[Clinicopathological and genetic characteristics of congenital cystic adenomatoid malformation of lung and its associated lung cancer in adults].

Objective: To investigate the clinicopathological features and genetic characteristics of congenital cystic adenomatoid malformation (CCAM) of lung and CCAM associated lung cancer in adults. Methods: A total of 13 cases of CCAM of lung in adults, diagnosed from June 2015 to May 2023, were collected from the Department of Pathology, Shanghai Pulmonary Hospital, Tongji University School of Medicine, China. Their histopathological features were correlated with probable development into lung cancer. Next-generation sequencing was performed on the benign and malignant areas of all cases. Results: The pathological classification of all cases were of CCAM of lung type 1. There were 4 male and 9 female cases, age ranged from 18 to 65 years, with a mean age of 41 years. Six cases were accompanied by lung cancer, all of them were mucinous adenocarcinoma. Next-generation sequencing showed no gene mutation in 2 of the 13 cases; KRAS mutations in exon 2 were detected in 7 cases, in which there were 6 cases complicated with lung mucinous adenocarcinoma and no matter in the malignant or benign regions, the same case exhibited the same mutation sites in KRAS gene. Conclusions: CCAM of the lung is a congenital disease, and in adults, type 1 is most commonly found in the pathological classification, and it is often accompanied by cancer. Gene mutations are frequently detected in CCAM of the lung, KRAS being the most recurrent mutation which may play an important role in the carcinogenesis.

[Application of the WHO Classification of Thoracic Tumors (2021) grading system in invasive pulmonary adenocarcinoma and its correlation with the targeted genes' variations].

Objective: To investigate the applicability of the 2021 WHO classification of thoracic tumors' new grading system for invasive pulmonary adenocarcinoma (IPA) with different clinical stages and its correlation with the characteristics of targeted genes' variation. Methods: A total of 2 467 patients with surgically resected primary IPA in Shanghai Pulmonary Hospital, Shanghai, China from September to December 2020 were retrospectively analyzed. Eligible cases were graded using the new grading system of IPA of the 2021 WHO classification of thoracic tumors. The clinicopathological data and targeted-gene abnormality were collected. The utility of new grading system of IPA in different clinical stages was investigated. The correlation of clinicopathological features and targeted-gene abnormality in different grades of IPA were compared. Results: All 2 311 cases of IPA were included. There were 2 046 cases of stage Ⅰ IPA (88.5%), 169 cases of stage Ⅱ (7.3%), and 96 cases of stage Ⅲ (4.2%). According to the new classification system of IPA, 186 cases (9.1%), 1 413 cases (69.1%) and 447 cases (21.8%) of stage-Ⅰ adenocarcinoma were classified as Grade 1, Grade 2 and Grade 3, respectively. However, there were no Grade 1 adenocarcinomas in stages Ⅱ and Ⅲ cases. Among stage-Ⅱ and Ⅲ IPA cases, there were 38 Grade 2 cases (22.5%) and 131 Grade 3 cases (77.5%), and 3 Grade 2 cases (3.1%) and 93 Grade 3 cases (96.9%), respectively. In stage-Ⅰ cases, no tumor cells spreading through airspace (STAS), vascular invasion or pleural invasion was found in Grade 1 of IPA, while the positive rates of STAS in Grade 2 and 3 IPA cases were 11.3% (159/1 413) and 73.2% (327/447), respectively. There was a significant difference among the three grades (P<0.01). Similarly, the rates of vascular and pleural invasion in Grade 3 IPA cases were 21.3% (95/447) and 75.8% (339/447), respectively, which were significantly higher than those of 1.3% (19/1 413) and 3.0% (42/1 413) in Grade 2 (P<0.01). EGFR mutational rates in Grades 1, 2 and 3 IPA were 65.7% (94/143), 76.4% (984/1 288) and 51.3% (216/421), respectively. The differences among the three grades were statistically significant (P<0.01). No fusion genes were detected in Grade 1 IPA, while the positive rates of ROS1 and ALK fusion genes in Grade 3 were 2.4% (10/421) and 8.3% (35/421), respectively, which were significantly higher than that of 0.5% (7/1 288) and 1.6% (20/1 288) in Grade 2 (P<0.01). In stage-Ⅱ cases, only EGFR mutation rate in Grade 2 adenocarcinoma (31/37, 83.8%) was higher than that in Grade 3 adenocarcinoma (71/123, 57.7%; P<0.01). However, the correlation between the new grade system of IPA and the distribution characteristics of targeted-gene variation cannot be evaluated in stage Ⅲ cases. Conclusions: The new grading system for IPA is mainly applicable to clinical stage-Ⅰ patients. Tumor grades of IPA are strongly correlated with the high-risk factors of prognosis and the distribution features of therapeutic targets. It is of great significance and clinical value to manage postoperative patients with early-stage IPA.

[Clinicopathological and genetic characteristics of bronchial sialadenoma papilliferum: report of four cases].

Objective: To investigate the clinicopathological, immunophenotypic, and molecular genetic features of bronchial sialadenoma papilliferum (BSP). Methods: Four cases of BSP collected at the Shanghai Pulmonary Hospital from May 2018 to June 2021 were retrieved and analyzed. These cases were evaluated for their clinical, histological, immunohistochemical (IHC) and genomic features. The patients were followed up and relevant literature was reviewed. Results: All four patients were male, aged from 55 to 75 years (mean 62 years), with tumor diameter of 6 to 21 mm (mean 13.5 mm), and lesions were located in the left lower lobe (n=2), right lower lobe (n=1), and trachea (n=1). They were characterized by a combination of surface exophytic endobronchial papillary proliferation and an endophytic two-cell layered ductal structure. IHC staining showed that CK7 and EMA were strongly positive in ductal epithelium; p63, p40, CK5/6 were positive in ductal and papillary basal cells; SOX10 was positive in ductal epithelium and basal cells; S-100 was positive in basal cells and ductal epithelium in two cases. Next generation sequencing showed that two cases harbored BRAF V600E mutation. Conclusions: BSP is an extremely rare primary lung tumor arising from the salivary gland under bronchial mucosa. The primary treatment choice of this tumor is complete surgical resection. The diagnosis and differential diagnosis of this tumor depend on classic histomorphologic and IHC features, and BRAF V600E gene mutation can be detected.

[Pathological assessment of non-small cell lung cancer resection specimens after neoadjuvant therapy].

Objective: To pathologically evaluate the surgically resected specimens of three different therapies (neoadjuvant chemotherapy, neoadjuvant targeted therapy and neoadjuvant immunotherapy combined with chemotherapy) for non-small cell lung cancer. Methods: One-hundred and thirteen cases of post neoadjuvant therapy non-small cell lung cancer specimens were collected at Tongji University Affiliated Shanghai Pulmonary Hospital from January 2000 to March 2020. There were ninty patients receiving neoadjuvant chemotherapy (chemotherapy group;26 cases of adenocarcinoma and 64 cases of squamous cell carcinoma), 13 patients receiving neoadjuvant targeted therapy (targeted group;13 cases of adenocarcinoma) and 10 patients receiving neoadjuvant immunotherapy combined with chemotherapy (immune combined chemotherapy group;4 cases of adenocarcinoma and 6 cases of squamous cell carcinoma). They were evaluated for histologic tumor regression responses (necrosis, inflammatory cell infiltration, cholesterol crystal deposition, foam cell infiltration, reactive granuloma and interstitial collagenous formation) and pathological responses [main pathological response (MPR) and complete pathological response (PCR)]. Results: Chemotherapy group, targeted group and immune combined chemotherapy group all showed degenerative changes in residual tumor cells, increased atypia, various degrees of necrosis, foam cell aggregation, cholesterol cleft, inflammatory cell infiltration, and reactive granuloma in the tumor bed. Histologic characteristics of tumor regression reaction were not different between these three groups (P>0.05); the highest percentage of necrosis in the targeted group and immune combined chemotherapy group was only 10% and 20%, respectively, while that in the chemotherapy group was as high as 80%. One case of adenocarcinoma in immune combined chemotherapy group had tumor regression bed. The MPR rates of adenocarcinoma in chemotherapy group and squamous cell carcinoma in chemotherapy group were 35% (9/26) and 64% (41/64), respectively; the MPR ratio of targeted group was 2/13; the MPR ratio of adenocarcinomain immune combined chemotherapy group and squamous cell carcinoma in immune combined chemotherapy group were 2/4 and 2/6, respectively. The PCR rates of adenocarcinoma in chemotherapy group and squamous cell carcinoma in chemotherapy group were 11% (3/26) and 3% (2/64), respectively; the PCR ratio of targeted group was 0/13; the PCR ratio of adenocarcinomain immune combined chemotherapy group and squamous cell carcinomain immune combined chemotherapy group were 0/4 and 1/6, respectively. Conclusions: Different neoadjuvant therapy may cause various histopathological changes in non-small cell lung cancer: more necrosis is noted in the chemotherapy group and regression bed frequently appears in the immune combined chemotherapy group. In the immune combined chemotherapy group, there are significant lymphoplasmacytic infiltration and lymphoid follicle formation in the lung parenchyma beside the tumor bed.

[Clinicopathologic features and genetic profile of the redefined large cell lung carcinoma].

Objective: To investigate the clinicopathologic features and genetic profile of large cell lung carcinoma (LCC) redefined by new classification. Methods: Basing on 2015 WHO classification criteria in redefining large cell lung carcinoma, the expression of specific markers (TTF1, Napsin A, p40, CK5/6, CK, vimentin and ZEB1) was detected by immunohistochemistry and D-PAS staining in 303 surgically-removed lung specimens previously diagnosed as large cell lung carcinoma. The clinicopathologic and genetic characteristics (including EGFR, KRAS, BRAF, ALK and ROS1 gene mutation) were analyzed. Results: Based on the new definition of LCC, 116 cases (116/303, 38.3%) of LCC formerly diagnosed were reclassified as solid adenocarcinoma, 49 cases (49/303, 16.2%) as squamous cell carcinoma, 6 cases (6/303, 2.0%) as adenosquamous carcinoma, 22 cases (22/303, 7.3%) as spindle cell carcinoma and only 110 cases (110/303, 36.3%) as large cell carcinoma. Redefined LCCs were characterized as middle-age (range 40-80), male (102/110, 92.7%) and smoking patients (64/110, 58.2%) with intermediate-advanced stage. Among 110 cases, 9 cases with EGFR mutation and 10 cases with KRAS mutation and 1 case with ALK fusion were found. No BRAF and ROS1 alterations were identified. Conclusions: According to the new classification, LCCs formerly diagnosed are mostly reclassified as adenocarcinoma and non-keratinizing squamous cell carcinoma. The newly defined LCC may significantly benefit from clinical therapy.

[Research advances in the v-raf murine sarcoma viral oncogene homolog gene mutation in ameloblastoma].

V-raf murine sarcoma viral oncogene homolog B1 (BRAF) is a pro-oncogene, which is one member of the RAF family. Mutated BRAF is found in approximately 8% of human tumors. BRAF gene mutations lead to continuous activation of the mitogen-activatd protein kinase (MAPK) pathway, which resulting in abnormal cell proliferation and tumorigenesis. In recent years, recurrent MAPK signaling mutations were identified in ameloblastoma, among which BRAF-V600E is the most prominent type. This provides new strategies for the targeted treatment of ameloblastoma. This paper reviewed the latest advances in BRAF gene mutation associated with ameloblastoma and its potential clinical significance.

Prospective study of serum selenium levels and incident esophageal and gastric cancers.

BACKGROUND: From March 1986 through May 1991, we conducted a randomized nutritional intervention trial, the General Population Trial, in Linxian, China, a region with epidemic rates of squamous esophageal and adenomatous gastric cardia cancers. We found that participants who received selenium, beta-carotene, and vitamin E had significantly lower cancer mortality rates than those who did not. In the current study, we examined the relationship between selenium levels measured in pretrial (1985) sera from participants and the subsequent risk of developing squamous esophageal, gastric cardia, and gastric non-cardia cancers during the trial. METHODS: This study was designed and analyzed in accord with a stratified case-cohort sampling scheme, with the six strata defined by sex and three age categories. We measured serum selenium levels in 590 case subjects with esophageal cancer, 402 with gastric cardia cancers, and 87 with gastric non-cardia cancers as well as in 1062 control subjects. Relative risks (RRs), absolute risks, and population attributable risk for cancers were estimated on the basis of the Cox proportional hazards models. All statistical tests are two-sided. RESULTS: We found highly significant inverse associations of serum selenium levels with the incidence of esophageal (P: for trend <10(-4)) and gastric cardia (P: for trend <10(-6)) cancers. The RR and 95% confidence interval (CI) for comparison of highest to lowest quartile of serum selenium was 0.56 (95% CI = 0.44-0.71) for esophageal cancer and 0.47 (95% CI = 0.33-0.65) for gastric cardia cancer. The population proportion of these cancers that is attributable to low selenium levels was 26.4% (95% CI = 14.45-38.36). We found no evidence for a gradient of serum selenium associated with incidence of gastric non-cardia cancer (P: for trend =.96), with an RR of 1.07 (95% CI = 0.55-2.08) for the highest to lowest quartile of serum selenium. CONCLUSIONS: Our study supports findings from previous prospective studies and randomized trials that variations in selenium levels affect the incidence of certain cancers. In the United States, where intervention trials of selenium are in the planning stages, consideration should be given to including populations at high risk for squamous esophageal and gastric cardia cancers.

Significance of lipopolysaccharide detection in children with pulmonary infections.

OBJECTIVE: The present study aimed to observe the alterations of lipopolysaccharide (LPS) and some other laboratory indexes in children suffering from pulmonary infections, and to investigate the condition of Gram-negative bacterial infection. PATIENTS AND METHODS: All the patients received routine blood test, C reactive protein (CRP), erythrocyte sedimentation rate (ESR), Mycoplasma pneumoniae antibody IgM (MP-IgM), LPS, blood culture and chest X-ray examination. The clinical data was collected followed by induction arrangement and statistical analysis. RESULTS: In terms of the rate of abnormity in peripheral white blood cell count and positive rate of blood CRP, no significant difference was found between children with pulmonary infections and the healthy individual in the control group (p > 0.05). The positive rates of blood MP-IgM were 33.33% and 32.26% in children with different progressive stages of pulmonary infections, which were significantly lower than those in the control group (62.96%) (p < 0.05). The positive rates of blood LPS in the observation group were higher than those in the control group, especially for those children at progressive stages within one week; and the difference between them was significant (p < 0.05). With regard to blood bacterial culture, the positive rates were 9.52% and 29.03% for children in progressive stages within one week and over one week in the observation group, respectively; the latter was significantly higher than that in the control group (p < 0.05). The result of the correlation analysis suggested a weak correlation between the positive rate of increased blood LPS in the observation group and that in blood bacterial culture (χ2 = 6.61, p < 0.05; Pearson's contingency coefficient C = 0.34). However, there was no significant correlation between the positive rate of increased blood LPS and peripheral blood white cell count, CRP, or MP-IgM (p > 0.05). CONCLUSIONS: Endotoxemia is often accompanied by pulmonary infections, and gram-negative bacterium is a common pathogenic bacterium in children with different progressive stages of pneumonia.

Detection of congenital cytomegalovirus infection by using chorionic villi of the early pregnancy and polymerase chain reaction.

OBJECTIVE: To detect congenital cytomegalovirus (CMV) infection of chorionic villi in early pregnancy. METHODS: Extraction of DNA of chorionic villi and amplification of the gene of major immediate-early (MIE) antigen of CMV using a polymerase chain reaction (PCR). RESULTS: Sixty-eight specimens of chorionic villi and 16 specimens were positive for CMV infection by PCR. The incidence of congenital CMV infection in the first trimester of pregnancy was 23.5%. CONCLUSIONS: The risk of transmission of CMV from mother to fetus in early pregnancy is very high and potential CMV carriers may transmit CMV to their fetus in early pregnancy.

Detection of Chlamydia trachomatis intrauterine infection using polymerase chain reaction on chorionic villi.

OBJECTIVE: To detect Chlamydia trachomatis intrauterine infection in the early pregnancy by using chorionic villi. METHOD: The C. trachomatis infection in pregnant women was investigated by cervical specimens and Clearview kits. DNA of chorionic villi was extracted and the gene of a major outer membrane protein of C. trachomatis was amplified by polymerase chain reaction (PCR). RESULTS: 120 cervical specimens of the pregnant women were analyzed and 10 cervical specimens were positive for C. trachomatis infection. In this study, the prevalence of C. trachomatis infection was approx. 8.3%. Fifty-nine specimens of chorionic villi and three positive specimens of C. trachomatis infection were analyzed by PCR. The incidence of C. trachomatis intrauterine infection in the early pregnancy was 5.1%. CONCLUSION: The vertical transmission of C. trachomatis infection in the early pregnancy may be a pathway of intrauterine infection. Chorionic villus sampling in early pregnancy and the PCR method could be developed as a technique for prenatal diagnosis of C. trachomatis intrauterine infection.

Detection of a human parvovirus intrauterine infection with the polymerase chain reaction.

OBJECTIVE: To detect human parvovirus B19 intrauterine infection in pregnancy with the polymerase chain reaction (PCR). STUDY DESIGN: DNA of chorionic villi and amniotic fluid was extracted and the gene of human parvovirus B19 amplified with PCR. RESULTS: The study analyzed 61 specimens of chorionic villi and 26 specimens of amniotic fluid and found two positive specimens of chorionic villi and 1 positive specimen of amniotic fluid. CONCLUSION: The vertical transmission of human parvovirus B19 infection in early pregnancy may be a pathway of intrauterine infection. Chorionic villus sampling in early pregnancy and PCR could be developed as a method of prenatal diagnosis of human parvovirus B19 intrauterine infection.

Anti-FD4 idiotypic antibody mimicking human gastric cancer-associated antigen.

Rabbits were immunized with the Fab fragment of a murine monoclonal antibody (McAb) PD4 against human gastric cancer to produce anti-PD4-idiotypic antibody (alpha PD4-Ab2). The alpha PD4-Ab2 could not only competitively inhibit binding of McAb PD4 to gastric cancer cell MGC803, but also induce delayed-type hypersensitivity (DTH) to MGC803 in mice. Spleen cells of mice immunized with alpha PD4-Ab2 were fused with myeloma cell SP2/0 to form hybridoma secreting Ab3 which could be bound to target cell MGC803. McAb C7-Ab3, one of the Ab3, could selectively react with a 40 kD tumor-associated antigen located on MGC803 cell membrane, as well as McAb PD4. The results indicate that alpha PD4-Ab2 possesses determinants (internal image antigen) similar to those on MGC803, and could mimic human gastric cancer-associated antigen.

[Preparation and identification of monoclonal antibody against the gastric cancer cell line MGC 803].

Spleen cells of Balb/c mice, immunized with gastric cancer cell MGC 803, were fused with murine myeloma cell NS-1. After selective culture, screening and subcloning, a hybridoma PC1 which produced monoclonal antibody (McAb) against MGC 803 cells was obtained. McAb PC1 bound strongly with 3/4 gastric cancer and 1/2 hepatoma cell lines, weakly with another gastric cancer and 2/2 lung cancer cell lines, but did not bind with the autologous and allogenic lymphocytes, ABO red blood cells, human fetal lung fibroblasts and normal bone marrow cells. The binding capacity of McAb PC1 to MGC 803 decreased significantly due to the absorption by MGC 803 cells, but was not affected by lymphocytes and CEA. The corresponding antigen of McAb PC1 was expressed on the surface of MGC 803 cells. It may be a gastric cancer-associated antigen.

[Gastric cancer-associated antigen P42].

The corresponding antigen to monoclonal antibody PCI against gastric cancer cells was demonstrated as a protein sensitive to heating and easily degraded by proteinase. By western blotting, it was shown that the molecular weight of the antigen was 42 KD and the antigen was named P42. P42 could be shed into serum-free culture medium of the target cells and its activity be assayed in sera of the gastric cancer patients. Using inhibition test, the levels of P42-like substance in sera were measured. The results indicated that this level in the gastric cancer patients (26.53 +/- 23.11) was markedly higher than that in patients with benign gastric diseases (4.66 +/- 2.67) and controls (2.57 +/- 2.67). It is suggested that P42, a novel gastric cancer-associated antigen, be useful in the mass screening and supplementary in diagnosing gastric cancer.

[Expression of the surface antigen in human gastric cancer cells and the relation to cell cycles--correlated analysis with flow cytometry].

Expression of tumor-associated antigen in different gastric cancer cell lines and different phases of cell cycle was studied cytochemically. The antigen was recognized by the monoclonal antibody (McAb) PC1 against gastric cancer cells. By using the McAb PC1 as first antibody, the indirect immunofluorescence stain and the peroxidase-anti-peroxidase (PAP) stain were done on the gastric cancer cell lines (MGC 803, SGC 7901 and BGC 823). It was shown that PC1 antigen was mainly expressed on the membrane of these cells and only a certain percentage of the cells gave the positive reaction with different intensities. It was obvious that the expression of PC1 antigen was heterogeneous in nature. The heterogeneity of the PC1 antigen expression in gastric cancer cells might be due to either various subpopulations in the cell lines or different phases of cell cycle. In order to go further into the question, we studied quantitatively the expression of PC1 antigen in gastric cancer cell lines (MGC 803, STC 7901 and BGC 823) and the relationship between the antigen expression and cell cycle by double fluorescence stain and two-dimensional flow cytometry. It was found that expression levels of PC1 antigen in these cell lines were in the following order: MGC 803 greater than SGC 7901 greater than BGC 823. The PC1 antigen predominantly expressed on G1 phase for MGC 803 and G1, G2-M phase for SGC 7901 respectively. And uniform low level of PC1 antigen expression was found for BGC 823 throughout the cell cycle. Therefore, the PC1 antigen expression is dependent on cell cycle in MGC 803 and SGC 7901 cell lines.

[Assessment of the mutagenic effect of maternal factors on human chorionic villi by micronucleus test].

Rapid determination of DNA damage by micronucleus test is well accepted. Animal bone marrow cells or human peripheral lymphocytes used in most studies could not directly reflect the influence of the mutagenic effect on the offsprings by environmental factors. Human chorionic villi micronucleus test to detect directly the mutagenic effect of environmental factors has not been reported in the literature. Direct determination of human chorionic villi micronuclei (CVMN) was established in our laboratory, to study the mutagenic effect of mother's age, gravidity, gestation age, abortion history, contraception (condom, diaphragm, rhythm, oral contraceptives, spermicide or IUD), smoking and drinking on the offsprings. Cross investigation and micronucleus test were used in 507 couples undergoing artificial abortion. Micronuclei were scored according to Countryman's standard 2,000 interphases were observed in each subject for CVMN frequency (%). Arcsine transformation (arcsine [Sqr (P)]) was used in transforming CVMN frequency and the analysis of variance were used for statistics. No correlation between CVMN frequency and mother's age, gravidity, gestation age, abortion history, and contraception was found. Neither smoking nor drinking habit was found among the women of this study. The CVMN frequency of husband smoking was 0.7645 +/- 0.0561%, of husband non-smoking-drinking was 0.5522 +/- 0.0616%, of husband drinking was 0.5667 +/- 0.2004%, of husband smoking and drinking was 0.7944 +/- 0.0754%. There was a statistical difference in CVMN frequency between husband smoking and non-smoking (F = 2.78 DF = 408 P less than 0.05). No significant difference was found between husband drinking and non-drinking.