ObjectiveTo investigate the role and mechanism of Lck/Yes-related novel protein tyrosine kinase （Lyn） on lipopolysaccharide （LPS）-induced M1-type polarization of macrophage. MethodsThe LentiCRISPR-V2 plasmid was digested with the restriction endonuclease BSMBI-V2， and the digested DNA fragments were recovered. The digested plasmid was ligated with Lyn-sgRNA using T4 ligase to generate the Lenti-Lyn-gRNA lentivirus. THP-1 cells were infected with the Lenti-Lyn-gRNA lentivirus to obtain a stable cell line with Lyn knockout， and a monoclonal THP-1 cell line with complete Lyn knockout （Lyn^⁻/⁻） was established subsequently. Wild-type Lyn （Lyn^WT） and Lyn^⁻/⁻ THP-1 cells were induced with 100 ng/mL phorbol myristate acetate （PMA） for 48 h to differentiate into M0 macrophages， which were further polarized into M1 macrophages by stimulation with 100 ng/mL LPS for 24 h. Quantitative real-time polymerase chain reaction （qPCR） was performed to detect the expression of M0 macrophage markers， including integrin αM （CD11b）， macrophage antigen （CD68）， and monocyte differentiation antigen （CD14）. The expression of Lyn in M1 macrophages differentiated from wild-type THP-1 cells （Lyn^WT-M1） was measured by qPCR， and the ratio of phosphorylated Lyn to total Lyn （P-Lyn/Lyn） in Lyn^WT-M1 cells was determined by Western blot. In M1 macrophages differentiated from Lyn-knockout THP-1 cells （Lyn^⁻/⁻-M1）， qPCR was used to detect the mRNA expression of inducible nitric oxide synthase （iNOS）， interleukin-6 （IL-6）， and chemokine （C-X-C motif） ligand 10 （CXCL-10）. Western blot was conducted to assess the protein expression of iNOS， as well as the protein levels of molecules related to the Janus kinase 1 （JAK1）-signal transducer and activator of transcription 1 （STAT1）signaling pathway， including JAK1， phosphorylated JAK1 （P-JAK1）， STAT1， and phosphorylated STAT1 （P-STAT1）. Additionally， the expression of the M1 macrophage marker cluster of differentiation 80 （CD80） was analyzed by flow cytometry. ResultsThe Lyn^-/- monoclonal cell line was successfully constructed. The expression of CD11b was significantly elevated in Lyn^-/- M0 macrophages， and the differentiation of M1 macrophages was successful. Knockdown of Lyn inhibited mRNA expression of iNOS， IL⁃6， CXCL⁃10， protein expression of iNOS and CD80 expression in M1 macrophages （P<0.05）. Western blot assay showed that Lyn knockdown inhibited protein expression of JAK1 and P-STAT1 （P<0.01）. ConclusionAfter CRISPR/Cas9-mediated Lyn knockout， the expression levels of JAK1 and P-STAT1， the key molecules in the JAK/STAT signaling pathway of M1 macrophages， are significantly downregulated； concomitantly， the expression of M1 macrophage-specific secretory factors （iNOS， IL⁃6， CXCL⁃10 mRNA） and CD80 is also downregulated， which may be achieved via targeted regulation of the JAK1/P-STAT1-mediated JAK/STAT signaling pathway.

Hepatocellular carcinoma(HCC)is the most common type of liver cancer and a leading cause of cancer-related deaths worldwide.While early detection significantly improves prognosis,patients with alpha-fetoprotein-negative hepatocellular carcinoma(ANHCC)often face diagnostic challenges due to the lack of reliable biomarkers.This review systematically explores the potential of various circulating biomarkers in the early diagnosis of ANHCC,including AFP-L3,PIVKA-Ⅱ,lymphocyte-to-monocyte ratio,exosomes,circulating cell-free DNA(cfDNA),circulating tumor cells,osteopontin,paraoxonase 1,autoantibodies,and RNA-related biomarkers.The combined use of these markers,particularly AFP-L3 and PIVKA-Ⅱ,demonstrates enhanced diagnostic accuracy and specificity compared to single markers.Emerging evidence also highlights the diagnostic potential of exosomes,cfDNA,and RNA markers due to their non-invasive nature and high stability.Despite promising results,further large-scale,multicenter studies are needed to validate these findings,address challenges such as standardization of detection methods,and elucidate underlying mechanisms.These advances are anticipated to significantly improve early detection and personalized management of ANHCC.

Objective Cone beam computed tomograph(CBCT)was used to investigate the factors influencing the difficulty of extracting impacted mandibular third molar(IMTM).Methods The CBCT image data of 441 teeth of 260 patients who visited Affiliated Stomatological Hospital of Huzhou University from March 2023 to June 2024 were selected.Record the number of roots,impacted status,direction of impacted teeth,depth of impaction,and positional relationship with mandibular nerve canal of impacted teeth,and classify them according to different genders.Statistically analyze the differences between different genders.Results Among the 441 IMTM teeth,there were 82 single root,333 double roots,24 triple roots and 2 quadruple roots.Impacted status:179 inclined impaction,129 vertical impaction,128 horizontal impaction,5 inverted impaction.Direction of impacted teeth:133 mesioangular impaction,7 buccoangular impaction,29 linguoangular impaction,10 distoangular impaction.Depth of impaction:285 high impaction,124 medium impaction,32 low impaction.The positional relationship with mandibular nerve canal:there are 159 teeth with roots located on the lingual side of mandibular nerve canal,140 teeth with roots entering mandibular nerve canal,22 teeth with roots on the buccal side of mandibular nerve canal,117 teeth with roots above mandibular nerve canal,1 tooth with roots between them,and 2 special teeth.There were statistically significant differences in the number of roots,impacted status,and direction of impacted teeth among patients of different genders(P＜0.05),however there were no statistically significant differences in depth of impaction and positional relationship with mandibular nerve canal(P＞0.05).Conclusion There are differences in morphological characteristics of IMTM between different genders,which leads to different extraction difficulties.CBCT can precisely observe the morphological characteristics of IMTM from a three-dimensional perspective,providing guidance for the clinical formulation of extraction strategies for impacted teeth.

Objective Cone beam computed tomograph(CBCT)was used to investigate the factors influencing the difficulty of extracting impacted mandibular third molar(IMTM).Methods The CBCT image data of 441 teeth of 260 patients who visited Affiliated Stomatological Hospital of Huzhou University from March 2023 to June 2024 were selected.Record the number of roots,impacted status,direction of impacted teeth,depth of impaction,and positional relationship with mandibular nerve canal of impacted teeth,and classify them according to different genders.Statistically analyze the differences between different genders.Results Among the 441 IMTM teeth,there were 82 single root,333 double roots,24 triple roots and 2 quadruple roots.Impacted status:179 inclined impaction,129 vertical impaction,128 horizontal impaction,5 inverted impaction.Direction of impacted teeth:133 mesioangular impaction,7 buccoangular impaction,29 linguoangular impaction,10 distoangular impaction.Depth of impaction:285 high impaction,124 medium impaction,32 low impaction.The positional relationship with mandibular nerve canal:there are 159 teeth with roots located on the lingual side of mandibular nerve canal,140 teeth with roots entering mandibular nerve canal,22 teeth with roots on the buccal side of mandibular nerve canal,117 teeth with roots above mandibular nerve canal,1 tooth with roots between them,and 2 special teeth.There were statistically significant differences in the number of roots,impacted status,and direction of impacted teeth among patients of different genders(P＜0.05),however there were no statistically significant differences in depth of impaction and positional relationship with mandibular nerve canal(P＞0.05).Conclusion There are differences in morphological characteristics of IMTM between different genders,which leads to different extraction difficulties.CBCT can precisely observe the morphological characteristics of IMTM from a three-dimensional perspective,providing guidance for the clinical formulation of extraction strategies for impacted teeth.

Hepatocellular carcinoma(HCC)is the most common type of liver cancer and a leading cause of cancer-related deaths worldwide.While early detection significantly improves prognosis,patients with alpha-fetoprotein-negative hepatocellular carcinoma(ANHCC)often face diagnostic challenges due to the lack of reliable biomarkers.This review systematically explores the potential of various circulating biomarkers in the early diagnosis of ANHCC,including AFP-L3,PIVKA-Ⅱ,lymphocyte-to-monocyte ratio,exosomes,circulating cell-free DNA(cfDNA),circulating tumor cells,osteopontin,paraoxonase 1,autoantibodies,and RNA-related biomarkers.The combined use of these markers,particularly AFP-L3 and PIVKA-Ⅱ,demonstrates enhanced diagnostic accuracy and specificity compared to single markers.Emerging evidence also highlights the diagnostic potential of exosomes,cfDNA,and RNA markers due to their non-invasive nature and high stability.Despite promising results,further large-scale,multicenter studies are needed to validate these findings,address challenges such as standardization of detection methods,and elucidate underlying mechanisms.These advances are anticipated to significantly improve early detection and personalized management of ANHCC.

As the search for effective treatments for COVID-19 continues, the high mortality rate among critically ill patients in Intensive Care Units (ICU) presents a profound challenge. This study explores the potential benefits of traditional Chinese medicine (TCM) as a supplementary treatment for severe COVID-19. A total of 110 critically ill COVID-19 patients at the Intensive Care Unit (ICU) of Vulcan Hill Hospital between Feb., 2020, and April, 2020 (Wuhan, China) participated in this observational study. All patients received standard supportive care protocols, with a subset of 81 also receiving TCM as an adjunct treatment. Clinical characteristics during the treatment period and the clinical outcome of each patient were closely monitored and analysed. Our findings indicated that the TCM group exhibited a significantly lower mortality rate compared with the non-TCM group (16 of 81 vs 24 of 29; 0.3 vs 2.3 person/month). In the adjusted Cox proportional hazards models, TCM treatment was associated with improved survival odds (P < 0.001). Furthermore, the analysis also revealed that TCM treatment could partially mitigate inflammatory responses, as evidenced by the reduced levels of proinflammatory cytokines, and contribute to the recovery of multiple organic functions, thereby potentially increasing the survival rate of critically ill COVID-19 patients.
Humans ; COVID-19 ; Medicine, Chinese Traditional ; SARS-CoV-2 ; Critical Illness ; Treatment Outcome

【Objective】 To analyze the expression of lncRNA SNHG25 in prostate cancer and its significance, so as to explore the biomarkers and potential therapeutic targets for the diagnosis and prognosis of this disease. 【Methods】 Based on the TCGA database, differential, survival, and clinical correlation analyses of SNHG25 were performed.SNHG25 expression in prostate cancer was analyzed in the UALCAN database to determine its relationship with the clinical and pathological characteristics.The lncRNA-miRNA-mRNA correlation analysis was performed.The relevant ceRNA regulatory network was constructed.Prostate cancer samples were divided into high and low SNHG25 expression groups, and differential SNHG25 related genes were filtered and then enriched. 【Results】 SNHG25 expression was significantly upregulated in prostate cancer specimens compared to normal prostate specimens (P<0.001), and the progression-free survival of the low SNHG25 expression group was significantly longer than that of the high SNHG25 expression group (P<0.001).There were no significant differences in age, T-stage and N-stage between the two groups, and there was no significant correlation between the expression of SNHG25 and Gleason score (P>0.05).Regulatory networks of SNHG25/miR-330-3p/DLX1 and RPL22L1 were constructed. 【Conclusion】 SNHG25 is highly expressed in prostate cancer tissues and correlated with poor prognosis.SNHG25 expression does not significantly correlate with age, T-stage, N-stage, and Gleason score.SNHG25/miR-330-3p/DLX1 and RPL22L1 regulatory networks may play an important role in the development of prostate cancer.SNHG25 may become a biomarker and potential therapeutic target for prostate cancer.

Objective:To explore the diagnosis and clinical characteristics of congenital meningocele combined with type 1 neurofibromatosis (NF1).Methods:Clinical data of a child with congenital meningoencephalocele complicated with NF1 diagnosed and treated by the Hunan Provincial Brain Hospital on July 12, 2021 were collected, and relevant literature was reviewed.Results:Patient, female, 9 years old, with milk coffee stains on the skin of the trunk and limbs, mostly with a length greater than 1.5 cm; A three-dimensional CT scan of the skull showed a skull defect in the left temporal occipital region (approximately 7.5 cm×6.7 cm) and below the right posterior occipital tuberosity (approximately 3 cm×3 cm). The patient′s clinical whole exome sequencing detected one pathogenic mutation (NF1: p. Arg681Ter), and underwent lumbar cistern drainage, cyst resection, occipital cistern ostomy, and titanium mesh repair and shaping surgery. The pathological result showed proliferation of arachnoid and fibrous tissue, and good postoperative recovery.Conclusions:Congenital meningoencephalocele combined with NF1 is extremely rare, and early surgery has more advantages than disadvantages. The use of titanium mesh with sliding groove can alleviate the limitation on skull growth to a certain extent. NF1 has diverse clinical manifestations and different onset ages, and should be followed up by a multidisciplinary team for a long time.

Objective:To compare the clinical effects of ultrasound-guided intra-articular and coracohumeral ligament glucocorticoid injections versus ultrasound-guided intra-articular glucocorticoid injections alone on frozen shoulder in the elderly.Methods:A total of 44 patients with frozen shoulder admitted to the rehabilitation department of the Affiliated Hospital of Sichuan Nursing Vocational College from September 2019 to June 2020 were enrolled.They were randomly divided into the experimental group and the control group, with 22 patients in each group.The experimental group received two consecutive ultrasound-guided intra-articular injections and two coracohumeral ligament injections, while the control group received two consecutive ultrasound-guided intra-articular injections.The shoulder's passive range-of-motion(PROM), the 11-point numeric rating scale(NRS)as well as the Shoulder Pain and Disability Index(SPADI)were conducted before treatment, 1 week after the first treatment(1 week), 1 week after the second treatment(2 weeks)and 10 weeks after the second treatment(12 weeks).Results:There was no significant difference in baseline characteristics between the two groups(all P>0.05). Flexion, abduction, extension and external rotation increased whereas NRS and SPADI decreased after treatment, compared with pre-treatment in the experimental group( F=11.341, 20.965, 20.403, 32.470, 82.970 and 102.154, all P<0.05)and in the control group( F=7.012, 8.937, 23.265, 31.966, 52.500 and 41.356, all P<0.05). Compared with the control group, extension( t=0.365, 0.143 and 0.236, all P<0.05)and external rotation( t=0.205, 0.620 and 2.751, all P<0.05)increased at 1, 2 and 12 weeks after treatment.SPADI scores were lower in the experimental group than in the control group at 12 weeks after treatment( t=2.063, P=0.045). There was no significant difference in NRS, flexion or abduction between the two groups at any time point after treatment(all P>0.05). No serious adverse reactions occurred during treatment in either group. Conclusions:Both ultrasound-guided intra-articular injections plus coracohumeral ligament injections and ultrasound-guided intra-articular injections can effectively alleviate pain and improve function in elderly patients with frozen shoulder, but the former has better effects on extension.For patients with clear restricted extension and external rotation, ultrasound-guided intra-articular injections combined with coracohumeral ligament injections may achieve better clinical effects.

Perception ; Pupil ; Social Interaction