Study on the mechanism of hirudin multi target delaying renal function decline in chronic kidney disease based on the "gut-kidney axis" theory.

The disorder of the "gut-kidney axis" exacerbates renal function decline in chronic kidney disease (CKD), and current CKD therapy is insufficient to address this issue. Hirudin has a palliative effect on the decline of renal function. However, whether hirudin can delay CKD by regulating the "intestinal renal axis" disorder remains unclear. Unilateral ureteral ligation (UUO) induced CKD rat model, and the rats were treated with bifidobacterium and hirudin for 36 days. After 14 and 36 days of modeling, kidney and colon tissues were collected for pathology, western blot (WB) assay, and quantitative real-time PCR (qPCR) detection. Serum samples were collected for renal function testing. Fecal samples were used for 16S rRNA sequencing and research on fecal bacterial transplantation. Lipopolysaccharide combine with adenosine 5'-triphosphate (LPS + ATP)-induced intestinal epithelial cell injury was treated with a nod-like receptor pyrin domain-associated protein 3 (NLRP3) inhibitor and hirudin. Protein expression was detected using WB and qPCR. The kidneys and colons of the CKD rats exhibited varying degrees of lesions. Creatinine (CRE), blood urea nitrogen (BUN), N-acetyl-ß-D-glucosidase (NAG) enzyme, and serum uremic toxins were elevated. The expression of claudin-1 and occludin was decreased, NLRP3 inflammatory-related proteins were increased, and the gut microbiota was disrupted. These pathological changes were more pronounced after 36 days of modeling. Meanwhile, high-dose hirudin treatment significantly improved these lesions and restored the intestinal flora to homeostasis in CKD rats. In vitro, hirudin demonstrated comparable effects to NLRP3 inhibitors by upregulating claudin-1 and occludin expression, and downregulating NLRP3 inflammatory-related proteins expression. The dysbiosis of the gut microbiota and impaired intestinal epithelial barrier function in CKD are associated with renal dysfunction in CKD. Hirudin delays the progression of CKD by regulating the disorder of the "gut-kidney axis" and inhibiting the activation of the NLRP3-ASC-caspase-1 pathway.

Clinical characteristics of hypercalcemic crises in a tertiary children's hospital.

BACKGROUND: Hypercalcemic crisis is considered a critical and fatal endocrine condition. To date, few reports have focused on hypercalcemic crises in children. AIM: To explore the etiology and identify the clinical characteristics related to hypercalcemic crises in children. METHODS: We enrolled 101 children diagnosed with hypercalcemia between January 1, 2016 and December 31, 2021, admitted to the Children's Hospital of Chongqing Medical University. Electronic medical records were reviewed to determine the causes and clinical characteristics of hypercalcemic crises. RESULTS: Hypercalcemic crises occurred in 28 admissions during the 6-year period; 64% of the patients enrolled in the study were infants. The mean corrected total serum calcium was 4.6 ± 0.2 mmol/l. Tumor and hereditary diseases were found in 12 (43%) and 7 (25%) patients, respectively. The ratio of iatrogenic factors was 11% (3/28), and all 3 patients received a blood transfusion. The incidence of poor prognosis in the tumor cases was 50%. Timely intervention including hemodialysis, pamidronate, and etiological treatment was effective in decreasing calcium levels. CONCLUSION: Hypercalcemic crisis is a serious electrolyte disturbance that has the potential for high mortality. The main causes are tumors and hereditary diseases in children. The lack of unique characteristics makes it difficult to recognize by medical caregivers. Early diagnosis and timely intervention could improve the prognosis.

Hirudin attenuates puromycin aminonucleoside-induced glomerular podocyte injury by inhibiting MAPK-mediated endoplasmic reticulum stress.

Damage to podocytes is an important determinant of renal pathology. The puromycin aminonucleoside (PAN) mice nephropathy model is commonly used in the study of renal disease with podocyte injury. Hirudin has a broad nephroprotective effect and has been shown to treat renal interstitial fibrosis in previous studies. Mice were given PAN by gavage to prepare animal models, and MPC5 cells were incubated with PAN in vitro. Twenty-four hours urine was collected for analysis of urinary protein levels. Renal pathological changes were observed by hematoxylin and eosin staining. Immunofluorescence detection of nephrin in kidney tissues and cells. Apoptosis was analyzed with over TUNEL. Cytoskeleton, endoplasmic reticulum stress (ERS), p38 MAPK signaling, and apoptosis-related proteins were assessed by western blot analysis. The data suggested that hirudin attenuated reduced renal injury and increased urine protein in PAN mice. Hirudin also attenuated cytoskeletal protein (synaptopodin, nephrin, and podocin) disruption, ERS activation, and apoptosis in PAN mice and PAN-induced podocytes. In addition, hirudin inhibited the expression of p38 MAPK signaling key proteins upregulated by PAN, thereby suppressing ERS. The p38 MAPK agonist was able to partially antagonize the inhibition of p38 MAPK signaling by hirudin in PAN-induced podocytes, thereby reactivating the ERS inhibited by hirudin, promoting cytoskeletal protein degradation and increasing the level of apoptosis. In conclusion, hirudin could decrease podocyte injury by inhibiting p38 MAPK signaling-mediated ERS, resulting in the protection of the kidney from PAN damage. These findings may provide an experimental basis for hirudin treatment of podocyte injury diseases.

Hirudin, a thrombin inhibitor, attenuates TGF-ß-induced fibrosis in renal proximal tubular epithelial cells by inhibition of protease-activated receptor 1 expression via S1P/S1PR2/S1PR3 signaling.

Renal interstitial fibrosis (RIF) is the final common outcome of numerous chronic kidney diseases, contributing to end-stage renal disease. Hirudin, a thrombin inhibitor, has attracted increased attention as a potential treatment approach for renal fibrosis. The present study aimed to investigate the molecular mechanism underlying the effect of hirudin on fibrosis in renal proximal tubular epithelial cells. An in vivo mouse RIF model established using unilateral ureteral obstruction (UUO) and an in vitro of RIF using the renal tubular epithelial cell line HK-2 treated with TGF-ß were used. Expressions of sphingosine-1-phosphate (S1P) receptors (S1PR)1-4 and protease-activated receptor 1 (PAR1) were measured by reverse transcription-quantitative PCR and western blotting in mice with UUO and TGF-ß induced HK-2 cells. Western blotting was used to detect the expression of N-cadherin, Slug, E-cadherin, Collagen IV, fibronectin, MMP9 and monocyte chemoattractant protein-1. Immunofluorescence staining was conducted to measure α-SMA level expression. The results demonstrated that the expression levels of S1PR1, S1PR2, S1PR3, S1PR4 and PAR1 were upregulated in both TGF-ß-induced HK-2 cells and renal tissues from mice with unilateral ureteral ligation. Notably, hirudin inhibited TGF-ß-induced PAR1, S1PR2 and S1PR3 upregulation in both HK-2 cells and renal tissues. Additionally, the inhibition of S1PR2 and S1PR3 resulted in PAR1 downregulation. Furthermore, treatment with S1P and PAR1 agonists abolished the effect of hirudin on the expression of EMT, fibrosis-related proteins and monocyte chemoattractant protein 1. In conclusion, hirudin attenuated TGF-ß-induced fibrosis in proximal renal tubular epithelial HK-2 cells by inhibiting PAR1 expression via the S1P/S1PR2/S1PR3 signaling pathway. Therefore, hirudin may be considered as a promising therapeutic agent for RIF.

Higenamine inhibits apoptosis and maintains survival of gastric smooth muscle cells in diabetic gastroparesis rat model via activating the ß2-AR/PI3K/AKT pathway.

Diabetic gastroparesis (DGP) is a common complication of diabetes mellitus (DM). The numerous clinical symptoms of DGP and the great cost on the treatment of DGP seriously lowered the patients' life quality. However, the pathogenic mechanism of DGP is still elusive till now. In this study, we aimed to explore the effect of higenamine on the proliferation and apoptosis of gastric smooth muscle cells (SMCs) in DGP rat model. The DGP rat model was built by intraperitoneal injection of Streptozotocin (STZ) into male Sprague-Dawley (SD) rats. Compared with the healthy control group, the level of DGP indicator c-kit was strongly suppressed and the level of Gsα was largely elevated in the STZ-induced model group. By contrast, the addition of higenamine obviously counteracted the effect of STZ on the expression of c-kit and Gsα. Besides that, higenamine improved the decreased emptying ability of the stomach. In addition, the number of gastric SMCs was strongly decreased and cell morphology became irregular in STZ-induced model group. The treatment of higenamine weakened the harm of STZ on the number and morphology of gastric SMCs. Beyond that, higenamine promoted gastric SMCs proliferation and inhibited gastric SMCs apoptosis in DGP model. Further research revealed that higenamine regulated cell proliferation and apoptosis via activating the ß2-AR/PI3K/AKT pathway. Taken together, our research revealed that higenamine maintained the survival of gastric SMCs in DGP rat model via the ß2-AR/PI3K/AKT pathway, providing a new sight for the treatment of DGP.

Waist circumference and waist-to-height ratio in Han Chinese children living in Chongqing, south-west China.

OBJECTIVE: To derive age- and sex-specific reference values for waist circumference (WC) and waist-to-height ratio (WHtR) for Han Chinese children and adolescents and to establish the prevalence of excess central adiposity in our study population. DESIGN: Cross-sectional study of schoolchildren attending randomly selected primary and secondary schools in south-west China in October 2003 and April 2004. Anthropometry was measured using standard procedures. The LMS method was used to construct smoothed WC and WHtR percentile curves. Overweight and obesity were defined by the International Obesity Task Force (IOTF) criteria and the Working Group on Obesity in Children. Excess central adiposity fat was defined by previously published WC cut-points and a WHtR ≥ 0.5. SETTING: Primary and secondary schools in Chongqing, south-west China. SUBJECTS: A total of 7326 (49.2 % boys) Han Chinese students at 5-17 years old. RESULTS: On the basis of the IOTF criteria, 26.4 % of boys were overweight or obese compared with 16.4 % of girls (P < 0.001). WC cut-points identified 31 % of boys and 28 % of girls as having excess central adiposity, whereas using the WHtR criterion, 14.8 % of boys and 5.6 % of girls were identified. Young boys (5-12 years) had a significantly (P < 0.001) higher WHtR than girls. CONCLUSIONS: We have constructed WC and WHtR percentile curves for Han Chinese children and adolescents living in Chongqing. Our measurements were based on a student population with a relatively high rate of overweight and obesity. These data will provide a point of reference for future studies measuring the prevalence of overweight and obesity in China.