ObjectiveThis study aims to investigate the effect of Dictamni Cortex on intestinal barrier damage in rats and its mechanism by untargeted metabolomics and targeted metabolomics for short-chain fatty acids （SCFAs）. MethodsRats were randomly divided into a control group， a high-dose group of Dictamni Cortex （8.1 g·kg^-1）， a medium-dose group （2.7 g·kg^-1）， and a low-dose group （0.9 g·kg^-1）. Except for the control group， the other groups were administered different doses of Dictamni Cortex by gavage for eight consecutive weeks. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in the ileal tissue. Enzyme-linked immunosorbent assay （ELISA） was employed to detect the level of cytokines， including tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β）， in the ileal tissue of rats. Quantitative real-time fluorescence polymerase chain reaction （Real-time PCR） technology was used to detect the expression level of tight junction proteins， including zonula occludens-1 （ZO-1）， Occludin， and Claudin-1 mRNAs， in the ileal tissue of rats to preliminarily explore the effects of Dictamni Cortex on intestinal damage. The dose with the most significant toxic phenotype was selected to further reveal the effects of Dictamni Cortex on the metabolic profile of ileal tissue in rats by non-targeted metabolomics combined with targeted metabolomics for SCFAs. ResultsCompared with the control group， all doses of Dictamni Cortex induced varying degrees of pathological damage in the ileum， increased TNF-α （P<0.01）， IL-6 （P<0.01）， and IL-1β （P<0.01） levels in the ileal tissue， and decreased the expression level of ZO-1 （P<0.05， P<0.01）， Occludin （P<0.01）， and Claudin-1 （P<0.05） in the ileal tissue， with the high-dose group showing the most significant toxic phenotypes. The damage mechanisms of the high-dose group of Dictamni Cortex on the ileal tissue were further explored by integrating non-targeted metabolomics and targeted metabolomics for SCFAs. The non-targeted metabolomics results showed that 21 differential metabolites were identified in the control group and the high-dose group. Compared with that in the control group， after Dictamni Cortex intervention， the level of 14 metabolites was significantly increased （P<0.05， P<0.01）， and the level of seven metabolites was significantly decreased （P<0.05， P<0.01） in the ileal contents. These metabolites collectively acted on 10 related metabolic pathways， including glycerophospholipids and primary bile acid biosynthesis. The quantitative data of targeted metabolomics for SCFAs showed that Dictamni Cortex intervention disrupted the level of propionic acid， butyric acid， acetic acid， caproic acid， isobutyric acid， isovaleric acid， valeric acid， and isocaproic acid in the ileal contents of rats. Compared with those in the control group， the level of isobutyric acid， isovaleric acid， and valeric acid were significantly increased， while the level of propionic acid， butyric acid， and acetic acid were significantly decreased in the ileal contents of rats after Dictamni Cortex intervention （P<0.05， P<0.01）. ConclusionDictamni Cortex can induce intestinal damage by regulating glycerophospholipid metabolism， primary bile acid biosynthesis， and metabolic pathways for SCFAs.

Objective To investigate whether sodium butyrate(NaB)and sorafenib synergistically induces ferroptosis to suppress proliferation of hepatocellular carcinoma cells and the possible underlying mechanisms.Methods CCK8 assay and colony formation assay were used to assess the effects of NaB and sorafenib,alone or in combination,on proliferation of HepG2 cells,and ferroptosis of the treated cells was detected with GSH assay and C11-BODIPY 581/591 fluorescent probe.TCGA database was used to analyze differential YAP gene expression between liver cancer and normal tissues.The effects of NaB and sorafenib on YAP and p-YAP expressions in HepG2 cells were invesitigated using Western blotting.Results NaB(2 mmol/L)significantly reduced the IC50 of sorafenib in HepG2 cells,and combination index analysis confirmed the synergy between sorafenib and NaB.The ferroptosis inhibitor Fer-1 and the YAP activator(XMU)obviously reversed the growth-inhibitory effects of the combined treatment with NaB and sorafenib in HepG2 cells.The combined treatment with NaB and sorafenib,as compared with the two agents used alone,significantly inhibited colony formation of HepG2 cells,further enhanced cellular shrinkage and dispersion,and decreased intracellular GSH and lipid ROS levels,and these effects were reversed by Fer-1 and XMU.TCGA analysis revealed a higher YAP mRNA expression in liver cancer tissues than in normal liver tissues.NaB combined with sorafenib produced significantly stronger effects than the individual agents for downregulating YAP protein expression and upregulating YAP phosphorylation level in HepG2 cells.Conclusion NaB combined with sorafenib synergistically inhibit hepatocellular carcinoma cell proliferation possibly by inducing ferroptosis via inhibiting YAP expression.

Objective To investigate whether sodium butyrate(NaB)and sorafenib synergistically induces ferroptosis to suppress proliferation of hepatocellular carcinoma cells and the possible underlying mechanisms.Methods CCK8 assay and colony formation assay were used to assess the effects of NaB and sorafenib,alone or in combination,on proliferation of HepG2 cells,and ferroptosis of the treated cells was detected with GSH assay and C11-BODIPY 581/591 fluorescent probe.TCGA database was used to analyze differential YAP gene expression between liver cancer and normal tissues.The effects of NaB and sorafenib on YAP and p-YAP expressions in HepG2 cells were invesitigated using Western blotting.Results NaB(2 mmol/L)significantly reduced the IC50 of sorafenib in HepG2 cells,and combination index analysis confirmed the synergy between sorafenib and NaB.The ferroptosis inhibitor Fer-1 and the YAP activator(XMU)obviously reversed the growth-inhibitory effects of the combined treatment with NaB and sorafenib in HepG2 cells.The combined treatment with NaB and sorafenib,as compared with the two agents used alone,significantly inhibited colony formation of HepG2 cells,further enhanced cellular shrinkage and dispersion,and decreased intracellular GSH and lipid ROS levels,and these effects were reversed by Fer-1 and XMU.TCGA analysis revealed a higher YAP mRNA expression in liver cancer tissues than in normal liver tissues.NaB combined with sorafenib produced significantly stronger effects than the individual agents for downregulating YAP protein expression and upregulating YAP phosphorylation level in HepG2 cells.Conclusion NaB combined with sorafenib synergistically inhibit hepatocellular carcinoma cell proliferation possibly by inducing ferroptosis via inhibiting YAP expression.

Humans ; Diabetic Nephropathies/pathology* ; Kidney/pathology* ; Biopsy ; Diabetes Mellitus, Type 2/pathology* ; Retrospective Studies ; Glomerular Filtration Rate ; Disease Progression

The interventional therapy of vascular stent implantation is a popular treatment method for cardiovascular stenosis and blockage. However, traditional stent manufacturing methods such as laser cutting are complex and cannot easily manufacture complex structures such as bifurcated stents, while three-dimensional (3D) printing technology provides a new method for manufacturing stents with complex structure and personalized designs. In this paper, a cardiovascular stent was designed, and printed using selective laser melting technology and 316L stainless steel powder of 0-10 µm size. Electrolytic polishing was performed to improve the surface quality of the printed vascular stent, and the expansion behavior of the polished stent was assessed by balloon inflation. The results showed that the newly designed cardiovascular stent could be manufactured by 3D printing technology. Electrolytic polishing removed the attached powder and reduced the surface roughness Ra from 1.36 µm to 0.82 µm. The axial shortening rate of the polished bracket was 4.23% when the outside diameter was expanded from 2.42 mm to 3.63 mm under the pressure of the balloon, and the radial rebound rate was 2.48% after unloading. The radial force of polished stent was 8.32 N. The 3D printed vascular stent can remove the surface powder through electrolytic polishing to improve the surface quality, and show good dilatation performance and radial support performance, which provides a reference for the practical application of 3D printed vascular stent.
Humans ; Stainless Steel ; Powders ; Cardiovascular System ; Constriction, Pathologic

Intensive cancer treatment with drug combination is widely exploited in the clinic but suffers from inconsistent pharmacokinetics among different therapeutic agents.To overcome it,the emerging nanomedicine offers an unparalleled opportunity for encapsulating multiple drugs in a nano-carrier.Herein,a two-step super-assembled strategy was performed to unify the pharmacokinetics of a pep-tide and a small molecular compound.In this proof-of-concept study,the bioinformatics analysis firstly revealed the potential synergies towards hepatoma therapy for the associative inhibition of exportin 1(XPO1)and ataxia telangiectasia mutated-Rad3-related(ATR),and then a super-assembled nano-pill(gold nano drug carrier loaded AZD6738 and 97-110 amino acids of apoptin(AP)(AA@G))was con-structed through camouflaging AZD6738(ATR small-molecule inhibitor)-binding human serum albumin onto the AP-Au supramolecular nanoparticle.As expected,both in vitro and in vivo experiment results verified that the AA@G possessed extraordinary biocompatibility and enhanced therapeutic effect through inducing cell cycle arrest,promoting DNA damage and inhibiting DNA repair of hepatoma cell.This work not only provides a co-delivery strategy for intensive liver cancer treatment with the clinical translational potential,but develops a common approach to unify the pharmacokinetics of peptide and small-molecular compounds,thereby extending the scope of drugs for developing the advanced com-bination therapy.

Objective:To explore the effects of intense pulsed light and carbon dioxide fractional laser sequential treatment of early hypertrophic scar after deep burn.Methods:A retrospective cohort study was used. The patients with early hypertrophic scar after deep burn who were admitted to the First People’s Hospital of Zhengzhou from May 2019 to January 2021 and met the inclusion criteria were selected as the study subjects. All patients began to receive sequential laser treatment 4-8 weeks after complete healing of wounds. The treatment method was selected according to the Vancouver scar scale (VSS) score before each treatment. If the blood vessel distribution ≥ 2 points and the thickness<2 points, they were treated with intense pulsed light. If the blood vessel distribution ≥2 points and the thickness ≥ 2 points, they were treated with intense pulsed light combined with carbon dioxide laser. If the blood vessel distribution <2 points and the thickness ≥ 2 points, they were treated with carbon dioxide laser. If the blood vessel distribution < 2 points and the thickness < 2 points, the treatment was ended. Intense pulsed light therapy was performed once a month, and carbon dioxide laser therapy was performed once every 3 months. Before and after treatment, patients were evaluated with VSS, observer scar assessment scale (OSAS) and patient scar assessment scale (PSAS), while higher scores indicated more severe scars. The number of intense pulsed light and carbon dioxide laser treatment during the treatment period, the time of scar formation and the occurrence of complications at the end of the treatment were recorded. SPSS 22.0 software was used for statistical analysis. Measurement data were expressed as Mean±SD, and paired sample t-test was used to compare patients before and after treatment. Results:A total of 28 patients were included, including 16 males and 12 females, aged 12-54 years. After the sequential treatment, the VSS scores of color, thickness, vascular distribution, softness and total score were significantly lower than those before the treatment ( t=15.00, 11.90, 15.59, 9.46, 39.24, P<0.001); OSAS scores of vascular distribution, color, thickness, roughness, softness, surface area, overall evaluation and total score were significantly lower than those before treatment ( t=14.89, 10.82, 9.54, 7.23, 16.97, 8.60, 16.42, 25.08, P<0.001); PSAS scores of pain, itching, color, hardness, thickness, irregularity, overall evaluation and total score were significantly lower than those before treatment ( t=26.40, 24.53, 16.54, 12.18, 12.25, 21.04, 22.00, 29.38, P<0.001). During the treatment, the patients were treated with intense pulsed light for (4.00±1.22) times (2-6 times), carbon dioxide laser for (2.54±1.00) times (0-5 times). At the end of the treatment, the scar formation time was (13.82±2.98) months (8-20 months). Complications occurred in 5 cases during treatment and follow-up, including 4 cases of skin blisters and 1 case of infection. No immediate skin lesions, pigmentation, depigmentation, scar aggravation and other adverse reactions occurred. Conclusion:The combination of sequential therapy of intense pulsed light and carbon dioxide laser can significantly improve the appearance and texture of early hypertrophic scar after deep burn, which has good safety.

Objective:To explore the effects of intense pulsed light and carbon dioxide fractional laser sequential treatment of early hypertrophic scar after deep burn.Methods:A retrospective cohort study was used. The patients with early hypertrophic scar after deep burn who were admitted to the First People’s Hospital of Zhengzhou from May 2019 to January 2021 and met the inclusion criteria were selected as the study subjects. All patients began to receive sequential laser treatment 4-8 weeks after complete healing of wounds. The treatment method was selected according to the Vancouver scar scale (VSS) score before each treatment. If the blood vessel distribution ≥ 2 points and the thickness<2 points, they were treated with intense pulsed light. If the blood vessel distribution ≥2 points and the thickness ≥ 2 points, they were treated with intense pulsed light combined with carbon dioxide laser. If the blood vessel distribution <2 points and the thickness ≥ 2 points, they were treated with carbon dioxide laser. If the blood vessel distribution < 2 points and the thickness < 2 points, the treatment was ended. Intense pulsed light therapy was performed once a month, and carbon dioxide laser therapy was performed once every 3 months. Before and after treatment, patients were evaluated with VSS, observer scar assessment scale (OSAS) and patient scar assessment scale (PSAS), while higher scores indicated more severe scars. The number of intense pulsed light and carbon dioxide laser treatment during the treatment period, the time of scar formation and the occurrence of complications at the end of the treatment were recorded. SPSS 22.0 software was used for statistical analysis. Measurement data were expressed as Mean±SD, and paired sample t-test was used to compare patients before and after treatment. Results:A total of 28 patients were included, including 16 males and 12 females, aged 12-54 years. After the sequential treatment, the VSS scores of color, thickness, vascular distribution, softness and total score were significantly lower than those before the treatment ( t=15.00, 11.90, 15.59, 9.46, 39.24, P<0.001); OSAS scores of vascular distribution, color, thickness, roughness, softness, surface area, overall evaluation and total score were significantly lower than those before treatment ( t=14.89, 10.82, 9.54, 7.23, 16.97, 8.60, 16.42, 25.08, P<0.001); PSAS scores of pain, itching, color, hardness, thickness, irregularity, overall evaluation and total score were significantly lower than those before treatment ( t=26.40, 24.53, 16.54, 12.18, 12.25, 21.04, 22.00, 29.38, P<0.001). During the treatment, the patients were treated with intense pulsed light for (4.00±1.22) times (2-6 times), carbon dioxide laser for (2.54±1.00) times (0-5 times). At the end of the treatment, the scar formation time was (13.82±2.98) months (8-20 months). Complications occurred in 5 cases during treatment and follow-up, including 4 cases of skin blisters and 1 case of infection. No immediate skin lesions, pigmentation, depigmentation, scar aggravation and other adverse reactions occurred. Conclusion:The combination of sequential therapy of intense pulsed light and carbon dioxide laser can significantly improve the appearance and texture of early hypertrophic scar after deep burn, which has good safety.

Objective:To investigate the clinicopathological characteristics of renal leukocyte chemotactic factor 2 amyloidosis (ALECT2).Methods:The patients with renal ALECT2 diagnosed by renal biopsy in Peking University First Hospital, Shanxi Medical University Second Hospital and Shanxi Bethune Hospital from January 2001 to October 2021 were retrospectively enrolled. According to whether the patients had concurrent glomerular diseases, they were classified into two groups: isolated ALECT2 group and ALECT2 with concurrent renal diseases group. Clinicopathological data of the two groups were compared. Light microscopy, immunofluorescence and immunoelectron microscopy were applied to investigate pathological characteristics of renal tissues. Mass spectrometry was used to analyze the composition of renal amyloid deposits. Gene sequencing was employed to detect the leukocyte chemotactic factor 2 ( LECT2) gene sequence in peripheral blood of the patients. Results:Sixteen patients with ALECT2 were enrolled in this study and nine of them had concurrent renal diseases. The age of 16 patients was (65.00±8.45) years old. The sex ratio of males to females was 7 to 9. Most of patients were Han ethnicity (15/16). Eight patients came from Shanxi province. Fifteen patients presented with varying degree of proteinuria [2.16(1.07, 4.72) g/24 h]; 5 patients had nephrotic syndrome; 11 patients had renal insufficiency; 12 patients had microscopic hematuria. Part of patients also had hypertension (12/16) and diabetics (6/16). Compared with isolated ALECT2, the ALECT2 group with concurrent renal diseases had a higher proportion of nephrotic syndrome (5/9 vs 0/7, P=0.034). Renal biopsy results showed that all patients (16/16) had amyloid deposits in the interstitium of renal cortex with varying degree of inflammatory cell infiltration and fibrosis, and glomeruli (12/16) and arterioles (14/16) were involved by amyloid deposits. The amyloid deposits were strongly congophilic and immunohistochemistry for LECT2 was positive. By semi-quantitative analysis, the proportions of glomerular and overall amyloid loads in ALECT2 with concurrent renal diseases group were lower than those in isolated ALECT2 group (both P<0.05). Electron microscopy revealed randomly oriented and non-branching fibrils with a diameter of 8-12 nm. The LECT2 peptides were detected by mass spectrometry in renal amyloid deposits of 8 patients, and homozygous G allele of LECT2 was found in 7 patients by gene sequencing. Complete follow-up data of 13 patients showed that 2 patients died, 1 patient developed end-stage renal disease at the time of renal biopsy, and most of the rest patients had stable renal function (8/10). Conclusions:Patients with renal ALECT2 mainly present with proteinuria, along with a high incidence of renal insufficiency, microscopic hematuria, and concurrent renal diseases. The pathologic feature is the preferential deposition of amyloid in renal cortical interstitium.

Background: Symptoms of irritable bowel syndrome with diarrhea (IBS-D) are known to be influenced by circadian oscillation; however, the pathophysiological mechanism is still unclear. Aims: To investigate the role and underlying mechanism of colon circadian clock gene Bmal1 involved in the occurrence of symptoms in IBS-D patients. Methods: Forty-six patients with IBS-D and 34 normal controls from Army Medical Center of PLA during September 2018 to February 2021 were recruited in this study. IBS-severity scoring system (IBS-SSS) was used to evaluate the severity of IBS-D symptoms. A colonoscopy was performed to obtain biopsy specimens from rectosigmoid colon. The concentration of 5-hydroxytryptamine (5-HT), and expressions of Bmal1 and chromogranin A (CgA), a biomarker of enterochromaffin cells (EC cells), in colonic mucosa were detected by ELISA and double-labeled immunofluorescence, respectively. Results: Both the 5-HT concentration and number of EC cells in colonic mucosa of IBS-D patients were significantly higher than those of the normal controls (all P< 0.05). Bmal1 was mainly expressed in intestinal epithelial cells and was highly expressed in EC cells. Co-expression of Bmal1 and CgA was observed. Compared with the normal control group biopsied at the same time point, expression of Bmal1 was significantly higher in specimens taken at 9 a.m., and expression of Bmal1 was significantly lower in specimens taken at 17 p.m. in IBS-D patients (all P< 0.05). Spearman correlation coefficient analysis showed that Bmal1 expression at 9 a.m. was positively correlated with the total score of IBS-SSS and subscore of abdominal pain and discomfort (r