OBJECTIVE To establish an HPLC method for the separation of enantiomers of farrerol, and apply it to the determination of the content of enantiomers in Rhododendri Daurici Folium and Rhododendron Micranthum. METHODS HPLC was used to separate the farrerol enantiomers, and the chromatographic conditions of chiral column type, mobile phase ratio, flow rate, and column temperature were optimized. The thermodynamic separation of farrerol enantiomers was discussed. Thermodynamic parameters such as enthalpy change, entropy change, enthalpy change and entropy change were calculated. And the contents of two enantiomers in Rhododendri Daurici Folium and Rhododendron Micranthum were determined under the optimum resolution conditions. RESULTS The optimum separation conditions for two enantiomers of farrerol were determined as follows: Chiralcel OJ-RH(4.6 mm×150 mm, 5 μm), equilibrium elution of acetonitrile-water(40∶60), the flow rate of 0.5 mL·min–1, the column temperature of 25 ℃, and the detection wavelength of 295 nm. Under the optimum separation conditions, the resolution of farrerol enantiomers reached 1.5, indicating that the two enantiomers of the farrerol could be completely separated. When the column temperature was between 20 ℃ and 35 ℃, the separation factor decreased with the increase of temperature. The lnα of the two enantiomers of farrerol showed a good linear relationship with 1/T, and the chiral reselution process was controlled by enthalpy. The enantiomer separation method of farrerol was applied to the determination of farrerol enantiomer in Chinese medicinal materials of Rhododendri Daurici Folium and Rhododendron Micranthum. The linear relationship between the two enantiomers of farrerol were good in the range of 0.718–57.44 μg·mL–1 and 1.28–102.24 μg·mL–1, respectively. And the contents of the two enantiomers of farrerol in Rhododendri Daurici Folium were 0.228 2 and 0.466 2 mg·g–1, respectively. And the contents of the two enantiomers of farrerol in Rhododendron Micranthum were 0.416 8 and 0.707 3 mg·g–1, respectively. CONCLUSION This method is simple, efficient and suitable for the determination of farrerol enantiomers in traditional Chinese medicine.

Objective:To analyze the clinical and pathological data of 15 patients with light chain amyloidosis initially diagnosed with other kidney diseases, and identify possible misdiagnosis reasons.Methods:It was a retrospective observational study. The clinical and pathological data of 15 patients, whose initial kidney biopsies failed to diagnose light chain-amyloidosis but were confirmed by a subsequent kidney biopsy or pathology consultation at Peking University First Hospital from January 2010 to December 2022 were collected. The results of immunofluorescence, Congo red staining, and electron microscopy of two renal biopsies were analyzed.Results:The median age of 15 patients was 56 years old, with a male-to-female ratio of 7∶8. The main clinical manifestation was massive proteinuria with normal kidney function, and there were 10 cases presenting as nephrotic syndrome. The initial diagnosis based on the first kidney biopsy included minimal change disease (8 cases), IgA nephropathy (3 cases), membranous nephropathy (3 cases), and type Ⅲ collagen glomerulonephritis (1 case). M proteinemia was not evaluated in 13 patients during the first kidney biopsy. Light chain immunofluorescence staining was not performed in 12 cases. Congo red staining was not performed in 13 cases. All fifteen patients received glucocorticoids combined with immunosuppressive therapy after their initial diagnosis, and 5 patients developed severe infection. After 12.0 (7.5, 20.0) months of treatment, none of them achieved clinical remission. Thirteen had evidences for M protein before the second kidney biopsy. The renal tissues of all patients underwent immunofluorescence light chain examination, Congo red staining, and immunoelectron microscopy examination when necessary. The repeat kidney biopsies of 14 cases and pathology consultation of one case consistently indicated light chain-amyloidosis. The kidney tissues in 13 cases were confirmed to be light chain restricted, 11 cases by immunofluorescence, and 2 cases by immune electron microscopy. After diagnosis of light chain-amyloidosis, all patients received targeted plasma cell therapy except for 1 patient lost to follow-up, 6 patients achieved hematologic remission, 5 patients achieved renal remission, 1 patient entered end-stage renal disease, and 3 patients died.Conclusions:In middle and elderly-aged patients with nephrotic syndrome, if conventional immunosuppressive therapy yields unsatisfactory results, it is crucial to focus on identifying evidences of monoclonal immunoglobulinemia, if necessary, kidney biopsy should be actively repeated. Kidney biopsy pathology should include comprehensive examinations such as light chain immunofluorescence, Congo red staining, and electron microscopy to avoid misdiagnosis of light chain-amyloidosis.

Objective: To investigate the status and influencing factors of disease uncertainty among caregivers of colorectal cancer patients receiving chemotherapy, so as to provide insights into psychological interventions among caregivers. Methods: Caregivers of colorectal cancer patients hospitalized in Hunan Cancer Hospital, the Third Xiangya Hospital and the Second Xiangya Hospital for chemotherapy from March 2020 to December 2021 were recruited. Caregivers' demographics, health status, medical and nursing support and social support, as well as patients' demographics, frequency of chemotherapy and disease stage were collected using questionnaire surveys. Caregivers' disease uncertainty was evaluated using Chinese version of the Uncertainty in Illness Scale for Family Members, and factors affecting caregivers' disease uncertainty were identified using a multivariable linear regression model. Results: A total of 318 caregivers were enrolled, including 115 men (36.16%) and 203 women (63.84%), with a mean age of (45.89±6.57) years, and there were 186 caregivers as patients' spouses (58.49%). The mean score of disease uncertainty was (86.65±10.82) points, and the scores of the unpredictability dimension, uncertainty dimension, complexity, and lack of information dimension were (3.21±0.35), (2.98±0.48), (2.84±0.49) and (2.36±0.59) points, respectively. There were 285 participants with a high level of disease uncertainty (89.62%), and multivariable linear regression analysis identified social support (β′=-0.199), medical and nursing support (β′=-0.118), patient's age (β′=-0.155) and stage Ⅲ and Ⅳ of colorectal cancer (β′=0.151) as factors affecting caregiver's disease uncertainty. Conclusion Caregivers of colorectal cancer patients with chemotherapy have a high level of disease uncertainty, which is affected by social support, medical and nursing support, patient's age and duration of disease.

The paper reports a case of coenzyme Q10 deficiency nephrotic syndrome associated with coenzyme Q2 gene mutation and reviews the literature on this topic. The patient presented with hematuria, proteinuria, and a diminution of vision as clinical manifestations. But the proteinuria was not relieved after sufficient doses of glucocorticoids for over 2 months. The patient′s birth history was unremarkable, and his parents were both healthy and not consanguineous. Whole exome sequencing revealed that the patient had a mutation of coenzyme Q2 gene at c.973A>G(p.T325A) and c.517C>T(p.R173C). Combined with renal biopsy pathology, the patient was diagnosed with hereditary nephropathy and started the supplements of coenzyme Q10 after stopping glucocorticoid treatments immediately. After 5 weeks of therapy, the patient′s 24-h urine protein quantification decreased from 6.01 g to 1.53 g.

Objective:To investigate the urinary sediment findings and the clinicopathologic features of IgA nephropathy (IgAN) patients with acute kidney injury (AKI).Methods:It was a retrospective study. The patients with renal biopsy-proven primary IgAN in Peking University First Hospital from January 31, 2013 to July 31, 2015 were selected. According to whether AKI occurred at renal biopsy or not, the patients were divided into AKI group and non-AKI group. Morning urine samples were obtained on the day of renal biopsy. Urine sediments, including various cells and casts, were examined. The clinical data, urinary sediments, and renal pathological changes were compared between the two groups. Logistic regression analysis was performed to identify the association between clinical pathological changes, urinary sediment indicators and AKI, or clinical pathological changes and urinary sediment indicators.Results:There were 502 IgAN patients enrolled in this study, with age of (36.1±12.1) years old and 261 males (52.0%). The incidence of AKI was 11.4% (57/502) among the enrolled patients at the time of renal biopsy. Common causes of AKI included gross hematuria-induced AKI (10 cases), acute tubulointerstitial nephritis (10 cases), crescentic IgAN (9 cases), malignant hypertensive renal damage (6 cases), and multiple etioloqy or unknown etiology (22 cases). Compared with non-AKI group, AKI group had higher proportions of males and malignant hypertension, higher levels of proteinuria and urinary erythrocyte counts, and higher frequencies of gross hematuria, leukocyturia, renal tubular epithelial cells, and granular casts (all P<0.05). AKI group also had higher proportions of severe tubular atrophy/interstitial fibrosis (T2) and cellular/cellular fibrous crescent formation (C2) than non-AKI group (both P<0.05). Logistic regression analysis results showed that, there were statistically significant differences in the correlation between AKI and gender, 24 h urinary protein, urinary erythrocyte counts, granular casts and renal tubular atrophy/interstitial fibrosis (T) scores (all P<0.05). Hematuria, leukocyturia, red blood cell casts, white blood cell casts, granular casts, and fatty casts were correlated with endothelial hypercellularity (E) and cellular/cellular fibrous crescent formation (C) scores, respectively (all P<0.05). Hematuria was correlated with mesangial hypercellularity (M) scores ( OR=2.613, 95% CI 1.520-4.493, P=0.001). Hematuria ( OR=1.723, 95% CI 1.017-2.919, P=0.043) and fatty casts ( OR=2.646, 95% CI 1.122-6.238, P=0.026) were correlated with segmental sclerosis or adhesion (S) scores. Leukocyturia ( OR=1.645, 95% CI 1.154-2.347, P=0.006) and fatty casts ( OR=2.344, 95% CI 1.202-4.572, P=0.012) were correlated with T scores. Epithelial cell cast was correlated with C scores ( OR=1.857, 95% CI 1.174-2.939, P=0.008). Conclusions:AKI is a common complication among IgAN patients with diverse etiology and more severe clinicopathological features. Urinary sediment findings can reflect renal pathological changes to some extent, and therefore assist in the clinical diagnosis and treatment of IgAN patients with AKI.

Objective:To understand the status quo of emotional intelligence, emotional labor, and humanistic caring ability of medical staff, and to clarify their internal relationship.Methods:Convenience sampling was used to select 713 medical staff from a grade A tertiary hospital in Chengdu, Sichuan Province, China. Emotional Intelligence Scale, Humanistic Caring Scale, and Emotional Labor Scale were used to measure the emotional intelligence, humanistic caring ability, and emotional labor of medical staff. SPSS 22.0 software was used to establish a database for statistical description and analysis. Process 3.2 software was used to analyze the mediating effect.Results:In humanistic caring ability, the average score of comprehension dimension was the highest (75.62±8.20) and the average score of patience dimension was the lowest (58.53±5.01). In emotional labor, the average score of the deep action dimension was the highest (23.39±3.85) and the average score of the surface action dimension was the lowest (17.73±3.18). In emotional intelligence, the average score of self-emotion evaluation dimension was the highest (21.76±3.30) and the average score of other-emotion evaluation dimension was the lowest (20.07±3.71). Positive correlations were found between humanistic caring ability and emotional intelligence, between humanistic caring ability and emotional labor, and between emotional intelligence and emotional labor ( P<0.01). Humanistic caring ability had a partial mediating effect between emotional intelligence and emotional labor. Humanistic caring ability had direct and indirect effects on emotional labor, and the effect sizes were 0.279 and 0.029, respectively. Conclusion:Emotional intelligence has a direct positive predictive effect on emotional labor, humanistic caring ability as an intermediary variable indirectly and positively predicts emotional labor. Humanistic caring ability plays a partial mediating role between emotional intelligence and emotional labor. Attention should be paid to the emotional labor of medical staff, and the emotional labor of medical staff should be improved through targeted training on emotional intelligence and humanistic caring ability. These efforts will improve the current situation and establish a harmonious doctor-patient relationship.

Purpose: The role of vacuolar protein sorting 34 (Vps34), an indispensable protein required for cell vesicular trafficking, in the biological behavior of hepatocellular carcinoma (HCC) has yet to be studied. Materials and Methods: In the present study, the expression of Vps34 in HCC and the effect of Vps34 on HCC cell invasion was detected both in vivo and in vitro. Furthermore, by modulating the RILP and Rab11, which regulate juxtanuclear lysosome aggregation and recycling endosome respectively, the underlying mechanism was investigated. Results: Vps34 was significantly decreased in HCC and negatively correlated with the HCC invasiveness both in vivo and in vitro. Moreover, Vps34 could promote lysosomal juxtanuclear accumulation, reduce the invasive ability of HCC cells via the Rab7-RILP pathway. In addition, the deficiency of Vps34 in HCC cells affected the endosome-lysosome system, resulting in enhanced Rab11 mediated endocytic recycling of cell surface receptor and increased invasion of HCC cells. Conclusion Our study reveals that Vps34 acts as an invasion suppressor in HCC cells, and more importantly, the endosome-lysosome trafficking regulated by Vps34 has the potential to become a target pathway in HCC treatment.

Pancreatic ductal adenocarcinoma (PDAC) is characterized by the highest mortality among carcinomas. The pathogenesis of PDAC requires elevated autophagy, inhibition of which using hydroxychloroquine has shown promise. However, current realization is impeded by its suboptimal use and unpredictable toxicity. Attempts to identify novel autophagy-modulating agents from already approved drugs offer a rapid and accessible approach. Here, using a patient-derived organoid model, we performed a comparative analysis of therapeutic responses among various antimalarial/fungal/parasitic/viral agents, through which econazole (ECON), an antifungal compound, emerged as the top candidate. Further testing in cell-line and xenograft models of PDAC validated this activity, which occurred as a direct consequence of dysfunctional autophagy. More specifically, ECON boosted autophagy initiation but blocked lysosome biogenesis. RNA sequencing analysis revealed that this autophagic induction was largely attributed to the altered expression of activation transcription factor 3 (ATF3). Increased nuclear import of ATF3 and its transcriptional repression of inhibitor of differentiation-1 (ID-1) led to inactivation of the AKT/mammalian target of rapamycin (mTOR) pathway, thus giving rise to autophagosome accumulation in PDAC cells. The magnitude of the increase in autophagosomes was sufficient to elicit ER stress-mediated apoptosis. Furthermore, ECON, as an autophagy inhibitor, exhibited synergistic effects with trametinib on PDAC. This study provides direct preclinical and experimental evidence for the therapeutic efficacy of ECON in PDAC treatment and reveals a mechanism whereby ECON inhibits PDAC growth.

Objective:To analyze clinical data of 3 children with LMX1B-associated disease characterized by asymptomatic glomerular proteinuria, thus improving the recognition of asymptomatic proteinuria with genetic causes. Methods:Three patients with LMX1B-associated disease presented with prominent asymptomatic proteinuria diagnosed by the next-generation sequencing in Department of Pediatrics, Peking University First Hospital from April 2014 to October 2017 were included in this study.Clinical data, including renal and extrarenal manifestations, renal biopsy, and family history, were collected and retrospectively analyzed. Results:All 3 children were girls, the age of onset were 2 years, 1 year, and 4 years, respectively, and the diagnosis age were 11 years, 5 years and 6 years, respectively.All of them had glomerular proteinuria, and nephrotic-level proteinuria occurred in one patient.Microscopic hematuria was found in 2 patients.All of them had normal renal function.Only one patient underwent renal biopsy.Electron microscopy of the first time of biopsy revealed segmental thinning of the glomerular basement membrane.Re-biopsy 4 years later showed irregular thickening of the glomerular basement membrane, moth-eaten appearance and collagen fibrillar material deposition.No abnormalities of nails, limbs and joints were observed by physical examination.Two patients had a family history of renal disease.Conclusions:Genetic factors should be considered in children with obscure onset asymptomatic proteinuria without definite clinical causes.Genetic testing can help diagnose and guide treatment as early as possible.

GB7 acetate is a galbulimima alkaloid obtained from Galbulimima belgraveana.However,information regarding its structure,biological activities,and related mechanisms is not entirely available.A series of spectroscopic analyses,structural degradation,interconversion,and crystallography were performed to identify the structure of GB7 acetate.The MTT assay was applied to measure cell proliferation on human colorectal cancer HCT 116 cells.The expressions of the related proteins were measured by Western blotting.Transmission electron microscopy(TEM),acridine orange(AO)and monodansylcadaverine(MDC)staining were used to detect the presence of autophagic vesicles and autolysosomes.A transwell assay was performed to demonstrate metastatic capabilities.Oxygen consumption rate(OCR)and extracellular acidification rate(ECAR)assays were performed to determine the mitochondrial oxidative phosphorylation(OXPHOS)and glycolysis activity of HCT 116 cells.The data showed that GB7 acetate suppressed the proliferation and colony-forming ability of HCT 116 cells.Pretreatment with GB7 acetate significantly induced the formation of autophagic vesicles and autolysosomes.GB7 acetate upregulated the expressions of LC3 and Thr172 phosphorylated adenosine 5'-monophosphate(AMP)-activated pro-tein kinase α(p-AMPKα),which are key elements of autophagy.In addition,GB7 acetate suppressed the metastatic capabilities of HCT 116 cells.Additionally,the production of matrix metallo-proteinase-2(MMP-2)and MMP-9 was reduced,whereas the expression of E-cadherin(E-cad)was upregulated.Furthermore,GB7 acetate significantly reduced mitochondrial OXPHOS and glycolysis.In conclusion,the structure of the novel Galbulimima alkaloid GB7 acetate was identified.GB7 acetate was shown to have anti-proliferative,pro-autophagic,anti-metastatic,and anti-metabolite capabilities in HCT 116 cells.This study might provide new insights into cancer treatment efficacy and cancer chemoprevention.