Association between Calf Circumference and Sarcopenia in Kidney Transplant Recipients

Background Sarcopenia may be associated with the long-term prognosis of renal transplant recipients, so it is essential to early and rapidly screening for sarcopenia in these patients. Studies suggest that calf circumference may be an effective indicator in early screening for sarcopenia. Objective To investigate the relationship between calf circumference and sarcopenia, and to explore the feasibility of using calf circumference as a predictive marker for sarcopenia in kidney transplant recipients. Methods We selected 80 kidney transplant recipients who were treated in the department of organ transplantation of Guangdong Second Provincial General Hospital from October 2021 to June 2022 as the subjects. Demographic information (sex, age, educational attainment, marital status) , anthropometric indicators〔height, weight, body mass index (BMI) , calf circumference, mid-arm circumference (MAC) , triceps skin-fold thickness (TSF) , arm muscle circumference (AMC) , waist circumference, hip circumference, waist-to-hip ratio〕, muscle strength indicators (grip strength, pinch strength) , body composition indicators〔appendicular skeletal muscle mass (ASM) , skeletal muscle mass index (SMI) , 50 kHz phase angle (PhA) , body cell mass (BCM) , intracellular and extracellular fluids〕 were collected. Sarcopenia was diagnosed by the Asian Working Group for Sarcopenia 2019 consensus. Kidney transplant recipients who meet the diagnostic criteria were included in the sarcopenia group, and those who do not meet the diagnostic criteria were included in the non-sarcopenia group. Demographic information, anthropometric indicators, muscle strength indicators, and body composition indicators between sarcopenia and non-sarcopenia patient groups were compared by sex. Pearson correlation analysis and Spearman rank correlation analysis were used to observe the correlation of calf circumference with diagnostic indicators of sarcopenia (ASM, SMI, grip strength) . Univariate and multivariate logistic regression analyses were used to explore the influencing factors of sarcopenia. The predictive value of calf circumference for sarcopenia in kidney transplant of male and female subjects was analyzed by using the receiver operating characteristic (ROC) curve, and the area under the curve (AUC) , sensitivity, specificity and optimal cut-off values were calculated. Results A total of 80 patients〔51 men (63.8%) and 29 women (36.2%) 〕 with an average age of (44.3±11.7) years were included, including 19 (23.75%) with sarcopenia, and 61 (76.25%) without. Sex-based analysis found that male patients with sarcopenia group had lower values of weight, BMI, calf circumference, MAC, TSF, hip circumference, grip strength, pinch strength, ASM, SMI, PhA, BCM, extracellular fluid and intracellular fluid than those without (P<0.05) . Female patients with sarcopenia had lower values of weight, BMI, calf circumference, waist circumference, hip circumference, waist-to-hip ratio, ASM, SMI, BCM, extracellular fluid and intracellular fluid than those without (P<0.05) . Correlation analysis showed that the calf circumference had a positive correlation with ASM, SMI and grip strength in both male kidney transplants (rs=0.545, P<0.001; rs=0.540, P<0.001; rs=0.340, P=0.015) and female kidney transplants (rs=0.499, P=0.006; rs=0.578, P=0.001; r=0.426, P=0.021) . Multivariate Logistic regression analysis showed that calf circumference was associated with sarcopenia in kidney transplant〔OR=0.699, 95%CI (0.051, 0.975) , P=0.035〕. ROC analysis revealed that the AUC of calf circumference predicting sarcopenia in kidney transplant of male subjects was 0.799, with the optimal cut-off value, sensitivity and specificity of 33.3 cm, 83.3% and 74.4%, respectively. And the AUC of calf circumference predicting sarcopenia in kidney transplant of female patients was 0.851, with the optimal cut-off value, sensitivity and specificity of 32.2 cm, 100.0% and 59.1%, respectively. Conclusion Calf circumference is associated with sarcopenia and can be used as a predictive marker for early screening of sarcopenia in kidney transplant recipients. However, its predictive value needs to be verified further by large-sample cohort studies, and thus to better promote early clinical screening of sarcopenia in kidney transplant recipients to effectively improve their quality of life and prognosis

Ultrastructural Aspects of Rat Renal Tubular Epithelium in Vitro: Scanning Electron Microscopy （SEM） Analyses at Various Stages of Culture

Renal tubular epithelial cells are capable of regenerating themselves after severe injury. In cellular regeneration, cell migration plays a crucial role. Previous studies have demonstrated that changes in cellular morphology, including the formation of lamellipodia, are very important in cell migration. However, the ultrastructure of a migrating cell largely remains undescribed. In the present study, we applied scanning electron microscopy techniques to cultured cells, and observed fine ultrastructural changes in cultured renal tubular epithelium during migration. We found that there were small sphere-like structures and/or microvilli on the cell surface of floating epithelia, that leaf-like structures closely resembling lamellipodia developed circumferentially in the early stages of culture, and that long and branched filopodia developed that were much larger in diameter compared to filopodia at earlier stages. To the best of our knowledge, these findings are new, and may contribute to the development of a new therapeutic strategy for severe tubular damage of the kidney

Low Protein Diet Inhibits Uric Acid Synthesis and Attenuates Renal Damage in Streptozotocin-Induced Diabetic Rats

Aim. Several studies indicated that hyperuricemia may link to the worsening of diabetic nephropathy (DN). Meanwhile, low protein diet (LPD) retards exacerbation of renal damage in chronic kidney disease. We then assessed whether LPD influences uric acid metabolism and benefits the progression of DN in streptozotocin- (STZ-) induced diabetic rats. Methods. STZ-induced and control rats were both fed with LPD (5%) and normal protein diet (18%), respectively, for 12 weeks. Vital signs, blood and urinary samples for UA metabolism were taken and analyzed every 3 weeks. Kidneys were removed at the end of the experiment. Results. Diabetic rats developed into constantly high levels of serum UA (SUA), creatinine (SCr) and 24 h amounts of urinary albumin excretion (UAE), creatintine (UCr), urea nitrogen (UUN), and uric acid (UUA). LPD significantly decreased SUA, UAE, and blood glucose, yet left SCr, UCr, and UUN unchanged. A stepwise regression showed that high UUA is an independent risk factor for DN. LPD remarkably ameliorated degrees of enlarged glomeruli, proliferated mesangial cells, and hyaline-degenerated tubular epithelial cells in diabetic rats. Expression of TNF-α in tubulointerstitium significantly decreased in LPD-fed diabetic rats. Conclusion. LPD inhibits endogenous uric acid synthesis and might accordingly attenuate renal damage in STZ-induced diabetic rats

Identification of Isoflavonoid Biosynthesis-Related R2R3-MYB Transcription Factors in Callerya speciosa (Champ. ex Benth.) Schot Using Transcriptome-Based Gene Coexpression Analysis

The R2R3-MYB family is one of the largest plant transcription factor (TF) families playing vital roles in defense, plant growth, and secondary metabolism biosynthesis. Although this gene family has been studied in many species, isoflavonoid biosynthesis-related R2R3-MYB TFs in Callerya speciosa (Champ. ex Benth.) Schot, a traditional Chinese medicinal herb, are poorly understood. Here, a total of 101 R2R3-MYB TFs were identified from C. speciosa transcriptome dataset. 25 clades divided into five functional groups were clustered based on the sequence similarity and phylogenetic tree. Conserved motifs and domain distribution, expression patterns, and coexpression networks were also employed to identify the potential R2R3-MYB TFs in the regulation of isoflavonoid biosynthesis. In silico evaluation showed that the deduced R2R3-CsMYB proteins contain highly conserved R2R3 repeat domain at the N-terminal region, that is the signature motif of R2R3-type MYB TFs. Eight potential TFs (CsMYB17, CsMYB36, CsMYB41, CsMYB44, CsMYB45, CsMYB46, CsMYB72, and CsMYB81) had high degrees of coexpression with four key isoflavonoid biosynthetic genes (CsIFS, CsCHS7, CsHID-1, and CsCHI3), in which CsMYB36 as a potential regulator possessed the highest degree. HPLC analysis showed that formononetin and maackiain contents were significantly increased during the development of tuberous roots, which might be controlled by both related R2R3-CsMYBs and structural genes involved in the isoflavonoid biosynthesis pathway. The transcriptome data were further validated by reverse transcription real-time PCR (RT-qPCR) analysis, and similar expression profiles between TFs and key structural genes were identified. This study was the first step toward the understanding of the R2R3-MYB TFs regulating isoflavonoid biosynthesis in C. speciosa. The results will provide information for further functional analysis and quality improvement through genetic manipulation of these potential R2R3-CsMYB genes in C. speciosa

Integrated transcriptomics and metabolomics analysis provides insights into aromatic volatiles formation in Cinnamomum cassia bark at different harvesting times

Abstract Background Cinnamomum cassia Presl, classified in the Lauraceae family, is widely used as a spice, but also in medicine, cosmetics, and food. Aroma is an important factor affecting the medicinal and flavoring properties of C. cassia, and is mainly determined by volatile organic compounds (VOCs); however, little is known about the composition of aromatic VOCs in C. cassia and their potential molecular regulatory mechanisms. Here, integrated transcriptomic and volatile metabolomic analyses were employed to provide insights into the formation regularity of aromatic VOCs in C. cassia bark at five different harvesting times. Results The bark thickness and volatile oil content were significantly increased along with the development of the bark. A total of 724 differentially accumulated volatiles (DAVs) were identified in the bark samples, most of which were terpenoids. Venn analysis of the top 100 VOCs in each period showed that twenty-eight aromatic VOCs were significantly accumulated in different harvesting times. The most abundant VOC, cinnamaldehyde, peaked at 120 months after planting (MAP) and dominated the aroma qualities. Five terpenoids, α-copaene, β-bourbonene, α-cubebene, α-funebrene, and δ-cadinene, that peaked at 240 MAP could also be important in creating C. cassia’s characteristic aroma. A list of 43,412 differentially expressed genes (DEGs) involved in the biosynthetic pathways of aromatic VOCs were identified, including phenylpropanoids, mevalonic acid (MVA) and methylerythritol phosphate (MEP). A gene-metabolite regulatory network for terpenoid and phenylpropanoid metabolism was constructed to show the key candidate structural genes and transcription factors involved in the biosynthesis of terpenoids and phenylpropanoids. Conclusions The results of our research revealed the composition and changes of aromatic VOCs in C. cassia bark at different harvesting stages, differentiated the characteristic aroma components of cinnamon, and illuminated the molecular mechanism of aroma formation. These foundational results will provide technical guidance for the quality breeding of C. cassia

Relationship between total ghrelin and nutritional parameters in maintenance hemodialysis patients

Ghrelin is regarded to be correlated to nutrition status. To verify this relationship, 30 patients on hemodialysis(HD), 18 patients with chronic kidney disease(CKD) and 18 healthy volunteers(Control) were involved in this observational study. Total plasma ghrelin(ELISA) and nutritional parameters (including biochemical index, body composition, and nutrition risk screening score 2002, NRS2002) were measured. Data were showed by Mean±SD, probability values <0.05 were considered significant. Statistical analysis was determined using SPSS 15.0. Ghrelin levels was significantly increased in HD patients (4.55±2.34ng/ml (pre-HD), p<0.0001) than in CKD(2.32±1.32ng/ml) and Control (1.99±0.83ng/ml),and declined after HD(2.27±1.12ng/ml, p<0.0001). In HD group, plasma ghrelin levels were negatively correlated with pre-albumin(PA, r=-0.461,P=0.010). When all participants combined together, the plasma ghrelin levels was positively correlated with serum creatinine(r=0.426,P=0.0001) and urea nitrogen(r=0.366,P=0.003),but negatively correlated with e-GFR(r=-0.411,P=0.001), PA(r=-0.321s,P=0.009) and lymphocyte(r=-0.417,P=0.0001). No relationship was showed between ghrelin and BMI, NRS2002 in HD group. In conclusion, total ghrelin levels was elevated in HD patients, and negatively correlated with pre-albumin, and negatively correlate with PA,lymphocyte in all participants. A future study with the stratification of HD patients according to their appetite and body composition may help to further evaluation

Febuxostat Attenuates Renal Damage besides Exerting Hypouricemic Effect in Streptozotocin-Induced Diabetic Rats

Aim. In this study, we aimed to investigate the effects of febuxostat, a novel inhibitor of xanthine oxidase (XO), on renal damage in streptozotocin- (STZ-) induced diabetic rats. Methods. Diabetes was induced by the intraperitoneal injection of STZ in male Sprague-Dawley rats. Sham-injected rats served as controls. The control and diabetic rats were treated with and without febuxostat for 8 weeks, respectively. Fasting blood and 24-h urine samples were collected every 4 weeks. Rat livers were extracted for detecting gene expression, content, and bioactivity of XO. Results. Diabetic rats showed significantly increased serum uric acid (SUA), serum creatinine (SCr), and urea nitrogen (BUN) levels. Daily urinary albumin (UAE), uric acid (UUA), and creatinine (UCr) excretion were also significantly increased in these rats. In diabetic rats, at week 8, febuxostat decreased SUA by 18.9%, while UAA was increased by 52.0%. However, UCr and urinary urea nitrogen (UUN) levels remained unchanged, while SCr and BUN levels decreased by >30% in these rats. Although hepatic gene expression, content, and activity of XO increased significantly in diabetic rats, febuxostat only slightly decreased its content. Conclusions. Febuxostat significantly attenuated renal damage in STZ-induced diabetic rats in addition to exerting hypouricemic effect

Genome-wide characterization of the bHLH gene family in Gynostemma pentaphyllum reveals its potential role in the regulation of gypenoside biosynthesis

Abstract Background Gynostemma pentaphyllum, an ancient Chinese herbal medicine, serves as a natural source of gypenosides with significant medicinal properties. Basic helix-loop-helix (bHLH) transcription factors play pivotal roles in numerous biological processes, especially in the regulation of secondary metabolism in plants. However, the characteristics and functions of the bHLH genes in G. pentaphyllum remain unexplored, and their regulatory role in gypenoside biosynthesis remains poorly elucidated. Results This study identified a total of 111 bHLH members in G. pentaphyllum (GpbHLHs), categorizing them into 26 subgroups based on shared conserved motif compositions and gene structures. Collinearity analysis illustrated that segmental duplications predominately lead to the evolution of GpbHLHs, with most duplicated GpbHLH gene pairs undergoing purifying selection. Among the nine gypenoside-related GpbHLH genes, two GpbHLHs (GpbHLH15 and GpbHLH58) were selected for further investigation based on co-expression analysis and functional prediction. The expression of these two selected GpbHLHs was dramatically induced by methyl jasmonate, and their nuclear localization was confirmed. Furthermore, yeast one-hybrid and dual-luciferase assays demonstrated that GpbHLH15 and GpbHLH58 could bind to the promoters of the gypenoside biosynthesis pathway genes, such as GpFPS1, GpSS1, and GpOSC1, and activate their promoter activity to varying degrees. Conclusions In conclusion, our findings provide a detailed analysis of the bHLH family and valuable insights into the potential use of GpbHLHs to enhance the accumulation of gypenosides in G. pentaphyllum

Effect of Maternal Intestinal Floras on Neonatal Birth Weight

Background Birth weight is closely related to individual health. Low birth weight is a high-risk factor for early neonatal death. Macrosomia is associated with higher risk of maternal and infant complications and various chronic diseases in adulthood. Therefore, it is very important to identify the influencing factors of neonatal birth weight. Objective To investigate the relationship between maternal intestinal flora and neonatal birth weight. Methods Participants were 516 singleton mothers and their babies〔24 with low birth weight (LW group) , 479 with normal birth weight (NW group) and 13 with macrosomia (OW group) 〕 delivered at term in Guangzhou Women and Children&apos;s Medical Center from January to September 2017. Maternal intestinal flora and clinical laboratory test parameters were collected, and the composition and diversity of intestinal flora were analyzed using QIIME. LEfSe analysis was used to compare the relative abundance of intestinal flora at the genus level of the mothers of three groups of babies to identify the flora with significant intergroup differences. MaAslin was used to assess associations of maternal laboratory test parameters and microbial genera. The Boruta was used to build models for predicting three types of neonatal birth weight using maternal laboratory test parameters and intestinal flora OTUs, to assess the association of maternal intestinal floras and neonatal birth weight. Results The analysis of maternal intestinal floras showed that the abundance of Firmicutes was the highest at the phylum level, and Faecalibacterium was significantly enriched at the genus level. There were no significant differences in Simpson&apos;s Diversity Index and Shannon Diversity Index at the phylum level across the maternal intestinal floras of three groups of babies (P&gt;0.05) . LEfSe analysis found that compared with intestinal flora of mothers of LW group, the intestinal flora of mothers of NW group showed significantly enriched Streptococcus and Roseburia (P&lt;0.05) , and significantly reduced abundance of Bacillaceae, Raphanus, Methanosphaera, Barnesiella and Paraprevotella (P&lt;0.05) , while the intestinal flora of mothers of OW group demonstrated significantly enriched Closrtidiaceae and Alistipes as well as significantly reduced abundance of Barnesiella (P&lt;0.05) . Compared with intestinal flora of mothers of NW group, the intestinal flora of mothers of OW group indicated significantly enriched Megamonas, Coprococcus, Veillonellaceae, cc-115, Closrtidiaceae and Alistipes, and significantly reduced abundance of Blautia and Eggerthella (P&lt;0.05) . The area under ROC curve (AUC value) based on laboratory test parameter OTUs model and intestinal flora OTUs model was 0.62 and 0.77, respectively, in discriminating LW from NW, and was 0.65, and 0.78 respectively, in discriminating OW from NW. Conclusion Neonatal birth weight varied by the features of maternal intestinal floras. The OTUs model based on maternal intestinal flora could distinguish the neonatal birth weight. Maternal intestinal flora may be a good predictor of neonatal birth weight