The seasonal onset or worsening of allergic rhinitis is consistent with the characteristics of not develo-ping， lurking in the body， and recurrent attacks due to the suffering of latent pathogen. Based on the theory of latent pathogen， a seasonal acupuncture treatment approach for allergic rhinitis is proposed. Qi is in the channels in spring， and it is advisable to take the Luo-connecting points and the Ying-spring points and transport points between the muscle interspaces of the body； qi is in the minute collaterals in summer， it is advisable to take the superficial collateral vessels and puncture them for bloodletting； qi is on the skin in autumn， and He-sea points and back-shu points are recommended； qi is in the bone marrow in winter， so it is advisable to take the Jing-well points and Ying-spring points and needle them deeply to the bone.

ObjectiveTo explore the expression levels of long non-coding RNA （lncRNA） H19 and microRNA-22-3p （miR-22-3p） in the serum of patients with acute myocardial infarction （AMI） and their diagnostic value for AMI. MethodsA total of 176 AMI patients were selected as the experimental group and were divided into ST-segment elevation myocardial infarction （STEMI） group （n=95） and non-ST-segment elevation myocardial infarction （NSTEMI） group （n=81） based on their medical history and electrocardiogram. Meanwhile， 156 patients with negative angiography during the same period were selected as the control group （CON group）. The relative expression levels of lncRNA H19 and miR-22-3p in the serum of the two groups were detected by real-time fluorescence quantitative polymerase chain reaction. The diagnostic value of miR-22-3p and lncRNA H19 in AMI was evaluated by receiver operating characteristic curve （ROC） analysis. The levels of serum cardiac troponin I （cTnI）， creatine kinase （CK）， creatine kinase MB isoenzyme （CK-MB）， and high-sensitivity C-reactive protein （CRP） were detected in all study subjects. Spearman correlation analyses were used to evaluate the correlations between lncRNA H19 and miR-22-3p and cTnI， CK， CK-MB and CRP. ResultsCompared with the control group， the expression level of lncRNA H19 was upregulated and the expression level of miR-22-3p was downregulated in the AMI group （all P<0.01）； compared with the NSTEMI group， the expression level of lncRNA H19 was upregulated and the expression level of miR-22-3p was downregulated in the STEMI group （P<0.01）. The areas under the ROC curves （AUC） for the diagnosis of AMI by miR-22-3p， lncRNA H19 and their combination were 0.555， 0.977， and 0.983， respectively. lncRNA H19 was positively correlated with cTnI， CK， CK-MB and CRP （P<0.05）， and negatively correlated with left ventricular ejection fraction （LVEF） （P<0.001）； miR-22-3p was negatively correlated with cTnI， CK and CRP， and positively correlated with LVEF （P<0.05）. ConclusionThe expression level of lncRNA H19 is elevated and the expression level of miR-22-3p decreased in AMI patients， and their levels may have potential value as an auxiliary biomarker for AMI diagnosis and cardiac function assessment.

Objective To investigate the efficacy and safety of fiexible negative pressure suction sheath in the treatment of calyceal diverticulum stones, so as to provide reference for the diagnosis and treatment of this disease.Methods A retrospective analysis was conducted on 5 patients with simple calyceal diverticulum stones treated with flexible ureteroscopy plus fiexible negative pressure suction sheath in our hospital during Jan.2021 and Dec.2024.Data collected included general information, stone location, stone diameter, stone CT value, operation time, postoperative hemoglobin change, and stone free rate.Surgical efficacy and safety were evaluated.Results Among the 5 patients, 4 successfully underwent flexible ureteroscopic lithotripsy, while 1 converted to percutaneous nephrolithotomy due to inaccessible stone location.The median operation time was 120(95-150)min, and the median procalcitonin level was 0.089(0.076-0.112)ng/mL.There was no statistically significant difference in the hemoglobin level between preoperative and postoperative 24-hour measurements(P= 0.075).Abdominal CT scan on postoperative day 1 showed no residual stones, with Clavien grade I ureteral injury in 1 patient. No complications occurred during the 1-month follow-up after surgery.Conclusion Holmium laser lithotripsy under flexible ureteroscopy is one of the common surgical treatment for calyceal diverticulum stones.Combination with fiexible negative pressure suction sheath solves the problem of stone clearance during surgery, reduces the stone residual, demonstrates favorable safety, making flexible ureteroscopy an advantageous approach for treating calyceal diverticulum stones.

Objective To explore the clinical efficacy of robot-assisted coronary artery bypass grafting through a small incision in the left intercostal space in the treatment of multivessel coronary disease. Methods A retrospective analysis was conducted on the clinical data of patients who underwent coronary artery bypass grafting through a small incision in the left intercostal space at Central China Fuwai Hospital of Zhengzhou University from January 1, 2023 to October 15, 2024. Patients were divided into a robotic group and a minimally invasive group based on whether the surgery was assisted by the Da Vinci robot. Results A total of 81 patients were included, with 57 in the minimally invasive group, including 41 males and 16 females, with a median age of 65.0 (57.5, 69.5) years; and 24 in the robotic group, including 17 males and 7 females, with a median age of 61.0 (56.0, 69.0) years. There was no statistically significant difference in baseline data between the two groups (P>0.05). The robotic group had less intraoperative bleeding [300 (200, 438) mL vs. 500 (375, 600) mL, P=0.006], shorter postoperative mechanical ventilation time [15.0 (13.3, 23.5) h vs. 22.0 (15.5, 39.5) h, P=0.037], and lower incidence of postoperative pain [8 (33.3%) vs. 33 (57.9%), P=0.043]. The hospitalization cost in the robotic group was higher than that in the minimally invasive group [130491 (123298, 135691) yuan vs. 123892 (115543, 133449) yuan, P=0.023]. There was no statistical difference in postoperative laboratory indicators between the two groups (P>0.05). There was also no statistical difference in the duration of surgery, postoperative 24 h drainage volume, ICU stay time, postoperative hospital stay or incidences of perioperative compications including pleural effusion, transfusion, new-onset atrial fibrillation, acute kidney injury, non-union of incision, major cardiovascular and cerebrovascular adverse events, and reoperation between the two groups (P>0.05). Conclusion Compared with the minimally invasive group, the robotic group shows satisfactory efficacy and can effectively reduce postoperative pain and intraoperative bleeding, and shorten postoperative mechanical ventilation time.

ObjectiveTo investigate the role and mechanism of ribosomal DNA （rDNA） transcription and ribosome biogenesis in muscle atrophy induced by acute kidney injury （AKI）. MethodsEight male C57BL/6 mice were randomly divided into a control group （Ctrl） and a model group （AKI）. An AKI model was established via intraperitoneal injection of cisplatin. Muscle atrophy was phenotypically assessed by measuring muscle mass， myofiber cross-sectional area （HE staining）， and mRNA expression levels of atrophy-related genes （Murf-1， Atrogin-1， Igf-1） using qRT-PCR. In vivo protein synthesis rates were determined via the SUnSET assay （puromycin incorporation）. Ribosome biogenesis was evaluated by assessing rRNA content and 47S pre-rRNA expression levels. Myotubes differentiated from mouse skeletal muscle cell lines （C2C12 myotubes） were treated with serum from AKI mice， and the effects on rDNA transcription， ribosome biogenesis， and protein metabolism were analyzed using chromatin immunoprecipitation followed by quantitative polymerase chain reaction （ChIP-qPCR） and Western blot. ResultsAKI successfully induced muscle atrophy， as evidenced by a significant reduction in skeletal muscle mass. The most pronounced decrease occurred in the extensor digitorum longus muscle （21.0%， P 0.01）， along with a trend toward reduced myofiber cross-sectional area. At the molecular level， AKI inhibited muscle protein synthesis （83.14% reduction in puromycin incorporation， P 0.000 1） and impaired ribosome biogenesis， manifested by suppressed rDNA transcription elongation （52.62% decrease in 47S pre-rRNA ITS-1 levels， P 0.01） and reduced total rRNA content （65.29%， P 0.000 1）. In contrast， serum from AKI mice promoted rDNA transcription initiation and protein synthesis in C2C12 myotubes in vitro. ConclusionAKI induces muscle atrophy by suppressing rDNA transcription and ribosome biogenesis in skeletal muscle， leading to impaired protein synthesis. Dysregulated ribosome biogenesis may play a critical role in AKI-induced muscle atrophy.

Diarrhea-predominant irritable bowel syndrome （IBS-D） exhibits typical characteristics of psychosomatic comorbidity. Its core pathogenesis is considered to involve simultaneous disorders of both the body and the spirit， for which the former is attributed to liver constraint and spleen deficiency， and dysfunction of large intestine conduction， whereas the latter is induced by spirit restlessness of zang （脏） organs， and gate insecurity of the corporeal soul. Based on this understanding， the therapeutic principle of "simultaneous regulation of body and spirit" is established. For regulating the body， the methods include soothing the liver， fortifying the spleen， consolidating the intestines， and stopping diarrhea， with emphasis on the front-mu （募） points of the related zang-fu （脏腑） organs and acupoints of the foot yangming （阳明） stomach channel and foot taiyin （太阴） spleen channel. For regulating the spirit， the methods include calming the brain spirit， tranquilizing the zang organs spirit， and securing the corporeal soul gate， mainly selecting acupoints of Governer Vessel （督脉） and the back-shu （背俞） points of the corresponding zang-fu organs. Needle manipulation techniques for supplementation and drainage are flexibly applied according to the deficiency or excess nature of the syndrome. In addition， comprehensive application of moxibustion， electroacupuncture， and auricular therapy is emphasized， with the aim of providing clinical insights for diagnosis and treatment.

Purpose: To evaluate the clinical outcomes of endovascular treatment for ruptured vertebral artery dissecting aneurysms (VADAs), with a focus on endovascular protocols tailored to different anatomical subtypes. Materials and Methods: We retrospectively analyzed 44 consecutive patients with ruptured VADAs treated from December 2014 to October 2024. Treatment strategies were guided by aneurysm location: parent artery occlusion (PAO) was used for lesions in non-dominant VAs, while stent-assisted coiling (SAC) reconstruction was used for aneurysms in dominant VAs. Patients were stratified by modified Rankin Scale (mRS) scores at 3-month follow-up (favorable: mRS 0–2; unfavorable: mRS 3–6). Results: All procedures were technically successful, including 12 single SAC, 13 overlapping SAC, and 19 with PAO (2 involving posterior inferior cerebellar artery preservation). Immediate complete occlusion was achieved in 50.0% of single SAC cases, 76.9% overlapping SAC, and 100% in the PAO group. Six (13.6%) procedure-related complications occurred, including 2 hemorrhagic and 4 ischemic events. Thirty patients (68.2%) had favorable outcomes, and the unfavorable outcomes group showed a higher incidence of ischemic events (P=0.012). An initial poor Hunt–Hess grade (≥IV) was an independent risk factor for unfavorable outcomes (odds ratio 6.35, 95% confidence interval 1.24–32.59; P=0.027). Angiographic follow-up was performed for 34 aneurysms, with complete occlusion achieved in 32 (94.1%); the remaining 2 patients were retreated. Conclusion Endovascular management is safe and effective for ruptured VADAs. Anatomical stratification—PAO for non-dominant VA lesions and SAC for dominant VA aneurysms—may help achieve favorable outcomes.

Against the backdrop of the "New Medical Education" initiative by the Ministry of Education, higher medical education faces an urgent need to reconstruct its talent cultivation system. Taking the Laboratory Animal Science course at Shandong University of Traditional Chinese Medicine as an example, this paper systematically explores how to construct a teaching system featuring deep integration of scientific research and education, aiming to address the disconnection between theory and practice and the lack of innovative thinking among students in traditional teaching. The construction of this system is based on three pillars: first, the establishment of a high-level, interdisciplinary faculty team that strongly supports teaching innovation; second, reliance on the Laboratory Animal Center of Shandong University of Traditional Chinese Medicine and the Shandong Provincial Engineering Research Center for Prevention and Treatment of Major Brain Diseases with Traditional Chinese Medicine to provide the necessary facilities; third, a systematic restructuring of teaching content and modes through the "multidisciplinary, multi-format, and multilevel" integration approach. The reform has yielded remarkable results, particularly evident in the increase in students' scientific research and innovation capabilities: the proportion of student projects approved at the provincial level or above in undergraduate innovation training programs significantly increased from 11.1% before the reform to 41.6%. Furthermore, student teams have won gold and silver awards in national competitions such as the "National Undergraduate Innovation Forum for Basic Medical Sciences and Experimental Design" and the "National College Students' Medical Innovation Competition". Additionally, the teaching team's faculty members serving as chief editors and editorial board members of the national planning textbook Laboratory Animal Science published by People's Medical Publishing House mark the nationwide recognition of the course construction achievements. Practice has shown that this integrated system effectively transforms scientific research resources into educational advantages, providing an effective pathway for cultivating top-notch talents in traditional Chinese medicine with solid practical skills and advanced innovative capabilities. It also offers a replicable model for teaching reforms in peer institutions.

CRISPR-Cas-based biosensing systems have become important platforms for rapid nucleic acid detection because of their programmable sequence recognition and high specificity. In many diagnostic workflows, the CRISPR reaction is coupled with polymerase chain reaction or isothermal pre-amplification to enrich low-abundance targets before readout. This design improves sensitivity, but it also introduces additional primers and enzymes, requirements for temperature control and reaction compatibility, and a higher demand for contamination management. These factors make the whole assay less convenient for integrated and field-deployable point-of-care testing. Developing pre-amplification-free CRISPR assays is therefore not simply a pursuit of shorter protocols, but a way to simplify molecular diagnosis at the system level, especially in settings where rapid decisions, closed workflows, and minimal manual operation are required. Microfluidic devices provide an important engineering route for this purpose. Their small reaction volumes, precise fluid manipulation, high-throughput partitioning, and compatibility with portable readout make it possible to integrate target confinement, reaction control, signal acquisition, and quantitative analysis on a chip. Droplet microfluidics and microwell arrays can divide a sample into large numbers of independent microreactors, enabling digital counting of rare recognition events. Continuous-flow, centrifugal, and paper-based microfluidic formats further provide options for automated operation, low-cost fabrication, and on-site use. In this context, microfluidics is not only a signal-enhancement method, but also a platform that connects CRISPR molecular recognition with practical assay implementation. This review summarizes recent progress in microfluidic-based pre-amplification-free CRISPR-Cas biosensing for rapid detection. We focus on four related technical directions. crRNA engineering, including spacer-length tuning, multi-crRNA design, chemical modification, and allosteric regulation, can improve recognition kinetics, nuclease stability, and mismatch discrimination. Digital microfluidic detection based on droplets or microwell arrays converts single-molecule recognition events into countable positive partitions, thereby improving quantitative capability without target pre-amplification. Physical signal transduction interfaces couple CRISPR activity to electrochemical, electrochemiluminescent, fluorescent, or surface-enhanced Raman scattering readouts, making weak molecular signals easier to detect with portable instruments. On-chip cascade signal amplification strategies further enhance output intensity through enzymatic reactions, DNA circuits, nanomaterials, or cross-domain amplification modules while avoiding direct amplification of the target nucleic acid. Together, these strategies show that the performance of a pre-amplification-free assay depends not only on biochemical recognition, but also on how the reaction is confined, amplified, transduced, and operated. In addition to describing these mechanisms, this review compares different strategies in terms of sensitivity improvement, operational complexity, cost, and suitability for point-of-care testing. Representative applications in food authenticity identification and rapid pathogen screening are also discussed to illustrate how these platforms perform in practical scenarios. Finally, we analyze the remaining challenges, including the detection of low-abundance targets in complex matrices, on-chip sample preparation, device-to-device reproducibility, long-term reagent storage, and standardization. Future development will likely depend on tighter integration of sample processing, CRISPR reaction, signal readout, and data interpretation, ultimately moving pre-amplification-free CRISPR diagnostics toward automated and user-friendly sample-to-answer systems.

BACKGROUND:Osteoporotic fracture is the most serious complication of osteoporosis.Previous studies have demonstrated that gut microbiota has a regulatory effect on skeletal tissue and that gut microbiota has an important relationship with osteoporotic fracture,but the causal relationship between the two is unclear. OBJECTIVE:To explore the causal relationship between gut microbiota and osteoporotic fractures using Mendelian randomization method. METHODS:The genome-wide association study(GWAS)datasets of gut microbiota and osteoporotic fracture were obtained from the IEU Open GWAS database and the Finnish database R9,respectively.Using gut microbiota as the exposure factor and osteoporotic fracture as the outcome variable,Mendelian randomization analyses with random-effects inverse variance weighted,MR-Egger regression,weighted median,simple model,and weighted model methods were performed to assess whether there is a causal relationship between gut microbiota and osteoporotic fracture.Sensitivity analyses were performed to test the reliability and robustness of the results.Reverse Mendelian randomization analyses were performed to further validate the causal relationship identified in the forward Mendelian randomization analyses. RESULTS AND CONCLUSION:The results of this Mendelian randomization analysis indicated a causal relationship between gut microbiota and osteoporotic fracture.Elevated abundance of Actinomycetales[odds ratio(OR)=1.562,95%confidence interval(CI):1.027-2.375,P=0.037),Actinomycetaceae(OR=1.561,95%CI:1.027-2.374,P=0.037),Actinomyces(OR=1.544,95%CI:1.130-2.110,P=0.006),Butyricicoccus(OR=1.781,95%CI:1.194-2.657,P=0.005),Coprococcus 2(OR=1.550,95%CI:1.068-2.251,P=0.021),Family ⅩⅢ UCG-001(OR=1.473,95%CI:1.001-2.168,P=0.049),Methanobrevibacter(OR=1.274,95%CI:1.001-1.621,P=0.049),and Roseburia(OR=1.429,95%CI:1.015-2.013,P=0.041)would increase the risk of osteoporotic fractures in patients.Elevated abundance of Bacteroidia(OR=0.660,95%CI:0.455-0.959,P=0.029),Bacteroidales(OR=0.660,95%CI:0.455-0.959,P=0.029),Christensenellacea(OR=0.725,95%CI:0.529-0.995,P=0.047),Ruminococcaceae(OR=0.643,95%CI:0.443-0.933,P=0.020),Enterorhabdus(OR=0.558,95%CI:0.395-0.788,P=0.001),Eubacterium rectale group(OR=0.631,95%CI:0.435-0.916,P=0.016),Lachnospiraceae UCG008(OR=0.738,95%CI:0.546-0.998,P=0.048),and Ruminiclostridium 9(OR=0.492,95%CI:0.324-0.746,P=0.001)would reduce the risk of osteoporotic fractures in patients.We identified 16 gut microbiota associated with osteoporotic fracture by the Mendelian randomization method.That is,using gut microbiota as the exposure factor and osteoporotic fracture as the outcome variable,eight gut microbiota showed positive causal associations with osteoporotic fracture and another eight gut microbiota showed negative causal associations with osteoporotic fracture.The results of this study not only identify new biomarkers for the early prediction of osteoporotic fracture and potential therapeutic targets in clinical practice,but also provide an experimental basis and theoretical basis for the study of improving the occurrence and prognosis of osteoporotic fracture through gut microbiota in bone tissue engineering.