ObjectiveTo decipher the mechanism by which Zuoguiwan （ZGW） treat hyperthyroidism in rats with kidney-Yin deficiency based on the dopamine receptor D4 （DRD4）/nicotinamide adenine dinucleotide phosphate （NADPH） oxidase 4 （NOX4） signaling pathway. MethodsThe rat model of kidney-Yin deficiency was induced by unilateral intramuscular injection of dexamethasone （0.35 mg·kg^-1）. After successful modeling， the rats were randomized into model， methimazole （positive control， 5 mg·kg^-1）， low-， medium-， and high-dose （1.85， 3.70， 7.40 g·kg^-1， respectively） ZGW， and normal control groups. After 21 days of continuous gavage， the behavioral indexes and body weight changes of rats were evaluated. The pathological changes of the renal tissue were observed by hematoxylin-eosin staining. The serum levels of thyroid hormones ［triiodothyronine （T₃）， thyroxine （T₄）， thyroid-stimulating hormone （TSH）］， renal function indexes ［serum creatine （Scr） and blood urea nitrogen （BUN）］， energy metabolism markers ［cyclic adenosine monophosphate （cAMP） and cyclic guanosine monophosphate （cGMP）］， and oxidative stress-related factors ［superoxide dismutase （SOD）， malondialdehyde （MDA）， and NADPH）］ were measured by enzyme-linked immunosorbent assay （ELISA）. Western blot was employed to analyze the expression of DRD4， NOX4， mitochondrial respiratory chain complex proteins ［NADH：ubiquinone oxidoreductase subunit S4 （NDUFS4） and cytochrome C oxidase subunit 4 （COX4）］， and inflammation-related protein ［tumor necrosis factor-alpha （TNF-α）， interleukin-6 （IL-6）， p38 mitogen-activated protein kinase （MAPK）］ pathway in the renal tissue. ResultsCompared with the normal group， the model group showed mental malaise， body weight decreases （P<0.01）， inflammatory cell infiltration in the renal tissue， a few residual parotid glands in the thyroid， elevations in serum levels of T₃， T₄， Scr， BUN， cAMP， cAMP/cGMP， MDA， and NADPH （P<0.01）， down-regulation in protein levels of TSH， SOD， and DRD4 （P<0.05， P<0.01）， and up-regulation in expression of NOX4， p-p38 MAPK/p38 MAPK， and inflammatory factors （P<0.01）. Compared with the model group， ZGW increased the body weight （P<0.05， P<0.01）， reduced the infiltration of renal interstitial inflammatory cells， restored the thyroid structure and follicle size， lowered the serum levels of T₃， T₄， Scr， BUN， cAMP， cAMP/cGMP， MDA and NADPH （P<0.05， P<0.01）， up-regulated the expression of TSH， SOD and DRD4 （P<0.05， P<0.01）， and down-regulated the expression of NOX4， p-p38 MAPK/p38 MAPK， and inflammatory factors （P<0.05， P<0.01）. Moreover， high-dose ZGW outperformed methimazole （P<0.05）. ConclusionBy activating DRD4， ZGW can inhibit the expression of NOX4 mediated by the p38 MAPK pathway， reduce oxidative stress and inflammatory response， thereby ameliorating the pathological state of hyperthyroidism due to kidney-Yin deficiency. This study provides new molecular mechanism support for the clinical application of ZGW.

Objective To evaluate the safety and efficacy of a self-developed micro-normothermic machine perfusion (NMP) system (micro-perfusion device) for preserving isolated porcine limbs. Methods Five healthy Landrace pigs were selected, and their left and right forelimbs were randomly divided into the NMP group and static cold storage (SCS) group. The NMP group was perfused with the self-developed micro-perfusion device and polymerized hemoglobin perfusate for 32 hours at normothermia, while the SCS group was preserved at 4 ℃. Hemodynamic parameters such as perfusion pressure and flow were monitored. The pH value, partial pressure of oxygen (PO₂), lactic acid (Lac), creatine kinase (CK) and lactate dehydrogenase (LDH) in the perfusate were measured. Hematoxylin-eosin staining was used to assess the muscle tissue structure, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was employed to evaluate muscle cell apoptosis, and immunohistochemistry staining was applied to detect the expressions of tumor necrosis factor (TNF)-α and interleukin (IL)-6. A mixed-effects model was used to analyze the effects of time and treatment methods on tissue structure, cell apoptosis and inflammatory factors. Results The device could stably maintain a perfusion pressure of (69±15) mmHg and a flow rate of (117±42) mL/min. The pH value and electrolytes of the perfusate were generally stable, with PO₂ maintained at a high level. Lac was maintained at 5.38(3.81, 6.45) mmol/L, while CK and LDH increased over time. After 32 hours of perfusion in the NMP group, both the myocyte spacing and apoptosis rate were better than those in the SCS group. Mixed-effects model analysis showed that there were statistically significant differences in the effects of NMP treatment and SCS treatment on myocyte spacing and apoptosis rate per unit time (both P < 0.05). There were no statistically significant differences in TNF-α and IL-6 between the two groups, and mixed-effects model analysis showed no statistically significant differences in the effects of NMP treatment and SCS treatment on TNF-α and IL-6 per unit time (both P > 0.05). Conclusions The micro-perfusion device used in this study may achieve 32-hour normothermic preservation in a porcine limb amputation model, maintain basic metabolism and ionic homeostasis, reduce muscle structural damage and cell apoptosis without inducing additional inflammatory responses. This technology is expected to significantly extend the time window for replantation of amputated limbs in disaster rescue and long-distance transportation, providing an important technical basis for clinical translation and subsequent replantation research.

This paper summarizes the clinical experience in the syndrome differentiation and treatment of rosacea using the method of venting heat and resolving stagnation. It is considered that the key pathogenesis of rosacea is the accumulation of heat with stagnation. Accordingly， the method of venting heat and resolving stagnation is proposed， which vents and disperses constraint heat by applying approaches such as dredging defensive qi， clearing qi， venting ying （营） level， and cooling blood， while eliminating stagnation and masses through regulating qi， resolving dampness， dispelling phlegm and removing stasis. In clinical practice， a core prescription for venting heat and resolving stagnation is formulated， with flexible modifications made according to the clinical characteristics of different rosacea subtypes， including erythematotelangiectatic， papulopustular， phymatous， and ocular types， thereby providing therapeutic insights for the treatment of rosacea with traditional Chinese medicine.

Transfusion technicians constitute a core workforce in safeguarding blood safety, and the quality of their standardized training directly affects the operational efficiency of the blood services. At present, training programs for transfusion technicians across China have been developed in a diversified manner. Positive progress has been achieved in integrated theoretical and practical training, on-the-job faculty development, and regional collaborative training models. However, persistent challenges remain, including fragmented training providers, inconsistent training standards, limited coverage, and a disconnect between training and workforce utilization. Based on a review of current standardized training practice, this article systematically analyzes the structural challenges confronting current standardized training, and further proposes a framework for building a comprehensive training system, covering the definition and core components of standardized training, targeted improvement strategies and key assessment indicators. The study aims to facilitate the transformation of training for blood transfusion technicians in blood services from a project-based approach to an institutionalized model, and from fragmented implementation to a systematic framework.

Mammalian male germline development depends on a specialized gonadal somatic microenvironment. However, itsin vitro reconstruction remains challenging. Major barriers include the generation of appropriate supporting-cell lineages, organized tissue architecture, and culture conditions that sustain germ-cell maturation. Yoshino et al. established a mouse pluripotent stem cell-derived testicular organoid model. This model reconstitutes the sex-determining region Y (Sry)-SRY-box transcription factor 9 (Sox9)‑mediated sex-determination program and generates testicular somatic cell-like cells (TesLCs). When assembled with primordial germ cell-like cells (PGCLCs), TesLCs formed seminiferous tubule-like structures and supported male germline progression toward prospermatogonia and germline stem cell-like cells (GSCLCs). After transplantation into infertile recipient testes, organoid-derived GSCLCs produced fertilization-competent sperm and generated fertile offspring. The major significance of this work extends beyond sperm production. It integrates sex determination, germ-cell fate transition, and testicular niche self-organization into an experimentally tractable organoid system. This commentary discusses the technical strategy, sex-determination mechanism, value of testicular niche reconstruction, and translational limitations of this study. It also highlights its implications for in vitro gametogenesis, male infertility modeling, and reproductive medicine.

Objective To systematically evaluate the similarities between six-gene-modified pig red blood cells and human red blood cells in multiple dimensions, including cell morphology, hematological parameters, microbial safety and serological compatibility, and to verify the feasibility of using six-gene-modified pig red blood cells as an alternative for blood transfusion in cases of traumatic hemorrhage. Methods Blood samples from O-type human donor, six-gene-modified O-type pig and wild-type O-type pig were collected. The pig gene-modified phenotype was identified by flow cytometry. The morphology was observed using field emission scanning electron microscopy and the diameter was measured. Hematological tests, including blood routine, osmotic fragility test, adenosine triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) content and partial pressure of oxygen at 50% hemoglobin saturation (P50), were conducted to assess the deformability of red blood cells. Real-time fluorescent quantitative polymerase chain reaction was used to screen for zoonotic viruses, and cross-matching was performed using column gel method, condensation amine method and saline tube method. Results The six-gene-modified pig red blood cells successfully knocked out the α-Gal, Neu5Gc and Sda antigens and expressed hCD55. Wild-type pig red blood cells expressed these three antigens but did not express human complement regulatory proteins. All three types of red blood cells presented a biconcave discoid morphology. The diameter of O-type human red blood cells was larger than that of the six-gene-modified porcine red blood cells and wild-type pig red blood cells (all P < 0.05). The average red blood cell volume and average red blood cell hemoglobin of O-type humans were higher than those of the six-gene-modified pig red blood cells, and the distribution width-variable coefficient of O-type human red blood cells was lower than that of the six-gene-modified pig red blood cells (all P < 0.05). There were no statistically significant differences in the permeability fragility, ATP content, 2,3-DPG content, P50 and red blood cell deformability indicators of the three types of red blood cells (all P > 0.05). The screening for zoonotic viruses in human and animal cells of the six-gene-modified pigs and wild-type pigs was negative. The cross-matching results showed that in the column gel method and condensation amine method, the primary and secondary sides of the six-gene-modified pig red blood cells did not agglutinate with human red blood cells, while the wild-type pig red blood cells agglutinated with human red blood cells in all cross-matching methods. Conclusions The six-gene-modified pig red blood cells are highly similar to human red blood cells in terms of morphology, key hematological parameters, energy metabolism, oxygen-carrying capacity, mechanical properties, microbial safety and serological compatibility, providing an experimental basis for the clinical transformation of xenotransfusion.

This paper summarizes Professor WU Mianhua's clinical experience in the staged treatment of radiation-induced brain injury （RIBI） based on the stasis-heat theory. It is believed that radiation fire-toxin is the initiating pathogenic factor of RIBI， and the core pathogenesis consists of the intermingling of stasis and heat， obstruction of collaterals by wind-phlegm， dual deficiency of qi and yin， as well as impairment of essence and depletion of marrow. In the early stage， the disease is mainly manifested as stasis-heat with fluid retention and obstruction of collaterals by wind-phlegm， with the lesion located in the brain collaterals. The therapeutic principle is to cool the blood and dissipate stasis， extinguish wind and unblock collaterals， and simultaneously eliminate phlegm and promote urination， according to which self-designed Liangxue Tongluo Formula （凉血通络方） is selected for treatment.In the middle stage， the key pathological features are the binding of phlegm， stasis and heat， and consumption of kidney essence， with the lesion invading the kidney essence. The therapeutic principle is to resolve phlegm and remove stasis， clear and relieve latent heat， replenish essence and tonify marrow， and the self-designed Qinghua Tianjing Formula （清化填精方） is selected for treatment. In the advanced stage， the disease is mostly manifested as dual deficiency of qi and yin and toxin-induced marrow exhaustion， with the lesion involving the brain marrow. The therapeutic principle is to replenish qi and nourish yin， remove toxin and tonify marrow， and the self-designed Yangyin Jiedu Formula （养阴解毒方） is selected for treatment.

Objective To construct a nomogram for predicting the overall survival rate of patients with gallbladder cancer with positive regional lymph nodes (LNs). Methods Data of 1413 patients diagnosed with gallbladder cancer with regional LN metastases from 2000 to 2022 were extracted from the Surveillance, Epidemiology, and End Results (SEER) database of the National Cancer Institute in the United States. The patients were randomly divided into a training set and a validation set at a ratio of 7:3. The Cox proportional hazards regression model was used to explore the independent risk factors affecting the survival of patients and construct a nomogram. The receiver operating characteristic (ROC) curve, calibration curve, and clinical decision curve were drawn to evaluate the predictive ability of the model. Results Multivariate Cox analysis showed that age, T stage, M stage, grade, surgery, radiotherapy, and chemotherapy were independent risk factors affecting the overall survival rate of patients with gallbladder cancer with positive regional LNs (P<0.05). These factors were all included in the nomogram to construct the prediction model. The areas under the ROC curves for 6 months, 1 year, and 3 years in the training set were respectively 0.849, 0.825 and 0.821; those in the validation set were respectively 0.826, 0.828 and 0.795. The calibration curves showed good calibration in both the training set and the validation set. The clinical decision curve showed high clinical application value. Conclusion The constructed nomogram model has good predictive ability and can help clinicians predict the overall survival of patients with gallbladder cancer with positive regional LNs, facilitating individualized treatment.

Objective: To analyze the incidence and mortality trends of colorectal cancer in Tongxiang City, Zhejiang Province from 2015 to 2024, so as to provide the evidence for optimizing regional colorectal cancer prevention and control strategies. Methods: Data on colorectal cancer incidence and mortality in Tongxiang City from 2015 to 2024 were collected through the Zhejiang Provincial Chronic Disease Surveillance Information Management System. The crude incidence and crude mortality of colorectal cancer were calculated, and age-standardized rates were derived using the standard age structure from the Fifth National Population Census in 2000. The trends in incidence and mortality were analysed using the average annual percent change (AAPC). Results: From 2015 to 2024, the annual crude incidence of colorectal cancer in Tongxiang City was 47.89/105, showing an upward trend (AAPC=5.845%, P<0.05); the annual age-standardized incidence was 22.14/105, with no statistically significant trend (P>0.05). From 2015 to 2024, the annual crude incidence and annual age-standardized incidence of colorectal cancer among men in Tongxiang City were higher than those among women (both P<0.05). The crude incidence and age-standardized incidence for men showed upward trends (AAPC=4.920%, 2.799%, both P<0.05), while the trends in crude incidence and age-standardized incidence for women were not statistically significant (both P>0.05). The annual crude incidence of colorectal cancer in the ≥75 age group was relatively high at 173.37/105; The crude incidence in the 50-<65 age group showed an upward trend (AAPC=3.970%, P<0.05), while trends in crude incidence for other age groups were not statistically significant (all P>0.05). From 2015 to 2024, the annual crude mortality and annual age-standardized mortality of colorectal cancer in Tongxiang City were 18.74/105 and 7.58/105, respectively; trends in crude mortality and age-standardized mortality were not statistically significant (both P>0.05). The annual crude mortality and annual age-standardized mortality of colorectal cancer were higher in men than in women (both P<0.05); there were no statistically significant trends in crude mortality and annual age-standardized mortality for either men or women (all P>0.05). The annual crude mortality of colorectal cancer was higher in the ≥75 age group, at 131.73/105; there were no statistically significant trends in crude mortality across age groups (all P>0.05). Conclusions From 2015 to 2024, the incidence of colorectal cancer in Tongxiang City showed an upward trend, while the mortality remained relatively stable. Men and the middle-aged and elderly were the key target groups for the prevention and control of colorectal cancer.

Objective: To evaluate the feasibility of dynamic contrast-enhanced MRI (DCE-MRI) in differentiating tumor deposits (TDs) from metastatic lymph nodes (MLNs) in rectal cancer. Materials and Methods: A retrospective analysis was conducted on 70 patients with rectal cancer, including 168 lesions (70 TDs and 98 MLNs confirmed by histopathology), who underwent pretreatment MRI and subsequent surgery between March 2019 and December 2022. The morphological characteristics of TDs and MLNs, along with quantitative parameters derived from DCE-MRI (K trans , kep, and v e) and DWI (ADCmin, ADCmax, and ADCmean), were analyzed and compared between the two groups.Multivariable binary logistic regression and receiver operating characteristic (ROC) curve analyses were performed to assess the diagnostic performance of significant individual quantitative parameters and combined parameters in distinguishing TDs from MLNs. Results: All morphological features, including size, shape, border, and signal intensity, as well as all DCE-MRI parameters showed significant differences between TDs and MLNs (all P < 0.05). However, ADC values did not demonstrate significant differences (all P > 0.05). Among the single quantitative parameters, v e had the highest diagnostic accuracy, with an area under the ROC curve (AUC) of 0.772 for distinguishing TDs from MLNs. A multivariable logistic regression model incorporating short axis, border, v e, and ADC mean improved diagnostic performance, achieving an AUC of 0.833 (P = 0.027). Conclusion The combination of morphological features, DCE-MRI parameters, and ADC values can effectively aid in the preoperative differentiation of TDs from MLNs in rectal cancer.