ObjectiveTo clarify whether METTL14 mediates the core role of acupuncture at Neiguan (PC6) in promoting myelination and improving behavior in young autistic rats through gene intervention technology. MethodsThe ASD model was established by intraperitoneal injection of valproic acid (VPA) in pregnant rats. Male offspring were intracerebroventricularly injected with adenovirus-packaged METTL14 shRNA (sh-METTL14) or its control (sh-NC) on postnatal day 1, with a model group set as well. Subsequently, the juvenile rats were divided into model group, acupuncture group, acupuncture+sh-NC group, and acupuncture+sh-METTL14 group. The acupuncture group received acupuncture at Neiguan (PC6) from postnatal day 7, once daily for 21 consecutive days. Neurobehavioral changes were evaluated by behavioral tests; METTL14 knockdown efficiency and the expression of METTL14, METTL3, and PTEN were detected by quantitative real-time PCR (qRT-PCR) and Western blot (WB); PTEN m⁶A levels were measured by RNA immunoprecipitation-qPCR (RIP-qPCR); myelin ultrastructure, expression of myelin basic protein (MBP) and neurofascin 155 (NF155), and dendritic spine density were observed using transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, qRT-PCR, and primary neuron culture. ResultsBehaviorally, knockdown of METTL14 significantly counteracted the beneficial effects of acupuncture in improving self-grooming, open field exploration, three-chamber social interaction, and Morris water maze learning and memory (P<0.05, P<0.01). Compared with the acupuncture+sh-NC group, the acupuncture+sh-METTL14 group showed significantly decreased mRNA and protein expression of hippocampal METTL14 (P<0.01), and the upregulating effects of acupuncture on METTL3 and PTEN expression were reversed (P<0.01). Meanwhile, knockdown of METTL14 significantly inhibited the acupuncture-induced increase in PTEN m⁶A levels (P<0.01). Morphologically, knockdown of METTL14 attenuated the improvement of myelin structure by acupuncture, reversed the downregulation of MBP and upregulation of NF155 induced by acupuncture, and blocked the increase in dendritic spine density (P<0.05, P<0.01). ConclusionMETTL14 is a key molecule mediating the therapeutic effect of acupuncture at Neiguan. Acupuncture at Neiguan upregulates METTL14, thereby enhancing m⁶A methylation modification of PTEN mRNA to stabilize its expression, ultimately promoting myelin development and improving behavioral symptoms in ASD juvenile rats. This preliminarily reveals the modern biological connotation of “opening Xuanfu and dredging myelin”.

ObjectiveTo clarify whether METTL14 mediates the core role of acupuncture at Neiguan (PC6) in promoting myelination and improving behavior in young autistic rats through gene intervention technology. MethodsThe ASD model was established by intraperitoneal injection of valproic acid (VPA) in pregnant rats. Male offspring were intracerebroventricularly injected with adenovirus-packaged METTL14 shRNA (sh-METTL14) or its control (sh-NC) on postnatal day 1, with a model group set as well. Subsequently, the juvenile rats were divided into model group, acupuncture group, acupuncture+sh-NC group, and acupuncture+sh-METTL14 group. The acupuncture group received acupuncture at Neiguan (PC6) from postnatal day 7, once daily for 21 consecutive days. Neurobehavioral changes were evaluated by behavioral tests; METTL14 knockdown efficiency and the expression of METTL14, METTL3, and PTEN were detected by quantitative real-time PCR (qRT-PCR) and Western blot (WB); PTEN m⁶A levels were measured by RNA immunoprecipitation-qPCR (RIP-qPCR); myelin ultrastructure, expression of myelin basic protein (MBP) and neurofascin 155 (NF155), and dendritic spine density were observed using transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, qRT-PCR, and primary neuron culture. ResultsBehaviorally, knockdown of METTL14 significantly counteracted the beneficial effects of acupuncture in improving self-grooming, open field exploration, three-chamber social interaction, and Morris water maze learning and memory (P<0.05, P<0.01). Compared with the acupuncture+sh-NC group, the acupuncture+sh-METTL14 group showed significantly decreased mRNA and protein expression of hippocampal METTL14 (P<0.01), and the upregulating effects of acupuncture on METTL3 and PTEN expression were reversed (P<0.01). Meanwhile, knockdown of METTL14 significantly inhibited the acupuncture-induced increase in PTEN m⁶A levels (P<0.01). Morphologically, knockdown of METTL14 attenuated the improvement of myelin structure by acupuncture, reversed the downregulation of MBP and upregulation of NF155 induced by acupuncture, and blocked the increase in dendritic spine density (P<0.05, P<0.01). ConclusionMETTL14 is a key molecule mediating the therapeutic effect of acupuncture at Neiguan. Acupuncture at Neiguan upregulates METTL14, thereby enhancing m⁶A methylation modification of PTEN mRNA to stabilize its expression, ultimately promoting myelin development and improving behavioral symptoms in ASD juvenile rats. This preliminarily reveals the modern biological connotation of “opening Xuanfu and dredging myelin”.

ObjectiveThis paper aims to conduct a secondary analysis of a randomized controlled trial on the treatment of myelodysplastic syndrome （MDS） with deficiency of both the spleen and kidney and blockage of toxin and blood stasis with an arsenic-containing traditional Chinese medicine compound， by applying the mixed model for repeated measure （MMRM） and the method of stratified composite outcome with win ratio. The analysis includes the assessment of hematological efficacy and the composite outcome evaluation of adverse reactions， so as to more comprehensively assess the therapy of this regimen. MethodsThe MMRM and win ratio methods were used to evaluate the efficacy of a prospective，multi-center，double-blind，randomized controlled study. The blood routine （hemoglobin concentration，neutrophil count， and platelet count） and biochemical indexes （aspartate aminotransferase，alanine aminotransferase，serum creatinine，and serum ferritin） of the patients were detected at the time of enrollment and at the end of each course of treatment in the laboratory department of Xiyuan Hospital. The patients' syndromes at the time of enrollment and after treatment were recorded and scored according to the therapy standard of traditional Chinese medicine for diseases and syndromes. MMRM was used to analyze the blood routine indexes of the experimental group and the control group. This method has the advantages of high data reliability and dynamic efficacy under intervention and time. The win ratio method was used to evaluate the composite outcome of traditional Chinese medicine syndrome scores and biochemical indexes according to the priority and to verify the clinical safety of arsenic-containing traditional Chinese medicine compound. ResultsThe results of MMRM analysis showed that the hemoglobin concentration of patients in the group with arsenic-containing traditional Chinese medicine compound increased significantly compared with that before treatment in the group，while that in the placebo group decreased significantly （P<0.01）. When compared with that after treatment in the placebo group，the hemoglobin concentration of patients in the group with arsenic-containing traditional Chinese medicine compound increased significantly，and the mean difference of least squares （LS） was statistically significant （P<0.01）. When compared with those before treatment in the group，there were no statistically significant differences in the neutrophil count and platelet count in both groups. After treatment，there were no statistically significant differences in the neutrophil count， platelet count， and the mean difference of LS between the two groups. The analysis results of win ratio showed that the group with arsenic-containing traditional Chinese medicine compound had a significant advantage in the comparison of composite outcomes，with a win ratio （95% CI） of 2.01 （1.24-3.27） （P<0.01），and that the possibility of "winning" in terms of safety was 2.01 times that of the placebo group. The safety advantage of the group with arsenic-containing traditional Chinese medicine compound mainly came from the traditional Chinese medicine syndrome scores，renal function indexes， and iron reserve capacity indexes，and the number of winning times was less than that of losing times in the comparison of liver function outcomes. ConclusionThe MMRM analysis proves that the arsenic-containing traditional Chinese medicine compound can significantly improve the hemoglobin concentration of patients with myelodysplastic syndrome with refractory cytopenia and multilineage dysplasia （MDS-RCMD） of the type of deficiency of both the spleen and kidney and blockage of toxin and blood stasis. This conclusion is not interfered with by time trends and individual relationships and methodologically improves the credibility of the therapy of the arsenic-containing traditional Chinese medicine compound in treating MDS. Four outcomes are evaluated by the win ratio method，namely traditional Chinese medicine syndromes，liver function，renal function， and iron reserve capacity，proving that the arsenic-containing traditional Chinese medicine compound has the comprehensive advantages of improving the survival quality of the patients and reducing adverse reactions. The win ratio outcome provides clear comparative indexes for the evaluation of adverse reactions，making it easier for regulatory authorities，medical staff， and patients to understand the safety of the arsenic-containing traditional Chinese medicine compound in clinical application.

Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.

The pathogenesis of recurrent acute exacerbations of bronchiectasis is considered to be "yang floating and yin weakness". Yin weakness refers to the depletion of the water and fire of true yin in the lower jiao （焦）， which is the root of the disease. Yang floating refers to the ministerial fire floating upward and scorching the lung metal due to the failure of true yin to nourish and constrain yang， which is the manifestation of acute exacerbations. Yin weakness and yang floating interact with each other， forming a vicious cycle of "yin depletion leading to fire floating， and floating fire in turn injuring yin"， which leads to recurrent， prolonged， and refractory acute exacerbations of bronchiectasis. Based on the pathogenesis of yang floating and yin weakness， therapeutic formulas are proposed. Yinhuo Decoction （引火汤） can be used to nourish true yin and anchor floating fire， while a combination of Qianyang Pill （潜阳丹） and Fengsui Pill （封髓丹） is applied to warm the kidneys， subdue yang， and secure true fire， and Quanzhen Yiqi Decoction （全真一气汤） is utilized to drastically supplement the primal origin and rescue both yin and yang simultaneously.

Objective To investigate changes in the distribution of Oncomelania hupensis snails in Yixing City, Jiangsu Province, from 2008 to 2024, and to provide insights to guide formulation of O. hupensis snail investigation and surveillance programs. Methods Data from O. hupensis snail surveys in Yixing City from 2008 to 2024 were collected. Trends in snail habitat site numbers, snail-infested areas, proportion of total area with snail habitats, occurrence of frames with living snails, and density of living snails in snail habitats in hilly regions of Yixing City were evaluated from 2008 to 2024 by applying a Joinpoint regression model to collect annual percent change (APC) and average annual percent change (AAPC) data. Results From 2008 to 2024, a total of 193.815 hm2 of snail-infested area was found within Yixing City, of which 191.765 hm2 (accounting for 98.94%) was distributed in hilly regions. A total of 344 snail infestation sites in hilly regions were identified. The average occurrence rate of living snail frames in snail habitats was 8.650% and the average living snail density in snail habitats was 0.246 per 0.1 m2. No Oncomelania snails were positively identified. The number of snail habitat sites in hilly regions of Yixing City showed no significant decreasing trend (AAPC=-2.8%, P=0.527). Similarly, no significant increasing trend in snail-infested area (APC=AAPC=5.7%, P=0.328) was found. The occurrence rate of living snail frames in snail habitats showed no significant increasing trend (APC=AAPC=5.1%, P=0.583). Living snail density in snail habitats also showed no significant increasing trend (APC=AAPC=3.2%, P=0.598). Conclusions From 2008 to 2024, snail infestation has persisted in the hilly areas of Yixing City with an unclear variation trend. The quality of snail investigation and snail surveillance system should be strengthened to permit timely identification of hidden dangers and rapid risk responses, thereby preventing schistosomiasis outbreaks.

Objective To evaluate the safety and efficacy of a new domestic ultra-high frequency bipolar radiofrequency ablation device. Methods Six fresh cadaveric lumbar spine specimens were used as experimental subjects, and a novel domestic ultra-high frequency bipolar radiofrequency ablation device was employed for nucleus pulposus ablation and annulus fibrosus cauterization. Nucleus pulposus ablation: a total of 18 intervertebral discs from L₁ to L₄ were selected and randomly divided into six groups. Radiofrequency electrodes were inserted into the intervertebral discs through the lateral aspect of the intervertebral foramen, and the ablation was performed at power outputs of 50 W, 75 W, and 100 W for 1 min and 2 min, respectively. The ablation of the nucleus pulposus was observed, and the ablation range was recorded. Annulus fibrosus cauterization: a total of 12 intervertebral discs from L₄ to S₁ were divided into three groups based on cauterization duration of 3 s, 10 s, or 15 s. In a water bath maintained at a constant 37 °C, cauterization was performed at power outputs of 20 W, 30 W, and 40 W, respectively. After hard tissue sectioning and hematoxylin and eosin (HE) staining, the depth of cauterization and the carbonization of surrounding soft tissues were observed. Results No obvious vaporization cavity was observed in the nucleus pulposus at a power of 50 W for either 1 or 2 min. The 100 W–2 min group achieved the largest ablation area ([7.471±0.365] mm², P＜0.001). The temperature elevation at all four annulus fibrosus sites remained below 0.5℃ throughout the ablation process. The cauterization depth in the 20 W group was significantly lower than that in both the 30 W and 40 W groups across all time conditions (P＜0.01), whereas no statistically significant difference was observed between the latter two. In 20 W group, the cauterization depth at 3 s was significantly lower than that of the 10 s and 15 s groups (P＜0.01). No significant intra-group differences were found among subgroups with different cauterization durations in the 30 W and 40 W groups. An effective cauterization depth was already achieved at 30 W for 3 s. Conclusions Ex vivo experiments have shown that the new domestic bipolar radiofrequency ablation device possesses excellent ablation efficacy and safety parameters. For nucleus pulposus ablation, it is recommended to use 100 W for 2 min, while for annulus fibrosus cauterization and shaping, 30 W for 3 s is suggested. Its clinical application value remains to be further verified through in vivo studies.

OBJECTIVE To investigate the effectiveness of different bacterial cleaning methods and their effects on the general condition,nasal mucosal and systemic inflammation of mice.METHODS A total of 44 mice were randomly divided into six groups:4-antibiotic per os group(4ABX po)with 7 mice,5-antibiotic oral gavage group(5ABX og)with 8 mice,5-antibiotic intranasal group(5ABX in)with 8 mice,and each of their control groups with 7 mice.Body weight,water intake,and peripheral blood routine test of mice were measured.Bacterial culture of nasal lavage fluid(NLF)was performed;mRNA level of inflammatory mediators and histopathological analysis were conducted with mouse nasal mucosa.RESULTS Bacteria were cultured from all control groups,while Bacteria were cultured from all control groups,while one mouse in the 4ABX po group,three mice in the 5ABX og group,and two mice in the 5ABX in group showed no bacterial growth.The number of goblet cells in the nasal mucosa significantly increased in the 5ABX og group compared with its control group(P<0.05).The 5ABX in group exhibited significantly higher counts of peripheral blood lymphocytes and hemoglobin levels,as well as greater nasal mucosal thickness compared with its control group,with a notable decrease in goblet cells(P<0.05).No statistical differences were observed in body weight or the mRNA expression of nasal mucosal inflammatory mediators.CONCLUSION Different combinations of antibiotics and administration routes have varying effects on nasal bacteria,systemic and nasal mucosal inflammation in mice.Therefore,choosing appropriate protocols is crucial for the progression of subsequent research.

BACKGROUND:Existing studies have compared patients with degenerative lumbar spondylolisthesis with healthy populations,but comparative studies focusing on differences in this aspect of the parameter between these different lumbar spondylolisthesis are lacking.OBJECTIVE:To explore the differences in paraspinal muscle degeneration between isthmic spondylolisthesis and degenerative lumbar spondylolisthesis,and their correlation with low back pain symptoms.METHODS:A total of 107 patients with lumbar spondylolisthesis treated in Affiliated Hospital of Xuzhou Medical University between February 2019 and August 2023 were analyzed retrospectively.They were divided into the isthmic spondylolisthesis group(39 patients)and the degenerative lumbar spondylolisthesis group(68 patients)according to the type of spondylolisthesis.Paravertebral muscle tissue parameters were compared and analyzed as well as functional scores(visual analog scale for low back pain)in both groups.According to the visual analog scale score,the two groups of patients were divided into two subgroups:those with a visual analog scale score＜45 mm and those with a visual analog scale score＞45 mm,and the relationship between the difference in visual analog scale score and the parameters of paraspinal muscle tissue degeneration was analyzed.RESULTS AND CONCLUSION:(1)The percentage of fat infiltration in the multifidus muscle was higher in the degenerative lumbar spondylolisthesis group than in the isthmic spondylolisthesis group(P=0.003).(2)The percentage of fat infiltration in the multifidus muscle was significantly lower in patients with a visual analog scale score of＜45 mm than in patients with a visual analog scale score of＞45 mm in both groups(P=0.021,P＜0.001).(3)Patients in the isthmic spondylolisthesis group also showed a significantly lower percentage of fat infiltration in patients with visual analog scale score＜45 mm compared with those with visual analog scale score＞45 mm(P=0.002).(4)These results showed that there was a significant difference in paraspinal muscle tissue degeneration between isthmic spondylolisthesis patients and degenerative lumbar spondylolisthesis patients,and that degenerative lumbar spondylolisthesis patients demonstrated more severe paraspinal muscle degeneration compared to isthmic spondylolisthesis patients.In addition,the percentage of fatty infiltration of the multifidus muscle was relatively greater in those with higher pain scores in both groups of lumbar spondylolisthesis patients.

Objective To evaluate the diagnostic efficacy of the Kaiser score for ultrasound(US)BI-RADS 4A lesions and the potential to avoid unnecessary biopsies.Methods Data of patients who underwent breast US and magnetic resonance imaging(MRI)between May 2019 and June 2023 were retrospectively collected,and all cases were confirmed by puncture biopsy or surgical pathology.The diagnostic efficacy of Kaiser score and MRI BI-RADS classification for US BI-RADS 4A lesions was calculated and compared using pathologic findings as the gold standard.Results A total of 119 lesions(74 benign and 45 malignant lesions)were found in the 119 patients.The diagnostic efficacy of the Kaiser score was significantly better than that of the MRI BI-RADS classification(AUC:0.88 vs.0.65,P＜0.000 1).The concordance of the Kaiser score results with the pathologic findings and the inter-observer concordance were better than that of the MRI BI-RADS classification.Unnecessary biopsies could be avoided in 87.8％(65/74)and 33.8％(25/74)of the cases when the Kaiser score and the MRI BI-RADS classification were applied for the diagnosis of US BI-RADS 4A lesions,respectively.Conclusion The Kaiser score can provide a diagnostic basis for risk stratification of US BI-RADS 4A lesions,and its diagnostic efficacy and inter-observer agreement are superior to those of MRI BI-RADS classification.