BACKGROUND:With the extensive development of national fitness activities,nighttime exercise has gradually become a social trend.However,what effects nighttime exercise will have on sleep,the characterization of sleeping changes and the mechanisms of action are not clear. OBJECTIVE:To analyze the characterization and mechanisms of nighttime exercise on sleep. METHODS:We searched the China National Knowledge Infrastructure(CNKI),PubMed database,and Web of Science database with the search terms of"sport,exercise,physical activity,physical exercise,workout,sleep,asleep,daily routine"in Chinese,and"nocturnality,night,nighttime,exercise,sport,physical activity,sleep,slumber,dorm"in English for literature on how nighttime exercise affects sleep and its mechanism of action.Finally,55 articles were included. RESULTS AND CONCLUSION:Sleep consists of two phases:rapid eye movement and non-rapid eye movement.Sleep regulation relies on the close coordination of neural circuits and molecules in different brain regions,while the homeostatic components of the organism and circadian rhythms are also important factors in the regulation of sleep.The effects of exercise on sleep have evolved from an early focus on the central nervous system to the addition of somatic physiology,and now multiple dimensions have been investigated,revealing multifaceted effects of exercise on sleep,including subjective sleep quality,sleep continuity,and sleep quantity and sleep structure.Nighttime exercise-affected sleep is subjected to the constraints of exercise conditions and exercisers and shows certain complexity,but the heterogeneity,time-dependence,and instability characteristics of nighttime exercise-affected sleep can provide some guidance for nighttime exercisers in minimizing the impact on sleep.Mechanisms by which nighttime exercise affects sleep are related to the reduction of melatonin secretion,the increase of core body temperature,and energy expenditure.People with high sensitivity of their own sleep to nighttime exercise should avoid nighttime exercise as much as possible,and those with low sensitivity should also pay attention to the fact that the time of nighttime exercise should have a long interval with sleeping time,and the intensity should not be too high.Some measures can be adopted to safeguard sleep after exercise,such as energy replenishing after exercise,and reduced use of electronic devices.

Objective:To investigate the accuracy and feasibility of applying rapid immunohistochemistry(IHC) with CK5/6 antibodies in prostate biopsy tissues to assist frozen pathology diagnosis.Methods:The data of 41 patients who underwent prostate puncture and frozen tissue rapid IHC with CK5/6 antibody in Beijing Hospital from October 2022 to April 2023 were retrospectively analyzed. The median age of the patients was 76 (69, 79) years old, and the median PSA value was 12.37 (7.07, 26.17) ng/ml.The Prostate Imaging Reporting and Data System (PI-RADS) scores of the target lesions were all ≥3. The PI-RADS score of 9 patients(21.95%) was 3, 15 (36.59%) was 4, and 17(41.46%) was 5. The median diameter of the lesions in the MRI examination was 1.40 (1.09, 2.20) cm.Fourteen lesions (34.14%) were located in the migratory zone, 23 (56.10%) were located in the peripheral zone, and 4 (9.76%) involved both peripheral and migratory zone lesions. Transperineal cognitive fusion targeted combined systematic biopsy was used, and intraoperatively, 1 additional needle was taken from each of the target and non-target areas for frozen pathology section, and hematoxylin eosin(HE) staining and rapid IHC staining with CK5/6 antibody was performed, then the frozen remaining tissue was HE staining and CK5/6 IHC staining. Data such as HE and rapid IHC results of frozen pathology sections and HE and IHC results of routine sections of the frozen remaining tissues, International Society of Urological Pathology (ISUP) grading groupings(GG), and actual diagnostic results of targeted combined systematic puncture were recorded. Using the routine IHC results of the same needle tissue as the gold standard, the sensitivity and positive predictive value of applying rapid IHC frozen pathology to diagnose prostate cancer and the accuracy of its pathological GG were analyzed.Results:Among the 41 patients, a total of 35 cases were diagnosed with prostate cancer(PCa) by HE staining in frozen section of target tissue, with a positivity rate of 85.37%(35/41). Among these, there were 17 cases (48.57%) in ISUP GG 1, 8 cases (22.86%) in GG 2, 4 cases (11.43%) in GG 3, and 6 cases (17.14%) in GG 4 to 5. In addition, a total of 35 cases were diagnosed with PCa by HE staining in frozen remaining section of target tissue, with a positivity rate of 85.37%(35/41). Among these, there were 17 cases (48.57%) in ISUP GG 1, 8 cases (22.86%) in GG 2, 4 cases (11.43%) in GG 3, and 6 cases (17.14%) in GG 4 to 5. The results of rapid IHC of target tissue: 35 cases were negative for CK5/6 expression and 6 cases were positive. The results of routine IHC of target tissue: 35 cases were negative for CK5/6 expression and 6 cases were positive. The results of rapid IHC of non-target tissue: 12 cases were negative for CK5/6 expression and 29 cases were positive. The results of routine IHC of non-target tissue: 12 cases were negative for CK5/6 expression and 29 cases were positive. Thirty-five cases of target tissue rapid IHC diagnosis of PCa were in complete agreement with routine IHC diagnosis, with a false-positive rate of 0, a sensitivity of 100.00% (35/35) and a positive predictive value of 100.00% (35/35). Twelve cases of non-target tissue rapid IHC diagnosis of PCa were in complete agreement with routine IHC diagnosis, with a false-positive rate of 0, a sensitivity of 100.00% (35/35), and a positive predictive value of 100.00% (12/12).Conclusions:The preliminarily study results confirmed that the application of rapid IHC with CK5/6 antibodies in prostate biopsy tissues assisted frozen pathology diagnosis with high accuracy, but the reliability of rapid IHC technology in assisting frozen pathological diagnosis during puncture surgery still needs further validation through large sample size prospective studies.

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 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.

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 investigate the feasibility and reliability of the frozen section during targeted prostate biopsy.Methods:The clinical and pathological information of patients who received cognitive fusion transperineal targeted plus systematic biopsy and frozen section of 1-2 core targeted biopsy were consecutively collected and retrospectively studied. The median age was 70 (ranging 64-78) years, with the median prostate-specific antigen (PSA) level of 11.00 (ranging 6.63-16.52) ng/ml and the median prostate volume of 35.72 (ranging 22.59-47.71) ml. All patients received bi-parametric magnetic resonance imaging (bp-MRI) and have Prostate Imaging Reporting and Data System (PI-RADS) 3 or higher lesions diagnosed on bp-MRI. The suspected lesions would be taken by targeted biopsy of which one or two cores would be sent to prepare for the frozen sections. Then a cognitive fusion targeted and systematic biopsy covering the above targeted zones would be routinely administered under a transperineal approach as a standard protocol. The total time used for diagnosis of the frozen sections, the pathological diagnosis and the International Society of Urological Pathology (ISUP) grade groups (GG) would be recorded. The sensitivity, the positive predictive value, and the accuracy on grade groups would be analyzed, using the pathological diagnosis based on standard sections from the same targeted lesion.Results:A total of 29 patients were included in this study. Accordingly, 29 suspected lesions were identified on bp-MRI. A total of 20 lesions were finally diagnosed of PCa on frozen section, with the detection rate of 69.0%. Of those, 9(45.0%) cases were ISUP GG 1 diseases, 5(25.0%) cases were GG 2 diseases, 1(5.0%) case was GG 3 disease, and 5(25.0%) cases were GG 4-5 diseases. A total of 22 lesions were diagnosed with PCa on standard sections of cores from the same targeted lesions, with the detection rate of 75.9%. Of those, 6(27.3%) cases were GG 1 disease, 11(50.0%) cases were GG 2 diseases, 1(4.5) case was GG 3 disease, and 4(18.2%) cases were GG 4-5 diseases. The sensitivity and the positive predictive value of frozen section were 90.9% and 100%, respectively. No false positive diagnosis was made by frozen section. Compared to diagnosis from frozen sections, the GG diagnosed from final standard sections were found to upgrade and downgrade in 2 and 2 cases, respectively. The accuracy rate on GG of frozen sections was 80%. The time used for the diagnosis of frozen sections was (11±2) minutes. The histology quality control of four specimens was dissatisfactory. Two were due to tissue loss and deformation during sampling, and the other two were due to cytoclasis during low-temperature transferring.Conclusion:It is feasible and reliable to make a pathological diagnosis from frozen section of prostate targeted biopsy.

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 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 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 describe the 40-years trend for the mortality of colorectal cancer (CRC) in Shanghai and to estimate the effect of age, period, and birth cohort with Age-Period-Cohort (APC) model. Methods: Data on tumor-releated death from 1975 Janurary 1 to 2014 December 31 was derived from the Yangpu District of Shanghai Center for Diseases Prevention and Control tumor registration system. Colonrectal cancer cases (C18.2-C18.9 and C20 in ICD10) were selected for analyses. Crude mortality, age-adjusted mortality, and Average Annual Percent Changes (AAPCs) were calculated for colon cancer and rectal cancer. The difference of AAPCs between male/female and different age groups were tested. An APC model (reference cohort and period were 1900 and 1975, respectively) was constructed to estimate the age-effect, period-effect, and cohort-effect on the colorectal cancer death. Results: During 1975-2014, 6 725 cases died of colorectal cancer (the cased of colon and rectal cancer were 3 684 and 3 041, respectively). The crude mortality and age-adjusted mortality of colon cancer was 8.83/100 000 and 6.76/100 000, respectively. The crude mortality and age-adjusted mortality of rectal cancer were 7.32/100 000 and 5.67/100 000, respectively. For population in Yangpu District, the crude mortality and age-adjusted mortality of colon cancer increased with time, and the crude mortality of rectal cancer increased with time (P<0.001). AAPC of the crude mortality rate (5.6%) and age-adjusted mortality rate (2.3%) of colon cancer were higher than those in rectal cancer (3.0% and -0.3%), respectively (both P values <0.001). AAPC of the crude mortality rate (males vs. females was 6.2% vs. 5.0%, P<0.05) and age-adjusted mortality rate (males vs. females was 2.7% vs. 1.7%, P<0.05) of colon cancer were higher in males than in females. APC model indicted that CRC-related death increased with age. During 1901 to 1941, the RR values of cohort effects for colon and rectal cancer death were 1.09-5.57 and from 1.04-2.28, respectively; During 1946 to 1991, the RR values of cohort effects for colon cancer and rectal cancer were 5.51-4.32 and 2.16-0.89. Conclusion From 1975 to 2014, the mortality of CRC in Yangpu District increased gradually, and colon cancer mortality in males increased faster than that in females. The risk of death from colorectal cancer in the 1946-1991 birth cohort declined.