Objective:To improve the diagnosis and treatment of thyroid carcinoma in flying personnel by analyzing the clinical characteristics of thyroid carcinoma.Methods:The clinical and pathological data of flying personnel with thyroid carcinoma diagnosed and treated by Air Force Medical Center of PLA from January of 2010 to December of 2022 were retrospectively analyzed. Clinical characteristics such as age, flying hours, aircraft types, thyroid function at diagnosis, and pathological characteristics such as tumor site, tumor size, lymph node metastasis, calcification and sand and Ki-67 positive ratio were collected, and the clinical and pathological characteristics were statistically analyzed.Results:Thyroid nodule was found as the first diagnosis of thyroid carcinoma in 57 flying personnel. The age of onset was 22-58 years old, the median age was 37 years old, the flying hours was 4-18 000 h, and the median flying hours was 2 000 h. Thyroid carcinoma was detected in both thyroid glands, and the histological type was papillary thyroid carcinoma. Thyroid function was normal in most cases when thyroid carcinoma was detected. Lymph node metastasis of thyroid carcinoma was low positively correlated with tumor size ( r=0.304, P=0.021), and was low positively correlated with the positive proportion of Ki-67 ( r=0.360, P=0.006). Other clinical and pathological characteristics did not show clear correlation. With the extension of flying hours, the incidence of lymph node metastasis of thyroid carcinoma increased, and the difference was statistically significant ( χ2=6.32, P=0.012). There was no statistical difference among the clinical characteristics of flying personnel with different age and aircraft types ( P>0.05). Conclusions:The thyroid carcinoma is usually diagnosed without clinical symptoms and the thyroid function is basically normal. The thyroid ultrasound examination should be emphasized during physical examination, and further examination should be conducted for the flying personnel with thyroid nodules.

Objective:To improve the diagnosis and treatment of thyroid carcinoma in flying personnel by analyzing the clinical characteristics of thyroid carcinoma.Methods:The clinical and pathological data of flying personnel with thyroid carcinoma diagnosed and treated by Air Force Medical Center of PLA from January of 2010 to December of 2022 were retrospectively analyzed. Clinical characteristics such as age, flying hours, aircraft types, thyroid function at diagnosis, and pathological characteristics such as tumor site, tumor size, lymph node metastasis, calcification and sand and Ki-67 positive ratio were collected, and the clinical and pathological characteristics were statistically analyzed.Results:Thyroid nodule was found as the first diagnosis of thyroid carcinoma in 57 flying personnel. The age of onset was 22-58 years old, the median age was 37 years old, the flying hours was 4-18 000 h, and the median flying hours was 2 000 h. Thyroid carcinoma was detected in both thyroid glands, and the histological type was papillary thyroid carcinoma. Thyroid function was normal in most cases when thyroid carcinoma was detected. Lymph node metastasis of thyroid carcinoma was low positively correlated with tumor size ( r=0.304, P=0.021), and was low positively correlated with the positive proportion of Ki-67 ( r=0.360, P=0.006). Other clinical and pathological characteristics did not show clear correlation. With the extension of flying hours, the incidence of lymph node metastasis of thyroid carcinoma increased, and the difference was statistically significant ( χ2=6.32, P=0.012). There was no statistical difference among the clinical characteristics of flying personnel with different age and aircraft types ( P>0.05). Conclusions:The thyroid carcinoma is usually diagnosed without clinical symptoms and the thyroid function is basically normal. The thyroid ultrasound examination should be emphasized during physical examination, and further examination should be conducted for the flying personnel with thyroid nodules.

A 71-year-old male patient with advanced lung adenocarcinoma received pemetrexed (0.8 g, IV infusion on the first day) and carboplatin (500 mg, IV infusion on the first day) combined with pembrolizumab (200 mg, IV infusion on the second day) and 21 days was a cycle. Before the third cycle of treatment, the patient developed palpitations, irritability, increased appetite, and emaciation. Laboratory tests showed triiodothyronine (T 3) 2.88 nmol/L, thyroxine (T 4) 247.90 nmol/L, free triiodothyronine (FT 3) 10.57 pmol/L, free thyroxine (FT 4) 39.63 pmol/L, thyroid stimulating hormone (TSH) 0.014 mU/L, anti-thyroglobulin antibody (TGAb) 15.9 μg/L, thyroid peroxidase antibody (TPOAb) >1 300.0 kU/L. Immunerelated hyperthyroidism was considered, which may be related to pembrolizumab. The above-mentioned treatment was continued due to the patient′s condition, and thiamazole and metoprolol were given orally at the same time. One month later, laboratory tests showed T 3 2.50 nmol/L, T 4 153.40 nmol/L, FT 3 7.70 pmol/L, FT 4 33.61 pmol/L, TSH 0.007 mU/L, TGAb 15.7 μg/L and TPOAb >1 300.0 kU/L; 2 months later, laboratory tests showed T 3 1.84 nmol/L, T 4 81.20 nmol/L, FT 3 3.86 pmol/L, FT 4 11.56 pmol/L, TSH 1.979 mU/L, TGAb 15.7 μg/L, and TPOAb >1 300.0 kU/L. His symptoms of palpitation and irritability were alleviated.

A 71-year-old male patient with advanced lung adenocarcinoma received pemetrexed (0.8 g, IV infusion on the first day) and carboplatin (500 mg, IV infusion on the first day) combined with pembrolizumab (200 mg, IV infusion on the second day) and 21 days was a cycle. Before the third cycle of treatment, the patient developed palpitations, irritability, increased appetite, and emaciation. Laboratory tests showed triiodothyronine (T 3) 2.88 nmol/L, thyroxine (T 4) 247.90 nmol/L, free triiodothyronine (FT 3) 10.57 pmol/L, free thyroxine (FT 4) 39.63 pmol/L, thyroid stimulating hormone (TSH) 0.014 mU/L, anti-thyroglobulin antibody (TGAb) 15.9 μg/L, thyroid peroxidase antibody (TPOAb) >1 300.0 kU/L. Immunerelated hyperthyroidism was considered, which may be related to pembrolizumab. The above-mentioned treatment was continued due to the patient′s condition, and thiamazole and metoprolol were given orally at the same time. One month later, laboratory tests showed T 3 2.50 nmol/L, T 4 153.40 nmol/L, FT 3 7.70 pmol/L, FT 4 33.61 pmol/L, TSH 0.007 mU/L, TGAb 15.7 μg/L and TPOAb >1 300.0 kU/L; 2 months later, laboratory tests showed T 3 1.84 nmol/L, T 4 81.20 nmol/L, FT 3 3.86 pmol/L, FT 4 11.56 pmol/L, TSH 1.979 mU/L, TGAb 15.7 μg/L, and TPOAb >1 300.0 kU/L. His symptoms of palpitation and irritability were alleviated.

Objective: To investigate the characteristics and clinical significance of the immunomicroenvironment typing based on the expression of programmed death-ligand 1 (PD-L1) and the infiltration of CD8+ T cells in the stroma in patients with non-small cell lung cancer (NSCLC). Methods: Paraffin tissue specimens and relevant clinicopathological data of 74 NSCLC patients admitted to our hospital from January 2016 to July 2018 were collected.All patients received EGFR gene test, and none received radiotherapy, chemotherapy or targeted therapy. Immunohistochemistry was used to detect the expression of PD-L1 in tissues and the infiltration of CD8+T cells in interstitium, and the relationship between PD-L1, CD8+T cells, and the immune microenvironment typing based on both, and the pathological parameters and the survival of patients was analyzed. Results: PD-L1 expression in the primary tumor of NSCLC patients showed statistical differences in gender, pathological type, smoking history, EFGR gene mutation status ( P <0.05). The infiltration of CD8+ T lymphocytes in tumor microenvironment showed statistically significant differences in different TNM stage and lymph node metastasis ( P <0.05), PD-L1 expression was significantly correlated with EGFR mutation ( P =0.000), while CD8+T lymphocyte infiltration was not correlated with EGFR mutation ( P =0.605). The immunomicroenvironment of EGFR wild-type patients was mainly (CD8+ PD-L1+) (type I), and the mutants were mainly (CD8-PD-L1-) (type II) and (CD8+PD-L1-) (type IV). The distribution of immune microenvironmental typing in each group with different EGFR mutation, smoking history and pathological differentiation degree was significantly different ( P <0.05) and significantly correlated with EGFR mutation ( P <0.05). Follow-up showed that the patients with disease free survival, recurrence and metastasis and death were the most in type I, type II and type I, respectively. Conclusions: In this study, the distribution of tumor immunomicroenvironmental typing in NSCLC patients was mainly the highest in type I and the lowest in type Ⅲ, which was related to EGFR mutation, smoking history and pathological differentiation. Patients with EGFR mutations were mainly of type Ⅱand type Ⅳ, and were associated with low expression of PD-L1. ··

OBJECTIVEThe aim of this study was to establish a standardized protocol for detection of ALK protein expression and gene fusion in cytologic specimens.

METHODSLung adenocarcinoma cytologic specimens were collected from seven hospitals in Beijing city. A detection protocol for ALK protein expression and gene fusion was designed according to the results of comparative experiment. Ventana immunohistochemical (IHC) ALK(D5F3) detecting ALK protein expression was performed in 203 prepared formalin-fixed paraffin-embedded (FFPE) cell blocks. ALK gene fusion in 98 EGFR gene wild type cytologic specimens and in 4 bronchoalveolar lavage fluid (BL) samples was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). ALK gene fusion in the Ventana IHC ALK (D5F3) positive samples was further tested by fluorescence in situ hybridization (FISH). Six patients with ALK IHC-positive result were followed up to analyze the responses of crizotinib therapy. Comparative experiments: (1) Comparison of the results of 4% neutral buffered formalin fixed for different time (24 h, 48 h, 72 h) on the Ventana IHC ALK (D5F3) staining was conducted in two cases of IHC ALK positive FFPE cell blocks; (2) Comparing qRT-PCR results for ALK fusion in samples from FFPE cell blocks and cytospin prepared slides in 10 cases of lung adenocarcinoma cytologic specimens.

RESULTSAmong the specimens examined using the standardized protocol recommended by this study, 229 cases of cytologic specimens met the diagnostic criteria of lung adenocarcinoma. Among them, 207 cases obtained ALK gene test results (by at least one method), with an ALK test ratio of 90.4% (207/229). FFPE cell blocks were successfully prepared in 203 cases, Ventana IHC ALK (D5F3) were successfully performed in all the 203 FFPE cell blocks (100%), and the ALK protein positive detection rate was 10.3% (21/203). ALK fusion was tested in 98 FFPE cytologic samples of EGFR wild types by qRT-PCR, and 96 out of 98 (97.96%) cytologic samples were successfully performed.18 out of 19 IHC ALK-positive cases were verified to be of ALK fusion status by qRT-PCR. The concordance rate was 94.7% (Kappa=0.967, P<0.001) between Ventana IHC ALK (D5F3) and qRT-PCR, and the sensitivity of the Ventana IHC ALK (D5F3) assay compared with qRT-PCR was 100% and the specificity was 98.7%. FISH assay was used to verify the positive cases detected by Ventana IHC ALK (D5F3) staining. Two cases of low tumor cell content FFPE samples obtained indefinite results by FISH test. The six patients with positive ALK protein expression received crizotinib therapy, and 5 paitents got treated effectively. For two ALK IHC positive cases, which were 4% neutral buffered formalin fixed for 72 h, the result of Ventana IHC ALK (D5F3) staining became weakened obviously and uneven. In 10 cases of samples, total RNA was extracted from FFPE cytologic sections and cytospin prepared slides, and the results of qRT-PCR test and ALK gene fusion showed good concordance.

CONCLUSIONSThe standardized protocol recommended in this study expands the detection types and quantity of cytologic specimens for ALK protein expression and gene fusion and increased the detection rate. Ventana IHC ALK (D5F3) is a reliable method for detecting ALK protein expression in FFPE cell blocks. The pathologic quality control procedure prior to Ventana IHC ALK (D5F3) is crucial for the accuracy of testing the ALK gene status. When FFPE cell blocks could not be prepared or prepared unsuccessfully from the cytologic specimens, qRT-PCR may be an alternative option for the detection of ALK gene fusion.

Adenocarcinoma ; drug therapy ; enzymology ; genetics ; pathology ; Alkaline Phosphatase ; genetics ; metabolism ; Gene Fusion ; Genes, erbB-1 ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; drug therapy ; enzymology ; genetics ; pathology ; Protein Kinase Inhibitors ; therapeutic use ; Proteomics ; Pyrazoles ; therapeutic use ; Pyridines ; therapeutic use ; Sensitivity and Specificity

Objective The aim of this study was to establish a standardized protocol for detection of ALK protein expression and gene fusion in cytologic specimens. Methods Lung adenocarcinoma cytologic specimens were collected from seven hospitals in Beijing city. A detection protocol for ALK protein expression and gene fusion was designed according to the results of comparative experiment. Ventana immunohistochemical ( IHC) ALK( D5F3) detecting ALK protein expression was performed in 203 prepared
formalin?fixed paraffin?embedded ( FFPE) cell blocks. ALK gene fusion in 98 EGFR gene wild type cytologic specimens and in 4 bronchoalveolar lavage fluid ( BL ) samples was detected by quantitative reverse transcription polymerase chain reaction (qRT?PCR). ALK gene fusion in the Ventana IHC ALK (D5F3) positive samples was further tested by fluorescence in situ hybridization ( FISH) . Six patients with ALK IHC?positive result were followed up to analyze the responses of crizotinib therapy. Comparative experiments:( 1) Comparison of the results of 4% neutral buffered formalin fixed for different time ( 24 h, 48 h, 72 h) on the Ventana IHC ALK (D5F3) staining was conducted in two cases of IHC ALK positive FFPE cell blocks;(2) Comparing qRT?PCR results for ALK fusion in samples from FFPE cell blocks and cytospin prepared slides in 10 cases of lung adenocarcinoma cytologic specimens. Results Among the specimens examined using the standardized protocol recommended by this study, 229 cases of cytologic specimens met the diagnostic criteria of lung adenocarcinoma. Among them, 207 cases obtained ALK gene test results ( by at least one method), with an ALK test ratio of 90.4% (207/229).FFPE cell blocks were successfully prepared in 203 cases, Ventana IHC ALK ( D5F3) were successfully performed in all the 203 FFPE cell blocks ( 100%) , and the ALK protein positive detection rate was 10.3% (21/203). ALK fusion was tested in 98 FFPE cytologic samples of EGFR wild types by qRT?PCR, and 96 out of 98 ( 97. 96%) cytologic samples were successfully performed.18 out of 19 IHC ALK?positive cases were verified to be of ALK fusion status by qRT?PCR. The concordance rate was 94.7% ( Kappa=0.967, P<0.001) between Ventana IHC ALK( D5F3) and qRT?PCR, and the sensitivity of the Ventana IHC ALK ( D5F3) assay compared with qRT?PCR was 100%and the specificity was 98. 7%. FISH assay was used to verify the positive cases detected by Ventana IHC ALK(D5F3) staining. Two cases of low tumor cell content FFPE samples obtained indefinite results by FISH test. The six patients with positive ALK protein expression received crizotinib therapy, and 5 paitents got treated effectively. For two ALK IHC positive cases, which were 4% neutral buffered formalin fixed for 72 h, the result of Ventana IHC ALK(D5F3) staining became weakened obviously and uneven. In 10 cases of samples, total RNA was extracted from FFPE cytologic sections and cytospin prepared slides, and the results of qRT?PCR test and ALK gene fusion showed good concordance. Conclusions The standardized protocol recommended in this study expands the detection types and quantity of cytologic specimens for ALK protein expression and gene fusion and increased the detection rate. Ventana IHC ALK( D5F3) is a reliable method for detecting ALK protein expression in FFPE cell blocks. The pathologic quality control procedure prior to Ventana IHC ALK( D5F3) is crucial for the accuracy of testing the ALK gene status. When FFPE cell blocks could not be prepared or prepared unsuccessfully from the cytologic specimens, qRT?PCR may be an alternative option for the detection of ALK gene fusion.

Objective The aim of this study was to establish a standardized protocol for detection of ALK protein expression and gene fusion in cytologic specimens. Methods Lung adenocarcinoma cytologic specimens were collected from seven hospitals in Beijing city. A detection protocol for ALK protein expression and gene fusion was designed according to the results of comparative experiment. Ventana immunohistochemical ( IHC) ALK( D5F3) detecting ALK protein expression was performed in 203 prepared
formalin?fixed paraffin?embedded ( FFPE) cell blocks. ALK gene fusion in 98 EGFR gene wild type cytologic specimens and in 4 bronchoalveolar lavage fluid ( BL ) samples was detected by quantitative reverse transcription polymerase chain reaction (qRT?PCR). ALK gene fusion in the Ventana IHC ALK (D5F3) positive samples was further tested by fluorescence in situ hybridization ( FISH) . Six patients with ALK IHC?positive result were followed up to analyze the responses of crizotinib therapy. Comparative experiments:( 1) Comparison of the results of 4% neutral buffered formalin fixed for different time ( 24 h, 48 h, 72 h) on the Ventana IHC ALK (D5F3) staining was conducted in two cases of IHC ALK positive FFPE cell blocks;(2) Comparing qRT?PCR results for ALK fusion in samples from FFPE cell blocks and cytospin prepared slides in 10 cases of lung adenocarcinoma cytologic specimens. Results Among the specimens examined using the standardized protocol recommended by this study, 229 cases of cytologic specimens met the diagnostic criteria of lung adenocarcinoma. Among them, 207 cases obtained ALK gene test results ( by at least one method), with an ALK test ratio of 90.4% (207/229).FFPE cell blocks were successfully prepared in 203 cases, Ventana IHC ALK ( D5F3) were successfully performed in all the 203 FFPE cell blocks ( 100%) , and the ALK protein positive detection rate was 10.3% (21/203). ALK fusion was tested in 98 FFPE cytologic samples of EGFR wild types by qRT?PCR, and 96 out of 98 ( 97. 96%) cytologic samples were successfully performed.18 out of 19 IHC ALK?positive cases were verified to be of ALK fusion status by qRT?PCR. The concordance rate was 94.7% ( Kappa=0.967, P<0.001) between Ventana IHC ALK( D5F3) and qRT?PCR, and the sensitivity of the Ventana IHC ALK ( D5F3) assay compared with qRT?PCR was 100%and the specificity was 98. 7%. FISH assay was used to verify the positive cases detected by Ventana IHC ALK(D5F3) staining. Two cases of low tumor cell content FFPE samples obtained indefinite results by FISH test. The six patients with positive ALK protein expression received crizotinib therapy, and 5 paitents got treated effectively. For two ALK IHC positive cases, which were 4% neutral buffered formalin fixed for 72 h, the result of Ventana IHC ALK(D5F3) staining became weakened obviously and uneven. In 10 cases of samples, total RNA was extracted from FFPE cytologic sections and cytospin prepared slides, and the results of qRT?PCR test and ALK gene fusion showed good concordance. Conclusions The standardized protocol recommended in this study expands the detection types and quantity of cytologic specimens for ALK protein expression and gene fusion and increased the detection rate. Ventana IHC ALK( D5F3) is a reliable method for detecting ALK protein expression in FFPE cell blocks. The pathologic quality control procedure prior to Ventana IHC ALK( D5F3) is crucial for the accuracy of testing the ALK gene status. When FFPE cell blocks could not be prepared or prepared unsuccessfully from the cytologic specimens, qRT?PCR may be an alternative option for the detection of ALK gene fusion.

OBJECTIVEThe aim of this study was to establish a standard protocol for detection of EGFR mutations in cytologic specimens.

METHODS287 cytologic samples were collected from the patients who were suspected of having lung cancer at six hospitals in Beijing. A detection protocol for EGFR mutations was designed. Two comparative experiments were carried out for the coincidence in EGFR mutation rates between direct sequencing (Seq) and amplification refractory mutation system (ARMS) methods, and between 40 matched cytologic samples with formaldehyde-fixed paraffin embedded (FFPE) cytologic blocks and cytospin slides.

RESULTSTumor cells were found in 236 out of 287 cases (82.2%, 236/287) . Among them, there were 31 cases (13.1%, 31/236) of low tumor cell content samples and 205 cases (86.9%, 205/236) of high tumor cell content samples. 180 cases in the high tumor cell content samples (87.8%, 180/205) were diagnosed to be consistent with NSCLC. 25 out of 194 cases were ruled out or indefinite to be diagnosed as NSCLC by immunohistochemistry. By direct sequencing, the mutation rate of EGFR was 27.8% (50/180) in NSCLC samples and 28.2% (50/177) in adenocarcinoma samples (high tumor content samples) . By ARMS, the mutation rate of EGFR was 45.6% (82/180) in NSCLC samples and 46.3% (82/177) in adenocarcinoma samples (high tumor content samples). The EGFR mutation rate in low tumor content samples was 38.7% (12/31) , there was no significant difference in EGFR mutation rates between the groups of low tumor cell content samples and high tumor cell content samples (P = 0.12). The concordance rate of EGFR mutation rates was 100% between scraping tumor cells from slides samples and from FFEP blocks in the 40 matched samples. Forty-eight out of 180 definitive NSCLC patients received Gefitinib therapy. The FPS was 12 months in the gefitinib-treated ARMS⁺ group and 2 months in the ARMS⁻ group (P < 0.001), and the OS was 19 months in the gefitinib-treated ARMS⁺ group and 7 months in the ARMS⁻ group (P = 0.003), but no significant differences were found in the efficacy (PFS and OS) of Gefitinib between Seq⁺ and Seq⁻ groups (P = 0.227, P = 0.510, respectively), and Seq⁺/ARMS⁺ and Seq⁻/ARMS⁺ groups (P = 0.354, P = 0.334, respectively).

CONCLUSIONSThe detection protocol for EGFR mutations in cytological specimens introduced in this study is tested to be reliable and feasible. Pathological evaluation and immunohistochemistry are important in the detection procedure of EGFR mutations in cytologic specimens. High sensitivity methods should be selected for detection of EGFR mutations in cytologic samples.

Adenocarcinoma ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; Humans ; Lung Neoplasms ; diagnosis ; epidemiology ; metabolism ; Mutation ; Mutation Rate ; Polymerase Chain Reaction ; Receptor, Epidermal Growth Factor ; genetics ; metabolism

Objective To evaluate the medical indexes for student pilots from different areas , to discover the major different indexes between different areas ,and to establish the space distribution model of military pilots .Methods A cross-section survey was conducted among student pilots , and 290 student pilots sampled as respondents were interviewed with questionnaires and subjected to a physical examination , involving distant vision , heart function , and pulmonary function , before a database was established , cleaned and analyzed by EpiData 3.02, SAS 9.13 with double checking .Results There was no difference between the medical indexes of student pilots from 7 areas, but the psychological selection performance record and the entrance examination record were different .Student pilots from area E had the highest psychological selection performance record while those from area D had the highest entrance examination record .Conclusion Student pilots have area difference ,so we should pay close attention to their birth place during recruitment of student pilots .