated with smoking.

ATM 전송망에서의 PBS를 이용한 셀 우선 순위 제어 방식의 연구 (An Analysis of Cell Loss Process in an ATM Network Under Partial Buffer Sharing Policy)

  • 곽민곤;성수란;김종권
    • 한국통신학회논문지
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    • 제19권12호
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    • pp.2328-2339
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    • 1994
  • ATM 전송 기술에 있어 PBS 방법은 혼재된 트래픽 환경하에서 각각의 트래픽이 서로 다른 서비스 품질을 요구할 경우, 체증 상태의 망 관리를 효율적으로 하기 위한 제시된 셀 준위의 우선 순위 제어방식이다. 이 논문에서는 시간 지연과 셀 손실에 민감한 두 트래픽에 대한 처리를 PBS 방법을 적용시켜 셀 손실에 대한 분포를 분석하고자 한다. 특히, 실시간 트래픽의 주요한 성능 척도인 연속적 셀 손실 확률을 정확하게 구하기 위한 셀 손실 간의 독립성을 가정하지 않은 재귀적 알고리듬을 사용한다. 이 방법에 의해 유도된 셀 손실에 대한 분포의 결과는 독립을 가정한 경우보다 더 심각하게 나타난다. 그리고, 실시간 트래픽에 대한 PBS 방법의 제한성과 주어진 서비스 품질 기준에 따른 허용 부하를 증가시킬 수 있음을 보여준다.

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Effects on hematology and blood biochemistry profile of intramuscular meloxicam injection in Brahminy kite and Barn owl

  • Ratiwan Sitdhibutr;Raveewan Ploypan;Sirawit Subaneg;Chaiyan Kasorndorkbua
    • Journal of Veterinary Science
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    • 제24권3호
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    • pp.43.1-43.8
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    • 2023
  • Background: Meloxicam is used widely for exotic animal analgesia, but its toxicity in common raptor species in Thailand is unclear. Objectives: This study evaluated the single-dose effect of intramuscular meloxicam in common raptor species in Thailand for short-term and long-term periods. Methods: Twenty-two raptors were administered a single 1 mg/kg dose of meloxicam individually via intramuscular injection. The following were evaluated: clinical appearance, body weight, body condition score, body temperature, fecal appearance, complete blood cell count, and biochemistry panel before (day 0) and after the injection (1, 7, and 30 days). The collected samples were categorized into three groups: Brahminy kite (Haliastur indus) (n = 10), adult Barn owl (Tyto javanica) (n = 4), and juvenile Barn owl (n = 8). Results: None of the raptors in the study groups showed any abnormalities. The hematological profiles were significantly different in the short-term period (day 1 and day 7). The creatinine, aspartate aminotransferase, and creatinine kinase increased in several groups. On the other hand, the packed cell volume decreased in the Brahminy kite and juvenile Barn owl groups. According to the findings, an intramuscular injection of 1 mg/kg meloxicam affected the blood biochemistry panel of the muscle, but the affected raptors recovered within one week. Conclusions: An intramuscular injection of meloxicam at a single 1 mg/kg dose in Brahminy kites and Barn owls was not associated with the morbidity, hepatotoxicity, gastrointestinal toxicity, and nephrotoxicity in the short- and long-term periods.

Two Crystal Structures of Dehydrated Fully $Ca^{2+}$-Exchanged Zeolte A Reacting with Rubidium Vapor

  • Song, Seong-Hwan;Kim, Yang
    • Bulletin of the Korean Chemical Society
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    • 제14권2호
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    • pp.258-262
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    • 1993
  • Two single crystals of fully dehydrated $Rb^+$ -exchanged zeolite A have been prepared by the reduction of all $Ca^{2+}$ ions in dehydrated $Ca_6$-A by rubidium vapor. Their structures were determined by single crystal X-ray diffraction methods in the cubic space group Pm3m (a=12.160(2) $^{\AA}$ and 12.166(2) $^{\AA}$) at 22(1)$^{\circ}$C. In these structures, 12.4(2) to 13.3(2) Rb species are found per unit cell, more than 12 Rb$^+$ ions needed to balance the anionic charge of the zeolite framework, indicating that the sorption $Rb^0$ has occurred. In each structure, three $Rb^+$ ions per unit cell are located at the centers of the 8-rings. Six to eight $Rb^+$ ions are found opposite the 6-rings on threefold axes, and three $Rb^+$ ions are found in a sodalite unit. About 0.5 $Rb^+$ ion lies opposite a 4-ring. The structural analysis indicates the presence of a triangular rubidium cluster in the sodalite cavities. The triangular rubidium clusters may be stabilized by the coordination to two and/or three rubidium ions in the large cavity. Therefore, this cluster may be viewed as $(Rb_5)^{4+}$ and/or $(Rb_6)^{4+}$.

Clinical Observation and Prognostic Analysis of Pemetrexed plus Platinum as First-line Treatment in Patients with Advanced Non-small Cell Lung Cancer

  • Wang, Ji-Ying;Cai, Yong
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권11호
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    • pp.6267-6271
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    • 2013
  • Objective: To determine clinical efficacy, safety and prognostic factors of pemetrexed plus platinum as first-line treatment in patients with advanced non-small cell lung cancer (NSCLC). Materials and Methods: Clinical characteristics, short-term efficacy, survival and adverse reactions of 47 advanced non-squamous NSCLC patients who had received pemetrexed plus platinum as first-line treatment in Shanghai Pulmonary Hospital from January 2009 to June 2011 were retrospectively analyzed. The Chi-squared test was applied to statistically analyze the overall response rate (ORR), disease control rate (DCR) and toxicity reactions in both groups, while survival data wereanalyzed by Kaplan-Meier and logrank methods, and the COX proportional hazards model was adopted for a series of multi-factor analyses. Results: Only two patients were lost to follow-up. The ORR, DCR, medium progression-free survival time (PFS) and medium overall survival (OS) were 31.9%, 74.5%, 5 months and 15.2 months, while 1- and 2-year survival rates were 63.8% (30/47) and 19.2% (9/47), respectively. Single-factor analysis showed that tumor pathological patterns and efficacy were in association with medium PFS (P<0.05), whereas tumor pathological patterns, smoking history and efficacy were closely connected with medium OS (P<0.05). Multi-factor analyses demonstrated that pathological patterns and efficacy were independent factors influencing OS (P<0.05). The rate of toxicity reactions in degree III/IV was low, including hematologic toxicity marked by decline in white blood cell count and decrease in the platelet count (PLT), and non-hematologic toxicity manifested by gastrointestinal reactions, such as nausea and vomiting. Conclusions: Pemetrexed plus platinum as first-line treatment has excellent efficacy and slight adverse reactions with favorable drug-tolerance in patients with advanced non-squamous NSCLC.