Mechanism of BMAL1 attenuating H2O2-induced cardiomyocyte injury / 天津医药

Objective To investigate the effect of BMAL1 on H2O2-induced cardiomyocyte injury through NRF2-regulated ROS/NLRP3 inflammasome pathway.Methods H9c2 cells and H9c2 cells with stable over-expressed BMAL1 were cultured and divided into the control group,the H2O2 group,the BMAL1-OE group,the BMAL1-OE+H2O2 group,the BMAL1-OE+ML385 group and the BMAL1-OE+ML385+H2O2 group.All groups were pre-intervened with corresponding inhibitors,and then treated with 0.2 mmol/L H2O2,except for the control group and the BMAL1-OE group.After the intervention,CCK-8 assay was used to measure cell viability,fluorescent probe DCFH-DA was used to measure ROS generation and Western blot assay was used to detect BMAL1,NRF2 and NLRP3 protein expressions.ELISA was used to determine IL-1β release.Results Compared with the control group,the cell viability was decreased,ROS generation was increased,BMAL1 and NRF2 protein expressions were decreased,NLRP3 expression and IL-1β release were increased in the H2O2 group(P＜0.05).Compared with the H2O2 group,the cell viability was increased,ROS generation was decreased,BMAL1-OE and NRF2 protein expressions were increased,NLRP3 expression and IL-1β release were decreased in the BMAL1-OE+H2O2 group(P＜0.05).Compared with the BMAL1-OE+H2O2 group,the cell viability was decreased,ROS generation was increased,NLRP3 expression and IL-1β release were increased in the BMAL1-OE+ML385+H2O2 group(P＜0.05).Conclusion BMAL1 attenuates H2O2-induced H9c2 cardiomyocyte injury,and its mechanism may be related to the regulation of ROS/NLRP3 inflammasome pathway through NRF2.

Screening and analysis of differentially expressed genes for calcium homeostasis in ameloblasts with high fluoride intervention / 中国组织工程研究

BACKGROUND:Fluorosis is a disorder of enamel development caused by long-term intake of large amounts of fluoride during enamel development. OBJECTIVE:To further explore the molecular mechanism of dental fluorosis formation by screening the differentially expressed genes associated with calcium homeostasis in ameloblasts by transcriptome sequencing technology. METHODS:LS8 cells were treated with 0,0.4,0.8,1.6,3.2 and 6.4 mmol/L sodium fluoride(NaF)for 24,48 and 72 hours to observe the effects of different concentrations of NaF on the morphology,cell activity and intracellular Ca2+ concentration of LS8 cells.The differentially expressed genes were screened by transcriptome sequencing and validated. RESULTS AND CONCLUSION:After 24 hours of treatment,the cells treated with 0,0.4,and 0.8 mmol/L NaF were in good growth condition,with increased cell number and clear cell outline.When the NaF concentration was≥1.6 mmol/L,the cells were gradually shrunken and became smaller and the number of cells decreased with the increase of NaF concentration.After 48 and 72 hours of treatment,the number of cells increased in the 0,0.4 mmol/L NaF groups,while gradually decreased in the 0.8,1.6,3.2 mmol/L NaF groups,with rounded and smaller cell morphology.The cells in the 6.4 mmol/L NaF group were shrunken,rounded and suspended in the medium,with almost no adherent cells.When treated with the same concentration of NaF,LS8 cells were in optimal growth after 24 hours of treatment.Results from cell counting kit-8 assay showed that when treated with the same concentration of NaF,the cell activity decreased with the increase of treatment time;when the treatment time was the same,the cell activity decreased with the increase of NaF concentration.After 24 hours of treatment,the intracellular Ca2+ concentration increased with the increase of NaF concentration.Transcriptome sequencing analysis identified genes involved in the regulation of cellular calcium homeostasis:Hsp90b1,Canx,Calr,and Hspa5 that were significantly upregulated(P＜0.05)and Cacna1a that was significantly downregulated(P＜0.05).To conclude,the inhibitory effect of NaF on LS8 cell proliferation may be related to the abnormal increase in intracellular Ca2+ concentration,and the mechanism may be caused by the upregulation of the expression of protein processing and synthesis pathways Hsp90b1,Canx,Calr,and Hspa5 and the downregulation of the expression of calcium signaling pathway Cacna1a.

Screening and analysis of ferroptosis related genes in peripheral blood mononuclear cells of patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective:To analyze the differentially expressed genes in PBMCs of patients with systemic lupus erythematosus (SLE) by bioinformatics methods screening and analyzing the key genes related to ferroptosis, and explore the possible mechanism of ferroptosis involved in the pathogenesis of SLE at the transcription level.Methods:The data sets and samples of healthy people (HC) and SLE patients who met the screening criteria were retrieved from the Gene Expression Omnibus (GEO), a sub-database of the National Center for Biotechnology Information (NCBI). The differentially expressed genes, GO enrichment analysis and KEGG pathway enrichment analysis were analyzed by GEO2R, R language and related software packages. The protein interaction network (PPI) of differential genes was analyzed by STRING, Cytoscape and other tools to explore the key genes and pathways. In addition, real-time quantitative reverse transcription PCR (RT-qPCR) was used to verify the expression of key genes. Mann-Whitney U test was used to compare the expression of key genes in PBMCs between the two groups. Spearman rank correlation analysis was used to explore the relationship between SLE disease activity and the level of key genes. Results:Six data sets were included in this study. A total of 166 genes related to ferroptosis were differentially expressed between SLE and HC groups. The differential genes were specifically expressed in alveolar macrophages, neutrophils, CD49 + cells and CD31 + cells. GO enrichment analysis and KEGG pathway enrichment analysis showed that the differentially expressed genes were mainly involved in multiple signaling pathways closely related to SLE, such as oxidative stress response, infection and TNF signaling pathway. Hub genes screened by different algorithms all suggested RELA as a key gene, and RT-qPCR confirmed that compared with the RELA gene expression level in the HC group [0.75(0.37,1.13)], the expression level in SLE group [2.02 (1.19,4.06)] was increased, the difference was statistically significant ( Z=-3.08, P=0.002), and was positively correlated with the corresponding SLEDAI score of SLE samples ( r=0.52, P=0.019). Conclusion:The ferroptosis of many immune cells, including alveolar macrophages and CD49 + NK cells, is involved in the pathogenesis of SLE. RELA may be involved in the ferroptosis of PBMCs in SLE through the NF-κB pathway.

Feasibility of treatment planning for 4D-CT high ventilation functional lung avoided radiotherapy in thoracic cancer / 中华放射医学与防护杂志

Objective:To establish a radiotherapy treatment planning process of high ventilation functional lung avoided (HVFLA) for thoracic tumors based on 4D-CT lung ventilation functional images and determine the treatment planning strategy of HVFLA radiotherapy, and so as to provide support for the clinical trials of HVFLA radiotherapy in thoracic cancer patients.Methods:A deep learning-based 4D-CT lung ventilation functional imaging model was established and integrated into the radiotherapy treatment planning process. Furthermore, ten thoracic cancer patients with 4D-CT simulation positioning were retrospectively enrolled in this study. The established model was used to obtain the 4D-CT lung ventilation functional imaging for each patient. According to the relative value of lung ventilation, the lung ventilation areas are equally segmented into high, medium and low lung ventilation and then imported them into Pinnacle 3 treatment planning system. According to the prescription dose of target and dose constraints of organ at risks (OARs), the clinical and HVFLA treatment plans were designed for each patient using volumetric modulated radiotherapy technique, and each plan should meet the clinical requirements and adding dose constraints of high ventilation functional lung for HVFLA plan. The dosimetric indexes of the target, OARs (lungs, heart and cord) and high functional lung (HFL) were used to evaluated the plan quality. The dosimetric indexes included D2, D98 and mean dose of target, V5, V10, V20, V30 and mean dose of lungs and HFL, V30, V40 and mean dose of heart, and D1 cm 3 of cord. Paired samples t-test was used for statistical analysis of the two groups of plans. Results:The target and OARs of the clinical plan and HVFLA plan meet the clinical requirements. The HVFLA plan resulted in a statistically significant reduction in the mean dose, V5, V10, V20, and V30 of the high functional lung by 1.2 Gy, 5.9%, 4.2%, 2.6%, and 2.3%, respectively ( t=-8.07, 4.02, -6.02, -7.06, -6.77, P<0.05). There was no statistical difference in the dosimetric indexes of lungs, heart and cord. Conclusions:We established the treatment planning process of HVFLA radiotherapy based on 4D-CT lung ventilation functional images. The HVFLA plan can effectively reduce the dose of HFL, while the doses of lungs, heart and cord had no significant difference compared with the clinical plan. The strategy of HVFLA radiotherapy planning is feasible to provide support for the implementation of HVFLA radiotherapy in thoracic cancer patients.

Feasibility of acceptance of multiple accelerators using Elekta AGL standard procedures / 中华放射肿瘤学杂志

Objective:To verify the feasibility of using Elekta accelerated go live (AGL) standard process for the acceptance of multiple accelerators.Methods:The beams of three accelerators were adjusted by PTW Beamscan three-dimensional water tank to reach the AGL standard. Dose verification was performed for three accelerators that met AGL standards. A simple field test example from Cancer Hospital Chinese Academy of Medical Sciences was used to compare the MapCheck 3 surface dose measurement results with the surface dose calculated by the same accelerator model. Images of 10 patients including head and neck, esophagus, breast, lung and rectum were randomly selected. volumetric-modulated arc therapy (VMAT) and intensity modulated radiation therapy (IMRT) treatment techniques were used for planning design, and the measured dose of ArcCheck was compared with the planned dose calculated by the same accelerator model. One-way ANOVA was used to statistically analyze the passing rates of two-dimensional and three-dimensional dose verification.Results:The 6 MV X-ray percentage depth dose at 10 cm underwater (PDD 10) of three accelerators was 67.45%, 67.36%, 67.47%, and the maximum deviation between the three accelerators was 0.11%. The 6 MV flattenting filter free (FFF) mode X-ray PDD 10 was 67.33%, 67.20%, 67.20%, and the maximum deviation between the three accelerators was 0.13%. All required discrete point doses on each energy 30 cm×30 cm Profile spindle of the three accelerator X-rays deviated less than ±1% from the standard data. Absolute γ analysis was performed on the results of MapCheck 3 two-dimensional dose matrix validation. Under the 10% threshold of 2 mm/3% standard, the average passing rate of the test cases in Cancer Hospital Chinese Academy of Medical Sciences was above 99%, and the difference was not statistically significant ( P>0.05). Absolute γ analysis was performed on the ArcCheck verification results. Under the 10% threshold, the pass rate of 2 mm/3% was all above 95%, the maximum average passing rate of the three accelerators with different energy and different treatment techniques was 0.28% (6 MV, VMAT), 0.19%(6 MV FFF, VMAT), 0.56% (6 MV, IMRT) and 0.05% (6 MV FFF, IMRT), and the difference was not statistically significant ( P>0.05). Conclusion:Compared with traditional accelerator acceptance process, the acceptance time of each accelerator is shortened by 4-6 weeks by using the AGL standard process, and the radiotherapy plan of patients can be interchangeably executed among different accelerators.

Establishment and evaluation of a RAA-CRISPR-Cas13a method for detecting KPC carbapenemase genes / 中华检验医学杂志

Objective:To establish a rapid and accurate method for the detection of Klebsiella pneumoniae carbapenemase (KPC) carbapenemase gene based on recombinase aided amplification (RAA)-CRISPR-Cas13a (CRISPR-Cas13a) technology. Methods:Twenty-five clinical isolates of carbapenem-resistant Klebsiella pneumoniae (CRKP) and five carbapenem-sensitive Klebsiella pneumoniae (CSKP) strains preserved in 2020-2021 in Beijing Chuiyangliu Hospital were randomly collected, and the total DNA samples of the strains was extracted. RAA primers specific for KPC DNA and CRISPR RNA (crRNA) were designed to establish a rapid and accurate method for the detection of KPC carbapenemase gene based on RAA-CRISPR-Cas13a technology. The method was evaluated by plasmids and clinical sample strains, and the detection was also performed by Quantitative real-time PCR (qPCR) method to compare the detection rate and consistency of the two methods. Results:The RAA-CRISPR-Cas13a method can detect KPC plasmids and samples with a sensitivity of 1 copy/μl, which is higher than that of qPCR (10 1 copies/μl). Among the 30 clinical strains (including 25 CRKP strains and 5 CSKP strains), 23 strains were detected to carry KPC gene by both RAA-CRISPR-Cas13a method and qPCR method, and 7 strains were not detected with KPC gene. The detection rate of KPC gene in the 25 CRKP strains was 92% (23/25). The positive coincidence rate of the two methods was 100% (23/23). Conclusions:This study combined RAA amplification technology with CRISPR-Cas13a technology to establish a rapid and accurate method for detecting KPC carbapenemase gene. The method is useful for accurate screening of KPC carbapenemase-producing strains. It has a wide application prospect in drug resistance monitoring and infection control.

Efficacy comparison between olaparib and platinum-containing regimen for treatment of platinum-sensitive relapsed ovarian cancer patients with BRCA mutation / 肿瘤研究与临床

Objective:To explore the differences in tumor-specific growth factors, cellular immune function and efficacy of olaparib and platinum-containing regimen for treatment of platinum-sensitive relapsed ovarian cancer patients with BRCA mutation.Methods:A retrospective cohort study was conducted. A total of 100 platinum-sensitive relapsed BRCA-mutant ovarian cancer patients in Baoding Second Central Hospital from September 2017 to March 2020 were retrospectively selected. The clinical data of the patients were analyzed, and they were divided into the olaparib group (treated with olaparib tablets) and the platinum-containing regimen group (treated with paclitaxel and platinum drugs for 6 cycles, followed by olaparib tablets maintenance therapy), with 50 patients in each group. The clinical efficacy, tumor specific growth factor [carbohydrate antigen (CA) 125, CA199, human epididymal protein 4 (HE4)] levels, cellular immune function-related indicators [T-cell subsets (proportions of CD3 + cells and CD4 + cells), CD4 + cells/CD8 + cells ratio (CD4 +/CD8 +)], and quality of life scores before treatment and after 2, 4 and 6 cycles of treatment of the two groups were compared, as well as the safety of the two groups. The data of three years of follow-up were obtained, Kaplan-Meier method was used to analyze the progression-free survival (PFS) of patients in the two groups, and log-rank test was used for comparison between groups. Results:The age of patients in the olaparib and platinum-containing regimen groups was (53±7) years old and (56±7) years old, respectively. The differences in compositions of patients with different age, body mass index, Eastern Cooperative Oncology Group (ECOG) performance status score, primary tumor location, lesion size, pathological stage, pathological type, germline BRCA mutation, and previous chemotherapy response between the two groups were not statistically significant (all P > 0.05). The objective response rate (ORR) [58.0% (29/50) vs. 38.0% (19/50)] and disease control rate (DCR) [80.0% (40/50) vs. 56.0% (28/50)] of the olaparib group after treatment were higher than those of the platinum-containing regimen group, and the differences were statistically significant (both P < 0.05). Serum CA125, CA199 and HE4 levels were gradually decreased in both groups before treatment and after 2, 4 and 6 cycles of treatment (all P < 0.05); serum CA125, CA199 and HE4 levels in the olaparib group after 2, 4 and 6 cycles of treatment were lower than those in the platinum-containing regimen group, and the differences were statistically significant (all P < 0.05). The CD3 + cells ratio, CD4 + cell ratio and CD4 +/CD8 + in the olaparib group gradually increased before treatment and after 2, 4 and 6 cycles of treatment (all P < 0.05), while those in the platinum-containing regimen group all gradually decreased (all P < 0.05); the CD3 + cells ratio, CD4 + cells ratio and CD4 +/CD8 + in the olaparib group were higher than those in the platinum-containing regimen group after 2, 4 and 6 cycles of treatment, and the differences were statistically significant (all P < 0.05). The quality of life scores of both groups increased before treatment and after 2, 4 and 6 cycles of treatment (all P < 0.05), and the quality of life scores of the olaparib group were higher than those of the platinum-containing regimen group after 2, 4 and 6 cycles of treatment, and the differences were statistically significant (all P < 0.05). The incidence of nausea, fatigue and malaise, vomiting, anemia, and diarrhea at all levels in the olaparib group was lower than those in the platinum-containing regimen group (all P < 0.05). By follow-up for 3 years, there was no statistically significant difference in PFS between the olaparib group and the platinum-containing regimen group ( P > 0.05). Conclusions:The efficacy of olaparib treatment in platinum-sensitive relapsed ovarian cancer patients with BRCA mutation is superior to platinum-containing regimen, and it can increase the level of T cells, inhibit the expression of tumor-specific growth factors, improve the quality of life, and have a positive effect on improving the safety of treatment.

Novel antibacterial drug target against Gram-negative bacteria: lipopolysaccharide transport protein LptDE and its inhibitors / 药学学报

The outer membrane composed predominantly of lipopolysaccharide (LPS) is an essential biological barrier for most Gram-negative (G-) bacteria. Lipopolysaccharide transport protein (Lpt) complex LptDE is responsible for the critical final stage of LPS transport and outer membrane assembly. The structure and function of LptDE are highly conserved in most G- bacteria but absent in mammalian cells, and thus LptDE complex is regarded as an attractive antibacterial target. In recent 10 years, the deciphering of the three-dimensional structure of LptDE protein facilities the drug discovery based on such "non-enzyme" proteins. Murepavadin, a peptidomimetic compound, was reported to be the first compound able to target LptD, enlightening a new class of antibacterial molecules with novel mechanisms of action. This article is devoted to summarize the molecular characteristics, structure-function of LptDE protein complex and review the development of murepavadin and related peptidomimetic compounds, in order to provide references for relevant researches.

Recent findings with regard to roles of GGA2 in vesicle transport and related diseases / 中国病理生理杂志

Cellular dysfunction caused by vesicle transport is associated with a variety of diseases.The trans-Golgi network(TGN)to endosome transport is an important pathway of vesicle transport,and its defects leading to protein balance disorders has been linked to many diseases such as cancer,neurodegenerative diseases and diabetes mellitus.Gol-gi-associated gamma-adaptin ear-containing ADP-ribosylation factor-binding protein 2(GGA2)is a crucial protein in-volved in TGN-endosomal transport.It plays a significant role in the regulation of several diseases,including cancer,Al-zheimer disease,type 2 diabetes mellitus and cerebral ischemia,by mediating protein transport with important biological significance.This article provides an overview of the molecular structure of GGA2,its role in regulating clathrin-mediated protein transport between TGN and endosomes,and its potential implications for a variety of diseases.

Elevated Levels of Naturally-Occurring Autoantibodies Against the Extracellular Domain of p75NTR Aggravate the Pathology of Alzheimer's Disease / 神经科学通报·英文版

The extracellular domain (p75ECD) of p75 neurotrophin receptor (p75NTR) antagonizes Aβ neurotoxicity and promotes Aβ clearance in Alzheimer's disease (AD). The impaired shedding of p75ECD is a key pathological process in AD, but its regulatory mechanism is largely unknown. This study was designed to investigate the presence and alterations of naturally-occurring autoantibodies against p75ECD (p75ECD-NAbs) in AD patients and their effects on AD pathology. We found that the cerebrospinal fluid (CSF) level of p75ECD-NAbs was increased in AD, and negatively associated with the CSF levels of p75ECD. Transgenic AD mice actively immunized with p75ECD showed a lower level of p75ECD and more severe AD pathology in the brain, as well as worse cognitive functions than the control groups, which were immunized with Re-p75ECD (the reverse sequence of p75ECD) and phosphate-buffered saline, respectively. These findings demonstrate the impact of p75ECD-NAbs on p75NTR/p75ECD imbalance, providing a novel insight into the role of autoimmunity and p75NTR in AD.

Mechanisms of cholesterol metabolism imbalance in a PA-induced non-alcoholic fatty liver disease cell model / 中国药科大学学报

@#Liver cholesterol metabolism disorder plays an important role in the development of non-alcoholic fatty liver disease (NAFLD).In order to reveal the molecular mechanism of cholesterol homeostasis imbalance induced by saturated fatty acids, HepG2 cells were stimulated with palmitic acid (PA).Lipids accumulation was analyzed by Oil Red O staining, intracellular triglyceride and cholesterol quantification.The level of genes and proteins related to cholesterol homeostasis was measured by RT-qPCR and western blotting.Additionally, intracellular bile acids and mitochondrial oxysterols were detected by LC-MS/MS.The results demonstrated that intracellular lipids such as TG and TC were significantly increased in the model with PA stimulation.Although no significant difference was detected in genes related to cholesterol synthesis and uptake, the protein expression of ABCG5 and LXRα were significantly down-regulated, indicating a decrease in cholesterol efflux.Meanwhile, the gene expression of STARD1 and CYP7B1, which are responsible for bile acid alternative synthesis, were markedly enhanced, along with a significant increase of cholesterol and 27-OHC in mitochondria and CDCA in cells.These results suggested that PA overload may disrupt cholesterol homeostasis by inhibiting cholesterol efflux and promoting bile acids synthesis.

N6-Methyladenosine Methyltransferase METTL3 Alleviates Diabetes-Induced Testicular Damage through Modulating TUG1/Clusterin Axis

Background@#The present study investigated the regulatory effects of N6-methyladenosine (m6A) methyltransferase like-3 (METTL3) in diabetes-induced testicular damage. @*Methods@#In vivo diabetic mice and high glucose (HG) treated GC-1 spg cells were established. The mRNA and protein expressions were determined by real-time quantitative polymerase chain reaction, Western blot, immunofluorescence and immunohistochemistry staining. Levels of testosterone, blood glucose, cell viability, and apoptosis were detected by enzyme-linked immunosorbent assay, MTT, and flow cytometry, respectively. Molecular interactions were verified by RNA immunoprecipitation and RNA pull-down assay. Histopathological staining was performed to evaluate testicular injury. @*Results@#METTL3 and long non-coding RNA taurine up-regulated 1 (lncRNA TUG1) were downregulated in testicular tissues of diabetic mice and HG-treated GC-1 spg cells. METTL3 overexpression could reduce the blood glucose level, oxidative stress and testicular damage but enhance testosterone secretion in diabetic mouse model and HG-stimulated GC-1 spg cells. Mechanically, METTL3-mediated m6A methylation enhanced the stability of TUG1, then stabilizing the clusterin mRNA via recruiting serine and arginine rich splicing factor 1. Moreover, inhibition of TUG1/clusterin signaling markedly reversed the protective impacts of METTL3 overexpression on HG-stimulated GC-1 spg cells. @*Conclusion@#This study demonstrated that METTL3 ameliorated diabetes-induced testicular damage by upregulating the TUG1/clusterin signaling. These data further elucidate the potential regulatory mechanisms of m6A modification on diabetes-induced testicular injury.

Study the change of serum osteosclerotin and Dickkopf-3 expression in elderly patients with intertrochanteric fracture / 中国医师进修杂志

Objective:To investigate the changes of perioperative serum osteosclerosis protein (SOST) and Dickkopf-3 (Dkk-3) in elderly patients with femoral intertrochanteric fracture.Methods:Thirty elderly patients who underwent reduction and fixation of femoral intertrochanteric fracture in Baoding Second Hospital from May 2017 to December 2017 were prospectively selected as the observation group; 30 healthy subjects in the same period were selected as the healthy control group. Enzyme linked immunosorbent assay (ELISA) was used to detect the expression of serum SOST and Dkk-3 at 1 d before operation and at 1, 3, 5 d after operation and compared with the same period of healthy physical examination(normal control group). Spearman rank correlation analysis was used to analyze the correlation between SOST and Dkk-3 and disease activity score (ASDAS) and spinal imaging evaluation score (mSASSS).Results:There was a positive correlation between Dkk-3 level and ASDAS score in the observation group ( r = 0.331, P = 0.012); the level of SOST was negatively correlated with the scores of ASDAS ( r = - 0.162, P = 0.017). The levels of serum SOST and Dkk-3 in the observation group were lower than those in the healthy control group: 1.29(1.00, 2.40) μg/L vs. 1.96(1.63, 2.65) μg/L, (6.11 ± 1.15) μg/L vs. (9.81 ± 1.76) μg/L, P<0.05. The levels of serum SOST and Dkk-3 in the observation group increased first and then decreased on the 1st, 3rd and 5th day after operation. The level of serum Dkk-3 increased to the highest level on the 3rd day after operation, and then decreased gradually, but it was still slightly higher than that before operation. The level of serum SOST in the observation group increased to the highest level 1st day after operation, and decreased at 3rd and 5th day after operation. The perioperative serum levels of SOST and Dkk-3 in the observation group were positively correlated, the correlation coefficient was the largest at 1 day after operation ( r = 0.571) and the lowest before operation ( r = 0.119). Conclusions:The perioperative serum levels of SOST and Dkk-3 in elderly patients with femoral intertrochanteric fracture increased first and then decreased. The change of serum SOST level is more sensitive and can be used as a sensitive index to reflect the change of osteogenic ability.

Application of a novel modified Blumgart anastomosis in laparoscopic pancreaticoduodenectomy / 中国医师进修杂志

Objective:To investigate the efficacy and safety of a novel modified Blumgart pancreaticojejunostomy in laparoscopic pancreaticoduodenectomy (LPD).Methods:Between May 2021 and January 2022, 13 successive cases from Lihuili Hospital Affiliated to Ningbo University who underwent LPD were enrolled in this retrospective study. The study retrospectively analyzed the demographic characteristics, perioperative outcomes, and pathological results of these cases.Results:Twenty patients underwent LPD success-fully and one required conversion to open surgery. The operative time was (308.6 ± 61.7) min. The duration for PJ was (26.7 ± 4.3) min. The estimated blood loss was (188.1 ± 94.2) ml. The postoperative hospital stay was (14.2 ± 3.5) d. There was one case of biochemical leakage and no case of grade B or grade C pancreatic fistula.Conclusions:The new method is safe, simple and feasible. The novel method could reduce the incidence of pancreatic fistula and other complications after LPD.

Establishment of prediction model for severe acute pancreatitis complicated with abdominal hypertension / 中华胰腺病杂志

Objective:To construct the prediction model of SAP complicated with intra-abdominal hypertension (IAH), and evaluate the prediction efficiency of the model.Methods:The clinical data of 322 SAP patients admitted to the emergency department of Cangzhou Hospital of Integrated Chinese and Western Medicine in Hebei Province from January 2017 to December 2021 were retrospectively analyzed. They were divided into IAH group ( n=153) and control group ( n=169) according to whether they had IAH complications or not. The clinical characteristics and laboratory test results of the two groups were compared. Multifactor logistic step-up regression was used to analyze the risk factors of SAP patients complicated with IAH. A nomogram model for predicting SAP complicated with IAH was established by using R software. The receiver operating characteristic curve (ROC) of the model was plotted, and the area under the curve (AUC) was calculated to evaluate its prediction efficiency. Calibration chart, Hosmer-Lemesshow test and decision curve analysis were used to evaluate the prediction accuracy and clinical application value of the model. The Bootstrap method was applied to verify the model internally. Results:In IAH group, cases with body mass index, CRP, procalcitonin (PCT), WBC, acute physiological and chronic health assessmentⅡ (APACHEⅡ) score, modified CT Severity Index score (MCTSI), incidence of complications (abdominal effusion, abdominal infection, gastrointestinal dysfunction, shock, multiple organ dysfunction syndrome), mechanical ventilation, the number of high-volume fluid reactivation (24 h≥4 L) were more than those in control group; serum albumin and serum calcium in IAH group were lower than those in control group, and the differences were statistically significant (all P value <0.05). Multivariate logistic regression analysis showed that serum albumin ( OR=0.815, 95% CI 0.710-0.937), CRP ( OR=1.005, 95% CI 1.002-1.008), MCTSI ( OR=2.043, 95% CI 1.695-2.463), complication of gastrointestinal dysfunction ( OR=4.179, 95% CI 2.170-8.049), and high-volume fluid resuscitation ( OR=4.265, 95% CI 2.269-8.015) were independent risk factors for IAH in SAP.The Nomogram prediction model was established using the five factors above as parameters, and the AUC value for predicting IAH complication was 0.886. The Hosmer-Lemesshow test showed a high consistency between the prediction results and the actual clinical observation results ( P=0.189). The results of decision curve analysis showed that the prediction probability of the model was between 10% and 85%, which could bring more benefits to patients. Conclusions:The early prediction model of SAP with concurrent IAH is successfully established, which can better predict the risk of SAP with concurrent IAH.

Progress in molecular signal pathway of dental fluorosis / 中华地方病学杂志

Dental fluorosis is a manifestation of chronic oral fluorosis caused by excessive fluoride intake in childhood. At present, the molecular mechanism of dental fluorosis is still unclear. Ameloblasts are the most sensitive cells to fluoride in tooth tissue. Fluoride affects the proliferation and secretion of ameloblasts through the role of key molecules in the molecular signal pathways, leading to the formation of dental fluorosis. This paper reviews the relationship between the molecular signal pathways [mitogen-activated protein kinase (MAPK), transforming growth factor-β (TGF-β)/Smad, Wnt, Foxo1/Runx2], stress pathways (endoplasmic reticulum stress and oxidative stress) and the occurrence of dental fluorosis in recent years, in order to deeply understand the pathogenesis of dental fluorosis at the molecular level, and provide new ideas for the prevention and treatment of dental fluorosis.

Effect of Plasmacytoid Dendritic Cell Dose in Grafts on CMV Infection after Allogeneic Hematopoietic Stem Cell Transplantation / 中国实验血液学杂志

OBJECTIVE@#To investigate the correlation between plasmacytoid dendritic cell (pDC) dose in grafts and the occurrence of cytomegalovirus (CMV) infection after allogeneic hematopoietic stem cell transplantation (allo-HSCT).@*METHODS@#The clinical data of 80 children who received allo-HSCT in Children's Hospital of Soochow University from August 20, 2020 to June 11, 2021 were retrospectively analyzed. Proportions of DC subsets and T-cell subsets in grafts were detected by flow cytometry in order to calculate infused cell dose of each cell. Weekly monitoring of CMV-DNA copies in peripheral blood for each child were performed after transplantation. The last follow-up date was December 31, 2021.@*RESULTS@#All the children gained hematopoietic reconstitution. CMV infection was observed in 51 children (63.8%±5.4%) within the first 100 days after transplantation, including 2 cases developing CMV disease. Univariate analysis indicated that infused doses of DC and pDC were significantly associated with CMV infection within 100 days after allo-HSCT (P <0.05). Multivariate analysis indicated that a high dose infusion of pDC was an independent protective factor for CMV infection within 100 days after allo-HSCT (P <0.05). By the end of follow-up, 7 children died of transplantation-related complications, including 2 deaths from CMV disease, 2 deaths from extensive chronic graft-versus-host disease, and 3 deaths from capillary leak syndrome. The overall survival rate was 91.2%.@*CONCLUSION@#The pDC in grafts may be associated with early infection of CMV after allo-HSCT, while a high infused pDC dose may serve as a protective factor for CMV infection after transplantation.

Mechanism of Zhenwu Decoction in improving renal inflammatory injury in mice with DN of spleen-kidney Yang deficiency syndrome by regulating ROCK/IKK/NF-κB pathway / 中国中药杂志

To investigate the intervention effect and mechanism of Zhenwu Decoction on diabetic nephropathy(DN) mice of spleen-kidney Yang deficiency syndrome based on the Rho-associated coiled-coil kinase(ROCK)/IκB kinase(IKK)/nuclear factor-κB(NF-κB) pathway. Ninety-five 7-week-old db/db male mice and 25 7-week-old db/m male mice were fed adaptively for one week. The DN model of spleen-kidney Yang deficiency syndrome was induced by Dahuang Decoction combined with hydrocortisone by gavage, and then the model was evaluated. After modeling, they were randomly divided into a model group, high-dose, medium-dose, and low-dose Zhenwu Decoction groups(33.8, 16.9, and 8.45 g·kg~(-1)·d~(-1)), and an irbesartan group(25 mg·kg~(-1)·d~(-1)), with at least 15 animals in each group. The intervention lasted for eight weeks. After the intervention, body weight and food intake were measured. Serum crea-tinine(Scr), blood urea nitrogen(BUN), fasting blood glucose(FBG), urinary albumin(uALb), and urine creatinine(Ucr) were determined. The uALb/Ucr ratio(ACR) and 24 h urinary protein(UTP) were calculated. Renal pathological morphology was evaluated by HE staining and Masson staining. The levels of key molecular proteins in the ROCK/IKK/NF-κB pathway were detected by Western blot. Enzyme-linked immunosorbent assay(ELISA) was used to detect interleukin-1β(IL-1β), interleukin-6(IL-6), interleukin-8(IL-8), interleukin-10(IL-10), and tumor necrosis factor-α(TNF-α). Compared with the blank group, the model group showed increased content of BUN, uALb, and SCr, increased values of 24 h UTP and ACR, decreased content of Ucr(P<0.05), enlarged glomeruli, thickened basement membrane, mesangial matrix proliferation, inflammatory cell infiltration, and collagen fiber deposition. The protein expression of ROCK1, ROCK2, IKK, NF-κB, phosphorylated IKK(p-IKK), phosphorylated NF-κB(p-NF-κB), and phosphorylated inhibitor of NF-κB(p-IκB) increased(P<0.05), while the protein expression of inhibitor of NF-κB(IκB) decreased(P<0.05). The levels of inflammatory factors IL-1β, IL-6, IL-8, and TNF-α increased(P<0.05), while the level of IL-10 decreased(P<0.05). Compared with the model group, the groups with drug treatment showed decreased levels of BUN, uALb, SCr, 24 h UTP, and ACR, increased level of Ucr(P<0.05), and improved renal pathological status to varying degrees. The high-and medium-dose Zhenwu Decoction groups and the irbesartan group showed reduced protein expression of ROCK1, ROCK2, IKK, NF-κB, p-IKK, p-NF-κB, and p-IκB in the kidneys(P<0.05), increased protein expression of IκB(P<0.05), decreased levels of serum inflammatory factors IL-1β, IL-6, IL-8, and TNF-α(P<0.05), and increased level of IL-10(P<0.05). Zhenwu Decoction can significantly improve renal function and renal pathological damage in DN mice of spleen-kidney Yang deficiency syndrome, and its specific mechanism may be related to the inhibition of inflammatory response by down-regulating the expression of key molecules in the ROCK/IKK/NF-κB pathway in the kidney.

Analysis of ARX gene variant in a child with X-linked lissencephaly with abnormal genitalia / 中华医学遗传学杂志

OBJECTIVE@#To explore the clinical characteristics and genetic basis for a child with X-linked lissencephaly with abnormal genitalia (XLAG).@*METHODS@#A child with XLAG who had presented at the Third Affiliated Hospital of Zhengzhou University in May 2021 was selected as the study subject. Peripheral blood samples of the child and his parents were collected and subjected to high-throughput sequencing. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the result was analyzed by using bioinformatic software.@*RESULTS@#The child was found to have harbored a hemizygous c.945_948del variant in exon 2 of the ARX gene, which as a frameshifting variant has resulted in a truncated protein. His mother was found to be heterozygous for the variant, whilst his father was of wild type. The variant was unreported previously.@*CONCLUSION@#The hemizygous c.945_948del variant of the ARX gene probably underlay the XLAG in this patient. Above finding has provided a basis for the diagnosis and genetic counseling for this family.

Effects of alternating food restriction on blood glucose, body mass index and blood lipids in overweight or obesity patients with type 2 diabetes mellitus / 中国医师进修杂志

Objective:To explore the effect of alternating food restriction on blood glucose, body mass index (BMI) and blood lipids in overweight or obesity patients with type 2 diabetes mellitus.Methods:A prospective cohort study was used. Three hundred overweight or obesity type 2 diabetes mellitus patients with stable blood glucose control from December 2021 to February 2022 in Nanxiang Hospital, Jiading District of Shanghai City were selected. The patients were divided into alternating food restriction group (adopting alternating food restriction therapy, giving balanced meal plates, reducing 30% of calories intake every other day), low carbohydrate high protein group (adopting low carbohydrate and high protein therapy, giving low carbohydrate and high protein reduction meal plates, reducing 15% of calories intake every day) and balanced diet group (adopting balanced diet therapy, giving balanced meal plates) by random digits table method with 100 cases each. All three groups received intervention treatment for 6 months. The height and body mass before intervention and the end of intervention and 6 months after intervention were measured, and the BMI was calculated. The levels of glycosylated hemoglobin (HbA 1c), fasting blood glucose (FBG), 2 h postprandial blood glucose (2 h PBG), triacylglycerol (TG), total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) were measured. Results:At the end, 280 cases were completed the study. There were 90 cases in the alternating food restriction group, 90 cases in the low carbohydrate high protein group, and 100 cases in the balanced diet group. There were no statistical differences in HbA 1c, FBG, 2 h PBG, BMI, TG, TC and LDL-C before intervention among the three groups ( P>0.05). At the end of the intervention, the HbA 1c and FBG in alternating food restriction group and low carbohydrate high protein group were significantly lower than those in balanced diet group: (6.50 ± 0.39)% and (6.67 ± 0.30)% vs. (6.79 ± 0.32)%, (6.47 ± 0.61) and (6.80 ± 0.30) mmol/L vs. (6.94 ± 0.37) mmol/L, the indexes in alternating food restriction group were significantly lower than those in low carbohydrate high protein group, and there were statistical difference ( P<0.05); the 2 h PBG and BMI in alternating food restriction group and the low carbohydrate high protein group were significantly lower than those in balanced diet group: (8.83 ± 0.63) and (8.81 ± 0.70) mmol/L vs. (9.45 ± 0.85) mmol/L, (25.99 ± 2.13) and (26.53 ± 2.16) kg/m 2 vs. (27.24 ± 2.24) kg/m 2, and there were statistical differences ( P<0.05), there were no statistical differences in 2 h PBG and BMI between alternating food restriction group and the low carbohydrate high protein group ( P>0.05). Six months after intervention, the HbA 1c, 2 h PBG and BMI in alternating food restriction group were significantly lower than those in low carbohydrate high protein group and balanced diet group: (6.62 ± 0.29)% vs. (6.79 ± 0.19)% and (6.84 ± 0.23)%, (9.21 ± 0.53) mmol/L vs. (9.48 ± 0.66) and (9.55 ± 0.51) mmol/L, (25.60 ± 1.67) kg/m 2 vs. (27.26 ± 2.42) and (27.79 ± 2.49) kg/m 2, and there were statistical differences ( P<0.05), there were no statistical differences in HbA 1c, 2 h PBG and BMI between low carbohydrate high protein group and balanced diet group ( P>0.05). At the end of intervention and 6 months after intervention, there were statistical differences in TG, TC and LDL-C among the three groups ( P<0.05); among them, the TG in alternating food restriction group was significantly lower than that in low carbohydrate high protein group and the balanced diet group: (1.67 ± 0.70) mmol/L vs. (1.99 ± 0.89) and (2.49 ± 0.94) mmol/L, (1.70 ± 0.71) mmol/L vs. (2.04 ± 0.96) and (2.53 ± 1.08) mmol/L, and there were statistical differences ( P<0.05), there was no statistical difference in TG between the low carbohydrate high protein group and balanced diet group ( P>0.05). Conclusions:The alternating food restriction therapy in overweight or obesity patients with type 2 diabetes mellitus can not only reduce blood glucose, improve blood lipids, but also reduce BMI, and the overall effect is better than that of low carbohydrate high protein therapy.