Objective : To construct triggering receptor expressed on myeloid cells 2(TREM2) gene knockout(TREM2-/-) mice using CRISPR/Cas9 technology, to breed TREM2-/- mice and to analyze the genotype of TREM2-/- mice. Methods : CRISPR/Cas9 technology was used to selectively knock out exon 2-3 regions of TREM2 gene to construct a TREM2-/- mouse model, and the genetic background of all mice was C57BL/6J. Polymerase chain reaction(PCR) was used to identify the genotype of mice. Quantitative real-time PCR(qPCR) and Western blot were used to detect the expression level of TREM2 in major tissues of mice, and the authenticity and scientific nature of PCR identification results were verified from mRNA level and protein level. According to the sgRNA sequence, the possible off-target sites were predicted on the CCTop website, and the tail DNA of mice was extracted and amplified by PCR and then Sanger sequencing was performed to detect whether there was off-target effect in TREM2-/- mice. Results : TREM2-/- mice were successfully constructed by CRISPR/Cas9 technology, and the mice were genotyped. The results of agarose gel electrophoresis showed that the mouse genotype with only 415 bp band amplified was wild type(WT), the mouse genotype of the 449 bp band amplified only was TREM2-/-, and the mouse genotypes amplified with 415 bp and 449 bp double bands were heterozygous. qPCR results showed that compared with WT mice, the mRNA expression of TREM2 in heart and brain tissues of TREM2-/- mice was down-regulated(P-/- mice was reduced(P-/- mice. Conclusion TREM2-/- mice are successfully constructed and bred, a reliable genotype identification method is established, the genetic stability of the mouse model is verified, which will provide an important genetic animal model for the study of TREM2 gene function.

OBJECTIVE To evaluate the mechanisms underlying the antidepressant effect of ZBH2012001,a novel serotonin and norepinephrine reuptake inhibitor(SNRI),in general and its ability to enhance monoaminergic transmission and suppress neuroinflammation in particular.METHODS① Male ICR mice were divided into vehicle(distilled water),duloxetine(DLX,10 or 20 mg·kg-1)and ZBH2012001(5,10 and 20 mg·kg-1)groups.One hour following ig administration,the antidepressant effect of ZBH2012001 was evaluated using the tail suspension test(TST)and forced swimming test(FST).② Radioligand binding assay was conducted to evaluate the affinity of ZBH2012001 for human serotonin transporters(hSERTs)and human norepinephrine transporters(hNETs).③ Mice were divided into vehicle(distilled water),DLX(10 or 20 mg·kg-1)and ZBH2012001(5,10 and 20 mg·kg-1)groups.One hour following drug administration,the 5-hydroxytryptophan(5-HTP)-induced head-twitch test or yohimbine-induced lethality test were performed to evaluate the effect of ZBH2012001 on the function of the 5-hydroxytryptamine(5-HT)and norepinephrine(NE)systems.④ Mice were divided into vehicle(distilled water+0.1%acetic acid),reserpine model(distilled water+reserpine 5 mg·kg-1),DLX(DLX 20 mg·kg-1+reserpine 5 mg·kg-1)and ZBH2012001(ZBH2012001 5,10 and 20 mg·kg-1+reserpine 5 mg·kg-1)groups.One hour following drug administration,reserpine was injected intraperitoneally to establish a monoamine-depletion model.The ptosis,akinesia,and hypothermia assays were performed to evaluate the effect of ZBH2012001 on the down-regulation of the reserpine-induced monoamine system.The TST in mice was used to evaluate the effect of ZBH2012001 on reserpine-induced depressive-like behavior while high-performance liquid chromatography with electrochemical detection(HPLC-ECD)was used to measure the levels of monoamines and their metabolites in the hippocampal tissue of reserpine-induced monoamine-depletion mice.ELISA was employed to detect the contents of tumor necrosis factor-alpha(TNF-α)and interleukin-6(IL-6)in the hippocampal tissue of reserpine-induced monoamine-depletion mice.Western blotting was used to assess the expressions of ionized calcium-binding adapter molecule-1(Iba-1)and nuclear factor-kappa B(NF-κB)in the hippocampal tissue of reserpine-induced monoamine-depletion mice.RESULTS ① Compared with the vehicle group,ZBH2012001(5,10 and 20 mg·kg-1)significantly reduced the immobility time both in the TST in mice(P<0.01,respectively),and ZBH2012001(20 mg·kg-1)and in the FST in mice(P<0.05).② ZBH2012001 competitively inhibited the binding of[3H]-imipramine to hSERTs and[3H]-nisoxetine to hNETs,with the half maximal inhibitory concentration(IC50)values of 84.95 and 712.90 nmol·L-1,respectively.③Com-pared with the vehicle group,ZBH2012001(10 and 20 mg·kg-1)significantly increased the head twitches induced by 5-HTP in mice(P<0.01,respectively)and increased the mortality rate in mice induced by yohimbine(P<0.05,P<0.01).④ In the reserpine-induced monoamine-depletion model in mice,compared with the vehicle group,mice in the reserpine model group exhibited ptosis,akinesia and hypothermia feature(P<0.01,respectively),significantly prolonged immobility time in the TST(P<0.01),significantly decreased the levels of NE,5-HT and dopamine(DA)(P<0.05,P<0.01),significantly increased the metabolic conversion rate of 5-HT and DA(P<0.01,respectively),significantly elevated levels of TNF-α and IL-6(P<0.05,respectively),and significantly increased expressions of Iba-1 and NF-κB(P<0.05,respectively)in the hippocampus.Compared with the model group,ZBH2012001(5,10 and 20 mg·kg-1)significantly antagonized ptosis and hypothermia behaviors induced by reserpine(P<0.01,respectively),ZBH2012001(10 and 20 mg·kg-1)significantly shortened the immobility time in reserpine-treated mice(P<0.05,P<0.01),ZBH2012001(20 mg·kg-1)significantly increased the levels of NE and 5-HT in the hippocampus of reserpine-treated mice(P<0.05,respectively),decreased the metabolic conversion rate of 5-HT(P<0.05),significantly reduced the contents of TNF-α and IL-6 in the hippocampus of reserpine-treated mice(P<0.05,respectively),ZBH2012001(5,10 and 20 mg·kg-1)significantly reduced the expression of Iba-1 protein in the hippocampus of reserpine-treated mice(P<0.01,respec-tively),and ZBH2012001(20 mg·kg-1)significantly reduced the expression of NF-κB protein in the hippocampus of reserpine-treated mice(P<0.05).CONCLUSION ZBH2012001 exerts its antidepres-sant effect through a dual mechanism involving monoamine enhancement and inflammation suppres-sion.

Polycystic kidney disease (PKD) is a hereditary kidney disease characterized by the formation of numerous cysts in the kidneys, which progressively impairs renal function over time. PKD is primarily divided into two types: autosomal dominant polycystic kidney disease (ADPKD) and autosomal recessive polycystic kidney disease (ARPKD), with ADPKD being more prevalent. Current treatments primarily focus on symptom relief and disease progression delay, lacking a curative approach. However, the development of gene editing technologies such as clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (CRISPR-Cas9) and adeno-associated virus (AAV) vectors has offered new therapeutic possibilities for ADPKD and ARPKD. These include approaches like antisense oligonucleotides (ASO), adenovirus-mediated gene knockdown, CRISPR- Cas9, Pkd1 gene enhancement therapy, and the use of induced pluripotent stem cells (iPSCs), which have shown potential efficacy in animal models and early clinical studies. Despite facing technological challenges, ethical and legal issues, and high costs, gene therapy presents an unprecedented hope for PKD treatment. Future interdisciplinary collaboration and international cooperation are essential for developing more effective treatment strategies for PKD patients.

Objective:To investigate the value of preoperative prediction of Ki-67 expression status in breast cancer based on multi-phase enhanced MRI combined with clinical imaging characteristics prediction model.Methods:This study was retrospective. A total of 213 breast cancer patients who underwent surgical treatment at Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College between June 2016 and May 2017 were enrolled. All patients were female, aged 24-78 (51±10) years, and underwent routine breast MRI within 2 weeks prior to surgery. According to the different Ki-67 expression of postoperative pathological results, patients were divided into high expression group (Ki-67≥20%, 153 cases) and low expression group (Ki-67<20%, 60 cases). The radiomic features of breast cancer lesions were extracted from phase 2 (CE-2) and phase 7 (CE-7) images of dynamic contrast enhanced (DCE)-MRI, and all cases were divided into training and test sets according to the ratio of 7∶3. The radiomic features were first selected using ANOVA and Wilcoxon signed-rank test, followed by the least absolute shrinkage and selection operator method regression model. The same method of parameters selection was applied to clinical information and conventional imaging features [including gland classification, degree of background parenchymal enhancement, multifocal/multicentric, lesion location, lesion morphology, lesion long diameter, lesion short diameter, T 2WI signal characteristics, diffusion-weighted imaging (DWI) signal characteristics, apparent diffusion coefficient (ADC) values, time-signal intensity curve type, and axillary lymph nodes larger than 1 cm in short axis]. Support vector machine (SVM) was then used to construct prediction models for Ki-67 high and low expression states. The predictive performance of the models were evaluated using receiver operating characteristic (ROC) curves and area under cueve(AUC). Results:Totally 1 029 radiomic features were extracted from CE-2 and CE-7 images, respectively, and 9 and 7 best features were obtained after selection, respectively. And combining the two sets of features for a total of 16 features constituted the CE-2+CE-7 image best features. Five valuable parameters including lesion location, lesion short diameter, DWI signal characteristics, ADC values, and axillary lymph nodes larger than 1 cm in short axis, were selected from all clinical image features. The SVM prediction models obtained from the radiomic features of CE-2 and CE-7 images had a high AUC in predicting Ki-67 expression status (>0.70) in both the training set and the test set. The models were constructed by combining the CE-2, CE-7, and CE-2+CE-7 radiomic features with clinical imaging features, respectively, and the corresponding model performance in predicting Ki-67 expression status was improved compared with the models obtained by using the CE-2, CE-7, and CE-2+CE-7 radiomic features alone. The SVM prediction model obtained from CE-2+CE-7 radiomic features combined with clinical imaging features had the best prediction performance, with AUC of 0.895, accuracy of 84.6%, sensitivity of 87.9%, and specificity of 76.2% for predicting Ki-67 expression status in the training set and AUC of 0.822, accuracy of 70.3%, sensitivity of 76.1%, and specificity of 55.6% in test sets.Conclusion:The SVM prediction model based on DCE-MRI radiomic features can effectively predict Ki-67 expression status, and the combination of radiomic features and clinical imaging features can further improve the model prediction performance.

Objective:To explore the diagnostic value of synthetic magnetic resonance imaging (syMRI) quantitative parameters for benign and malignant breast lesions.Methods:From September 2018 to March 2019, a total of 43 cases of breast lesions which were confirmed by surgery and pathology in Cancer Hospital, Chinese Academy of Medical Sciences were enrolled in this study. All patients underwent syMRI sequence scans before and after enhancement except for conventional T2WI, DWI, and enhancement scans. GE AW4.7 workstation was used to generate syMRI parameter maps (T1, T2, proton density mappings), and ITK-SNAP software was used to delineate the volume of interest. The T1, T2, PD values before and after dynamic contrast enhanced (DCE) were obtained, and the change values of each parameter were calculated. Meanwhile, the apparent diffusion coefficient (ADC) and time intensity curve (TIC) of the lesions were measured. The differences of each parameter value were compared between benign and malignant breast lesions, and the receiver operating characteristic (ROC) curve was used to analyze the diagnostic performance of each parameter.Results:Among the 43 enrolled cases, 13 were benign and 30 were malignant. Among the syMRI parameters, the pre-enhancement parameters including T1pre (median 1 663.07 ms), T2pre (median 103.33 ms), post-enhancement parameters ΔT1 (median 1 022.68 ms) and ΔT2 (median 27.67 ms) of benign group, significantly higher than those of the malignant group (the medians were 1 141.74, 92.53, 664.95, and 16.19 ms, respectively, P<0.05). The ADC value of the benign group (median 1.66×10 -3mm 2/s) was significantly higher than that of the malignant group (median 1.00×10 -3mm 2/s, P<0.05). The benign group included 6 cases of TIC curve type Ⅰ, 5 cases of type Ⅱ, and 2 cases of type Ⅲ. The malignant group included 2 cases of TIC curve type Ⅰ, 17 cases of type Ⅱ, and 11 cases of type Ⅲ. The difference between the two groups was statistically significant ( P<0.05). The area under the ROC curve (AUC) of T1pre before DCE was 0.869, higher than 0.806 of ADC and 0.697 of TIC. When the best cut-off value of 1 282.94 ms was chosen, the sensitivity and specificity of diagnosis were 76.9% and 93.3%, respectively. The combination of T1pre and T2pre can further improve the diagnostic performance (AUC=0.908). Conclusions:Among the syMRI quantitative parameters, T1pre, T2pre, ΔT1 and ΔT2 have good value for the differential diagnosis of benign and malignant breast lesions. T1pre has the best diagnostic performance, and the combination of T1pre and T2pre can further improve the diagnostic performance.

Objective:To explore the diagnostic value of synthetic magnetic resonance imaging (syMRI) quantitative parameters for benign and malignant breast lesions.Methods:From September 2018 to March 2019, a total of 43 cases of breast lesions which were confirmed by surgery and pathology in Cancer Hospital, Chinese Academy of Medical Sciences were enrolled in this study. All patients underwent syMRI sequence scans before and after enhancement except for conventional T2WI, DWI, and enhancement scans. GE AW4.7 workstation was used to generate syMRI parameter maps (T1, T2, proton density mappings), and ITK-SNAP software was used to delineate the volume of interest. The T1, T2, PD values before and after dynamic contrast enhanced (DCE) were obtained, and the change values of each parameter were calculated. Meanwhile, the apparent diffusion coefficient (ADC) and time intensity curve (TIC) of the lesions were measured. The differences of each parameter value were compared between benign and malignant breast lesions, and the receiver operating characteristic (ROC) curve was used to analyze the diagnostic performance of each parameter.Results:Among the 43 enrolled cases, 13 were benign and 30 were malignant. Among the syMRI parameters, the pre-enhancement parameters including T1pre (median 1 663.07 ms), T2pre (median 103.33 ms), post-enhancement parameters ΔT1 (median 1 022.68 ms) and ΔT2 (median 27.67 ms) of benign group, significantly higher than those of the malignant group (the medians were 1 141.74, 92.53, 664.95, and 16.19 ms, respectively, P<0.05). The ADC value of the benign group (median 1.66×10 -3mm 2/s) was significantly higher than that of the malignant group (median 1.00×10 -3mm 2/s, P<0.05). The benign group included 6 cases of TIC curve type Ⅰ, 5 cases of type Ⅱ, and 2 cases of type Ⅲ. The malignant group included 2 cases of TIC curve type Ⅰ, 17 cases of type Ⅱ, and 11 cases of type Ⅲ. The difference between the two groups was statistically significant ( P<0.05). The area under the ROC curve (AUC) of T1pre before DCE was 0.869, higher than 0.806 of ADC and 0.697 of TIC. When the best cut-off value of 1 282.94 ms was chosen, the sensitivity and specificity of diagnosis were 76.9% and 93.3%, respectively. The combination of T1pre and T2pre can further improve the diagnostic performance (AUC=0.908). Conclusions:Among the syMRI quantitative parameters, T1pre, T2pre, ΔT1 and ΔT2 have good value for the differential diagnosis of benign and malignant breast lesions. T1pre has the best diagnostic performance, and the combination of T1pre and T2pre can further improve the diagnostic performance.

Objective To investigate the differences of gemstone spectral curve and CT value of gastric cancer with different pathological types and differentiation degrees. Methods 91 cases of preoperative gemstone CT images with gastric cancer were collected, including 24 cases of mucinous carcinoma, 67 cases of non?mucinous carcinoma, 16 cases of signet ring cell carcinoma, 8 cases of mucinous adenocarcinoma, 32 cases of moderately differentiated adenocarcinoma and 35 cases of poorly differentiated adenocarcinoma. Gemstone CT spectral imaging was performed preoperatively, and the spectral curve of the lesion in venous phase was obtained by using GSI Viewer software, the slope of the curve was calculated, and 11 monoenergetic CT values of 40～140 keV (10 keV interval) were measured. The gemstone spectral curves and CT values of gastric cancer with different pathological types and differentiation degrees are compared. Results The curve slopes of non?mucinous carcinoma, signet ring cell carcinoma and poorly differentiated adenocarcinoma were －1.92±0.53,－1.73±0.37 and －2.14±0.54, respectively. The absolute values were higher than those of mucinous carcinoma (－1.45±0.54), mucinous adenocarcinoma (－0.90± 0.34) and moderately differentiated adenocarcinoma (－1.67±0.41), and the differences were all statistically significant (P＜0.05). There were significant differences in monoenergetic CT values between mucinous and non?mucinous carcinomas at 40?140 keV ( all P＜0.05). The former was lower than the latter in different degrees, and the lower the energy, the greater the difference was. There were significant differences in monoenergetic CT values between signet ring cell carcinoma and mucinous adenocarcinoma at 40?100 keV (all P＜0.05); monoenergetic CT values between poorly differentiated adenocarcinoma and moderately differentiated adenocarcinoma at 40?90 keV showed statistically significant differences ( P ＜ 0. 05 ). Conclusions Gastric cancer with different pathological types and differentiation degrees have their characteristic spectral curves in venous phase, and the monoenergetic CT values are significantly different at low energy. The spectral curve of gemstone CT may be helpful to evaluate the pathological type and differentiation degree of gastric cancer before operation.

Objective To investigate the differences of gemstone spectral curve and CT value of gastric cancer with different pathological types and differentiation degrees. Methods 91 cases of preoperative gemstone CT images with gastric cancer were collected, including 24 cases of mucinous carcinoma, 67 cases of non?mucinous carcinoma, 16 cases of signet ring cell carcinoma, 8 cases of mucinous adenocarcinoma, 32 cases of moderately differentiated adenocarcinoma and 35 cases of poorly differentiated adenocarcinoma. Gemstone CT spectral imaging was performed preoperatively, and the spectral curve of the lesion in venous phase was obtained by using GSI Viewer software, the slope of the curve was calculated, and 11 monoenergetic CT values of 40～140 keV (10 keV interval) were measured. The gemstone spectral curves and CT values of gastric cancer with different pathological types and differentiation degrees are compared. Results The curve slopes of non?mucinous carcinoma, signet ring cell carcinoma and poorly differentiated adenocarcinoma were －1.92±0.53,－1.73±0.37 and －2.14±0.54, respectively. The absolute values were higher than those of mucinous carcinoma (－1.45±0.54), mucinous adenocarcinoma (－0.90± 0.34) and moderately differentiated adenocarcinoma (－1.67±0.41), and the differences were all statistically significant (P＜0.05). There were significant differences in monoenergetic CT values between mucinous and non?mucinous carcinomas at 40?140 keV ( all P＜0.05). The former was lower than the latter in different degrees, and the lower the energy, the greater the difference was. There were significant differences in monoenergetic CT values between signet ring cell carcinoma and mucinous adenocarcinoma at 40?100 keV (all P＜0.05); monoenergetic CT values between poorly differentiated adenocarcinoma and moderately differentiated adenocarcinoma at 40?90 keV showed statistically significant differences ( P ＜ 0. 05 ). Conclusions Gastric cancer with different pathological types and differentiation degrees have their characteristic spectral curves in venous phase, and the monoenergetic CT values are significantly different at low energy. The spectral curve of gemstone CT may be helpful to evaluate the pathological type and differentiation degree of gastric cancer before operation.

Objective: To investigate the differences of gemstone spectral curve and CT value of gastric cancer with different pathological types and differentiation degrees. Methods: 91 cases of preoperative gemstone CT images with gastric cancer were collected, including 24 cases of mucinous carcinoma, 67 cases of non-mucinous carcinoma, 16 cases of signet ring cell carcinoma, 8 cases of mucinous adenocarcinoma, 32 cases of moderately differentiated adenocarcinoma and 35 cases of poorly differentiated adenocarcinoma. Gemstone CT spectral imaging was performed preoperatively, and the spectral curve of the lesion in venous phase was obtained by using GSI Viewer software, the slope of the curve was calculated, and 11 monoenergetic CT values of 40~140 keV (10 keV interval) were measured. The gemstone spectral curves and CT values of gastric cancer with different pathological types and differentiation degrees are compared. Results: The curve slopes of non-mucinous carcinoma, signet ring cell carcinoma and poorly differentiated adenocarcinoma were -1.92±0.53, -1.73±0.37 and -2.14±0.54, respectively. The absolute values were higher than those of mucinous carcinoma (-1.45±0.54), mucinous adenocarcinoma (-0.90±0.34) and moderately differentiated adenocarcinoma (-1.67±0.41), and the differences were all statistically significant (P<0.05). There were significant differences in monoenergetic CT values between mucinous and non-mucinous carcinomas at 40-140 keV (all P<0.05). The former was lower than the latter in different degrees, and the lower the energy, the greater the difference was. There were significant differences in monoenergetic CT values between signet ring cell carcinoma and mucinous adenocarcinoma at 40-100 keV (all P<0.05); monoenergetic CT values between poorly differentiated adenocarcinoma and moderately differentiated adenocarcinoma at 40-90 keV showed statistically significant differences (P<0.05). Conclusions Gastric cancer with different pathological types and differentiation degrees have their characteristic spectral curves in venous phase, and the monoenergetic CT values are significantly different at low energy. The spectral curve of gemstone CT may be helpful to evaluate the pathological type and differentiation degree of gastric cancer before operation.

Keratinized mucosa in oral cavity plays an important role in periodontal health. The defect of keratinized mucosa may increase the risks of complication of oral implant surgery and restoration. Graft of keratinized tissue and connective tissue are still the gold standard for treating keratinized mucosa defect now. The current research focus on how to modulate non-keratinized mucosa to highly-efficient and minimally-invasive keratinized mucosa. Keratinocytes are critical components of oral mucosa and its final differentiation into keratinized mucosa is controlled by the connective tissue microenvironment involving a variety of molecules and ions. To fully understand keratinized differentiation of keratinocyte, this review focuses on its influence factors and possible mechanisms under the differentiation.