Objective To investigate the effect of tritiated water on the immune system of zebrafish and its potential molecular mechanism. Methods Zebrafish embryos (2.5 to 3 hours post-fertilization [hpf]) were exposed to 3.7 × 10⁴ Bq/mL tritiated water (tritiated water group), and those exposed to E3 culture medium were used as the control group. The mortality rate, hatching rate, deformity rate, heart rate, body length, yolk sac area, neutrophil count in the tail, immune-related gene expression, and immune-related protein expression of zebrafish in the two groups were determined. Then transcriptome technology was used to further analyze the possible mechanism of tritiated water affecting the immune system of zebrafish. Results Compared with the control group, zebrafish at 72 hpf in the tritiated water group had no significant changes in the mortality rate, hatching rate, deformity rate, body length, and yolk sac area((t = 0.9045, 0.5000, 1.0000, 0.7238, 0.0337, P = 0.4169, 0.6433, 0.3739, 0.4785, 0.9735), but had significantly increased heart rate(t = 4.575，P = 0.002). At 4 days post-fertilization (dpf), the neutrophil count in the tail of zebrafish in the tritiated water group was significantly increased(t = 2.563，P = 0.0196), the mRNA expression of TNF-α was significantly decreased(t = 2.891, P = 0.045), the protein expression of nuclear factor-kappa B (NF-κB) was significantly increased(t = 3.848, P = 0.018), and the protein expression of NLRP3 was significantly decreased(t = 14.98, P = 0.001). At 7 dpf, the neutrophil count in the tail and the protein expression levels of NF-κB, NLRP3, and interleukin-1β were significantly decreased(t = 3.772, 7.048, 15.620, 4.423, P = 0.014, 0.002, 0.0001, 0.012). Transcriptome sequencing revealed that differentially expressed genes were mainly enriched in the “neutrophil activation” and “platelet activation pathways” at 4 dpf and in the “neutrophil apoptosis”, “ferroptosis”, and “necroptosis” pathways at 7 dpf. Conclusion Tritiated water exposure induces a temporally dynamic immune response in zebrafish, potentially affecting immune homeostasis by regulating neutrophil activation and apoptosis, as well as the expression of NF-κB and NLRP3.

OBJECTIVE: To observe the effects of antagonistic needling therapy on lower limb spasticity and the muscle morphology of the tibialis anterior and gastrocnemius in patients with stroke. METHODS: A total of 100 patients with post-stroke lower limb spasticity were randomly divided into an antagonistic needling group (50 cases, 1 case dropped out) and a routine acupuncture group (50 cases, 1 case dropped out). Both groups received basic treatment and rehabilitation training. The routine acupuncture group was treated with scalp acupuncture at anterior oblique line of vertex-temporal and vertex lateral line 1, combined with body acupuncture at Jianyu (LI15), Hegu (LI4), Zusanli (ST36), Taichong (LR3), etc. on the affected side, with Quchi (LI11) and Hegu (LI4), Zusanli (ST36) and Fenglong (ST40), Yanglingquan (GB34) and Taichong (LR3) connected to an electroacupuncture device, using disperse wave at 2 Hz of frequency. The antagonistic needling group used the same scalp and upper limb acupoints as the routine acupuncture group, with additional antagonistic needling on the lower limb at Yanglingquan (GB34), Qiuxu (GB40), Jiexi (ST41), and Xuanzhong (GB39) on the affected side, with Quchi (LI11) and Hegu (LI4), Yanglingquan (GB34) and Qiuxu (GB40), Jiexi (ST41), and Xuanzhong (GB39) connected to an electroacupuncture device, using disperse wave at 2 Hz of frequency. Both groups received treatment once daily for 6 consecutive days per course, with a total of 4 courses. The modified Ashworth scale (MAS), Holden functional ambulation classification (FAC), lower limb Fugl-Meyer assessment (FMA), composite spasticity scale (CSS), and musculoskeletal ultrasound parameters (thickness and fiber length of the tibialis anterior and gastrocnemius, and pennation angle of the gastrocnemius on both sides) were evaluated before and after treatment. Clinical efficacy was compared between the two groups. RESULTS: Compared before treatment, the MAS grades and CSS scores were decreased in both groups after treatment (P<0.01), with greater reductions in the antagonistic needling group (P<0.05, P<0.01). FAC grades and FMA scores were increased in both groups after treatment (P<0.01, P<0.05), with greater improvements in the antagonistic needling group (P<0.05). The muscle thickness, fiber length of the tibialis anterior, the muscle thickness, fiber length and pennation angle of the gastrocnemius on the affected side were improved in both groups after treatment (P<0.01), with greater improvements in the antagonistic needling group (P<0.01, P<0.05). On the unaffected side, these parameters were also increased after treatment in both groups (P<0.01, P<0.05), but the antagonistic needling group showed smaller increases than the routine acupuncture group (P<0.01, P<0.05). The total effective rate in the antagonistic needling group was 91.8% (45/49), higher than 81.6% (40/49) in the routine acupuncture group (P<0.05). CONCLUSION Antagonistic needling could effectively reduce spasticity, improve motor function, and enhance muscle structure in patients with post-stroke lower limb spasticity.
Humans ; Male ; Female ; Acupuncture Therapy ; Middle Aged ; Muscle Spasticity/pathology* ; Aged ; Stroke/physiopathology* ; Lower Extremity/physiopathology* ; Acupuncture Points ; Adult ; Muscle, Skeletal/pathology* ; Treatment Outcome

This paper introduces Professor SUN Shentian's clinical experience in the treatment of refractory facial paralysis with acupuncture and moxibustion. Professor SUN believes that the etiology of refractory facial paralysis is complex. Acupuncture and moxibustion treatment should be based on cortical localization, Baihui (GV20), lower 1/5 of motor area and brainstem area are selected, and repetitive transcranial acupuncture is applied. Under the ultrasonic positioning, acupuncture is performed on the starting and ending points of the mimetic muscles in different lesion sites. Combined with the TCM pathogenesis of refractory facial paralysis with deficiency of healthy qi and retention of pathogenic factors, acupuncture and moxibustion treatment takes strengthening the healthy qi and eliminating pathogenic factors as the core, and reuses the acupoints of yangming meridians (Yingxiang [LI20], Sibai [ST2], Dicang [ST4], Hegu [LI4], Zusanli [ST36], etc.) as the main acupoints to dredge the meridians. The main facial mimetic muscles and related collateral points are selected for cluster needling to dredge the collaterals. Acupuncture at Yangbai (GB14)-toward-Tongziliao (GB1), Sibai (ST2)-toward-Dicang (ST4), Dicang (ST4)-toward-Jiache (ST6) is applied and combined with the needle-sticking and lifting technique to nourishing tendons. Qihai (CV6) and Guanyuan (CV4) are selected for acupuncture before moxibustion. In addition, Professor SUN emphasizes that the three methods of kneading, acupuncture and moxibustion should be used in Yifeng (TE17), Qianzheng (Extra) and Xiaguan (ST7). Professor SUN combines TCM syndrome differentiation with modern technology, which has the advantages of accurate positioning and diverse techniques, and provides a new idea for the treatment of refractory facial paralysis.
Humans ; Moxibustion ; Acupuncture Therapy ; Facial Paralysis/therapy* ; Female ; Male ; Acupuncture Points ; Middle Aged ; Adult

OBJECTIVES: To investigate the protective effects and underlying mechanisms of Gynostemma pentaphyllum (GP)ethanol extract on motor dysfunction in a mouse model of Parkinson's disease (PD). METHODS: Eighty C57BL/6 male mice were randomly divided into five groups: control group, model group, levodopa group (positive control group), low-dose GP group, and high-dose GP group, with 16 mice per group. The PD model was induced by injection of 6-hydroxydopamine into the substantia nigra pars reticulata of the mice. Two weeks after 6-hydroxydopamine, positive control group received intraperitoneal injection of levodopa 10 mg·kg^－1·d^－1, while low-dose GP and high-dose GP groups received GP extract 100 or 200 mg·kg^－1·d^－1 orally for three weeks. After a 3-week-treatment, the effects of GP on motor dysfunction in 6-hydroxydopamine-induced PD were assessed using open field and CatWalk gait tests, while the effects on muscle strength were evaluated by forelimb grip strength. Immunofluorescence staining was used to detect the number of tyrosine hydroxylase (TH) positive neurons. The levels of dopamine and serotonin in the midbrain were determined by enzyme-linked immunosorbent assay. In addition, Western blotting was performed to detect the expression of mitogen-activated protein kinase (MAPK) family proteins such as p-extracellular signal-regulated kinase (ERK)1/2, p-p38 and p-c-Jun N-terminal kinase (JNK)1/2, and mitochondrial apoptosis pathway proteins such as B-cell lymphoma (Bcl)-2, Bcl-2 associated X protein (Bax), and cleaved-cysteine aspartic acid specific protease (caspase)-3. RESULTS: Behavioral experiments showed that GP significantly improved the spontaneous activity and motor coordination of PD mice (P<0.05). The forelimb grip strength was also increased by GP treatment (P<0.05), compared to the PD model group. In addition, compared with the model group, the number of TH-positive neurons in substantia nigra pars reticulata region, the levels of dopamine and serotonin in midbrain and the expression of p-ERK1/2 were significantly increased by GP treatment (all P<0.05), whereas the expression of p-p38 and p-JNK1/2, the ratio of Bax/Bcl-2 and cleaved-caspase-3/caspase-3 were significantly decreased (all P<0.05). CONCLUSIONS The results indicate that GP might increase dopamine and serotonin levels in the midbrain and promote the survival of dopaminergic neurons in substantia nigra pars reticulata by regulating the expression of phosphorylation of MAPK family proteins and the expression of mitochondrial apoptosis-related proteins, thereby ameliorating motor deficits in PD mice.
Animals ; Mice ; Male ; Gynostemma/chemistry* ; Mice, Inbred C57BL ; Apoptosis/drug effects* ; Plant Extracts/therapeutic use* ; Parkinson Disease/metabolism* ; Disease Models, Animal ; Neurons/pathology*

Objective:To analyze the relationship between the optimal surgical margin value and clinical prognosis of transoral robotic surgery（TORS） in treating human papillomavirus（HPV） -positive oropharyngeal squamous cell carcinoma. Methods:A single-center, prospective, observational cohort study was conducted, enrolling patients with early and moderated stage（≤T3 stage） oropharyngeal carcinoma undergoing TORS between July 2020 and April 2024. The proposed optimal surgical margin cutoff value for TORS was set as 2 mm. The primary objectives were to evaluate the optimal clear margin for TORS and its association with overall survival（OS） and progression-free survival（PFS）. Logistic regression was used to analyze correlations between surgical margins and clinical variables, while Cox regression models assessed the impact of surgical margins on OS and PFS. Results:A total of 90 patients（60 males, 66.7%） were included, all had squamous cell carcinoma, with a mean age of 58.0±9.0 years（range: 39-84 years） old. The 1, 2 and 3-year OS rates were 92.3%, 89.9% and 85.0%, respectively, while the 1, 2 and 3-year PFS rates were all 90.1%. For surgical margins ≤2 mm, the 1, 2 and 3-year OS rates were 80.8%, 69.3% and 69.3%, respectively, and PFS rates were 77.9% across three time points. For surgical margins>2 mm, the 1, 2 and 3-year OS rates were 96.5%, 96.5% and 90.6%, respectively, with PFS rates of 94.6%. Logistic regression showed no correlation between surgical margins and tumor type, T/N stage, smoking, alcohol use, or gender（P>0.05）. Cox analysis identified surgical margins>2 mm as a significant factor improving PFS（HR=0.14, 95%CI 0.02-0.90, P=0.038）. Conclusion:This systematic analysis suggests setting a 2 mm and longer as clear surgical margin for TORS. Margins>2 mm are associated with superior postoperative PFS rate and prolonged PFS time in HPV-positive oropharyngeal carcinoma patients.
Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Carcinoma, Squamous Cell/virology* ; Human Papillomavirus Viruses/isolation & purification* ; Margins of Excision ; Oropharyngeal Neoplasms/virology* ; Papillomavirus Infections/virology* ; Prognosis ; Prospective Studies ; Robotic Surgical Procedures/methods*

Objective:To investigate the efficacy and safety of venetoclax and azacitidine combined with GHA (human granulocyte colony stimulating factor, homoharringtonine and low-dose cytarabine) priming regimen in treatment of patients with relapsed/refractory acute myeloid leukemia.Methods:A retrospective case series study was conducted. Twenty-three patients with relapsed/refractory acute myeloid leukemia (non-acute promyelocytic leukemia) who received treatment with the combination of venetoclax and azacitidine with GHA priming regimen at the Second Affiliated Hospital of Xi'an Jiaotong University from October 2020 to July 2024 were selected, and the treatment efficacy, minimal residual disease (MRD)-negative rate in patients with comprehensive complete remission (cCR) (including complete remission, complete remission with partial hematologic recovery and complete remission with incomplete hematologic recovery) and the adverse reactions were analyzed; patients were followed-up, and their overall survival (OS) was analyzed by using Kaplan-Meier method.Results:The median age of the 23 patients was 60 years (range: 21-79 years), including 10 males and 13 females. The cCR rate for 1 course of treatment was 52.2% (12/23), with 4 cases of MRD negative among cCR patients; 5 cases received 2 courses of treatment, with 3 cases achieving cCR, of which 2 cases were MRD negative; 2 cases received 3 courses of treatment, with 1 case achieving complete remission with incomplete hematologic recovery. Six patients underwent allogeneic hematopoietic stem cell transplantation. The patients were followed up until July 31, 2024, and the median follow-up period was 5.3 months (range: 1.1-41.7 months). Ten cases survived, 12 cases died, 1 case was lost to follow-up, and the median OS time of 23 patients was 7.9 months. The 6-month OS rate was 60.2% (95% CI: 42.7%-84.8%), and the 12-month OS rate was 44.6% (95% CI: 26.8%-74.3%). Common adverse reactions during treatment included infection [69.6% (16/23)], nausea [56.5% (13/23)], febrile neutropenia [52.2% (12/23)], bleeding [52.2% (12/23)], vomiting [34.8% (8/23)], and pneumonia [34.8% (8/23)]. Conclusions:The combination of vinaclotide and azacitidine with GHA priming regimen has certain efficacy and good safety in the treatment of relapsed/refractory acute myeloid leukemia.

Objective To investigate the correlation between serum adipokine levels and insulin resist-ance in preterm infants with early extrauterine growth retardation(EUGR).Methods A total of 130 preterm infants admitted to the neonatal intensive care unit of the hospital from March 2023 to March 2024 were se-lected as the research subjects.They were divided into the EUGR group(n=63)and the non-EUGR group(n=67)based on whether their weight Z-score at 2 weeks of age was ≥-1.28.Clinical data of the preterm infants and their mothers were collected.Blood samples were collected at 14 days after birth to measure serum insulin,adiponectin,resistin,leptin,blood glucose,total cholesterol,high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),and triglyceride levels.The homeostasis model assessment of insulin resistance(HOMA-IR)was calculated,and the correlation between HOMA-IR and adipokine levels as well as lipid levels was analyzed.Results Compared with the non-EUGR group,the EUGR group had a high-er proportion of mothers with pregnancy-induced hypertension.The preterm infants in the EUGR group had higher levels of insulin,resistin,leptin,triglycerides,and HOMA-IR,and lower levels of HDL-C,with statisti-cally significant differences(P＜0.05).In the EUGR group,HOMA-IR was positively correlated with adi-ponectin,resistin,and leptin levels,while in the non-EUGR group,HOMA-IR was positively correlated with adiponectin levels(P＜0.05).Conclusion Insulin resistance in preterm infants with early EUGR may be re-lated to changes in adipokine levels.

Objective:To screen differentially expressed genes (DEGs) associated with liver cancer by bioinformatics analysis method, and to investigate the mechanism of the minichromosome maintenance protein 2 ( MCM2) and replication factor C subunit 4 ( RFC4) genes in liver cancer in vitro. Methods:Gene expression profiling data of 80 and 36 hepatocellular carcinoma tissues and 307 and 13 cirrhotic tissues were obtained from the GSE25097 and GSE98620 datasets of the gene expression analysis (GEO) database, respectively. Gene expression profiling data of 374 liver cancer tissues and 50 normal liver tissues were downloaded from the cancer genome atlas (TCGA) database. Limma and DESeqs R software were used to process the gene expression profiling data, construct protein-protein interaction networks, and analysis the relevance of these genes to survival. Weighted gene co-expression network analysis was performed to screen out the core genes. Liver cancer SMMC7721 cells were transfected with MCM2 blank plasmid (MCM2 control group), MCM2 overexpression plasmid [MCM2 WT1 group, MCM2 WT2 (2-fold WT1) group], RFC4 blank plasmid (RFC4 control group), and RFC4 overexpression plasmid [RFC4 WT1 group, RFC4 WT2 (2-fold WT1) group], respectively. The expression of MCM2 and RFC4 in liver cancer cell lines and their transfection levels were detected by real-time fluorescence quantitative reverse transcription PCR and Western blotting. The effects of MCM2 and RFC4 on the proliferation of hepatocellular carcinoma cells were detected by MTT assay and cell cloning assay, respectively. The effects of MCM2 and RFC4 on the migration of liver cancer cells were determined by the scratch assay. The effects of MCM2 and RFC4 on liver cancer cell invasion were detected by Transwell assay.Results:By bioinformatic analysis, 9 HCC DEGs were selected, including ubiquitin conjugating enzyme E2 T ( UBE2T), aurora kinase A ( AURKA), targeting protein for Xklp2 ( TPX2), MCM2, RFC4, ribonucleoside-diphosphate reductase subunit M2 ( RRM2), serine peptidase inhibitory factor Kazal type 1 ( SPINK1), collagen type XV alpha 1 chain ( COL15A1) and C-C motif chemokine 25 ( CCL25). Among the six genes associated with clinical stages, the MCM2 and RFC4 genes were found to be strongly associated with prognosis in liver cancer. The relative protein expression of MCM2 and RFC4 in HepG2 cells (1.83±0.07, 1.44±0.09) and SMMC7721 cells (1.74±0.05, 1.43±0.08) was higher than that in MIHA cells (1.00±0.02, 1.00±0.03), and all the differences were statistically significant (all P<0.05). The relative gene expression of MCM2 and RFC4 in HepG2 cells (14.30±0.12, 5.10±0.18) and SMMC7721 cells (10.60±0.11, 7.60±0.07) was higher than that in MIHA cells (1.00±0.05, 1.00±0.03), and all the differences were statistically significant (all P<0.05). Compared with the MCM2 control group, the absorbance values [(0.28±0.01 and 0.21±0.01) vs 0.18±0.03], the number of clonal cells [(717±12 and 782±29) cells vs (389±17) cells], the percentage migration [(0.43±0.02 and 0.68±0.01) vs 0.15±0.06], and the number of cellular invasions [(933±21 and 821±11) cells vs (409±16) cells] were higher in the MCM2 WT1 and MCM2 WT2 groups, and the differences were all statistically significant (all P<0.05). Compared with the RFC4 control group, the absorbance values [(0.30±0.02 and 0.21±0.01) vs 0.17±0.02], the number of cloned cells [(571±11 and 728±9) cells vs (373±23) cells], the percentage migration [(0.75±0.11 and 0.67±0.04) vs 0.34±0.07], and the number of cell invasion [(835±26 and 818±18) cells vs (629±12) cells] were higher in the RFC4 WT1 and the RFC4 WT2 groups, and the differences were statistically significant (all P<0.05). Conclusions:MCM2 and R FC4 genes play a role in promoting tumorigenesis and growth in hepatocellular carcinoma.

Objective:To explore the technical focus of robotic-assisted laparoscopic surgery for the treatment of horseshoe kidney combined with renal tumor.Methods:The clinical data of a patient with horseshoe kidney combined with renal tumor treated by robot-assisted laparoscopic partial nephrectomy in the Second Hospital of Dalian Medical University in September 2021 were retrospectively analyzed. PubMed, CNKI, Wanfang and VIP databases were searched for all the literature on the use of robot-assisted laparoscopic nephrectomy or partial nephrectomy for the treatment of horseshoe kidney combined with renal tumor from the time of establishment to December 2022.Results:A total of 11 patients from 10 articles were retrieved and 12 patients were enrolled. Among the 12 patients, 4 cases used the retroperitoneal approach and 8 cases used the transperitoneal approach. Two cases were operated by traditional laparoscope, and the arteries were searched for and controlled before the robotic arm was placed to perform the partial nephrectomy and suture; and 10 cases were operated with the robotic-assisted laparoscopic approach throughout the whole procedure. Five cases of nephrectomy were performed on one side, and 7 cases were performed in the partial nephrectomy. Postoperative pathological diagnosis was clear cell carcinoma in 8 cases, chromophobe cell carcinoma in 1 case, eosinophilic cell carcinoma in 1 case, renal cell carcinoma in 1 case, and renal abscess in 1 case. The patient in the Second Hospital of Dalian Medical University was 38 years old female who was admitted to the hospital with a fever. After CT arteriography and three-dimensional reconstruction, robotic-assisted laparoscopic partial nephrectomy of right kidney and isthmus dissecting was performed. During the operation, tumor trophoblast vessels were ligated and dissected one by one by using single-use tissue closure clips, and the isthmus was dissected using endoscopic cutting anastomosis on the left side of the tumor, with the tumor edges sharply resected and completely dissected. The operation time was 240 min, without thermal ischemia time, and the bleeding volume was about 300 ml. The patient recovered well after the operation, and the postoperative pathological diagnosis was renal abscess.Conclusions:Robot-assisted laparoscopic treatment of horseshoe kidney combined with renal tumor is safe and effective, and has more advantages than traditional laparoscopic surgery. Preoperative CT arteriography or three-dimensional reconstruction examination should be applied to fully evaluate the variant vessels. The surgical access and plan should be decided according to the size and location of the tumor. The variant vessels should be properly handled during operation. The use of endoscopic cutting anastomosis to deal with the isthmus can be more conducive to the surgical operation.

Objective To explore the protective effect in a model of nicotinamide riboside(NR)against carbonyl cyanide m-chlorophenylhydrazone(CCCP)-induced oxidative stress in R28 cells.Methods 4 μmol/L CCCP was used to induce oxidative stress in R28 cells,and 400 nmol/L NR was used to intervene.The cell viability was quantified by CCK-8 assay.The apoptosis was detected by Annexin-V/PI double staining and flow cytometry.Western blotting was used to examine the levels of Cytochrome C,Caspase-3,and Caspase-9 to evaluate the apoptosis.Tetramethylrhodamine ethyl ester was used to detect the mitochondrial membrane potential(MMP),MitoSOX was used to detect the mitochondrial reactive oxygen species(mtROS)levels,and adenosine triphosphate(ATP)assay kit was used to assess ATP generation ability to evaluate mitochondrial function.Results After CCCP treatment of R28 cells,the cell viability decreased,the apoptotic protein levels and apoptosis rates increased,the MMP decreased,and the mtROS generation increased(P＜0.05).After NR pretreatment,the cell viability increased,the apoptotic protein levels and apoptosis rates decreased,the MMP increased,and the mtROS generation decreased(P＜0.05).Conclusion:NR enhances the cell viability,reduces the expression of apoptotic proteins,and ultimately reduces the apoptosis of retinal ganglion cell by inhibiting oxidative stress response and protecting mitochondrial function.