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 establish quality standards for pooled cryoprecipitate based on measured data, addressing the issues of wide variability in coagulation factor content attributable to individual donor differences and imprecise clinical dosing with single-bag cryoprecipitate. Methods: A total of 120 bags of fresh frozen plasma of different ABO blood groups were enrolled. Single-bag cryoprecipitate was prepared using either the centrifugation method or the siphonage method. For each blood group and each preparation method, every five single units of the same blood group were pooled into one bag of pooled cryoprecipitate. Factor Ⅷ content, fibrinogen content, and volume were measured in source plasma, single-bag cryoprecipitate, and pooled cryoprecipitate. Recovery rates and coefficients of variation were calculated, and the effect of the pooling process on product homogeneity was analyzed. Results: The factor Ⅷ content, fibrinogen content, and volume of pooled cryoprecipitate were significantly higher than those of single-unit cryoprecipitate and were linearly correlated with the number of units pooled. After pooling, the coefficient of variation for factor Ⅷ content decreased from 34.9%-35.2% (single units) to 18.2%-20.7% (pools), and the coefficient of variation for fibrinogen content decreased from 27.0%-28.6% to 13.1%-13.4%. The pooling procedure had no significant effect on the recovery rates of factor Ⅷ or fibrinogen (P>0.05). Conclusion: For pooled cryoprecipitate prepared by pooling five single units, the proposed quality standards are: factor Ⅷ content≥400 IU/bag, fibrinogen content≥750 mg/bag, volume 200-250 mL/bag, and sterility test negative. These standards can provide a basis for large-scale preparation and clinical application of pooled cryoprecipitate.

Pneumocystis jiroveci(PJ)is an atypical conditional pathogenic fungus.PJ colonization is a potential risk factor for Pneumocystis jiroveci pneumonia(PJP).Active identification of PJ colonization or infection is conducive to rational use of antibiotics.Although serum G test combined with molecular detection technology,peripheral blood CD4+T level and independent characterization of trophozoites and cysts by transcriptome sequencing technology may support identifying PJ colonization and diagnosis of infection,unified standards have not yet been established.Based on the pathogenic characteristics and pathogenic mechanism of PJ,clinical characteristics and diagnostic methods of PJP,this paper reviews the progress of differential diagnosis of PJ infection and colonization,so as to provide more objective and comprehensive differential strategy for clinical diagnosis and treatment.

Central retinal vein occlusion(CRVO)is a retinal vascular disorder that significantly impairs vision, with its underlying mechanisms involving complex interactions across multiple biological systems. This article provides a systematic review of the pathological mechanisms associated with CRVO, emphasizing critical factors such as endothelial dysfunction, arteriosclerosis, thrombophilia, inflammation, and oxidative stress. The pathological mechanisms of CRVO are characterized by arteriosclerosis, which obstructs venous return through a dual mechanism involving mechanical compression and endothelin-1-mediated contraction; endothelial dysfunction, which exacerbates disturbances in blood flow; genetic and acquired coagulation abnormalities that disrupt hemostatic balance and promote thrombosis; and the synergistic effects of inflammation and oxidative stress that activate cytokines, thereby aggravating ischemia and vascular leakage. Innovatively, this review explores emerging mechanisms such as miRNA-mediated vascular regulation via exosomes, gut microbiota-retina crosstalk through the “gut-eye axis,” and systemic metabolic interactions that link local retinal lesions to broader dysregulation of CRVO. These insights underscore the importance of integrated eye-system interventions and provide a theoretical foundation for advancing early biomarker discovery, multitarget therapeutics, and personalized treatment paradigms. By bridging localized pathology and systemic mechanisms, this work promotes a transformative shift toward an integrative medicine model in the diagnosis and management of CRVO.

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.

Objective To investigate the effects of LINC01355 on the proliferation,apoptosis,and invasion of oral squamous cell car-cinoma(OSCC)cells via the miR-545-5p/forkhead box D1(FOXD1)signaling pathway.Methods Cal-27 cells were cultured in vitro and randomly separated into the control group,si-LINC01355(transfected with LINC01355 siRNA)group,pc-LINC01355(transfected with LINC01355 overexpression plasmid)group,si-NC+miR-545-5p-NC+pc-NC(transfected with LINC01355 siRNA negative control,miR-545-5p negative control and empty plasmid)group,and si-LINC01355+miR-545-5p inhibitor(transfected with LINC01355 siRNA and miR-545-5p inhibitor)group.After transfection,quantitative real-time PCR was applied to detect the expression of LINC01355,miR-545-5p,and FOXD1 in cells.The Cal-27 cells transfected into groups were then subcutaneously inoculated to construct nude mouse models of transplanted tumors in each group,and the growth of the transplanted tumors was measured.The CCK-8 method,flow cytom-etry,and Transwell assay were applied in order to detect cell proliferation,apoptosis,and invasion,respectively.Immunohistochemical staining was applied to detect the expression of epithelial-mesenchymal transition(EMT)related proteins Vimentin and E-cadherin in cells.Western blotting was applied to detect the expression of cell proliferation related proteins(proliferating cell nuclear antigen[PCNA],C-myc),apoptosis related proteins(cleaved cysteinyl aspartate-specific proteases-3[cleaved caspase-3],BCL-2-associated X protein[Bax]),and FOXD1 protein.A dual-luciferase reporter assay was used to identify the relationship between LINC01355 and the miR-545-5p/FOXD1 signaling pathway.Results Compared with the control group,the expression of LINC01355 and FOXD1 mRN A,proliferation activity,number of invasions,positive expression of Vimentin protein,expression of PCNA,C-myc and FOXD1 protein,and transplanted tumor volume reduced(All P＜0.05);the expression of miR-545-5p,apoptosis rate,cleaved caspase-3 and Bax protein expression,and positive expression of E-cadherin protein increased(All P＜0.05)in the si-LINC01355 group.The trend of changes in the various indica-tors of the pc-LINC01355 group was opposite to that of the si-LINC01355 group.Compared with the si-LINC01355 group,the expression of LINC01355 and FOXD1 mRNA,prolife ration activity,number of invasions,positive expression of Vimentin protein,expression of PCNA,C-myc and FOXD1 proteins,and the transplanted tumor volume in the si-LINC01355+miR-545-5p inhibitor group increased(All P＜0.05),whereas the expression of miR-545-5p,apoptosis rate,cleaved caspase-3 and Bax protein expression,and positive expression of E-cadherin protein decreased(All P＜0.05).LINC01355 was able to downregulate miR-545-5p expression in Cal-27 cells and miR-545-5p was able to downregulate FOXD1 expression.Conclusion LINC01355 promotes OSCC cell proliferation and invasion,and inhibits apoptosis by targeting the miR-545-5p/FOXD1 signaling pathway.

OBJECTIVE: To compare effectiveness of multiple metatarsal osteotomy versus first metatarsophalangeal arthrodesis in treating severe metatarsal adductus hallux valgus deformity. METHODS: A retrospective analysis was conducted on the clinical data of 25 patients with severe metatarsal adductus hallux valgus deformity admitted between June 2010 and May 2014 who met the selective criteria. Among them, 15 patients underwent multiple metatarsal osteotomy (osteotomy group), while 10 patients underwent first metatarsophalangeal arthrodesis (fusion group). There was no significant difference between groups ( P>0.05) in gender, age, disease duration, affected side, preoperative American Orthopaedic Foot and Ankle Society (AOFAS) score, visual analogue scale (VAS) score for pain, intermetatarsal angle (IMA), hallux valgus angle (HVA), or metatarsal adduction angle (MAA). The osteotomy group underwent fixation with screws and/or staples fixation, while the fusion group utilized anatomic fusion plates and trans-articular compression screws. The study compared the following outcome indicators between groups: operation time, pre- and post-operative differences (change values) in AOFAS scores, VAS scores, and radiographic parameters (HVA, MAA), osteotomy healing outcomes, and recurrence of hallux valgus deformity. RESULTS: Both surgical procedures were completed successfully. The operation time was significantly shorter in the fusion group than in the osteotomy group ( P<0.05). All patients were followed up 96-144 months (mean, 116 months). The follow-up time was (129.1±7.2) months in the osteotomy group and (104.4±8.0) months in the fusion group, with no significant difference between groups ( P>0.05). X-ray films revealed the radiographic union in two groups, and the fusion time was significantly shorter in the fusion group than in the osteotomy group ( P<0.05). At last follow-up, both groups demonstrated significant improvements in AOFAS and VAS scores compared to preoperative levels ( P<0.05). However, the differences in the change values of AOFAS and VAS scores between groups were not significant ( P>0.05). During follow-up, 3 cases (20%) of deformity recurrence occurred in the osteotomy group, while no recurrence was observed in the fusion group. There was no significant difference in the incidences of deformity recurrence between groups ( P>0.05). CONCLUSION For severe metatarsus adductus hallux valgus deformities, both multiple metatarsal osteotomy and first metatarsophalangeal arthrodesis can correct the deformity. The former preserves metatarsophalangeal joint mobility but demands high technical proficiency from the surgeon, involves relatively longer operation times, extended bone healing periods, and higher complication incidences. The latter procedure is relatively simpler, facilitates faster postoperative recovery, allows early weight-bearing, and yields more reliable outcomes, though it sacrifices first metatarsophalangeal joint mobility.
Humans ; Osteotomy/methods* ; Hallux Valgus/diagnostic imaging* ; Retrospective Studies ; Arthrodesis/instrumentation* ; Treatment Outcome ; Metatarsal Bones/diagnostic imaging* ; Metatarsophalangeal Joint/diagnostic imaging* ; Male ; Female ; Bone Screws ; Adult ; Middle Aged ; Bone Plates ; Pain Measurement

Objective:To investigate the effects of electroacupuncture stimulation on the expression of Nrf2 and GPX4 proteins in the hippocampal CA1 region of rats with middle cerebral artery occlusion(MCAO)and its neuroprotective mechanism.Methods:Forty-eight rats were divided into the normal group(Normal),the sham operation control group(Sham),the model group(MCAO)and the electroacupuncture group(EA)according to the random number table method.The right middle cerebral artery occlusion model was established in the MCAO and EA groups by the modified thread embolization method.Starting on day 1 post-modeling,the EA group received electroacupuncture stimulation at the"Baihui"point(GV20)and left"Neiguan"point(PC6)for 30 min daily over 14 d.The Normal group received no treatment,and the Sham group only underwent vascular isolation without inserting the suture.Neurological function of rats in each group were scored before and after modeling and before and after electroacupuncture stimulation using the Zea Longa scoring scale.The structure and arrangement of Nissl-positive cells in the hippocampal CA1 region were observed by Nissl staining.Western blot and immunohistochemical staining were used to determine the expression levels of Nrf2 and GPX4 proteins in the hippocampal CA1 region.Results:Compared with the Normal group,the Sham group showed no neurological deficits,abundant Nissl-positive cells,and no statistically significant differences in Nrf2 and GPX4 protein expression or positive cell counts.Compared with the Sham group,the MCAO group exhibited elevated Zea Longa score,reduced Nissl-positive cell counts,and decreased Nrf2 and GPX4 protein expression and positive cell counts(P＜0.05).In contrast,the EA group demonstrated lower Zea Longa scores,increased Nissl-positive cell counts,and elevated Nrf2 and GPX4 protein expression and positive cell counts compared to the MCAO group(P＜0.05).Conclusion:Electroacupuncture stimulation may protect against neuronal damage in the hippocampal CA1 region of MCAO rats,potentially through up-regulation of Nrf2 and GPX4 expression.

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: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.