Objective To characterize the properties of Hepa1-6-derived microparticles (Hepa1-6-MPs), investigate their stimulatory effects on dendritic cells (DCs) and their cellular uptake pathways, and explore the specific cytotoxic effects of CD8⁺ T cells induced by Hepa1-6-MP-loaded DCs on hepatoma cell lines, with the aim of developing a novel immunotherapeutic model for hepatocellular carcinoma (HCC). Methods The isolated Hepa1-6-MPs were identified using Western blotting, transmission electron microscopy (TEM) and dynamic light scattering (DLS). Flow cytometry was used to assess the uptake pathways of Hepa1-6-MPs by DCs. Subsequently, enzyme-linked immunosorbent assay (ELISA) was used to measure the effects of Hepa1-6-MP-loaded DCs on the release levels of tumor necrosis factor α(TNF-α) and interferon γ(IFN-γ) into the supernatant of CD8⁺ T cells. Lactate dehydrogenase (LDH) tests were conducted to evaluate the cytotoxic effects of CD8⁺ T cells stimulated by Hepa1-6-MP-loaded DCs on hepatoma cells. Results The morphology, size and protein markers of Hepa1-6-MPs met the established criteria. Hepa1-6-MPs enhanced the expression of DC maturation markers CD80 and CD86, and were internalized by DCs via clathrin-mediated endocytosis and phagocytosis pathways. Subsequently, Hepa1-6-MP-loaded DCs stimulated CD8⁺ T cells to release high levels of TNF-α and IFN-γ, which induced their specific cytotoxicity against HCC cells. Conclusion These findings suggest that Hepa1-6-MP-loaded DCs may be a promising HCC immunotherapeutic tool.
Carcinoma, Hepatocellular/therapy* ; Liver Neoplasms/therapy* ; Dendritic Cells/immunology* ; Humans ; Cancer Vaccines/immunology* ; CD8-Positive T-Lymphocytes/immunology* ; Cell Line, Tumor ; Tumor Necrosis Factor-alpha/immunology* ; Interferon-gamma/immunology* ; Cell-Derived Microparticles/immunology* ; Animals

Idiopathic gingival fibromatosis is a rare, benign condition of unknown etiology characterized by extensive gingival overgrowth. This case reports a severe skeletal Class Ⅱ adult female patient with idiopathic gingival fibromatosis. The patient underwent multidisciplinary treatment involving periodontics, orthodontics, and orthognathic surgery, resulting in remarkable crown height elongation, substantial improvements in occlusal function and aesthetics, and stable long-term follow-up outcomes. This case provides a reference for future clinical practice.
Humans ; Female ; Malocclusion, Angle Class II/complications* ; Fibromatosis, Gingival/complications* ; Adult ; Orthodontics, Corrective ; Orthognathic Surgical Procedures

AIM:To establish an optimal protocol for isolating and extracting mouse hepatocellular carcinoma H22 cell-detrived microparticles(H22-MPs),evaluate their stimulating effect on dendritic cells(DC),and investigate the cytotoxic capacity of CD8+T cells induced by H22-MPs on hepatocellular carcinoma cells.METHODS:The H22-MPs were extracted by high-speed differential centrifugation,and the morphology of H22-MPs was observed by transmis-sion electron microscopy.The particle size distribution of H22-MPs was detected by dynamic light scattering,and the ex-pression of vesicle surface markers was detected by Western blot.The protein components and pathways involved by MPs were analyzed by labeling free quantitative proteomics combining with bioinformatics GO and KEGG enrichment methods.The effect of MPs antigen on the expression of CD11c,a mature marker of DC,was detected by laser confocal microscopy.Next,CD8+T cells were sorted by magnetic beads,and the secretion of tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ)and interleukin-2(IL-2)in the supernatant of CD8+T cells stimulated by DC loaded with H22-MPs was detected by ELISA.Lactate dehydrogenase(LDH)test was conducted to evaluate the specific killing effect of CD8+T cells induced by DC loaded with H22-MPs on hepatocellular carcinoma cells.RESULTS:The results showed that the morphology,size and vesicle markers of H22-MPs met the requirements.Most of the H22-MPs protein antigens were derived from its parent cell H22 and coordinated various signal transduction pathways in the cells.Subsequently,it was detected that H22-MPs increased the expression of CD11c on the surface of DC,and H22-MPs were mainly localized in the lysosomes of DC.In addition,DC loaded with H22-MPs stimulated CD8+T to release high levels of TNF-α,IFN-γ and IL-2,and it also pro-moted CD8+T to overexpress CD69,thus inducing CD8+T to killhepatocellular carcinoma cells specifically.CONCLU-SION:H22-MPs promote the maturation of DC,and the mature DC may induce CD8+T cells to play an anti-hepatocellu-lar carcinoma immune response by promoting the activation of CD8+T cells.

AIM:To establish an optimal protocol for isolating and extracting mouse hepatocellular carcinoma H22 cell-detrived microparticles(H22-MPs),evaluate their stimulating effect on dendritic cells(DC),and investigate the cytotoxic capacity of CD8+T cells induced by H22-MPs on hepatocellular carcinoma cells.METHODS:The H22-MPs were extracted by high-speed differential centrifugation,and the morphology of H22-MPs was observed by transmis-sion electron microscopy.The particle size distribution of H22-MPs was detected by dynamic light scattering,and the ex-pression of vesicle surface markers was detected by Western blot.The protein components and pathways involved by MPs were analyzed by labeling free quantitative proteomics combining with bioinformatics GO and KEGG enrichment methods.The effect of MPs antigen on the expression of CD11c,a mature marker of DC,was detected by laser confocal microscopy.Next,CD8+T cells were sorted by magnetic beads,and the secretion of tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ)and interleukin-2(IL-2)in the supernatant of CD8+T cells stimulated by DC loaded with H22-MPs was detected by ELISA.Lactate dehydrogenase(LDH)test was conducted to evaluate the specific killing effect of CD8+T cells induced by DC loaded with H22-MPs on hepatocellular carcinoma cells.RESULTS:The results showed that the morphology,size and vesicle markers of H22-MPs met the requirements.Most of the H22-MPs protein antigens were derived from its parent cell H22 and coordinated various signal transduction pathways in the cells.Subsequently,it was detected that H22-MPs increased the expression of CD11c on the surface of DC,and H22-MPs were mainly localized in the lysosomes of DC.In addition,DC loaded with H22-MPs stimulated CD8+T to release high levels of TNF-α,IFN-γ and IL-2,and it also pro-moted CD8+T to overexpress CD69,thus inducing CD8+T to killhepatocellular carcinoma cells specifically.CONCLU-SION:H22-MPs promote the maturation of DC,and the mature DC may induce CD8+T cells to play an anti-hepatocellu-lar carcinoma immune response by promoting the activation of CD8+T cells.

Objective:To investigate the value of early tight junction protein Claudin-5 levels in predicting the severity of acute pancreatitis (AP).Methods:A prospective observational study was conducted, including patients diagnosed with AP and admitted to the Northern Jiangsu People's Hospital from December 1, 2021 to November 30, 2022. Eligible healthy volunteers were randomly selected to serve as healthy controls during the same period. Patients were classified into mild acute pancreatitis (MAP) group, moderate-severe acute pancreatitis (MSAP) group, and severe acute pancreatitis (SAP) group based on the 2012 Atlanta classification criteria. Patients with SAP were then divided into three subgroups of 1, 3, and 7 days based on the duration of hospitalization. Baseline data, such as gender, age, underlying disease, and probable etiology, was collected from all enrolled individuals. The enzyme-linked immunosorbent assay (ELISA) was employed to detect serum Claudin-5 levels in each cohort of enrollees. Data on additional serologic indicators, including hematocrit (HCT), albumin (Alb), serum Ca 2+, C-reactive protein (CRP), and procalcitonin (PCT) levels, were obtained via the in-hospital test query system in each group of patients with AP. The modified Marshall score (mMarshall), modified CT severity index (mCTSI) score, bedside severity index of severity in acute pancreatitis (BISAP) score, and acute physiology and chronic health evaluation Ⅱ(APACHEⅡ) were recorded for each group of patients with AP. Differences in the above indicators between groups were analyzed and compared. Spearman's correlation method was employed to examine the relationship between Claudin-5 levels and each influential factor. The receiver operator characteristic curve (ROC curve) was plotted to analyze the predictive value of each influencing factor on SAP. Ridge regression was used to screen for independent risk factors for SAP. Results:A total of 109 patients with AP were enrolled, comprising 66 in the MAP group, 15 in the MSAP group, and 28 in the SAP group. Additionally, 27 healthy volunteers were enrolled as the healthy control group. No statistically significant differences were observed in gender and age among the enrolled groups, and no statistically significant differences were identified among the three groups of patients with AP in terms of underlying disease and etiologic composition. As the disease progressed, serum Claudin-5 levels exhibited a notable increase across all AP patient groups, and they were all significantly higher than those in the healthy control group [ng/L: 888.58 (574.52, 1 141.59), 3 749.02 (2 784.93, 5 789.92), 4 667.81 (3 935.21, 7 315.66) vs. 291.13 (250.19, 314.75), all P < 0.05]. Subgroup analyses showed that as the disease duration prolonged, patients in the SAP group exhibited a notable decline in Claudin-5 levels at 3 days post-admission, compared with those at 1 day post-admission [ng/L: 2 052.59 (1 089.43, 4 006.47) vs. 4 667.81 (3 935.21, 7 315.66), P < 0.05]. Spearman correlation analysis showed that serum Claudin-5 levels in patients with AP were significantly positively correlated with CRP, PCT, HCT, and mMarshall, mCTSI, and BISAP scores ( r values were 0.570, 0.525, 0.323, 0.774, 0.670, 0.652, all P < 0.001), and significantly negatively correlated with Alb ( r = -0.394, P < 0.001). A significant trend was observed in patients with AP, with an increase of HCT levels and a decrease of Alb levels as the disease progressed (both P < 0.05). An improvement of aforementioned phenomena was observed in patients with SAP following treatment, indirectly indicating that serum Claudin-5 level was a positive indicator of vascular leakage. ROC curve analysis showed that serum Claudin-5 levels in patients with AP exhibited the highest accuracy for early prediction of SAP, with the area under the ROC curve (AUC) of 0.948. When serum Claudin-5 levels ≥2 997 ng/L, the sensitivity for early screening for SAP was 100% and the specificity was 88.89%. Multifactorial ridge regression analysis showed that serum Claudin-5 level, PCT and APACHEⅡscore could be used as independent risk factors for early prediction of SAP (all P < 0.05). Conclusion:Serum Claudin-5 levels facilitate early prediction of SAP and are strongly associated with inflammatory response and vascular leakage.

Objective To explore the value of synchrotron radiation micro-CT(SR-pCT)and automatic marker watershed image segmentation for quantifying three-dimensional local bone tissue and blood vessels in mouse models of breast cancer tibia metastasis.Methods Mouse models with breast cancer bone metastasis in tibia were established and underwent oral Vatalanib,injection of Bevacizumab,combination of Vatalanib or Bevacizumab,or injection of placebo,respectively.On the 8th day(time point T1)and the 22nd day(time point T2)after injection of tumor cells,the tibia bone was scanned using combined vascular contrast injection and SR-pCT,and high-resolution simultaneous imaging of local bone tissue and vascular network were achieved.Then bone tissue and vessels on images were segmented using automatic marker watershed image-segmentation method,global threshold method and region growing method,respectively,and the quality of segmentations were assessed with Dice similarity coefficient(DSC).The anatomical quantitative parameters of tibia with breast cancer metastasis were obtained,including the fraction of bone volume in total volume(BV/TV),fraction of vessel volume in total volume(VV/TV),the mean thickness of bone(B.Th)and the mean thickness of vessel(V.Th).The anatomical quantitative parameters were compared among different treatments at the same time point,also between different time points under the same treatment.Results DSC for segmentation of local tibia bone tissue and vascular structures based on automatic marker watershed image segmentation method were all greater than 0.95,higher than that of other methods(all P＜0.05).No significant difference of anatomical quantitative parameters was found under different treatments at T1(all P＞0.05),and V.Th under the combined treatment were lower than that under other treatments at T2(all P＜0.05).Compared with T1,BV/TV and B.Th at T2 increased while VV/TV at T2 decreased in tibia metastasis under combination treatment(all P＜0.05),BV/TV,B.Th and V.Th at T2 increased under Vatalanib or Bevacizumab treatment(all P＜0.05),and BV/TV and B.Th at T2 increased under placebo(both P＜0.05).Conclusion The combination of SR-pCT and automatic marker watershed method could be used to initially quantify three-dimensional structures of local bone tissue and blood vessels in mouse models of breast cancer tibia metastasis.

OBJECTIVE: To investigate the clinical and genetic characteristics of a patient with STISS syndrome due to variant of PSMD12 gene. METHODS: Clinical data and result of genetic testing of a patient who was admitted to Shanghai Children's Medical Center, Shanghai Jiaotong University School of Medicine on October 4, 2020 were analyzed, together with a review of relevant literature. RESULTS: The patient was found to harbor a heterozygous c.601C>T (p.Arg201*) nonsense variant of the PSMD12 gene, which was unreported previously. Clinically, the height of the patient has differed significantly from reported in the literature. An extremely rare case of STISS syndrome due to variant of the PSMD12 gene has been diagnosed. CONCLUSION Whether the severely short stature is part of the clinical spectrum for PSMD12 gene variants needs to be further explored, and the efficacy and safety of growth hormone therapy has yet to be determined.
Child ; Humans ; China ; Dwarfism ; Genetic Testing ; Heterozygote ; Syndrome

[摘 要] 目的：探讨幽门螺杆菌（Helicobacter pylori, Hp）感染对胃癌细胞共济失调毛细血管扩张突变（ataxia-telangiectasia mutated，ATM）基因表达的影响及其临床意义。方法：从TCGA数据库中获取胃癌相关RNAseq数据，比较ATM基因的表达差异，分析ATM表达与患者临床病理参数的相关性及预后价值，用Kaplan-Meier法进行生存分析，LinkedOmics数据库分析ATM相关基因，用R语言进行GO、KEGG富集分析。选用2019年3月至2019年12月贵州医科大学附属医院12例手术切除的胃癌及癌旁组织标本，以及胃癌细胞系AGS和BGC823，用感染复数40∶1的Hp GZ7菌感染细胞，用免疫组织化学染色法检测胃癌组织中ATM蛋白的表达，qPCR法检测胃癌组织和细胞中ATM mRNA的表达。结果：TCGA数据显示胃癌和Hp感染胃癌组织中ATM miRNA表达水平均显著高于癌旁组织（均P<0.01）；胃癌组织中ATM miRNA表达与患者的T分期、AJCC分期等病理参数呈正相关（均P<0.05），ATM高表达时生存率显著降低（P<0.05）。实验检测显示，胃癌组织标本中ATM蛋白的表达水平明显高于癌旁组织（P<0.01）；Hp感染胃癌细胞中ATM miRNA表达水平显著高于未感染胃癌细胞（P<0.01）。胃癌中ATM基因与NPAT等12 461个基因呈正相关（P<0.05），与MIF等7 764个基因呈负相关（P<0.05）。GO、KEGG富集分析显示，ATM富集到DNA修复复合体、癌症中的转录失调等信号通路。结论：ATM基因在胃癌组织中高表达，患者生存率随表达水平的增高而降低，其与患者的T分期、AJCC分期等病理参数相关，且Hp感染引起ATM表达水平升高可能是Hp引起胃癌的原因之一。

OBJECTIVE：To shar e the experienc e of pharmaceutical care in Wuhan Jinyintan Hospital （herein after refers to “our hospital ”）under the condition of novel coronavirus pneumonia （COVID-19）epidemic，and to provide reference for other hospitals to deal with public health emergencies. METHODS ：The situation of pharmaceutical care in our hospital under the condition of COVID- 19 epidemic was summarized and shared ，including the epidemic prevention and control management （regional division ，disinfection management ，pharmacy personnel training ），supply of drugs and disinfection products ，the monitoring and education of rational drug use by information technology. RESULTS ：The pharmacy department of our hospital divided the activity scope into clean area ，potential pollution risk area ，semi pollution area ，and implement different disinfection management. All pharmacists received training ，involving personal health protection ，prevention and control knowledge of COVID-19，health status monitoring ，etc. For supply and guarantee of drugs and disinfectants ，the epidemic drug list of our hospital was formulated ，drugs and disinfectants were purchased accurately and stored in a standardized way. 24 h response telephone was set up in the clinical pharmacy room to receive consultation from clinicians on drug use at any time. The drugs mentioned in the COVID- 19 diagnosis scheme were compared in terms of the mechanism of action and the medication of special populations to form a tablet ，so as to help clinical rational choice treatment drug. CONCLUSIONS ：The pharmaceutical care in the designated hospital of COVID- 19 is a professional and complicated work ，involving a wide range of aspects. Pharmacy department must respond actively and adjust the strategy in time so as to play an important role in improving the ability of medical treatment.

Objective To approach the predictive value of continuous monitoring end-tidal carbon dioxide partial pressure (PETCO2) on the outcome of in-hospital cardiopulmonary resuscitation (CPR), and explored the indicators of termination of resuscitation. Methods A secondary analysis of a multicenter observational study data was conducted. The screening aim was adult non-traumatic in-hospital CPR patients whose PETCO2were recorded within 30 minutes of CPR. Clinical information was reviewed. The mean PETCO2in restoration of spontaneous circulation (ROSC) and non-ROSC patients was recorded. The outcome of CPR was continuously assessed by PETCO2≤ 10 mmHg (1 mmHg = 0.133 kPa) for 1, 3, 5, 8, 10 minutes. Receiver operating characteristic (ROC) curve was plotted, and the predictive value of PETCO2≤ 10 mmHg for different duration on the outcome of CPR was evaluated. Results A total of 467 recovery patients, including 419 patients with complete recovery were screened. Patients who were out-of-hospital resuscitation, non-adults, traumatic injury, had no PETCO2value, PETCO2value failed to explained the clinical conditions, or patients had not monitored PETCO2within 30 minutes of resuscitation were excluded, and finally 120 adult patients with non-traumatic in-hospital resuscitation were enrolled in the analysis. The mean PETCO2in 50 patients with ROSC was significantly higher than that of 70 non-ROSC patients [mmHg: 17 (11, 27) vs. 9 (6, 16), P < 0.01]. ROC curve analysis showed that the area under ROC curve (AUC) of PETCO2during the resuscitation for predicting recovery outcome was 0.712 [95% confidence interval (95%CI) = 0.689-0.735]; when the cut-off was 10.5 mmHg, the sensitivity was 57.8%, and the specificity was 78.0%, the positive predictive value (PPV) was 84.6%, and negative predictive value (NPV) was 46.9%. The duration of PETCO2≤ 10 mmHg was used for further analysis, which showed that with PETCO2≤10 mmHg in duration, the prediction of the sensitivity of the patients failed to recover decreased from 58.2% to 28.2%, but specificity increased from 39.4% to 100%; PPV increased from 40% to 100%, and NPV decreased from 57.5% to 34.2%. Conclusion For adult non-traumatic in-hospital CPR patients, continuous 10 minutes PETCO2≤10 mmHg may be an indicate of termination of CPR.