Purpose To investigate the correlation of PNCK and EGFR expression with EGFR gene mutation status in non-small cell lung cancer(NSCLC).Methods Immunohisto-chemical staining was used to detect the expression of PNCK and EGFR in 256 cases of invasive lung carcinoma,42 cases of inva-sive precancerous lesions and 40 cases of paracancerous lung tis-sues,EGFR gene amplification and mutation were detected by next-generation sequencing.The relationship between the ex-pression of PNCK and EGFR protein and the clinicopathological features of NSCLC,the mutation status of EGFR gene and the correlation between them were analyzed.GEPIA database was used to analyze the correlation between PNCK and EGFR gene expression in lung adenocarcinoma and lung squamous cell carci-noma,and Kaplan-Meier Plotter database was used to analyze the effect of PNCK on the prognosis of lung cancer.Results Expression of PNCK and EGFR in lung invasive carcinoma(53.5％,137/256;47.7％,122/256)was higher than those of pre-invasive lesions(47.6％,20/42;31.0％,13/42)and paracancer tissue(22.5％,9/40;7.5％,3/40),pair-to-pair comparison among the three groups had statistical significance(P＜0.05).The high expression of PNCK protein was correla-ted with the degree of cancer differentiation,TNM stage,lymph node metastasis and distant metastasis(P＜0.05).The high expression of EGFR protein was correlated with TNM stage,lymph node metastasis and distant metastasis(P＜0.05).There was a negative correlation between PNCK and EGFR pro-tein expression in NSCLC tissues(r=-0.208,P＜0.05).The expression of EGFR protein in NSCLC with EGFR gene mu-tation and amplification was higher than that of wild type(P＜0.05).There was no correlation between PNCK protein expres-sion and EGFR gene mutation and amplification(P＞0.05).Bioinformatics database showed that PNCK and EGFR gene ex-pression were positively correlated in lung adenocarcinoma and lung squamous cell carcinoma(r=0.15,P＜0.05;r=0.30,P＜0.05),PNCK high expression group had a shorter overall survival rate in lung cancer(HR:1.28,95％CI:1.09-1.51,P=0.003 1).Conclusion The high expression of PNCK is associated with the occurrence,development,and poor prognosis of NSCLC,and negatively correlated with EGFR pro-tein expression.It is expected to become a potential therapeutic target for EGFR mutations in NSCLC.

Objectives:To analyze the impact of target lesion calcification evaluated by coronary angiography on immediate procedure success rate and long-term clinical outcome in patients underwent percutaneous coronary intervention(PCI). Methods:Consecutive patients received angiographic calcification evaluation and underwent PCI in Fuwai Hospital,Chinese Academy of Medical Sciences from January 2017 to December 2018 were prospectively enrolled in this study.Patients are divided into 4 groups,including non-calcification group(n=14 387),mild calcification group(n=8 231),moderate calcification group(n=3 208)and severe calcification group(n=886).The primary endpoint was immediate post-PCI procedure success rate,which was defined as residual stenosis<50%without major operational complications(including coronary artery perforation,coronary artery dissection,intraoperative stent thrombosis)and failure to cross the lesion.The long-term clinical endpoint was the 3-year major adverse cardiovascular events(MACE),which included composite endpoint events such as all cause death,myocardial infarction(MI),and revascularization. Results:The severe calcification group had a lower success rate of interventional therapy(80.59%vs.94.23%vs.94.29%vs.91.49%),a higher incidence of residual stenosis≥50%(17.72%vs.5.03%vs.4.84%vs.7.29%),a higher incidence of coronary artery dissection(1.81%vs.0.66%vs.0.78%vs.1.25%),and a higher incidence of coronary artery perforation(2.14%vs.0.51%vs.0.38%vs.0.69%),and a higher incidence of failure to cross the lesion(0.23%vs.0.12%vs.0.04%vs.0.16%)compared to the non-calcification group,mild calcification group,and moderate calcification group(all P<0.05).Multivariate Cox regression analysis showed that patients with severe calcification had a higher risk of MACE(12.98%vs.9.35%,HR=1.21,95%CI:1.00-1.47,P=0.046),all-cause mortality(4.29%vs.1.80%,HR=1.55,95%CI:1.10-2.18,P=0.013),and MI(2.14%vs.0.97%,HR=1.97,95%CI:1.21-3.20,P=0.006)compared to patients without calcification. Conclusions:Patients with angiographic-detected severe calcification after PCI treatment face higher risk of PCI procedure failure,MACE,all-cause death,and MI compared to patients without,with mild,and moderate calcification.

In order to solve the problems of quality control and traceability of medical test lung for meeting the calibration conditions of JJF 1234-2018 Calibration Specification for Ventilators, the calibration device and method are researched for compliance and airway resistance of medical test lung in this paper. A calibration device for medical test lung is designed using constant volume active piston technology to simulate human breathing. Through comparison experiment, the deviation between this device and the similar foreign device can be found. The deviation is lower than 0.4% for lung compliance and lower than 0.7% for airway resistance. The calibration of lung compliance and airway resistance can be completed by this device. This device has a clear and complete traceability path to ensure quality control from the source. The calibration of ventilator is improved. This paper provides a reference for related metrology departments and medical institutions to study on quality inspection of respiratory medical instruments.
Humans ; Calibration ; Ventilators, Mechanical ; Respiration ; Quality Control ; Lung

Objective:To evaluate the safety and feasibility of neoadjuvant chemotherapy (NACT) combined with radical surgery for elderly patients with locally advanced gastric cancer (LAGC).Methods:One hundred and fourty eight patients with LAGC after NACT and gastrectomy between 2012 and 2020 were retrospectively reviewed. They were divided into two groups: (1) <65 years old (111 cases) and (2) ≥65 years old (37 cases) and their clinicopathological and prognostic data were compared.Results:There was no significant difference between the two groups in the incidence of hematological complications such as anemia ( χ2=0.235, P=0.628), leukopenia ( χ2=0.613, P=0.434), neutropenia ( χ2=0.011, P=0.918) and thrombocytopenia ( χ2=0.253, P=0.615) and non-hematological complications such as nausea ( χ2=0.092, P=0.762), vomiting ( χ2=0.166, P=0.683), diarrhea ( χ2=0.015, P=0.902) and mucositis ( χ2=0.199, P=0.766) due to NACT. There were no statistical differences between the older patients and the younger in operation duration ( t=0.270, P=0.604), intraoperative bleeding ( t=1.140, P=0.250) and R 0 resection rate ( χ2=0.105, P=0.750). The incidence of postoperative complications was 25.2% and 37.8% in the younger patients and the olders ( χ2=2.172, P=0.141). Pleural effusion ( χ2=7.007, P=0.008) and pulmonary infection ( χ2=10.204, P=0.001) was significantly higher in the older patients than in the youngers. The 3-year progression-free survival rate ( t=0.494, P=0.482) and 3-year overall survival rate ( t=0.013, P=0.908) were comparable between the two groups. Conclusions:NACT combined with radical surgery is safe and effective in elderly patients with LAGC, except for higher perioperative pulmonary-related complications.

Cell migration is defined as the directional movement of cells toward a specific chemical concentration gradient, which plays a crucial role in embryo development, wound healing and tumor metastasis. However, current research methods showed low flux and are only suitable for single-factor assessment, and it was difficult to comprehensively consider the effects of other parameters such as different concentration gradients on cell migration behavior. In this paper, a four-channel microfluidic chip was designed. Its characteristics were as follows: it relied on laminar flow and diffusion mechanisms to establish and maintain a concentration gradient; it was suitable for observation of cell migration in different concentration gradient environment under a single microscope field; four cell isolation zones (20 μm width) were integrated into the microfluidic device to calibrate the initial cell position, which ensured the accuracy of the experimental results. In particular, we used COMSOL Multiphysics software to simulate the structure of the chip, which demonstrated the necessity of designing S-shaped microchannel and horizontal pressure balance channel to maintain concentration gradient. Finally, neutrophils were incubated with advanced glycation end products (AGEs, 0, 0.2, 0.5, 1.0 μmol·L ^-1), which were closely related to diabetes mellitus and its complications. The migration behavior of incubated neutrophils was studied in the 100 nmol·L ^-1 of chemokine (N-formylmethionyl-leucyl-phenyl-alanine) concentration gradient. The results prove the reliability and practicability of the microfluidic chip.
Cell Movement ; Chemotaxis ; Equipment Design ; Lab-On-A-Chip Devices ; Microfluidic Analytical Techniques ; Microfluidics ; Neutrophils ; Reproducibility of Results

OBJECTIVE To establis h the method for the simultaneous determination of six iridoids （loganic acid ，loganin， sweroside，dipsanoside B ，dipsanoside A ，sylvestroside Ⅰ）and one triterpene saponin （asperosaponin Ⅵ）in Dipsacus asper . METHODS High performance liquid chromatography （HPLC） method was adopted. The determination was performed on Symmetry® C18 column with mobile phase consisted of acetonitrile- 0.1％ phosphoric acid solution （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelengths were set at 212 nm（asperosaponin Ⅵ）and 237 nm（dipsanoside B ，dipsanoside A ， sweroside，loganic acid ，sylvestroside Ⅰ，loganin）. The column temperature was set at 30 ℃，and sample size was 20 μL. RESULTS The linear range of loganic acid ， loganin， sweroside， sylvestroside Ⅰ ， dipsanoside B ， dipsanoside A and asperosaponin Ⅵ were 399.24-931.56，50.30-150.90，48.24-168.84，27.00-70.20，12.93-38.80，40.64-121.92，42.08-147.28 µg/mL （all r＞0.999 0）. RSDs of precision ，reproducibility and stability tests （24 h）were all less than 2％. Average recoveries were 104.43％（RSD＝0.63％，n＝6），101.74％（RSD＝1.11％，n＝6），100.76％（RSD＝1.06％，n＝6），98.00％（RSD＝1.58％，n＝6）， 99.03％（RSD＝2.31％，n＝6），102.93％（RSD＝2.26％，n＝6），102.31％（RSD＝1.00％，n＝6），respectively，The contents were 142.5-280.6，5.5-49.0，28.0-112.9，7.2-35.8，4.4-16.9，17.2-79.3，0.8-54.5 mg/g，respectively. CONCLUSIONS Established method is accurate and reliable ，and can be used for the content determination of 7 components in D. asper .

Objective:To investigate the auxiliary diagnostic value of serum midkine (MK) and thyroid stimulating hormone (TSH) for differentiated thyroid cancer (DTC).Methods:Seventy-one postoperative DTC patients (DTC group) treated with 131I were selected, and 143 patients with benign thyroid lesions (benign thyroid disease group) treated with surgery in Center Hospital of Xiaogan from March 2019 to December 2020 at the same period were also selected. Clinical data such as liver and kidney function indexes, positive rate of anti thyroglobulin antibodies (TGAb) and positive rate of thyroid peroxidase antibody (TPOAb) were collected before treatment, and their fasting blood samples were collected before treatment. Fully automated electrochemiluminescence immunoassay was used to measure free thyroxine (FT 4), free triiodothyronine (FT 3), TSH levels in patients′ serum. The serum MK levels were measured by enzyme-linked immunosorbent assay (ELISA). Binary Logistic regression model was used to screen for independent risk factors for the development of DTC. A receiver operating characteristic curve (ROC) was drawn to evaluate the efficacy of MK, TSH and MK combined with TSH, in aiding the diagnosis of DTC and its staging. Results:Serum TSH and MK levels in DTC group were higher than those in benign thyroid disease group: (3.55 ± 0.61) mU/L vs. (2.97 ± 0.46) mU/L, (394.25 ± 63.36) ng/L vs. (311.45 ± 42.66) ng/L, and the differences were statistically significant ( P<0.05). Elevated serum TSH and MK levels were independent risk factors for DTC. When MK combined with TSH was used to diagnose DTC, the area under the curve (AUC), sensitivity and specificity were higher than those of MK and TSH alone (0.925 vs. 0.859 and 0.783, 83.10% vs. 78.87% and 73.24%, 89.51% vs. 85.31% and 79.02%), and the differences were statistically significant ( P<0.05). Serum TSH and MK levels in stage Ⅲ and Ⅳ patients in DTC group were higher than those in stage Ⅰ and Ⅱ patients: (3.79 ± 0.65) mU/L vs. (3.42 ± 0.56) mU/L, (427.88 ± 52.73) ng/L vs. (311.45 ± 42.66) ng/L, and the differences were statistically significant ( P<0.05). The AUC, sensitivity and specificity of MK combined with TSH in the diagnosis of different stages of DTC were higher than those of MK and TSH alone (0.822 vs. 0.657 and 0.666, 73.90% vs. 56.52% and 56.52%, 83.33% vs. 77.08% and 79.17%), and the differences were statistically significant ( P<0.05). Conclusions:Serum TSH and MK levels are independent risk factors for the occurrence of DTC in patients, and the combination of them has certain auxiliary diagnostic value for the identification and staging of DTC.

Objective:To evaluate the clinical outcomes of additional surgery after non-curative endoscopic submucosal dissection (ESD) for early gastric cancer.Methods:Sixty-nine patients with early gastric cancer who underwent ESD and were diagnosed as having non-curative resection by postoperative pathology at Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from January 2014 to December 2020 were included in the retrospective observation. Patients were divided into the additional surgery group ( n=12) and the follow-up group ( n=57). The differences in clinical and pathological data of the two groups, the surgical outcomes of the additional surgery group, three-year recurrence-free survival and tumor-specific survival of the two groups, and the independent risk factors affecting three-year recurrence-free survival in the follow-up group were analyzed. Results:Compared with the follow-up group, the rates of submucosal infiltration [66.7% (8/12) VS 21.1% (12/57), χ 2=7.927, P=0.005], vascular invasion [33.3% (4/12) VS 1.8% (1/57), P=0.003] and nerve invasion [16.7% (2/12) VS 0.0% (0/57), P=0.028] in the additional surgery group were significantly higher. In the additional surgery group, the interval between the additional surgery and ESD was 18.5 d (7-55 d), the surgical time was 286.4±85.9 min, and the number of dissected lymph nodes was 25.6±7.4. Four patients (33.3%) had residual tumor. Postoperative complications occurred in 4 patients (33.3%) (all were discharged after conservative treatment), and there was no perioperative death. One patient developed liver metastases 17 months after the surgery, and died 22 months after surgery due to liver metastases. One patient died 22 months after surgery due to non-tumor causes. The three-year recurrence-free survival and three-year tumor-specific survival in additional surgery group were 91.7% (11/12) and 91.7% (11/12), respectively, and those in the follow-up group were 87.7% (50/57) and 100.0% (57/57), respectively. Multivariate Cox regression analysis showed that tumor size ≥2 cm was an independent risk factor for three-year recurrence-free survival in the follow-up group ( P=0.037, HR=15.595, 95% CI: 1.181-205.952). Conclusion:Additional surgery and close follow-up are safe and feasible therapeutic strategies for early gastric cancer patients who underwent non-curative ESD. Clinicians should make reasonable choice based on the pathological results, patients' physical condition and surgery intention. But for patients with primary tumor size ≥2 cm, additional surgery is recommended.

Objective : To explore the role and molecular mechanism of Supervillin in zebrafish embryo development. Methods : Clustal Omega and DNAman software were used to analyze and compare the homology of amino acid sequences of Svil and SVIL in zebrafish and human. The expression pattern of Svila (subtype of Svil protein in zebrafish) during early embryonic development was analyzed by RT⁃PCR and in situ hybridization. Svila expression in zebrafish was inhibited by injecting morphinos ( MO) , and morphological changes of embryos were observed. The expression and nuclear localization of β ⁃catenin protein were detected by Western blot , and the expression of Wnt/β⁃catenin target gene was detected by RT⁃PCR. Results : During the early embryonic development of zebrafish , Svila was maternally expressed and showed an upward trend with the development process. The expression of Svila was reduced by MO , and the development of zebrafish embryos was distorted and the body axis was bent , which might be related to the blocked movement of concentrated extension of embryos. Further studies showed that Svila expression affected β⁃catenin nuclear transport and Wnt/β⁃catenin signaling pathway activation. Conclusion Svila regulates the concentrated extension movement of zebrafish embryos by activating the Wnt/β⁃catenin signaling pathway.

Objective:To observe the lipogenic or osteogenic differentiation of decellularized adipose matrix (DAM) in the periosteal microenvironment.Methods:From June 2020 to December 2020, adipose tissue obtained by human liposuction was prepared as DAM at the National Institute of Plastic Surgery. Six male SD rats of 4 to 6 weeks were selected and implanted into the subperiosteum of the rat parietal bone according to the same initial volume. After 12 weeks, adipogenesis and osteogenic differentiation of DAM were observed by gross specimens, histological staining, and immunofluorescence staining. Label-free quantitative protein mass spectrometry was used for detection of osteogenic-associated proteins in DAM.Results:Vascularization around the DAM was evident. Adipogenesis and angiogenesis were observed in DAM by H&E and Masson staining, while OCN immunofluorescence staining confirmed osteogenic differentiation of DAM. The osteogenic differentiation related proteins were screened by mass spectrometry.Conclusions:In the microenvironment of periosteum, DAM mainly differentiates towards adipose tissue, but a few of them havs the potential to differentiate towards osteogenesis, which might be related to some of the osteogenesis-related proteins contained in DAM.