1.Association of vegetables and fruits intake with depressive symptoms in Yunnan multi ethnic first year junior high school students
CHEN Yiyao*, DU Yeming, YIN Wan, HUANG Jianhong, LIU Zihan, JIANG Yinghong, YU Qiaoying, PAN Lijuan, XU Honglü ;
Chinese Journal of School Health 2026;47(1):32-36
Objective:
To explore the association of vegetables and fruits intake with depressive symptoms among multi ethnic first year junior high school students in Yunnan Province, so as to provide data support for preventing and reducing depressive symptoms among first year junior high school students.
Methods:
From October to December 2022, a cluster random sampling method was used to select 8 500 first year junior high school students from 11 ethnic minority areas in Yunnan Province (Fugong County, Longling County, Longyang District, Luchun County, Mojiang County, Nanjian County, Qiaojia County, Shuangjiang County, Tengchong City, Yuanmou County, Zhenyuan County), to investigate with a questionnaire. The Dietary Frequency Questionnaire was used to collect dietary behavior datas, and the Chinese version of Depression Anxiety Stress Scale-21 (DASS-21) was used to assess depressive symptoms. The generalized linear model was used to analyze the association of vegetable and fruit intake with depressive symptoms in students, and stratified analysis was performed according to ethnicity.
Results:
The detection rate of depressive symptoms among first year junior high school students in Yunnan Province was 29.5%. The detection rates of depressive symptoms in Han and minority first year junior high school students were 26.9% and 31.6%. After controlling for demographic variables such as gender, age, family residence and other confounding factors, the generalized linear model analysis results showed that the intake of leafy vegetables ( β= -0.07 , 95%CI =-0.12 to -0.01), flat fruits ( β=-0.06, 95%CI =-0.12 to -0.00) and hot natured fruits ( β=0.11, 95%CI = 0.04- 0.17) were associated with depressive symptoms in Han first year junior high school students (all P <0.05). The intake of melon and fruit vegetables ( β=-0.06, 95%CI =-0.11 to -0.01) and hot natured fruits ( β=0.06, 95%CI =0.01-0.12) were associated with depressive symptoms in ethnic minority first year junior high school students (both P <0.05).
Conclusions
The intake of vegetables and fruits among multi ethnic first year junior high school students in Yunnan Province is related to the risk of depressive symptoms. It is suggested to strengthen the consumption guidance and education of vegetables and fruits to prevent depressive symptoms among first year junior high school students.
2.Genetic analysis and reproductive intervention for 46 Chinese pedigrees affected with Hereditary multiple exostoses.
Lilan SU ; Xiao HU ; Jing DAI ; Zhengxing WAN ; Duo YI ; Shuangfei LI ; Liang HU ; Yueqiu TAN ; Fei GONG ; Ge LIN ; Guangxiu LU ; Qianjun ZHANG ; Juan DU ; Wenbin HE
Chinese Journal of Medical Genetics 2026;43(4):253-258
OBJECTIVE:
To explore the genetic etiology of 46 Chinese pedigrees affected with Hereditary multiple exostoses (HME) and provide genetic counseling and reproductive intervention.
METHODS:
Whole-exome sequencing and Sanger sequencing were carried out on 87 patients from the 46 pedigrees to analyze the variants of EXT1 and EXT2 genes. Pathogenicity of the variants was assessed based on the guidelines from the American College of Medical Genetics and Genomics and Association for Molecular Pathology (ACMG/AMP). Prenatal diagnosis and preimplantation genetic testing (PGT) were provided for couples with identified pathogenic mutations. This study was approved by the Medical Ethics Committee of the hospital (Ethics No.: LL-SC-SG-2014-010).
RESULTS:
In total 17 and 22 pathogenic variants were respectively identified in the EXT1 and EXT2 genes, among which 5 EXT1 and 12 EXT2 variants were unreported previously. Three patients with no family history were found to harbor de novo variants of the EXT1 gene. Twenty nine couples had opted for PGT or underwent prenatal diagnosis following natural conception, and 17 healthy babies were born.
CONCLUSION
This study has clarified the genetic etiology of 45 HME pedigrees and identified 17 novel variants, which has enriched the mutational spectrum of the EXT1 and EXT2 genes. Reproductive intervention through PGT and prenatal diagnosis have prevented the recurrence of HME in these families.
Humans
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Female
;
Male
;
Pedigree
;
Exostoses, Multiple Hereditary/diagnosis*
;
N-Acetylglucosaminyltransferases/genetics*
;
Adult
;
Exostosin 1
;
Asian People/genetics*
;
Genetic Testing
;
Exostosin 2
;
Mutation
;
China
;
Prenatal Diagnosis
;
Pregnancy
;
Genetic Counseling
;
Preimplantation Diagnosis
;
Exome Sequencing
;
East Asian People
3.Preventive Effect of Intradermal Needle Therapy on Retained Products of Conception after Early Induced Abortion:A Randomized,Single-Blind,Placebo-Controlled Trial
Jun ZHANG ; Pengfei ZENG ; Haihong DU ; Jiao LIU ; Simin WAN
Journal of Traditional Chinese Medicine 2026;67(13):1409-1414
ObjectiveTo evaluate the efficacy and safety of early intradermal needle therapy in preventing retained products of conception (RPOC) following induced abortion. MethodsA randomized, single-blind, placebo-controlled trial design was applied. Eighty-eight patients scheduled for early induced abortion were randomly assigned to either an intradermal needle group (44 cases) or a sham control group (44 cases). Both groups underwent ultrasound-guided vacuum aspiration under general anesthesia and received standard postoperative pharmacological care. Immediately following the operation, the intradermal needle group received active intradermal needle therapy at bilateral Sanyinjiao (SP 6), Hegu (LI 4) and Zhiyin (BL 67), with needles retaining for 3 days, acupoint pressing three times daily for 5 minutes per acupoint, followed by 5 minutes of walking after each pressing session; while the sham control group received a visually identical, blunt-tipped sham needle intervention. The primary outcome was endometrial thickness measured by Doppler ultrasound at week 2 post-procedure. Secondary outcomes included the incidence of RPOC, visual analog scale (VAS) score and PTSD Checklist-Civilian Version (PCL-C) score at 2 hours post-procedure, volume and duration of vaginal bleeding, uterine volume, Pictorial Blood Loss Assessment Chart (PBAC) scores at weeks 5 and 12, menstrual cycle characteristics, and adverse events should be recorded. ResultsAt week 2 post-procedure, endometrial thickness was significantly lower in the intradermal needle group compared to the sham control group (P<0.05). Transvaginal ultrasonography at postoperative week 2 showed thinner endometrium in the intradermal needle group than in the sham group (P<0.05). RPOC occurred 1 case (2.27%) in the intradermal needle group versus 9 cases (20.45%) in the sham control group, yielding a significantly higher RPOC incidence in the sham control group (P<0.05). Patients in the intradermal needle group had less postoperative vaginal bleeding volume and shorter bleeding duration than those in the sham control group (P<0.05). No statistically significant difference was found in 2-hour postoperative VAS pain score, uterine volume at postoperative week 2 and week 5, PBAC score at postoperative week 5 and week 12, or menstrual cycle at postoperative week 12 between groups(all P>0.05). No obvious adverse reactions were observed in either group. ConclusionImmediate intradermal needle intervention after early induced abortion can significantly reduce the incidence of postoperative RPOC, exert beneficial effects on reducing the volume and duration of postoperative vaginal bleeding as well as alleviating post-traumatic stress symptoms, with good safety.
4.Mitochondrial Dysfunction and Diabetic Retinopathy: From Pathogenesis to Therapeutic Targets
Xiao-Yan ZHU ; Tao JIN ; Yu ZHANG ; Lu-Lu LIAN ; Wan-Li DU
Progress in Biochemistry and Biophysics 2026;53(7):1849-1866
Diabetic retinopathy (DR) is one of the most prevalent and vision-threatening microvascular complications of diabetes mellitus, yet its pathogenesis extends far beyond vascular injury alone. As the retina is among the most energy-demanding tissues in the body, its neurons, glial cells, pigment epithelial cells, pericytes, and endothelial cells are highly dependent on mitochondrial oxidative phosphorylation to maintain visual signal transduction, ionic homeostasis, and neurovascular integrity. This review summarizes current evidence indicating that mitochondrial dysfunction is not merely a downstream consequence of chronic hyperglycemia, but a central pathogenic hub that initiates, amplifies, and perpetuates retinal neurovascular degeneration in DR. Persistent hyperglycemia activates multiple abnormal metabolic pathways, including the polyol pathway, hexosamine pathway, protein kinase C signaling, advanced glycation end-product formation, and angiotensin II-related responses. Although these pathways differ mechanistically, they converge on excessive reactive oxygen species (ROS) generation, antioxidant depletion, and mitochondrial injury. Under diabetic stress, electron transport chain overload promotes mitochondrial ROS leakage, damages mitochondrial DNA, disrupts membrane potential, and impairs the transcription of key respiratory chain components. In parallel, mitochondrial quality-control systems become progressively compromised. The balance between fusion and fission shifts toward pathological fragmentation through reduced MFN1/2 and OPA1 activity and enhanced DRP1-mediated fission. Mitochondrial biogenesis is suppressed through inhibition of the AMPK/SIRT1/PGC-1α/NRF1/TFAM axis, while mitophagy changes from an early compensatory response to a later state of autophagic flux blockade and accumulation of dysfunctional mitochondria. Importantly, damaged mitochondria serve as signal amplifiers linking metabolic stress to inflammation and programmed cell death. Mitochondrial ROS, oxidized mitochondrial DNA, calcium overload, cardiolipin exposure, and membrane permeabilization activate interrelated death pathways, including intrinsic apoptosis, ferroptosis, and pyroptosis. Cytochrome C and apoptosis-inducing factor promote caspase-dependent and caspase-independent apoptosis; iron dyshomeostasis, glutathione depletion, GPX4 dysfunction, and lipid peroxidation drive ferroptosis; and mitochondrial danger signals activate the NLRP3 inflammasome and gasdermin-dependent pyroptosis. These pathways jointly damage the retinal neurovascular unit and contribute to pericyte loss, endothelial barrier breakdown, Müller cell dysfunction, retinal ganglion cell apoptosis, retinal pigment epithelial injury, and photoreceptor degeneration. This review also emphasizes the role of epigenetic regulation in stabilizing mitochondrial pathology. DNA methylation, histone modifications, and non-coding RNAs interact to silence mitochondrial protective genes, alter antioxidant responses, and maintain the “metabolic memory” of DR even after glycemic normalization. Therefore, mitochondrial dysfunction should be understood as a dynamic, multidimensional network rather than a single pathological event. Current clinical approaches, such as laser photocoagulation, intravitreal anti-VEGF therapy, and vitrectomy, mainly target advanced vascular lesions and are limited by invasiveness, incomplete responsiveness, recurrence, and potential adverse effects. Therapeutically, strategies targeting mitochondrial ROS, restoring mitochondrial dynamics, enhancing biogenesis, regulating mitophagy, inhibiting inflammasome activation, correcting epigenetic abnormalities, and improving targeted delivery systems show promising potential. However, major translational barriers remain, including retinal cell heterogeneity, stage-specific mitochondrial responses, insufficient organelle-specific drug delivery, and long-term safety concerns. A deeper understanding of mitochondrial regulatory networks may support earlier, more precise, and multi-target interventions for preventing or slowing DR progression.
5.Percutaneous coronary intervention vs . medical therapy in patients on dialysis with coronary artery disease in China.
Enmin XIE ; Yaxin WU ; Zixiang YE ; Yong HE ; Hesong ZENG ; Jianfang LUO ; Mulei CHEN ; Wenyue PANG ; Yanmin XU ; Chuanyu GAO ; Xiaogang GUO ; Lin CAI ; Qingwei JI ; Yining YANG ; Di WU ; Yiqiang YUAN ; Jing WAN ; Yuliang MA ; Jun ZHANG ; Zhimin DU ; Qing YANG ; Jinsong CHENG ; Chunhua DING ; Xiang MA ; Chunlin YIN ; Zeyuan FAN ; Qiang TANG ; Yue LI ; Lihua SUN ; Chengzhi LU ; Jufang CHI ; Zhuhua YAO ; Yanxiang GAO ; Changan YU ; Jingyi REN ; Jingang ZHENG
Chinese Medical Journal 2025;138(3):301-310
BACKGROUND:
The available evidence regarding the benefits of percutaneous coronary intervention (PCI) on patients receiving dialysis with coronary artery disease (CAD) is limited and inconsistent. This study aimed to evaluate the association between PCI and clinical outcomes as compared with medical therapy alone in patients undergoing dialysis with CAD in China.
METHODS:
This multicenter, retrospective study was conducted in 30 tertiary medical centers across 12 provinces in China from January 2015 to June 2021 to include patients on dialysis with CAD. The primary outcome was major adverse cardiovascular events (MACE), defined as a composite of cardiovascular death, non-fatal myocardial infarction, and non-fatal stroke. Secondary outcomes included all-cause death, the individual components of MACE, and Bleeding Academic Research Consortium criteria types 2, 3, or 5 bleeding. Multivariable Cox proportional hazard models were used to assess the association between PCI and outcomes. Inverse probability of treatment weighting (IPTW) and propensity score matching (PSM) were performed to account for potential between-group differences.
RESULTS:
Of the 1146 patients on dialysis with significant CAD, 821 (71.6%) underwent PCI. After a median follow-up of 23.0 months, PCI was associated with a 43.0% significantly lower risk for MACE (33.9% [ n = 278] vs . 43.7% [ n = 142]; adjusted hazards ratio 0.57, 95% confidence interval 0.45-0.71), along with a slightly increased risk for bleeding outcomes that did not reach statistical significance (11.1% vs . 8.3%; adjusted hazards ratio 1.31, 95% confidence interval, 0.82-2.11). Furthermore, PCI was associated with a significant reduction in all-cause and cardiovascular mortalities. Subgroup analysis did not modify the association of PCI with patient outcomes. These primary findings were consistent across IPTW, PSM, and competing risk analyses.
CONCLUSION
This study indicated that PCI in patients on dialysis with CAD was significantly associated with lower MACE and mortality when comparing with those with medical therapy alone, albeit with a slightly increased risk for bleeding events that did not reach statistical significance.
Humans
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Percutaneous Coronary Intervention/methods*
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Male
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Female
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Coronary Artery Disease/drug therapy*
;
Retrospective Studies
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Renal Dialysis/methods*
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Middle Aged
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Aged
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China
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Proportional Hazards Models
;
Treatment Outcome
6.Cell components of tumor microenvironment in lung adenocarcinoma: Promising targets for small-molecule compounds.
Mingyu HAN ; Feng WAN ; Bin XIAO ; Junrong DU ; Cheng PENG ; Fu PENG
Chinese Medical Journal 2025;138(8):905-915
Lung cancer is one of the most lethal tumors in the world with a 5-year overall survival rate of less than 20%, mainly including lung adenocarcinoma (LUAD). Tumor microenvironment (TME) has become a new research focus in the treatment of lung cancer. The TME is heterogeneous in composition and consists of cellular components, growth factors, proteases, and extracellular matrix. The various cellular components exert a different role in apoptosis, metastasis, or proliferation of lung cancer cells through different pathways, thus contributing to the treatment of adenocarcinoma and potentially facilitating novel therapeutic methods. This review summarizes the research progress on different cellular components with cell-cell interactions in the TME of LUAD, along with their corresponding drug candidates, suggesting that targeting cellular components in the TME of LUAD holds great promise for future theraputic development.
Humans
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Tumor Microenvironment/drug effects*
;
Adenocarcinoma of Lung/drug therapy*
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Lung Neoplasms/pathology*
;
Adenocarcinoma/metabolism*
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Animals
;
Apoptosis/physiology*
7.Multi-gene molecular identification and pathogenicity analysis of pathogens causing root rot of Atractylodes lancea in Hubei province.
Tie-Lin WANG ; Yang XU ; Xiu-Fu WAN ; Zhao-Geng LYU ; Bin-Bin YAN ; Yong-Xi DU ; Chuan-Zhi KANG ; Lan-Ping GUO
China Journal of Chinese Materia Medica 2025;50(7):1721-1726
To clarify the species, pathogenicity, and distribution of the pathogens causing the root rot of Atractylodes lancea in Hubei province, the tissue separation method was used to isolate the pathogens from root rot samples in the main planting areas of A. lancea in Hubei. Based on the preliminary identification of the Fusarium genus by the internal transcribed spacer(ITS) sequence, three housekeeping genes, EF1/EF2, Btu-F-FO1/Btu-F-RO1, and FF1/FR1, were amplified and sequenced. Subsequently, a phylogenetic tree was constructed based on these TEF gene sequences to classify the pathogens. The pathogenicity of these strains was determined using the root irrigation method. A total of 194 pathogen strains were isolated using the tissue separation method. Molecular identification using the three housekeeping genes identified the pathogens as F. solani, F. oxysporum, F. commune, F. equiseti, F. tricinctum, F. redolens, F. fujikuroi, F. avenaceum, F. acuminatum, and F. incarnatum. Among them, F. solani and F. oxysporum were the dominant strains, widely distributed in multiple regions, with F. solani accounting for approximately 54% of the total isolated strains and F. oxysporum accounting for approximately 34%. Other strains accounted for a relatively small proportion, totaling approximately 12%. The results of pathogenicity determination showed that there were certain differences in pathogenicity among strains. The analysis of the pathogenicity differentiation of the widely distributed F. solani and F. oxysporum strains revealed that these dominant strains in Hubei were mainly highly pathogenic. This study determined the species, pathogenicity, and distribution of the pathogens causing the root rot of A. lancea in Hubei province. The results provide a scientific basis for further understanding the root rot of A. lancea and its epidemic occurrence and scientifically preventing and controlling this disease.
Plant Diseases/microbiology*
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Atractylodes/microbiology*
;
Phylogeny
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Plant Roots/microbiology*
;
Fusarium/classification*
;
China
;
Virulence
;
Fungal Proteins/genetics*
8.Antithrombotic effect in zebrafish of a fibrinolytic protein EPF3 from Dilong (Pheretima vulgaris Chen) and its transport mechanism in Caco-2 monolayer through cell bypass pathway.
Wan-Ling ZHONG ; Jian-Qiong YANG ; Hai LIU ; Ya-Li WU ; Hui-Juan SHEN ; Peng-Yue LI ; Shou-Ying DU
Journal of Integrative Medicine 2025;23(4):415-428
OBJECTIVE:
EPF3 is a fibrinolysin monomer isolated and purified from Pheretima vulgaris Chen, an earthworm used in traditional Chinese medicine as Dilong for treating blood stasis syndrome. Its composition, anticoagulant and fibrinolytic activities, and relevant mechanisms have been confirmed through in vitro experiments. However, whether it has antithrombotic effects in vivo and can be absorbed by the gastrointestinal tract is unknown. This study evaluates the antithrombotic effect in zebrafish and investigates the gastrointestinal stability and intestinal absorption mechanism of this protein in vitro.
METHODS:
The antithrombotic effect of EPF3 in vivo was verified using the zebrafish thrombus model induced by arachidonic acid and FeCl3. Then, the protein bands of EPF3 incubated with simulated gastric fluid (SGF), simulated intestinal fluid (SIF), and homogenate of Caco-2 cells (HC2C) were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to evaluate its gastrointestinal stability. Finally, the transport behavior and absorption mechanism of EPF3 were studied using Caco-2 cell monolayer.
RESULTS:
EPF3 could significantly enhance the returned blood volume and blood flow velocity in zebrafish with platelet aggregation thrombus induced by arachidonic acid. It could also prolong the formation time of tail artery thrombus and increase the blood flow velocity in zebrafish with vessel injury thrombus induced by FeCl3. EPF3 was stable in SIF and HC2C and unstable in SGF. The permeability of EPF3 in Caco-2 monolayer was time-dependent and concentration-dependent. The efflux ratio was less than 1.2 during transport, and the transport behavior was not affected by inhibitors. EPF3 could reversibly reduce the expression of tight junction-related proteins, including zonula occludens-1, occludin, and claudin-1 in Caco-2 cells.
CONCLUSION
EPF3 could play a thrombolytic and antithrombotic role in zebrafish. It could be transported and absorbed into the intestine through cellular bypass pathway by opening the intestinal epithelium tight junction. This study provides a scientific explanation for the antithrombotic effect of earthworm and provides a basis for the feasibility of subsequent development of EPF3 as an antithrombotic enteric-soluble preparation. Please cite this article as: Zhong WL, Yang JQ, Liu H, Wu YL, Shen HJ, Li PY, Du SY. Antithrombotic effect in zebrafish of a fibrinolytic protein EPF3 from Dilong (Pheretima vulgaris Chen) and its transport mechanism in Caco-2 monolayer through cell bypass pathway. J Integr Med. 2025; 23(4): 415-428.
Animals
;
Zebrafish
;
Humans
;
Caco-2 Cells
;
Fibrinolytic Agents/pharmacology*
;
Thrombosis/drug therapy*
;
Intestinal Absorption
9.The reduction of reproductive function of male zebrafish caused by high-fat diet:A molecular basis study
Zhenghong DU ; Fei MAO ; Xue ZHAO ; Shuo WANG ; Yongjun WAN ; Chengyu ZHANG ; Peng DUAN ; Dahu ZHANG
National Journal of Andrology 2025;31(11):963-971
Objective To investigate the effects and mechanisms of high-fat diet(HFD)-induced obesity on male ze-brafish reproductive function.Methods Adult male zebrafish were divided into normal diet(ND)group and HFD group.Growth and metabolic conditions were evaluated by measuring body weight,body length,BMI,organ index,and glucose/lipid lev-els.Reproductive capacity was assessed via sperm concentration,motility,fertilization rate,and testosterone levels.Testicular tissues from both of groups were subjected to transeriptomic sequencing(RNA-seq).And qRT-PCR was used to validate the expression of genes.Results Compared to male zebrafish in ND group,the ones in HFD group exhibited hepatic steatosis and glucose/lipid meta-bolic disorders(P<0.05).Testicular structural disorganization,along with reduced testosterone levels,decreased gonadosomatic in-dex,and impaired sperm concentration and motility occurred in HFD group(P<0.05).GO analysis revealed that differentially ex-pressed genes were enriched in spermatogenesis and ciliary system,while KEGG analysis highlighted metabolic related pathways(pu-rine metabolism,thyroid hormone synthesis,mTOR signaling)and cell adhesion molecules.Twenty key differentially expressed genes were validated by qRT-PCR,which confirmed the reliability of RNA-Seq results.Conclusion Impairment of reproductive function induced by HFD in zebrafish may be associated with three regulatory mechanisms including ciliary system,metabolic dysregulation,and aberrant cell adhesion molecule signaling.This study provides mechanistic insights and identifies potential therapeutic targets for clinical management of diet-associated infertility.
10.Correlation between poor position of lumbar pedicle screws and postoperative neurological symptoms
Wen WAN ; Weicheng CHEN ; Weiwen CHEN ; Ning ZHANG ; Liuxue DU ; Jiangwei CHEN ; Rongping ZHOU ; Zhili LIU ; Shanhu HUANG ; Jiaming LIU
The Journal of Practical Medicine 2025;41(20):3220-3227
Objective To analyze the correlation between the poor position of pedicle screw after lumbar internal fixation and the occurrence of postoperative neurological symptoms.Methods The clinical data of patients who underwent lumbar pedicle screw surgery in our hospital from January 2017 to December 2023 were analyzed retrospectively.Record the patient's sex,age,diagnosis,surgical segment,the presence of postoperative neurological complications,the types of neurological complications.According to the postoperative CT,the screw penetration zone and grade were evaluated,and the distribution of different pedicle screw penetration types was recorded.The correlation between different types of pedicle screw penetration and postoperative neurological symptoms was analyzed.Results A total of 184 patients were included,including 80 males and 104 females.Age ranged from 18 to 82 years,with an average of 60.1±14.2 years.35 patients developed new neurological symptoms after operation.The incidence of postoperative neurological symptoms caused by pedicle medial quadrant screw penetration and lower quadrant screw penetration was higher than that of pedicle superior quadrant and lateral quadrant screw penetration.From L1 to L5,the risk of postoperative neurological symptoms caused by screw penetration in the medial quadrant of the pedicle increases gradually.When the penetration distance of pedicle medial quadrant screws in L4 and L5 segments is≥2mm,the risk of postoperative neurological symptoms is high.When the penetration distance of pedicle medial quadrant screws in L1,L2 and L3 segments is greater than 4mm,the risk of postoperative neurological symptoms is high.The distance of pedicle screw penetrating the medial quadrant of pedicle was positively correlated with the incidence of postoperative neurological symptoms(P<0.05).Conclusion The lateral and superior quadrants of the pedicle may be regarded as safety zones for lumbar pedicle screw placement.For the L1~L3 segments,screw penetration depth should not exceed 4 mm;whereas for the L4~L5 segments,the safety threshold must be strictly controlled within 2 mm.


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