ObjectiveTo generate cynomolgus and rhesus monkey models carrying the Booroola fecundity (FecB) variant of bone morphogenetic protein receptor type 1B (BMPR1B) gene (designated BMPR1B^FecB) using adenine base edit technology and to explore genetic improvement strategies for enhancing the reproductive performance of laboratory macaques. MethodsFour single-guide RNAs (sgRNAs) were designed to target the BMPR1B c.746A>G locus in cynomolgus and rhesus monkeys. The sgRNAs were combined with the ABE8e-SpRY base editor, and the resulting editing complexes were microinjected into zygotes of both macaque species. Following in vitro culture of the injected embryos, nested PCR amplification was performed to detect the base-editing efficiency at the target site to screen for highly efficient sgRNAs. Subsequently, normally developing gene-edited embryos were transferred into the oviducts of surrogates. Seven days after birth, ear skin or peripheral blood samples were collected from the newborn monkeys for genotyping and mosaicism analysis, and the growth, development, and health status of the gene-edited monkeys were monitored. ResultsAmong the four candidate sgRNAs, sgRNA4 was identified as achieving 100% editing efficiency at the c.746A>G site. A total of 17 live monkey offspring were obtained via embryo transfer, including 8 cynomolgus monkeys and 9 rhesus monkeys. Genotyping results showed that all 8 newborn cynomolgus monkeys carried the c.746A>G point mutation, with an editing efficiency of 100% (8/8), of which 37.50% (3/8) were mosaics. Among the 9 newborn rhesus monkeys, 8 individuals carried the c.746A>G mutation, with an editing efficiency of 88.89% (8/9), of which 75.00% (6/8) were mosaics. ConclusionThe ABE8e-SpRY base-editing system enables precise editing at the c.746A>G (p.Q249R) locus of the BMPR1B gene in both cynomolgus and rhesus monkey embryos, resulting in the successful establishment of laboratory monkey models carrying the BMPR1B^FecB variant. These models provide a valuable foundation for future studies of the regulatory role of the BMPR1B^FecB variant in reproductive performance at the non‑human primate level, as well as for exploring genetic improvement strategies to enhance reproductive performance in laboratory monkeys.

Objective:To investigate the effect of PIK3CA mutations on the tumor immune microenvironment in Luminal breast cancer and evaluate the potential of Alpelisib-loaded pH-responsive polymer micelles in modulating the tumor immune microenvironment.Methods:PIK3CA mutations in breast cancer were analyzed using bioinformatics tools.A mouse xenograft model of Luminal breast cancer harboring a PIK3CA mutation was established,and alterations in the tumor immune microenvironment were examined using mass cytometry(CyTOF).During the period from August 2004 to December 2008 in Tianjin Medical University Cancer Hospital,tissue biopsies of 62 Luminal breast cancer patients in the BRCA cohort were collected Study the relationship between PIK3CA mutations and tumor immune microenvironment at the organizational level.Alpelisib-loaded polymer micelles(Alpelisib@MSPM)were synthesized,characterized,and evaluated for thera-peutic efficacy in Luminal breast cancer with PIK3CA mutations.Results:PIK3CA is one of the most frequently mutated genes in breast can-cer,with the highest prevalence in Luminal subtypes.CyTOF analysis demonstrated that PIK3CA mutations contribute to a tumor immun-osuppressive microenvironment in xenografts.Multiplex fluorescence immunohistochemistry revealed that PIK3CA-mutated tumors exhib-ited more infiltration of myeloid-derived suppressor cells(MDSCs)and less infiltration of CD8? T cells.The synthesized Alpelisib-loaded pH-re-sponsive polymer micelles had an average size of approximately 127 nm.Treatment with Alpelisib and Alpelisib@MSPM reduced tumor growth in mice with PIK3CA-mutated Luminal breast cancer.Notably,the proportion of MDSCs decreased,whereas CD8? T cell infiltration in-creased significantly,with the more pronounced effect observed in the Alpelisib@MSPM treatment group.Conclusions:PIK3CA mutations drive the formation of a tumor immunosuppressive microenvironment in Luminal breast cancer.Targeted Alpelisib delivery via pH-respons-ive polymer micelles significantly enhances therapeutic efficacy in PIK3CA-mutated breast cancer.

Objective To explore the expression of core proteoglycans(Decorin)and osteoglycine(Mimecan)in cervical cancer and their relationship with the characteristics of clinical pathological traits and the prognosis of patients.Methods A total of 148 patients with cervical cancer who underwent surgical treatment in our hospital were selected.The expression levels of Decorin and Mimecan were determined by immunohistochemistry(IHC).The positive expression rates of Decorin and Mimecan in patients with different clinicopathological features were compared,and the correlation between the two was analyzed.Cox regression model was used to assess risk factors for death in patients with cervical cancer.Kaplan-Meier survival curve was used to compare the survival rates of patients with positive and negative Decorin and Mimecan expression.Results The positive expression levels of Decorin and Mimecan were both lower in cervical cancer tissue than those in adjacent tissue of cancer(20.0%vs.75.0%,28.3%vs.63.3%,all P＜0.05).The positive expression rates of Decorin and Mimecan were lower in patients with a tumor diameter≥2 cm,FIGO stage Ⅲ-Ⅳ,lymph node metastasis and low tumor differentiation degree compared to those with a tumor diameter＜2 cm,FIGO stage Ⅰ-Ⅱ,no lymph node metastasis and moderately to well tumor differentiation degree(P＜0.05).There was a positive correlation of Decorin and Mimecan in cervical cancer tissue(r=0.686,P＜0.01).Multivariate Cox regression analysis confirmed that negative expression of Decorin(HR=2.365,95%CI:1.236-4.525)and Mimecan(HR=2.191,95%CI:1.322-3.631),FIGO stage Ⅲ-Ⅳ(HR=1.747,95%CI:1.147-2.660)and low tumor differentiation degree(HR=1.577,95%CI:1.035-2.404)were independent risk factors for mortality in cervical cancer patients(P＜0.05).The 5-year survival rate of patients with positive Decorin and Mimecan expression was higher than those with negative expression(79.3%vs.51.3%,73.8%vs.50.0%),and the median survival time was prolonged(75 months vs.60 months,74 months vs.60 months).Conclusion The positive expression rates of Decorin and Mimecan are reduced in cervical cancer tissue and may serve as potential biomarkers for prognostic evaluation of cervical cancer.

Objective The relationship between serum uric acid(UA)levels and gait kinematics characteristics in patients with cerebral small vessel disease(CSVD)was investigated.Methods Retrospective analysis was conducted on patients with CSVD from outparient clinics of the Neurology and Rehabilitation Department of Zhongshan Hospital affiliated with Guangzhou University of Chinese Medicine from January 2023 to December 2023.The general information of patients were collected and the gait of patients was analyzed using three-dimensional gait analysis.Patients were then divided into mild gait disorder group(0-1 points),moderate gait disorder group(2-3 points),and severe gait dysfunction group(4-5 points)based on gait results.The total burden of CSVD imaging and serum results such as UA were collected.The relationship between UA level and CSVD gait disorders was analyzed.Results This study recruited 105 CSVD patients.Patients were divided into different groups based on the severity of their gait disorder including 40 in the mild group,49 in the moderate group,and 16 in the severe group.The blood uric acid level in the moderate group(358.43±13.44)μmol/L was higher than that in the mild group(336.00±12.48)μmol/L,and the blood uric acid level in the severe group(289.94±11.88)μmol/L was lower than that in the mild and moderate groups(P<0.05).The MoCA score in the severe gait disorder group(21.38±0.13)was lower than that in the mild and moderate groups(28.05±0.09 vs.25.22±0.10)(P<0.05).The step width of the CSVD severe load group was(13.26±2.80)cm compared to the light and moderate load groups[(11.22±1.70)cm vs.(11.65±2.70)cm]increased(P<0.05),and the left swing phase in the severe group(35.90%)decreased compared to the mild and moderate groups(38.50%vs.37.20%)(P<0.05).Spearman correlation analysis showed a negative correlation between UA levels and CMB(r=-0.20,P=0.04).Hyperuricemia was negatively correlated with brain atrophy(r=-0.20,P=0.04).In patients with mild to moderate gait disorders,there was a positive correlation between hyperuricemia and the total burden of gait disorders(r=0.25,P=0.02),and hyperuricemia and right gait speed(r=-0.22,P=0.04),Right stride(r=-0.29,P<0.01),Left step speed(r=-0.32,P<0.01),Left step frequency(r=-0.29,P<0.01),The left stride was negatively correlated(r=-0.26,P=0.01).Conclusion In CSVD patients with mild to moderate gait disorders,the levels of uric acid and hyperuricemia are positively correlated with the total burden of gait disorders.The gait disorders are mainly characterized by reduced bilateral pace,bilateral stride,and left step frequency.

Objective The purpose of this study was to provide a more effective method for researching the prevention and treatment of Graves'disease by comparing the effects of two plasmid vectors expressing the human thyrotropin receptor(TSHR)A subunit gene in inducing an animal model of Graves'disease via electroporation.Methods Plasmids pcDNA3.1-THSR A,and pTriEx1.1-THSR A expressing the TSHR A subunit were constructed and used to induce Graves'disease by intramuscular injection with immediate electroporation once every 3 weeks for a total of 4 times.Mice in the control group were injected with PBS.One week after the second electroporation,blood was collected to measure serum thyrotropin receptor antibody(TRAb).Three weeks following the last electroporation,echocardiography was performed on the mice.Mice were sacrificed 4 weeks after the last electroporation;blood,thyroid,and orbital tissues were collected;serum total thyroxine(TT4)was measured;and histological examination was performed.Results The average concentrations of serum TRAb in the pcDNA3.1-TSHR A group(n=15)and the pTriEx1.1-TSHR A group(n=13)were(6.9±2.0)U/L and(7.5±2.2)U/L,respectively.The latter was significantly higher than that in the control group(4.9±0.5)U/L(P=0.033).The average concentrations of serum TT4 in the pcDNA3.1-TSHR A group and pTriEx1.1-TSHR A group were(41.4±23.8)ng/mL and(63.2±53.7)ng/mL,respectively,both higher than that in the control group:(20.2±4.0)ng/mL(P＜0.01).Thyroid pathology showed thyroid follicular epithelial hyperplasia with T-cell infiltration in the model group.Echocardiography showed that the left ventricle mass in the pTriEx1.1-TSHR A group was higher than those in the control group(P=0.007)and pcDNA3.1-TSHR A group(P=0.012).Orbital pathology showed fibrotic changes in the extraocular muscles of mice in the model groups.Conclusions Both pcDNA3.1 and pTriEx1.1 expressing the TSHR A subunit were able to induce Graves'disease in mice by electroporation,and the efficiency of the two plasmids in inducing hyperthyroidism and Graves'ophthalmopathy was similar.The efficiency of pTriEx 1.1-TSHR A in inducing thyrotoxic heart disease was better than that of pcDNA3.1-TSHR A.

Tongue squamous cell carcinoma (TSCC) is a prevalent malignancy that afflicts the head and neck area and presents a high incidence of metastasis and invasion. Accurate diagnosis and effective treatment are essential for enhancing the quality of life and the survival rates of TSCC patients. The current treatment modalities for TSCC frequently suffer from a lack of specificity and efficacy. Nanoparticles with diagnostic and photothermal therapeutic properties may offer a new approach for the targeted therapy of TSCC. However, inadequate accumulation of photosensitizers at the tumor site diminishes the efficacy of photothermal therapy (PTT). This study modified gold nanodots (AuNDs) with the TSCC-targeting peptide HN-1 to improve the selectivity and therapeutic effects of PTT. The Au-HN-1 nanosystem effectively targeted the TSCC cells and was rapidly delivered to the tumor tissues compared to the AuNDs. The enhanced accumulation of photosensitizing agents at tumor sites achieved significant PTT effects in a mouse model of TSCC. Moreover, owing to its stable long-term fluorescence and high X-ray attenuation coefficient, the Au-HN-1 nanosystem can be used for fluorescence and computed tomography imaging of TSCC, rendering it useful for early tumor detection and accurate delineation of surgical margins. In conclusion, Au-HN-1 represents a promising nanomedicine for imaging-based diagnosis and targeted PTT of TSCC.
Tongue Neoplasms/diagnostic imaging* ; Carcinoma, Squamous Cell/diagnostic imaging* ; Animals ; Gold/chemistry* ; Mice ; Photothermal Therapy/methods* ; Tomography, X-Ray Computed ; Photosensitizing Agents ; Metal Nanoparticles ; Humans ; Cell Line, Tumor

Objective To explore the value of preoperative amide proton transfer(APT)imaging combined with diffusion kurtosis imaging(DKI)in predicting lymphovascular space invasion(LVSI)in cervical cancer.Methods Fifty-one patients with cervical cancer who underwent preoperative MRI examination and had complete postoperative pathology were retrospectively selected.Pelvic MRI scans were performed 1-2 weeks preoperatively,and the corresponding APT values,mean kurtosis(MK)and mean diffusivity(MD)values were obtained respectively,and the occurrence of LVSI was determined based on postoperative pathology results.The predictive effects of APT-and DKI-derived parameters alone or in combination on the LVSI status of cervical cancer were compared.Results Among 51 cases of cervical cancer,36 cases had pathologically confirmed LVSI and 15 cases did not had LVSI.The area under the curve(AUC)of the receiver operating characteristic(ROC)curve for the preoperative APT values,MK and MD values alone and in combination to predict the LVSI status of cervical cancer were 0.820,0.788,0.762,0.894,0.885,and 0.896,respectively.APT values combined with DKI-derived parameters predicted LVSI better than when they were used separately,in which APT values combined with MK and MD values predicted LVSI of cervical cancer with the largest AUC,sensitivity of 91.7％,specificity of 80.0％,and Youden's index of 0.717.Conclusion Preoperative APT imaging and DKI have important value in predicting LVSI of cervical cancer,and the combined application of the two can improve the prediction efficacy,which has certain clinical application value.

Background:The incidence of perianal fistulizing Crohn's disease is increasing and can seriously affect patients'quality of life.The efficacy of perianal surgery combined with biological agents for perianal fistulizing Crohn's disease remains to be clarified.Aims:To evaluate the clinical efficacy of perianal surgery combined with anti-tumor necrosis factor(TNF)-α agents in the treatment of perianal fistulizing Crohn's disease systematically.Methods:Cohort studies on perianal surgery combined with anti-TNF-α agents in the treatment of perianal fistulizing Crohn's disease were retrieved from PubMed,Web of Science,Embase,Cochrane Library,CNKI,Wanfang Data,and SinoMed.The retrieval time span was from the establishment of each database to October 30,2024.Literatures were enrolled according to the inclusion and exclusion criteria,and the data were extracted.Newcastle-Ottawa scale was used to evaluate the quality of literature.Meta-analysis was conducted by RevMan 5.4 software.Results:A total of 9 literatures involving 736 patients with perianal fistulizing Crohn's disease were enrolled.Meta-analysis showed that perianal surgery combined with anti-TNF-α agents shortened the healing time of anal fistula(WMD=-0.76,95%CI:-1.06--0.46;Z=4.91,P<0.000 01),decreased the recurrence rate of perianal fistula(OR=0.47,95%CI:0.26-0.84;Z=2.55,P=0.01),and reduced the perianal disease activity index(PDAI)score(WMD=-1.51,95%CI:-1.84--1.18;Z=8.91,P<0.000 01)when compared with perianal surgery alone.However,no significant differences in complete healing rate,partial healing rate,and reoperation rate were found between the two groups(all P>0.05).Conclusions:Perianal surgery combined with anti-TNF-α agents in the treatment of perianal fistulizing Crohn's disease demonstrates superior efficacy compared to perianal surgery alone in terms of shortening healing time,reducing recurrence rate and decreasing PDAI score.

Objective: To explore the changes of mechanosensitive channels Piezos (Piezo 1 and Piezo 2) in neurogenic bladder (NB) of young rats and their effects，so as to provide reference for clinical search of new therapeutic targets. Methods: A total of 30 female young SD rats were divided into 5 groups based on random number table method：sham operation group (sham)，2-week nerve transection group (NB-2W)，6-week nerve transection group (NB-6W)，2-week nerve transection + Piezos inhibitor group (NB-P-2W) and 6-week nerve transection + Piezos inhibitor group (NB-P-6W)，with 6 rats in each group.The NB models were constructed by transecting the L6 and S1 spinal nerves of young rats.The NB-2W and NB-6W groups were not intervened after modeling，while the NB-P-2W and NB-P-6W groups were intraperitoneally injected with Piezos inhibitor GsMTx4 (10 mg/kg) every 2 days after modeling.Bladder cystometry and ultrasound were performed after 2 and 6 weeks of transection.The expressions of Piezos and fibrosis-related indexes (Collagen Ⅰ and α-smooth muscle actin) were detected in bladder tissues. Results: The results of bladder cystometry showed that the basal bladder pressure in NB-2W group was significantly increased，while it was slightly decreased but was still higher in NB-6W group than in the sham group (P<0.05).Basal bladder pressure was lower in NB-P-2W group than in NB-2W group，but was higher than that in the sham group; basal bladder pressure was lower in NB-P-6W group than in NB-6W group，but higher than that in the sham group (P<0.05).Compared with the sham group，the NB-2W and NB-6W groups had firstly increased and then decreased maximum cystometric capacity (MCC) (P<0.05).Compared with NB-2W group，NB-P-2W group had lower bladder leakage point pressure (BLPP)，but higher MCC and bladder compliance (BC) (P<0.05).Compared with NB-6W group，NB-P-6W group had significantly lower BLPP but higher MCC and BC (P<0.05).HE and MASSON staining and ultrasound results showed that，with the extension of nerve transection time，bladder fibrosis gradually worsened，the bladder wall became rough and thickened，calculi were visible inside，and hydronephrosis gradually appeared; the degree of fibrosis in NB-P-2W and NB-P-6W groups was less than that in NB-2W and NB-6W groups，and no hydronephrosis was observed in the upper urinary tract.In addition，Western blotting and immunohistochemical results showed that NB-2W and NB-6W groups had significantly higher relative expression levels of Piezos，Collagen Ⅰ and α-SMA than the sham group (P<0.01)，while NB-P-2W and NB-P-6W groups had lower relative expression levels of Piezos,Collagen Ⅰ and α-SMA than NB-2W and NB-6W groups (P<0.01). Conclusion: The increased expressions of mechanosensitive channels Piezos in NB young rats may be involved in the progression of bladder fibrosis，but its mechanism needs further study.

Background:The incidence of perianal fistulizing Crohn's disease is increasing and can seriously affect patients'quality of life.The efficacy of perianal surgery combined with biological agents for perianal fistulizing Crohn's disease remains to be clarified.Aims:To evaluate the clinical efficacy of perianal surgery combined with anti-tumor necrosis factor(TNF)-α agents in the treatment of perianal fistulizing Crohn's disease systematically.Methods:Cohort studies on perianal surgery combined with anti-TNF-α agents in the treatment of perianal fistulizing Crohn's disease were retrieved from PubMed,Web of Science,Embase,Cochrane Library,CNKI,Wanfang Data,and SinoMed.The retrieval time span was from the establishment of each database to October 30,2024.Literatures were enrolled according to the inclusion and exclusion criteria,and the data were extracted.Newcastle-Ottawa scale was used to evaluate the quality of literature.Meta-analysis was conducted by RevMan 5.4 software.Results:A total of 9 literatures involving 736 patients with perianal fistulizing Crohn's disease were enrolled.Meta-analysis showed that perianal surgery combined with anti-TNF-α agents shortened the healing time of anal fistula(WMD=-0.76,95%CI:-1.06--0.46;Z=4.91,P<0.000 01),decreased the recurrence rate of perianal fistula(OR=0.47,95%CI:0.26-0.84;Z=2.55,P=0.01),and reduced the perianal disease activity index(PDAI)score(WMD=-1.51,95%CI:-1.84--1.18;Z=8.91,P<0.000 01)when compared with perianal surgery alone.However,no significant differences in complete healing rate,partial healing rate,and reoperation rate were found between the two groups(all P>0.05).Conclusions:Perianal surgery combined with anti-TNF-α agents in the treatment of perianal fistulizing Crohn's disease demonstrates superior efficacy compared to perianal surgery alone in terms of shortening healing time,reducing recurrence rate and decreasing PDAI score.