Objective: To investigate the prevalence of BRAF V600E mutation in thyroid nodules and to analyze the relationship between BRAF V600E mutation and various clinicopathological features. Methods: BRAF V600E mutant gene test was done in 463 cases of thyroid nodules collected from April 2015 to July 2018 in Beijing Hospital. Pathologic sections of 444 cases of papillary thyroid carcinoma were reviewed and clinical information was collected.Statistical analysis of the relationship between BRAF V600E gene mutation and various clinicopathological features was performed with SPSS 21.0 statistical software. Results: There were 109 males and 354 females in the cohort, with a male to female ratio of 1.0∶3.2. The patient ranged in age from 16 to 82 years, with an average age of 46.1 years. The BRAF V600E mutation rates in papillary thyroid carcinoma, benign thyroid nodules and other thyroid carcinoma were 86.5%(384/444),0/15 and 1/4,respectively.There was significant correlation between BRAF V600E mutation and histological diagnosis of papillary thyroid carcinoma (P<0.05). There was no correlation with age, gender, multifocality, bilaterality, coexisting lymphocytic thyroiditis, nodular goiter, maximum diameter, capsule invasion, extrathyroidal extension and clinical stage (P>0.05). Conclusions BRAF V600E gene mutation is closely related to the occurrence of papillary thyroid carcinoma. BRAF V600E has significant value in the diagnosis of papillary thyroid carcinoma. While BRAF V600E mutation is related to the histological diagnosis, it shows no correlation with other clinicopathologic features. BRAF V600E mutation is not an independent prognostic factor in papillary thyroid carcinoma patients.

Objective To investigate the expression of Girdin and its significance in Alzheimer′s disease ( AD).Methods Fifty-nine autopsy cases from Department of Pathology, Beijing Hospital from January 1988 to December 2013, including 35 AD cases and 24 non-AD cases as control.Girdin and amyloid β-protein ( Aβ) expression was evaluated immunohistochemically by EnVision method.The correlation between Girdin and Aβwas analyzed.Results Girdin expression was localized in the nucleus and/or cytoplasm.The expression rates of Girdin were 20.0% ( 7/35 ) in the AD group and 83.3%(20/24) in the non-AD group, respectively.The difference was significant ( Yates′s correction for continuityχ2 =20.527, P<0.05).Girdin expression and Aβdeposition also correlated significantly ( P<0.05).Conclusions Girdin shows reduced expression in AD, and is correlated positively with Aβdeposition.This suggests that Girdin may play an important role in the occurrence and development of AD.

Objective To investigate the prevalence of BRAF V600E mutation in thyroid nodules and to analyze the relationship between BRAF V600E mutation and various clinicopathological features. Methods BRAF V600E mutant gene test was done in 463 cases of thyroid nodules collected from April 2015 to July 2018 in Beijing Hospital. Pathologic sections of 444 cases of papillary thyroid carcinoma were reviewed and clinical information was collected.Statistical analysis of the relationship between BRAF V600E gene mutation and various clinicopathological features was performed with SPSS 21.0 statistical software. Results There were 109 males and 354 females in the cohort, with a male to female ratio of 1.0∶3.2. The patient ranged in age from 16 to 82 years, with an average age of 46.1 years. The BRAF V600E mutation rates in papillary thyroid carcinoma, benign thyroid nodules and other thyroid carcinoma were 86.5%(384/444),0/15 and 1/4,respectively.There was significant correlation between BRAF V600E mutation and histological diagnosis of papillary thyroid carcinoma(P<0.05). There was no correlation with age, gender, multifocality, bilaterality, coexisting lymphocytic thyroiditis, nodular goiter, maximum diameter, capsule invasion, extrathyroidal extension and clinical stage (P>0.05). Conclusions BRAF V600E gene mutation is closely related to the occurrence of papillary thyroid carcinoma. BRAF V600E has significant value in the diagnosis of papillary thyroid carcinoma. While BRAF V600E mutation is related to the histological diagnosis, it shows no correlation with other clinicopathologic features. BRAF V600E mutation is not an independent prognostic factor in papillary thyroid carcinoma patients.

Objective:To study the application of cell transfer technology to solve the problem of the limited number of fine needle aspiration cytology (FNAC) smears for various immunocytochemistry (ICC) staining and other auxiliary tests, and to enhance accurate cytological diagnosis.Methods:Thirty-four cases of FNAC smears from January 2020 to April 2020 in the Department of Pathology of Beijing Hospital were collected for investigation of the cell transfer technique. The materials in the most cell smear were divided and transferred to several glass slides. After de-staining, the recipient slides were stained with EnVision ICC. The technique was validated by comparing the consistency of the ICC of transferred cell smears and the corresponding immunohistochemical (IHC) staining on biopsies.Results:There were a total of 180 cell transfer slides from 34 cases, of which 174 had the same cell morphology, size and structure as the original smears, with the success rate of cell transfer of 96.7% (174/180). Totally 174 ICC stains were performed on the successfully transferred cell smears, of which 153 smears had available corresponding IHC staining of histologic specimens. Of these, 148 showed concordance between ICC staining and the IHC staining. Cells were successfully transferred in 96.7 % (148/153) of the cell sheets, keeping the same morphology and structure as compared to their original smears. The diagnosis of all 34 FNAC cases was the same to that of their corresponding pathology on biopsies with 100 % concordance.Conclusions:The cell transfer technique is a simple and effective way to make full use of diagnostic cells on a cell smear, and is valuable for accurate cytological diagnosis.

OBJECTIVE: To develop a cell-based system for the diagnosis of vitamin K-dependent coagulation factor deficiency 1 (VKCFD1). METHODS: In HEK293 cells stably expressing the reporter gene FIX-Gla-PC, the gamma-glutamyl carboxylase (GGCX) gene was knocked out by using CRISPR/Cas9 technology. Enzyme-linked immunosorbent assay (ELISA), DNA sequencing and Western blotting were used to identify the GGCX gene knockout cells. A quickchange point variant method was used to construct the GGCX variant. ELISA was used to assess the influence of GGCX variant on the activity of reporter gene. RESULTS: Two monoclonal cell lines with no reporter activity by ELISA was identified. Edition and knockout of the GGCX gene was confirmed by DNA sequencing and Western blotting. The activity of the reporter gene was recovered by transfection of the wild-type GGCX gene. Thereby two monoclonal cells with GGCX knockout were obtained. By comparing the wild-type and pathogenic GGCX variants, the reporter activity was decreased in the pathogenic variants significantly. CONCLUSION A cell-based system for the detection of GGCX activity was successfully developed, which can be used for the diagnosis of VKCFD1 caused by GGCX variants.

Objective:To investigate the diagnostic value of the combination of 18F-prostate specific membrane antigen (PSMA) PET/CT and multiparametric magnetic resonance imaging (mpMRI) in identifying the grade group of prostate cancer, using parameters derived from the two imaging modalities. Method:Prostate cancer patients diagnosed by histopathology and received 18F-PSMA PET/CT and mpMRI during September 2018 to May 2021 in our hospital were retrospectively studied. The median age was 68(64-75), with the median PSA level of 14.74(7.75-24.19)ng/mL. All patients received mpMRI before biopsy. On biopsy, 6(12.2%) patients had International Society of Urological Pathology grade group(ISUP GG) 1 diseases, 16(32.7%) had ISUP GG 2 diseases, 12(24.5%) had ISUP GG 3 diseases, and 15(10.9%) had ISUP GG 4 or 5 diseases. Patients were then divided into high-grade group (ISUP 4-5) and low-grade group(ISUP 1-3). The median age of patients in high-grade group and low-grade group were 65(62-76) and 71(65-74), respectively. The PSA level in high-grade group and low-grade group were 15.11(6.63-42.86) ng/ml and 12.31(7.94-18.25) ng/ml, respectively. No significant differences were found in age and PSA level between the two groups ( P=0.334, P=0.448). All patients underwent 18F-PSMA PET/CT within 4 weeks after biopsy. The maximum standardized uptake value(SUV max) and the minimum apparent diffusion coefficient(ADC min)were recorded, and the ratio of SUV max/ ADC minwere calculated. The correlation between the above parameters and ISUP grade group were analyzed.The diagnostic value of the parameters was evaluated by the receiver operating characteristic (ROC) curve. Results:The data of 49 patients were analyzed. The average ADC minwas (0.57±0.16)×10 -3 mm 2/s, with the average SUV max and SUV max/ADC min of 15.30±12.54 and (29.69±23.72)×10 3, respectively. Statistical differences were found in SUV max ( P=0.012) and SUV max/ADC min ( P=0.002) between the high- and low-grade groups, while ADC min ( P=0.411) showed no statistical differences between the two groups. Significant positive correlations were found between SUV max(r=0.501, P<0.001), SUV max/ADC min (r=0.527, P<0.001) and ISUP grade group, respectively. There was a negative correlation between ADC min and ISUP grade group (r=-0.296, P=0.039). SUV max/ADC min was the best index to distinguish high-grade group from low-grade group prostate cancer with the area under the curve(AUC) of 0.749. In contrast, the AUC of SUV maxand ADC min were 0.731 and 0.615, respectively. The diagnostic sensitivity and specificity of SUV max/ADC min were 73.3% and 85.3%, respectively, with a critical value of 37.23×10 3. Conclusion:The combination use of 18F-PSMA PET/CT and mpMRI could improve the diagnostic efficiency for prostate cancer, compared to either modality alone. The ratio of SUV max/ADC min has a positive correlation with ISUP grade group, and is a promising index for distinguishing the high-grade prostate cancer from low-grade cancer.

Objective:To analyze the clinicopathological features of positive surgical margins (PSM) after radical prostatectomy and to explore its associated factors.Method:A retrospective analysis was conducted on 274 patients who underwent radical prostatectomy in Beijing Hospital from June 2018 to June 2021. The margins of these specimens of radical prostatectomy were directly inked with black ink. According to the margin status (tumor present versus not), the patients were divided into PSM and negative surgical margin (NSM) groups. The clinicopathological characteristics were compared between two groups, including age, preoperative prostate specific antigen (PSA), number of tumors, tumor′s location, postoperative pathological Gleason score, tumor burden and postoperative pathological staging.Results:Among the 274 cases, 114 showed PSM, and 160 showed NSM. PSM accounted for 41.6% of the cases. The mean age was 68.3 years, while the PSM group′s mean age was 68.0 years, and that of the NSM group was 68.6 years, with no statistical significance between groups ( P>0.05). The mean preoperative PSA was 15.8 μg/L in all patients, 21.5 μg/L in the PSM group and 11.3 μg/L in NSM group. PSA in the PSM group was statistically higher than that in the NSM group ( P<0.001). The PSA level (10 μg/L, 10-20 μg/L, and >20 μg/L) was associated with the PSM rate (31.1%, 48.7%, and 69.4%). Regarding tumor numbers, 118 cases had a single focus, including 40 cases with PSM (33.9%). In the 156 cases of multiple foci, 74 cases had a PSM (47.4%). There were statistically more PSM cases in the cases with multi-focal disease ( P<0.05). Tumors were seen in the transit zone of 44 cases, while 107 cases showed tumors in the peripheral zone, and 123 cases in the whole zone. The PSM rate was 27.3% (12/44), 40.2% (43/107), and 48.0% (59/123) by tumor location, respectively, but the difference among groups was not statistically significant ( P>0.05). The postoperative Gleason scores were 3+3=6 in 51 cases, 3+4=7 in 98 cases, 4+3=7 in 81 cases, and ≥8 in 44 cases, with PSM rates of 19.6% (10/51), 38.8% (38/98), 45.7% (37/81) and 65.9% (29/44), respectively ( P<0.001 for rate differences). The tumor burden was <30% in 157 cases, 30%-60% in 91 cases, and>60% in 26 cases, with PSM rate of 21.0% (33/157), 65.9% (60/91) and 80.8% (21/26), respectively ( P<0.001 for rate differences). Moreover, there were 181 cases of pathological stage T2 (PSM rate, 29.3%) and 93 cases of pathological stage T3 (PSM rate, 65.6%), with statistical difference in PSM rates ( P<0.001). The multivariable logistic regression analysis indicated that preoperative PSA >20 μg/L, postoperative Gleason score ≥8, high tumor burden and pathological stags were different between the PSM and NSM groups ( P<0.05). Conclusions:The PSM of radical prostatectomy is closely related to the preoperative PSA level, the number of lesions, postoperative Gleason score, tumor burden and pathological stage. Preoperative PSA level >20 μg/L, postoperative Gleason score ≥8, high tumor burden and pathological stage are independent predictors for PSM.

Objective:To explore the diagnostic value of 18F-prostate specific membrane antigen (PSMA) PET/CT PRIMAY score combined with multiparameter MRI (mpMRI) PI-RADS score for clinically significant prostate cancer (CsPCa). Methods:The data of 63 patients with prostate cancer who underwent radical prostatectomy at Beijing Hospital from January 2019 to December 2023 were retrospectively analyzed. The median age was 70 (64, 75) years old with prostate-specific antigen (PSA) level of 8.46 (5.40, 14.80) ng/ml. All patients underwent 18F-PSMA PET/CT and mpMRI examination before surgery, and pathological large sections of prostate specimens were made after surgery. The prostate lesions were diagnosed and located by two radiologists and one pathologist respectively. Lesions with Gleason scores (GS)≥3+ 4 from the surgical pathology were diagnosed with CsPCa, and lesions with negative or GS=6 were diagnosed with non-CsPCa. The PSMA PET/CT images were evaluated using the PRIMARY study criteria (5-level PRlMARY score): no pattern (score of 1), diffuse transition zone or central zone(not focal) (score of 2), focal transition zone(score of 3), focal peripheral zone(score of 4), or an SUV max of at least 12 (score of 5). The degree of uptake of imaging agent in prostate lesions was semi-quantitatively evaluated using lesion-to-background ratios (LBR) of SUV max. MpMRI was evaluated according to the Prostate Imaging Reporting and Data System (PI-RADS) version 2.1. The patients were divided into CsPCa group and non-CsPCa group based on patients and lesions. Mann-Whitney U test and chi-square test were used to compare the differences between groups. Multivariate logistic regression analysis was performed to determine the independent predictive factors of CsPCa. Receiver operator characteristic (ROC) curve was used to determine the optimal diagnostic threshold for each independent predictor. Predictive models were constructed for PRIMARY score, PI-RADS score, and their combined application, and the diagnostic performance of each model for CsPCa was compared. Results:Of all 63 patients, there were 54 cases in CsPCa group (85.7%) and 9 cases in non-CsPCa group (14.3%).There was significant difference between CsPCa group and non-CsPCa group in the serum PSA level [9.64 (6.1, 15.3) ng/ml vs. 5.6 (4.6, 7.6) ng/ml]( P<0.05). There was no statistically significant difference in age [71 (64, 75) years vs. 65 (63, 69) years], and number of lesions [2 (1, 2) vs. 2 (1, 3)] (all P>0.05). Of all 109 lesions, there were 81 lesions in CsPCa group(including 49 lesions with Gleason score = 3+ 4, 16 lesions with Gleason score=4+ 3, 14 lesions with Gleason score = 8, and 2 lesions with Gleason score>8) and 28 lesions in non-CsPCa group(including 14 lesions with Gleason score = 3+ 3 and 14 with benign prostate lesions). There was significant difference between CsPCa group and non-CsPCa group in PRIMARY score [4 (3, 5) vs. 2 (1, 4)], LBR [2.69 (2.08, 4.48) vs. 1.89 (1.45, 2.48)], PI-RADS score [4 (3, 5) vs. 2 (2, 3)] (all P<0.05). There was no statistically significant difference in the lesion distribution including the number of lesions located in the transition zone [15(18.5%) vs. 8(28.6%)] and in the peripheral zone[66(81.5%) vs. 20(71.4%)]( P>0.05). Multivariate logistic regression analysis indicated that PRIMARY score ( OR=2.134, 95% CI 1.429-3.187) and PI-RADS score ( OR=2.689, 95% CI 1.618-4.469) were independent predictors of CsPCa (both P<0.01). ROC curves analysis revealed that the cut-off value for diagnosing CsPCa was both 3 for PRIMARY score and PI-RADS score. The accuracy for PRIMARY score, PI-RADS score, and their combined complication in diagnosing CsPCa was 72%, 67%, and 83%, respectively. The sensitivity was 72%, 63%, and 91%, and the specificity was 75%, 79%, and 57%, respectively. The positive predictive value was 89%, 89%, and 86%, and the negative predictive value was 48%, 42%, and 70%, respectively. The area under the curve of the PRIMARY score, PI-RADS score, and their combined complication of the ROC curve for CsPCa were 0.733 (95% CI 0.624-0.842), 0.708 (95% CI 0.599-0.817), and 0.743 (95% CI 0.623-0.862), respectively. The diagnostic efficacy of their combined complication was higher than PRIMARY score or PI-RADS score alone (both P<0.01). Conclusions:Both the 18F-PSMA PET/CT PRIMAY score and the mpMRI PI-RADS score have good diagnostic value for CsPCa. The combined application of the two imaging parameters can improve the accuracy, sensitivity, and negative predictive value, which have a higher diagnostic efficiency of CsPCa.

Objective:To analyze the prognosis of patients with positive resection margin after radical prostatectomy, as well as the prostate-specific antigen (PSA)level and risk factors for PSA progression.Methods:A retrospective analysis was performed on the data of 141 patients with pathologically diagnosed prostate cancer who underwent RP from May 2012 to August 2020 in Beijing Hospital. The mean age was (67.4±6.7)years, the preoperative median PSA was 9.6 (1.4-152.8) ng/ ml and the median follow-up time was 56 months. Postoperative pathology was T 2 stage 74 (52.5%), T 3 stage 63 (44.7%), T 4 stage 4 (2.8%). Biochemical recurrence after radical resection was defined as PSA rose to more than 0.2 ng/ml and showed an upward trend after two consecutive follow-ups. In this study, serum PSA ≥ 0.1 ng/ml without biochemical recurrence after radical operation was defined as PSA progression. The PSA level, risk factors of PSA progression and prognosis of patients with positive resection margin were analyzed. Univariate and multivariate Cox regression analysis was used to analyze the correlation between age, preoperative PSA level, pathological stage (pT), ISUP classification, surgical approach, lymph node dissection, single/multiple positive margins and PSA progression. Results:The median follow-up of 141 patients was 52 months(1-104 months). There were 69 (48.9%) patients in the PSA progression group and 72 (51.1%) patients in the non PSA progression group. In the PSA progression group, 13 (18.8%) patients did not receive treatment and 8 (61.5%) patients had biochemical recurrence. 4 (5.8%) patients received radiotherapy alone, and 2 (50.0%) patients had biochemical recurrence. 52 (75.4%) patients received endocrine therapy or endocrine therapy combined with radiotherapy, and 5 (9.6%) patients developed castration resistance. Multivariate Cox regression analysis showed preoperative PSA ( HR=1.015, 95% CI 1.005-1.025, P =0.004), ISUP grade and group ( HR=1.351, 95% CI 1.091-1.673, P =0.006), surgical method ( HR=2.233, 95% CI 1.141-4.370, P =0.019) was correlated with PSA progression. Conclusions:The incidence of surgical positive margin is high after RP. Nearly half of the patients with surgical positive margin developed a PSA progression status. Preoperative PSA, ISUP grade group, and the surgical approach are risk factors for PSA progression in patients with positive surgical margins. Patients with these risk factors should be monitored more closely and treated more aggressively.

Objective:To investigate the practicality and safety of performing a radical prostatectomy(RP)shortly after the diagnosis of prostate cancer using a combination of prostate targeted biopsy and intraoperative frozen section.Methods:Prospective enrollment was conducted for patients suspected of having prostate cancer based on abnormal prostate specific antigen(PSA)levels.The inclusion criteria for the study were as follows: patients aged 80 years or younger with an ECOG score of 1 or lower.Prior to biopsy, patients underwent both prostate magnetic resonance imaging(MRI)and prostate specific membrane antigen positron emission tomography/computed tomography(PSMA PET/CT)to determine the likelihood of prostate cancer with clinical stages within T 2-3aN 0M 0.In order to be included in the study, patients must agree to receive RP after their prostate cancer diagnosis has been confirmed by biopsy.All enrolled patients underwent a targeted prostate biopsy, consisting of 1-2 cores.These specimens were then examined through frozen section analysis.For patients diagnosed with prostate cancer through intraoperative frozen section pathology, RP was immediately performed.In this study, transperineal prostate targeted+ systematic biopsy was utilized for patients with undiagnosed prostate cancer.Additionally, routine pathological examination of specimens was conducted.The study analyzed the baseline data, surgical conditions, pathological results, and follow-up information of patients in a descriptive manner. Results:Seven patients, ranging in age from 54 to 77 years with a mean age of 66.7 years, were enrolled in the study.Their mean PSA level was 12.668 μg/L, ranging from 4.359 to 22.195 μg/L.Of these patients, 4 had a PI-RADS score of 4 and 3 had a score of 5.The maximum diameter of the index lesion was 1.3 cm, ranging from 0.5 to 2.2 cm.PSMA PET/CT scores were 4 in 1 case and 5 in 6 cases.The index lesions detected by PSMA PET/CT were consistent with those detected by MRI, and the maximum standardized uptake value(SUVmax)was 15.7, ranging from 5.3 to 39.4.Prostate cancer was diagnosed through targeted biopsy and intraoperative frozen section pathology.Four cases had a Gleason score of 3+ 3=6, while one case had a Gleason score of 3+ 4=7, another had a score of 4+ 3=7, and the last had a score of 4+ 4=8.All patients underwent RP treatment immediately after the prostate cancer diagnosis.Only one patient had slight adhesion at the apex of the prostate, while the other six patients were evaluated by surgeons as having no obvious adhesion at the apex.All surgeries were completed successfully, with a mean operation time of 149.7(ranging from 108 to 255)minutes.After RP, whole mount pathology results indicated that all cases were prostate adenocarcinoma, with a Gleason score of 3+ 4=7 in four cases and 4+ 3=7 in three cases.The pathological stages were pT2 in three cases and pT3a in four cases, with five cases having negative surgical margins and two cases with positive surgical margins.During the study, all patients were monitored for a period of 5.4 months(ranging from 3 to 7 months)and no complications of Clavien Dino≥Ⅰ were observed.PSA levels were measured at 6 weeks and 3 months after surgery, with readings of 0.020 μg/L(ranging from 0 to 0.079 μg/L)and 0.016 μg/L(ranging from 0 to 0.087 μg/L), respectively.No hormonal therapy or radiotherapy was administered during this time.Four patients were able to recover from urinary continence.Conclusions:Based on a combination of MRI and PSMA PET/CT, it is both safe and feasible to promptly perform RP following the diagnosis of prostate cancer through targeted biopsy for index lesions, along with intraoperative frozen section.