Purpose The study aimed to analyze the relationship between MET gene variants and clinicopathologi-cal features in patients with non-small cell lung cancer(NSCLC).Methods Next-generation sequencing technology was used to detect MET gene variants in NSCLC specimens.The association between MET gene variant status and clini-copathological features was then analyzed.Results Among 1 633 cases of NSCLC,the overall MET mutation rate was 4.53％(74/1 633).Variants were mainly observed in male patients,never-smokers,those older than 60 years,ade-nocarcinoma histology,and patients with TNM stage Ⅲ+Ⅳ disease(P＜0.05).MET gene variant status showed no significant assocication with patient age,sex,smoking history,or pathological subtype(P＞0.05),but was statistical-ly correlated with clinical stage and presence of distant metastasis(P＜0.05).The two major variant types were MET exon 14 skipping and MET amplification,which together accounted for 71.62％of all variants.In addition,MET am-plification was positively correlated with EGFR(P=0.003,rs=0.340)and TP53 mutations(P=0.002,rs=0.362),but showed no correlation with KRAS or ALK gene mutations.In contrast,MET exon 14 skipping was nega-tively correlated with EGFR gene mutations(P＜0.001,rs=-0.409),and showed no significant correlation with KRAS,ALK,or TP53 mutations.Conclusion Different types of MET gene variants(amplification,exon 14 skip-ping,fusion,and others)are significantly associated with clinical advanced clinical stage and distant metastasis in NSCLC,but are independent of patient age,sex,smoking history,and pathological subtype.MET amplification fre-quently co-occur with EGFR and TP53 co-mutations.

OBJECTIVE: To explore the clinical phenotype and genetic characteristics of a patient with Progressive pseudorheumatoid dysplasia (PPRD) due to compound heterozygous variants of CCN6 gene. METHODS: A patient who was admitted to Qilu Hospital of Shandong University due to "bilateral finger joint deformity, bilateral hip and knee joint movement limitation for 19 years" was selected as the study subject. Clinical data of the patient were retrospectively collected. Peripheral blood samples were collected from the patient and her parents and subjected to whole exome sequencing (WES). Long-read sequencing (LRS) and Sanger sequencing were used to verify the candidate variants. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the pathogenicity of candidate variants was classified. This study was approved by the Medical Ethics Committee of Qilu Hospital of Shandong University (Ethics No.: KYLL-202502 061). RESULTS: The patient, a 23-year-old female, presented with progressive polyarticular deformity, limited movement and abnormal growth and development since childhood. She was initially misdiagnosed as Ankylosing spondylitis and had poor response to sulphasalazine and etoricoxib treatment. WES revealed that she has harbored two heterozygous variants of the CCN6 gene (NM_198239.2), namely c.348C>A and c.676G>C. LRS confirmed that the two variants are located on two homologous chromosomes and constitute compound heterozygous variants. Based on the ACMG guidelines, both variants were rated as pathogenic (PVS1+PM2_Supporting+PM3; PM1+PM2_Supporting+PM3_Supporting+PM5+PP3_Strong). The c.676G>C variant has not been recorded by the HGMD and ClinVar databases. CONCLUSION The c.348C>A and c.676G>C compound heterozygous variants of the CCN6 gene probably underlay the pathogenesis of PPRD in this patient. Above finding has enriched the mutational spectrum of PPRD and provided a basis for the clinical diagnosis and genetic counseling.
Humans ; Female ; CCN Intercellular Signaling Proteins/genetics* ; Phenotype ; Heterozygote ; Young Adult ; Mutation ; Exome Sequencing ; Joint Diseases/congenital*

Objective To explore the regulatory mechanisms underlying the increased proportion of CD4+Foxp3+regulatory T(Treg)cells in late-stage diabetes mellitus(DM)with poorly-controlled blood glucose,and to identify new approaches and therapeutic targets for the prevention and treatment of secondary infections in the late stage of DM.Methods Wild-type C57BL/6 mice aged 6 to 8 weeks were randomly assigned to two groups,the experimental and the control groups(n=5 per group).Mice in the experimental group were injected with streptozotocin(STZ)to induce the mouse model of type 1 diabetes mellitus(T1D),while those in the control group received injection of an an equal volume of 0.1 mol/L citrate buffer.In addition,wild-type C57BL/6 mice aged 6 to 8 weeks were fed with high-fat diet for 2 months and subsequently randomly assigned to two groups,the experimental and the control groups(n=3 per group).Mice in the experimental group were injected with low-dose STZ for multiple times to induce the mouse model of type 2 diabetes mellitus(T2D),while those in the control group received an equal volume of 0.1 mol/L citrate buffer.The spleen and peripheral lymph nodes of the mice were collected 2 weeks after the stable onset of diabetes,and T cell immune responses were examined by flow cytometry.Naive T cells isolated by immunomagnetic beads were cultured to investigate the mechanisms by which high glucose regulates T cell differentiation and function.The frequency of Treg cells and effector T(Teff)cells,the expression levels of Ki67,a cell proliferation marker,cell apoptosis rate,and intracellular reactive oxygen species(ROS)levels in the mouse tissue single cell suspension and T cell culture samples were assessed by multicolor flow cytometry.Results Late-stage T1D and T2D mice with poorly-managed blood glucose exhibited increased peripheral CD4+Foxp3+Treg frequencies(P<0.05).In these diabetic mice with poorly-managed blood glucose,the expression of Ki67 in Treg cells was significantly upregulated(P<0.05),while the apoptosis of non-Treg cells(Foxp3-non-Treg cells)increased markedly(P<0.05).Under high-glucose treatment conditions,the ROS levels in Teff cells increased significantly,and the cell apoptosis also increased significantly.High-glucose treatment induced the activation of transforming growth factor-β(TGF-β)and promoted the differentiation of Treg cells,whereas blocking the TGF-β signaling pathway or neutralizing ROS completely inhibited high glucose-induced Treg differentiation(P<0.01).Conclusion Sustained hyperglycemic internal environment in poorly-controlled diabetic mice causes high level of ROS production in Teff cells by inducing oxidative stress,which leads to increased apoptosis of Teff cells,promotes the differentiation of Treg cells by activating TGF-β,and ultimately leads to exacerbated immunosuppressive environment in the late stages of DM.Inhibiting the high level of ROS in late-stage diabetic patients may be conducive to mitigating Teff apoptosis and increasing the frequencies of Treg cells,and may offer new perspectives for improving hyperglycemia-induced immunosuppression and secondary infections in the late stage of DM.

Objective:To explore the clinical phenotype and genetic characteristics of a patient with Progressive pseudorheumatoid dysplasia (PPRD) due to compound heterozygous variants of CCN6 gene. Methods:A patient who was admitted to Qilu Hospital of Shandong University due to " bilateral finger joint deformity, bilateral hip and knee joint movement limitation for 19 years" was selected as the study subject. Clinical data of the patient were retrospectively collected. Peripheral blood samples were collected from the patient and her parents and subjected to whole exome sequencing (WES). Long-read sequencing (LRS) and Sanger sequencing were used to verify the candidate variants. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the pathogenicity of candidate variants was classified. This study was approved by the Medical Ethics Committee of Qilu Hospital of Shandong University (Ethics No.: KYLL-202502 061).Results:The patient, a 23-year-old female, presented with progressive polyarticular deformity, limited movement and abnormal growth and development since childhood. She was initially misdiagnosed as Ankylosing spondylitis and had poor response to sulphasalazine and etoricoxib treatment. WES revealed that she has harbored two heterozygous variants of the CCN6 gene (NM_198239.2), namely c. 348C>A and c. 676G>C. LRS confirmed that the two variants are located on two homologous chromosomes and constitute compound heterozygous variants. Based on the ACMG guidelines, both variants were rated as pathogenic (PVS1+ PM2_Supporting+ PM3; PM1+ PM2_Supporting+ PM3_Supporting+ PM5+ PP3_Strong). The c. 676G>C variant has not been recorded by the HGMD and ClinVar databases. Conclusion:The c. 348C>A and c. 676G>C compound heterozygous variants of the CCN6 gene probably underlay the pathogenesis of PPRD in this patient. Above finding has enriched the mutational spectrum of PPRD and provided a basis for the clinical diagnosis and genetic counseling.

Objective To investigate the effect of extracorporeal diaphragmatic pacing(EDP)on wea-ning patients undergoing mechanical ventilation after acute Stanford type A aortic dissection(ATAAD)sur-gery.Methods A total of 80 patients who underwent ATAAD surgery and required mechanical ventilation in the cardiac intensive care unit(CCU)of our hospital from October 2023 to October 2024 were selected as the study subjects.They were randomly divided into a control group and an intervention group using a random number table method,with 40 patients in each group.Both groups of patients received conventional clinical drug therapy,mechanical ventilation support,and rehabilitation exercises after surgery.The intervention group additionally underwent EDP therapy on this basis.The clinical data,mechanical ventilation time,weaning out-comes,oxygenation index,tidal volume,and changes in diaphragm function parameters were compared be-tween two groups of patients.Results Compared with the control group,the intervention group had a shorter mechanical ventilation time[77.00(59.00,133.63)h]vs.54.50(49.00,105.25)h],offline success rate(72.5％vs.90.0％),oxygenation index[202.90(178.63,240.05)mmHg vs.236.50(196.00,293.75)mm-Hg],higher tidal volume[(496.00±111.95)mL vs.(563.23±133.98)mL],with a statistically significant difference(P＜0.05).Compared with the control group,diaphragmatic excursion(DE)of both sides[11.90(8.50,16.50)mm vs.15.30(12.20,19.80)mm;12.60(10.80,14.90)mm vs.14.70(11.43,19.20)mm]and diaphragmatic thickening fraction(DTF)of left and right side[21.75％(17.53％,27.13％)vs.26.30％(21.10％,32.40％);24.00％(17.68％,29.35％)vs.28.30％(22.43％,33.40％)]of the intervention group were higher,and the differences were statistically significant(P＜0.05).Conclusion EDP can enhance the function of the diaphragm in patients,shorten the duration of mechanical ventilation,improve the success rate of wea-ning,and improve the weaning outcomes of mechanically ventilated patients after ATAAD surgery.

Purpose The study aimed to analyze the relationship between MET gene variants and clinicopathologi-cal features in patients with non-small cell lung cancer(NSCLC).Methods Next-generation sequencing technology was used to detect MET gene variants in NSCLC specimens.The association between MET gene variant status and clini-copathological features was then analyzed.Results Among 1 633 cases of NSCLC,the overall MET mutation rate was 4.53％(74/1 633).Variants were mainly observed in male patients,never-smokers,those older than 60 years,ade-nocarcinoma histology,and patients with TNM stage Ⅲ+Ⅳ disease(P＜0.05).MET gene variant status showed no significant assocication with patient age,sex,smoking history,or pathological subtype(P＞0.05),but was statistical-ly correlated with clinical stage and presence of distant metastasis(P＜0.05).The two major variant types were MET exon 14 skipping and MET amplification,which together accounted for 71.62％of all variants.In addition,MET am-plification was positively correlated with EGFR(P=0.003,rs=0.340)and TP53 mutations(P=0.002,rs=0.362),but showed no correlation with KRAS or ALK gene mutations.In contrast,MET exon 14 skipping was nega-tively correlated with EGFR gene mutations(P＜0.001,rs=-0.409),and showed no significant correlation with KRAS,ALK,or TP53 mutations.Conclusion Different types of MET gene variants(amplification,exon 14 skip-ping,fusion,and others)are significantly associated with clinical advanced clinical stage and distant metastasis in NSCLC,but are independent of patient age,sex,smoking history,and pathological subtype.MET amplification fre-quently co-occur with EGFR and TP53 co-mutations.

Objective:To explore the clinical phenotype and genetic characteristics of a patient with Progressive pseudorheumatoid dysplasia (PPRD) due to compound heterozygous variants of CCN6 gene. Methods:A patient who was admitted to Qilu Hospital of Shandong University due to " bilateral finger joint deformity, bilateral hip and knee joint movement limitation for 19 years" was selected as the study subject. Clinical data of the patient were retrospectively collected. Peripheral blood samples were collected from the patient and her parents and subjected to whole exome sequencing (WES). Long-read sequencing (LRS) and Sanger sequencing were used to verify the candidate variants. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the pathogenicity of candidate variants was classified. This study was approved by the Medical Ethics Committee of Qilu Hospital of Shandong University (Ethics No.: KYLL-202502 061).Results:The patient, a 23-year-old female, presented with progressive polyarticular deformity, limited movement and abnormal growth and development since childhood. She was initially misdiagnosed as Ankylosing spondylitis and had poor response to sulphasalazine and etoricoxib treatment. WES revealed that she has harbored two heterozygous variants of the CCN6 gene (NM_198239.2), namely c. 348C>A and c. 676G>C. LRS confirmed that the two variants are located on two homologous chromosomes and constitute compound heterozygous variants. Based on the ACMG guidelines, both variants were rated as pathogenic (PVS1+ PM2_Supporting+ PM3; PM1+ PM2_Supporting+ PM3_Supporting+ PM5+ PP3_Strong). The c. 676G>C variant has not been recorded by the HGMD and ClinVar databases. Conclusion:The c. 348C>A and c. 676G>C compound heterozygous variants of the CCN6 gene probably underlay the pathogenesis of PPRD in this patient. Above finding has enriched the mutational spectrum of PPRD and provided a basis for the clinical diagnosis and genetic counseling.

Objective To investigate the protective and treatment role of ulinastatin(UTI)on con-trast-induced acute kidney injury(CIAKI)in the elderly with coronary heart disease(CHD)and chronic kidney diseases(CKD).Methods A total of 321 elderly CHD inpatients complicated with CKD undergoing coronary angiography admitted in the First Medical Center of Chinese PLA Gen-eral Hospital from November 2021 to November 2022 were enrolled consecutively and then divid-ed into UTI group(n=161)and hydration group(n=160).Their cardiac and renal function pa-rameters were collected and analyzed before and 2 d after intervention.The changes in above pa-rameters and incidence of CIAK were observed and compared between the two groups.Results In 2 d after intervention,the UTI group had significantly lower Scr,urea,CysC,homocysteine and NT-proBNP,but higher eGFR than the hydration group(P＜0.01).There were 62 patients(62/321,19.3％)developing CIAKI,including 17 from the UTI group and 45 from the hydration group,and statistical difference was observed in the incidence(10.6％vs 28.1％,P＜0.01).For the patients with comorbidities of hypertension,diabetes,hyperlipidemia and hyperuricemia,the incidence of CIAKI was obviously lower in the UTI group than the hydration group(P＜0.01).Multivariate logistic regression analysis showed that UTI was an independent protective factor for occurrence of CIAKI(OR=0.348,95％CI:0.180-0.673,P=0.001).Conclusion UTI can im-prove renal function and reduce the risk of CIAKI in elderly CHD patients with CKD.

The corneal epithelium, an essential refractive interface, plays an integral role in the corneal healing after corneal refractive surgery. All existing corneal refractive surgeries entail a degree of corneal epithelial remodeling; however, excessive epithelial remodeling precipitates adverse outcomes on the refractive correction efficacy of such surgeries. This review summarizes the application of corneal epithelial remodeling in the corneal refractive surgery, and more comprehensively investigates the influencing factors of perioperative epithelial remodeling after corneal refractive surgery, with a view to augmenting the safety, efficacy, predictability, and stability of corneal refractive surgical outcomes.

ObjectiveTo study the effect and mechanism of Linggui Zhugantang in treating chronic bronchitis （CB） induced by exposure to cigarette smoke combined with tracheal instillation of lipopolysaccharide （LPS）. MethodSixty SPF-grade SD rats were randomly divided into normal， model， dexamethasone （1 mg·kg^-1）， and high-， medium-， and low-dose （30.06， 15.03， 7.515 g·kg^-1， respectively） Linggui Zhugantang groups by the body weight stratification method， with 10 rats in each group. Each group was administrated with 200 μL LPS （1 g·L^-1） by tracheal instillation on days 1 and 14， respectively， while the normal group was administrated with an equal volume of normal saline. Except the normal group， the other groups were exposed to cigarette smoke on days 2-13 and 15-30 （10 cigarettes/time/30 min， twice/day） for the modeling of CB. The rats were administrated with corresponding drugs by gavage for 30 consecutive days from day 2 of modeling， and the mental status， behavior， and body weights of the rats were observed and measured. The wet/dry mass ratio （W/D） of the left lung was measured 30 days after modeling. Hematoxylin-eosin staining was employed to observe the pathological changes in the lung and bronchial tissues. The bronchial mucus secretion and goblet cell proliferation were observed by Alcian blue-periodic acid Schiff （AB-PAS） staining. The levels of mucin 5AC （MUC5AC）， interleukin （IL）-13， IL-6， and tumor necrosis factor （TNF）-α in the serum were determined by enzyme-linked immunosorbent assay. The expression of phospholipase A2 （PLA2）， transient receptor potential vanilloid receptor 1 （TRPV1）， and transient receptor potential ankyrin 1 （TRPA1） in the lung tissue was quantitatively analyzed by immunohistochemistry and Western blot. ResultCompared with the normal group， the model group showcased abnormal mental status and behaviors， bloody secretion in the nose and mouth， the mortality rate of 40%， decreased body weight， severe lung bronchial structure damage， a large number of inflammatory mediators and inflammatory cell infiltration in the tube wall， hyperemia， edema， and fibroplasia， massive proliferation of goblet cells， excessive secretion and accumulation of mucus， stenosis and deformation of the lumen， and aggravation of pulmonary edema （P<0.01）. In addition， the model group had higher levels of MUC5AC， IL-13， IL-6， and TNF-α in the serum and higher expression of PLA2 in the lung tissue than the normal group （P<0.01）. Compared with the model group， the medication groups showed normal mental status and behaviors， reduced mortality rate， stable weight gain， reduced lung and bronchial injuries， decreased goblet cell proliferation and mucus secretion， and alleviated pulmonary edema （P<0.01）. Furthermore， Linggui Zhugantang lowered the levels of MUC5AC， IL-13， IL-6， and TNF-α in the serum and down-regulated the protein levels of PLA2， TRPV1， and TRPA1 in the lung tissue （P<0.01）. ConclusionLinggui Zhugantang can reduce the pulmonary inflammation and airway mucus hypersecretion in the rat model of chronic bronchitis. It may exert the effects of reducing inflammation and resolving phlegm by regulating the PLA2-TRPV1/TRPA1 pathway.