Objective:To investigate the current status of clinical practice of optical surface guided radiation therapy (SGRT) technology in China.Methods:A survey questionnaire was designed based on a similar investigation conducted by the European Society for Radiotherapy and Oncology in collaboration with the American Association of Physicists in Medicine on SGRT. The questionnaire covered aspects such as the installation, implementation, commissioning, quality assurance, clinical application, challenges, and cost considerations of SGRT systems. An online questionnaire was distributed to 49 institutions in China that have installed or are in the process of installing SGRT systems. Data were summarized and analyzed using Excel and SPSS 29 software.Results:Among the 49 institutions, 96% had at least one SGRT system. In terms of commissioning, quality assurance and implementation, it was mainly operated by physicists (94%) and technicians (82%), the cycle of test items for quality assurance was only achieved by the highest percentage of units with end-to-end test items for the annual inspection (50%). Eighty-six percent of the institutions used phantoms provided by suppliers, and 53% followed supplier recommendations or guidelines. For the installation of the first SGRT system, 37% of the institutions reported that initial staff training required more than 48 hours, while 73% found the training content easy to understand. Regarding the clinical application of SGRT technology, the majority of the institutions (53%) had used it for 1-3 years, with breast radiotherapy being the most commonly used treatment site. The primary scenario of SGRT application was intra-fraction motion monitoring / patient monitoring (69%). Furthermore, 47% of the institutions combined SGRT with open-face masks, and 71% used visual feedback devices for breath-hold or free-breathing gating. In terms of treatment thresholds, the median thresholds for monitoring and positioning were the same for breast, abdominopelvic (non- stereotactic body radiation therapy), and head-and-neck (non-brain stereotactic radiosurgery) treatments but varied for other sites.Conclusions:Although SGRT technology requires a relatively long initial training period, it is generally well accepted in terms of training and operation. Clinically, SGRT has been widely applied in breast radiotherapy, playing a crucial role in patient monitoring and intra-fraction motion management. However, most institutions have had limited clinical experience with the technology, highlighting the need for continuous technical supervision and improvement. The establishment of standardized protocols is necessary to ensure broader clinical adoption and long-term effectiveness.

Objective:To discuss the effect of DEAD-box RNA helicase 39A(DDX39A)gene silencing on the proliferation,migration and invasion of the esophageal cancer TE-1 cells,and to clarify its possible mechanism.Methods:For bioinformatics analysis,GSE63941,GSE77861,GSE20347,and GSE16153 chip data were downloaded from the GEO database.The esophagel cancer-related data were selected from the TCGA Database.R software was used to analyze the differentially expressed genes.STRING Database was used to construct the protein-protein interaction(PPI)network.Identification of key genes of high relevance was achieved using the MCODE plugin in Cytoscape.The expression of key genes in normal esophageal tissue and esophageal cancer tissue were analyzed with the GEPIA 2 database.Kaplan-Meier Plotter was used to perform survived analysis and plotting for the screened key genes.Cytological experiments were carried out on esophageal cancer TE-1 cells,and small interfering RNA(siRNA)technology was used to silence the expression of DDX39A gene.The TE-1 cells in the logarithmic growth phase were selected and divided into blank(MOCK)group,negative control(si-NC)group,and silencing(si-DDX39A)group.Real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods were used to detect the expression levels of DDX39A mRNA and protein in the TE-1 cells in various groups;CCK-8 assay was conducted to detect the proliferation activity of cells in various groups,and the cell scratch assay was used to measure the migration rate of cells in various groups;Transwell chamber assay was used to detect the number of invasion cells in various groups;Western blotting method was used to detect the expression levels of β-catenin,glycogen synthase kinase-3β(GSK3β),phosphorylated glycogen synthase kinase-3β(p-GSK3β),c-MYC,Cyclin D1 and nuclear β-catenin proteins in the cells in various groups.Results:Analyses using TCGA database combined with the GEO Database yielded a total of 56 differentially expressed genes.MCODE plugin in Cytoscape software identified 41 key genes of high relevance;DDX39A was screened by analyzing 41 genes through the GEPIA 2 and Kaplan-Meier plotter Databases.The results of RT-qPCR and Western blotting methods showed that compared with si-NC group,the expression levels of DDX39A mRNA and protein in the cells in si-DDX39A group were decreased(P＜0.05).The CCK-8 results showed that the proliferation activity of the cells in si-DDX39A group was lower than that in si-NC group(P＜0.05).The cell scratch assay results showed that the cell migration rate in si-DDX39A group after 24 h was lower than that in si-NC group(P＜0.05).The results of Transwell chamber assay showed that the number of invasion cells in si-DDX39A group was lower than that in si-NC group(P＜0.05).Compared with si-NC group,the expression levels of β-catenin,p-GSK3β,c-MYC,Cyclin D1,and nuclear β-catenin in the TE-1 cells in si-DDX39A-1 group and si-DDX39A-3 group were decreased(P＜0.01),but the expression levels of GSK3β protein had no significant differences(P＞0.05).Conclusion:Silencing of DDX39A gene could inhibit the proliferation,migration and invasion of TE-1 cells,and the mechanism may be related to the regulation of Wnt/β-catenin signaling pathway.

Objective:To investigate the clinical and genetic characteristics of a Henan Han family with pontine autosomal dominant microangiopathy and leukoencephalopathy (PADMAL), aiming to enhance understanding of this disease.Methods:The proband was first admitted to the Department of Neurology, Henan Provincial People′s Hospital, Fuwai Central China Cardiovascular Hospital in December 2019 due to cerebral infarction and unilateral limb numbness and weakness. Detailed medical history collection, pedigree mapping, whole-exome sequencing screening, and Sanger sequencing validation were performed for the proband and family members. The patients′ clinical manifestations, imaging features, neuropsychological scale assessment results, and pathological changes were summarized, and genetic analysis was conducted on the gene variant site. Relevant literature was reviewed to summarize the characteristics of PADMAL.Results:The proband was a 47-year-old female, with 3 generations of family members affected, including 7 patients, 3 of whom had died. The clinical features of the patients were similar, with the first stroke occurring around the age of 40, without vascular risk factors such as hypertension or diabetes. The main clinical manifestation was unilateral limb numbness and weakness. The proband and her niece sought medical attention due to stroke symptoms. Brain magnetic resonance imaging revealed acute infarct lesions located in the pons, accompanied by multiple oval infarct foci (the "raisin bread sign") and white matter hyperintensity changes. Genetic testing showed that 4 patients carried a heterozygous c. *34GT mutation in the 3′-untranslated region (3′-UTR) of the COL4A1 gene, while the other 4 unaffected family members did not carry this variant, consistent with genotype- phenotype co-segregation in the family. Conclusions:PADMAL is an extremely rare monogenic cerebral small vessel disease caused by pathogenic variants in the 3′-UTR of the COL4A1 gene. The "raisin bread sign" in the pons is a relatively specific imaging feature that distinguishes it from other cerebral small vessel diseases. For patients with this sign, genetic testing for PADMAL should be considered.

Objective To evaluate the impact of slow-freezing process on human ovarian tissue with the standard cryopreservation-thawing protocol of Fertility Protection Center of Beijing Obstetrics and Gynecology Hospital,Capital Medical University.Methods Ovarian tissues of 12 patients were divided into fresh ovarian tissue group(fresh group)and freezing-thawing ovarian tissue group(F-T group).The freezing-thawing protocol was the standard protocol in our center.The number and activity of follicle were examined with Hematoxylin-eosin(HE)staining and calcein-AM(calcein acetoxymethylester)staining,and the proliferation and apoptosis was evaluated with the immunohistochemical staining of Ki-67 and caspase-3.The expressions of apoptosis-related proteins such as caspase-3,bax and FasL between the two groups were compared with Western blotting.Results There were no statistically significant differences in follicle counting and follicle activity in ovarian tissues pre-and post-freezing-thawing(P>0.05),and the positive rate of Ki-67 in ovarian tissues after freezing-thawing was significantly lower than that in fresh ovarian tissues(P<0.05),and there was no statistically significant difference in the positive rate of caspase-3 between the two groups(P>0.05).The expression of caspase-3 protein in ovarian tissues after freezing-thawing was significantly higher than that in fresh ovarian tissues(P<0.05),while the expressions of other apoptosis-related proteins such as bax and FasL were not significantly different(P>0.05).Conclusion The standard cryopreservation-thawing regimen in our center can effectively maintain the follicle number,morphology,and activity in ovarian tissues.After freezing and thawing,the cell proliferation level is decreased.The expression of apoptosis-related proteins such as bax and FasL are not increased,and the expression of caspase-3 is relatively increased.These results suggest our freezing-thawing regimen is good for human ovarian tissue.

Female infertility is recognized as a global public health issue by the World Health Organization.Female fertility remodeling includes ovarian function reconstruction and uterus/endometrium reconstruction,etc.It is emerging as a hot technology since it is ready to prepare autologous platelet-rich concentrate and it is safer and more acceptable in autologous application.It plays an important role in regenerative medicine,and it is currently widely applied in maxillofacial and plastic surgery,dermatology and other clinical practices.This article mainly reviews the progresses of the application of autologous platelet-rich concentrate in female fertility remodeling.

Objective:To construct a full-length infectious clone of enterovirus D68（EV-D68）expressing enhanced green fluorescent protein（EGFP）by reverse genetics in order to provide an efficient tool for studying the biological characteristics and screening antiviral drugs for EV-D68.Methods:Gene synthesis and overlap PCR techniques were used to construct the full-length clone plasmid pUC57-EV-D68 of EV-D68. The full-length viral sequence was then transferred into the pCAGGS plasmid to obtain the pCAGGS-EV-D68 plasmid. The EGFP gene was amplified and inserted into the pCAGGS-EV-D68 plasmid to construct the pCAGGS-EGFP-EV-D68 plasmid. Then，the two constructed plasmids were transfected into human rhabdomyosarcoma（RD）cells to rescue recombinant viruses RV-EV-D68 and RV-EGFP-EV-D68. The rescued viruses were identified using PCR，Western blot，and immunofluorescence techniques. The antiviral effect of doxycycline was evaluated using the rescued RV-EGFP-EV-D68. Statistical analysis was performed using the two independent samples t-test. Results:The recombinant virus RV-EGFP-EV-D68 capable of expressing EGFP was successfully rescued. Even after 15 serial passages，the virus retained EGFP expression with no significant difference in viral titers compared to the parental virus，indicating its stable passage in RD cells. Besides，the rescued strains exhibited similar replication characteristics to the parental virus. While at 24 and 36 h after infection，the titers of the rescued strains were significantly lower than that of the parental strain（both P<0.05）. This study demonstrated that doxycycline significantly reduced the fluorescence intensity of RV-EGFP-EV-D68-infected RD cells（ P<0.01）. Meanwhile，a negative correlation was observed between the doxycycline concentration and the fluorescence intensity，indicating that the rescued virus could be used for antiviral drug evaluation. Conclusions:This study successfully constructs an infectious clone of EV-D68 expressing EGFP. The rescued recombinant virus RV-EGFP-EV-D68 has been verified to be applicable for the evaluation of antiviral drugs.

Objective To summarize the diagnosis and treatment of neuronal intranuclear inclusion disease （NIID） with peripheral neuropathy as the initial symptom in a Han Chinese family， and to improve the clinical awareness of this disease through a literature review. Methods A pedigree investigation was performed for a patient with NIID who was admitted to the outpatient service of Department of Neurology， Henan Provincial People’s Hospital， in January 2023， and the clinical and imaging features of the family members were summarized. Various techniques， such as second-generation whole-exome sequencing， third-generation whole-genome sequencing， and PCR capillary electrophoresis， were used for genetic analysis， and a family lineage map was plotted. A literature review was performed to summarize the features of cases with peripheral neuropathy as the initial symptom. Results There were 3 generations and 16 members in this family， among whom 5 had clinical manifestations， 8 underwent blood sampling， and 3 were found to have a GCC repeat expansion mutation in the 5' non-coding region of the NOTCH2NLC gene. The proband was aged 39 years， with the initial symptom of weakness in the extremities， and cranial MRI did not show the characteristic signal of NIID， while electromyography suggested multiple peripheral nerve injuries in the extremities. Genetic testing showed that the proband， his aunt， and his cousin all had a GGC repeat expansion mutation in the NOTCH2NLC gene， with a number of more than 60 repeats （154，144，and 148，respectively）. Conclusion In hereditary peripheral neuropathy， the possibility of NIID should be considered in case of negative results for common pathogenic genes. In addition， third-generation sequencing has a marked diagnostic value in repeat expansion mutation-related disorders.

The diagnosis of hematological disorders is currently established from the combined results of different tests, including those assessing morphology (M), immunophenotype (I), cytogenetics (C), and molecular biology (M) (collectively known as the MICM classification). In this workflow, most of the results are interpreted manually (i.e., by a human, without automation), which is expertise-dependent, labor-intensive, time-consuming, and with inherent interobserver variability. Also, with advances in instruments and technologies, the data is gaining higher dimensionality and throughput, making additional challenges for manual analysis. Recently, artificial intelligence (AI) has emerged as a promising tool in clinical hematology to ensure timely diagnosis, precise risk stratification, and treatment success. In this review, we summarize the current advances, limitations, and challenges of AI models and raise potential strategies for improving their performance in each sector of the MICM pipeline. Finally, we share perspectives, highlight future directions, and call for extensive interdisciplinary cooperation to perfect AI with wise human-level strategies and promote its integration into the clinical workflow.

OBJECTIVE: To compare the protective effects of electroacupuncture (EA) and transcutaneous electrical acupoint stimulation (TEAS) during pregnancy on maternal immune activation (MIA)-induced adverse pregnancy outcomes, fetal developmental defects, and neuropsychological behavior abnormalities in offspring mice. METHODS: Eighty pregnant C57BL/6 mice were randomly divided into 5 groups: control, model, EA, TEAS, and sham-stimulation groups, 16 mice in each group. MIA models were replicated on the day 12.5 of pregnancy via tail intravenous injection with polyinosinic-polycytidylic acid. On the second day of modeling success, in the EA and TEAS groups, the interventions were delivered at bilateral "Zusanli" (ST36), with a frequency of 2 Hz, a current of 0.5 mA, and for 20 min each day in the pregnant mice; and the interventions lasted 6 days. Body mass and fertility indexes of pregnant mice, and the development indexes of offspring mice were recorded. Liquid phase suspension chip technology was used to detect the levels of cytokines and chemotactic factors in the serum of pregnant mice and and fetal brain of offspring mice. Flow cytometry was adopted to detect the proportion of the subgroups and subtypes of spleen T lymphocytes and macrophages in pregnant mice. Using the open field test, prepulse inhibition (PPI) test and Morris water maze, the spatial learning and memory were assessed in offspring mice. Immunofluorescence staining was used to detect microglial count in the medial prefrontal cortex (mPFC) in offspring mice. RESULTS: Compared with the control group, the model group showed a reduced body mass of pregnancy mice (P<0.01), smaller litter size and fewer live births (P<0.01, P<0.05), the increase in dead birth and the decrease in offspring survival rate (P<0.05, P<0.01). When compared with model group, in the EA group and the TEAS group, the body mass of pregnancy mice rose (P<0.05), litter size and live births increased (P<0.05, P<0.01), the dead birth was reduced and the offspring survival rate higher (P<0.05). In comparison with the control group, the model group showed the increase in the levels of monocyte chemotactic protein-1 (MCP-1), interleukin-6 (IL-6), γ-interferon (IFN-γ) in the serum of pregnant mice, and spleen M1 macrophage proportion (P<0.01, P<0.05), and the decrease in spleen M2 macrophages of pregnant mice (P<0.01); and the increase in MCP-1 and IL-6 in fetal brain of offspring mice (P<0.05). Compared with the model group, the EA group and the TEAS group showed the decrease in MCP-1, IL-6 and IFN-γ, and spleen M1 macrophage proportion (P<0.01, P<0.05), and the increase in spleen M2 macrophages of pregnant mice (P<0.01, P<0.05) ; and the decrease in MCP-1 and IL-6 in fetal brain of offspring mice (P<0.05). Compared with the control group, in the model group, the total movement distance, escape incubation were extended (P<0.05, P<0.01), the frequency of entering the central area and crossing the platform decreased, and the activity duration in central area was shortened (P<0.05, P<0.01), the average speed rose (P<0.05), PPI%, the percentage of target quadrant swimming time in the total time and that of target quadrant swimming distance in the total distance were reduced (P<0.05, P<0.01) in offspring mice. When compared with the model group, in the EA group and TEAS group, the total movement distance and escape incubation were shortened, the average speed was reduced (P<0.05), PPI% and the frequency of crossing the platform increased (P<0.05, P<0.01); the percentage of target quadrant swimming time in the total time and that of target quadrant swimming distance in the total distance rose (P<0.05, P<0.01) in the offspring mice. In the EA group, the frequency of entering the central area and the activity duration in central area were higher (P<0.05, P<0.01); and in the the TEAS group, the activity duration in central area were longer (P<0.05). When compared with the control group, in the model group, microglial count in mPFC was elevated in offspring mice (P<0.05). In comparison with the model group, the EA group and the TEAS group showed the decrease of microglial count in mPFC (P<0.05). CONCLUSION EA and TEAS at "Zusanli" (ST36) during pregnancy effectively improve in the pregnancy outcomes and fetal brain developmental abnormalities induced by infection, and attenuate neurodevelopmental defects and mental disorders of offspring mice through inhibiting inflammatory activation of microglia in mPFC.
Animals ; Female ; Pregnancy ; Electroacupuncture ; Acupuncture Points ; Mice ; Mice, Inbred C57BL ; Humans ; Male

Metabolic dysfunction-associated steatotic liver disease (MASLD) is the most prevalent cause of chronic liver disease worldwide, and its intricate pathogenesis presents challenges in the development of new drugs. As a common way of post-translational modification, acetylation regulates protein stability, enzyme activity, and subcellular localization, occurring extensively in MASLD-associated processes such as lipid metabolism, inflammatory response, and oxidative stress. In this paper, we comprehensively review the mechanism of acetylation in MASLD, analyze the expression levels of acetylases in liver tissues of MASLD patients from the gene expression omnibus (GEO), discuss the changes in relevant enzyme expression and mechanisms in animal models, and further explore the feasibility of targeting acetylation for MASLD treatment, in the hope of offering a new perspective for advancing drug discovery in the field of MASLD.