Taxifolin (TAX) is a natural compound known for its liver protection effect, but the mechanism remains unknown. Phosphorylated proteomics analyses discovered that the phosphorylation level of NDRG1 at T328 was a key event of TAX-improved liver fibrosis. We established models with NDRG1 knockout (KO) in vivo and in vitro, demonstrating that NDRG1 KO attenuated the development of hepatocyte injury, and combining NDRG1 KO and TAX administration did not result in a reduction in protection against liver injury. Cellular thermal shift assay and surface plasma resonance analysis showed that TAX directly binds to NDRG1 rather than its upstream kinase, subsequently demonstrating that TAX regulated phosphorylation of NDRG1 at T328 through binding to its C289 site. NDRG1 T328A (phosphorylated mutation) and T328E (mimic phosphorylation) in vivo and in vitro confirmed that pNDRG1_T328 exacerbates hepatocyte injury along with DNA damage, inflammatory response, and apoptosis, thereby contributing to hepatic stellate cells (HSCs) activation. In contrast, TAX can inhibit the above pathological abnormalities and block hepatocyte injury-triggered HSCs activation and fibrosis. Overall, TAX is a potent liver protection drug primarily targeting NDRG1 and inhibiting pNDRG1_T328 in hepatocytes.

Objective:To investigate the effects of human glycolipid transfer protein(GLTP)on proliferation,migration and invasion of pancreatic cancer(PC)cells,and to elucidate their mechanisms.Methods:The difference in the expression levels of GLTP proteins between PC tissue and normal pancreas tissue was analyzed by University of Alabama at Birmingham Cancer Data Analysis Platform(UALCAN)Database,as well as the difference in the expression levels of GLTP protein between PC tissue and normal pancreas tissue of the PC patients with different clinicopathological characteristics.The PANC-1 cells were cultured in vitro and divided into control group(transfected with pFLAG-CMV4 plasmid)and GLTP-overexpression(GLTP-OE)group(transfected with pFLAG-GLTP plasmid).The stably GLTP transfected cells were established using the antibiotic screening method.Knock-down experiments were performed using non-specific siRNA transfected PANC-1 cells as control group and si-GLTP transfected PANC-1 cells as si-GLTP group.Western blotting method was used to detect the expression of GLTP protein in the cells in various groups,cell counting kit-8(CCK-8)method was used to detect the proliferation activities of PANC-1 cells,clone formation assay was used to detect the number of clone formation,and Transwell chamber assay were used to detect the numbers of migration and invasion cells in various groups.Transcriptomics sequencing analyses were conducted to assess the possible mechanism of GLTP in the PANC-1 cells.Western blotting method was employed to detect the expression levels of phosphatidylinositol 3-kinase(PI3K),phosphorylated PI3K(p-PI3K),protein kinase B(Akt),and phosphorylated Akt(p-Akt)proteins in the PANC-1 cells in various groups;Real-time fluorescence quantitative PCR(RT-qPCR)was used to assess the expression levels of amphiregulin(AREG)and kinase insertion domain receptor(KDR)mRNA in the cells in various groups.The mice were randomly divided into control group(injected with pFLAG-GMV4 transfected PANC-1 cells)and experimental group(injected with pFLAG-GLTP stably transfected PANC-1 cells),and the subcutaneously transplanted tumor models were prepared;the volumes and weights of the transplanted tumors of the mice in two groups were measured.Results:UALCAN database analysis showed that the expression level of GLTP protein in PC tissue was lower than that in normal pancreas tissue(P＜0.01),and there were statistically significant differences in the GLTP protein expression between PC tissue and normal pancreas tissue of the PC patients with different cancer stages(P＜0.05),tumor grades(P＜0.05),ages(P＜0.001),and genders(P＜0.05).Compared with control group,the proliferation activity(P＜0.01)and the number of clone formation(P＜0.001)of the cells in GLTP-OE group were decreased,while the numbers of migration cells(P＜0.001)and invasion cells(P＜0.01)were decreased.In the knock-down experiment,compared with control group,the proliferation activity(P＜0.01)and the number of clone formation(P＜0.05)of the cells in GLTP-OE group were increased,while the numbers of migration cells(P＜0.001)and invasion cells(P＜0.001)were increased.Compared with control group,the tumor weight and volume of the mice in experimental group were decreased(P＜0.01),following the injection of tumor cells for a period of four weeks.In the over-expression experiment,compared with control group,the expression levels of p-PI3K(P＜0.01),p-Akt-S473(P＜0.01),and p-Akt-T308(P＜0.05)proteins in the cells in GLTP-OE group were decreased;the expression levels of AREG(P＜0.01)and KDR(P＜0.01)mRNA were decreased.In the knock-down experiment,compared with control group,the expression levels of p-PI3K(P＜0.01),p-Akt-S473(P＜0.01),and p-Akt-T308(P＜0.01)in the cells in si-GLPT group were increased,and the expression levels of AREG(P＜0.01)and KDR(P＜0.05)mRNA were increased.Conclusion:Low expression levels of GLTP in PC tissue are present.The over-expression of GLTP can inhibit the proliferation,migration and invasion of PANC-1 cells,as well as the growth of subcutaneously transplanted tumors in the nude mice;its possible mechanism may be related to decreasing the activity of the PI3K/Akt signaling pathway.

Epidural labor analgesia aims to provide effective medical services to alleviate labor pain in parturients, while adhering to the principles of voluntary participation and clinical safety. In 2018, Peking Union Medical College Hospital(PUMCH)became one of the first pilot units for labor analgesia in China, and has achieved satisfactory results in high-quality development of labor analgesia. This article mainly introduces the achievements and experience of labor analgesia at PUMCH, including: (1) prioritizing maternal and infant safety, arranging personnel rationally, and developing standardized treatment processes through multidisciplinary collaboration to ensure safe and comfortable childbirth; (2) leveraging the hospital's comprehensive capabilities in emergency treatment, and improving collaborative rescue plans for critically ill parturients and newborns; (3) implementing advanced teaching methods to effectively train and conduct simulated drills for labor analgesia and rescue of critically ill parturients; (4) conducting patient education and informative lectures to help parturients acquire a scientific understanding of labor analgesia. We hope that this experience can provide reference and inspiration for other hospitals.

Renal fibrosis is a devastating consequence of progressive chronic kidney disease,representing a major public health challenge worldwide.The underlying mechanisms in the pathogenesis of renal fibrosis remain unclear,and effective treatments are still lacking.Renal tubular epithelial cells(RTECs)maintain kidney function,and their dysfunction has emerged as a critical contributor to renal fibrosis.Cellular quality control comprises several components,including telomere homeostasis,ubiquitin-proteasome system(UPS),autophagy,mitochondrial homeostasis(mitophagy and mitochondrial metabolism),endoplasmic reticulum(ER,unfolded protein response),and lysosomes.Failures in the cellular quality control of RTECs,including DNA,protein,and organelle damage,exert profibrotic functions by leading to senescence,defective autophagy,ER stress,mitochondrial and lysosomal dysfunction,apoptosis,fibro-blast activation,and immune cell recruitment.In this review,we summarize recent advances in un-derstanding the role of quality control components and intercellular crosstalk networks in RTECs,within the context of renal fibrosis.

Objective:To summarize the precision fluid management of patients with severe blast injury in the emergency intensive care unit, so as to help patients smoothly pass through the dangerous period and recover smoothly.Methods:Based on the experience of fluid management in 6 patients admitted to the Second Affiliated Hospital Zhejiang University School of Medicine in the tanker truck explosion on 14 June, 2020. The main measures included: fluid volume management and dynamic adjustment; assessment of intake, output and urine volume, and dynamic adjustment of infusion volume and speed; monitoring of pulmonary oxygenation and timely adjustment of fluid resuscitation strategies; monitoring indexes and providing nursing care strategies for fluid management.Results:Finally, among 6 patients with severe blast injury, 5 patients were discharged from the hospital with follow-up treatment after they suffered from the shock and infection phases and refined fluid management, 1 patient died due to severe injury and ineffective rescue.Conclusions:Adopting individualized, phased, and refined liquid management strategy has clinical significance for patients with severe blast injury to smoothly pass the risk period.

Clouston syndrome (OMIM #129500), also known as hidrotic ectodermal dysplasia type 2, is a rare autosomal dominant skin disorder. To date, four mutations in the GJB6 gene, G11R, V37E, A88V, and D50N, have been confirmed to cause this condition. In previous studies, the focus has been mainly on gene sequencing, and there has been a lack of research on clinical manifestations and pathogenesis. To confirm the diagnosis of this pedigree at the molecular level and summarize and analyse the clinical phenotype of patients and to provide a basis for further study of the pathogenesis of the disease, we performed whole-exome and Sanger sequencing on a large Chinese Clouston syndrome pedigree. Detailed clinical examination included histopathology, hair microscopy, and scanning electron microscopy. We found a novel heterozygous missense variant (c.134G>C:p.G45A) for Clouston syndrome. We identified a new clinical phenotype involving all nail needling pain in all patients and found a special honeycomb hole structure in the patients' hair under scanning electron microscopy. Our data reveal that a novel variant (c.134G>C:p.G45A) plays a likely pathogenic role in this pedigree and highlight that genetic testing is necessary for the diagnosis of Clouston syndrome.
Humans ; Connexin 30/genetics* ; Connexins/genetics* ; East Asian People ; Ectodermal Dysplasia/pathology* ; Phenotype

Objective To study the changes of renal cortex angiotensin Ⅱ (AngⅡ) expression and podocyte autophagy in aging rats with normotension versus hypertension,to further investigate the possible mechanism of renal injury in hypertension during aging.Methods Normotensive Wistar-Kyoto-rats (WKYs) were allocated to three groups by age of 3-month-old group,13-month-old group,22-month-old group.Gender,age and number-matched spontaneously hypertensive rats (SHRs) served as controls.Blood pressure was monitored.Levels of urinary albumin,urinary creatinine,serum creatinine (SCR),blood urea nitrogen (BUN),Ang Ⅱ in serum and in renal cortex were detected.The kidney ultrastructural changes and podocyte autophagosomes were observed under light and transmission electron microscopy.Expressions of nephrin,LC3B Ⅱ,Atg5 and p62 in glomeruli were analyzed by Western blotting.Results Blood pressure was significantly higher in SHRs than in WKYs (P＜0.05).During aging from 3-month-old to 13-month-old and to 22 monthold group,the urinary albumin/creatinine ratio was increased significantly in SHRs with hypertension and in WKYs with normotension [WKY:(0.12±0.01) g/mmol,(0.14±0.04) g/mmol vs.(0.34± 0.05) g/mmol;in SHR:(0.29±0.04) g/mmol,(0.31±0.05) g/mmol vs.(0.72±0.16) g/mmol,P＜0.05],but SCR and BUN levels had no significant difference in SHRs and WKYs during aging (both P＞0.05).And Ang Ⅱ levels in both serum and renal cortex had no significant change in normotension group during aging three time points (both P＞0.05).An age-dependent decrease of Ang Ⅱ level in renal cortex appeared during aging three time points in hypertensive rats [SHR:(0.02 ±0.00) μg/L,(0.02±0.00) μg/L vs.(0.01±0.00) μg/L,P＜0.05],but serum level of Ang Ⅱ had no significant difference during aging three time points (P＞ 0.05) in hypertension group.The structural changes,including glomerulosclerosis,tubular atrophy and interstitial fibrosis were observed during aging three time points both in normotensive and hypertensive rats,but these pathological changes were more serious in hypertensive rats.Autophagosomes relatively accumulated in aging normotensive rats.The protein expressions of LC3B Ⅱ,Atg5 and p62 were increased in normotensive rats during aging (all P＜0.05).While the protein expressions of nephrin,LC3B Ⅱ,Atg5 and p62 were decreased in aging hypertensive rats (all P ＜ 0.05).Conclusions Ang Ⅱ expression level in renal cortex is decreased during aging in hypertensive rats.Podocyte autophagic activity is relatively decreased during aging in normotensive rats,but is relatively increased during aging in hypertensive rats.Ang Ⅱ-induced podocyte autophagy may be involved in the pathogenesis of renal injury in hypertension during aging.

Objective To examine the effects of benazepril and losartan on glomerular podocyte autophagy in aged spontaneously hypertensive rats (SHRs) and investigate the underlying mechanisms of renal-protective effects of angiotensin-converting enzyme inhibitors (ACEI)/angiotensin receptor blockers (ARB).Methods Wistar-Kyoto rats (WKYs) were used as the normal control (NORM) group (2 ml physiological saline per day).SHRs were randomly divided into 4 groups:the CTRL group (2 ml physiological saline per day),the ACEI group (10 mg · kg-1 · d-1),the ARB group (30 mg· kg-1 ·d-1) and the combined group (10 mg· kg-1 · d-1 benazepril and 30 mg· kg-1 · d-1),with six 18-month-old make rats in each group.The experiments were conducted during a 4-month period.Blood pressure was monitored regularly.At the end of the experiments,we measured the levels of urine protein,urine creatinine,serum creatinine (SCR),blood urea nitrogen (BUN),and serum and renal cortex angiotensin Ⅱ (AngⅡ).Ultrastructural changes in the kidney were examined under light and transmission electron microscopy.The expressions of nephrin,LC3BⅡ,Atg5 and p62 in the glomerulus were analyzed by Western blot analysis.Results After treatment,the blood pressure and the urine albumin/creatinine ratio of the four SHR groups were still significantly higher than those of the NORM group,but the blood pressure and the urine albumin/creatinine ratio of the ARB group and the combined group were significantly lower than those of the CTRL group (all P＜ 0.05);There were no significant differences in SCR and BUN levels among these five groups (P＞ 0.05);The level of serum AngⅡ of the combined group was significantly higher than that of the CTRL group [CTRL (0.08±0.00) μg/L,Combined (0.12±0.01) μg/L,P＜0.05];The levels of cortex AngⅡ of the four SHR groups were significantly lower than those of the NORM group,while the level of cortex AngⅡ of the ARB group was significantly higher than that of the CTRL group (all P＜0.05);Renal ultrastructural examination revealed shrunken glomeruli,fused or effaced epithelial cell foot processes,and focal atrophy of renal tubules in the four SHR groups.These pathological changes were more serious in the CTRL group but less so in the combined group.There were significantly more autophagosomes in the NORM group and the combined group than in the CTRL group (P＜0.05).Compared with the NORM group,the expressions of nephrin,LC3BⅡ,Atg5 and p62 in the CTRL group were suppressed significantly (P ＜ 0.05).The expressions of nephrin,LC3BⅡ and Atg5 in the ACEI group and the expressions of nephrin,LC3BⅡ,Atg5 and p62 in the ARB group and the combined group were higher than in the CTRL group (P＜0.05).Conclusions ACEI/ARB can decrease the autophagic activity of glomerular podocytes.The renal-protective effects of ACEI/ARB may be mediated by glomerular podocyte autophagy,which is induced by AngⅡ.