Recent innovations in nanomaterials inspire abundant novel tumor-targeting CRISPR-based gene therapies. However, the therapeutic efficiency of traditional targeted nanotherapeutic strategies is limited by that the biomarkers vary in a spatiotemporal-dependent manner with tumor progression. Here, we propose a self-amplifying logic-gated gene editing strategy for gene/H₂O₂-mediated/starvation multimodal cancer therapy. In this approach, a hypoxia-degradable covalent-organic framework (COF) is synthesized to coat a-ZIF-8 in which glucose oxidase (GOx) and CRISPR system are packaged. To intensify intracellular redox dyshomeostasis, DNAzymes which can cleave catalase mRNA are loaded as well. When the nanosystem gets into the tumor, the weakly acidic and hypoxic microenvironment degrades the ZIF-8@COF to activate GOx, which amplifies intracellular H⁺ and hypoxia, accelerating the nanocarrier degradation to guarantee available CRISPR plasmid and GOx release in target cells. These tandem reactions deplete glucose and oxygen, leading to logic-gated-triggered gene editing as well as synergistic gene/H₂O₂-mediated/starvation therapy. Overall, this approach highlights the biocomputing-based CRISPR delivery and underscores the great potential of precise cancer therapy.

Objective:To validate the diagnostic performance of the Paris system for reporting urinary cytology (TPS).Methods:A total of 7 046 urine cytology samples from 3 402 patients collected in the Department of Pathology, Beijing Hospital, China from January 2020 to January 2022 were analyzed. 488 patients had a biopsy or resection specimen during the follow-up period of 6 months. The sensitivity, specificity, risk of malignancy (ROM) and risk of high-grade malignancy (ROHM) of the TPS were evaluated using histological diagnosis as the golden standard.Results:Among the 7 046 samples, high-grade urothelial carcinoma (HGUC) accounted for 5.7% (399/7 046), suspicious for high-grade urothelial carcinoma (SHGUC) for 3.2% (227/7 046), atypical urothelial cells (AUC) for 8.4% (593/7 046), and negative for high-grade urothelial carcinoma (NHGUC) for 72.9% (5 139/7 046) including low-grade urothelial neoplasm (LGUN) for 0.8% (59/7 046) and insufficient samples for 9.8% (688/7 046). 488 patients had a bladder biopsy or resection in the follow-up of six months, including 314 males and 174 females, aged 27 to 92 years (average, 66 years). The ROHM of TPS was 94.7% in HGUC, 83.3% in SHGUC, 41.3% in AUC and 18.8% in NHGUC. The sensitivity and specificity of urine cytology were 70.1% (169/241) and 97.0% (162/167), respectively. The negative predictive value of NHGUC was 69.2% (162/234).Conclusions:The study has shown that TPS classification has high sensitivity and specificity, high ROHM for HGUC and SHGUC, and high negative predictive value for NHGUC. The application of TPS reporting system can better interpret the clinical significance of cytology samples, improve the accuracy of urine cytopathology and ensure continuous diagnostic consistency.

Purpose To assess the feasibility of nuclear score combined with ultrasound features on indeterminate thyroid nodules in fine needle aspiration cytological diagnosis.Methods A consecutive cohort of 119 thyroid fine-needle aspiration(FNA)specimens with indeterminate diagnosis(TBSRTC cate-gory Ⅲ-Ⅴ)and available histopathologic follow-up was evalua-ted.Original sonographic images and cytological slides of each nodule were reviewed by two experienced sonographers and two experienced cytopathologists respectively,and evaluated jointly to obtain consensus.The identification of the optimal cut-off points of simplified nuclear score and ultrasound features score for the diagnosis of malignancy or low-risk neoplasm were evalu-ated using the receiver operating characteristic(ROC)curves and the assessment of the area under the ROC curve(AUC).The specificity,sensitivity,positive predictive value(PPV)and negative predictive value(NPV)of nuclear score and US score were evaluated from crosstabs based on Cut-off points and signif-icance were calculated.The diagnostic accuracy of simplified nuclear score combined with ultrasound features was estimated by Logistic regression and ROC curve analysis.Results Nucle-ar grooves,intra-nuclear inclusions and chromatin clearing were more common in malignancy/low-risk neoplasms(P = 0.001,0.013 and 0.001 respectively).A Cut-off point of≥2 for the simplified nuclear score was sensitive for defining malignancy/low-risk neoplasm,and the sensitivity was 99.1%.Nodule height greater than width,absent halo sign,irregular margin and absent intranodular vascularization were more common in malig-nancy/low-risk neoplasms(P = 0.003,0.007,0.003 and 0.004 respectively).When the Cut-off point was set as≥5,the PPV and specificity for defining malignancy/low-risk neoplasm were 95.9%and 78.6%respectively.ROC curve analysis showed that the diagnostic accuracy of nuclear score combined with US score(0.873,95%CI = 0.777-0.970)was higher than that of nuclear score alone(0.817,95%CI = 0.687-0.948,P =0.092)or US score alone(0.758,95%CI 0.637-0.879,P =0.047).But no significant difference was detec-ted between the combined score and the nuclear score.Conclu-sion The addition of simplified nuclear score and US score to FNA cytology can increase the diagnostic accuracy of cytology in indeterminate categories.

Objective:To observe the expression of adenosine kinase (ADK) in the hippocampus of patients with refractory epilepsy, and to explore the role of ADK in the pathogenesis of refractory epilepsy.Methods:Thirteen patients with intractable epilepsy who underwent surgical resection of hippocampal tissue at the First Affiliated Hospital of Harbin Medical University were collected as the epilepsy group; At the same time, 4 cases of relatively normal temporal lobe brain tissue from patients with traumatic brain injury undergoing debridement surgery (without previous history of epileptic seizures) were collected, and these 4 patients served as the control group. The expression of ADK in two groups of specimens was detected at the tissue, gene, and protein levels using methods such as dual fluorescence immunohistochemistry, real-time quantitative polymerase chain reaction (RT Real time PCR), and Western blotting.Results:In the human brain, ADK was mainly expressed in the nucleus of astrocytes. Through histological observation, ADK was weakly expressed in normal brain tissue, while there is significant proliferation of glial cells and excessive expression of ADK in the brain tissue of patients with refractory epilepsy. The percentage of ADK positive glial cells in the epilepsy group was (53.90±17.59)%, and the control group was (23.82±4.18)%, with a statistically significant difference ( P<0.01). At the genetic level, using RT Real time PCR, it was found that the expression level of ADK mRNA in the epilepsy group was higher than that in the control group, with a 2 -△△Cp of 13.36, which was 13.36 times higher than that in the control group. At the protein level, the expression of ADK protein in the epilepsy group was found to be higher than that in the control group using protein immunoblotting ( P<0.01). Conclusions:ADK is weakly expressed in the nucleus of astrocytes in normal human brain tissue. In the brain tissue of patients with refractory epilepsy, astrocytes significantly proliferate and there is excessive expression of ADK. ADK may play an important role in the occurrence and development of refractory epilepsy in humans.

Objective:To study the significance of induction chemotherapy and subsequent comprehensive therapy for overall survival rate (OS) and larynx dysfunction-free survival rate (LDFS) in patients with advanced hypopharyngeal carcinoma.Methods:Patients who met the inclusion criteria with the diagnoses of advanced hypopharyngeal carcinoma between 2011 and 2017 received 2 or 3 cycles of TPF regimen induction chemotherapy. Patients who attained complete response (CR) received radical chemotherapy. Patients who attained partial response (PR) and the reduction of tumor volume was more than 70% were defined as large PR and received concurrent chemoradiotherapy. When the tumor volume reduction of PR patients was less than 70%, they were defined as small PR. (CR+large PR) group was defined as effective group. Patients who did not reach CR and large PR were defined as uneffective group and underwent radical surgery and received adjuvant radiotherapy as appropriate after the surgery. The end points of the study were OS, progression-free survival (PFS) and LDFS. Chi-square (χ 2) test was used for correlation analysis. Survival analysis was performed by the Kaplan-Meier method with a Log-rank test. Cox proportional hazards model was used for univariate and multivariate survival analysis. Results:A total of 260 patients were enrolled in the study. The follow-up period ranged from 5 to 83 months, with an average of 24.7 months. The 3-year and 5-year OS rate was 46.0% and 32.6%, respectively. The 3-year and 5-year PFS rate was 41.0% and 26.6%, respectively. The 3-year and 5-year LDFS rate was 37.9% and 24.8%, respectively. Poor outcome of induction chemotherapy, advanced N stage, strong positive Ki-67 immunohistochemistry (all P<0.001) were negative prognostic factors. The advanced clinical stage was positively related to the poor outcome of induction chemotherapy ( P=0.015). There was no significant difference in OS and PFS between the large PR group and the small PR group (all P>0.005). Conclusion:TPF regimen induction chemotherapy and subsequent comprehensive therapy for patients with advanced hypopharyngeal carcinoma may improve the quality of life of patients, with high OS rate and LDFS rate.

Objective To investigate the current status of urinary incontinence (UI) at the third trimester of pregnancy among primiparas in Guangdong Province.Methods The convenience sampling method was adopted to conduct the retrospective analysis on 714 primiparas in 3 hospitals of Guangdong Province from June 2015 to March 2016.International Consultation on Incontinence Questionnaire Urinary Incontinence Short-Form Chinese was used to investigate the occurrence status of UI at the third trimester of pregnancy among primiparas.And its influencing factors were analyzed.Results Among 714 primiparas,192 cases (26.9％) developed UI during the third trimester of pregnancy,in which stress UI accounted for 67.2％ (129/192).The prepregnancy BMI and abortion had statistical difference between.the UI patients and non-UI patients (P＜0.05).The multivariate Logistic regression results showed that pre-pregnancy BMI was correlated with UI at the trimester stage of pregnancy (OR =1.077,P＜0.05).Conclusion Pre-pregnancy BMI might be an independent risk factor of UI occurrence at the third trimester of pregnancy.

Objective: To elucidate the clinicopathological characters and prognostic factors of invasive micropapillary carcinoma of the breast (IMPC) by compared with invasive ductal carcinoma, not otherwise specified of the breast (IDC). Methods: The retrospective study was performed with female patients who had undergone curative resection for breast cancer without neoadjuvant chemotherapy from June 2008 to April 2016 in Breast Center of Beijing Hospital. Forty-seven mixed or pure IMPC patients and 93 pure IDC patients(admitted in the same center from October 2008 to January 2016 ) were matched for tumor stage, nodal status and age. Follow-up was done every 3 to 6 months postoperatively. The deadline was July 31, 2016. The curves of disease free survival and overall survival were drawn by the Kaplan-Meier method, and survival rates were compared by means of the Log-rank test. Potential prognostic variables that were identified on univariate analysis were analyzed with Cox′s proportional hazards regression model for multivariate analysis. The χ² test or Fisher′s exact test was used to compare distributions across 2 groups and the Mann-Whitney U test or t test was used to analyze the medians or means of 2 groups. Results: With exact matches, the rates of lymphovascular invasion (LVI) (29.8% vs. 12.9%, χ²=5.885, P=0.015)and histological grade 3 (40.4% vs. 21.5%, χ²=-2.690, P=0.007) were both significantly higher in patients with IMPC than that in IDC group, but the survival between the two pathological types were not significantly different (all P>0.05). The percent of IMPC component didn′t influence the clinicopathologic characters (all P >0.05), but a significantly longer median disease free survival (χ²=11.731, P=0.001) when the patients had more than 50% of IMPC component was found. Conclusions Higher rates of LVI and histological grade 3 were found in IMPC than that in IDC, but the survival was comparable between the two groups. A longer DFS occurred in patients with IMPC component more than 50%.

Objective: To determine the frequency and extent of left ventricular amyloid deposition in patients aged over 85 years with heart failure and preserved ejection fraction (HFpEF). Methods: A total of 43 patients aged 85 to 100 years old were enrolled in this study based on the autopsy database of Beijing Hospital from February 1, 2003 to October 31, 2016. The frequency and extent of left ventricular amyloid deposition and myocardial fibrosis were determined in left ventricular specimens from patients with antemortem diagnosis of HFpEF without clinically apparent amyloid (n=28) and from control subjects (n=15) post Congo red staining and Masson's trichrome staining. Kappa test was used to evaluate the consistency of the myocardial amyloidosis and fibrosis. Results: The heart weight of the patients in HFpEF group and in control group were similar((452.7±107.7)g vs. (415.0±70.8)g, t=-1.218, P=0.23)). Positive Congo-red staining was found in 24 examples (24/28) in HFpEF group and 5 examples (5/15) in the control group; severe amyloid deposition was found in 7 examples (7/28) in HFpEF group, but not in the control group. Amyloid deposition was more severe in HFpEF group than in control group (χ²=12.205, P<0.01). Masson's trichrome staining evidenced moderate to severe fibrosis in 19 cases (19/28) in HFpEF group and 8 cases (8/15) in control group (χ²=1.019, P=0.35). A consistent evaluation of the degree of myocardial fibrosis and the degree of myocardial amyloid deposition in all selected participants was performed and results showed that these two parameters were not consistent (Kappa value=0.2, P=0.820). Conclusion Amyloid deposition is common in the elderly patients with heart failure and preserved ejection fraction, suggesting that myocardial amyloidosis may be related to the development of HFpEF. There is no significant correlation between myocardial amyloidosis and myocardial fibrosis in this cohort.

Objective: To analyze the differences of clinicopathological characters and prognostic factors between invasive micropapillary carcinoma of the breast (IMPC) and invasive ductal carcinoma (IDC) not otherwise specified of the breast. Methods: Patients who were treated from June 2008 to April 2016 in Breast Center of Beijing Hospital were retrospectively analyzed to evaluate the differences between IMPC (n=59) and IDC (n=1 080). Follow-up was done every 3 to 6 months postoperatively with a deadline of July 31, 2016. The curves of disease free survival (DFS) and overall survival (OS) were drawn by Kaplan-Meier method, and survival rates were compared by means of the Log-rank test. Potential prognostic variables which were identified on univariate analysis were analyzed with Cox′s proportional hazards regression model for multivariate analysis. Results: More lymph nodes were involved in IMPC group (χ²=12.168, P=0.007) which led to more later stage in this group (χ²=8.950, P=0.011). IMPC group displayed a significantly increased rate of lymphovascular invasion (LVI) compared to IDC group (χ²=13.511, P = 0.001). The expression rate of estrogen receptor (ER) and progesterone receptor (PR) was higher in IMPC group than that in IDC group (89.8% vs. 76.3% and 88.1% vs. 70.7%, respectively, χ²=5.786, 8.332, all P<0.05). In multivariate analysis performed with the variables found significant in univariate analysis, the only variable found significantly affecting DFS of IMPC group was the T stage (T1-2 and T3-4, OR=5.217, 95%CI: 1.401 to 19.430, P=0.014), while in IDC group, pathological stage (stage Ⅰ to Ⅱ and stage Ⅲ to Ⅳ, OR=1.870, 95% CI: 1.262 to 2.771, P=0.002), lymph node positive ratio (LNR) (OR=2.222, 95%CI: 1.561 to 3.162, P=0.000), PR (OR=1.856, 95%CI: 1.118 to 3.082, P=0.017), and age (<50 years old and ≥50 years old, OR=0.695, 95%CI: 0.488 to 0.989, P=0.043) were prognostic factors. There were two variables found significantly affecting OS of IMPC group, which were T stage (OR=3.713, 95%CI: 1.539 to 8.959, P=0.004) and LNR (OR=2.850, 95%CI: 1.033 to 7.862, P=0.043). While in IDC group, LNR was the only variable found significantly affecting OS (OR=2.129, 95%CI: 1.324 to 3.425, P=0.002). Compared with IDC, the patients with IMPC were more likely to have local or regional recurrence (P=0.006). Although the median DFS interval was longer in IDC group (χ²=9.739, P=0.002), the median OS interval was comparable between the two groups (χ²=0.787, P=0.375). Conclusion Although IMPC has lymphotropic feature, tendency of LVI and local or regional recurrence, it has an OS which is comparable with IDC.

Objective: To study the cytomorphologic features and determine whether pancreatic neuroendocrine tumors (PanNET) sampled by fine-needle aspiration (FNA) can be accurately graded based on the Ki-67 index when compared to surgical samples. Methods: Corresponding intraoperative (19 cases) or endoscopic ultrasound-guided (3 cases) FNA cytology and surgical tissue specimens were obtained from 22 tumors, which were reviewed and stained for Ki-67 proliferation marker. The cytological samples included more than 200 tumor cells. Samples were graded by scoring the Ki-67 positive index in accordance with the 2010 WHO criteria. The grading scores assigned to the FNA cytology samples were compared with the scores assigned to the corresponding histological samples. Concordance was achieved by using 5% (instead of 2%) as a cut-off value for defining G2 tumors. One cytological sample included less than 500 tumor cells was excluded in the concordance calculation. Results: The cytological smears consisted of uniform, monotonous and isolated cells, loose cellular aggregates and rosette-like formations. Some tumor cells clustered around segments of capillaries. The cells demonstrated distinct cytoplasmic and nuclear features. Mitoses and necrosis were rarely seen. When traditional 2% Ki-67 index cut-off value were used to classify G2 tumors, the majority (86.4%, κ=0.812, P<0.01) of FNA cytology samples and corresponding surgical tissue specimens demonstrated concordance. When a 5% cut-off value was adopted, the concordance rate was 95.5% (21/22, κ=1.000, P<0.01). Similar concordance rates between the cytological and histological grades were achieved with threshold value of cytological assessment material set at more than 500 or 200 cells. Conclusions The cytological Ki-67 index in adequate material (>200 tumor cells) is useful in grading pancreatic neuroendocrine tumors, and a cut-off value of 5% showed better predictive value compared with that of 2%. Accurate grading of PanNET is critical for predicting tumor biology, patient prognosis, and making informed decisions regarding patient management and treatment.