ObjectiveTo investigate the impact of melanoma-associated antigen D4 （MAGED4） on the malignant biological behavior of hepatocellular carcinoma （HCC）， providing experimental evidence for its evaluation as a potential therapeutic target in HCC. MethodsGenes positively correlated with MAGED4 expression in HCC were screened using the LinkedOmics database， followed by GO functional and KEGG pathway enrichment analyses. Endogenously low-expressing SMMC-7721 and Huh7 cells were selected， with normal hepatic LO2 cells serving as controls. Stable overexpression cell lines were established via lentiviral infection （MAGED4 overexpression group）， with RT-PCR and Western blot validating overexpression efficiency. The effects of MAGED4 overexpression on cell proliferation， cell cycle distribution， migration， and adhesion were systematically analyzed using CCK-8 assays， flow cytometry， scratch healing assays， Transwell migration assays， and cell adhesion assays. ResultsBioinformatics analysis indicated that MAGED4 co-expressed genes were significantly enriched in biological processes closely associated with tumor proliferation and metastasis， including cell-cell adhesion， extracellular matrix organization， and the PI3K-Akt signaling pathway. A stable MAGED4 overexpression cell model was successfully established. In vitro functional assays demonstrated that MAGED4-overexpressing SMMC-7721 and Huh7 cells exhibited a mesenchymal-like morphology， whereas LO2 cells maintained an epithelial-like morphology. Compared with the empty-virus control groups， the MAGED4-overexpressing groups showed significantly increased proliferative activity （48 h： t=3.887， 22.070， 8.866， all P<0.05； 72 h： t=3.231， 23.840， 9.848， all P<0.05； 96 h： t=5.123， 49.800， 5.909， P<0.05， P<0.01， P<0.05）， wound healing rates （12 h： F=40.90， 44.00， 200.50， all P<0.001； 24 h： F=112.00， 34.89， 249.20， all P<0.001）， and numbers of migrated cells （F=239.20， 59.79， 132.80， all P<0.001）， along with significantly decreased numbers of adherent cells （F=73.65， 85.91， 138.00， all P<0.001）， while no significant differences were observed in cell cycle phase distribution in all three cell lines. ConclusionMAGED4 enhances proliferation and migration while suppressing adhesion in both HCC and normal hepatocytes， however， induces mesenchymal-like morphological changes exclusively in HCC cells， suggesting that MAGED4 is capable of driving HCC progression and may serve as a potential therapeutic target for HCC.

Objective Compare the clinicopathological factors of microsatellite unstable(dMMR)and microsatellite stable(pMMR)in right colon cancer and analyze the factors affecting overall survival(OS)and disease-free survival(DFS)between the two groups.Methods Clinical data with right colon cancer of patients who underwent radical resection from January 1,2016 to December 31,2023 were retrospectively analyzed(a total of 247 patients were included,including 43 dMMR and 204 pMMR).Through propensity score matching,the baseline difference between the two groups was matched 1∶1,and a total of 86 cases were obtained,43 cases in dMMR and 43 cases in pMMR.The difference of relevant indicators between the two groups was analyzed,and univariate and multivariate Cox analysis was performed for OS and DFS.The survival curve was plotted,and the comparison of survival rates was conducted using the Log rank test.Results According to age,location,tumor length,shape,histological type,T,N,TNM stage,nerve invasion,and differentiation degree with 1∶1 matching(P＜0.05),there was no statistical difference in baseline data between dMMR and pMMR patients with right colon cancer(P＞0.05).It was found that 5-year OS of dMMR patients with right colon cancer was significantly better than pMMR(P＜0.05),and 5-year DFS dMMR was better than pMMR(P＜0.05).Protective factors affecting OS of dMMR right colon cancer included tumor length＜5 cm,T1-3,M0,Stage Ⅰ-Ⅲ,absence of intravascular cancer thrombi and absence of nerve invasion(P＜0.05).In survival analysis,M0 was compared with M,(95％vs.60％,P＜0.05).Stage Ⅰ-Ⅲ compared with stage Ⅳ(95％vs.54％,P＜0.05);No vascular cancer thrombus vs.cancer thrombus(94％vs.88％,P＜0.05).Protective factors affecting DFS of dMMR right colon cancer included age＜50 years,no mucous or sig-ring cells,M0,StageⅠ-Ⅲ,and no intravascular cancer thrombus(P＜0.05).In survival analysis,M0 was compared with M,(90％vs.63％,P＜0.05).Stage Ⅰ-Ⅲ compared with stage Ⅳ(92％vs.57％,P＜0.05);There was a significant difference between non-vascular cancer thrombus and vascular cancer thrombus(91％vs.77％,P＜0.05).Conclusion The patients with dMMR had higher OS and DFS than pMMR,which were not affected by clinical factors.

Ovarian follicle expansion is an important part of their growth and development into dominant follicles,and is regulated by a variety of molecules and signals,including follicular cavity formation,follicular fluid accumulation,and granulosa cell proliferation.Polycystic ovary syndrome(PCOS)is the most common reproductive endocrine disease in women,and patients mainly present with increased preantral follicles and polycystic ovarian lesions caused by inadequate ovarian follicle expansion.This review summarizes recent research developments concerning the physiological process of ovarian follicle expansion and the related regulatory factors and mechanisms.We also consider the possible factors restricting ovarian follicle expansion in patients with PCOS,to provide a theoretical basis for follicular dysplasia,ovulation disorders and other diseases caused by abnormal ovarian follicle expansion.

Traditional development of small protein scaffolds has relied on display technologies and mutation-based engineering, which limit sequence and functional diversity, thereby constraining their therapeutic and application potential. Protein design tools have significantly advanced the creation of novel protein sequences, structures, and functions. However, further improvements in design strategies are still needed to more efficiently optimize the functional performance of protein-based drugs and enhance their druggability. Here, we extended an evolution-based design protocol to create a novel minibinder, BindHer, against the human epidermal growth factor receptor 2 (HER2). It not only exhibits super stability and binding selectivity but also demonstrates remarkable properties in tissue specificity. Radiolabeling experiments with ^99mTc, ⁶⁸Ga, and ¹⁸F revealed that BindHer efficiently targets tumors in HER2-positive breast cancer mouse models, with minimal nonspecific liver absorption, outperforming scaffolds designed through traditional engineering. These findings highlight a new rational approach to automated protein design, offering significant potential for large-scale applications in therapeutic mini-protein development.

Objective To develop a rehabilitation exercise program for the patients with osteoporotic vertebral compression fractures.Methods An online search was performed across both Chinese and English databases and websites to retrieve literature on rehabilitation exercises and related therapeutic measures for patients with osteoporotic vertebral compression fractures.Two researchers independently screened the retrieved literature,evaluated the quality,and extracted relevant evidence.Semi-structured interviews were conducted among 15 patients at various rehabilitation phases about the requirements of rehabilitation.A preliminary exercise program was developed and verified for the validity and feasibility using Delphi method.Results A rehabilitation exercise program was formulated.The program included five periods of preoperative period,bedridden period after surgery,ambulation period(1 week after surgery),weeks 2-8 after surgery and weeks 9-12 after surgery covering six domains of exercise type,exercise items,exercise duration,exercise frequency,exercise intensity and exercise safety.The response rate and effective rate over the two rounds of expert consultation were both 100.00%,with an authority coefficient of 0.94.The Kendall's W coefficients for secondary indicators in the two rounds of expert consultation were 0.184 and 0.334,respectively(both P<0.001).Conclusion The exercise program developed in this study for the patients with osteoporotic vertebral compression fractures is scientifically reliable and reasonable.The two features of continuity and staged characteristics in rehabilitation process are fully considered,thereby it offers a guidance for clinical healthcare professionals as well as the patients in development of practical and effective rehabilitation exercise plans.

Objective:To elucidate the causal association between gut microbiota and osteoporosis, we conducted a two-sample Mendelian randomization(MR)analysis to evaluate the causality between gut microbiota and osteoporosis.Methods:We assessed genome-wide association study (GWAS)summary statistics for gut microbiota and osteoporosis in older patients to perform the MR analysis.Independent single nucleotide polymorphisms closely associated with 211 gut bacterial taxa were identified as instrumental variables.The summary statistic data for osteoporosis were obtained from the latest release from the FinnGen consortium as the outcome of interest.Two-sample MR was performed to evaluate the causal effect of gut microbiota on osteoporosis, including the inverse-variance-weighted(IVW)method, weighted median method, MR-Egger, mode-based estimation, and MR-PRESSO.A series of sensitivity analyses were conducted to validate the robustness of the results.The robustness of the estimation was assessed through a series of sensitivity analyses, including Cochran's Q test, MR-Egger intercept analysis, leave-one-out analysis, and funnel plot to evaluate the causal association. Results:The population primarily consisted of individuals from Europe, with a total sample size of 7 300, including 1 762 males and 5 538 females, and average ages of 65.6 years for males and 67.8 years for females, respectively.The mean ages of the 5 538 patients were 65.6 years for males and 67.8 years for females, respectively.Through MR analysis in this study, 10 intestinal flora(comprising 1 class, 7 genera, and 2 orders)were identified as being associated with senile osteoporosis.Notably, the phylum Firmicutes( OR=1.324, P=0.019), Clostridium difficile( OR=1.274, P=0.038), fecal coliforms( OR=1.565, P=0.009), Verrucomicrobium( OR=1.231, P=0.033), and Selenomonas( OR=1.324, P=0.019)may serve as risk factors for senile osteoporosis.Conversely, Coprococcus( OR=0.620, P=0.024), Haemophilus( OR=0.815, P=0.016), Prevotella( OR=0.813, P=0.027), Sellimonas( OR=0.858, P=0.034), and RF9( OR=0.821, P=0.021)may function as protective factors against senile osteoporosis. Conclusions:This study suggests a causal relationship between the Firmicutes phylum and its nine subgroups and senile osteoporosis, thereby providing new insights into the mechanisms by which gut microbiota mediate senile osteoporosis and confirming the significant role of gut microbiota in this condition.

Objective:To investigate the clinical effects of radiofrequency microneedling versus fire needle therapy on facial acne. Methods:Ninety patients with facial acne treated at Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine from January 2022 to December 2023 were included in this study. Each patient's facial areas were divided into a radiofrequency microneedling group and a fire needle group, with treatments conducted every 2 weeks for a total of two sessions. The improvement of skin lesions on both sides was assessed using the Global Acne Grading System (GAGS), while the Visual Analog Scale (VAS) was used to evaluate the severity of pain experienced by patients. The incidence of adverse reactions, recurrence rates, and patient satisfaction were statistically analyzed. Additionally, a reflective confocal microscope was used to observe the imaging characteristics of the lesions before and after treatment.Results:At 2 and 4 weeks post-treatment, GAGS results showed that the improvement rates for skin lesions in the radiofrequency microneedling group were 83.33% (75/90) and 90.00% (81/90), respectively, both of which were significantly higher than those in the fire needle group [70.00% (63/90), 78.89% (71/90), χ2 = 4.47, 4.22, P = 0.034, 0.040]. The incidence of pigmentation in the radiofrequency microneedling group was 4.44% (4/90), which was slightly higher than that in the fire needle group [2.22% (2/90)]. However, there was no significant difference in overall incidence of adverse reactions between the radiofrequency microneedling and fire needle groups [15.56% (14/90) vs. 20.00% (18/90), χ2 = 0.60, P = 0.436]. The recurrence rate was lower in the radiofrequency microneedling group [10.00% (9/90)] compared with the fire needle group [22.22% (20/90), χ2 = 4.97, P = 0.026]. The VAS score for the radiofrequency microneedling group was (4.12 ± 1.33) during the first treatment and (3.30 ± 1.05) during the second treatment, both of which were significantly higher than those in the fire needle group [(3.67 ± 1.12), (2.88 ± 0.86), t = 2.45, 2.93, P = 0.015, 0.004]. Patient satisfaction with treatment outcomes did not significantly differ between the radiofrequency microneedling and fire needle groups [92.22% (83/90) vs. 87.78% (79/90), χ2 = 0.98, P = 0.320]. Results from the reflective confocal microscope indicated that after treatment, the hair follicles in both the radiofrequency microneedling and fire needle groups exhibited a distinct funnel shape and oval appearance, along with a noticeable reduction in funnel diameter compared with pre-treatment observations. Additionally, there was a decrease in the central amorphous material, a reduction in the size of the bright edge zone, and a decline in the infiltration of inflammatory cells, with no significant differences observed between the two groups. Conclusions:For facial inflammatory acne lesions (such as papules and pustules), both radiofrequency microneedling and fire needle treatments have shown good effectiveness. The fire needle treatment leads to less pain, while radiofrequency microneedling has a lower recurrence rate, but it carries a slightly higher risk of hyperpigmentation.

Ovarian follicle expansion is an important part of their growth and development into dominant follicles,and is regulated by a variety of molecules and signals,including follicular cavity formation,follicular fluid accumulation,and granulosa cell proliferation.Polycystic ovary syndrome(PCOS)is the most common reproductive endocrine disease in women,and patients mainly present with increased preantral follicles and polycystic ovarian lesions caused by inadequate ovarian follicle expansion.This review summarizes recent research developments concerning the physiological process of ovarian follicle expansion and the related regulatory factors and mechanisms.We also consider the possible factors restricting ovarian follicle expansion in patients with PCOS,to provide a theoretical basis for follicular dysplasia,ovulation disorders and other diseases caused by abnormal ovarian follicle expansion.

Helicobacter pylori(H.pylori)can colonize the gastric mucosa of humans and animals,leading to persistent infection,chronic inflammation,and tissue damage.To understand the preva-lence of H.pylori in dogs,this study designed specific primers targeting a unique region of the H.pylori virulence gene vacA and established a SYBR Green Ⅰ-based real-time PCR(qPCR)method.The specificity,sensitivity,and repeatability of this method were evaluated.Using the es-tablished method,alongside conventional PCR and H.pylori stool antigen(HpSA)detection,we conducted an epidemiological survey of H.pylori infection in pet dogs in urban Hangzhou.The re-sults showed that the standard curve demonstrated a good linear relationship within the concentra-tion range of 3.26×106 to 3.26×101 copies/μL.The method specifically detected H.pylori,with a minimum detection limit of 3.26×101 copies/μL.The intra-and inter-assay coefficients of variation ranged from 0.84％to 1.87％and 0.96％to 1.93％,respectively.Applying this method to canine fe-cal samples collected in Hangzhou from 2023 to 2024 revealed an overall H.pylori positive rate of 25.21％.In comparison,the detection rates by conventional PCR and HpSA were 10.68％and 8.97％,respectively.These results indicate that the established qPCR method is suitable for detec-ting H.pylori infection in dogs,providing a rapid,sensitive,specific,and reproducible quantitative detection method.Additionally,this study confirms the significant prevalence of H.pylori infection in household dogs.Given the widespread dissemination of H.pylori and its potential zoonotic risk,So recommend implementing risk communication and ongoing regional monitoring programs to protect the health of both humans and pets.

Objective To explore the work experiences and needs of nutrition specialist nurses,so as to provide a basis in career development and management practices among the nutrition specialist nurses.Methods A method of descriptive phenomenological research was adopted in this study.Purposive sampling was used to select 12 nutrition specialist nurses who came from different regions across the country with over one year of clinical working experiences as well as had completed the specialist training and been qualified by the Nutrition Committee of Guangdong Nursing Association between January and February 2024.Semi-structured on-line interviews were conducted among the participants and the data acquired from the interviews were analysed and summarised.Themes were extracted using NVivo12 and Colaizzi's seven-step analysis from the data from interview.Results Three main themes and eight sub-themes were identified regarding the work experience of nutrition specialist nurses:growth and gains(continuously learning in nutrition therapy-related knowledge to enhance professional skills;actively practice in nutrition therapy to achieve the self-worth);sense of responsibility and mission(increased awareness of responsibility and proactive services;sense of mission to cultivate talents and promotion of the development of nutrition nursing discipline);challenges(lack of an established role in nutrition nursing,conflict in responsibilities;insufficient communication in a medical team regarding nutrition therapy;limited resource to support the development of nutrition nursing;and the professional knowledge and skills unable to fully meet the job demands).Conclusion Hospitals managers should strengthen the support and training for nutrition specialist nurses,clarify their roles,leverage their professional value and promote the career development.