Table of Content

05 May 2024, Volume 44 Issue 5

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Specialists Consensus

Consensus on endocrine management for children and adolescents with craniopharyngioma surgeries

Growth and Development and Gonadal Diseases Committee of Chinese Aging Well Association

2024, 44(5):  585-598.  doi:10.16352/j.issn.1001-6325.2024.05.0585

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Craniopharyngioma is a rare benign tumor mainly occurs in the hypothalamus region of children. Patients may have a long survival after surgery. Tumors or operations may lead to hypothalamic syndrome, hypopituitarism, obesity, and a disturbed electrolytes metabolism. Multidisciplinary medical teams are required to conduct a long-term hormonal therapy and follow-up management. This consensus is aiming for providinge recommendations for perioperative management, obesity, hormonal replacement therapy, linear growth promotion and pubertal development for pediatric patients.

Original Articles

Glioma conditioned medium promotes glycolysis and phenotypic transformation in macrophages

ZHANG Diya, LI Yunfan, GUO Hongjiang, QIU Jiaxing, WANG Yucheng, JU Rui, GUO Lei

2024, 44(5):  599-605.  doi:10.16352/j.issn.1001-6325.2024.05.0599

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Objective To explore the effects and mechanisms involved in glioma conditioned medium for inducing glycolysis and oncogenic mediators in macrophages. Methods Bone marrow derived macrophage (BMDM) induced by hypoxia and mouse glioma cell line GL261 conditioned medium (GCM)(GCM-BMDM) were used as a model. The mRNA level of M1 and M2 oncogenic mediators, hypoxia-inducible factor-1α (HIF-1α) and glycolysis associated genes in GCM-BMDM were detected by qPCR. The extracellular acidification rate (ECAR) of GCM-BMDM was detected by Seahorse bioenergy assay method. The protein levels of signal transducer and activator of transcription 6 (STAT6), signal transducer and activator of transcription 3(STAT3) and their phosphorylated forms p-STAT6 and p-STAT3 were examined by Western blot. Results GCM stimulated mRNA expression of Hif-1α and a variety of M1 and M2 oncogenic mediators in BMDM (P＜0.01). GCM upregulated the mRNA expression of multiple glycolysis associated genes (P＜0.01) and increased glycolysis level in BMDM (P＜0.001). The glycolysis level was reduced by incubation with lactate dehydrogenase inhibitor stiripentol (STP) (P＜0.05). STP down-regulated mRNA levels of Hif-1α and oncogenic mediators Il-1β, Il-6, Ido and Cd163 in GCM-BMDM(P＜0.01). STP decreased the ratio of p-STAT3 /STAT3 and p-STAT6 /STAT6 in GCM-BMDM (P＜0.05). Conclusions GCM increases glycolysis level and induces the pro-tumoral phenotypes in GAMs through promoting transcriptional regulation mediated by HIF-1α, STAT3 and STAT6. STP can effectively reduce glycolysis and transcriptional expression of oncogenic mediators in GAMs.

Value of blood urea nitrogen level in prediction of MACE after PCI in patients with acute coronary syndrome

GAO Xiaoqian, YU Huahui, LIU Sheng, JIAO Xiaolu, LYU Qianwen, ZHANG Ming, QIN Yanwen

2024, 44(5):  606-612.  doi:10.16352/j.issn.1001-6325.2024.05.0606

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Objective To explore the relationship and predictive function of blood urea nitrogen (BUN) level in association with major adverse cardiovascular event(MACE) subsequent to percutaneous coronary intervention(PCI) among patients diagnosed with acute coronary syndrome (ACS). Methods Between April 2017 and November 2017, ACS patients undergoing their initial PCI at the Department of Cardiology, Beijing Anzhen Hospital, Capital Medical University were recruited. Following a 36-month follow-up period, 487 patients were included as 114 subjects experiencing MACE and 373 subjects without MACE. The Cox proportional hazards regression model was utilized to assess the risk ratio of MACE and its 95% confidence interval (CI) concerning BUN levels and ACS post-PCI. Additionally, the area under the receiver operating characteristic (ROC) curve (AUC) was applied to assess the predictive effect of BUN. Results After adjusting for confounding variables, Cox proportional hazards regression analysis revealed that the risk of MACE was approximately four times higher in the high BUN level group compared to the low BUN level group (OR=4.722,95% CI 1.716-12.993). Upon inclusion of BUN in both the basic prediction model and SCORE model for cardiovascular event risk, there was a significant increase in the AUC area (P＜ 0.001). Conclusions BUN levels can independently predict the risk of MACE following PCI in ACS patients, irrespective of traditional risk factors.

Application of different methods used to transfect small molecule NADPH into cells

LI Haoyue, DU Wenjing, LI Wei

2024, 44(5):  613-618.  doi:10.16352/j.issn.1001-6325.2024.05.0613

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Objective To compare the three different methods for direct transfection of small exogenous nicotinamide adenine dinucleotide phosphate(NADPH) into cells by a non-metabolic pathway. Methods Three different transfection reagents (X-tremeGENE^TM HP DNA, Lipofectamine^TM RNAiMAX and Lipofectamine^TM 2000) were used to find the transfection efficiency of NADPH into human osteosarcoma cell line U2OS and mouse embryonic fibroblast cell line 3T3L1. Their effects on adipocyte differentiation were compared by Oil Red O staining. Results Transfection of NADPH with X-tremeGENE HP DNA transfection reagent effectively increased intracellular NADPH level(P＜0.001). With the increase of NADPH transfection concentration (10 μmol/L NADPH and 10 μL transfection reagent), the level of NADPH in cells increased in a dose-dependent manner. In addition, three transfection reagents were used to transfect NADPH in 3T3L1 preadipocytes. Only cells transfected with NADPH using X-tremeGENE HP DNA transfection reagent contained more lipids than control cells (P＜0.001). Conclusions Transfection of NADPH with X-tremeGENE HP DNA transfection reagent effectively increases the level of NADPH in cells.

RNA-binding protein RBPMS regulates proliferation and migration of acute myeloid leukemia cells line THP-1

ZHAO Lingyan, HE Liu, MA Yanni, YU Jia

2024, 44(5):  619-625.  doi:10.16352/j.issn.1001-6325.2024.05.0619

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Objective To investigate the impact of the RNA-binding protein with multiple splicing(RBPMS), which has multiple splicing functions, on the function of acute myeloid leukemia cells line THP-1 carrying the MLL-AF9 fusion gene. Methods The expression of RBPMS in the hematopoietic system was analyzed using the GEO database; RBPMS over-expression in THP-1 cells was achieved through lentiviral infection. The efficiency was detected by RT-qPCR and Western blot; The effect of RBPMS over-expression on the THP-1 cell transcriptome was evaluated by high-throughput sequencing. The impact of RBPMS over-expression on the proliferation and migration of leukemia cells was examined using CCK-8 assay and flow cytometry, respectively. Results RBPMS expressed in acute myeloid leukemia stem cells carrying the MLL-AF9 fusion gene was found to be inhibited. Over-expression of RBPMS significantly reduced the proliferation and migration of THP-1 cells. Conclusions RBPMS inhibits cell proliferation and migration of acute myeloid leukemia cells line carrying the MLL-AF9 fusion gene.

Na⁺/Ca²⁺ exchanger inhibitor SN-6 and verapamil reduce aldosterone synthase expression in adrenocortical carcinoma cell line NCI-H295R

WANG Yu, GAO Yinjie, REN Weidong, TONG Anli

2024, 44(5):  626-629.  doi:10.16352/j.issn.1001-6325.2024.05.0626

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Objective To investigate the effect of Na⁺/Ca²⁺ exchanger inhibitors SN-6 and calcium channel blocker verapamil on potassium-stimulated expression of aldosterone synthase in adrenocortical carcinoma cell line NCI-H295R(H295R). Methods H295R cells were treated in different groups, including control group, K⁺ (15 mmol/L) treated group, verapamil (10 μmol/L) group, SN-6 (10 μmol/L) group, combination of K⁺/verapamil treated group, combination of K⁺/SN-6 treated group, combination of K⁺/verapamil and SN-6 treated group. Real-time PCR was used to detect the mRNA expression of aldosterone synthetase (CYP11B2), and FLIPR Calcium 6 was used to detect the intracellular calcium ion level. Results Compared with the control group, K⁺ stimulated CYP11B2 mRNA expression. Both SN-6 and verapamil decreased K⁺ induced CYP11B2 mRNA expression(P＜0.01). Compared with the K⁺+SN-6 treated group, the K⁺,SN-6 and verapamil treated group all significantly inhibited CYP11B2 mRNA expression (P＜0.001). Both of SN-6 and verapamil significantly reduced the intracellular calcium level stimulated by K⁺ (P＜0.0001). Conclusions Both SN-6 and verapamil inhibit K⁺-induced expression of aldosterone synthase in adrenocortical carcinoma H295R cells. The inhibitory effect is more significant when treated by the combination of verapamil and SN6.

Bioinformatics analysis of cognitive dysfunction associated with diabetes mellitus with TREM2 mutation

LIU Xiao, WANG Zhaohui, WEI Xinyi, ZHOU Yue, ZHAO Li, WANG Yue, LI Junfa

2024, 44(5):  630-636.  doi:10.16352/j.issn.1001-6325.2024.05.0630

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Objective To explore the hub genes and the potential targets in the treatment of diabetes with TREM2 mutation-related cognitive dysfunction with bioinformatics analysis. Methods The cases of differential genes (DEGs) of diabetes mellitus and TREM2 mutation-related cognitive dysfunction were obtained respectively by microarray data analysis, the common differential genes were obtained by intersection between the two diseases. GO analysis, KEGG and Reactome pathway analysis were performed on the selected differential genes. The protein-protein interaction(PPI) network was constructed using online database. Finally, the effects of diabetes and TREM2 on spatial learning and memory of mice were detected by water maze, and the expression of hub gene SNAP25 was detected by Western blot. Results In both datasets, 19 genes showed similar changes, mainly enriched in biological processes and pathways related to neurons and metabolism. According to PPI analysis, DNER, GFAP, GRM5 and SNAP25 were identified as hub genes. Trem2 gene knockout aggravated spatial learning and memory impairment in diabetic mice. The expression of SNAP25 in hippocampus of diabetes mice was significantly increased, and then decreased after Trem2 knockout. Conclusions This study identified 19 TREM2-related genes in diabetes with cognitive dysfunction, among which 4 hub genes were found. These results provide a new experimental basis for the treatment of diabetes patients with cognitive impairment.

Regulation of TMEFF1 on proliferation of neuroblastoma cell lines

JIA Anna, GUO Jinxin, ZHANG Xuan, ZHAN Shijia, YU Yongbo, GUO Yongli, CHANG Yan

2024, 44(5):  637-644.  doi:10.16352/j.issn.1001-6325.2024.05.0637

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Objective To investigate the regulatory effects of transmembrane protein with EGF-like and two follistatin-like domain 1(TMEFF1/tomoregulin 1) on the proliferation and migration of neuroblastoma (NB) cell line SN-K-BE(2). Methods TMEFF1 mRNA in neuroblastoma cell lines SN-K-BE(2), IMR32, SK-N-SH, SK-N-AS and normal cell lines MCF10A and hTERT RPE-1 was detected by RT-qPCR. Small interfering RNA (siRNA) was used to construct transient knockdown TMEFF1 normal and SN-K-BE (2) NB cell lines. The effect of transient knockdown of TMEFF1 mRNA was examined by RT-qPCR. After transfection of SN-K-BE (2) cells with siRNA, cell proliferation was detected by colony-forming unit assay and real-time cell analysis (RTCA). The positive rate of Ki-67 cells was determined by immunofluorescence staining. CellTiter-Glo (CTG) was used to detect cell activity. Stable knockdown of TMEFF1 was performed in SK-N-BE(2) cells by short hairpin RNA (shRNA) lentiviral infection, the stable outcomes of TMEFF1 was also detected by RT-qPCR. Cell proliferation was detected by coloning formation, RTCA and CTG. In addition, the positive rate of Ki-67 cells was detected by immunofluorescence,the cell mobility was measured by cell scratch assay. Results Compared with normal cell lines, the expression of TMEFF1 in NB cell lines was significantly higher (P＜0.001). Transient or stable knockdown of TMEFF1 had no significant effect on the proliferation of normal cell lines. However, by knocking down TMEFF1 decreased cell proliferation of SN-K-BE (2) cells as showen RTCA and colony-forming unit assay (P＜0.05). Immunofluorescence results showed that the rate of Ki-67 positive cells was reduced (P＜0.001). CTG results showed that the cell activity was decreased(P＜0.01). Cell scratch assay also showed that the cell migration ratio in the knockdown group was significantly lower than that of control group (P＜0.01). Conclusions TMEFF1 is a protein specifically highly expressed in neuroblastoma. TMEFF1 knockdown has no effect on normal cell lines, but significantly inhibited the proliferation and migration of NB cells, suggesting that TMEFF1 may play an important role in the occurrence and development of neuroblastoma, and so may be a potential target for NB targeting therapy.

Melatonin inhibits H₂O₂-induced oxidative stress and apoptosis marker protein expression in human umbilical vein endothelial cells

YANG Hanyi, NING Jiayi, WANG Xiaolan, ZHANG Yimeng, XIE Tingke, CHEN Yixuan, HAN Jing

2024, 44(5):  645-650.  doi:10.16352/j.issn.1001-6325.2024.05.0645

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Objective To investigate the effects of melatonin (MT) on oxidative stress and expression of apoptosis marker protein in human umbilical vein endothelial cells (HUVECs) induced by hydrogen peroxide (H₂O₂). Methods HUVECs were divided into blank group and H₂O₂ (100, 200, 300, 400, 600, 700 mmol/L) groups.CCK8 method was used to select the best concentration for modeling. To clarify the effect of melatonin on oxidative stress injury caused by H₂O₂, HUVECs were divided into control group, H₂O₂ group and melatonin group. Western blot was used to detect the expression of p-p65/p65, SOD2 and cleaved-caspase 3. The activity of super-oxide dismutase (SOD), the concentration of malondialdehyde (MDA) and catalase (CAT) were measred by commercially available kits. Reactive oxygen species (ROS) staining was used to detect ROS content in HUVECs. Results H₂O₂ increased the expression of p-p65/p65 and cleaved-caspase 3(P＜0.001), decreased the expression of SOD2(P＜0.05)its biological activity(P＜0.001) and the concentration of CAT(P＜0.01), elevated the concentration of MDA(P＜0.001) and the level of ROS(P＜0.001)in HUVECs. Compared with H₂O₂ group, MT treatment decreased the expression of p-p65/p65(P＜0.001), SOD2(P＜0.001)and cleaved-caspase 3(P＜0.01), increased the expression of SOD2(P＜0.001)as well as biological activity of SOD(P＜0.001) and the concentration of CAT(P＜0.05) reduced the concentration of MDA(P＜0.001) and the level of ROS(P＜0.001)in HUVECs. Conclusions Melatonin plays a protective role in oxidative stress injury of HUVECs induced by H₂O₂ through up-regulating the expression of SOD2 and down-regulating the expression of p-p65/p65 and cleaved-caspase 3.

Protein expression and subcellular distribution of centriolar component SAS-6 in mouse oocyte meiosis

SONG Ke, SONG Ke, LI Jingyu, MA Wei, YANG Xiaokui

2024, 44(5):  651-657.  doi:10.16352/j.issn.1001-6325.2024.05.0651

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Objective To study the expression and subcellular distribution pattern of centriolar protein SAS-6 during meiosis of mouse oocytes. Methods Immunefluorescence was conducted to analyze the subcellular distribution pattern of SAS-6 in Chinese hamster ovary cell line(CHO cell) during mitosis. Western blot was applied to detect the protein expression of SAS-6 in mouse oocyte meiosis. Immunofluorescence microscopy carried out to determine the subcellular distribution of SAS-6 and its association with microtubule organizing centers (MTOCs) and in vesicles during the oocyte meiosis. Results SAS-6 was colocalized with Pericentrin on spindle poles during the mitotic process in somatic cells. In mouse oocytes, SAS-6 was stably and consistently expressed at all stages of meiosis and specifically aggregated on GM130-positive vesicles, but not in MTOCs. The results suggested that centriolar protein SAS-6 was still expressed in oocytes lacking centrosomes, and localized in GM130-positive vesicles but expressed on percientrin-positive MTOCs. Conclusions SAS-6 may play a role in spindle cortical migration through regulating vesicles during oocyte meiosis.

Correlation between circulating CD4⁺CD45RA⁺CD62L⁺ T cells and prognosis of metastatic non-small cell lung cancer treated with EGFR-TKI

CAO Chenxin, TANG Hui, GENG Ruixuan, GUO Fuping, BAI Chunmei, WANG Yingyi, LI Taisheng

2024, 44(5):  658-664.  doi:10.16352/j.issn.1001-6325.2024.05.0658

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Objective To explore the correlation between circulating lymphocyte profiles and the outcomes of non-small cell lung cancer (NSCLC) patients undergoing epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) therapy. Methods A retrospective cohort of 40 patients who received EGFR-TKI therapy at Peking Union Medical College Hospital was designed. Circulating lymphocyte subsets were collected using flow cytometry for dynamic monitoring during EGFR-TKI therapy. The survival of each patient was followed up by telephone call. The correlation of baseline and dynamic change of the peripheral blood circulating lymphocyte subsets and survival were further analyzed. Results Patients who responded to EGFR-TKI therapy had significantly higher baseline circulating CD4⁺CD45RA⁺CD62L⁺ T-cell counts during dynamic monitoring. The median progression-free survival (PFS) for the entire population was 27.1 months; however, overall survival (OS) was not achieved. The median PFS did not differ significantly between patients with higher and lower baseline CD4⁺CD45RA⁺CD62L⁺ T-cell counts. Furthermore, dynamic changes in CD4⁺CD45RA⁺CD62L⁺ T-cell counts were significantly correlated with not only the response to EGFR-TKI therapy response, but also PFS. PFS of patients with stable or increased CD4⁺CD45RA⁺CD62L⁺ T-cell counts during the EGFR-TKI therapy was significantly longer than that of patients with decreased CD4⁺CD45RA⁺CD62L⁺ T-cell counts (29.1 months vs. 9.4 months; P＜0.001). Conclusions Higher baseline circulating CD4⁺CD45RA⁺CD62L⁺ T-cell counts indicate a better EGFR-TKI response, and dynamic changes in CD4⁺CD45RA⁺CD62L⁺ T-cell counts are associated with prolonged PFS.

miR-135a-5p alleviates morphine tolerance by targeting the regulation of CXCL12 expression

HE Huamei, LU Wei

2024, 44(5):  665-672.  doi:10.16352/j.issn.1001-6325.2024.05.0665

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Objective Investigating the effect of miR-135a-5p in regulating of CXCL12 on the formation of morphine tolerance in C57BL/6J mice. Methods Totally 64 mice were randomly divided into the following groups: control (NS) group, morphine tolerance (MT) group, CXCL12 inhibitor (CXCL12-siRNA) group, inhibitor negative control (NC-siRNA) group, miR-135a-5p agonist (miR-agomir) group, agonist negative control (miR-NC) group, miR-135a-5p agonist+CXCL12 overexpression (miR-agomir+LV-CXCL12) group, and miR-135a-5p agonist+CXCL12 over-expression negative control (miR-agomir+LV-control) group. Morphine tolerance models were established by subcutaneous injection. The tail-flick test was used to measure the thermal tail-flick latency (TL) before and after drug administration in each group. After modeling, the mice were euthanized under anesthesia, and L4~L5 spinal cord tissues were collected. RT-qPCR method was used to detect the expression of miR-135a-5p and CXCL12 mRNA, Western blot was used to detect the expression of CXCL12 proteins, and a dual luciferase reporter gene assay was used to detect the targeting relationship between miR-135a-5p and CXCL12. Results 1)The morphine-tolerant mouse model demonstrated a significant increase in CXCL12 mRNA and protein expression(P＜0.05) and a significant downregulation of miR-135a-5p (P＜0.05) compared to the NS group. 2)Silencing Cxcl12 and over-expression of miR-135a-5p both increased the thermal pain threshold in morphine-tolerant mice, significantly slowed down the occurrence and progression of morphine tolerance. 3)Compared to the miR-NC group, the miR-agomir group showed a decrease in both CXCL12 mRNA and protein expression(P＜0.05). 4)Dual-luciferase reporter gene experiments demonstrated that miR-135a-5p was targeted at Cxcl12. 5)Over-expression of CXCL12 reversed the thermal pain-protective effect of miR-135a-5p in morphine-tolerant mice. Conclusions It is demonstrated by the results above that miR-135a-5p alleviates the formation of morphine tolerance by suppressing the expression of CXCL12.

Allicin inhibits the proliferation of oral cancer cells line CAL27

WANG Tingting, XIAO Ning, LIN Dianxin, DONG Haitao

2024, 44(5):  673-676.  doi:10.16352/j.issn.1001-6325.2024.05.0673

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Objective To explore the effect of allicin on the proliferation of oral tongue squamous cell carcinoma(OTSCC) cells line(CAL27). Methods Human CAL27 cells were used in this study. The cells were incubated with different concentrations of allicin (5, 10, 20, 40, 60, 100, 200, 400 μmol/L) and control DMSO. CCK8 assay was performed to detect cell proliferation. RT-qPCR method was used to detect the expression of proliferation related marker ki-67; Transcriptome sequencing and pathway enrichment analysis were performed. Results Lower concentrations (20 μmol/L) of allicin inhibited CAL27 cells proliferation (P＜0.01).Allicin significantly inhibited the expression of ki-67 in CAL27 cells at lower concentrations(20 μmol/L, P＜0.01). Allicin inhibited tumor related microRNA expression, especially miR-200a and miR-199a1. Conclusions Lower concentration of allicin may significantly inhibit the proliferation of oral cancer cells by inhibition of tumor related microRNAs as a potential mechanism.

Identification of the pathogenic variants in two Chinese patients with primary ciliary dyskinesia

ZHENG Haixia, ZHOU Wangji, TIAN Xinlun, LIU Yaping

2024, 44(5):  677-682.  doi:10.16352/j.issn.1001-6325.2024.05.0677

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Objective To characterize the clinical features and to identify the pathogenic variants in two Chinese patients with primary ciliary dyskinesia (PCD). Methods The clinical data and peripheral blood sample from the patients were collected, and genomic DNA was subsequently extracted from the peripheral blood. Candidate pathogenic variants were identified using whole exome sequencing (WES) and further confirmed by Sanger sequencing technology. Finally, the pathogenicity of the variants was predicted through bioinformatic analysis. Results Two Chinese patients with PCD had diffuse bronchiectasis cpmlicated with recurrent infection and the decreased level of nasal nitric oxide (nNO). WES results showed that both patients carried frameshift mutations in known pathogenic genes of PCD. Patient 1 carried a homozygous variant in outer dynein arm docking complex subunit 1 (ODAD1) (NM_144577): c.702_705dupGCAG (p.P236Afs*11) and patient 2 carried a hemizygous variant in dynein axonemal assembly factor 6 (DNAAF6) (NM_173494): c.532_533delCT (p.L178Sfs*2). Neither variant had been recorded in The Human Gene Mutation Database (HGMD). Both frameshift variants caused changes in the open reading frame, which resulted in premature termination codon. According to the American College of Medical Genetics and Genomics (ACMG) guidelines, c.702_705dupGCAG in ODAD1 was categorized as a pathogenic variant (PVS1+PM2+PM3+PP4) and c.532_533delCT in DNAAF6 was categorized as a pathogenic variant (PVS1+PM2+PM3+PP4). Conclusions The novel variants c.702_705dupGCAG found in ODAD1 and c.532_533delCT found in DNAAF6 are pathogenic and support their PCD diagnosis for the two patients, respectively. These results may enrich the mutation spectrum of the ODAD1 and DNAAF6.

BM-MSCs delay the senescence of naive CD8⁺T cells

GAO Jingxi, ZHAO Xiaoyan, ZHU Xingyu, SUN Zhao, HAN Qin, ZHAO Chunhua

2024, 44(5):  683-689.  doi:10.16352/j.issn.1001-6325.2024.05.0683

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Objective To verify the effect of bone marrow mesenchymal stem cells (BM-MSCs) in alleviating immune senescence, and to explore the main immune cell population improved by BM-MSCs. Methods Mouse spleen lymphocytes were isolated and stimulated to proliferate for 7 days for constructing a replicative aging model. Flow cytometry was used to detect the p16ink4a(p16) and p21cip1(p21) expression by T cell subpopulation in the young control group, the replicative senescence control group and the BM-MSCs co-cultured group. Results In the replicative senescence model of T lymphocytes, it was observed that CD8⁺T cells senescent significantly as compared with CD4⁺T cells after continuous proliferation. Among the naive cells and effector cell subsets of CD8⁺T cells, effector cell senescence was the most significant. BM-MSCs co-culture had no significant effect on senescent effector cells, and mainly alleviated the senescence of CD8⁺T cells by delaying the senescence of naive T cells(P＜0.01,P＜0.001). Conclusions BM-MSCs co-culture can alleviate the replicative senescence phenotype of T cells and has a more significant anti-senescence effect on CD8⁺T cells by inhibiting the initial senescence of T cells as a major mechcanism.

Quantitative analysis of trace-level proteins for assessing inflammation through clinical sampling

YE Yuxin, LI Huixian, YAN Tao, ZHANG Yun, SUN Yulin

2024, 44(5):  690-698.  doi:10.16352/j.issn.1001-6325.2024.05.0690

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Objective To compare the effectiveness between qPCR and trace-level proteins quantitative analysis in detecting the expression of inflammation-associated factors caspase-1 and IL-1β in peripheral blood monocytes from breast cancer patients. Methods Pre- and post-operation peripheral blood samples were collected from 10 breast cancer patients. Monocytes were subsequently isolated and activated by incubation with lipopolysaccharide. The mRNA and protein expression level of caspase-1 and IL-1β in monocytes were examined using qPCR and trace-level proteins quantitative analysis, respectively, and the data were normalized by β-actin. The differences between the two detection methods were compared and evaluated for strengthens and weakness. Results The vast majority of patients showed a notable increase in the number of peripheral blood monocytes after surgical operation. At the mRNA level, compared to preoperative levels, postoperative caspase-1 and IL-1β were both increased in 50% (5/10) and 40% (4/10) of patients, respectively, while the rest showed either a decrease (40% and 50%, respectively) or no significant change (both 10%). In contrast, the results of trace-level proteins quantitative analysis revealed that 60% (6/10) of patients exhibited increased level of postoperative caspase-1 and IL-1β proteins, while the remaining showed a decrease (30% and 20%, respectively) or no significant change (10% and 20%, respectively). Additionally, 50% of patients displayed a significant increase in caspase-1 and IL-1β active cleavage bands by electrophoresis postoperatively. Notably, inconsistent changes in caspase-1 and IL-1β mRNA and protein level after surgery were observed in 30% and 40% of patients respectively. Conclusions The trace-level proteins quantitative analysis is recommended due to its advantages including minimal sample amount requirement, high sensitivity, good repeatability and relatively simple operative procedure. Therefore, it is particularly suitable for assessing the inflammatory status of of patients through clinical sampling of peripheral blood.

Pathological changes and metabolome analysis of liver in obese mouse models

DING Baofeng, WANG Xiaoshuang, WANG Fang, YU Jia

2024, 44(5):  699-704.  doi:10.16352/j.issn.1001-6325.2024.05.0699

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Objective To detect the pathological changes in obese mice liver and explore the effects of obesity on hepatic metabolism. Methods Obese mouse model was successfully constructed by consecutive 24 weeks of high fat diet (HFD), while control group was fed with a standard chow diet (CD). After 24 weeks, mice liver was dissected, and the pathological characteristics of liver were observed with hematoxylin-eosin (HE) staining and Sirius red staining. The metabolome of liver was detected by ultra-high performance liquid chromatography-mass spectrometry(UPLC-MS) after extraction by 80% methanol. Principal component analysis (PCA), differential metabolite analysis and KEGG pathway enrichment analysis were performed to find the metabolic changes in the liver of obese mice. Results After 24 weeks of high fat diet, the body weight and liver weight of HFD group mice were much higher than that of CD group mice. Besides, HFD group mice liver showed severe steatosis and slight fibrosis. There were also significant differences in liver metabolites between HFD and CD groups, and the metabolic changes were mainly enriched in pathways of phenylalanine metabolism, citric acid cycle (TCA cycle), sphingolipid metabolism, arginine biosynthesis and primary bile acid biosynthesis. Conclusions The obese mouse model was successfully constructed, and the pathological characteristics of obese mice liver were elucidated. At the same time,the main metabolic pathway changes of obese mice liver were further revealed by metabonomics analysis.

Clinical Sciences

Assessment models of dental fear and anxiety in children based on psysiological data

ZHANG Peng, ZHANG Yuyang, WANG Fei, LI Xiying

2024, 44(5):  705-708.  doi:10.16352/j.issn.1001-6325.2024.05.0705

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Objective To explore the neurophysiological responses of patients′ fear in real treatment scenarios, and to construct a assessment model of children′s dental fear and anxiety (DFA) based on physiological data. Methods Totally 166 children under the age of 14 who received treatment at the Beijing Children′s Hospital Department of Stomatology were included in the study. The Chinese version of the modified Children′s Fear Survey Schedule-Dental Subscale (CFSS-DS) was used to assess dental fear and anxiety scores in children. A wrist-worn physiological data collection device was used to continuously measure subjects′ heart rate (HR) and galvanic skin response (GSR) during the treatment. Repeated measures analysis of variance was used to test for differences in physiological data before, during and after treatment. Pearson′s correlation analysis was used to analyze the relationship between physiological data and CFSS-DS scores before, during, and after treatment. A Least Absolute Shrinkage and Selection Operator(LASSO) regression model was established to assess dental fear and anxiety among children, and Leave-One-Out (LOO) was used for cross-validation. Results There were significant differences in the changes of each dimension of physiological data before, during, and post treatment(P＜0.05). The Pearson′s correlation coefficient (r) between the LASSO model predictive values and the CFSS-DS scores was 0.30 (P＜0.05), which indicated a good predictive accuracy of the model. Conclusions The assessment model might achieve objective and automated measurement of dental fear and anxiety in children.

Effect of age on the success rate of lacrimal probing in the congenital nasolacrimal duct obstruction

WU Xingzhi, SUN Hua, DING Jingwen

2024, 44(5):  709-713.  doi:10.16352/j.issn.1001-6325.2024.05.0709

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Objective To explore the effect of age factor on the success rate of lacrimal probing in the congenital nasolacrimal duct obstruction (CNLDO). Methods This report was based on a single-center retrospective case series study. The gradient therapeutic regimen was adopted, and lacrimal probing was undertaken to treat CNLDO when conservative treatment was ineffective. The patients were categorized into 4 groups by age: 0-＜6 months,6-＜12 months,12-＜24 months and 24-60 months. The success rate of probing in each group was compared and the correlation with age was analyzed. Results A total of 314 eyes of 269 patients were included. The success rates of primary probing in 4 groups were 86.8%, 91.7%, 73.6% and 37.8%, respectively. The overall success rate of probing in all patients was 84.7%(266/314). The decreased success rate of probing at age beyond 12 months was significantly correlated with increasing age(P＜0.001). No significant correlation was found with sex or laterality. Conclusions Staged treatment strategy is safe and effective. The success rate of probing is negatively correlated with age. Initial probing is recommended in children under 2 with failure of the conservative management.

Effect of oral probiotics on vaginal microbiota in patients with vaginitis during pregnancy

SONG Min, ZHANG Yuping, ZHU Ping, YANG Qiuhong, ZHANG Suxia, LIU Cuiying, LIU Yanping

2024, 44(5):  714-718.  doi:10.16352/j.issn.1001-6325.2024.05.0714

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Objective To evaluate the effect of oral probiotics containing 4 strains of lactobacillus on the vaginal microbiota and adjuvant treatment effect of patients with vaginitis during pregnancy. Methods A total of 395 cases were included, 171 cases in the control group received 7 days of antibiotic treatment, and 224 cases in the combined treatment group. After the combined use of antibiotics and probiotics for 7 days, they continued to take 1 probiotic pill daily for the 28th day. The patients received a vaginal microecological examination and clinical evaluation at study entry and 28 days to compare the vaginal lactobacillus between the two groups. Results There was a statistical difference in the proportion of vaginal lactobacillus and the efficacy of treatment between the two groups of patients (P＜0.05). The effective rates in the combined treatment group and the control group were 90.2% and 86.0% respectively, and the cure rates were 30.4% and 19.3% respectively. Conclusions Oral probiotics containing lactobacillus strains can increase the proportion of vaginal lactobacillus in patients with vaginitis during pregnancy which significantly facilitated the clinical treatment of vaginitis during pregnancy.

Mini Reviews

Research progress in the treatment of severe aplastic anemia

FU Chen, LI Ping

2024, 44(5):  719-723.  doi:10.16352/j.issn.1001-6325.2024.05.0719

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Severe aplastic anemia(SAA)is a critically ill disease of the blood system that progresses rapidly, has a high mortality rate, and a very poor prognosis. The application of drugs like androgens and Eltrombopag has significantly increased the survival rate of those with aplastic anemia (AA), compared with horse antithymocyte globulin(H-ATG) and cyclosporine, which was once the standard treatment. Patients who do not respond well to immunotherapy or experience a recurrence of symptoms post-treatment have the option to undergo hematopoietic stem cell transplantation (HSCT). The advancement of pre-treatment techniques for HSCT and the utilization of diverse medications following the transplantation procedure have contributed to reduced incidence of graft versus-host disease (GVHD) and improved transplant success. Allogeneic hematopoietic stem cell transplantation has become a hot topic in clinical research. This study summarizes the transplant and non transplant treatments of SAA in recent years, and analyzes its efficacy and related advantages and disadvantages.

Research progress of phosphodiesterase 5 in heart failure

YANG Yang, ZHANG Yarong, YANG Hongqin, WANG Jing, ZHAO Hongmei

2024, 44(5):  724-728.  doi:10.16352/j.issn.1001-6325.2024.05.0724

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Phosphodiesterase 5 (PDE5), a cyclic guanosine monophosphate (cGMP)-specific hydrolase, is targets cGMP produced by soluble guanosine cyclase (sGC) that is activated by nitric oxide (NO). It catalyzes the hydrolysis of the phosphodiester bond in cGMP and converts cGMP to the inactive 5′-GMP form. The cGMP-PKG axis dysfunction is one of the main causes of heart failure (HF) and causes cardiac remodeling. However, the clinical efficacy of PDE5 inhibitors in the treatment of heart failure is controversial. This paper summarizes the mechanism and research progress of PDE5 in heart failure in recent years, which has important significance for the clinical application of PDE5 in heart failure.

Progress in drug therapy for hypothalamic obesity

ZHANG Liyuan, DU Hanze, PAN Hui

2024, 44(5):  729-732.  doi:10.16352/j.issn.1001-6325.2024.05.0729

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Hypothalamic obesity (HO) is a complicated and uncommon disease affecting multiple regulatory pathways of energy intake and metabolism in the brain, regulation of the autonomic nervous system and peripheral hormonal signaling. Currently, drug treatment mainly involves stimulating sympathetic nerve excitation and fat tissue thermogenesis, thereby inhibiting rapid weight gain by increasing fat breakdown and energy consumption. In addition, some drugs can also act on the reward center of the brain to inhibit excessive food intake in HO patients. This article systematically introduces the clinical benefit data, adverse reactions, and application prospects of newly developed drugs such as oxytocin (OXT), phentermine and topiramate(Ph/T), tesofensine, setmelanotide, etc.

Medical Education

Investigation and analysis of the current situation of research capacity of professional graduates majoring in neurosurgery

LIU Zhenlei, JIAN Fengzeng, WU Hao

2024, 44(5):  733-737.  doi:10.16352/j.issn.1001-6325.2024.05.0733

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Objective In order to provide reference and basis for the scientific research capacity training of medical graduates, the current situation training for professional graduates majoring in neurosurgery was investigated and analyzed. Methods A questionnaire survey was conducted among graduates majoring in neurosurgery at Xuanwu Hospital. Possible relevant factors were analyzed, including scientific research awareness, scientific research initiative, scientific research interest, weekly science citation index^TM(SCI) literature reading, number of thesis published in Chinese core journals, number of thesis published in SCI journals, and participation in clinical and basic research project, number of registered clinical studies, experience in writing fund applications, supervisor′s scientific research requirements, participation in Journal Club activities. The current status of scientific research capacity building of graduates in our department and the factors of scientific research productivity were analyzed. Results Professional graduate majoring in neurosurgery had the highest weekly scientific research literature reading score (4.32±1.04), higher scientific research awareness, scientific research initiative, and scientific research interest scores (3.82-3.89).However, participation in scientific research (2.80±1.18) and thesis publication(2.50±1.52) were relatively low. Comparing graduates of different grades, scientific research awareness and initiative increase with raising grade. Most of first-year master candidates had the lowest scores in sum of relevant factors. In addition, the result of linear regression analysis suggested that scientific research interest and participation in scientific research had a significant impact on scientific research productivity(P＜0.05). Conclusions It is necessary to stimulate graduates′ interest in scientific research and encourage them to join research project and strengthen the capacity of setting research goals. Also more efforts are needed to optimize research and training environment in terms of tutors and institutions, thus to promote the building of research capacity of graduates to meet the comprehensive requirements for cultivation of medical talents.

Construction of a tiered training model for laparoscopic techniques in urology

DONG Dexin, ZHANG Yushi

2024, 44(5):  738-740.  doi:10.16352/j.issn.1001-6325.2024.05.0738

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With the development of laparoscopic technology in urology, the training of laparoscopic technology operator plays an extremely important role in the specialized education process from medical students to specialist physicians. With various medical students being included in standardized training for specialized physicians, many problems such as uneven quality of trainees appeared, lack of standardization in the training process, deviation in training target positioning, and insufficient quality control management methods. This article explored the construction of a standardized tiered training system for laparoscopic techniques in the urology department of Peking Union Medical College Hospital, including cultural construction, training mode system, assessment mechanism and trainers′ development. By constructing the special model for laparoscopic technology training in urology, it would lay a solid foundation and provide experience for building a standardized training system of laparoscopy in urology.