OBJECTIVE: To investigate the effects of astragaloside IV (AS-IV) on podocyte injury of diabetic nephropathy (DN) and reveal its potential mechanism. METHODS: In in vitro experiment, podocytes were divided into 4 groups, normal, high glucose (HG), inositol-requiring enzyme 1 (IRE-1) α activator (HG+thapsigargin 1 µmol/L), and IRE-1α inhibitor (HG+STF-083010, 20 µmol/L) groups. Additionally, podocytes were divided into 4 groups, including normal, HG, AS-IV (HG+AS-IV 20 µmol/L), and IRE-1α inhibitor (HG+STF-083010, 20 µmol/L) groups, respectively. After 24 h treatment, the morphology of podocytes and endoplasmic reticulum (ER) was observed by electron microscopy. The expressions of glucose-regulated protein 78 (GRP78) and IRE-1α were detected by cellular immunofluorescence. In in vivo experiment, DN rat model was established via a consecutive 3-day intraperitoneal streptozotocin (STZ) injections. A total of 40 rats were assigned into the normal, DN, AS-IV [AS-IV 40 mg/(kg·d)], and IRE-1α inhibitor [STF-083010, 10 mg/(kg·d)] groups (n=10), respectively. The general condition, 24-h urine volume, random blood glucose, urinary protein excretion rate (UAER), urea nitrogen (BUN), and serum creatinine (SCr) levels of rats were measured after 8 weeks of intervention. Pathological changes in the renal tissue were observed by hematoxylin and eosin (HE) staining. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect the expressions of GRP78, IRE-1α, nuclear factor kappa Bp65 (NF-κBp65), interleukin (IL)-1β, NLR family pyrin domain containing 3 (NLRP3), caspase-1, gasdermin D-N (GSDMD-N), and nephrin at the mRNA and protein levels in vivo and in vitro, respectively. RESULTS: Cytoplasmic vacuolation and ER swelling were observed in the HG and IRE-1α activator groups. Podocyte morphology and ER expansion were improved in AS-IV and IRE-1α inhibitor groups compared with HG group. Cellular immunofluorescence showed that compared with the normal group, the fluorescence intensity of GRP78 and IRE-1α in the HG and IRE-1α activator groups were significantly increased whereas decreased in AS-IV and IRE-1α inhibitor groups (P<0.05). Compared with the normal group, the mRNA and protein expressions of GRP78, IRE-1α, NF-κ Bp65, IL-1β, NLRP3, caspase-1 and GSDMD-N in the HG group was increased (P<0.05). Compared with HG group, the expression of above indices was decreased in the AS-IV and IRE-1α inhibitor groups, and the expression in the IRE-1α activator group was increased (P<0.05). The expression of nephrin was decreased in the HG group, and increased in AS-IV and IRE-1α inhibitor groups (P<0.05). The in vivo experiment results revealed that compared to the normal group, the levels of blood glucose, triglyceride, total cholesterol, BUN, blood creatinine and urinary protein in the DN group were higher (P<0.05). Compared with DN group, the above indices in AS-IV and IRE-1α inhibitor groups were decreased (P<0.05). HE staining revealed glomerular hypertrophy, mesangial widening and mesangial cell proliferation in the renal tissue of the DN group. Compared with the DN group, the above pathological changes in renal tissue of AS-IV and IRE-1α inhibitor groups were alleviated. Quantitative RT-PCR and Western blot results of GRP78, IRE-1α, NF-κ Bp65, IL-1β, NLRP3, caspase-1 and GSDMD-N were consistent with immunofluorescence analysis. CONCLUSION AS-IV could reduce ERS and inflammation, improve podocyte pyroptosis, thus exerting a podocyte-protective effect in DN, through regulating IRE-1α/NF-κ B/NLRP3 signaling pathway.
Podocytes/metabolism* ; Animals ; Diabetic Nephropathies/metabolism* ; Saponins/therapeutic use* ; Triterpenes/therapeutic use* ; Signal Transduction/drug effects* ; NF-kappa B/metabolism* ; Protein Serine-Threonine Kinases/metabolism* ; Male ; Rats, Sprague-Dawley ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Endoribonucleases/metabolism* ; Endoplasmic Reticulum Chaperone BiP ; Rats ; Diabetes Mellitus, Experimental/complications* ; Endoplasmic Reticulum/metabolism* ; Multienzyme Complexes

OBJECTIVE: To explore the mechanism of colon dialysis with Yishen Decoction (YS) in improving the autophagy disorder of intestinal epithelial cells in chronic renal failure (CRF) in vivo and in vitro. METHODS: Thirty male SD rats were randomly divided into normal, CRF, and colonic dialysis with YS groups by a random number table method (n=10). The CRF model was established by orally gavage of adenine 200 mg/(kg•d) for 4 weeks. CRF rats in the YS group were treated with colonic dialysis using YS 20 g/(kg•d) for 14 consecutive days. The serum creatinine (SCr) and urea nitrogen (BUN) levels were detected by enzyme-linked immunosorbent assay. Pathological changes of kidney and colon tissues were observed by hematoxylin and eosin staining. Autophagosome changes in colonic epithelial cells was observed with electron microscopy. In vitro experiments, human colon cancer epithelial cells (T84) were cultured and divided into normal, urea model (74U), YS colon dialysis, autophagy activator rapamycin (Ra), autophagy inhibitor 3-methyladenine (3-MA), and SIRT1 activator resveratrol (Re) groups. RT-PCR and Western blot were used to detect the mRNA and protein expressions of zonula occludens-1 (ZO-1), Claudin-1, silent information regulator sirtuin 1 (SIRT1), LC3, and Beclin-1 both in vitro and in vivo. RESULTS: Colonic dialysis with YS decreased SCr and BUN levels in CRF rats (P<0.05), and alleviated the pathological changes of renal and colon tissues. Expressions of SIRT1, ZO-1, Claudin-1, Beclin-1, and LC3II/I were increased in the YS group compared with the CRF group in vivo (P<0.05). In in vitro study, compared with normal group, the expressions of SIRT1, ZO-1, and Claudin-1 were decreased, and expressions of Beclin-1, and LC3II/I were increased in the 74U group (P<0.05). Compared with the 74U group, expressions of SIRT1, ZO-1, and Claudin-1 were increased, whereas Beclin-1, and LC3II/I were decreased in the YS group (P<0.05). The treatment of 3-MA and rapamycin regulated autophagy and the expression of SIRT1. SIRT1 activator intervention up-regulated autophagy as well as the expressions of ZO-1 and Claudin-1 compared with the 74U group (P<0.05). CONCLUSION Colonic dialysis with YS could improve autophagy disorder and repair CRF intestinal mucosal barrier injury by regulating SIRT1 expression in intestinal epithelial cells.
Animals ; Sirtuin 1/metabolism* ; Drugs, Chinese Herbal/therapeutic use* ; Autophagy/drug effects* ; Male ; Intestinal Mucosa/drug effects* ; Rats, Sprague-Dawley ; Epithelial Cells/metabolism* ; Colon/drug effects* ; Humans ; Kidney Failure, Chronic/drug therapy* ; Signal Transduction/drug effects* ; Renal Dialysis ; Rats ; Kidney/drug effects*

Objective To construct and validate a risk prediction model for in-hospital death af-ter successful resuscitation in patients with cardiac arrest.Methods A retrospective study was con-ducted on 295 patients with cardiac arrest who successfully restored spontaneous circulation after car-diopulmonary resuscitation and were further treated in hospital.The patients were divided into training and validation sets using K-fold cross-validation and then grouped and compared based on whether in-hospital death occurred.A binary Logistic regression analysis was used to screen risk prediction fac-tors,and a nomogram prediction model was constructed.The model performance was evaluated and validated in the training and validation sets,respectively.Results The results of the multivariate Logistic regression analysis showed that hospitalization duration(OR=1.180;95％CI,1.080 to 1.280;P＜0.001),norepinephrine dose(OR=0.980;95％CI,0.970 to 0.990;P=0.002),ini-tial respiratory rate after resuscitation(OR=1.090;95％CI,1.030 to 1.150;P=0.004),and sinus rhythm recovery after resuscitation(OR=4.280;95％CI,1.670 to 10.980;P=0.003)were inde-pendent influencing factors for in-hospital death.A nomogram model was constructed based on these in-dependent influencing factors,and it was verified that the model had good discrimination,calibration,applicability,and rationality.Conclusion The influencing factors for in-hospital death after successful resuscitation in patients with cardiac arrest include hospitalization duration,norepinephrine dose,initial respiratory rate after resuscitation,and sinus rhythm recovery after resuscitation.The nomo-gram model constructed based on these factors can provide a reference for clinical decision-making.

OBJECTIVE: Exposure to polycyclic aromatic hydrocarbons (PAHs) or metal(loid)s individually has been associated with neural tube defects (NTDs). However, the impacts of PAH and metal(loid) co-exposure and potential interaction effects on NTD risk remain unclear. We conducted a case-control study in China among population with a high prevalence of NTDs to investigate the combined effects of PAH and metal(loid) exposures on the risk of NTD. METHODS: Cases included 80 women who gave birth to offspring with NTDs, whereas controls were 50 women who delivered infants with no congenital malformations. We analyzed the levels of placental PAHs using gas chromatography and mass spectrometry, PAH-DNA adducts with ³²P-post-labeling method, and metal(loid)s with an inductively coupled plasma mass spectrometer. Unconditional logistic regression was employed to estimate the associations between individual exposures and NTDs. Least absolute shrinkage and selection operator (LASSO) penalized regression models were used to select a subset of exposures, while additive interaction models were used to identify interaction effects. RESULTS: In the single-exposure models, we found that eight PAHs, PAH-DNA adducts, and 28 metal(loid)s were associated with NTDs. Pyrene, selenium, molybdenum, cadmium, uranium, and rubidium were selected through LASSO regression and were statistically associated with NTDs in the multiple-exposure models. Women with high levels of pyrene and molybdenum or pyrene and selenium exhibited significantly increased risk of having offspring with NTDs, indicating that these combinations may have synergistic effects on the risk of NTDs. CONCLUSION Our findings suggest that individual PAHs and metal(loid)s, as well as their interactions, may be associated with the risk of NTDs, which warrants further investigation.
Humans ; Neural Tube Defects/chemically induced* ; Polycyclic Aromatic Hydrocarbons/adverse effects* ; Female ; Case-Control Studies ; China/epidemiology* ; Adult ; Pregnancy ; Environmental Pollutants ; Maternal Exposure/adverse effects* ; Metals/toxicity* ; Young Adult ; Risk Factors

Objective:To investigate GPRC5D expression on myeloma cells in newly diagnosed multiple myeloma (NDMM) patients and evaluate its prognostic significance.Methods:This study retrospectively analyzed the clinical data of 65 patients with NDMM treated at the Affiliated Hospital of Xuzhou Medical University from April 2023 to April 2024. The expression of GPRC5D on the surface of myeloma cells in all patients was detected with flow cytometry before induction therapy, and patients were stratified into high and low GPRC5D expression groups based on the median GPRC5D positivity rate. Clinical characteristics, immune status, treatment response after induction therapy, and prognosis were compared between the two groups.Results:The median positive rate of GPRC5D in the plasma cells of 65 patients with NDMM was 32.68%. Based on this threshold, patients were categorized into the high (33 cases, GPRC5D positive rate ≥ 32.68%) and low (32 cases, GPRC5D positive rate <32.68%) GPRC5D expression groups. Compared with the low GPRC5D expression group, the high GPRC5D expression group demonstrated a higher proportion of 1q21 gain (78.8% vs 43.8%, P=0.004), a higher incidence of immunoparesis involving ≥2 uninvolved immunoglobulins (87.9% vs 62.5%, P=0.018), and severe immunoparesis (59.4% vs 33.3%, P=0.046). Further, CD16 +CD56 + cell levels were lower in the high GPRC5D expression group [ (16.60±8.70) % vs (27.78±15.78) %, P=0.005]. No significant difference was observed in the overall response rate between the high and low GPRC5D expression groups (78.8% vs 93.8%, P=0.165). However, the high GPRC5D expression group exhibited a significantly lower rate of achieving very good partial remission or better (42.4% vs 78.2%, P=0.003) and a lower MRD negativity rate (30.0% vs 68.8%, P=0.002). Compared with the low GPRC5D expression group, patients with high expression demonstrated a significantly shorter median progression-free survival (11.2 months vs not reached, P=0.002), whereas the median overall survival was not reached in either group, with no statistically significant difference ( P=0.069) . Conclusions:The GPRC5D positivity rate in the plasma cells of patients with NDMM is associated with 1q21 gain and immune status. High GPRC5D expression at diagnosis may predict poor response to induction therapy and an unfavorable prognosis.

Objective:To investigate impact of farrerol(Far)on LPS-induced microglial inflammatory injury by regulating cyclic guanosine monophosphate-adenosine monophosphate synthase(cGAS)-stimulator of interferon genes(STING)signal pathway.Methods:Mice BV2 microglial cell lines were grouped into control group(normal culture),LPS group(1 μg/ml),Far low,medium and high doses groups(1,5,10 μmol/L),activator group(10 μmol/L Far+75 μg/ml cGAS-STING signal pathway activator DMX-AA);proliferation and apoptosis of BV2 cells were detected by MTT,plate cloning assay and flow cytometry;qRT-PCR and ELISA were applied to detect levels of IL-6,IL-1β and TNF-α in cells and supernatants;NO content in cell supernatant was detected by nitrate reductase method;Western blot was applied to detect expressions of proliferating cell nuclear antigen(PCNA),Bcl-2 associated X protein(Bax),anti-apoptotic factor B cell lymphoma protein-2(Bcl-2)and cGAS-STING pathway protein in BV2 cells.Results:Compared with control group,A490 of BV2 cells,number of cloned cells,expressions of PCNA and Bcl-2 proteins in LPS group were decreased,apoptosis rate,mRNA expressions of IL-6,IL-1β,TNF-α,contents of IL-6,IL-1β,TNF-α,NO,and protein expres-sions of Bax,cGAS and STING were increased(P<0.05);compared with LPS group,A490 of BV2 cells,number of cloned cells,expressions of PCNA and Bcl-2 proteins in Far low,medium and high dose groups were increased,apoptosis rate,mRNA expressions of IL-6, IL-1β, TNF-α, contents of IL-6, IL-1β, TNF-α, NO, and protein expressions of Bax, cGAS and STING were decreased (P<0.05); compared with Far high-dose group, A490 of BV2 cells, number of cloned cells, expressions of PCNA and Bcl-2 proteins in activator group were decreased, apoptosis rate, mRNA expressions of IL-6, IL-1β, TNF-α, contents of IL-6, IL-1β, TNF-α, NO, and protein expressions of Bax, cGAS and STING were increased (P<0.05).Conclusion:Far may inhibit apoptosis and inflammatory injury of BV2 cells and promote proliferation of BV2 cells by inhibiting cGAS-STING pathway, and thus alleviate inflammatory injury of BV2 cells induced by LPS.

Objective:To investigate impact of farrerol(Far)on LPS-induced microglial inflammatory injury by regulating cyclic guanosine monophosphate-adenosine monophosphate synthase(cGAS)-stimulator of interferon genes(STING)signal pathway.Methods:Mice BV2 microglial cell lines were grouped into control group(normal culture),LPS group(1 μg/ml),Far low,medium and high doses groups(1,5,10 μmol/L),activator group(10 μmol/L Far+75 μg/ml cGAS-STING signal pathway activator DMX-AA);proliferation and apoptosis of BV2 cells were detected by MTT,plate cloning assay and flow cytometry;qRT-PCR and ELISA were applied to detect levels of IL-6,IL-1β and TNF-α in cells and supernatants;NO content in cell supernatant was detected by nitrate reductase method;Western blot was applied to detect expressions of proliferating cell nuclear antigen(PCNA),Bcl-2 associated X protein(Bax),anti-apoptotic factor B cell lymphoma protein-2(Bcl-2)and cGAS-STING pathway protein in BV2 cells.Results:Compared with control group,A490 of BV2 cells,number of cloned cells,expressions of PCNA and Bcl-2 proteins in LPS group were decreased,apoptosis rate,mRNA expressions of IL-6,IL-1β,TNF-α,contents of IL-6,IL-1β,TNF-α,NO,and protein expres-sions of Bax,cGAS and STING were increased(P<0.05);compared with LPS group,A490 of BV2 cells,number of cloned cells,expressions of PCNA and Bcl-2 proteins in Far low,medium and high dose groups were increased,apoptosis rate,mRNA expressions of IL-6, IL-1β, TNF-α, contents of IL-6, IL-1β, TNF-α, NO, and protein expressions of Bax, cGAS and STING were decreased (P<0.05); compared with Far high-dose group, A490 of BV2 cells, number of cloned cells, expressions of PCNA and Bcl-2 proteins in activator group were decreased, apoptosis rate, mRNA expressions of IL-6, IL-1β, TNF-α, contents of IL-6, IL-1β, TNF-α, NO, and protein expressions of Bax, cGAS and STING were increased (P<0.05).Conclusion:Far may inhibit apoptosis and inflammatory injury of BV2 cells and promote proliferation of BV2 cells by inhibiting cGAS-STING pathway, and thus alleviate inflammatory injury of BV2 cells induced by LPS.

Objective:To investigate GPRC5D expression on myeloma cells in newly diagnosed multiple myeloma (NDMM) patients and evaluate its prognostic significance.Methods:This study retrospectively analyzed the clinical data of 65 patients with NDMM treated at the Affiliated Hospital of Xuzhou Medical University from April 2023 to April 2024. The expression of GPRC5D on the surface of myeloma cells in all patients was detected with flow cytometry before induction therapy, and patients were stratified into high and low GPRC5D expression groups based on the median GPRC5D positivity rate. Clinical characteristics, immune status, treatment response after induction therapy, and prognosis were compared between the two groups.Results:The median positive rate of GPRC5D in the plasma cells of 65 patients with NDMM was 32.68%. Based on this threshold, patients were categorized into the high (33 cases, GPRC5D positive rate ≥ 32.68%) and low (32 cases, GPRC5D positive rate <32.68%) GPRC5D expression groups. Compared with the low GPRC5D expression group, the high GPRC5D expression group demonstrated a higher proportion of 1q21 gain (78.8% vs 43.8%, P=0.004), a higher incidence of immunoparesis involving ≥2 uninvolved immunoglobulins (87.9% vs 62.5%, P=0.018), and severe immunoparesis (59.4% vs 33.3%, P=0.046). Further, CD16 +CD56 + cell levels were lower in the high GPRC5D expression group [ (16.60±8.70) % vs (27.78±15.78) %, P=0.005]. No significant difference was observed in the overall response rate between the high and low GPRC5D expression groups (78.8% vs 93.8%, P=0.165). However, the high GPRC5D expression group exhibited a significantly lower rate of achieving very good partial remission or better (42.4% vs 78.2%, P=0.003) and a lower MRD negativity rate (30.0% vs 68.8%, P=0.002). Compared with the low GPRC5D expression group, patients with high expression demonstrated a significantly shorter median progression-free survival (11.2 months vs not reached, P=0.002), whereas the median overall survival was not reached in either group, with no statistically significant difference ( P=0.069) . Conclusions:The GPRC5D positivity rate in the plasma cells of patients with NDMM is associated with 1q21 gain and immune status. High GPRC5D expression at diagnosis may predict poor response to induction therapy and an unfavorable prognosis.

AIM To study the phenylpropanoids from Brandisia hancei Hook.f.and their antioxidant activities.METHODS The extract from B.hancei was isolated and purified by Rp-C18,MCI,semi-preparative HPLC,silica gel and Sephadex LH-20,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The cytotoxicities was determined by MTT method,and the antioxidant activities were determined by DPPH and ABTS+free radical scavenging methods.RESULTS Fifteen phenylpropanoids were isolated and identified as(+)-pinonesinol(1),(-)-medioresinol(2),(-)-syringaresinol(3),buddlenol D(4),(7R,7'R,7″S,8S,8'S,8″S)-4',5″-dihydroxy-3,5,3',4″-tetramethoxy-7,9':7',9-diepoxy-4,8″-oxy-8,8'-sesquineo-lignan-7″,9″-diol(5),(-)-(7R,7'R,7″R,8S,8'S,8″S)-4',4″-dihydroxy-3,3',3″,5-tetramethoxy-7,9':7',9-diepoxy-4,8″-oxy-8,8'-sesquineolignan-7″,9″-diol(6),hedyotol A(7),dracunculifoside R(8),acteoside(9),isoacteoside(10),arenarioside(11),isomartynoside(12),curcasinlignan B(13),erythro-2,3-bis(4-hydroxy-3-methoxyphenyl)-3-ethoxypropan-l-ol(14),citrusin C(15).Compounds 1-4 and 9-10 had no obvious cytotoxicity to HepG2 hepatoma cells.Compounds 1,3,9,10 and 12 had strong scavenging activities against DPPH radicals.Compounds 1-3,9-10,12 and 14 showed strong scavenging activities against ABTS+radical.CONCLUSION Compounds 1-8 and 12-15 are isolated from genus Brandisia for the first time.The phenylpropanoids from B.hancei show strong antioxidant activities.

AIM To study the chemical constituents from the stems of Gnetum parvifolium(Warb.)C.Y.Cheng ex Chun and their xanthine oxidase inhibitory activities.METHODS The 95%ethanol extract from the stems of G.parvifolium was isolated and purified by silica gel,MCI and preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The inhibitory activities on xanthine oxidase was determined by ultraviolet spectrophotometry.RESULTS Seventeen compounds were isolated and identified as isorhapontigenin(1),resveratrol(2),4-hydroxybenzaldehyde(3),cycloeucalenol(4),(E)-1-(3',5'-dimethoxyphenyl)-2-(3,4-dimethoxyphenyl)ethene(5),stigmast-4-en-3-one(6),medioresinol(7),chrysin(8),aurantiamide acetate(9),(-)-pinoresinol(10),methyl 4-hydroxybenzoate(11),2-methoxybenzene-1,4-diol(12),gnetumontain C(13),syringaresinol(14),homoeriodictyol(15),vanillin(16),β-sitosterol(17).The IC50 values of compounds 1-3,15-16 were(11.4±0.54),(14.1±1.06),(320.4±0.75),(360.6±0.78),(386.3±0.71)μmol/L,respectively.CONCLUSION Compounds 3-4 are isolated from this plant for the first time.Compounds 5-13 are first isolated from genus Gnetum.Compounds 1-3,15-16 have xanthine oxidase inhibitory activities.