To investigate the effect of silencing DJ-1 on xenografted human laryngeal squamous cell carcinoma (LSCC) Hep-2 cells in nude mice.Xenograft model of human LSCC was established by subcutaneous transplantation of Hep-2 cells in 24 nude mice. The LSCC-bearing nude mice were randomly divided into 3 groups (=8 in each):DJ-1 siRNA low dose group and DJ-1 siRNA high dose group were injected in tumors with 20 μg of DJ-1 siRNA or 40 μg of DJ-1 siRNA in 50 μL, respectively; control group was injected with 5% glucose solution in 50 μL, twice a week for 3 weeks. The weight and size of tumors were measured before injection. The animals were sacrificed 48 h after the final treatment, and the tumors were harvested and weighed. The apoptosis and proliferation of tumor cells were determined; the expressions of Caspase-3 and Ki-67 in tumor specimens were detected with immunohistochemistry. The expression of DJ-1, PTEN, survivin mRNA and protein in tumor tissues were detected by RT-PCR and Western blotting, respectively.Tumor weight in low dose group[(0.66±0.15)g] and high dose group[(0.48±0.11)g] were significantly lower than that in control group[(0.83±0.16)g, all<0.05]. The inhibition rates of low dose group and high dose group were (20.48±0.18)% and (42.16±0.13)%, respectively. Immunohistochemistry showed that the expression of Caspase-3 was increased and Ki-67 was reduced in tumor specimens, compared with the control group (all<0.05). RT-PCR and Western blot results showed that in low dose group and high dose group the mRNA and protein expression of DJ-1 and survivin significantly decreased (all<0.05), while PTEN mRNA and protein content increased (all<0.05).High dose DJ-1 siRNA can inhibit the tumor growth in human LSCC xenograft nude mouse model, which indicates that down-regulating DJ-1 and survivin, and up-regulating PTEN expression may lead to blockage of PI3K-PKB/Akt signaling pathway and promoting tumor cell apoptosis.
Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; genetics ; Carcinoma, Squamous Cell ; chemistry ; genetics ; physiopathology ; Caspase 3 ; analysis ; drug effects ; Cell Line, Tumor ; chemistry ; drug effects ; physiology ; transplantation ; Cell Proliferation ; drug effects ; Down-Regulation ; Gene Expression Regulation ; drug effects ; genetics ; physiology ; Head and Neck Neoplasms ; chemistry ; genetics ; physiopathology ; Heterografts ; drug effects ; physiology ; Humans ; Inhibitor of Apoptosis Proteins ; analysis ; drug effects ; Ki-67 Antigen ; analysis ; drug effects ; Laryngeal Neoplasms ; chemistry ; genetics ; physiopathology ; Mice, Nude ; PTEN Phosphohydrolase ; analysis ; drug effects ; Phosphatidylinositol 3-Kinases ; drug effects ; Protein Deglycase DJ-1 ; pharmacology ; Proto-Oncogene Proteins c-akt ; drug effects ; RNA Interference ; physiology ; RNA, Messenger ; pharmacology ; RNA, Small Interfering ; physiology ; Signal Transduction ; drug effects ; genetics ; physiology

Objectives To explore if the rat bone marrow mesenchymal stem cells (BMSCs) modified by human heme oxygenase 1 (hHO-1) gene combined with GATA-4 gene may promote the ability of anti-apoptosis and myocardial transdifferentiation in vitro in hypoxia ischemic environment.Methods The rat BMSCs were isolated and cultured by whole bone marrow adherence and identified in vitro,and then were transfected with recombinant adenovirus;Western blotting was used to determinate the optimal time of gene expression;the genetically modified BMSCs were taken to hypoxia serum-free conditions simulating ischemia hypoxia microenvironment in vivo;CCK-8 kit and trypan blue staining were performed to detect the 12,24,48 and 72h survival rates in hypoxia ischemia respectively;flow cytometry was used to detect the apoptosis of BMSCs in hypoxia ischemia for 24h.The cardiomyocyte-specific cardiac troponin Ⅰ (cTnI) was detected by Western blotting and cellular immunofluorescence.Results The 12,24,48 and 72h survival rates were higher in hHO-1+GATA-4 group cultured in ischemia and hypoxia condition than in hHO-1 group (P＜0.05) and GATA-4 group (P＜0.05).After 24h cultivation in ischemia hypoxia condition,the apoptotic rates were lower in hHO-1+GATA-4 group than in hHO-1 group (P＜0.05) and GATA-4 group (P＜0.05).No significant difference existed in cTnI expressions between GATA-4 group and hHO-1+GATA-4 group.Conclusion Compared with transfection of hHO-1 or GATA-4 single gene,hHO-1 combined with GATA-4 transduction can significantly increase the survival rate of BMSCs in hypoxia ischemic condition,but myocardial transdifferentiation does not increase significantly.

OBJECTIVES: To explore the species, quantity and distribution of diatoms in Ningbo three-river watershed during summer and to provide scientific basis for forensic examination of drowning cases in the waters of Ningbo. METHODS: Water samples were collected in July and August of 2015. Fourteen water sampling points were selected from the Yao River, the Fenghua River and the Yong River. The morphological features of diatom species and dominant diatoms were distinguished by microscope. RESULTS: A total of 16 species of diatoms were detected in the Yao River, the Fenghua River and the Yong River. Melosira was the dominant species in the Yao River, and the quantity and richness were higher than in other rivers. The richness of Cyclotella in the Yong River was higher than in other rivers. The richness of Pinnularia and Licmophora were higher in the Fenghua River than in the Yao River and the Yong River. CONCLUSIONS The species and proportion of diatom is different in each river. Database of the species and relative composition for the diatoms in corresponding river is established, which may provide data support for forensic examination of drowning cases in Ningbo three-river watershed.
China ; Diatoms/classification* ; Drowning ; Rivers ; Seasons

【Objective】 To investigate the impact of human leukocyte antigen (HLA) functional epitope mismatch (EM) on the efficacy of platelet transfusion in patients with hematological diseases. 【Methods】 HLA genotyping was performed on platelet donors and patients with hematological diseases who applied for platelet serological cross-matching and HLA antigen matching from June 2021 to June 2023 by PCR-SBT method. HLA platelet matching was based on the principle of CREG to select donors for patients. HLA Matchmaker 4.0 software was used to analyze donor-recipient HLA EM information. The expression level and gene distribution of related HLA functional epitope (Eplet) were searched from the international HLA Epitope registry website (www.Epregistry.com.br). Retrospective analysis was conducted on clinical platelet transfusion data to evaluate the impact of HLA EM on platelet transfusion effectiveness. 【Results】 Platelet transfusion efficacy showed no correlation with gender and age, but it was associated with platelet matching strategy. When the total number of HLA EMs was less than 20, a lower total number of donor-recipient HLA EMs resulted in higher platelet transfusion efficiency (χ²=19.311, P=0.001) and higher average value of 24 h corrected count increment (CCI)(F=7.737, P<0.001). The total number of donor-recipient HLA EMs had negative correlation with actual 24 h CCI (Rho=-0.322, P<0.001). Further statistical analysis revealed that 17 Eplets were related to the effectiveness of platelet transfusion. The locus distribution of 17 Eplets might be unique to HLA-A (17.6%) or -B (64.7%) or shared between HLA-A and -B (17.6%), and its expression may be high (58.8%) or intermediate (41.2%). 【Conclusion】 The total number of donor-recipient HLA EMs is a crucial factor influencing platelet transfusion effectiveness, and several HLA Eplets associated with this effectiveness have been identified.

OBJECTIVETo explore the molecular basis of 4 cases with weak D variant of Rh blood type.

METHODSRoutine serological testing was applied to determine the D, C, c, E and e antigens of the Rh blood group. The D antigen was further detected with an indirect antiglobulin test. RHD zygosity was detected by sequence-specific primer PCR method. All exons and flanking intron regions of the RHD gene were sequenced.

RESULTSThe samples were determined as weak D phenotype by serological testing. DNA sequencing showed that the 4 cases were heterozygous for 17C>T mutation in exon 1, 29G>C mutation in exon 1, 1212C>A mutation in exon 9, and IVS4+5G>A mutation in intron 4 of the RHD gene, respectively. According to the rule of Rhesus Base Nomenclature, the 4 samples were respectively named as weak D type 31, weak D type 71, weak D type 72, and weak D type 82.

CONCLUSIONSerological and molecular testing for the weak D can facilitate in-depth understanding of its immunology and genetics, and provide guidance for clinical blood transfusion and prevention of hemolytic disease in newborns.

Acute-On-Chronic Liver Failure ; diagnosis ; virology ; Antiviral Agents ; therapeutic use ; Hepatitis B, Chronic ; drug therapy ; Humans ; Nucleosides ; therapeutic use ; Nucleotides ; therapeutic use ; Prognosis

Objective: To analyze the classification and functions of cell subsets in laryngeal carcinoma and metastatic lymph nodes, and to explore the evolution trajectory of epithelial cells to tumor cells. Methods: Single-cell RNA sequencing was performed on 5 cases of laryngeal cancer, matched metastatic lymph nodes and 3 normal tissues. Patients were admitted to Ningbo Medical Center Lihuili Hospital from October 22, 2019 to December 16, all patients were male, aged 53-70 years old. Cell subsets of the above-mentioned tissues were analyzed by the Seurat, and the biological functions of cell subpopulation were investigated by functional enrichment analysis. Malignant epithelial cells were identified using copy number variation (CNV). The evolutionary trajectory of epithelial cells to cancer cells was analyzed by cell trajectory analysis, and cancerous transitional cells were identified. The highly expressed genes in transitional cells were analyzed by the FindAllMarker of the Seurat and verified by immunohistochemistry. Results: A total of 66 969 high-quality cells were obtained in 9 major clusters: epithelial cells, T cells, B cells, fibroblasts, endothelial cells, myeloid cells, mast cells, plasmacytoid dendritic cells and nerve cells. The first 5 cell clusters were divided into 8, 6, 4, 3 and 2 subgroups, respectively. Four epithelial cell subsets (C0, C1, C2 and C5) were derived from tumor tissues and metastatic lymph nodes, and had high levels of CNV and tumor cell content. Cell trajectory analysis showed that the evolution trajectory of epithelial cells was from normal epithelial subpopulation C4 to early cancerous cell population C0, which differentiated into three major malignant cell subsets C1, C3, and C5. Epithelial cell C0 may represent the transitional cell population of carcinogenesis, and were enriched in biological processes such as epithelial-mesenchymal transformation and angiogenesis. C0 highly expressed sulforaphane (SFN) which may be related to the occurrence and development of cancer. Immunohistochemistry confirmed that SFN was highly expressed in tumor tissues and metastatic lymph nodes compared with paracancerous tissues. Conclusion: Single-cell sequencing may be used to elucidate the diversity of cells and functions in laryngeal carcinoma tissues and metastatic lymph nodes, and cell population C0 plays a key role in the evolution of cells.
Aged ; Carcinoma, Squamous Cell/pathology* ; DNA Copy Number Variations ; Endothelial Cells/pathology* ; Humans ; Laryngeal Neoplasms/pathology* ; Lymph Nodes/pathology* ; Male ; Middle Aged

case of schwannoma with hemorrhagic cystic degeneration in the right upper mediastinum was admitted to the Affiliated Hospital of Ningbo University in July 2010. The patient shows symptoms of cough and shortness of breath. He received video-assisted thoracoscopic resection of right upper mediastinal mass. This disease displayed different symptoms depending on tumor size and location.
Humans ; Male ; Mediastinum ; pathology ; Neurilemmoma ; diagnosis ; pathology

Nonalcoholic fatty liver disease(NAFLD)has developed into the most common chronic liver disease and can lead to liver cancer.Our laboratory previously developed a novel prescription for NAFLD,"Eight Zhes Decoction"(EZD),which has shown good curative effects in clinical practice.However,the pharmaco-dynamic material basis and mechanism have not yet been revealed.A strategy integrating lipidomics,network pharmacology and pharmacokinetics was used to reveal the active components and mecha-nisms of EZD against NAFLD.The histopathological results showed that EZD attenuated the degrees of collagen deposition and steatosis in the livers of nonalcoholic steatofibrosis model mice.Furthermore,glycerophospholipid metabolism,arachidonic acid metabolism,glycerolipid metabolism and linoleic acid metabolism with phospholipase A2 group IVA(PLA2G4A)and cytochrome P450 as the core targets and 12,13-cis-epoxyoctadecenoic acid,12(S)-hydroxyeicosatetraenoic acid,leukotriene B4,prostaglandin E2,phosphatidylcholines(PCs)and triacylglycerols(TGs)as the main lipids were found to be involved in the treatment of NAFLD by EZD.Importantly,naringenin,artemetin,canadine,and bicuculline were iden-tified as the active ingredients of EZD against NAFLD;in particular,naringenin reduces PC consumption by inhibiting the expression of PLA2G4A and thus promotes sufficient synthesis of very-low-density lipoprotein to transport excess TGs in the liver.This research provides valuable data and theoretical support for the application of EZD against NAFLD.

Objective To analyze the effect of Xiaozhi Decoction in the treatment of hyperlipidemiain different classification of physical constitution in TCM.Methods We screened 206 patients with Hyperlipidemia in our hospital and had been treated with drugs during May 2020 to March 2023.Totally 103 patients in the TCM group were treated with Xiaozhi Decoction,103 patients in the western medicine group were treated with atorvastatin.Selected total cholesterol(TC),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C)and liver transaminase before and after a period of treatment.The non-high-density lipoprotein cholesterol(non-HDL-C)will be calculated,too.Results In the Phlegm-Dampness constitution,TC,TG,LDL-C and non-HDL-C decreased significantly all in the TCM group(P<0.05);TC,TG and non-HDL-C decreased significantly all in the western medicine group(P<0.05).The TCM group is superior to the western medicine group in TC,LDL-C,non-HDL-C(P<0.05).In the Qi-Deficiency constitution,TC,TG,LDL-C and non-HDL-C decreased significantly all in the TCM group(P<0.05);TC,LDL-C and non-HDL-C decreased significantly all in the western medicine group(P<0.05).The TCM group is superior to the western medicine group in non-HDL-C(P<0.05).In the Blood-Stasis constitution,TC,TG,LDL-C and non-HDL-C decreased significantly all in the western medicine group only(P<0.05).The western medicine group is superior to the TCM group in TC,LDL-C,non-HDL-C(P<0.05).In the Yin-Yang Harmony constitution,TC and non-HDL-C decreased significantly both in the TCM group(P<0.05).TC,LDL-C and non-HDL-C decreased significantly all in the western medicine group(P<0.05).The western medicine group is superior to the TCM group in TC,LDL-C,non-HDL-C(P<0.05).Conclusion Xiaozhi Decoction is superior to the the atorvastatin in the treatment of hyperlipidemia for the Phlegm-Dampness constitution and Qi-deficiency constitution groups.But it is not superior to the atorvastatin in the treatment of hyperlipidemia for the Blood-Stasis constitution and Yin-Yang Harmony constitution groups.