ObjectiveThis paper aims to verify the therapeutic effect of Cranial Painkiller pills' extract powder prepared by the new process on the rat's trigeminal neuralgia model caused by infraorbital injection of Talci Pulvis， evaluate its potential clinical application value， and compare the therapeutic effect with that of Cranial Painkiller granules， so as to provide data support for the application of the Cranial Painkiller pills' extract powder and precise treatment. MethodsThe rat's trigeminal neuralgia model was constructed by infraorbital injection of Talci Pulvis， and the rats were randomly divided into the normal group， model group， carbamazepine group （60 mg·kg^-1）， Cranial Painkiller granules group （2.70 g·kg^-1）， and low， medium， and high dosage groups of Cranial Painkiller pills' extract powder （1.35， 2.70， 5.40 g·kg^-1） according to the basal mechanical pain thresholds， and there were 10 rats in each group. The drug was administered by gavage to each group 2 h after modeling， and distilled water was given by gavage to the normal and model groups under the same conditions once a day for 10 d. Von Frey brushes were used to measure mechanical pain thresholds in rats. Hematoxylin-eosin （HE） staining was used to detect pathological changes in the trigeminal ganglion， and enzyme-linked immunosorbent assay （ELISA） was used to detect the inflammatory factors interleukin-1 （IL-1）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， and tumor necrosis factor-α （TNF-α） levels in rat serum， as well as neuropeptide substance P （SP） and β-endorphin （β-EP） levels in rat brain tissue. Western blot technique was used to detect the levels of NLRP3， ASC， Caspase-1， and OTULIN proteins in rat brain tissue. ResultsCompared with the normal group， the pain threshold of rats in the model group showed a continuous significant decrease （P<0.01）. The pathological damage of brain tissue was significant （P<0.01）， and the inflammatory levels of IL-1， IL-6， IL-8， and TNF-α in serum were significantly elevated （P<0.01）. The level of the SP in the brain tissue was significantly elevated （P<0.01）， and the level of β-EP was significantly reduced （P<0.01）， while the level of OTULIN was significantly reduced， and NLRP3， ASC， and Caspase-1 protein levels were significantly elevated （P<0.01）. After administration of the drug， compared with the model group， the pain threshold of each dose group of the Cranial Painkiller pills' extract powder and the Cranial Painkiller granules group significantly increased （P<0.01）. The inflammatory levels of IL-1， IL-6， IL-8， and TNF-α and SP levels significantly decreased （P<0.01）， and the β-EP levels were significantly elevated （P<0.01）， while the levels of OTULIN protein were significantly elevated （P<0.05， P<0.01）， and the levels of NLRP3， ASC proteins were decreased （P<0.01）in high dose Cranial Painkiller pills' extract powder. Meanwhile， compared with those in the model group， the trigeminal ganglion lesions of rats in the Cranial Painkiller pills' extract powder and Cranial Painkiller granules groups showed different degrees of improvement （P<0.05， P<0.01）. ConclusionThe Cranial Painkiller pills' extract powder has significant therapeutic effects on the rat model of trigeminal neuralgia induced by infraorbital injection of Talci Pulvis， and its mechanism is related to the improvement of OTULIN-regulated neuroinflammation.

ObjectiveThis paper aims to verify the therapeutic effect of Cranial Painkiller pills' extract powder prepared by the new process on the rat's trigeminal neuralgia model caused by infraorbital injection of Talci Pulvis， evaluate its potential clinical application value， and compare the therapeutic effect with that of Cranial Painkiller granules， so as to provide data support for the application of the Cranial Painkiller pills' extract powder and precise treatment. MethodsThe rat's trigeminal neuralgia model was constructed by infraorbital injection of Talci Pulvis， and the rats were randomly divided into the normal group， model group， carbamazepine group （60 mg·kg^-1）， Cranial Painkiller granules group （2.70 g·kg^-1）， and low， medium， and high dosage groups of Cranial Painkiller pills' extract powder （1.35， 2.70， 5.40 g·kg^-1） according to the basal mechanical pain thresholds， and there were 10 rats in each group. The drug was administered by gavage to each group 2 h after modeling， and distilled water was given by gavage to the normal and model groups under the same conditions once a day for 10 d. Von Frey brushes were used to measure mechanical pain thresholds in rats. Hematoxylin-eosin （HE） staining was used to detect pathological changes in the trigeminal ganglion， and enzyme-linked immunosorbent assay （ELISA） was used to detect the inflammatory factors interleukin-1 （IL-1）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， and tumor necrosis factor-α （TNF-α） levels in rat serum， as well as neuropeptide substance P （SP） and β-endorphin （β-EP） levels in rat brain tissue. Western blot technique was used to detect the levels of NLRP3， ASC， Caspase-1， and OTULIN proteins in rat brain tissue. ResultsCompared with the normal group， the pain threshold of rats in the model group showed a continuous significant decrease （P<0.01）. The pathological damage of brain tissue was significant （P<0.01）， and the inflammatory levels of IL-1， IL-6， IL-8， and TNF-α in serum were significantly elevated （P<0.01）. The level of the SP in the brain tissue was significantly elevated （P<0.01）， and the level of β-EP was significantly reduced （P<0.01）， while the level of OTULIN was significantly reduced， and NLRP3， ASC， and Caspase-1 protein levels were significantly elevated （P<0.01）. After administration of the drug， compared with the model group， the pain threshold of each dose group of the Cranial Painkiller pills' extract powder and the Cranial Painkiller granules group significantly increased （P<0.01）. The inflammatory levels of IL-1， IL-6， IL-8， and TNF-α and SP levels significantly decreased （P<0.01）， and the β-EP levels were significantly elevated （P<0.01）， while the levels of OTULIN protein were significantly elevated （P<0.05， P<0.01）， and the levels of NLRP3， ASC proteins were decreased （P<0.01）in high dose Cranial Painkiller pills' extract powder. Meanwhile， compared with those in the model group， the trigeminal ganglion lesions of rats in the Cranial Painkiller pills' extract powder and Cranial Painkiller granules groups showed different degrees of improvement （P<0.05， P<0.01）. ConclusionThe Cranial Painkiller pills' extract powder has significant therapeutic effects on the rat model of trigeminal neuralgia induced by infraorbital injection of Talci Pulvis， and its mechanism is related to the improvement of OTULIN-regulated neuroinflammation.

Trophoblast cells serve as the foundation for placental development. We analyzed published multiomics sequencing data and found that trophoblast cells highly expressed RRS1 compared to primitive endoderm and epiblast. We used HTR-8/SVneo cells for further investigation, and Western blot and immunofluorescence staining confirmed that HTR-8/SVneo cells highly expressed RRS1. RRS1 was successfully knocked down in HTR-8/SVneo cells using siRNA. Using IncuCyte S3 live-cell analysis system based on continuous live-cell imaging and real-time data, we observed that proliferation, migration, and invasion abilities were all significantly decreased in RRS1-knockdown cells. RNA-seq revealed that knockdown of RRS1 affected the gene transcription, and upregulated pathways in extracellular matrix organization, DNA damage response, and intrinsic apoptotic signaling, downregulated pathways in embryo implantation, trophoblast cell migration, and wound healing. Differentially expressed genes were enriched in diseases related to placental development. Consistent with these findings, human chorionic villus samples collected from spontaneous abortion cases exhibited significantly reduced RRS1 expression compared to normal controls. Our results highlight the functional importance of RRS1 in human trophoblasts and suggest that its deficiency contributes to early pregnancy loss.
Humans ; Trophoblasts/physiology* ; Cell Movement/genetics* ; Cell Proliferation/genetics* ; Female ; Pregnancy ; Abortion, Spontaneous/metabolism* ; Cell Line ; Placentation/genetics*

Objective:To investigate the impact of the number of examined lymph nodes (ELN) on survival outcomes in gastric cancer patients with postoperative pathological stage pN3b.Methods:This retrospective cohort study included 279 pN3b gastric cancer patients who underwent D2 gastrectomy at Nanfang Hospital, Southern Medical University (September 2008 to April 2023), with 35 patients receiving combination chemotherapy and anti-PD-1 therapy (immunotherapy group) and 244 receiving adjuvant chemotherapy alone (nonimmunotherapy group). Additionally, 422 patients with pN3b from the SEER database (2005 to 2020) were collected as an external validation cohort to determine the optimal cutoff value for the number of lymph nodes examined in the nonimmunotherapy group. The primary endpoints were overall survival (OS) and recurrence-free survival (RFS) in the nonimmunotherapy group of the Nanfang Hospital cohort, stratified by whether the number of examined lymph nodes was above or below the ELN optimal cutoff value. These findings were subsequently validated in the SEER cohort.Results:The optimal ELN cutoff value (34 nodes) was determined using X-tile software and by constructing an ELN-HR fitting model with inflection point identification. In the nonimmunotherapy group, patients with ELN >34 exhibited significantly prolonged survival compared to ELN ≤34 (median OS: 25.0 (95%CI:20.5-29.5) to 17.0 (95%CI:12.7-21.3) months, P=0.004; median RFS: 19.0 (95%CI:15.6-22.4) to 13.0 (95%CI:9.5-16.5) months, P=0.048). Multivariate Cox analysis also showed ELN >34 to be an independent protective factor for both OS (HR=0.576, 95%CI: 0.397-0.836) and RFS (HR=0.701, 95%CI: 0.492-0.998). In the SEER cohort, ELN >34 was associated with a 5-month OS extension (19 to 14 months, P=0.065), with multivariate analysis supporting its independent prognostic significance (HR=0.729, 95%CI: 0.580-0.915, P=0.006). Notably, in the immunotherapy group, patients with ELN >34 ( n=30) achieved a median OS of 41 months, but the median OS had not been reached in the ELN ≤34 group ( n=5) (1 death at 48 months). Conclusion:Higher ELN (>34) correlates with improved survival in nonimmunotherapy-treated pN3b gastric cancer patients. However, in pN3b gastric cancer patients treated with immunotherapy, the optimal ELN threshold requires further exploration to determine.

AIM:This study aims to investigate the role of histone methyltransferase SET and MYND domain containing 2(SMYD2)in facilitating renal fibrosis through the macrophage-myofibroblast transition in diabetic kidney dis-ease(DKD).METHODS:(1)C57BL/6J mice were intraperitoneally administered 55 mg/kg of streptozotocin to induce diabetes mellitus(DM).The experimental groups were categorized as follows:normal control,DM(20 weeks),DM(28 weeks),and DM(36 weeks).Blood glucose(BG),serum creatinine(SCr)and blood urea nitrogen(BUN)levels were determined using a biochemical analyzer.Hematoxylin-eosin(HE)staining and Masson staining were performed to assess morphological and fibrotic changes in renal tissues.Western blot analysis was used to measure the protein levels of SMYD2,histone H3 lysine 4 trimethylation(H3K4me3),arginase-1,matrix metalloproteinase 9(MMP9),collagen type Ⅰ(Col Ⅰ)and α-smooth muscle actin(α-SMA).Immunofluorescence staining was conducted to examine the localization and expression of F4/80,α-SMA,SMYD2,CD86,CD206 and CD163.(2)Mouse monocyte/macrophage RAW264.7 cells were cultured in vitro and assigned to groups as follows:normal glucose(NG)+negative control siRNA(siNC),high glucose(HG)+siNC,NG+SMYD2 siRNA(siSMYD2),and HG+siSMYD2.Western blot analysis was used to assess the expression of relevant proteins.RESULTS:(1)Compared with normal control group,the levels of BG,SCr and BUN were significantly elevated in DM(28 weeks)and DM(36 weeks)groups(P＜0.05).Renal tissue exhibited tubular atro-phy,dilation,and collagen fiber deposition.The levels of H3K4me3,arginase-1,MMP9,Col Ⅰ and α-SMA proteins were up-regulated(P＜0.05).The CD86,CD206,CD163 and F4/80 were primarily localized in the interstitial macrophages of the renal tubules,α-SMA was predominantly detected in the renal interstitium,and SMYD2 was mainly expressed in renal tubular epithelial cells and the renal interstitium.(2)Compared with NG+siNC group,the protein levels of SMYD2,H3K4me3,arginase-1,CD163,Col Ⅰ,α-SMA,transforming growth factor-β1(TGF-β1)and p-Smad3 in the cells of HG+siNC group were significantly increased(P＜0.05).Knockdown of SMYD2 resulted in a reduction of these indicators(P＜0.05).CONCLUSION:The SMYD2 protein appears to facilitate renal fibrosis in DKD by promoting the macrophage-myofibroblast transition,potentially through the modulation of TGF-β1/Smad3 signaling pathway.

Objective:To investigate the role of rheum tanguticum polysaccharide(RTP)in treating ulcerative colitis(UC)by improving intestinal flora.Methods:UC model was established,rats were divided into normal control group,model group and RTP treatment group.The integrity of colonic mucosa,submucosa and muscularis were observed by HE staining.Expressions of TNF-α,IL-6,IL-8 and IL-1β in serum were assessed by ELISA.16S rRNA sequencing was performed to analyze the composition and differences of intestinal flora in rats.Results:UC rats had symptoms such as loose stools,blood in the stool,anal filth,chills and mental malaise.RTP treatment could significantly improve the symptoms and reduce DAI scores(P<0.05).HE staining showed that the colonic struc-ture of UC rats was necrotic and inflammatory cells infiltrated submucosa.RTP treatment could significantly promote the repair and regeneration of mucosal epithelium.RTP could significantly inhibit inflammatory response and downregulate the expression of inflam-matory factors in UC rats(P<0.05).Composition and abundance of intestinal flora in UC rats were significantly different from that of normal control group.Staphylococcus was significantly increased,while Collinsella,Allobaculum,Chlamydia and Holdemanella were significantly decreased.RTP treatment could improve intestinal flora,reduce the abundance of Staphylococcus,and increase the abundance of Collinsella,Allobaculum,Chlamydia and Holdemanella.Conclusion:RTP can be used to treat UC in rats by inhibiting inflammation and improving intestinal flora.

Objective:To investigate the role of rheum tanguticum polysaccharide(RTP)in treating ulcerative colitis(UC)by improving intestinal flora.Methods:UC model was established,rats were divided into normal control group,model group and RTP treatment group.The integrity of colonic mucosa,submucosa and muscularis were observed by HE staining.Expressions of TNF-α,IL-6,IL-8 and IL-1β in serum were assessed by ELISA.16S rRNA sequencing was performed to analyze the composition and differences of intestinal flora in rats.Results:UC rats had symptoms such as loose stools,blood in the stool,anal filth,chills and mental malaise.RTP treatment could significantly improve the symptoms and reduce DAI scores(P<0.05).HE staining showed that the colonic struc-ture of UC rats was necrotic and inflammatory cells infiltrated submucosa.RTP treatment could significantly promote the repair and regeneration of mucosal epithelium.RTP could significantly inhibit inflammatory response and downregulate the expression of inflam-matory factors in UC rats(P<0.05).Composition and abundance of intestinal flora in UC rats were significantly different from that of normal control group.Staphylococcus was significantly increased,while Collinsella,Allobaculum,Chlamydia and Holdemanella were significantly decreased.RTP treatment could improve intestinal flora,reduce the abundance of Staphylococcus,and increase the abundance of Collinsella,Allobaculum,Chlamydia and Holdemanella.Conclusion:RTP can be used to treat UC in rats by inhibiting inflammation and improving intestinal flora.

Objective To investigate the diagnostic and therapeutic value of single-use mother-baby choledochoscope-assisted endoscopic retrograde appendicitis therapy in the treatment of acute uncomplicated appendicitis.Methods A retrospective analysis was conducted on the clinical data of 39 patients with acute uncomplicated appendicitis who underwent single-use mother-baby choledochoscope-assisted endoscopic retrograde appendicitis treatment at the Endoscopy center of the hospital from September 2022 to September 2024.Observe the endoscopic manifestations,the rate of maternal and child basket stone removal,the rate of appendiceal stent implantation,the technical success rate,the clinical success rate,the operation time,the hospital stay,the incidence of complications,the visual analogue scale(VAS)score 6 hours after the operation,and the inflammatory indicators 24 hours after the operation.Results In 28 cases(71.8%),congestion and edema could be seen at the opening of the appendix under colonoscopy.In 10 cases(25.6%),pus could be seen flowing out of the opening of the appendix under colonoscopy.In 32 cases(82.1%),a large amount of pus could be seen in the lumen of the appendix under subscopy.In 20 cases(51.3%),appendiceal fecalith could be seen in the lumen of the appendix under subscopy.The technical success rate of single-use mother-baby choledochoscope-assisted endoscopic retrograde appendicitis treatment was 100.0%(39/39).The operation time was(21.08±7.49)min;Hospital stay:(3.97±2.08)days;Eight cases(20.5%)of patients underwent endoscopic maternal basket stone removal.Appendiceal stent implantation was performed in 14 cases(35.9%)of patients.The clinical success rate is 97.4%(38/39).One patient's clinical symptoms and inflammatory indicators did not improve after the operation,and was transferred to the surgery department for appendectomy.The VAS score of 38 patients was less than 3 points 6 hours after the operation,and the abdominal pain symptoms were significantly relieved.The white blood cell count and the percentage of neutrophils 24 hours after the operation decreased significantly compared with those before the operation,and the differences were statistically significant(P<0.05).None of the 39 patients had complications.The postoperative follow-up was(5.94±4.03)months,and recurrence occurred in 3 cases(7.7%).Conclusion single-use mother-baby choledocoscope-assisted endoscopic retrograde appendicitis therapy is safe and effective in the diagnosis and treatment of acute uncomplicated appendicitis,which is worthy of further promotion and popularization in clinical practice.

Objective:To investigate the clinicopathological features, diagnosis, genetic alterations, and biological behaviors of hamartomatous inverted hyperplastic polyp (HIHP) in the gastrointestinal tract.Methods:The clinical, sonographic, endoscopic and pathologic data of 10 HIHP cases diagnosed at the First Affiliated Hospital of Air Force Medical University, Xi′an, China from January 2013 to March 2024 were collected. Their clinicopathological features and histological morphology were analyzed. The cases were further divided into 3 histologic subtypes. Follow-up information was collected to analyze the relationship between histological subtype and prognosis.Results:There were 5 males and 5 females in this cohort. The age of onset was 45-68 years, with a median age of 60.5 years. The polyp-involved sites included 2 cases in gastric fundus, 6 cases in gastric body, 1 case in gastric antrum, and 1 case in duodenum. Digestive endoscopy showed mucosal protrusion lesions in all cases, except 1 case (case 10) of shallow depression on the surface, with the maximum diameter ranging 0.5-2.5 cm. Endoscopic ultrasonography showed multilocular cystic low-density shadows, with septal enhancement (case 4). The preoperative clinical diagnosis was gastric polyp, ectopic pancreas or gastrointestinal stromal tumor. Two cases showed type 1 morphology (i.e., connected with the mucosa, with clear smooth muscle boundaries). One of them (case 10) had a clear opening to form a vase-like morphology, while the other (case 4) had no obvious opening with the surface mucosa. Three cases showed type 2 morphology (i.e., not connected with the mucosa, with clear smooth muscle boundaries). Five cases showed type 3 morphology (i.e., not connected with the mucosa, without clear smooth muscle boundaries or hyperplastic smooth muscle that separated hyperplastic glands showing lobular configuration). Among them, one case of duodenal lesions (case 9) showed gastric type gland hyperplasia and expansion, including gastric fossa, gastric fundic gland and pyloric gland, with various arrangement and combination, accompanied by smooth muscle hyperplasia. In case 10, there was leiomyomatous proliferation in the stroma. The cases 2 and 4 had atypical glandular structures and cell morphology, but immunohistochemistry showed wild-type expression pattern of p53 and a Ki-67 proliferation index of less than 1%, suggesting that it was reactive atypia secondary to inflammation. The results showed that 3 cases had different gene mutations, and no recurrent gene change was identified. All patients survived without disease during the follow-up period of 1-130 months.Conclusions:HIHP is a benign lesion and has no consistently detectable genetic alterations. The histological characteristics of gastrointestinal polyps are complex. Especially, the types 1 and 3 of HIHP have unique gross and microscopic features, which require combination of proper endoscopic sampling and histological examination to correctly classify them.

Objective:To observe any impact of supplementing conventional rehabilitation training with electrothermal acupuncture on the upper limb motor functioning of hemiplegic stroke survivors.Methods:Thirty-six hemiplegic persons who had recently survived a stroke were divided at random into an electrothermal acupuncture group ( n=18) and a control group ( n=18). Both groups received conventional rehabilitation training, while the electrothermal acupuncture group was additionally provided with daily 30-minute sessions of electrothermal acupuncture, 6 days per week for 3 weeks. Before and after the treatment everyone′s upper limb motor functioning was quantified using the Fugl-Meyer assessment (FMA-UE), and their ability in the activities of daily living (ADL) was assessed using the modified Barthel index (MBI). The Hong Kong version of the functional test for the hemiplegic upper extremity (FTHUE-HK) was also administered. Surface electromyography (sEMG) was employed to compute the co-contraction ratios (CRs) of the agonist to antagonist muscles during elbow flexion and extension. Results:After the treatment, both groups exhibited significant improvements in their average FMA-UE, FTHUE-HK and MBI results, but with significantly greater improvement in the electrothermal acupuncture group. The root mean square values of the wrist flexor and extensor and elbow flexor muscles during flexion showed significant improvement in both groups, with the average values in the electrothermal acupuncture group significantly better than those of the control group. The CR values during elbow extension in the electrothermal acupuncture group demonstrated significant improvement compared to before the treatment.Conclusions:Supplementing conventional rehabilitation measures with electrothermal acupuncture can better increase the strength of the elbow and wrist extensor muscles in the early days after a stroke. It promotes the recruitment of muscle fibers in the extensor muscle group, reduces abnormal coordination patterns in the elbow joint, and improves upper limb motor function, the practicality of hand function, and ability in the activities of daily living.