Objective To establish a stable cell line expressing luciferase using U87MG cells,and to establish a nude mouse glioblastoma in situ transplantation tumor model,in order to provide a suitable efficacy evaluation model for anti-glioblastoma drugs.Methods A stable and highly expressing luciferase U87MG cell line(U87MG-Luc)was prepared by infecting U87MG with lentivirus,and its stability was verified by repeated passages.U87MG-Luc single-cell suspension,which has been amplified and digested in vitro,was injected into the caudate nucleus of the right brain of nude mice using a brain stereotaxic device and a micro constant flow pump.Using small animal live imaging technology to observe the tumor formation and growth of U87MG-Luc inoculated,and confirm appropriate modeling conditions.Subsequently,the U87MG-Luc brain in situ tumor model was constructed to validate the efficacy of the first-line clinical treatment drug temozolomide,including indicators such as tumor growth status,animal weight,and survival time.Results Successfully established and screened U87MG-Luc stable cell line using lentiviral infection system.The luciferase expressed by stable strains was linearly related to cell seeding density,and the in vitro culture could stably express luciferase after continuous passage to the 21st generation.After inoculating U87MG-Luc cells at an appropriate concentration into the brain of nude mice,in situ brain tumor formation was achieved,with a high success rate(100％)of 2 × 105 cells/nude mouse.On the fifth day after inoculation,the brain tumor remained stable with no metastasis,and the tumor growth trend was good thereafter.In the U87MG-Luc brain in situ tumor model constructed in this study,temozolomide significantly inhibited tumor growth and prolonged the survival of tumor bearing nude mice(P＜0.001).Conclusion A stable and highly expressing luciferase U87MG-Luc cell line was successfully constructed,transduced,and screened based on the lentiviral expression system,and U87MG-Luc nude mouse brain in situ tumor model was constructed to provide a suitable pharmacological evaluation model for the discovery of anti-glioma drugs.

Fifty whitespotted bamboo sharks (Chiloscyllium playgiosum) of both sexes were used to establish a large capacity variable domain of the new antigen receptor (VNAR) library with a total capacity of over 10⁹ colony-forming units (CFU). It was applied to screen VNARs against human serum albumin (HSA) and human transcription factor EB (TFEB), respectively. Meanwhile, VNAR libraries specific to HSA and TFEB with capacities above 10⁸ CFU were obtained following conventional immunization. These two approaches were systematically studied in terms of VNAR yield and composition. By comparing the VNAR sequences obtained from naïve and antigen-immunized libraries, we found that the complementary-determining region 3 (CDR3) of the former differs in composition from that of the latter. It shares a higher degree of homology with the naïve library. Meanwhile, the binding efficiency assessed by ELISA is also different between the naïve and antigen-immunized libraries. The binding of VNARs from the TFEB-immunized library appeared to surpass that observed with the naïve libraries, whereas the performance of VNARs from the HSA-immunized library indicated that both the immunized and naïve libraries for HSA had positive binding responses in polyclonal and monoclonal ELISA. The results are useful to develop novel diagnostic and therapeutic products based on shark VNARs.

OBJECTIVES: To analyze the changes in disease burden and risk factors of atrial fibrillation/atrial flutter (AF/AFL) in China and globally from 1990 to 2021 to provide epidemiological evidence for developing effective preventive measures for AF/AFL in China. METHODS: Based on data from the Global Burden of Disease Study 2021, we analyzed sex- and age-specific AF/AFL burden metrics across 204 countries or territories. Age-standardized incidence rate (ASIR), prevalence, mortality rate (ASMR), and disability-adjusted life-years (DALYs) rate (ASDR) were calculated. Joinpoint regression was employed to compute average annual percentage changes (AAPC) in trends. The risk factors were systematically evaluated and further validated using Mendelian randomization analysis. RESULTS: From 1990 to 2021, China experienced an increase in ASIR of AF/AFL (from 42.63 to 44.93 per 100 000), contrasting with the global decline. While China's ASMR decreased (from 4.93 to 4.33 per 100 000), global ASMR rose slightly (from 4.24 to 4.36 per 100 000). China's ASDR remained stable at 93.29 per 100 000, whereas global ASDR increased marginally (100.81 to 101.40 per 100 000). Significant sex disparities were observed: males showed higher ASIR and ASDR, while females exhibited greater ASMR and ASDR. China had higher incidence and prevalence but lower mortality and DALY rates compared to global averages. In 2021, the key risk factors for AF/AFL included elevated systolic blood pressure, high BMI, smoking, alcohol use, high-sodium diet, and low temperature. Gender-specific patterns emerged: smoking was predominant in males, whereas high BMI disproportionately affected females. CONCLUSIONS AF/AFL incidence and prevalence keep increasing in China, and its large population base and demographic aging pose significant public health challenges. Maintaining healthy lifestyle habits, including smoking cessation, alcohol moderation, and a low-sodium diet can help to lower AF/AFL incidence, and regular screenings is crucial for its early detection and treatment.
Humans ; China/epidemiology* ; Risk Factors ; Atrial Flutter/epidemiology* ; Atrial Fibrillation/epidemiology* ; Mendelian Randomization Analysis ; Male ; Female ; Global Burden of Disease ; Incidence ; Prevalence ; Middle Aged ; Disability-Adjusted Life Years ; Aged

Gongfa is an essential approach to prevent and treat diseases in traditional Chinese medicine(TCM),often used to prevent a disease before it arises.Guided by TCM and modern scientific theories,the Tuina(Chinese therapeutic massage)Gongfa prescription theory implements the principle,method,prescription,and form in clinical pattern-identified treatment to prescribe the corresponding Tuina Gongfa prescription,i.e.,to prescribe a basic Tuina Gongfa prescription,specifically for a systemic disease,and modify Gongfa forms based on the basic prescription according to different patterns.The Gongfa prescription for pulmonary diseases designs corresponding Gongfa forms from six perspectives:lifting Yang,securing the exterior,opening the orifices,soothing the chest,harmonizing the stomach,and regulating Qi to prevent and treat diseases.The application of the pulmonary Gongfa prescription indicates the potential to apply the Tuina Gongfa prescription theory for the clinical prevention,treatment,and rehabilitation of disorders of other systems,thereby fully realizing the unique role of TCM Gongfa.

ObjectiveTo investigate the potential machanism of Xiaozhong Zhitong Mixture （消肿止痛合剂， XZM） in the treatment of diabetes foot ulcer （DFU）. MethodsFifty SD rats were randomly divided into blank group， model group， XZM group， inhibitor group， XZM plus inhibitor group （combination group）， with 10 rats in each group. Except for the blank group， rats were fed with high-sugar， high-fat， high-cholesterol diet， intraperitoneally injected with streptozotocin， and subjected to skin defect to establish DFU model. After successful modeling， the XZM group and the combination group were given 1 ml/（100 g·d）of XZM by gavage， while the blank group， model group， and inhibitor group were all given an equal volume of 0.9% sodium chloride injection by gavage. Thirty minutes later， the inhibitor group and the combination group were intraperitoneally injected with 5 mg/（kg·d） of Notch1 inhibitor DAPT. All groups were treated once a day. After 14 days of administration， the skin tissue from the dorsal foot of the blank group rats and wound tissue from the other groups were collected. The pathological changes of granulation tissue in the wound were detected using hematoxylin eosin （HE） staining. The microvascular density （MVD） in wounds was detected through immunohistochemical staining. Real time fluorescence quantitative polymerase chain reaction （RT-PCR） and western blotting were used to detect the mRNA and protein levels of Notch1 homolog （Notch1）， Delta-like ligand 4 （Dll4）， Delta-like ligand 4 （VEGF）， and angiopoietin 2 （Ang-2）， respectively. ResultsHistological results showed that the epidermal structure in the dorsal foot skin tissue of the rats in the blank group was intact. In the wound tissue of the model group， the epidermis exhibited excessive keratinization， vacuolar cytoplasm， and a large number of inflammatory cells infiltrating the tissue， while in the XZM group， a large amount of scab formation was observed in the epidermis， with no significant inflammatory cell infiltration and a noticeable increase in fibroblasts. In the combination group and the inhibitor group， partial epidermal scab formation was observed in the wound tissue with a small amount of inflammatory cell infiltration. Compared to those in the blank group， the MVD in the wound tissue increased in the model group， as well as the mRNA expression and protein levels of Notch1 and Dll4， while VEGFA and Ang-2 mRNA expression and protein levels significantly decreased （P＜0.05 or P＜0.01）. Compared to those in the model group， the MVD in the wound tissue of all medication groups significantly increased， and the mRNA and protein levels of Notch1 and Dll4 decreased， while VEGFA and Ang-2 mRNA expression and protein levels increased （P＜0.05 or P＜0.01）. Compared to the XZM group， the inhibitor group and the combination group showed decreased MVD in wound tissue， increased Notch1 and Dll4 mRNA and protein levels， and decreased expression of VEGFA and Ang-2 mRNA and proteins （P＜0.05 or P＜0.01）. ConclusionXZM can effectively promote wound healing in DFU rats， and its mechanism of action may be related to the inhibition of Dll4/Notch1 signaling pathway in the wound tissue， therey promoting angiogenesis.

Gongfa is an essential approach to prevent and treat diseases in traditional Chinese medicine(TCM),often used to prevent a disease before it arises.Guided by TCM and modern scientific theories,the Tuina(Chinese therapeutic massage)Gongfa prescription theory implements the principle,method,prescription,and form in clinical pattern-identified treatment to prescribe the corresponding Tuina Gongfa prescription,i.e.,to prescribe a basic Tuina Gongfa prescription,specifically for a systemic disease,and modify Gongfa forms based on the basic prescription according to different patterns.The Gongfa prescription for pulmonary diseases designs corresponding Gongfa forms from six perspectives:lifting Yang,securing the exterior,opening the orifices,soothing the chest,harmonizing the stomach,and regulating Qi to prevent and treat diseases.The application of the pulmonary Gongfa prescription indicates the potential to apply the Tuina Gongfa prescription theory for the clinical prevention,treatment,and rehabilitation of disorders of other systems,thereby fully realizing the unique role of TCM Gongfa.

Objective To establish a stable cell line expressing luciferase using U87MG cells,and to establish a nude mouse glioblastoma in situ transplantation tumor model,in order to provide a suitable efficacy evaluation model for anti-glioblastoma drugs.Methods A stable and highly expressing luciferase U87MG cell line(U87MG-Luc)was prepared by infecting U87MG with lentivirus,and its stability was verified by repeated passages.U87MG-Luc single-cell suspension,which has been amplified and digested in vitro,was injected into the caudate nucleus of the right brain of nude mice using a brain stereotaxic device and a micro constant flow pump.Using small animal live imaging technology to observe the tumor formation and growth of U87MG-Luc inoculated,and confirm appropriate modeling conditions.Subsequently,the U87MG-Luc brain in situ tumor model was constructed to validate the efficacy of the first-line clinical treatment drug temozolomide,including indicators such as tumor growth status,animal weight,and survival time.Results Successfully established and screened U87MG-Luc stable cell line using lentiviral infection system.The luciferase expressed by stable strains was linearly related to cell seeding density,and the in vitro culture could stably express luciferase after continuous passage to the 21st generation.After inoculating U87MG-Luc cells at an appropriate concentration into the brain of nude mice,in situ brain tumor formation was achieved,with a high success rate(100％)of 2 × 105 cells/nude mouse.On the fifth day after inoculation,the brain tumor remained stable with no metastasis,and the tumor growth trend was good thereafter.In the U87MG-Luc brain in situ tumor model constructed in this study,temozolomide significantly inhibited tumor growth and prolonged the survival of tumor bearing nude mice(P＜0.001).Conclusion A stable and highly expressing luciferase U87MG-Luc cell line was successfully constructed,transduced,and screened based on the lentiviral expression system,and U87MG-Luc nude mouse brain in situ tumor model was constructed to provide a suitable pharmacological evaluation model for the discovery of anti-glioma drugs.

Objective:To understand the cognition of the family caregivers of terminal cancer patients on digital health intervention, to clarify their actual needs, and to analyze the obstacles to their acceptance of digital health intervention, so as to develop a digital health intervention plan for the family caregivers of terminal cancer patients.Methods:From February 2024 to March 2024, the family caregivers of 16 patients with terminal cancer in Hunan Cancer Hospital Pain Hospice Ward were selected by objective sampling, who met the inclusion and exclusion criteria were interviewed in a semi-structured manner about the cognition and needs of digital health intervention, and the interview contents were sorted and analyzed using the Colaizzi7 step method.Results:A total of 16 family caregivers of terminal cancer patients, 4 males and 12 females, aged 26-55 years, were interviewed. Four themes were distilled from the interview results: family caregivers of terminally cancer patients agree on the importance of digital health interventions; lack of awareness of digital health interventions; expectations of digital health interventions; and possible confounders affecting digital health interventions.Conclusions:The family caregivers of terminal cancer patients had insufficient awareness of digital health intervention, but all showed affirmation of the development of digital health intervention services. It is recommended to actively improve the basic conditions of digital health services, strengthen publicity, raise the level of awareness of the family caregivers, and positively overcome the relevant interfering factors, so as to gradually promote the development of digital health services.

Objective:To observe the clinical efficacy of neck Gongfa exercise in intervening people at high risk for cervical spondylosis. Methods:A total of 212 participants from 8 companies at high risk for cervical spondylosis were divided into two groups using the random number table method,with 105 participants in the control group receiving health education and 107 participants in the trial group receiving an additional neck Gongfa exercise.After successive 3-month interventions,the two groups were compared in terms of cervical soft tissue tension and neck disability index(NDI)score.The incidence of cervical spondylosis was observed 3 months later. Results:During the process,10 cases dropped out in the trial group,and the control group had 9 dropout cases.After the intervention,the cervical soft tissue tension value and NDI score improved in both groups(P<0.05)and showed significant differences between the two groups(P<0.05).At the 3-month follow-up,the trial group had a lower incidence rate of cervical spondylosis than the control group(P<0.05). Conclusion:For people at high risk for cervical spondylosis,neck Gongfa exercise can effectively improve cervical soft tissue tension and motor dysfunction and lower the incidence of cervical spondylosis in the short run.