Ml Of Peripheral Venous Blood Research Articles

Clinical Characteristics, Genetic Analysis, and Literature Review of Cornelia de Lange Syndrome Type 4 Associated With a RAD21 Variant.

Cornelia de Lange syndrome (CdLS) is an uncommon congenital developmental disorder distinguished by intellectual disorder and distinctive facial characteristics, with a minority of cases attributed to RAD21 variants. A patient was admitted to the endocrinology department at Peking Union Medical College Hospital, where 2 mL of peripheral venous blood was collected from the patient and his parents. DNA was extracted for whole-exome sequencing (WES) analysis, and the genetic variation of the parents was confirmed through Sanger sequencing. A 13.3-year-old male patient with a height of 136.5 cm (-3.5 SDS) and a weight of 28.4 kg (-3.1 SDS) was found to have typical craniofacial features. WES revealed a pathogenic variant c.1143G>A (p.Trp381*) in the RAD21 gene. He was diagnosed with CdLS type 4 (OMIM #614701). We reviewed 36 patients with CdLS related to RAD21 gene variants reported worldwide from May 2012 to March 2024. Patient's variant status, clinical characteristics, and rhGH treatment response were summarized. Frameshift variants constituted the predominant variant type, representing 36% (13/36) of cases. Clinical features included verbal developmental delay and intellectual disorder observed in 94% of patients. This study reported the third case of CdLS type 4 in China caused by a RAD21 gene variant, enriching the genetic mutational spectrum.

Genetic analysis of a novel FBN1 mutation in a pediatric Marfan syndrome patient.

The aim of this study was to investigate a novel FBN1 gene mutation in a pediatric patient with Marfan syndrome (MFS) to provide a theoretical basis for genetic counseling. The subject was a 5-month-old male infant. With informed consent from the proband and his family, 2 mL of peripheral venous blood was collected from the patient, his father, mother, and sister. DNA was extracted using a DNA extraction kit with EDTA-K as an anticoagulant. The extracted DNA was subjected to minigene transcription and bioinformatics analysis. For minigene construction, wild-type and mutant minigenes were inserted into pcMINI and pcMINI-C vectors, respectively. Four recombinant vectors were transfected into the HeLa and 293T cell lines. After transfection for 48 hours, RNA was extracted from eight samples. DNA was also extracted from the family members' samples to construct a library. Target regions were captured using the SureSelect Human All Exon V6 (Agilent) kit and were sequenced with Illumina NovaSeq (sequencing read length 2×150 bp). Bioinformatic analysis identified the c.8226+5del mutation as a variant of uncertain clinical significance (VOUS). Literature and database reviews confirmed that this mutation had not been previously reported, identifying it as a novel mutation. The study identified a novel FBN1 mutation, c.8226+5del, that may be associated with clinical features such as low-set ears and distinctive facial characteristics in the proband. This mutation likely affects normal mRNA splicing, altering the structure and function of Exon 64 and potentially contributing to the development of autosomal dominant MFS.

Evaluation of the Importance of Genetic Polymorphisms in Genes Expressing Cancer-Metabolizing Enzymes (Cyp1a1 and Gstm1) in Oral Submucous Fibrosis.

Genetic polymorphisms are common and contribute significantly to human illnesses. This study was carried out to evaluate the importance of genetic variations in the genes expressing cancer-metabolizing enzymes (CYP1A1 and GSTM1) in individuals experiencing oral submucous fibrosis (OSMF). Based on the clinical and histological characteristics of OSMF, 40 patients were chosen for the study; 10 of these patients had considerable polymorphism and malignant transformation; therefore, they were placed in a different group. After receiving written agreement, 30 normal subject patients were also picked for the study. For both normal and OSMF patients, tissue samples and 2 ml of peripheral venous blood were drawn from the arm vein and placed in a heparinized test tube. Electrophoresis on 0.8% agarose gel was used to verify genomic DNA. The GSTM1 polymorphism, CYP1A1 polymorphism was 10.41% and 15.27% in normal subjects. The GSTM1 polymorphsm, CYP1A1 polymorphism was 16.21% and 8.14% in early OSMF. The GSTM1 polymorphsm, CYP1A1 polymorphism was 20.31% and 28.56% in moderate OSMF. The GSTM1 polymorphsm, CYP1A1 polymorphism was 8.12% and 12.54% in moderate OSMF. Finally, GSTM1 polymorphism was 60.47% in OSMF+ CA, while CYP1A1 polymorphism was 40.21%. The GSTM1 gene polymorphism and CYP1A1 gene polymorphism were maximum in OSMF+ CA category. There is a significant role of genetic variations in the genes expressing cancer-metabolizing enzymes (CYP1A1) and GSTM1 in individuals experiencing OSMF.

Transcriptomic Profiling in Low-Risk Thyroid Cancer Induced by Microwave Ablation.

Peripheral blood mononuclear cells (PBMCs) serve as the immune system's primary transportation hub outside of the affected ablated tissue. This study aims to explore the transcriptomic profiling of the immune response in PBMCs induced by microwave ablation (MWA) in low-risk thyroid cancer. For eight patients diagnosed with low-risk thyroid cancer, 10 ml of peripheral venous blood was collected before MWA as well as one day and one month after MWA. mRNA was extracted from PBMCs for transcriptome next-generation gene sequencing and qRT-PCR analyses. The plasma samples were used for chemokine detection purposes. One day and one month after MWA, there were significant changes in GSEA, particularly in the NF-kappa B-TNFα pathway, inflammatory response, and early and late estrogen response. Common changes in differently expressed genes resulted in a significant downregulation of tumor-promoting genes (BCL3, NR6A1, and PFKFB3). One day after low-risk thyroid cancer MWA, GO enrichment analysis mainly revealed processes related to oxygen transport and other pathways. One month after MWA, GO enrichment analysis mainly revealed regulation of toll-like receptor signaling and other pathways. Furthermore, inflammation-related cytokines and regulatory genes, as well as tumor-promoting cytokines and regulatory genes, were downregulated after MWA. This study presents a comprehensive profile of the systemic immune response induced by thermal ablation for treating low-risk thyroid cancer. More significantly, this study provides valuable insight into potential references for systemic antitumor immunity of ablation against low-risk thyroid cancer. This trial is registered with ChiCTR1900024544.

Association between rs1799724 of TNF- α gene and early onset preeclampsia in Chinese: A pilot study

ObjectiveTo investigate the association between polymorphisms of TNF- α (rs1799724, rs1800629), VEGF (rs3025039) and VEGFR1 (rs 722503) and early onset preeclampsia (EOPE) in Chinese. MethodsA total of 132 EOPE patients from January 2016 to December 2018 at the Second Hospital of Tianjin Medical University were selected as the EOPE group, and 156 normal pregnant patients as the Control group. In both groups, 5 ml of peripheral venous blood was obtained after admission. The characteristics of genotype and allele distribution at the four SNPs in the study subjects were examined by matrix-assisted laser desorption ionization time-of-flight mass spectrometric genotyping. ResultsThe genotype frequency distribution and allele frequency distribution of rs1799724 were significantly different between the EOPE group and the Control group (P = 0.002，P = 0.003). The T allele was statistically associated with the development of EOPE under a dominant genetic model (P = 0.001). The genotype and allele frequency distributions of rs1800629, rs3025039, and rs 722503 did not differ significantly between the EOPE group and the Control group (P > 0.05). There was no linkage disequilibrium among rs1799724, rs1800629 and rs3025039 loci, the corresponding haploid cannot be formed. ConclusionsThe rs1799724 of TNF- α gene is a genetic susceptibility locus for EOPE and may be a potential predictors of preeclampsia.

Sanger Sequencing of Exon 14 of RB1 Gene Among North Indian Children with Retinoblastoma

Introduction: The RB1 gene has been studied for decades. A large number of genetic analytical studies on this gene are published in the Western world, but very few studies from India are available online. RB1 is the most critical gene involved in the causation of retinoblastoma, and Exon 14 is one of the highly mutated exons of this gene. Aim: To discover the genetic variants in Exon 14 of the RB1 gene in North Indian retinoblastoma patients. Materials and Methods: This was an inter- and intra-institutional cross-sectional observational study. The Department of Anatomy, Ophthalmology, and Paediatrics at King George’s Medical University UP, Lucknow, collaborated with the Department of Biochemistry, ERA University, Lucknow. After obtaining written informed consent from their parents or legal guardians, 40 clinically and radiologically diagnosed retinoblastoma patients were included as study participants. The age range was 12-84 months. 2 ml of peripheral venous blood was withdrawn from 29 patients, and we obtained tumour tissue from 14 patients. We isolated the genomic DNA, which was amplified by PCR, and the amplified products were sent for Sanger sequencing. Chromatograms were analyzed using the BioEdit software. Results: Most patients (92.5%) were below five years of age. Male preponderance was seen, as there were 37.5% females and 62.5% males. In the majority of cases (72.5%), the tumor was seen unilaterally with predominant involvement of the right eye; while in 27.5% of cases, both eyes were seen affected. We did not observe any variations in the sequences of exon 14. Conclusions: As the RB1 gene has 27 exons, therefore screening for mutations in all exons is required to reach a concrete conclusion. Genetic variants might be located in the intronic regions, which may affect the proper functioning of the RB1 gene; therefore, sequencing of the entire gene will be more helpful. Next Generation Sequencing (NGS) is preferred because of the large size of the gene. KEYWORDS: Retinoblastoma, DNA, PCR, RB1 gene, sequencing.

Features of genetic mutations in children with high myopia combined with peripheral retinal degenerations

The study investigates the features of genetic mutations in children with high myopia combined with peripheral retinal degenerations. Study group was formed from the database of genetic studies of the Scientific and Clinical Center OOO Oftalmic, which consists of 4362 patients referred for medical genetic counseling and molecular genetic testing from 2016 to 2021. Selection criteria were: male and female patients, aged 5-18 years old, who had the following clinical signs: high myopia (>6.00 D) and the presence of peripheral retinal degenerations (PRD). The study considered both isolated cases of ophthalmic pathology, as well as its syndromic forms. The final selection included 40 children. All patients had consulted with a geneticist. Whole-exome sequencing (WES), next generation sequencing (NGS), and single gene sequencing were conducted by taking 5 mL of peripheral venous blood and extracting deoxyribonucleic acid (DNA). In patients with isolated cases of ophthalmic pathology (peripheral retinal degenerations and high myopia) with a confirmed genetic diagnosis, mutations in the COL2A1 gene were detected in 77.4% of cases, and in the COL11A1 gene - in 22.6% of cases. In Stickler syndrome with a confirmed genetic diagnosis, mutations in the COL2A1 gene were detected in 33.3% of cases. In Marshall syndrome, the mutation in the COL11A1 gene was detected in 11.1% of cases. In children with Ehlers-Danlos, Knobloch type 1, Cohen, Marfan, Wagner syndromes mutations in the genes COL5A1, COL18A1, VPS13B, FBN1, VCAN were detected in 55.6% of cases. In 33.3% of cases of Knobloch type 1, Cohen, Wagner syndromes the mutation is found in both copies of the gene (i.e., in both chromosomes), which leads to the development of peripheral retinal degenerations with high myopia. The results of the conducted molecular genetic testing expand our understanding of the mutation spectrum in the genes of children with both isolated cases of ophthalmic pathology, as well as syndromic pathology.

Relationship between BDNF and oxytocin

Converging, albeit scattered data mainly gathered in animals indicate that the neurotrophin brain-derived neurotrophic factor (BDNF) and the nonapeptide oxytocin (OT) interact in a cooperative way.Data in humans are really limited and indirect. Therefore, the aim of the present study was to explore the possible existence of a link between OT and BDNF in humans, by means of two peripheral markers, the platelet-poor-plasmatic-BDNF (PPP-BDNF) and the platelet BDNF (PLT-BDNF) and OT levels.Twenty-six young healthy controls of both sexes who volunteered for the study were included in the study. Fifty ml of peripheral venous blood were drawn from one-night fasting subjects between 8.00 and 9.00 a.m. The BDNF and OT assays were carried out according to common methods. Comparisons for continuous variables were performed by the Student's t-test for variables that follow a normal distribution, and by the Wilcoxon-Mann-Whitney test for variables not normally distributed. The correlations between biological markers were explored by calculating the Pearson's correlation coefficient or Spearman's rank correlation.The results showed that PLT-BDNF (pg/mg proteins, mean ± SD) and PPP-BDNF (pg/ml, mean ± SD) were 1546 ± 1844 and 10111 ± 1892, respectively. The OT levels (pg/ml, mean ± SD) were 13.92 ± 4.54. The OT levels were significantly higher in women than in men. The Spearman's analysis revealed a statistically significant and negative correlation between OT levels and PLT-BDNF (R = −0.543, p = 0.004).The findings of this study highlight the presence of a significant and negative correlation between OT and PLT-BDNF in a small group of healthy controls of both sexes. In any case, despite all the limits of peripheral biomarkers, they suggest that this reciprocal influence might have a downstream homeostatic function dampening one activity when the other is activated or no longer necessary, maybe at the level of the stress and/or immune systems.

Structural insights of novel mutational frames in Bromodomain Containing-2 gene (BRD2) in juvenile myoclonic epilepsy: bed, bench, and laptop profiles

PurposeJuvenile myoclonic epilepsy (JME) is an adolescent onset type of idiopathic generalized epilepsies. Bromodomain containing protein-2 gene (BRD2), a transcriptional regulatory protein, has a susceptible role in the expression of JME. Considering the polymorphic variations observed in exon 3 of the BRD2 gene, we evaluated the molecular interactions with anti-seizure medication in individuals diagnosed with JME. MethodsThe genomic DNA was extracted from 5 mL of peripheral venous blood of JME participants (n = 55) and healthy control subjects (n = 55). Detailed anti-seizure medication and outcomes were noted during the study period. Identified novel mutations at nucleotide and protein sequences, compared by multiple sequence alignment. Wild-type (WT) and mutated-type (MT) structures were investigated for molecular docking and interactions with anti-seizure drugs. ResultsA common variant at c.1707G>A was found among 23 participants, while a single variant at c.1663ins C was found in one participant. The deletion positions were observed at c.1890delA, c.1892A>T, c.1895A>T, c.1896G>T, c.1897T>C, c.1898T>C, c.1899C>T, c.1900G>T, c.1901C>T and c.1902A>T exhibiting stop codon after p.111Pro>stop; these variants resulted in a truncated protein. In silico analysis was conducted to validate changes; docking analysis showed that novel variant has a significant role in the interactions with anti-seizure drugs. SignificanceBesides clinical and genetic outcomes, ∼5.45% unique genetical variations were observed in the participants. Significant mimicked at the binding site position (92–111) of human BRD2 ranges ∼8.2%, ∼16.4%, and ∼10.6%. Further, research is needed to identify the importance of polymorphism alterations at the binding site and their molecular interactions with anti-seizure drugs, which might be confirmed in a diverse population with JME.

Efficacy relevance of PD-L1 expression on circulating tumor cells in metastatic breast cancer patients treated with anti-PD-1 immunotherapy.

Breast cancer has become the leading cause of cancer mortality in women. Although immune checkpoint inhibitors targeting programmed death-1 (PD-1) are promising, it remains unclear whether PD-L1 expression on circulating tumor cells (CTCs) has predictive and prognostic values in predicting and stratifying metastatic breast cancer (MBC) patients who can benefit from anti-PD-1 immunotherapy. Twenty six MBC patients that received anti-PD-1 immunotherapy were enrolled in this study. The peptide-basedPep@MNPs method was used to isolate and enumerate CTCs from 2.0ml of peripheral venous blood. The expression of PD-L1 on CTCs was evaluated by an established immunoscoring system categorizing into four classes (negative, low, medium, and high). Our data showed that 92.3% (24/26) of patients had CTCs, 83.3% (20/26) of patients had PD-L1-positive CTCs, and 65.4% (17/26) of patients had PD-L1-high CTCs. We revealed that the clinical benefit rate (CBR) of patients with a cut-off value of ≥ 35% PD-L1-high CTCs (66.6%) was higher than the others (29.4%). We indicated that PD-L1 expression on CTCs from MBC patients treated with anti-PD-1 monotherapy was dynamic. We demonstrated that MBC patients with a cut-off value of ≥ 35% PD-L1-high CTCs had longer PFS (P = 0.033) and OS (P = 0.00058) compared with patients with a cut-off value of < 35% PD-L1-high CTCs. Our findings suggested that PD-L1 expression on CTCs could predict the therapeutic response and clinical outcomes, providing a valuable predictive and prognostic biomarker for patients treated with anti-PD-1 immunotherapy.

Abstract P5-02-12: Prognostic relevance of PD-L1 expression on circulating tumor cells in metastatic breast cancer patients treated with anti-PD-1 immunotherapy

Abstract Rationale: Breast cancer has become the leading cause of cancer mortality in women. Although immune checkpoint inhibitors targeting programmed death-1 (PD-1) are promising, it remains unclear whether PD-L1 expression on circulating tumor cells (CTCs) has predictive and prognostic values in predict and stratify metastatic breast cancer (MBC) patients who can benefit from anti-PD-1 immunotherapy. Methods: Twenty six MBC patients that received anti-PD-1 immunotherapy were enrolled in this study. The peptide-based Pep@MNPs method was used to isolate and enumerate CTCs from 2.0 ml of peripheral venous blood. The expression of PD-L1 on CTCs was evaluated by an established immunoscoring system categorizing into four classes (negative, low, medium, and high). Results: Our data showed that 92.3% (24/26) of patients had CTCs, 83.3% (20/26) of patients had PD-L1-positive CTCs, and 65.4% (17/26) of patients had PD-L1-high CTCs. We revealed that the clinical benefit rate (CBR) of patients with a cut-off value of ≥ 35% PD-L1-high CTCs (66.6%) was higher than the others (29.4%). We indicated that PD-L1 expression on CTCs from MBC patients treated with anti-PD-1 monotherapy was dynamic. We demonstrated that MBC patients with a cut-off value of ≥ 35% PD-L1-high CTCs had longer PFS (P&amp;lt; 0.05) and OS (P&amp;lt; 0.01) compared with patients with a cut-off value of &amp;lt; 35% PD-L1-high CTCs. Conclusion: Our findings suggested that PD-L1 expression on CTCs could predict the therapeutic response and clinical outcomes, providing a valuable predictive and prognostic biomarker for patients treated with anti-PD-1 immunotherapy. Citation Format: Ying Zhou, Jinmei Zhou, Haoyuan Shi, Zhiyuan Hu, Yanlian Yang, Zefei Jiang, Tao Wang. Prognostic relevance of PD-L1 expression on circulating tumor cells in metastatic breast cancer patients treated with anti-PD-1 immunotherapy [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P5-02-12.

Genetic analysis of neurodevelopmental disorders in children

PurposeTo explore the genetic cause of children with unidentified etiology of neurodevelopmental disorders, thus providing references for the diagnosis, treatment and genetic counseling.Design and methodsChildren with neurodevelopmental disorders but unidentified etiology in the Child Healthcare Department, Children's Hospital of Nanjing Medical University from November 2018 to December 2021 were retrospectively analyzed. A total of 2 ml of peripheral venous blood was collected from the child and their parents for the whole exome sequencing (WES) and copy number variation (CNV) detection. Male children were subjected to fragile X syndrome testing to determine the genetic mutations. For those with positive results, Sanger sequencing was performed to explore the mutations in the gene sites and pedigrees.ResultsA total of 488 (33.5%) pathogenic variations were detected among 1,457 global developmental/intellectual disabilities (GDD/ID) children, including 362 (24.9%) cases of monogenic mutations, and 111 (7.6%) cases of chromosomal microdeletions or microduplications. There were 15/780 (1.92%) male children with fragile X syndrome. Single point mutations were detected in 277/362 (76.5%) and 85/362 (23.5%) male and female GDD/ID children, respectively, including 295 (81.5%) cases of missense mutations, 32 (8.8%) cases of frameshift mutations, 5 (2.2%) cases of non-sense mutations and 30 (8.3%) cases of splice site mutations. In addition, there were 166 (45.8%) cases of autosomal inheritance and 196 (54.2%) cases of X-linked inheritance. The X chromosome abnormalities were mostly observed in 362 GDD/ID children with monogenic mutations, including 15 cases of the AFF2 gene mutation, 13 cases of the MECP2 gene mutation and 12 cases of the HUWEI gene mutation. The CREBBP gene mutation was the most common autosome abnormality in GDD/ID children with monogenic mutations, which was detected in five cases. There were 74 cases of chromosomal microdeletions, 31 cases of chromosomal microduplications and six cases of both. A total of 114 novel pathogenic mutations responsible for GDD/ID were found, including four novel mutations in MECP2 and TRAPPC9 genes.ConclusionThe incidence of genetic abnormalities remains high in NDD children. Abundant novel mutations are responsible for GDD/ID in children, and can be used as references in the diagnosis of neurogenetic diseases.

Clinical significance of circulating tumor cells testing in auxiliary diagnosis ofgastrointestinal cancer

Routine diagnostic tools and serum biomarkers of gastrointestinal (GI) cancer has limits in detecting early and micrometastasis,and circulating tumor cells (CTCs) had emerged as a promising metrics to complement this gap. The presentstudy is designed to explore technical feasibility of using CTCs as an auxiliary diagnostic tool in GI cancer. Over all 70inpatients with GI cancer and 30 healthy volunteers were recruited, and 10 mL of peripheral venous blood was collectedfrom all subjects. CTCs were detected by microfluidic blood rare cell analysis technique, and the sensitivity and specificityof CTCs in GI cancer diagnosis were derived from comparison with the pathological diagnosis results and tumor serummarker results. Compared with the healthy volunteers, the CTCs levels of the patients in gastrointestinal cancer group weresignificantly increased. Advanced stage subjects demonstrated higher level of CTCs, yet without statistical significance. Thesensitivity of CTCs to diagnose stage I to IV disease were 84.62%, 94.12%, 94.44%, and 100.00% respectively, yieldingcomprehensive sensitivity was 92.56% and specificity was 89.66%. Combined detection of CTCs and four tumor serummarkers was helpful in detecting positive rate, but without statistical signifiance compared with detecting CTCs alone. Ourstudy demonstrates the value of CTCs as an auxiliary diagnostic method for gastrointestinal cancer, and is expected to makeup for the deficiency of routine tissue biopsy, which can be used alone or in combination with conventional serum tumormarkers that can greatly promote the clinical diagnosis/prognosis of gastrointestinal cancer

The Role of Exosome MicroRNA-103 in the Proliferation and Invasion of Hepatoma Cells and Its Mechanism

Liver cancer (HCC) is a common malignant tumor whose incidence is increasing worldwide, but existing chemotherapeutic agents are not ideally effective drugs and have considerable resistance to chemotherapy. Exosome microRNA-103 plays an important role in the proliferation and invasion of liver cancer cells. The purpose of this article is to investigate the role and mechanism of exosome microRNA-103 in hepatocellular carcinoma cell proliferation and invasion. 84 patients with hepatocellular carcinoma diagnosed in a hospital from June 2017 to June 2020 were selected. The average age was 60.13±6.99 years. When the patient was fasting, 3 mL of peripheral venous blood was taken. And 50 healthy control groups were established, with an average age of 59.98±8.18 years old. 3 ml of peripheral venous blood was collected on an empty stomach to compare the cell proliferation rate and invasion rate. The results of the study showed that the number of stage III hepatocellular carcinoma cells invaded at 6h was 68.9, and it changed to 89.4 at 12h, and 106.4 at 24h; compared with that, the cell invasion rate in stage IV was higher. The number of stage IV hepatocellular carcinoma invasions at 6h was 68, which changed to 94.5 at 12h and 112.4 at 24h.

Prognostic Relevance of Estrogen Receptor Status in Circulating Tumor Cells in Breast Cancer Patients Treated With Endocrine Therapy.

Recently, female breast cancer (BC) has surpassed lung cancer to occupy the first place of the most commonly diagnosed cancer. The unsatisfactory prognosis of endocrine therapy for breast cancer might be attributed to the discordance in estrogen receptor (ER) status between primary tumors and corresponding metastases, as well as temporal and spatial receptor status heterogeneity at point-in-time between biopsy and treatment. The purpose of this study was to evaluate the prognostic and predictive value of ER status in circulating tumor cells (CTCs) in BC patients. We analyzed ER expression on CTCs isolated using the Pep@MNPs method in 2.0 ml of blood samples from 70 patients with BC and 67 female controls. The predictive and prognostic value of ER expression in CTCs and immunohistochemistry results of biopsies for progression-free survival (PFS) and overall survival (OS) of patients in response to therapies were assessed. The detection rate for CTCs was 95.71% (67/70 patients), with a median of 8 CTCs within 2 ml of peripheral venous blood (PVB). A concordance of 76.56% in ER status between CTCs and corresponding primary tumor and 69.23% between CTCs and corresponding metastases was observed. We also found that patients with ER-positive CTCs (CTC ER+) had longer PFS and OS than those without ER-positive CTCs (CTC ER-). Our findings suggested that ER status in CTCs of BC patients may provide valuable predictive and prognostic insights into endocrine therapies, although further evaluation in larger prospective trials is required.

Dynamic Changes and Clinical Significance of White Blood Cells and Platelets in Patients with Traumatic Brain Injury

Traumatic brain injury (TBI) is becoming the main cause of death threatening human life because of its high disability rate, high mortality rate and great social harm. The purpose of this study was to investigate the dynamic changes and clinical significance of leukocytes (WBC) and platelets (PLT) in patients with traumatic brain injury. In the process of specimen collection, 4ml of peripheral venous blood was collected 24 hours, 4 days, 7 days, 14 days and 21 days after craniocerebral injury using sterile, non-heat source and non-endotoxin test tube. 2ml venous blood was added into EDTA anticoagulant blood collection vessel to detect the changes of peripheral blood leukocytes and platelets in patients with craniocerebral injury. In addition, 2 ml venous blood was added into the procoagulant blood collection vessel, and the serum was separated and frozen for the determination of CRP and TSP1. All patients were evaluated with Glasgow outcome score at discharge. When detecting PLT and WBC, Sysmex K4500 hematology analyzer was used to routinely measure the number of PLT and WBC in peripheral blood at different time points. In the process of CRP (C-reactive protein) measurement, adjust the pipette to 60 pi, and add 50 pi of developer a and developer B into the micropore respectively. Then it vibrates on a mini vibrator for 5 seconds to ensure mixing. Preheat the microplate reader for 30 minutes, place the microplate in the microplate reader, set the parameters and read the values. When measuring TSP1 (thrombin sensitive protein 1), adjust the micropipette to 50 pi, then add 50pl termination solution into the micropore respectively, and then vibrate on the micro vibrator for 30 seconds to ensure full mixing. Further analysis showed that there was no difference between the non-infection group and the normal control group on the 4th day (P &gt; 0.05). This study is helpful to the timely treatment of patients with traumatic brain injury.

Dynamic Changes and Clinical Significance of White Blood Cells and Platelets in Patients with Traumatic Brain Injury

Traumatic brain injury (TBI) is becoming the main cause of death threatening human life because of its high disability rate, high mortality rate and great social harm. The purpose of this study was to investigate the dynamic changes and clinical significance of leukocytes (WBC) and platelets (PLT) in patients with traumatic brain injury. In the process of specimen collection, 4ml of peripheral venous blood was collected 24 hours, 4 days, 7 days, 14 days and 21 days after craniocerebral injury using sterile, non-heat source and non-endotoxin test tube. 2ml venous blood was added into EDTA anticoagulant blood collection vessel to detect the changes of peripheral blood leukocytes and platelets in patients with craniocerebral injury. In addition, 2 ml venous blood was added into the procoagulant blood collection vessel, and the serum was separated and frozen for the determination of CRP and TSP1. All patients were evaluated with Glasgow outcome score at discharge. When detecting PLT and WBC, Sysmex K4500 hematology analyzer was used to routinely measure the number of PLT and WBC in peripheral blood at different time points. In the process of CRP (C- reactive protein) measurement, adjust the pipette to 60 μ l, and add 50 μl of developer a and developer B into the micropore respectively. Then it vibrates on a mini vibrator for 5 seconds to ensure mixing. Preheat the microplate reader for 30 minutes, place the microplate in the microplate reader, set the parameters and read the values. When measuring TSP1 (thrombin sensitive protein 1), adjust the micro pipette to 50 μ l, then add 50μl termination solution into the micropore respectively, and then vibrate on the micro vibrator for 30 seconds to ensure full mixing. Further analysis showed that there was no difference between the non-infection group and the normal control group on the 4th day (P &gt; 0.05). This study is helpful to the timely treatment of patients with traumatic brain injury.

Evaluation of serum alanine aminotransferase and aspartate aminotransferase enzyme levels in women patients with chronic periodontitis

ABSTRACT The authors’ aim in this study was to investigate the relationship between chronic periodontitis and liver status using alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in women patients. The researchers conducted a case-control study on women patients referred to Dental School. The researchers collected 5 ml of peripheral venous blood from women for the laboratory process after performing periodontal examination. Participants were 124 women aged 25–50 years (62 cases and 62 control). The difference in serum levels of ALT and the difference in serum levels of AST between the two groups were not statistically significant. However, there was a significant correlation between liver enzymes and periodontitis parameters.

Evaluation of genetic polymorphism of interleukin-1β (rs16944) in chronic and aggressive periodontitis in a group of the Bengali population of West Bengal, India

Objectives: The genetic basis of periodontitis was demonstrated by formal genetic studies which were focused on a range of various candidate genes selected for their roles in the immune system like genes of the interleukins (ILs) which regulate the intensity of host immunity-inflammatory response. This regulation of host response may be associated with the genetic polymorphisms, specific single-nucleotide polymorphisms of the genes of various ILs. Interleukin-1 (IL-1) is a principal mediator of inflammatory responses acting on many cell types and is itself produced by many different cells, including macrophages, endothelial cells, B cells, fibroblasts, epithelial cells, astrocytes, and osteoblasts in response to microorganisms, bacterial toxins, and complement components. In the present study, an attempt has been made to explore the role of IL-1β-511 (rs16944) genetic polymorphism in chronic as well as aggressive periodontitis in a group of the Bengali population of West Bengal, India. Materials and Methods: A total of 88 Bengali patients of both sexes were recruited in this study and they were divided into three groups: Group A (chronic periodontitis group), Group B (aggressive periodontitis group), and Group C (healthy control). The clinical parameters taken into consideration in the assessment of chronic and aggressive periodontitis were plaque index, calculus index, gingival index, probing pocket depth (PPD), and clinical attachment loss (CAL). A 3 ml of peripheral venous blood was collected from each selected participant and transferred to 3% EDTA containing serum vial and stored at −20°C for DNA extraction. DNA extraction was performed by the phenol chloroform method and ethanol precipitation. Genotyping of extracting DNA samples was carried out for locus IL-1β-511 (rs16944) by real-time polymerase chain reaction. Hardy–Weinberg equilibrium was tested for the gene polymorphism and association between genotypes and cases was examined by odds ratio with 95% confidence interval (CI) and Chi-square analysis using R programming software. Allelic frequencies were calculated according to the number of different alleles observed and the total number of alleles examined. Statistical significance was defined as P &lt; 0.05. Results: On analysis of minor allele frequencies of total periodontitis cases and control, the results were found to be statistically insignificant with P = 0.9799. Minor allele frequency of overall periodontitis cases and controls was 0.405 and 0.403, respectively; the odds ratio was 1.008 and 95% CI ranges from 0.53 to 1.88. Conclusion: The present study suggested no association of single-nucleotide polymorphism of IL-1β-511 (rs16944) with total periodontitis cases (both chronic periodontitis and aggressive periodontitis) in the present study cohort.

Nonsyndromic deafness due to compound heterozygous mutation of the CDH23 gene

Objective:To identify the pathogenic gene mutation of two patients with non-syndromic deafness（NSHL）. Methods:Two patient with NSHL and their parents were selected in the research object. Each participant provided 3-5 mL of peripheral venous blood, which was used to establish a DNA library. Next generation sequencing was used to detect the sequence of the patient's genome, and the sequencing results were compared with the human genome sequence （GRCh）37/hg19. Sanger sequencing was used to verify the parents' genome sequence. Finally the patient's pathogenic gene mutation was confirmed.Amino acid conservatism and single nucleotide polymorphisms of the mutant sites were analyzed using a variety of databases and software. Results:The mutation was located to CDH23 gene in the chromosomal location 10q21-q22. Complex heterozygous mutations consist of c. 1343T>C and c. 7991_7993delTCA. Parents are heterozygous carriers of a single mutation. Conclusion:The next generation sequencing technology were used to screen the pathogenic gene mutation of inherited deafness. Combined with the genetic sequencing results of parents, the specific pathogenic gene mutation of deafness patients can be identified. While the pathogenicity of complex heterozygous mutation were explained by various pathogenicity analysis methods.