Beta Gene Research Articles

TCR Repertoire Analysis During Therapeutic Interventions in Liver Diseases Using Next-Generation Sequencing.

The T cell receptor (TCR) can recognize a vast number of antigens and is closely associated with the pathogenesis of various diseases including autoimmune diseases and malignancies. However, the clinical significance of the TCR repertoire and its post-treatment changes remain unclear in liver diseases. We performed next-generation sequencing (NGS)-based TCR analysis using DNA obtained from peripheral blood mononuclear cells (PBMCs) of healthy donors (HD, n = 5), primary biliary cholangitis (PBC, n = 5), autoimmune hepatitis (AIH, n = 5), and hepatocellular carcinoma (HCC, n = 5) and evaluated the changes after treatment. Baseline TCR repertoire analysis demonstrated that TCR clonotype usage is restricted and diversity is low in all three disease groups (PBC, AIH, and HCC), particularly in PBC and AIH compared to HD (p < 0.05). Following treatment, clonotype usage and diversity did not change significantly, except in AIH, where diversity decreased further (p < 0.05 for clone Shannon diversity and clone evenness). Disease-specific usage of TCR beta genes and specific changes after therapy were observed in all groups. Analysis of clonotypes shared with other individuals (public clonotypes) revealed that nine public clonotypes in PBC, eight in AIH, and eight in HCC disappeared after treatment. Motif analysis identified one characteristic motif (NQPQH) in PBC. The diversity of the TCR repertoire, TCR beta chain usage, clonotypes, and motifs and their post-treatment changes are disease-specific in each liver disease, indicating that further TCR repertoire studies are needed to accelerate the understanding of liver disease pathogenesis from an immunological perspective.

Assessment of liver x receptor messenger RNA beta and microRNA-146a in a group of Egyptian patients with Behçet’s disease

Abstract Background and objectives Behçet’s disease (BD) is an ongoing systemic vasculitis with mouth and genital ulceration and eye, skin, and systemic affection. It has considerable morbidity and mortality, and a high incidence and a higher male-to-female affection characterize Egypt. We aimed to evaluate liver x receptor messenger RNA beta (LXR β) and microRNA-146a (miR-146a) gene expression in Behçet’s disease cases in Egypt to relate them with clinicopathological features. Subject and method Eighty Egyptian individuals were split into two groups for the current study: Forty Behçet’s disease cases and forty healthy controls matched by age and gender from the Rheumatology Department at Helwan University Hospital, Egypt. The transformed score, also known as the Behçet’s Disease Current Activity Form (BDCAF), was utilized to measure disease activity. To assess the amounts of LXR β and miR-146a serum expression via real-time PCR, a blood sample was obtained. Results There was a downregulation of both LXR β and miR-146a levels that significantly differed between the BD group and the control group (p = 0.0001 and 0.0001, respectively). There is a noteworthy inverse relationship between the expression level of LXR β and BDCAF Patients Index (r = − 0.79 and p = 0.0001) was found. Regarding miR-146a, it had a reverse correlation with BDCAF Patients Index (r = − 0.89 and p = 0.0001). Conclusion LXR β and miR-146a were found to be significant non-invasive predictor biomarkers for Behçet’s disease and can indicate disease activity.

Mycosis Fungoides: Histopathology and T Cell Receptor Gene Rearrangements in Indian Patients

Introduction: Mycosis fungoides (MF) is the commonest cutaneous T cell lymphoma. MF presents in three stages the patch, plaque, and tumor. The distinction of patch-stage MF from inflammatory dermatoses is a diagnostic challenge. Need for the Study: MF shows T cell receptor gamma and beta gene rearrangements (TCRG and TCRB), detectable by polymerase chain reaction (PCR). Data about the clonality of infiltrates in our population and their utility in diagnosis is lacking. Materials and Methods: Skin biopsies from 50 patients with mycosis fungoides from January 2007 to July 2015 were studied for microscopic features. TCR gamma and beta gene rearrangement assays were performed on 14 patients where formalin fixed paraffin embedded (FFPE) was available. DNA was extracted using the phenol-chloroform method. PCR was performed using primers from Invivoscribe Identiclone assay. Clonality was analyzed using Agarose gel electrophoresis. Results: Of the 50 cases, 37 were patch, nine were plaque, and four were tumor stage. All cases showed disproportionate lymphocyte epidermotropism, 38 showed lymphocyte atypia, and 33 showed adnexotropism (11 eccrotropism and 22 folliculotropism). Clonality was assessed in 14 cases. Of these, 3/9 patch stages were monoclonal (1/3 monoclonal for beta, and 2/3 monoclonal for gamma and beta). 3/3 plaque stage MF was monoclonal (1/3 for gamma, 2/3 for both), and 1/2 tumor stage MF was monoclonal for gamma and beta receptors. Conclusion: The patch stage was the most common presentation. TCR gene rearrangements were noted in 50% of MF, with TCR gamma rearrangements being more common. Plaque and tumor stages yielded higher positive rates.

Joint Analysis of CCAAT/Enhancer-Binding Protein Beta and Interleukin 1 Beta in the Treatment and Prognosis of Diffuse Large B-Cell Lymphoma.

The purpose of this study is to investigate the correlation between elevated levels of CCAAT/enhancer-binding protein beta (CEBPB) gene expression and unfavorable outcomes in diffuse large B-cell lymphoma (DLBCL). The goal is to elucidate potential therapeutic targets associated with this relationship. Differential expression and survival analyses were conducted using data from the Gene Expression Omnibus (GEO) database. The functions of CEBPB in DLBCL cells were investigated through cell culture, RNA extraction, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot. In addition, a weighted gene co-expression network analysis (WGCNA) was performed to pinpoint gene modules associated with CEBPB. Furthermore, experimental validation was carried out to explore the interaction between CEBPB and interleukin 1 beta (IL1B). High levels of CEBPB expression are prominently observed in DLBCL, with its overabundance significantly linked to the diagnosis of DLBCL. Survival analysis reveals that patients exhibiting elevated CEBPB expression tend to experience a poorer prognosis. Further validation confirmed CEBPB's role in promoting DLBCL cell proliferation and cell cycle progression. WGCNA identified CEBPB-related gene modules, with IL1B identified as a potential regulatory gene of CEBPB. The presence of high levels of IL1B has been correlated with an unfavorable prognosis in individuals diagnosed with DLBCL. Experiments demonstrate that IL1B promotes DLBCL cell proliferation through CEBPB. This study reveals the significant roles of CEBPB and IL1B in DLBCL, providing new theoretical foundations and potential molecular targets for the treatment and prognosis of DLBCL.

Gamma irradiation green synthesis of (polyacrylamide/chitosan/silver nanoparticles) hydrogel nanocomposites and their using as antifungal against Candida albicans and anti-cancer modulator

Silver nanoparticles-loaded hydrogel nanocomposites are exploited for medicinal and pharmaceutical applications. Hydrogel nanocomposites were prepared from acrylamide (Am), chitosan (CS) and AgNO3 utilizing gamma rays. Diverse variables were applied in preparation of silver nanoparticles-laoded hydrogel nanocomposites of (PAm/CS)-AgNPs such as influence of radiation dose and influnece of CS concentration. Diverse techniques were utilized to characterize hydrogel nanocomposites; Fourier transform infrared spectroscopy (FTIR), thermal gravimetric analysis (TGA), X-ray diffraction (XRD), Transmission electron microscopy (TEM), energy dispersive X-ray (EDX) and scanning electron microscopy (SEM). Results confirmed formation of silver nanoparticles-loaded hydrogel nanocomposites of (PAm/CS)-AgNPs. Antifungal activity of (PAm/CS)-AgNPs hydrogel nanocomposites on viability of C. albicans was esitmated. Results displayed the efficient microbial inhibition activity of treatment against C. albicans compared to control. Furthermore, (PAm/CS)-AgNPs hydrogel nanocomposite against cervical cancer HeLa cell line was investigated. Cytotoxicity of (PAm/CS)-AgNPs hydrogel nanocomposites on prior cancer cell line empolyed to prohibition of cell growth assesssed by MTT test. HeLa cancer cell is treated by (PAm/CS)-AgNPs for 48 h exposed a potential apoptotic activity by noticeable up-regulation of p53 gene expression. Moreover, anticancer activity was investigated by down-regulation of platelet-based growth variable receptor beta (PDGFR-β), Bcl2, Cathepsine, and MMP-2 gene expression. antioxidant activity was investigated and results showed antioxidant activity of (PAm/CS) hydrogel and (PAm/CS)-AgNPs hydrogel nanocomposite are 87.8% and 62.9%, respectively.

PCR GeneScan Analysis of Rearranged Immunoglobulin or T-Cell Receptor Genes for Clonality Diagnostics in Suspect Lymphoproliferations.

Assessment of the presence of clonal lymphoproliferations via polymerase chain reaction (PCR)-based analysis of rearranged immunoglobulin (IG) or T-cell receptor (TR) genes is a valuable method in the diagnosis of suspect lymphoproliferative disorders. Additionally, this methodology can be used for evaluating dissemination of lymphoma cells and for studying the clonal relationship between multiple (different locations) and consecutive (over time) lymphomas. Here we describe an integrated approach to assess clonality via analysis of Ig heavy chain (IGH), Ig kappa (IGK), TCR beta (TRB), and TCR gamma (TRG) gene rearrangements, based on the standardized multiplex PCRs as originally developed by the European BIOMED-2 consortium (currently named EuroClonality). The described protocol covers the pre-analytical phase of DNA isolation (from formalin-fixed paraffin-embedded and fresh tissues, body fluids, peripheral blood, and bone marrow), the analytical phase of PCR GeneScan analysis, and the post-analytical interpretation of the obtained profiles, following established guidelines.

Oral pre-treatment with Citronellol ameliorates Methotrexate-induced nephrotoxicity in Wistar rats via targeting oxidative stress and inflammation

Background Methotrexate (MTX) is a classical folic acid antagonist widely used in the treatment of malignant and non-malignant disorders. However, its clinical application is often restricted by concomitant adverse effects, including renal damage. Numerous studies have highlighted the role of oxidative stress and inflammation in mediating MTX-related nephrotoxicity. Therefore, the current study aimed to explore the possible renoprotective action of Citronellol (CT), a natural compound with prominent antioxidant and anti-inflammatory activities, against nephrotoxicity induced by MTX. Methods To fulfill our objective, 24 adult male Wistar rats were randomly allocated into four groups: control, MTX, 100 mg/kg CT plus MTX and 200 mg/kg CT plus MTX. At the end of the study, the experimental rats were anesthetized, blood samples were collected for biochemical assays, and the kidneys were surgically removed for biochemical and gene expression analyses, after which all rats were sacrificed by exsanguination. Results Compared to the MTX-treated group, our results revealed that pre-supplementation with 100 or 200 mg/kg CT remarkably ameliorated renal damage biomarkers, including serum urea, serum creatinine, and kidney injury molecule-1 (KIM-1). In addition, pre-treatment with 100 or 200 mg/kg CT enhanced the activity of the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx), diminished renal malondialdehyde (MDA) contents, and attenuated inflammation by suppressing renal nuclear factor kappa B (NF-κB) signaling and diminishing tumor necrosis factor-alpha (TNF-α) gene expression. Moreover, pre-treatment with 200 mg/kg CT markedly reduced interleukin-1 beta (IL-1β) gene expression. Conclusion Our findings demonstrate, for the first time, that CT can serve as a new promising agent for mitigating nephrotoxicity induced by MTX through its antioxidant and anti-inflammatory properties.

Breeding Maize Hybrids with Improved Drought Tolerance Using Genetic Transformation

Drought is considered the main agricultural menace, limiting the successful realization of land potential, and thereby reducing crop productivity worldwide. Therefore, breeding maize hybrids with improved drought tolerance via genetic manipulation is necessary. Herein, the multiple bud clumps of elite inbred maize lines, DH4866, Qi319, Y478 and DH9938, widely used in China, were transformed with the Escherichia coli betA gene encoding choline dehydrogenase (EC 1.1.99.1), a key enzyme in the biosynthesis of glycine betaine from choline, using Agrobacterium to generate betA transgenic lines. After 3–4 consecutive generations of self-pollination in these transgenic plants, progenies with a uniform appearance, excellent drought tolerance, and useful agricultural traits were obtained. We evaluated the drought tolerance of T4 progenies derived from these transgenic plants in the field under reduced irrigation. We found that a few lines exhibited much higher drought tolerance than the non-transformed control plants. Transgenic plants accumulated higher levels of glycine betaine and were relatively more tolerant to drought stress than the controls at both the germination and early seedling stages. The grain yield of the transgenic plants was significantly higher than that of the control plants after drought treatment. Drought-tolerant inbred lines were mated and crossed to create hybrids, and the drought tolerance of these transgenic hybrids was found to be enhanced under field conditions compared with those of the non-transgenic (control) plants and two other commercial hybrids in China. High yield and drought tolerance were achieved concurrently. These transgenic inbred lines and hybrids were useful in marginal and submarginal lands in semiarid and arid regions. The betA transgene can improve the viability of crops grown in soils with sufficient or insufficient water.

Interleukin one beta (IL-1B) gene polymorphisms in Iraqi patients with susceptibility to develop hepatitis C infections

Hepatitis C virus (HCV) infection is the main cause of chronic hepatitis, affecting an estimated 150 million people worldwide. Initial exposure to HCV is most often followed by chronic hepatitis, with only a minority of individuals spontaneously clearing the virus. This study aimed to find an associated relationship between HCV susceptibility and Polymorphic gene of IL‐1B targeting a total of 45 individuals: 30 patients with chronic hepatitis C and 15 healthy control from different cities of Iraqi recruited the Hospital for diseases of the digestive system and liver in Baghdad between November 2022 to February 2023. Genomic DNA was extracted and specific promotor sequence was amplified by polymerase chain reaction (PCR) followed by sequencing then measuring serum concentrations of IL-1β by Enzyme-Linked Immunosorbent Assay (ELISA). The results revealed significant difference in serum levels of IL-1B (P≤0.01), from 1.10-+0.09 in Chronic HCV patients in comparison with its level in healthy controls 7.58± 0.28. Molecular results suggested that single nucleotide polymorphisms in the promoter of IL-1B is probably associated with susceptibility to HCV chronic infection according to this sample of Iraqi patients. In conclusion, the results suggested that variation in serum level and polymorphism in IL-1B is probably associated with susceptibility to hepatitis C virus chronic infection.

Anti-Inflammatory Activity and Wound Healing Ability of Coconut Oil Mouthwash on Gingival Fibroblast Cell In Vitro

Coconut oil-pulling therapy is used for maintaining oral health. The procedure has benefits for the prevention of oral disease, including dental caries, oral malodor, bleeding gums. However, virgin coconut oil (VCO) has unsatisfying oily taste. Therefore, coconut oil mouthwash (CoMW) was recently developed. This in vitro study aimed to evaluate the anti-inflammatory activity of coconut oil mouthwash consisting of 60% v/v virgin coconut oil (VCO), 30% v/v propylene glycol (PG), and 10% v/v distilled water on human gingival fibroblast (HGF) cells compared with the activity on murine macrophage (Raw 264.7) cells. The cytotoxicity of CoMW, 0.12% chlorhexidine gluconate (CHX), VCO, and PG was assessed. IC50 concentration of CoMW, CHX, and PG were 1:8 (v/v), 1:32 (v/v), and 1:16 (v/v), respectively. All tested concentrations of VCO had no impact on cell viability. Their anti-inflammatory effects of each IC50 concentration were further studied. Notably, the IC50 concentration of CHX also significantly inhibited nitric oxide production in lipopolysaccharide (LPS)- activated Raw 264.7 cells. Moreover, the IC50 concentrations of CoMW, VCO, PG, and CHX could suppress the interleukin-1 beta (IL-1β), interleukin-6 (IL-6) and cyclooxygenase-2 (COX-2) gene expressions in LPS-activated HGF cells, while also enhancing the cell migration of HGF cells as likely to the effect observed with the IC50 concentration of CHX. Wound healing ability of CoMW was also demonstrated after testing with a scratch assay. These findings indicated promising potential for coconut oil mouthwash as an effective agent in reducing inflammation and facilitating wound healing.

Compromised actin dynamics underlie the orofacial cleft in Baraitser-Winter Cerebrofrontofacial syndrome with a variant in ACTB.

Craniofacial anomalies encompassing the orofacial cleft are associated with > 30% of systemic congenital malformations. Baraitser-Winter Cerebrofrontofacial syndrome (BWCFF) is a rare genetic disorder attributed to variants in the actin beta (ACTB) or actin gamma genes that are correlated with a range of craniofacial abnormalities, including cleft lip and/or palate. The underlying pathological mechanism of BWCFF remains elusive, and it is necessary to investigate the etiology of orofacial clefts in patients with BWCFF. In this study, we identified a missense variant (c.1043C > T: p.S348L) in the ACTB gene of a patient with BWCFF and concomitant cleft lip and palate. Furthermore, we performed functional assessments of this variant using various disease models such as the MDCK cell line and Xenopus laevis. These models revealed a compromised capacity of mutated ACTB to localize to the epithelial junction, consequently affecting the behavior of epithelial cells. Additionally, we discovered that the mutated ACTB exhibited an impaired ability to bind PROFILIN1, a critical factor in actin polymerization. This defective ability may contribute to the molecular etiology of aberrant epithelial cell adhesion and migration, resulting in orofacial cleft formation in BWCFF.

Expression of steroidogenesis pathway genes in cumulus cells from women with diminished ovarian reserve after gonadotropin administration: A case-control study.

Women with diminished ovarian reserve (DOR) respond differently to gonadotropin medications. This study investigates the relationship between effective gene expression in the steroidogenesis pathway and gonadotropin responsiveness in DOR. In this case-control study, cumulus cells were obtained from women with DOR after gonadotropin administration (n = 20) and normal ovarian reserve (n = 20). They were divided into the following groups, oocyte number 3 and oocyte number 3. After RNA extraction and cDNA synthesis, quantitative polymerase chain reaction was performed to assess the expression levels of cytochrome P450 aromatase (CYP19A1), protein kinase A(PKA), and glycogen synthase kinase 3 beta (GSK3B)genes. The women with DOR had statistically significant lower expression of CYP19A1 and PKA genes in their cumulus cells compared to control group (p = 0.04, and p 0.001, respectively). There was also lower expression of the GSK3B gene in DOR compared to control group, but it was not significant. Although the expression of the CYP19A1, PKA, and GSK3B genes was lower in women with 3 oocytes compared to women with more oocytes, this difference was not statistically significant. In conclusion, DOR may be associated with lower expression of CYP19A1 and PKA genes. Also, considering the decrease in the expression of these genes in people with DOR, the expression of these genes can be used as a tool to predict the treatment.

Clinical characteristics and prognosis of early diagnosed Wilson's disease: A large cohort study.

Few studies have focused on the outcomes of Wilson's disease (WD) diagnosed before age of 5 years. This study aimed to summarize the clinical features of early diagnosed WD and analyse treatment outcomes and the risk factors associated with treatment failure. A total of 139 children confirmed with WD before 5 years were enrolled in this study. Only patients with follow-up over 1 year were analysed with Kaplan-Meier survival analysis. The composite outcomes included death, progression to liver failure or acute hepatitis, development of renal or neurological symptoms and persistent elevation of alanine aminotransferase (ALT). The treatment failure was defined as occurrence of at least one of above outcomes. Among 139 WD patients at diagnosis, two (1.4%) WD patients presented with symptomatic liver disease, whereas 137 (98.6%) were phenotypically asymptomatic, including 135 with elevated ALT and 2 with normal liver function. Median serum ceruloplasmin (Cp) was 3.1 mg/dL, and urinary copper excretion was 87.4 μg/24-h. There were 71 variants identified in the the copper-transporting ATPase beta gene, and 29 were loss of function (LOF). 51 patients with LOF variant were younger at diagnosis and had lower Cp than 88 patients without LOF. Among 93 patients with over 1 year of follow-up, 19 (20.4%) received zinc monotherapy, and 74 (79.6%) received a zinc/D-penicillamine combination therapy. 14 (15.1%) patients underwent treatment failure, and its occurrence was associated with poor compliance (p < .01). Cp is a reliable biomarker for early diagnosis, and zinc monotherapy is an effective treatment for WD during early childhood. Good treatment compliance is critical to achieve a favourable outcome.

Nephroprotective Effect of Ferula assa-foetida Oleo Gum Resin on Type 2 Diabetic Rats.

Diabetic nephropathy is one of the main causes of kidney failure in the end stage of diabetes worldwide. On the other hand, asafoetida is a gum whose hypoglycemic effects have been proven. The present study was conducted with the aim of using asafoetida to prevent diabetic nephropathy. Diabetes was induced by a high-fat diet (60%) and streptozotocin injection (35 mg/kg) in rats. Diabetic rats were treated with an oral dose of 50 mg/kg of asafoetida for 8 weeks. At the end of the experiment, serum and urine parameters were examined. Antioxidant enzymes and lipid peroxidation levels in the kidney were also determined along with its histological examination. The expression levels of tumor necrosis factor-alpha and Transforming growth factor beta genes were also evaluated. Glucose, cholesterol, triglyceride, and HbA1c concentrations were significantly reduced in the asafoetida 50. On the other hand, in the treatment group, serum creatinine, urea, and albumin levels decreased and increased in urine. Antioxidant enzymes in the kidney improved significantly, and the expression of tumour necrosis factor-alpha and transforming growth factor-beta genes decreased. Histopathological examination also showed that necrosis, epithelial damage, and leukocyte infiltration increased in the diabetic and decreased in the treatment group. The result of biochemical analysis, enzymatic, and histological examinations showed that asafoetida may delay the progression of diabetic nephropathy due to the presence of anti-inflammatory and antioxidant activities.

Autochthonous probiotic bacteria improve intestinal pathology and histomorphology, expression of immune and growth-related genes and resistance against Vibrio alginolyticus in Asian seabass (Lates calcarifer).

The study isolated two strains of intestinal autochthonous bacteria Lactiplantibacillus plantarum1 (MH155966.1) (L1) and Lactiplantibacillus plantarum2 (MH105076.1) (L2) from the Choobdeh Abadan region. The aim of this study was to investigate the effects of different strains of probiotic bacteria on the growth performance, digestive enzyme activity, histopathologic and histomorphometric characterization of the intestine, expression of immune and growth related genes, and evaluate Lates calcarifer resistance against Vibrio alginolyticus. To achieve this, for each treatment 60 L. calcarifer juveniles (75 ± 12g) were randomly distributed in three fiberglass tanks (300L) and fed for 45 days. The treatments were established as Diet 1 (control diet); L1 (diet with Lb. plantarum isolated 1); L2 (diet with Lb. plantarum isolated 2) with a bacterial concentration of 1 × 109 CFU/g. Nine fish from each treatment were sampled and examined, after euthanasia. The fish were placed 2cm from the beginning of the intestine for microscopic sampling of villi height, villi width and thickness of the epithelium, with 3 treatments: The result showed differences in the mean values of total weight were found at the end of the experiment. After 45 days of culture, the fish fed with L1 had higher (P < 0.05) growth performance than the other treatment groups. But at the end of the trial, in L2, the digestive enzyme activities were higher (P < 0.05) than the other treatment groups. The fishes fed diets supplemented with the L2 group, like the digestive enzyme activities test, presented an increase in the thickness of the epithelium of the intestine, and villus height, and villus width were greatest in L2. Fish feeding with L1 and L2 probiotics induced higher transcription levels of interleukin-10 (IL-10), granulocyte-macrophage colony-forming cells (GMCFC), epidermal growth factor (EGF), and Transforming Growth Factor Beta (TGF-β) genes in the gut, which may correlate with better immune and hematological parameters in these groups. The results of the challenge test revealed that the percentage of survival was significantly higher in L1 (76.2%) and L2 (80.95%) treatments than in the control (P < 0.05). These results indicate that host-derived probiotics (Lb. plantarum) have significant potential as important probiotics to enhance nutrient utilization, Digestive enzymes, and metabolism by increasing the gut surface area of Lates calcarifer juveniles at 45 days of culture.

Glycine Receptor Beta Subunit (GlyR-β) Promotes Potential Angiogenesis and Neurological Regeneration during Early-Stage Recovery after Cerebral Ischemia Stroke/Reperfusion in Mice.

Ischemic stroke is mainly caused by cerebral artery thrombosis. This study investigated the role of glycine receptor beta subunit (GlyR-β) in the recovery from cerebral ischemia stroke/reperfusion. The oxygen glucose deprivation and recovery (OGD/R) bEnd3 cell model and the middle cerebral artery occlusion/reperfusion (MCAO/R) mouse model were used in this study. Expression of both the GlyR-β gene and vascular endothelial growth factor (Vegf), cell proliferation, and tube formation ability was decreased in bEnd3 cells after OGD/R, and was reversed by overexpression of GlyR-β. Neurological function, asindicated by Zea Longa scores, area of cerebral ischemia, and pathological changes were increased in mice after MCAO/R, and were ameliorated by overexpression of the glycine receptor beta (Glrb) gene at 24 h and 7 d after MCAO/R. Expression of GlyR-β and Gap-43 was decreased, and the expression of CD34, Vegf, and Bdnf, and cell growth as determined by a bromodeoxyuridine (BrdU) assay, increased in the affected brain tissue of MCAO/R mice in a time-dependent manner. GlyR-β overexpression resulted in enhanced expression of CD34, Vegf, Growth association protein 43 (Gap-43), and brain-derived neurotrophic factor (Bdnf) and cell growth in affected brain tissue of MCAO/R mice in a time-dependent manner. GlyR-β promoted potential angiogenesis and neurological regeneration in affected brain tissue, thus promoting recovery from cerebral ischemia stroke/reperfusion.

A novel mutation in the ATP7B gene causing hepatolenticular degeneration in a Chinese family: A case report.

Hepatolenticular degeneration (Wilson disease) is an autosomal recessive monogenic disorder caused by mutations in the ATPase copper transporting beta (ATP7B) gene located on human chromosome 13. This gene encodes a copper-transporting P-type ATPase (ATP7B). Recent studies have revealed that the ATP7B gene is predominantly affected by a few hotspot mutations, with the His1069Gln mutation in exon 14 accounting for 50 to 80% of cases. In China, the Arg778Leu mutation in exon 8 is the most prevalent. However, the discovery of novel mutant genes persists. A 56-year-old Chinese female was referred to our hospital with a liver injury and cirrhosis. Her parents, 2 younger brothers, and children exhibited no signs of liver function impairment. Whole-exome sequencing was conducted on the proband's genomic DNA, and Sanger sequencing was performed on 6 family members for first-generation verification. We identified a novel c.3715G > T (p.Val1239Phe) variant mutation in the ATP7B gene in the patient. The ATP7B c.3715G > T (p.Val1239Phe) variant is predicted to impact the copper transport P-type ATPase. When combined with another mutant gene to form a compound heterozygous mutation, it can lead to hepatolenticular degeneration. This discovery broadens the range of pathogenic genes in the ATP7B gene.

Deciphering the dynamic expression network of fiber elongation and the functional role of the GhTUB5 gene for fiber length in cotton based on an introgression population of upland cotton

IntroductionInterspecific introgression between Gossypium hirsutum and G. barbadense allows breeding cotton with outstanding fiber length (FL). However, the dynamic gene regulatory network of FL-related genes has not been characterized, and the functional mechanism through which the hub gene GhTUB5 mediates fiber elongation has yet to be determined. MethodsCoexpression analyses of 277 developing fiber transcriptomes integrated with QTL mapping using 250 introgression lines of different FL phenotypes were conducted to identify genes related to fiber elongation. The function of GhTUB5 was determined by ectopic expression of two TUB5 alleles in Arabidopsis and knockout of GhTUB5 in upland cotton. Yeast two-hybrid, split-luciferase and pull-down assays were conducted to screen for interacting proteins, and upstream genes were identified by yeast one-hybrid, dual-LUC and electrophoretic mobility shift assays. ResultsThe 32,612, 30,837 and 30,277 genes expressed at 5, 10 and 15 days postanthesis (dpa) were grouped into 19 distinct coexpression modules, and 988 genes in the MEblack module were enriched in the cell wall process and exhibited significant associations with FL. A total of 20 FL-QTLs were identified, each explaining 3.34–16.04 % of the phenotypic variance in the FL. Furthermore, several FL-QTLs contained 15 genes that were differentially expressed in the MEblack module including the tubulin beta gene (TUB5). Compared with the wild type, the overexpression of GhTUB5 and GbTUB5 in Arabidopsis suppressed root cell length but promoted cellulose synthesis. Knockout of GhTUB5 resulted in longer fiber lines. Protein-based experiments revealed that GhTUB5 interacts with GhZFP6. Additionally, GhTUB5 was directly activated by GhHD-ZIP7, a homeobox-leucine zipper transcription factor, and its paralogous gene was previously reported to mediate fiber elongation. ConclusionThis study opens a new avenue to dissect functional mechanism of cotton fiber elongation. Our findings provide some molecular details on how GhTUB5 mediates the FL phenotype in cotton.

Mutation Analysis of Exon 1 in the Hemoglobin Subunit Beta (HBB) Gene in Beta-Thalassemia.

Introduction Thalassemia is a widely prevalent monogenic hematological disorder found worldwide. It exists in two forms: alpha- and beta-thalassemia. Alterations in the hemoglobin subunit beta (HBB) gene cause beta-thalassemia, with missense and point mutations affecting beta-globin synthesis. Consequently, genetic screening for beta-thalassemia is essential for genetic counseling, carrier screening, and prenatal diagnosis. Aim and objective This study aims to examine and identify mutations in the exon 1 region of the HBB gene in beta-thalassemia patients from the Vijayapura region. Methods This study involved 47 clinically diagnosed children with beta-thalassemia from a hospital in Vijayapura, India. Detailed clinical histories of all patients were recorded. Genomic DNA was extracted from the blood samples of these patients and subjected to polymerase chain reaction (PCR) using exon-specific primers for the HBB gene. The PCR products were then sequenced using the capillary-based Sanger sequencing method to identify mutations in the HBB gene. Results A total of 47 clinically diagnosed beta-thalassemia patients were included in the study, comprising 30 males and 17 females, aged between one and 20 years. Sequencing analysis of exon 1 in the beta-globin gene identified 17 beta-thalassemia variants. The most common mutation observed was T>G, G>C, C>A, and C>T in the exon 1 region of the HBB gene. Conclusion This study identifies the pattern of beta-thalassemia mutations, aiding in the prevention of the disorder through prenatal diagnosis and genetic counseling. Mutations can alter codon sequences, affecting protein production. Research highlights the importance of a primary prevention program to analyze mutations and sequence variations at the molecular level, thereby helping to address numerous genetic disorders.

Navigating the CRISPR/Cas Landscape for Enhanced Diagnosis and Treatment of Wilson's Disease.

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) system continues to evolve, thereby enabling more precise detection and repair of mutagenesis. The development of CRISPR/Cas-based diagnosis holds promise for high-throughput, cost-effective, and portable nucleic acid screening and genetic disease diagnosis. In addition, advancements in transportation strategies such as adeno-associated virus (AAV), lentiviral vectors, nanoparticles, and virus-like vectors (VLPs) offer synergistic insights for gene therapeutics in vivo. Wilson's disease (WD), a copper metabolism disorder, is primarily caused by mutations in the ATPase copper transporting beta (ATP7B) gene. The condition is associated with the accumulation of copper in the body, leading to irreversible damage to various organs, including the liver, nervous system, kidneys, and eyes. However, the heterogeneous nature and individualized presentation of physical and neurological symptoms in WD patients pose significant challenges to accurate diagnosis. Furthermore, patients must consume copper-chelating medication throughout their lifetime. Herein, we provide a detailed description of WD and review the application of novel CRISPR-based strategies for its diagnosis and treatment, along with the challenges that need to be overcome.