ObjectiveUlcerative colitis is a prevalent immunoinflammatory disease. Th17/Treg cell imbalance and gut microbiota dysregulation are key factors in ulcerative colitis pathogenesis. The actin cytoskeleton contributes to regulating the proliferation, differentiation, and migration of Th17 and Treg cells. Wdr63, a gene containing the WD repeat domain, participates in the structure and functional modulation of actin cytoskeleton. Recent research indicates that WDR63 may serve as a regulator of cell migration and metastasis via actin polymerization inhibition. This article aims to explore the effect of Wdr63 deletion on Th17/Treg cells and ulcerative colitis. MethodsWe constructed Wdr63^-/- mice, induced colitis in mice using dextran sulfate sodium salt, collected colon tissue for histopathological staining, collected mesenteric lymph nodes for flow cytometry analysis, and collected healthy mouse feces for microbial diversity detection. ResultsCompared with wild-type colitis mice, Wdr63^-/- colitis mice had a more pronounced shortening of colonic tissue, higher scores on disease activity index and histological damage index, Treg cells decreased and Th17 cells increased in colonic tissue and mesenteric lymph nodes, a lower level of anti-inflammatory cytokine IL-10, and a higher level of pro-inflammatory cytokine IL-17A. In addition, WDR63 has shown positive effects on maintaining intestinal microbiota homeostasis. It maintains the balance of Bacteroidota and Firmicutes, promoting the formation of beneficial intestinal bacteria linked to immune inflammation. ConclusionWdr63 deletion aggravates ulcerative colitis in mice, WDR63 inhibits colonic inflammation likely by regulating Th17/Treg balance and maintains intestinal microbiota homeostasis.

ObjectiveTo investigate the effects of Pinelliae Rhizoma Praeparatum （PRP） on lung tissue， gut microbiota， and short-chain fatty acid （SCFA） metabolism in a model of mice with cold fluid retention in the lung and explore its mechanism of action in resolving dampness and phlegm based on the interconnection between the lung and large intestine. MethodsFifty female ICR mice were randomly divided into a normal group， model group， positive control group （Xiaoqinglong granules， 6.5 g·kg^-1）， and high-dose and low-dose PRP decoction groups （3.0， 1.5 g·kg^-1）， with 10 mice in each group. A model of mice with cold fluid retention in the lung was established using ovalbumin （OVA） sensitization combined with cold-water immersion. Drug interventions were conducted from day 18 to day 33 for 15 consecutive days. The airway resistance value of the mice was measured using a non-invasive pulmonary function analyzer. Phlegm-resolving effects were evaluated via a microplate reader. Eosinophil and neutrophil counts in bronchoalveolar lavage fluid （BALF） were analyzed using an automated hematology analyzer. Serum levels of total immunoglobulin E （IgE）， interferon-γ （IFN-γ）， interleukin-4 （IL-4）， and BALF levels of interleukin-6 （IL-6） and interleukin-8 （IL-8） were quantified by enzyme-linked immunosorbent assay （ELISA）. Lung histopathology was assessed using hematoxylin-eosin （HE） staining. Immunohistochemistry （IHC） was employed to detect mucin 5AC （MUC5AC） and aquaporin 5 （AQP5） protein expression in lung tissue. Gut microbiota composition was analyzed via agarose gel electrophoresis， and fecal SCFA levels were measured by gas chromatography-mass spectrometry （GC-MS）. ResultsCompared with the normal group， the model group exhibited significantly increased airway resistance value （RI） （P<0.05）， elevated eosinophil and neutrophil counts and IL-6 and IL-8 levels in BALF （P<0.05）， increased serum IgE and IL-4 levels （P<0.05）， with reduced IFN-γ levels （P<0.05）. It also showed thickened bronchial walls， widened alveolar septa， narrowed lumens， and mucus plugs in lung tissue， upregulated MUC5AC protein expression and downregulated AQP5 protein expression （P<0.05）， decreased relative abundance of beneficial gut microbiota （Firmicutes， Clostridia， Clostridiales， Lactobacillaceae， and Lactobacillus）， and increased abundance of harmful microbiota （Bacteroidetes， Bacteroidia， Bacteroidales， Muribaculaceae， and Muribaculum）. In addition， the model group presented reduced fecal SCFA levels （acetate， propionate， and butyrate） （P<0.05）. After the intervention of PRP decoction， compared to the model group， all drug administration groups showed decreased RI （P<0.05）， increased phenol red excretion， declined eosinophil and neutrophil counts and IL-6， IL-8， IgE， and IL-4 levels （P<0.05）， and improved IFN-γ levels （P<0.05） and lung pathology improved. The MUC5AC protein expression decreased （P<0.05）， and the AQP5 protein expression increased （P<0.05）. The disorder of gut microbiota was improved， and the diversity of gut microbiota was restored， with a significantly increased relative abundance ratio of beneficial microbiota （P<0.05） and a significantly reduced relative abundance ratio of harmful microbiota （P<0.05）. The SCFA levels （acetate， propionate， and butyrate） increased （P<0.05）. The efficacy indicators of serum inflammatory factors （IgE， IL-4， and IFN-γ）， phlegm-resolving effect， airway resistance， total pathological score， and the protein expression of MUC5AC and AQP5 were correlated with gut microbiota and SCFAs. ConclusionPRP decoction alleviates cold-phlegm syndrome by modulating the gut-lung axis， promoting beneficial gut microbiota， enhancing SCFA production， restoring the balance of gut microbiota， and suppressing respiratory inflammation. This study provides novel insights into the TCM theory of interconnection between the lung and large intestine.

ObjectiveTo investigate the effects of Pinelliae Rhizoma Praeparatum （PRP） on lung tissue， gut microbiota， and short-chain fatty acid （SCFA） metabolism in a model of mice with cold fluid retention in the lung and explore its mechanism of action in resolving dampness and phlegm based on the interconnection between the lung and large intestine. MethodsFifty female ICR mice were randomly divided into a normal group， model group， positive control group （Xiaoqinglong granules， 6.5 g·kg^-1）， and high-dose and low-dose PRP decoction groups （3.0， 1.5 g·kg^-1）， with 10 mice in each group. A model of mice with cold fluid retention in the lung was established using ovalbumin （OVA） sensitization combined with cold-water immersion. Drug interventions were conducted from day 18 to day 33 for 15 consecutive days. The airway resistance value of the mice was measured using a non-invasive pulmonary function analyzer. Phlegm-resolving effects were evaluated via a microplate reader. Eosinophil and neutrophil counts in bronchoalveolar lavage fluid （BALF） were analyzed using an automated hematology analyzer. Serum levels of total immunoglobulin E （IgE）， interferon-γ （IFN-γ）， interleukin-4 （IL-4）， and BALF levels of interleukin-6 （IL-6） and interleukin-8 （IL-8） were quantified by enzyme-linked immunosorbent assay （ELISA）. Lung histopathology was assessed using hematoxylin-eosin （HE） staining. Immunohistochemistry （IHC） was employed to detect mucin 5AC （MUC5AC） and aquaporin 5 （AQP5） protein expression in lung tissue. Gut microbiota composition was analyzed via agarose gel electrophoresis， and fecal SCFA levels were measured by gas chromatography-mass spectrometry （GC-MS）. ResultsCompared with the normal group， the model group exhibited significantly increased airway resistance value （RI） （P<0.05）， elevated eosinophil and neutrophil counts and IL-6 and IL-8 levels in BALF （P<0.05）， increased serum IgE and IL-4 levels （P<0.05）， with reduced IFN-γ levels （P<0.05）. It also showed thickened bronchial walls， widened alveolar septa， narrowed lumens， and mucus plugs in lung tissue， upregulated MUC5AC protein expression and downregulated AQP5 protein expression （P<0.05）， decreased relative abundance of beneficial gut microbiota （Firmicutes， Clostridia， Clostridiales， Lactobacillaceae， and Lactobacillus）， and increased abundance of harmful microbiota （Bacteroidetes， Bacteroidia， Bacteroidales， Muribaculaceae， and Muribaculum）. In addition， the model group presented reduced fecal SCFA levels （acetate， propionate， and butyrate） （P<0.05）. After the intervention of PRP decoction， compared to the model group， all drug administration groups showed decreased RI （P<0.05）， increased phenol red excretion， declined eosinophil and neutrophil counts and IL-6， IL-8， IgE， and IL-4 levels （P<0.05）， and improved IFN-γ levels （P<0.05） and lung pathology improved. The MUC5AC protein expression decreased （P<0.05）， and the AQP5 protein expression increased （P<0.05）. The disorder of gut microbiota was improved， and the diversity of gut microbiota was restored， with a significantly increased relative abundance ratio of beneficial microbiota （P<0.05） and a significantly reduced relative abundance ratio of harmful microbiota （P<0.05）. The SCFA levels （acetate， propionate， and butyrate） increased （P<0.05）. The efficacy indicators of serum inflammatory factors （IgE， IL-4， and IFN-γ）， phlegm-resolving effect， airway resistance， total pathological score， and the protein expression of MUC5AC and AQP5 were correlated with gut microbiota and SCFAs. ConclusionPRP decoction alleviates cold-phlegm syndrome by modulating the gut-lung axis， promoting beneficial gut microbiota， enhancing SCFA production， restoring the balance of gut microbiota， and suppressing respiratory inflammation. This study provides novel insights into the TCM theory of interconnection between the lung and large intestine.

Objective: To investigate the efficacy，safety and feasibility of regional lymph node dissection in laparoscopic radical surgery for bladder cancer under the guidance of indocyanine green fluorescence imaging. Methods: A total of 30 patients with muscle invasive bladder cancer (T2/T3NxM0) who volunteered to enter the clinical trial were randomly divided into the indocyanine green imaging guided laparoscopic regional lymph node dissection group (n=15) and the standard pelvic lymph node dissection group (n=15).The number of positive lymph nodes，operation time，intraoperative bleeding volume，incidence of lymph leakage，and tumor recurrence and metastasis rate 2-year after surgery were collected. Results: The number of positive lymph nodes was (4.20±1.66) and (4.60±1.72) respectively in the indocyanine green and standard groups，with no statistically significant difference (P>0.05).There were no statistically significant difference in the tumor recurrence and metastasis rates 2-year after surgery between the two groups (P>0.05).However，the indocyanine green group had shorter operation time，less intraoperative bleeding volume，and lower incidence of lymphatic leakage than the standard group (P<0.05). Conclusion: Indocyanine green fluorescence imaging guided laparoscopic lymph node dissection has comparable clinical efficacy to standard lymph node dissection，but with fewer complications.

This case report presents a 16-year-old male patient with deficiency of adenosine deaminase 2(DADA2). The patient had a history of Raynaud′s phenomenon with digital ulcers since childhood. As the disease progressed, the patient developed retinal vasculitis, intracranial hemorrhage, skin necrosis, severe malnutrition, refractory hypertension, and gastrointestinal bleeding. Genetic testing revealed compound heterozygous mutations in the ADA2 gene (paternal c.1072G > A pathogenic mutation + maternal c.1065C > A variant of unknown clinical significance). ADA2 enzyme activity was significantly reduced (0.1 U/L). A multidisciplinary treatment team developed an individualized plan to address key issues, including nutritional support, blood pressure control, rehabilitation, pain management, and balancing anticoagulation/bleeding risks. After systematic intervention, the patient′s condition showed partial improvement. This case reveals clinical challenges such as anticoagulation decisions and nutritional support of DADA2. It also emphasizes the importance of early molecular diagnosis, tumor necrosis factor-α inhibitor targeted therapy, and multidisciplinary collaboration in management, underlining the core value of precision medicine in rare disease management.

In the field of organ donation and transplantation, the determination of death faces the dilemma of the separation between medical standards and social cognition, which may hinder medical practice and lead to controversial legal evaluations. Establishing brain death criteria through the "hard law" model is the path currently adopted by many countries. However, this approach has limitations, such as the attribution of the right to define death in the law, the conflict between the technological iteration of brain death criteria and legal stability, and the neglect of the social construction of the concept of death. Based on the theory of gradual social engineering and social systems theory, establishing brain death criteria through the "soft law" model has both scientific adaptability and governance flexibility, and is conducive to cultivating social consensus on brain death criteria. In terms of implementation, it is essential to focus on the high-level positioning of the "soft law" formulation entities for brain death criteria, the professionalization of the implementation entities, the publicization of the formulation process, and the institutionalization of risk communication strategies. At the same time, it is necessary to strengthen the informed consent of close relatives and design a dispute resolution mechanism for misjudgments of brain death to guard against potential improper risks.

OBJECTIVE: To observe the effect of "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique on the relative protein expression of the signaling pathway of nucleotide-binding oligomerization domain-containing protein 1 (NOD1)/ receptor-interacting protein 2 (RIP2)/nuclear factor kappa-B (NF-κB) and the expression of proinflammatory cytokines in the rats with primary dysmenorrhea (PD), so as to explore the underlying mechanism of this acupuncture technique for pain alleviation in PD. METHODS: Thirty female SD rats of SPF grade with normal estrous cycle were randomized into a blank group, a model group and an acupuncture group, 10 rats in each one. Using the intraperitoneal injection with estradiol benzoate combined with oxytocin, PD model was prepared in the model group and the acupuncture group. In the acupuncture group, during model preparation, the intervention with "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique was delivered simultaneously, 20 min each time, once daily for consecutive 10 days. On day 11, within 30 min after the intraperitoneal injection with oxytocin, the writhing reaction (latency, frequency and score) was recorded; the morphology of uterine tissue was observed with HE staining, the contents of prostaglandin E₂ (PGE₂), prostaglandin F_2α (PGF_2α), interleukin (IL)-1β, IL-18, cyclooxygenase-2 (COX-2), and tumor necrosis factor-α(TNF-α) in the serum were detected using ELISA method; the relative protein expression of NOD1, RIP2, NF-κB p65, phosphorylation-NF-κB p65 (p-NF-κB p65) was detected in the uterine tissue using Western blot method; and the mRNA expression of NOD1, RIP2 and NF-κB p65 was detected with the quantitative real-time PCR employed. RESULTS: Compared with the blank group, in the model group, the writhing latency was prolonged (P<0.01), the writhing frequency and score increased (P<0.01) in the rats; the endometrial epithelial cells showed massive degeneration and necrosis, with severe endometrial edema and widespread shedding, combined with neutrophil infiltration; the serum PGE₂ content was dropped (P<0.01), while those of PGF_2α, IL-1β, IL-18, COX-2, and TNF-α elevated (P<0.01); the protein expression of NOD1, RIP2, NF-κB p65 and p-NF-κB p65, and the mRNA expression of NOD1, RIP2 and NF-κB p65 in uterine tissue increased (P<0.01). In comparison with the model group, in the acupuncture group, the writhing latency was prolonged (P<0.01), the writhing frequency and score were reduced (P<0.01) in the rats; there was less degeneration and necrosis of endometrial epithelial cells, with mild endometrial edema and very little neutrophil infiltration; the serum PGE₂ content increased (P<0.01), while those of PGF_2α, IL-1β, IL-18, COX-2, and TNF-α decreased (P<0.01); the protein expression of NOD1, RIP2, NF-κB p65 and p-NF-κB p65 and the mRNA expression of NOD1, RIP2 and NF-κB p65 in uterine tissue were dropped (P<0.05, P<0.01). CONCLUSION "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique can alleviate the pain symptom of PD rats, and its action mechanism may be related to inhibiting the active expression of NOD1/RIP2/NF-κB signaling pathway in the uterine tissue, thereby reducing the inflammatory response.
Animals ; Female ; Rats, Sprague-Dawley ; Rats ; Signal Transduction ; Dysmenorrhea/metabolism* ; NF-kappa B/metabolism* ; Acupuncture Points ; Humans ; Acupuncture Analgesia ; Nod1 Signaling Adaptor Protein/metabolism* ; Receptor-Interacting Protein Serine-Threonine Kinase 2/metabolism* ; Acupuncture Therapy

OBJECTIVE: To compare the clinical efficacy of jingjin needling combined with rehabilitation training and conventional acupuncture combined with rehabilitation training for post-stroke spastic paralysis. METHODS: A total of 60 patients with post-stroke spastic paralysis were randomly divided into an observation group and a control group, 30 cases in each one. Both groups received conventional rehabilitation training. In the observation group, jingjin needling was applied at tendon blockage points of the shoulder, elbow, wrist, hip, knee, and ankle. In the control group, conventional acupuncture was applied at Jianyu (LI15), Quchi (LI11), Hegu (LI4), Biguan (ST31), Fengshi (GB31), Taichong (LR3), etc. on the affected side. Treatment was given once daily, 5 days a week for 4 weeks in both groups. The scores of clinical spasticity index (CSI), modified Ashworth scale, modified Barthel index (MBI), and Fugl-Meyer assessment scale (FMA) were evaluated before and after treatment, and the onset time was compared between the two groups. RESULTS: After treatment, the scores of CSI and modified Ashworth scale were decreased compared with those before treatment (P<0.001), while the scores of MBI and FMA were increased compared with those before treatment (P<0.001) in the two groups. After treatment, the scores of CSI and modified Ashworth scale in the observation group were lower than those in the control group (P<0.05), the MBI score in the observation group was higher than that in the control group (P<0.05). There was no statistically significant difference in FMA scores between the two groups (P>0.05). The onset time of the observation group was earlier than that in the control group (P<0.05). CONCLUSION Both jingjin needling combined with rehabilitation training and conventional acupuncture combined with rehabilitation training can effectively treat post-stroke spastic paralysis, jingjin needling combined with rehabilitation training exhibits better therapeutic effect and rapider onset.
Humans ; Acupuncture Therapy ; Male ; Female ; Middle Aged ; Aged ; Stroke/complications* ; Adult ; Acupuncture Points ; Muscle Spasticity/etiology* ; Treatment Outcome ; Stroke Rehabilitation ; Paralysis/therapy* ; Combined Modality Therapy

OBJECTIVE: To explore the possible mechanism by which the "Zhibian" (BL54) toward "Shuidao" (ST28) acupuncture improves ovarian function in mice with poor ovarian response (POR) by observing its effect on gut microbiota. METHODS: A total of 35 SPF-grade C57BL/6 female mice were screened for normal estrous cycles using vaginal smears, and 30 mice were selected. Ten mice were assigned to the blank group, while the remaining mice were used to establish the POR model by intragastric administration of tripterygium wilfordii suspension. The successfully modeled mice were randomly divided into a model group and an acupuncture group, with 10 mice in each group. After modeling, the acupuncture group received the "Zhibian" (BL54) toward "Shuidao" (ST28) acupuncture method once daily for 20 minutes per session. Ovulation induction began the day after the intervention, and samples were collected after ovulation induction. Vaginal cytology was used to observe estrous cycle changes, and the number of oocytes obtained, ovarian wet weight, and ovarian index were recorded. Serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), and anti-Müllerian hormone (AMH) were detected using ELISA. HE staining was used to observe ovarian histology. Gut microbiota was analyzed using 16S rRNA gene sequencing technology. Western blot was used to detect the relative protein expression levels of Occludin and zonula occludens-1 (ZO-1) in colonic tissue. Correlation analysis was conducted among serum hormone indexes, the number of oocytes obtained, ovarian index and gut microbiota. RESULTS: Compared with the blank group, the model group showed a higher estrous cycle disorder rate (P<0.01), increased serum FSH and LH levels, and a higher LH/FSH ratio (P<0.01), while the number of oocytes obtained, ovarian wet weight, ovarian index, and serum E2 and AMH levels were significantly reduced (P<0.01). Compared with the model group, the acupuncture group showed a lower estrous cycle disorder rate (P<0.01), decreased serum FSH and LH levels, and a lower LH/FSH ratio (P<0.01), along with an increased number of oocytes obtained, higher ovarian wet weight, ovarian index, and elevated serum AMH and E2 levels (P<0.01, P<0.05). The blank group had a large number of well-developed primordial follicles, with abundant and closely arranged follicles at various stages. In the model group, there was a significant increase in the number of atretic follicles, a reduction in the number of follicles at various stages, and loosely arranged ovarian tissue. Compared with the blank group, the model group showed a significant decrease in the number of normal follicles (P<0.01) and an increase in the number of atretic follicles (P<0.01). The acupuncture group showed a reduction in atretic follicles and an increase in the number of follicles at various stages compared with the model group, with a significant increase in normal follicles (P<0.01) and a decrease in atretic follicles (P<0.01). Compared with the blank group, the model group exhibited reduced gut microbiota diversity and richness, with significantly lower Chao1 and Shannon indices (P<0.01), and a greater clustering distance from the blank group. The model group also showed an increase in the relative abundance of Firmicutes_D, Verrucomicrobiota, Paramuribaculum, Dubosiella, and Muribaculum (P<0.01, P<0.05), while the relative abundance of Firmicutes_A and the relative protein expression of Occludin and ZO-1 in colonic tissue were decreased (P<0.01). Compared with the model group, the acupuncture group showed improved gut microbiota diversity and richness, with increased Chao1 and Shannon indices (P<0.05), and a clustering distance closer to the blank group. The acupuncture group exhibited reduced relative abundance of Firmicutes_D, Verrucomicrobiota, and Muribaculum (P<0.05, P<0.01), while the relative abundance of Firmicutes_A and the relative protein expression of Occludin and ZO-1 were significantly increased (P<0.01, P<0.05). Correlation analysis indicated a relationship between gut microbiota and serum hormone indicators, as well as the ovarian index. Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis showed that the metabolic pathways of the intersecting species were related to amino acid biosynthesis and nucleotide metabolism. CONCLUSION The "Zhibian" (BL54) toward "Shuidao" (ST28) acupuncture method improves ovarian function in POR mice, and its mechanism may be related to regulating gut microbiota structure and maintaining intestinal barrier homeostasis.
Animals ; Female ; Gastrointestinal Microbiome ; Mice ; Acupuncture Therapy ; Mice, Inbred C57BL ; Humans ; Ovary/physiopathology* ; Acupuncture Points ; Follicle Stimulating Hormone/metabolism* ; Luteinizing Hormone/metabolism* ; Estrous Cycle ; Anti-Mullerian Hormone/blood*