Study on the Role and Mechanism of METTL3 Mediating the Up-regulation of m6A Modified Long Non-coding RNA THAP7-AS1 in Promoting the Occurrence of Lung Cancer / 中国肺癌杂志

BACKGROUND@#Lung cancer is a major threat to human health. The molecular mechanisms related to the occurrence and development of lung cancer are complex and poorly known. Exploring molecular markers related to the development of lung cancer is helpful to improve the effect of early diagnosis and treatment. Long non-coding RNA (lncRNA) THAP7-AS1 is known to be highly expressed in gastric cancer, but has been less studied in other cancers. The aim of the study is to explore the role and mechanism of methyltransferase-like 3 (METTL3) mediated up-regulation of N6-methyladenosine (m6A) modified lncRNA THAP7-AS1 expression in promoting the development of lung cancer.@*METHODS@#Samples of 120 lung cancer and corresponding paracancerous tissues were collected. LncRNA microarrays were used to analyze differentially expressed lncRNAs. THAP7-AS1 levels were detected in lung cancer, adjacent normal tissues and lung cancer cell lines by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The diagnostic value of THAP7-AS1 in lung cancer and the relationship between THAP7-AS1 expression and survival rate and clinicopathological parameters were analyzed. Bioinformatics analysis, methylated RNA immunoprecipitation (meRIP), RNA pull-down and RNA-immunoprecipitation (RIP) assay were used to investigate the molecular regulation mechanism of THAP7-AS1. Cell proliferation, migration, invasion and tumorigenesis of SPC-A-1 and NCI-H1299 cells were determined by MTS, colony-formation, scratch, Transwell and xenotransplantation in vivo, respectively. Expression levels of phosphoinositide 3-kinase/protein kenase B (PI3K/AKT) signal pathway related protein were detected by Western blot.@*RESULTS@#Expression levels of THAP7-AS1 were higher in lung cancer tissues and cell lines (P<0.05). THAP7-AS1 has certain diagnostic value in lung cancer [area under the curve (AUC)=0.737], and its expression associated with overall survival rate, tumor size, tumor-node-metastasis (TNM) stage and lymph node metastasis (P<0.05). METTL3-mediated m6A modification enhanced THAP7-AS1 expression. The cell proliferation, migration, invasion and the volume and mass of transplanted tumor were all higher in the THAP7-AS1 group compared with the NC group and sh-NC group of SPC-A-1 and NCI-H1299 cells, while the cell proliferation, migration and invasion were lower in the sh-THAP7-AS1 group (P<0.05). THAP7-AS1 binds specifically to Cullin 4B (CUL4B). The cell proliferation, migration, invasion, and expression levels of phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), phosphoinositide-3 kinase, catalytic subunit delta (PIK3CD), phospho-phosphatidylinositol 3-kinase (p-PI3K), phospho-protein kinase B (p-AKT) and phospho-mammalian target of rapamycin (p-mTOR) were higher in the THAP7-AS1 group compared with the Vector group of SPC-A-1 and NCI-H1299 cells (P<0.05).@*CONCLUSIONS@#LncRNA THAP7-AS1 is stably expressed through m6A modification mediated by METTL3, and combines with CUL4B to activate PI3K/AKT signal pathway, which promotes the occurrence and development of lung cancer.

Traditional Chinese Medicine Regulating Mitochondrial Function in Prevention and Treatment of Liver Failure：A Review / 中国实验方剂学杂志

Liver failure （LF）， as a clinical syndrome of severe hepatocyte damage and liver dysfunction， has become a major obstacle to human health due to the triple superposition of high mortality， high morbidity， and high medical resource depletion. It is of great significance to further study the core factors of the disease and supplementary treatment methods to improve the survival rate of patients with LF. The pathogenesis of LF is complex， and mitochondrion is one of the sensitive organelles in hepatocytes and the central link of intracellular energy metabolism. A large number of studies have shown that the structure and function of mitochondria in hepatocytes are changed in LF， and the abnormal structure and function of mitochondria play an important role in the process of LF disease. Among them， multiple factors such as mitochondrial respiratory chain disorder， mitochondrial DNA damage， mitochondrial permeability transition pore opening， mitochondrial quality control imbalance， and mitochondrial oxidative stress are intertwined， forming a complex and unified whole network， which becomes the key node affecting the progression of LF. In recent years， researchers have begun to study drugs that can regulate the function of liver mitochondria to prevent and treat LF. With the deepening of research， traditional Chinese medicine has made breakthroughs in the prevention and treatment of LF. Many studies have confirmed that traditional Chinese medicine can play a role in the prevention and treatment of LF by protecting mitochondrial function， which can be summarized as reducing liver cell damage， inhibiting liver cell death， and promoting liver cell regeneration， so as to effectively compensate for liver function and promote the recovery of liver parenchyma quality and function. This article summarized the structure and function of mitochondria， the relationship between LF and mitochondria， and the research on the intervention of mitochondrial function in the field of traditional Chinese medicine to prevent and treat LF， so as to provide certain ideas and references for the clinical treatment of LF with traditional Chinese medicine.

Study on the antitussive and expectorant activities and mechanism of platycodin D based on metabolomics method / 药学学报

In this paper, the antitussive and expectorant activity of platycodin D (PD) were studied by constructing a mouse cough induced by concentrated ammonia water and a mouse trachea phenol red excretion model. The mechanism of antitussive and expectorant effect of PD was studied by metabolomics. The animal experiment was approved by the Animal Ethics Committee of Jiangxi University of Chinese Medicine (approval number: JZLLSC-20220739). Then mice were randomly divided into the normal, model, positive drug, PD low-dose, PD medium-dose and PD high-dose group. The antitussive and expectorant effects of PD were evaluated using a cough mouse model induced by concentrated ammonia water and a mouse tracheal phenol red excretion model, respectively. UHPLC-LTQ-Orbitrap-MS was used to identify the metabolites of mouse lung tissue, and multivariate statistical analysis method of orthogonal partial least squares discriminant analysis (OPLS-DA) was used for metabolites profile analysis. The differential metabolites were screened by variable projected importance value (VIP) and t-test results. Pathways for enrichment of differentiated metabolites were analyzed using the MetaboAnalyst platform. The comparative method was applied to analyze the differences in mechanisms of PD, Deapio-platycodin D (DPD) and total platycosides fraction. The results showed that PD at different concentrations could significantly prolong (P < 0.05) the incubation period of cough mice induced by ammonia water, reduce the coughs frequency, and significantly increase (P < 0.05) the amount of phenol red excretion in phenol red excretion model mice. PD could regulate 6 metabolic pathways of phenylalanine, tyrosine and tryptophan biosynthesis, linoleic acid metabolism, phenylalanine metabolism, glycerophospholipid metabolism, and tyrosine metabolism to exert antitussive effect. It could also regulate 8 metabolic pathways of linoleic acid metabolism, glyoxylic acid and dicarboxylic acid metabolism, glycerol phospholipid metabolism, citric acid cycle and arachidonic acid metabolism to exert an expectorant effect. However, only linoleic acid metabolism and glycerophospholipid metabolism could be regulated by the PD, total platycosides fraction and DPD, which may be ascribed to the structural difference of the platycosides and the interaction between platycosides and the intestinal microbiota. Functional analysis showed that these metabolic pathways are closely related to the regulatory mechanisms of anti-inflammatory response, immune function regulation, neurotransmitter release, cell signal transduction, energy metabolism and cell apoptosis. This study shows that PD possesses good antitussive and expectorant activities. In addition, the mechanism difference of PD, total platycosides fraction and DPD imply that the apiose in PD and the interaction between PD and intestinal microbiota could exert an important effect on the antitussive and expectorant mechanism of the platycosides.

Retrospective analysis on the effect of prevention and treatment of senile pulmonary tuberculosis in Hubei Province / 公共卫生与预防医学

Objective To retrospectively analyze the prevention and control effect and epidemic characteristics of elderly tuberculosis in Hubei Province from 2016 to 2020, and to provide a scientific basis for the prevention and treatment of elderly tuberculosis in Hubei Province． Methods The data on tuberculosis patients aged 60 and above who registered their current address in Hubei Province from 2016 to 2020 were collected and analyzed. The registration rates and composition ratios were analyzed using χ2 test and χ2 test for trend. Results A total of 135 976 tuberculosis patients were reported in Hubei from 2016 to 2020. The annual average registration rate of elderly tuberculosis among the elderly registered residence population (referring to the registration rate of elderly registered residence population aged 60 and above as the denominator, and tuberculosis patients aged 60 and above as the numerator) was 263.51/100 000. The highest rate was 300.02/100,000 in 2017, and the lowest was 188.19/100,000 in 2020 (χ2=70,227.603, P2trend=40.448，P2trend=740.911, P2trend=380.557, P2trend=323.764, P<0.001). Conclusion The elderly population with pulmonary tuberculosis in Hubei Province shows a downward trend. It is necessary to focus on the efforts of designated hospitals to proactively identify cases, increase the proportion of confirmed cases, maintain a high tracking in place, reduce medical delays, and ensure the effectiveness of tuberculosis prevention and treatment for the elderly.

Research progress on drug treatment and drug resistance mechanism of gastrointestinal stromal tumors / 中国临床药理学与治疗学

Gastrointestinal stromal tumors (GIST) are the most common mesenchymal-derived tumors of the gastrointestinal tract. Tyrosine kinase inhibitors (TKIs) are the cornerstone of GIST therapy, but mutations in resistance genes pose many problems for treatment, especially the heterogeneity of KIT resistance mutations. In recent years, with the release of a number of GIST related drug research and experimental results, the great potential of targeted therapy, immunotherapy and combination therapy to treat GIST in different directions has been revealed, providing more therapeutic directions for GIST. This article will review the experimental research and future direction in recent years.

Effect of safflower yellow on learning and memory ability of APP/PS1 at different months based on TLR4/NF-KB signaling pathway / 中国药理学通报

Aim To investigate the effect of safflower yellow (SY) on learning and memory ability of APP/ PS1 mice at different disease stages, and to explore the mechanism of SY anti- Alzheimer's disease by using 3-,6- and 9-month-old APP/PS 1 transgenic mice as experimental animal models. Methods Behavioral experiments were conducted to observe the effects of SY on learning and memory of APP/PS1 mice of different months. ELISA was used to detect the effect of SY on the expression of inflammatory factors in cortex of mice of different months. Western blot was used to detect the microglia activation marker protein, and its mechanism of action was further analyzed. Results SY could enhance the learning and memory ability of mice aged 3, 6 and 9 months, reduce the content of IL-6 and increase the content of TGF-β1 in brain tissue, up-regulate the expression levels of arginase-1 (arg-1) and triggering receptor expressed on myeloid cells 2 (tREM2) in brain tissue of mice of different months, and down-regulate the expression levels of inducible nitric oxide synthase (iNOS), Toll-like receptors 4 (tlr4) and nuclear factor-kappa B (nf-KB). Conclusions Compared with 3- and 9-month-old mice, SY is the most effective in improving learning memory in 6-month-old APP/PS1 mice. SY inhibits TLR4/NF-KB pathway activation by inducing TREM2 expression in brain tissue of APP/PS 1 transgenic mice, promotes microglia phenotype shift to anti-inflammatory phenotype, reduces chronic neuroinflammatory response, and improves learning memory in APP/PS1 mice at all months of age.

Expert Consensus on Clinical Diseases Responding Specifically to Traditional Chinese Medicine：Aural Vertigo / 中国实验方剂学杂志

Aural vertigo frequently encountered in the otolaryngology department of traditional Chinese medicine （TCM） mainly involves peripheral vestibular diseases of Western medicine， such as Meniere's disease， benign paroxysmal positional vertigo， vestibular neuritis， and vestibular migraine， being a hot research topic in both TCM and Western medicine. Western medical therapies alone have unsatisfactory effects on recurrent aural vertigo， aural vertigo affecting the quality of life， aural vertigo not relieved after surgery， aural vertigo with complex causes， and children's aural vertigo. The literature records and clinical practice have proven that TCM demonstrates unique advantages in the treatment of aural vertigo. The China Association of Chinese medicine sponsored the "17th youth salon on the diseases responding specifically to TCM： Aural vertigo" and invited vertigo experts of TCM and Western medicine to discuss the difficulties and advantages of TCM diagnosis and treatment of aural vertigo. The experts deeply discussed the achievements and contributions of TCM and Western medicine in the diagnosis and treatment of aural vertigo， the control and mitigation of the symptoms， and the solutions to disease recurrence. The discussion clarified the positioning and advantages of TCM treatment and provided guidance for clinical and basic research on aural vertigo.

Improvement effect of Phellodendron amurense polysaccharides on gouty nephropathy in rats and its mechanism / 中国药房

OBJECTIVE To study the effects of Phellodendron amurense polysaccharides （PAP） on improving gouty nephropathy （GN） in rats， and to investigate its mechanism primarily by interfering the p38 mitogen-activated protein kinase （p38 MAPK）/nuclear factor-κB（NF-κB）/tumor necrosis factor-α（TNF-α）. METHODS Sixty rats were randomly divided into normal group （water）， model group （water）， allopurinol group （positive control， 20 mg/kg）， PAP high-dose， medium-dose and low-dose groups （100， 50， 25 mg/kg， by raw material） after being stratified by body weight， with 10 rats in each group. Except for the normal group， the other groups were induced to construct GN model by giving 1 500 mg/kg potassium oxazinate and 100 mg/kg adenine intragastrically for 14 days. After modeling， the rats in each group were given relevant medicine/water intragastrically， once a day， for consecutive 28 days. After the last medication， the levels of biochemical parameters related to renal function [uric acid， creatinine （Cr）， blood urea nitrogen （BUN）， xanthine oxidase （XOD）] were detected in rats， and the histopathological changes in the rat kidney were observed. The protein expressions of monocyte chemoattractant protein-1（MCP-1），TNF-α and interleukin-6（IL-6） as well as the phosphorylation levels of p38 MAPK and NF-κB p65 protein were determined in renal tissue of rats. RESULTS Compared with the normal group， the model group suffered from the dilatation of renal tubules， structural damage to glomeruli， accompanied by inflammatory infiltration and fibrosis； the contents of uric acid， Cr， BUN and XOD， the protein expressions of MCP-1，TNF-α and IL-6 and the phosphorylation levels of p38 MAPK and NF-κB p65 protein were all increased significantly （P＜0.05 or P＜0.01）. Compared with the model group， the pathological symptoms of renal tissue in rats had been improved to varying degrees in different dose groups of PAP； the contents of uric acid， Cr， BUN and XOD， protein expressions of MCP-1， TNF-α and IL-6， the phosphorylation levels of p38 MAPK and NF-κB p65 protein in PAP high-dose and PAP medium-dose groups were all decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS PAP exhibits an anti-GN effect， the mechanism of which may be associated with inhibiting the p38 MAPK/NF-κB/TNF-α signaling pathway.

Latest research and prospect of CD47 in kidney transplantation / 器官移植

CD47 is a transmembrane protein widely expressed on cell surface, which is considered as a key molecule for immune escape. With an increasing number of related studies, the role of CD47 and its ligands in immunomodulatory effects has been gradually understood. Recent studies have investigated the role of CD47 in ischemia-reperfusion injury of allogenetic kidney transplantation, rejection and xenotransplantation. Nevertheless, the specific role and the key mechanism remain elusive. In this article, the structure and function of CD47, common CD47 ligands, the relationship between CD47 and kidney transplantation, and the application of CD47 in kidney transplantation were reviewed, the latest research progress of CD47 in kidney transplantation was summarized, and the limitations of current research and subsequent research direction were analyzed, aiming to provide reference for subsequent application of CD47 in allogeneic and kidney xenotransplantation.

Liver transplantation for end-stage hepatic alveolar echinococcosis / 器官移植

Hepatic alveolar echinococcosis (HAE) is a common zoonotic endemic parasitic disease in western China. It lacks of typical clinical manifestations in the early stage, and symptoms become prominent during the end stage, with an alarmingly high mortality rate. Among the treatment of end-stage HAE (es-HAE), orthotopic liver transplantation is almost the only radical treatment due to insufficient remnant liver volume, uncontrollable bleeding and difficulty in vascular reconstruction in vivo. However, the shortage of donor liver and long-term postoperative use of immunosuppressants limit its application. The introduction of ex vivo liver resection and autotransplantation (ELRA) resolves this dilemma and significantly broadens the indications of es-HAE. In addition, multiple centers in China have optimized and modified ELRA to further improve the treatment system of es-HAE. At present, liver transplantation (including ELRA) of es-HAE remains a hot topic for clinicians. In this article, orthotopic liver transplantation, ELRA, auxiliary ELRA and other surgical treatment of es-HAE were reviewed, aiming to further enhance the diagnosis and treatment of es-HAE and improve clinical prognosis of the patients.

Combination of AAV-delivered tumor suppressor PTEN with anti-PD-1 loaded depot gel for enhanced antitumor immunity

Recent clinical studies have shown that mutation of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene in cancer cells may be associated with immunosuppressive tumor microenvironment (TME) and poor response to immune checkpoint blockade (ICB) therapy. Therefore, efficiently restoring PTEN gene expression in cancer cells is critical to improving the responding rate to ICB therapy. Here, we screened an adeno-associated virus (AAV) capsid for efficient PTEN gene delivery into B16F10 tumor cells. We demonstrated that intratumorally injected AAV6-PTEN successfully restored the tumor cell PTEN gene expression and effectively inhibited tumor progression by inducing tumor cell immunogenic cell death (ICD) and increasing immune cell infiltration. Moreover, we developed an anti-PD-1 loaded phospholipid-based phase separation gel (PPSG), which formed an in situ depot and sustainably release anti-PD-1 drugs within 42 days in vivo. In order to effectively inhibit the recurrence of melanoma, we further applied a triple therapy based on AAV6-PTEN, PPSG@anti-PD-1 and CpG, and showed that this triple therapy strategy enhanced the synergistic antitumor immune effect and also induced robust immune memory, which completely rejected tumor recurrence. We anticipate that this triple therapy could be used as a new tumor combination therapy with stronger immune activation capacity and tumor inhibition efficacy.

Spatial transcriptomics reveals that metabolic characteristics define the tumor immunosuppression microenvironment via iCAF transformation in oral squamous cell carcinoma / 国际口腔科学杂志·英文版

Tumor progression is closely related to tumor tissue metabolism and reshaping of the microenvironment. Oral squamous cell carcinoma (OSCC), a representative hypoxic tumor, has a heterogeneous internal metabolic environment. To clarify the relationship between different metabolic regions and the tumor immune microenvironment (TME) in OSCC, Single cell (SC) and spatial transcriptomics (ST) sequencing of OSCC tissues were performed. The proportion of TME in the ST data was obtained through SPOTlight deconvolution using SC and GSE103322 data. The metabolic activity of each spot was calculated using scMetabolism, and k-means clustering was used to classify all spots into hyper-, normal-, or hypometabolic regions. CD4T cell infiltration and TGF-β expression is higher in the hypermetabolic regions than in the others. Through CellPhoneDB and NicheNet cell-cell communication analysis, it was found that in the hypermetabolic region, fibroblasts can utilize the lactate produced by glycolysis of epithelial cells to transform into inflammatory cancer-associated fibroblasts (iCAFs), and the increased expression of HIF1A in iCAFs promotes the transcriptional expression of CXCL12. The secretion of CXCL12 recruits regulatory T cells (Tregs), leading to Treg infiltration and increased TGF-β secretion in the microenvironment and promotes the formation of a tumor immunosuppressive microenvironment. This study delineates the coordinate work axis of epithelial cells-iCAFs-Tregs in OSCC using SC, ST and TCGA bulk data, and highlights potential targets for therapy.

Clinical characteristics and treatment analysis of three cases of congenital ulnar collateral flexor contracture of the forearm / 中国修复重建外科杂志

OBJECTIVE@#To report the clinical characteristics and treatment analysis of 3 cases of congenital ulnar collateral flexor contracture of the forearm and take a reference for clinic.@*METHODS@#A total of 3 patients with congenital ulnar collateral flexor contracture of the forearm were admitted between February 2019 and August 2021. Two patients were male and 1 was female, and their ages were 16, 20, and 16 years, respectively. The disease durations were 8, 20, and 15 years, respectively. They all presented with flexion deformity of the proximal and distal interphalangeal joints of the middle, ring, and little fingers in the neutral or extended wrist position, and the deformity worsened in the extended wrist position. The total action motion (TAM) scores of 3 patients were 1 and the gradings were poor. The Carroll's hand function evaluation scores were 48, 55, and 57, and the grip strength indexes were 72.8, 78.4, and 30.5. Preoperative CT of case 2 showed a bony protrusion of the flexor digitorum profundus tendon at the proximal end of the ulna; and MRI of case 3 showed that the ulnar flexor digitorum profundus presented as a uniform cord. After diagnosis, all patients were treated with operation to release the denatured tendon, and functional exercise was started early after operation.@*RESULTS@#The incisions of 3 patients healed by first intention. Three patients were followed up for 12, 35, and 12 months, respectively. The hand function and the movement range of the joints significantly improved, but the grip strength did not significantly improve. At last follow-up, TAM scores were 3, 4, and 4, respectively, among which 2 cases were excellent and 1 case was good. Carroll's hand function evaluation scores were 95, 90, and 94, and the grip strength indexes were 73.5, 81.3, and 34.2, respectively.@*CONCLUSION@#Congenital ulnar collateral flexor contracture is a rare clinical disease that should be distinguished from ischemic muscle contracture. The location of the contracture should be identified and appropriate surgical timing should be selected for surgical release. Active postoperative rehabilitation and functional exercise can achieve good hand function.

Prognostic value of CC motif chemokine receptor 2 in the microenvironment of glioblastoma and its relationship with the immune cell infiltration / 肿瘤研究与临床

Objective:To explore the prognostic biomarkers of glioblastoma (GBM) in the tumor microenvironment (TME) and its function.Methods:A total of 169 GBM samples of 161 GBM patients were collected from the Cancer Genome Atlas (TCGA) database. ESTIMATE algorithm in R4.1.0 software was used to calculate the proportion of immune components and stromal components in TME, which were expressed as immune score and stromal score, respectively. According to the median value of the two scores, 169 GBM samples were divided into the high score group and the low score group, respectively, 84 each in each group (those whose scores were equal to the median were not involved in the grouping). The differentially expressed genes (DEG) [false discovery rate (FDR) < 0.05] between the high score group and the low score group of the two scores were obtained by using limma package, and the co-up-regulated and co-down-regulated DEG of the two scores were obtained by using Venn program. Based on the STRING database, the protein interaction (PPI) network of co-up-regulated and down-regulated DEG of immune score and stromal score was constructed, and the top 30 genes with connectivity were selected. Univariate Cox proportional hazard model analysis of overall survival (OS) of 161 GBM patients in the TCGA database was performed on co-up-regulated and down-regulated DEG between immune score and stromal score by using R4.1.0 software to obtain the DEG affecting OS. The intersection of the DEG obtained from PPI analysis and Cox analysis was taken as the prognostic core genes. According to the median expression value of prognostic core genes in GBM samples from the TCGA database, 161 patients were divided into prognostic core genes high expression group and low expression group (patients whose scores were equal to the median were not involved in the grouping), with 80 cases in each group. Kaplan-Meier survival analysis of OS was performed by using R4.1.0 software. GSEA 4.2.1 software was used to perform gene set enrichment analysis (GSEA) on all genes with transcriptome data of GBM patients in the two groups of the TCGA databases, and the main enriched functions of the two groups of genes were obtained. The CIBERSORT algorithm was used to test the accuracy of the proportion of tumor infiltrating immune cell (TIC) subsets in 169 GBM samples from the TCGA database, and 57 GBM samples were finally obtained. Immune cells with differential expression levels and immune cells related to the expression of prognostic core genes among the samples with different expression levels of prognostic core genes were analyzed; Venn program was used to obtain the intersection of immune cells with differential levels and related immune cells, and differentially expressed TIC related to expressions of prognostic core genes in GBM were obtained.Results:Based on the immune score and stromal score of GBM samples in the TCGA database, a total of 693 co-up-regulated and co-down-regulated DEG of both scores were screened out. After the intersection of 78 DEG related to OS obtained by univariate Cox regression analysis and 30 DEG obtained by PPI network results, CC motif chemokine receptor 2 (CCR2) was identified as the prognostic core gene ( HR = 1.294, 95% CI 1.060-1.579, P = 0.011). GBM patients with CCR2 high expression had worse OS compared with those with CCR2 low expression ( P = 0.009). GSEA analysis showed that genes in the CCR2 high expression group were mainly enriched in immune-related pathways, while genes in the CCR2 low expression group were mainly enriched in metabolism-related pathways. Among 57 screened GBM samples, there were differences in the levels of 3 immune cells between the CCR2 high expression group and the CCR2 low expression group ( P < 0.05). CCR2 expression was correlated with the levels of 9 immune cells (all P < 0.05). Venn program analysis showed that differentially expressed 3 TIC in GBM related to CCR2 gene expression were obtained; among them, M2 macrophages were positively correlated with CCR2 expression, while T follicular helper cell and activated NK cells were negatively correlated with CCR2 expression. Conclusions:CCR2 may be the core gene related to the prognosis in the TME of GBM. As reference, the level of CCR2 can help to predict the status of TME and prognosis in GBM patients, which is expected to provide a new direction for the treatment of GBM.

Analysis and thoughts on the settings of sub-campus of public hospitals directly under the National Health Commission of China / 中华医院管理杂志

Objective:To analyse the settings of sub-campus of public hospitals directly under the National Health Commission, so as to provide research reference and suggestions for further promoting the scientific and standardized construction of sub-campuses of public hospitals.Methods:Data before August 30, 2022 were collected from the official websites of the hospitals in question including their introduction, overview, navigation, news and other categories of information.Relevant literatures on multi-campus published by hospitals under the direct management of the National Health Commission were obtained from the China National Knowledge Network, which was identified by searching the keywords " sub or multi-campus hospital" with " author affiliation", using such search conditions as " MeSH major topic" or " title/keywords/abstract" of the studied hospitals in advanced search builder. Descriptive analysis and frequency analysis were conducted on the numbers, the construction reasons, geographical location, operation scale, characteristics and positioning of the sub-campuses in the studied hospitals.Results:Of the 44 such hospitals, 30 had a total of 50 sub-campuses in operation. Generally there were two modes of construction. 35 were newly constructed and 15 formed by mergers; 37 sub-campuses were built and put into operation since 2010. Sub-campuses were usually located in the same area of the main campus, with 11 in the same area of the same city and areas, 35 in different areas of the same city, and 4 in different cities; the average geographical distance between the sub-campus and the main hospital campus was 31.18 kilometers; 27 sub-campuses had beds ≤ 500, 12 sub-campuses had beds between 501-1 000, 11 had beds>1 000; 14 specialized sub-campuses and 31 out of the 35 comprehensive sub-campuses clearly indicating their characteristics of specialized departments, and 1 other (research institute areas with research beds).Conclusions:Future proofing is characteristic of these public hospitals in terms of their sub-campus construction, regional setting, planning and positioning, and functional layout. However, both government regulatory departments and public hospitals should comprehensively consider and make prudent decisions on the planning, investment, demonstration, layout, and other aspects of the construction of sub-campuses of hospitals.

Application of MassARRAY genotyping assay in the diagnosis of neonatal genetic metabolic diseases / 中华检验医学杂志

Objective:To investigate the accuracy, effectiveness and feasibility of MassARRAY genotyping assay in the diagnoses of neonatal genetic metabolic diseases.Methods:This is a retrospective study. From December 2016 to January 2020, newborns were screened by tandem mass spectrometry at the Zhejiang Newborn Screening Center, among which the data of 7 922 suspected positive cases of genetic metabolic diseases were collected. These patients were then tested for the common variants of 27 genetic metabolic diseases by MassARRAY genotyping assay, along with further testing using Sanger or next-generation sequencing used to verify and/or further search for potential variants.Results:A total of 1 408 cases were tested with MassARRAY. Among these, 307 cases were confirmed with certain genetic metabolic diseases. The detection rate of hyperphenylalaninemia was the highest, followed by primary carnitine deficiency, short acyl-coA dehydrogenase deficiency and methylmalonic acidemia. With these cases, the consistency of Sanger sequencing and MassARRAY was 100% (307/307). Another 287 cases were identified as carriers by MassARRAY with a 49.1% (141/287) consistency in reference to Sanger sequencing, mainly involving SLC22A5 and MCCC1 genes. Meanwhile, 50.8% (146/287) of these cases were found to have another variant mainly involving PAH, PTS and ACADS genes. The remaining 814 cases have no variants; 158 cases out of these patients have continuously abnormal amino acids, acyl carnitines, urine organic acid and/or other biochemical indices, and were tested by next-generation sequencing, among which 38% (60/158) were detected with two variants. In this study, a total of 513 patients with genetic metabolic disease were diagnosed, and the detection rate of MassARRAY was 59.8% (307/513). Conclusions:MassARRAY genotyping assay can be used as an early molecular screening method for neonatal genetic metabolic diseases. The detection rate is particularly high in diseases with a high concentration of hotspot variants, such as hyperphenylalaninemia and primary carnitine deficiency. The future application value of MassARRAY should be further improved by continuously optimizing its ability to identify new disease genes and potential variable sites.

A survey on clinical practice of digestive endoscopy in county-level hospitals of Yunnan Province / 中华消化内镜杂志

Objective:To analyze general information, diagnosis and treatment of digestive endoscopy departments in county-level hospitals of Yunnan Province, thus to provide evidence for improving the endoscopic diagnosis and therapy in these hospitals.Methods:An online survey was performed to collect quality-related information of digestive endoscopy centers (departments) at county-level hospitals of Yunnan from January 2019 to January 2020, including endoscopic equipment, endoscopic techniques, staffing, number of operations, and quality control.Results:A total of 143 county-level hospitals were involved in this study. Each hospital owned only 1.74 endoscopy operation rooms on average, 1.42 regular endoscopy workstation, 4 endoscopes. There were only 10 endoscopic ultrasonography workstations and 2 enteroscopy workstations respectively installed in these hospitals. Endoscopic retrograde cholangiopancreatography was independently performed in only 4 hospitals. There were 392 professional endoscopists in all 143 hospitals, 18.62% (73/392) of whom were able to perform endoscopic therapy of grade 4, while only 6.12% (24/392) of whom could perform endoscopic submucosal dissection (ESD) independently. In 2019, the early diagnostic rate of gastrointestinal tract cancer in these hospitals was 19.48% (1 133/5 817). The early diagnostic rate was 21.04% (276/1 312) for esophageal cancer, 19.53% (397/2 033) for gastric cancer, and 18.61% (460/2 472) for colorectal cancer. The colorectal adenoma detection rate was 12.83% (12 207/95 148). The rate of reaching cecum during colonoscopy was 70.49% (67 067/95 148). The complete resection rate of ESD was 12.54% (221/1 763).Conclusion:The general situation of digestive endoscopy in county-level hospitals of Yunnan is far from being well-developed, which may be attributed to slow upgrading of equipments, inadequate training of endoscopic professionals and undemanding quality control. It is highly significant to improve medical service and quality of diagnosis and treatment of digestive endoscopy in these hospitals, on the basis of a complete endoscopy quality control system with a high standard.

Association between pregnancy-related factors and stillbirth: a retrospective cohort study based on 500 000 pregnant residents in Wuhan / 中华围产医学杂志

Objective:To analyze the incidence of stillbirth and the associated factors in pregnancy among pregnant residents in Wuhan.Methods:A previous birth cohort was retrospectively reviewed. The cohort was based on Wuhan Maternal and Child Information System, and the perinatal information of pregnant residents in Wuhan from January 1, 2011, to September 30, 2017 and information of selected cases was collected, including socio-demographic characteristics, pregnant history, and healthcare information during pregnancy and labor. Data on stillbirth, including fetal death in uterus and in labor, were selected for this study. Chi-square test was adopted for comparing the differences in pregnancy-related factors between live birth and stillbirth, and binary logistic regressions for exploring the influencing factors associated with the occurrence of stillbirth. Results:A total of 509 057 deliveries in Wuhan were included in this study, including 505 839 live births and 3 218 stillbirths (3 155 after exclusion of fetal death in labor), with an overall incidence of stillbirth of 6.32‰(3 218/509 057), and an annual incidence between 4.90‰ to 8.11‰. Statistically significant differences were found between the live birth and stillbirth group in the following items: maternal age [<25 years old: 19.28% (97 544/505 839) vs 19.36% (623/3 218); 25-30 years old: 48.45% (245 077/505 839) vs 45.15% (1 453/3 218); 30-35 years old: 26.09% (131 952/505 839) vs 26.29% (846/3 218); >35 years old: 6.18% (31 266/505 839) vs 9.20% (296/3 218)], educational background [middle school or below: 22.90% (115 833/505 839) vs 22.03% (709/3 218); high school: 36.37% (183 978/505 839) vs 38.72% (1 246/3 218); college or above: 40.73% (206 028/505 839) vs 39.25% (1 263/3 218)], occupation [brainworker or professionals: 33.51% (169 514/505 839) vs 31.54% (1 015/3 218); manual or freelance worker: 66.38% (335 763/505 839) vs 68.34% (2 199/3 218)], residential area [urban area: 70.00% (354 365/505 839) vs 76.32% (2 456/3 218); rural area: 30.00% (151 474/505 839) vs 23.68%(762/3 218)], and time of conception [spring (March to May): 24.27% (122 746/505 839) vs 24.08% (775/3 218); summer (June to August): 24.09% (121 867/505 839) vs 23.87% (768/3 218); fall (September to November): 26.69% (135 012/505 839) vs 25.08% (807/3 218); winter (December to next February): 24.95% (126 214/505 839) vs 26.97% (868/3 218)] (all P<0.05), but no significant difference was found in fetal gender ( P>0.05). Besides, gravidity [once: 49.32% (249 484/505 839) vs 47.02% (1 513/3 218); over twice: 50.68% (256 355/505 839) vs 52.98% (1 705/3 218)], parity [once: 73.60% (372 316/505 839) vs 77.07% (2 480/3 218); over twice: 26.40% (133 523/505 839) vs 22.93% (738/3 218)], history of stillbirth [0.33% (842/256 355) vs 0.65% (11/1 705)], hypertensive disorders in pregnancy [3.25% (16 464/505 839) vs 5.59% (180/3 218)], first trimester vaginal bleeding [2.02% (10 251/505 839) vs 2.61% (84/3 218)], placenta previa [0.98% (4 963/505 491) vs 2.64% (53/2 009)], and oligohydramnios [2.52% (12 764/505 839) vs 1.90% (61/3 218)] differed significantly between the two groups (all P<0.05). However, no significant difference was found between the two groups in terms of the proportion of women with gestational diabetes mellitus and previous spontaneous abortion (both P>0.05). After exclusion of fetal death in labor from the 3 218 stillbirths, the same results were achieved. Binary logistic regression analysis showed that women who were over 30 years old (30-35 years old: OR=1.42, 95% CI: 1.30-1.56; >35 years old: OR=2.59, 95% CI: 2.25-2.98), with a high school degree or below (middle school or below: OR=1.37, 95% CI: 1.21-1.55; high school: OR=1.28, 95% CI: 1.16-1.42), manual or freelance worker ( OR=1.18, 95% CI: 1.07-1.31), in the urban area ( OR=1.43, 95% CI:1.31-1.57), and gravidity ≥ 2 times ( OR=1.32, 95% CI: 1.21-1.43), primiparity ( OR=1.76, 95% CI: 1.58-1.96), gestational hypertension ( OR=2.80, 95% CI: 2.40-3.27), vaginal bleeding in the first trimester ( OR=1.35, 95% CI: 1.08-1.68), placenta previa ( OR=10.86, 95% CI: 8.84-13.35) and history of stillbirth ( OR=2.27, 95% CI: 1.30-3.98) were all risk factors of stillbirth. Conclusion:Pregnant women who were over 30 years old, less educated, manual worker or freelance or with a history of adverse pregnancy, vaginal bleeding in the first trimester, hypertension in pregnancy, and placenta previa are at higher risk of stillbirth

Study on safety performance and condition-suggestion accuracy of the symptom assessment mobile applications / 中华全科医师杂志

Objective:To compare the breadth of condition coverage, accuracy of suggested conditions and appropriateness of urgency advice of the 8 symptom assessment mobile applications (APPs) available on the Chinese market.Methods:The APPs were assessed using 200 primary care vignettes and were measured against the vignettes′ standard. The primary outcome measures were proportion of conditions covered by an APP, proportion of vignettes with the correct primary diagnosis,and proportion of safe urgency advice.Results:For APPs assessed,condition-coverage was from 29.0%(58/200)to 99.5%(199/200), top-3 suggestion accuracy was from 8.5%(17/200) to 61.5%(123/200), the proportion of safe urgency advice was from 84.8%(167/197) to 99.5% (198/199).Conclusions:The APPs showed a wide range of coverage, safety performance and condition-suggestion accuracy. Symptom assessment APPs with good performance could be used by general practitioners as supporting tools. However, even symptom assessment APPs with excellent performance need to be further assessed in a real clinical environment.

Role of NALCN in hippocampal dentate gyrus in social behavior of mice / 中华麻醉学杂志

Objective:To evaluate the role of the sodium leak channel (NALCN) in the hippocampal dentate gyrus (DG) in the social behavior of mice.Methods:Thirty-nine male wild-type C57BL/6 mice, aged 6-8 weeks, weighing 18-22 g, were used in this study. Three mice were sacrificed to verify the expression and co-expression of NALCN with neuronal nuclear antigen (NeuN) in the hippocampal DG using the immunofluorescent staining. The remaining 36 mice were divided into 2 groups ( n=18 each) by the random number table method: control group (group C) and NALCN gene knockdown group (group KO). NALCN-shRNA virus was injected in group KO, and scrambled-shRNA virus was injected in group C. The three box social test and open field test were performed at 3 weeks after the virus injection. Mice were sacrificed under anesthesia after the behavioral test, hippocampal tissues were collected, and the injection location of the virus was verified with a fluorescence microscope, and the NALCN protein and mRNA expression in the hippocampal DG was detected by Western blot and real-time polymerase chain reaction, respectively. Results:NALCN and NeuN co-expressed a lot on the same neuron in the hippocampal DG of mice, indicating that NALCN was widely expressed on the neurons in the hippocampal DG. Compared with group C, the expression of NALCN and mRNA in the hippocampal DG was significantly down-regulated, and the social novelty preference disappeared ( P<0.05), and no significant change was found in the social ability and each parameter in the open field test in group KO ( P>0.05). Conclusions:NALCN in the hippocampal DG is involved in the regulation of social memory in mice, and the down-regulated expression of NALCN can lead to the loss of social novelty preference in mice.