Objective To analyze the clinical efficacy of total laparoscopic proximal gastrectomy and esophagogastric anastomosis to provide a new surgical method for the treatment of early proximal gastric cancer. Methods The clinical data of 80 patients with early gastric cancer who underwent proximal gastrectomy in the Department of General Surgery, the First Medical Center of Chinese PLA General Hospital from January 2019 to June 2021 were retrospectively analyzed. According to different surgical methods, the patients were divided into two groups: a total laparoscopic proximal gastrectomy group (n=36) and a laparoscopic-assisted proximal gastrectomy group (n=44). The perioperative conditions, long-term complications, and survival status were compared between the two groups. Results The length of surgical incision [(59.9±4.7) mm vs. (119.7±8.3) mm, P<0.001], first exhaust time [(58.2±15.3) h vs. (66.8±16.4) h, P=0.019] and postoperative hospital stay [(7.6±1.1) d vs. (9.2±1.3) d, P<0.001] of total laparoscopic proximal gastrectomy group were significantly shorter than those of laparoscope-assisted proximal gastrectomy group. The duration of operation [(186.9±16.4) min vs. (154.0±17.2) min, P<0.001] of total laparoscopic proximal gastrectomy group was significantly longer than that of laparoscopic-assisted proximal gastrectomy group. There were no significant differences in intra-operative hemorrhage, number of lymph nodedissection and first liquid diet feeding time between the two groups (P>0.05), and no early complications requiring surgical intervention occurred. The incidence of reflux esophagitis in the total laparoscopic proximal gastrectomy group was less than that in laparoscope-assisted proximal gastrectomy group [16.7%(6/36) vs. 38.6%(17/44), P=0.031]. There was no significant difference in gastric motility, residual gastritis, anastomosis stenosis and anastomotic ulcer between the two groups (P>0.05). Conclusion Total laparoscopic proximal gastrectomy and esophagogastric anastomosis have the characteristics of good safety, rapid postoperative recovery and anti-reflux, being worthy of clinical popularization.

Objective To explore the application value of pathogen-targeted next-generation sequencing (tNGS) technology in patients with suspected pulmonary infections. Methods A retrospective analysis was conducted on the clinical data of 80 patients with suspected pulmonary infections admitted to the Department of Respiratory and Critical Care Medicine at the Third People's Hospital of Hubei Province from January 2021 to July 2023. All patients underwent bronchoalveolar lavage fluid (BALF) tNGS and conventional pathogen detection. Demographic characteristics of the patients were analyzed, and the distribution of pathogens detected by tNGS and conventional methods were compared. The clinical data of patients diagnosed with single pulmonary infections and those with mixed infections were also compared. Results Among the 80 patients, 74 were diagnosed with infections. Most of the infected patients had underlying conditions, mainly chronic heart disease (42.5%), chronic respiratory disease (35%), and diabetes (20%). The tNGS test results led to changes in treatment strategy for 35 patients (43.8%). A total of 45 types of pathogens were detected, with 169 strains identified by tNGS and 63 strains by conventional methods. Within pathogens detected by both methods, bacteria were detected the most. The order of pathogen types detected by tNGS was bacteria > viruses > fungi > atypical pathogens > Mycobacterium tuberculosis. The order of pathogen types detected by conventional methods was fungi > viruses > bacteria > atypical pathogens > Mycobacterium tuberculosis. The consistency between the two pathogen detection methods was poor (kappa value 0.172, P=0.020). The number of positive cases and the positive detection rates for bacteria, viruses, and atypical pathogens detected by tNGS were significantly higher than those of conventional methods (P<0.05), but there was no statistically significant difference in the positive detection rates for fungi and Mycobacterium tuberculosis between the two methods (P>0.05). Using clinical diagnosis as the gold standard, the sensitivity of tNGS detection was significantly higher than that of conventional methods (P=0.026), while there was no statistically significant difference in specificity between the two methods (P>0.05). Among the 74 confirmed pulmonary infection cases, 6 had no clear pathogen, 23 had single infections, and 45 had mixed infections. Among the mixed infections, the most common combination was bacterial-viral mixed infections (12/45, 26.7%). The mortality rate and hospitalization duration of patients with mixed infections were significantly higher than those with single infections (P<0.05); there were no statistically significant differences in gender, age, underlying conditions, white blood cell count, and neutrophil percentage between the two groups (P>0.05). Conclusions tNGS technology has higher pathogen detection sensitivity compared to conventional methods, especially for bacteria, viruses, atypical pathogens, and rare pathogens. This technology is beneficial for identifying mixed infections and can serve as a supplement to conventional pathogen detection methods in clinical practice.

Objective To investigate the effect of Mex3c gene knockout on embryonic neural tube development and its possible mechanisms. Methods The NCBI database was used to analyze the expression of Mex3c gene in various tissues of mice. Fluorescence in situ hybridization (FISH) was employed to detect the expression of Mex3c in neural tubes of Mex3c^+/+ mice at different developmental stages (E12.5 d, E14.5 d). Sexual mature mice were mated at a ratio of Mex3c^+/- male to female (1:1) in the same cage. Embryos were collected and genotyped using PCR. They were divided into 3 groups based on their genotype: wild-type group (Mex3c^+/+, WT group), homozygous knockout group (Mex3c^-/-, KO group), and heterozygous knockout group (Mex3c^+/-). HE staining was employed to observe the development of neural tubes in the 3 groups of embryos. Immunofluorescence staining and Western blotting were performed to detect the proliferation and apoptosis of embryonic neural stem cells in the WT and KO groups. Transmission electron microscopy was used to observe the ultrastructure of the neural tubes and mitochondria in the WT and KO groups. RNA was extracted from the neural tubes of WT and KO groups for RNA-seq sequencing. The R.3.6.3 software was used to perform KEGG signaling pathway enrichment analysis on differentially expressed genes. RT-qPCR was used to validate the sequencing results. Results The NCBI database analysis and FISH detection results showed that the Mex3c gene was mainly expressed in the central nervous system of embryos. HE staining results showed that there was no significant difference in the development of embryonic neural tubes between KO group, WT group, and heterozygous knockout group at E12.5 d and E13.5 d. However, at E14.5 d, the embryonic neural tube development in KO group was delayed and the phenotype was significantly abnormal compared with those in WT group. Therefore, the embryonic neural tube tissues of KO group and WT group at E14.5 d were selected for subsequent experiments. The immunofluorescence staining results showed that the PCNA positive cell rate in KO group was significantly lower than that in WT group (P<0.001). The Western blotting results showed that the Bax/Bcl-2 ratio in KO group was higher than that in WT group (P<0.01). Transmission electron microscopy observation showed that compared with WT group, the synaptic gap in KO group disappeared, the mitochondrial of the embryonic neural tube in KO group were swollen, the mitochondrial cristae were disrupted, and the structure was significantly abnormal. The results of RNA-seq analysis showed that a total of 377 differentially expressed genes were obtained, including 101 up-regulated genes and 276 down-regulated genes. KEGG signaling pathway enrichment analysis revealed that the main signaling pathways of differentially expressed genes were enriched in the neuroactive ligand receptor interaction signaling pathways. The RT-qPCR validation results showed that the mRNA expression levels of Avpr1a, Drd1, Htr7, Sstr1, Oxtr and Gabra5 in this signaling pathway were down-regulated (P<0.05 or P<0.01), which was consistent with the RNA-seq results. Conclusion Mex3c plays an important role in the development of neural tubes in mouse embryos, which may participate in regulating the proliferation and apoptosis of neural stem cells through neural active ligand receptor interaction signaling pathways, thereby affecting the development of neural tubes.

Objective To compare the fidelity of chronic obstructive pulmonary disease (COPD) models established using two methods: exposure to cigarette smoke (CS) and exposure to motor vehicle exhaust (MVE) in rats. Methods Twenty-four male SD rats were randomly divided into control, CS-exposed (CS), and MVE-exposed (MVE) groups, with 8 rats per group. Rats in CS and MVE groups were exposed to CS or MVE, respectively, to induce COPD models. After COPD model established, lung function of each group was assessed. Bronchoalveolar lavage fluid (BALF) was collected to measure inflammatory cell counts, levels of inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor (TNF)‑α, and expression levels of mucin 5AC (MUC5AC). Lung tissue sections were stained with hematoxylin and eosin (HE) to observe pulmonary tissue and airway pathological changes. Periodic acid-Schiff (PAS) staining was used to detect goblet cell hyperplasia in airways. Results Compared with control group, rats in CS and MVE groups showed significantly increased inspiratory resistance (RI), total lung capacity (TLC), and lung static compliance (Cchord) (P<0.05), while expiratory flow parameters FEV₅₀/FVC were significantly decreased (P<0.05). Compared with MVE group, rats in CS group had significantly higher R_I, TLC, and Cchord (P<0.05), and lower FEV₅₀/FVC (P<0.05). HE staining of lung tissues showed that mean linear intercept (MLI) was significantly higher in both CS and MVE groups compared with control group (P<0.05), with CS group having higher MLI than MVE group (P<0.05). BALF analysis revealed that white blood cells, neutrophils, macrophages, lymphocytes, IL-6, and TNF-α levels were significantly higher in both CS and MVE groups compared with control group (P<0.05), and inflammatory cell counts, IL-6, and TNF-α levels were higher in CS group compared with MVE group(P<0.05). PAS staining of lung tissues indicated that goblet cells in large airways were significantly increased in both CS and MVE groups compared with control group (P<0.05), with CS group showing higher goblet cell counts than MVE group (P<0.05). Expression levels of MUC5AC in BALF were significantly higher in both CS and MVE groups compared with control group (P<0.05), with CS group having significantly higher MUC5AC levels than MVE group (P<0.05). Conclusions Exposure to CS or MVE can establish a rat model of COPD, with CS exposure better mimicking characteristics of acute exacerbation of COPD compared to MVE exposure.

Objective To analyze the effects and mechanisms of a disintegrin and metalloproteinase with thrombospondin motifs 14 (ADAMTS14) gene in gastric cancer associated with Helicobacter pylori (Hp) infection and explore the potential of ADAMTS14 as a novel target for biological therapy of gastric cancer. Methods The Caner Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases were utilized to analyze differentially expressed genes in gastric cancer tissues and Hp-positive gastric mucosa, and to screen potential targets. For the selected target ADAMTS14, its expression pattern in gastric cancer and Hp-positive gastric mucosa was analyzed. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis was performed on the genes co-expressed with ADAMTS14 in gastric cancer. Kaplan-Meier Plotter database was used to analyze the correlation between high- and low-ADAMTS14 expression and prognosis of gastric cancer patients. The expression of ADAMTS14 and its relationship with clinicopathological features in 30 patients with gastric cancer were analyzed using real-time fluorescent quantitative PCR (RT-qPCR) and immunohistochemistry. The relationship between Hp infection and ADAMTS14 expression was analyzed using GEO database, and confirmed by cell line infection experiment. ADAMTS14 expression in gastric cancer cell line NCI-N87 was silenced by small interfering RNA to analyze its effect on the function of gastric cancer cells; The effect of ADAMTS14 knockdown on downstream target genes of Hippo signaling pathway was analyzed by RT-qPCR and Western blotting. Results A total of 16 genes closely related to Hp infection and gastric cancer were identified by analyzing the TCGA and GEO databases. According to TCGA database, the ADAMTS14 expression in gastric cancer tissues was higher than that in healthy gastric mucosa (P<0.0001). RT-qPCR and immunohistochemistry results also showed higher ADAMTS14 expression in gastric cancer tissues compared to adjacent tissues (P<0.01). Survival analysis demonstrated poor prognosis in patients with high ADAMTS14 expression (P<0.001). Data from GSE60427 showed that ADAMTS14 expression was upregulated in Hp-infected gastric mucosa and cell lines (P<0.001). After Hp P12 infection, the ADAMTS14 expression in NCI-N87 cells was higher than that in uninfected group. ADAMTS14 knockdown significantly inhibited the proliferation and migration of NCI-N87 cells (P<0.05), and it also significantly inhibited the expression of Hippo signaling pathway target genes, including BMP7, BMPR2, FZD4, PARD3 and SAV1 (P<0.05). Conclusion ADAMTS14 is highly expressed in Hp-positive gastric mucosa and gastric cancer tissues, which may accelerate gastric cancer progression by regulating Hippo signaling pathway, and holds potential as a new marker and therapeutic target for Hp infective gastric cancer.

Objective To investigate the effect of dexmedetomidine (Dex) on the rat model of perioperative stroke and its mechanism. Methods One hundred male rats were randomly divided into sham group , middle cerebral artery occlusion (MCAO) group, low dose Dex [Dex-L, 0.5 μg/(kg·h)] group, medium dose Dex [Dex-M, 2 μg/(kg·h)] group, high dose Dex [Dex-H, 10 μg/(kg·h)] group, 20 rats in each group. A rat model of perioperative stroke was established by middle cerebral artery occlusion. Dex was injected intravenously at different doses during ischemia. After 24 h, the neurological function of the rats was evaluated. Then, the rats were sacrificed and the peripheral blood and whole brain tissue were collected, ischemic core area tissue was separated from some brain tissues and the cerebral infarction area was observed by TTC staining. The inflammatory cytokine contents in serum and ischemic core area were measured by ELISA. In addition, the expressions of formyl peptide receptor 1 (FPR1), transmembrane protein 119 (TMEM119), CD31 and VE-cadherin proteins were assayed by immunofluorescence, and the expressions of FPR1, nuclear factor-κB (NF-κB) and NLRP3 proteins by Western blotting in the ischemic core area. Results Compared with sham group, the proportion of cerebral infarction area and neurological scores in the MCAO group were significantly increased, and the contents of interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α in the serum and the ischemic core area were significantly increased, the expressions of FPR1, TMEM119, p-NF-κB, NLRP3, CD31 and VE-cadherin in the ischemic core were significantly increased(P<0.001), and there was obvious co-expression of FPR1 and TMEM119. Compared with MCAO group, the proportion of cerebral infarction area and neurological scores in the Dex-M and Dex-H groups were significantly decreased, and the contents of IL-1β, IL-6 and TNF-α in serum and brain ischemic core area were significantly decreased, the expressions of FPR1, TMEM119, p-NF-κB, NLRP3, CD31 and VE-cadherin in the ischemic core were significantly decreased (P<0.001). Conclusions Dex can significantly alleviate perioperative stroke injury. The mechanism may be due to inhibiting the expression of FPR1 protein, activation of microglia cells and cerebral collateral circulation angiogenesis.

Endoplasmic reticulum is an important organelle in eukaryotic cells, which is responsible for the folding, processing and transportation of secretory proteins. A variety of stimuli inside and outside cells can lead to the accumulation of misfolded or unfolded proteins in the endoplasmic reticulum, resulting in abnormal structure and function of the endoplasmic reticulum, which is called endoplasmic reticulum stress (ERS). Endoplasmic reticulum autophagy is an important endogenous mechanism to alleviate ERS. It is often considered as a cell protective procedure, which participates in many important physiological processes, such as metabolism, immune response, inflammatory response and cell proliferation. Endoplasmic reticulum autophagy is an important endogenous protective mechanism to alleviate endoplasmic reticulum stress and restore the endoplasmic reticulum homeostasis, through eliminating redundant and disabled endoplasmic reticulum membrane and macromolecular protein complexes, which is critical to cell function and fate. This paper reviews the types of endoplasmic reticulum autophagy, related specific receptors, main regulatory mechanisms, and its role and significance in the related diseases.

Post-stroke cognitive impairment (PSCI) is a prevalent functional impairments following stroke that seriously affects patients' quality of life and daily activities. Studies indicate a close relationship between intestinal microflora dysbiosis and central nervous system diseases. Intestinal microflora profoundly impacts on human physiological health, contributing to the stability of nervous, metabolic and immune systems through regulation of the gut-brain axis. An increasing number of studies confirmed the important role of the gut microbiome-gut-brain axis in the occurrence and development of stroke and its associated PSCI, and regulation of microbiome-gut-brain could be potential target to treatment of PSCI. This review summarizes research progress on gut microbiome-gut-brain axis and PSCI to provide a reference for exploration of related mechanisms and clinical prevention and treatment strategies.

With the in-depth study of molecular biology, non-small cell lung cancer (NSCLC) has opened the era of precision medicine based on mutation-based molecular targeting therapy. Epidermal growth factor receptor (EGFR) driver mutations are closely related to the progression of NSCLC, and EGFR-tyrosine kinase inhibitors (TKIs) developed based on this have achieved significant therapeutic effects, but acquired drug resistance is still one of the major factors limiting their long-term use. As resistance mechanisms are further investigated, in addition to secondary EGFR mutation, MET amplification, HER2 amplification, histologic transformation, etc., receptor tyrosine kinase (RTK) fusion mutation have been shown to be a targetable mechanism of acquired resistance. Among the acquired RTK fusion mutations, rearranged during transfection (RET) fusion mutations are the accessible targets of our concern. As the RET molecule continues to be explored, drugs targeting RET fusions have been approved and marketed. There are different clinical strategies to deal with acquired RET fusion mutation mediating resistance to EGFR-TKIs treatment. In this review, the structure and function of RET, its relationship with EGFR-TKIs resistance, and treatment strategies are reviewed to further improve patient survival outcomes.

Lipids, including fats (triglycerides) and lipoids (phospholipids and sterols), not only serve as an energy source for the body but also play a pivotal role throughout the reproductive process, particularly in the establishment and maintenance of early pregnancy. This encompasses the regulate of early embryonic development and uterine tolerance, and the facilitation of embryo implantation. Given the diversity of lipids, this review focuses on extensively studied lipid mediators such as polyunsaturated fatty acids, endocannabinoids, prostaglandins, lysophosphatidic acid, sphingolipids and steroid hormones. It systematically elaborates on the regulatory effects of fatty acid, phospholipid, and cholesterol metabolism on the formation of endometrial receptivity and embryo implantation, as well as the potential underlying mechanisms. The review aims to provide new insights and feasible intervention approaches for predicting and improving the outcomes of natural pregnancy and/or assisted reproductive technology.