1.Effects and mechanisms of combined exposure to noise and microwave on hippocampal structure and function in mice
Chunxue LU ; Lei SHI ; Yue WANG ; Yanhui HAO ; Xuelong ZHAO ; Yang LI ; Hongyan ZUO ; Liqian ZHU
Journal of Environmental and Occupational Medicine 2026;43(4):419-426
Background Co-exposure to noise and microwave radiation occurs frequently. The central nervous system has been identified as a sensitive target organ for both noise and microwave exposure individually, and the underlying mechanisms remain poorly understood. The specific biological effects resulting from co-exposure to these two factors have yet to be fully elucidated. Objective To clarify the effects of co-exposure to noise and microwave on neurobehavior and hippocampal tissue structure, and to explore the underlying mechanism through the assessment of serum cytokines. Methods C57BL/6N mice were selected and randomly assigned to a blank control group, a noise group, a microwave group, and a combined noise & microwave exposure group. To establish the exposure models, the noise group was subjected to broadband noise at 100 dB for 2 h, while the microwave group received radiation at a central frequency of 9.375 GHz with an average power density of 12 mW·cm−2 and a specific absorption rate of 2.58 W·kg−1 for 15 min. Open field and tail suspension tests assessed anxiety-like emotional behaviour; novel object recognition and Y-maze tests evaluated cognitive function. Histological changes in hippocampal tissue were examined using haematoxylin and eosin (HE) staining, and Nissl staining under light microscopy. Serum cytokine levels were measured using radioimmunoassay and enzyme-linked immunosorbent assay (ELISA). Results After 3 d of exposure, the noise, microwave, and combined exposure groups showed significant reductions in exploration frequency, duration, and distance within the central zone of the open field test compared to the control group (P < 0.01); the combined exposure group exhibited increased ratios of peripheral-to-central exploration time and distance (P < 0.05). After 7 d of exposure, compared with the control group, the noise group maintained a decrease in central zone exploration time (P < 0.01), while the combined exposure group showed persistent decline across all central zone metrics (P < 0.05) and elevated peripheral-to-central ratios (P < 0.05); compared to the microwave group, the combined exposure group showed significant less time in the central zone (P < 0.05) and higher peripheral-to-central ratios (P < 0.05). Regarding behaviour and cognition, compared with the control group, the combined exposure group showed increased immobility time in the tail suspension test after 3 d of exposure (P < 0.01). At this interval, all exposure groups demonstrated reduced frequency and duration of novel object recognition (P < 0.05), with the combined exposure group showing a marked decrease in novel arm exploration time (P < 0.01). After 7 d of exposure, compared with the control group, the noise group showed reduced novel object recognition frequency (P < 0.05), and both the noise and microwave groups exhibited decreased novel arm exploration time (P < 0.05). Pathological alterations including an increased number of hyperchromatic nuclei and depleted Nissl bodies were observed in the CA3 and DG regions across all exposure groups with the most severe lesions observed in the combined exposure group. Serum levels of central nervous system-specific protein β (S-100β), glial fibrillary acidic protein (GFAP), and corticosterone (CORT) were significantly elevated in all exposure groups compared with the control group (P < 0.05). Aquaporin-4 (AQP4) levels increased in the combined exposure group (P < 0.05), while CXC chemokine ligand 10 (CXCL10) levels rose in both the noise and combined groups compared with the control group (P < 0.05). Specifically, S-100β and CXCL10 levels in the combined exposure group were higher than those in the microwave group (P < 0.05); moreover, levels of S-100β, GFAP, CORT, AQP4, and CXCL10 in the combined exposure group were significantly higher than those in the noise group (P < 0.05). Conclusion Combined exposure to noise and microwave radiation induces pathological changes in the hippocampus of mice, increases levels of serum stress hormones and neuro-specific biomarkers. These impairments are more severe than those observed following single-factor exposure. The underlaying mechanism may be related to systemic stress response, neuronal damage, astrocyte activation, and changes in blood-brain barrier permeability, leading to emotional behavioral abnormalities and cognitive decline.
2.Treatment of Liver Cancer by Intervening TGF-β Signaling Pathway with Traditional Chinese Medicine: A Review
Hao CHENG ; Haohao GUO ; Jun SUN ; Juan XUE ; Chunyan JI ; Shiyi LI ; Yuxue DING ; Huaqiang YUE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):318-326
Liver cancer is one of the most common malignant tumors in the digestive system and ranks sixth among newly diagnosed malignant tumors worldwide. Transforming growth factor-β (TGF-β) regulates cell differentiation, proliferation, apoptosis, and other physiological and pathological mechanisms and exerts cancer-suppressive and pro-cancerous dual effects in the process of tumor development. In recent years, with the continuous exploration of the mechanism of liver cancer, it has been found that the conversion of the cancer-suppressive effect into a pro-cancerous effect of this pathway plays a key role in the development of liver cancer. Traditional Chinese medicine (TCM) provides a unique perspective for the classification, diagnosis, and treatment of liver cancer with its comprehensive regulatory effects of multi-components, multi-targets, and multi-pathways. This paper summarized that the cancer-suppressive mechanisms of the TGF-β signaling pathway included promoting cancer cell cycle arrest, apoptosis, autophagy, et al, while the pro-cancerous mechanisms included promoting cancer cell proliferation, invasion and metastasis, immunosuppression, angiogenesis, et al. The TCM compounds intervening this pathway were sorted out, including Jianpi Huayu compound, Fuyang Baoyuan compound, Yipi Yanggan compound, Fuzheng Jiedu compound, compound Astragalus and Salvia, Biejia Jianwan, Dahuang Zhechong pill, and Qingxiang powder. The single TCMs mainly included Schizocapsa plantaginea, Dendrobii Caulis, Gleditsia sinensis, and Dracaena cochinchinensis. The active ingredients of TCM are mainly concentrated on flavonoids, alkaloids, glycosides, phenolics, terpenoids, polysaccharides, and other kinds of compounds. At the same time, it summarized that the liver cancer inhibition mechanism of TCM by regulating this pathway mainly included promoting apoptosis of liver cancer cells, blocking the cell cycle, and inhibiting liver cancer cell proliferation, migration, invasion, angiogenesis, immune escape, etc. The mechanism aims to give full play to the advantages of TCM and precisely regulate the TGF-β signal, thereby exerting positive anti-tumor effects, opening up a new direction for the precise targeted treatment of liver cancer, and providing a scientific basis and a new strategy for the application of TCM in the treatment of liver cancer.
3.Treatment of Liver Cancer by Intervening TGF-β Signaling Pathway with Traditional Chinese Medicine: A Review
Hao CHENG ; Haohao GUO ; Jun SUN ; Juan XUE ; Chunyan JI ; Shiyi LI ; Yuxue DING ; Huaqiang YUE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):318-326
Liver cancer is one of the most common malignant tumors in the digestive system and ranks sixth among newly diagnosed malignant tumors worldwide. Transforming growth factor-β (TGF-β) regulates cell differentiation, proliferation, apoptosis, and other physiological and pathological mechanisms and exerts cancer-suppressive and pro-cancerous dual effects in the process of tumor development. In recent years, with the continuous exploration of the mechanism of liver cancer, it has been found that the conversion of the cancer-suppressive effect into a pro-cancerous effect of this pathway plays a key role in the development of liver cancer. Traditional Chinese medicine (TCM) provides a unique perspective for the classification, diagnosis, and treatment of liver cancer with its comprehensive regulatory effects of multi-components, multi-targets, and multi-pathways. This paper summarized that the cancer-suppressive mechanisms of the TGF-β signaling pathway included promoting cancer cell cycle arrest, apoptosis, autophagy, et al, while the pro-cancerous mechanisms included promoting cancer cell proliferation, invasion and metastasis, immunosuppression, angiogenesis, et al. The TCM compounds intervening this pathway were sorted out, including Jianpi Huayu compound, Fuyang Baoyuan compound, Yipi Yanggan compound, Fuzheng Jiedu compound, compound Astragalus and Salvia, Biejia Jianwan, Dahuang Zhechong pill, and Qingxiang powder. The single TCMs mainly included Schizocapsa plantaginea, Dendrobii Caulis, Gleditsia sinensis, and Dracaena cochinchinensis. The active ingredients of TCM are mainly concentrated on flavonoids, alkaloids, glycosides, phenolics, terpenoids, polysaccharides, and other kinds of compounds. At the same time, it summarized that the liver cancer inhibition mechanism of TCM by regulating this pathway mainly included promoting apoptosis of liver cancer cells, blocking the cell cycle, and inhibiting liver cancer cell proliferation, migration, invasion, angiogenesis, immune escape, etc. The mechanism aims to give full play to the advantages of TCM and precisely regulate the TGF-β signal, thereby exerting positive anti-tumor effects, opening up a new direction for the precise targeted treatment of liver cancer, and providing a scientific basis and a new strategy for the application of TCM in the treatment of liver cancer.
4.Pharmaceutical care for a rare case of diffuse pulmonary microabscesses complicated with organizing pneumonia
Shuangshuang CAI ; Xuewen QIU ; Yue LIAO ; Hongxia ZHAO ; Hao JIANG
China Pharmacy 2026;37(10):1352-1356
OBJECTIVE To investigate the role of clinical pharmacists in the individualized treatment of a rare case of diffuse pulmonary microabscesses caused by Streptococcus constellatus complicated with organizing pneumonia (OP) in a non-immunocompromised patient. METHODS For a middle-aged, non-immunocompromised patient with a one-year disease course, whose imaging findings showed the coexistence of diffuse microabscesses and OP caused by S. constellatus pulmonary abscess, the clinical pharmacist, based on pharmacokinetics/pharmacodynamics characteristics, assisted the physician in optimizing the previously ineffective anti-infective regimen to Ceftriaxone sodium for injection combined with Metronidazole and sodium chloride injection, so as to enhance coverage of mixed anaerobes and lesion penetration. After the patient’s hemoptysis ceased, the clinical pharmacist recommended timely initiation of low-dose Methylprednisolone sodium succinate for injection to manage OP. Upon discharge, sequential oral therapy with Linezolid tablets, Metronidazole tablets, Methylprednisolone tablets and Omeprazole enteric-coated tablets was prescribed. RESULTS After implementation of the individualized treatment regimen, the patient’s hemoptysis resolved, pulmonary lesions significantly regressed, and the patient was discharged in a stable condition. At the three-month follow-up, the patient remained stable. CONCLUSIONS For chronic persistent pulmonary infection complicated with OP in a non-immunocompromised host, clinical pharmacists assist clinicians in optimizing anti-infective regimens and recommend the timely initiation of anti-inflammatory therapy after infection control, thereby contributing to favorable clinical outcomes. Individualized treatment of such complex cases requires comprehensive consideration of pathogen coverage, pathological barriers, and the timing of anti-inflammatory intervention.
5.Overview of the Research on Mechanisms and Application of Essential Oil of Aromatic Chinese Medicinals in Prevention of Respiratory Infectious Disease
Wan Ling LI ; Xinxin WU ; Xiaolei LI ; Mingzhao HAO ; Fang ZHANG ; Yue ZHANG ; Haoyue LI ; Jing ZHAO
Journal of Traditional Chinese Medicine 2025;66(6):638-644
Aromatic Chinese medicinal essential oils are volatile oils extracted from aromatic Chinese herbs, which can prevent and treat respiratory infectious diseases through multiple synergistic mechanisms including pathogen inhibition, immune regulation, and inflammatory response regulation. Essential oils are primarily used externally on the body to prevent infections and alleviate symptoms through methods like inhalation, smearing, topical application, bathing, gargling or as a suppository. They can also be utilized in the environment for disinfection and air purification, through methods like diffusion, vaporization, or spraying. The external application of essential oils extracted from Chinese aromatic herbs has the advantages of convenience, quick absorption, and simultaneous influence on both the body and mind. However, there are still challenges and deficiencies in aspects such as the positioning of functions, indications, safety, and the research on the mechanism of action. It has been proposed to combine the theory of aromatic Chinese medicinals with the characteristics of essential oils, and formulate prescriptions of Chinese medicinal essential oils under the principles of traditional Chinese medicine syndrome differentiation, and prevent and treat respiratory infectious diseases efficiently, accurately, and safely, thereby expanding the clinical application of aromatic Chinese medicinals and the preventive theory of traditional Chinese medicine.
6.Inhibition of excessive inflammatory response of macrophages by Ebselen against acute Escherichia coli infection
Xiao-wen LIU ; Xiao-qin MOU ; Chuang CHENG ; Shuang-shuang GONG ; Hao-ran ZHANG ; Jing HE ; Xi ZHENG ; Jun WANG ; Yue-qing WANG ; Li-li ZOU
Chinese Pharmacological Bulletin 2025;41(7):1346-1353
Aim To investigate the pharmacological mechanism of Ebselenin(Ebselen,EbSe)in the treat-ment of Escherichia coli(E.coli)infection,which had no significant inhibitory effect on Gram-negative bacte-ria,based on previous studies.Methods After EbSe intervention in E.coli infected Raw264.7 cells,the via-bility of Raw264.7 cells was determined by CCK-8 method,the morphology and structure of Raw264.7 cells were observed by electron microscope,and the in-tracellular bacterial load of Raw264.7 cells was calcu-lated by coated plate method.Polarization status of peritoneal macrophages,Raw264.7 intracellular NO and ROS content and intracellular HO-1 expression in Raw264.7 and E.coli acutely infected mice after E.co-li infection by flow cytometry.qPCR was used to detect the expression of related mRNAs in Raw264.7 cells.qPCR was used to detect the intracellular GSH content in Raw264.7 cells by spectrophotometric assay,and the state of cytoskeletal proteins was observed by immuno-fluorescence.Western blot assay was performed to de-tect the intracellular Txnrd1 expression level.Results Microtiter method,CCK-8,and electron microscopy observations showed that EbSe had no effect on the growth of E.coli and Raw264.7 cells in vitro.The re-sults of smear plate counting showed that EbSe reduced the intracellular bacterial load of Raw264.7 in the in-fected group.Flow cytometry results showed that EbSe upregulated the number of M2-type macrophages.The EbSe-treated infected group had reduced intracellular NO and ROS levels and increased GSH levels.The qPCR results showed that the expression of IL-6,IL-1β,and iNOS was decreased,and the expression of HO-1,Txnrd1,and Glut1 was increased in DHB4-in-fected Raw264.7 cells after EbSe treatment.Cytoskel-etal staining showed that the morphology of the EbSe-treated infected cells was similar to that of oxPAPC-in-duced cells.Western blot results showed the expres-sion of Txnrd1 protein in EbSe-treated infected cells in-creased.Conclusion EbSe exerts anti-E.coli acute infection effect by regulating macrophage polarization and inhibiting macrophage excessive inflammatory state.
7.Influence of Oatp4c1-P-gp transmembrane transport system in the kidneys of obese mice on the pharmacokinetics of digoxin
Jing-wen MEN ; Lu SUN ; Hao-ran YUE ; Zhao-yue DUAN ; Hui-nan HOU ; Ting GUI ; Jun-hong XIN ; Zhi-bo GAI
Chinese Pharmacological Bulletin 2025;41(7):1318-1324
Aim To investigate the expression of or-ganic anion transporting polypeptide 4C1(Oatp4c1)-P-glycoprotein(P-gp)in the kidneys of obese mice in-duced by high-fat diet(HFD),and its impact on the pharmacokinetic changes of digoxin.Methods C57BL/6 mice were randomly divided into the Chow group and the HFD group.Body weight and blood glu-cose were recorded weekly.After successful model es-tablishment,digoxin was intraperitoneally injected,and blood was collected at different time points.Part of the blood samples was used for LC-MS/MS detection,and the other part was used for the detection of other bio-chemical indicators.After 16 weeks,the organs were removed and weighed.HE and immunohistochemical staining was used to observe the renal pathology and the expression of Villin,a marker of proximal tubules.Western blot and qPCR were combined to detect the expression of Villin,Oatp4c1 and P-gp.Results In the HFD group,body weight and blood glucose in-creased significantly.The blood concentration of digox-in rose,the area under the curve increased,and the half-life was prolonged.The proximal tubular epithelial cells shed,and the protein expression of Villin,Oatp4c1 and P-gp decreased significantly.Conclu-sions The down-regulation of Oatp4c1-P-gp expres-sion in the kidneys of HFD mice leads to an increase in the blood concentration of digoxin and a decrease in re-nal clearance.
8.Application and prospect of artificial intelligence in the teaching of vascular surgery for overseas trainees
Yue HAO ; Rongjie ZHANG ; Hua ZHONG
Chinese Journal of Medical Education Research 2025;24(2):155-159
As the globalization process accelerates and international medical exchanges become increasingly frequent, the status of foreign trainees in Chinese medical education system has become increasingly important. This paper explores the application of artificial intelligence technology in teaching vascular surgery for foreign trainees, including its specific application forms in theoretical teaching, surgical skill training, case analysis, and clinical decision-making. This article analyzes the advantages of integrating artificial intelligence into teaching foreign trainees, such as improving teaching efficiency, enhancing individualized teaching, and promoting cultural integration. Moreover, this paper discusses the challenges such as data security and privacy protection, technical reliability and adaptability, the transformation of educational concepts, and cross-cultural education, and proposes corresponding strategies to address these issues. The rational use of artificial intelligence technology is expected to improve the teaching quality of vascular surgery for foreign trainees and make a positive contribution to the cultivation of international medical talents.
9.Application value of machine learning prediction model for neural invasion in gallbladder cancer based on enhanced CT and clinical characteristics
Bing ZHOU ; Sheng ZHANG ; Hao LI ; Binjie ZHOU ; Yang JIAO ; Qingwu WU ; Junyan YUE ; Shaoying LI
Chinese Journal of Digestive Surgery 2025;24(4):535-542
Objective:To explore the application value of machine learning prediction model for neural invasion in gallbladder cancer based on enhanced computed tomography (CT) and clinical characteristics.Methods:The retrospective cohort study was conducted. The clinical and imaging data of 502 patients with gallbladder cancer who were admitted to The First Affiliated Hospital of Xinxiang Medical University from January 2010 to June 2024 were collected. There were 171 males and 331 females, aged 65(range, 35?91)years. All patients underwent preoperative abdominal enhanced CT and radical resection. The 502 patients were randomly divided into a training set of 351 cases and a test set of 151 cases at a 7:3 ratio. The training set was used to construct prediction model, and the test set was used to validate prediction model. Observation indicators: (1)neural invasion in gallbladder cancer and influencing factor analysis; (2) construction and validation of machine learning prediction models for neural invasion in gallbladder cancer. Comparison of count data between groups was conducted using the chi-square test. Comparison of ordinal data between groups was conducted using the Mann-Whitney U test. Logistic regression model was performed for univariate and multivariate analyses. Independent influencing factors were incor-porated to construct machine learning models using the standard library modules based on Python 3.9. Receiver operating characteristic (ROC) curves were plotted, and the accuracy, sensitivity, specificity, area under the curve (AUC), precision, F1 score, positive predictive value, negative predic-tive value, and Kappa value were calculated to evaluate the predictive performance of the models. The Delong test was used to assess the differences in AUC among different models in the test set. The Hosmer-Lemeshow test and Brier score were used to evaluate the calibration of the models. Results:(1) Neural invasion in gallbladder cancer and influencing factor analysis. Of the 502 patients with gallbladder cancer, 131 cases had neural invasion, and 371 cases had no neural invasion. Results of multivariate analysis showed that total bilirubin, carcinoembryonic antigen, CA199, CA125, neutrophil-lymphocyte ratio, liver invasion detected by CT, vascular invasion detected by CT, hilar or retroperi-toneal lymph node metastasis detected by CT, and tumor stages T3 and T4 were independent influencing factors for neural invasion in patients with gallbladder cancer [ odds ratios=3.747, 2.395, 3.917, 3.596, 2.805, 2.377, 3.523, 2.774, 5.080, 6.809, 95% confidence interval ( CI) as 1.890?7.430, 1.154?4.971, 2.054?7.472, 1.807?7.155, 1.506?5.225, 1.241?4.553, 1.666?7.449, 1.483?5.189, 2.050?12.589, 2.552?18.168, P<0.05]. (2) Construction and validation of machine learning predic-tion models for neural invasion in gallbladder cancer. Based on the independent influencing factors, seven machine learning models were constructed, including logistic regression, K-nearest neighbors, support vector machine, random forest, decision tree, back-propagation neural network, and gradient boosting machine. The ROC curves of seven machine learning models in the test set were plotted, and the AUC were 0.900(95% CI as 0.851?0.948), 0.741(95% CI as 0.646?0.829), 0.836(95% CI as 0.762?0.895), 0.782(95% CI as 0.701?0.855), 0.839(95% CI as 0.770?0.901), 0.817(95% CI as 0.738?0.887), 0.843(95% CI as 0.770?0.909), respectively. Results of Delong test showed that the logistic regression model had the highest AUC. The sensitivity and specificity of the logistic regression model were 0.868 and 0.805 respectively, indicating the best balance. Results of Hosmer-Lemeshow test showed that the logistic regression model had a good goodness-of-fit ( χ2=5.320, P>0.05). The Brier score of the logistic regression model was relatively low, as 0.168, which verified its calibration advantage. Conclusion:Total bilirubin, carcinoembryonic antigen, CA199, CA125, neutrophil-to-lymphocyte ratio, liver invasion detected by enhanced CT, vascular invasion detected by enhanced CT, hilar or retroperitoneal lymph node metastasis detected by enhanced CT, and tumor stages T3 and T4 are independent influencing factors for nerve invasion in patients with gallbladder cancer. Seven machine learning models are constructed based on enhanced CT and clinical characteristics to predict neural invasion in gallbladder cancer, of which the logistic regression model demonstrates good predictive performance.
10.Comparison and evaluation of three different methods for preparing rat models of lumbar disc herniation
Hao YIN ; Meiqi JI ; Zhixiang HU ; Han WU ; Heng LYU ; Shengyun LI ; Lei LI ; Chuntao ZHAI ; Yue LYU
Chinese Journal of Tissue Engineering Research 2025;29(14):2930-2936
BACKGROUND:Currently,there are various rat models of lumbar disc herniation used in experiments,each with its own advantages and disadvantages.The most common modeling methods include autologous nucleus pulposus transplantation and annulus fibrosus puncture models.OBJECTIVE:To establish two autologous nucleus pulposus transplantation models(with spinous process and mastoid process excision and with transverse process and mastoid process excision)as well as an annulus fibrosus puncture model,and to compare and evaluate the characteristics of the three models.METHODS:Forty male adult Sprague-Dawley rats were randomly divided into four groups(n=10 per group):sham surgery group,spinous process group,transverse process group,and annulus fibrosus puncture group.In the sham surgery group,surgical preparation was performed,the skin was incised,and the spinous process was exposed,and then sutured.In the spinous process group,L5 spinous process and transverse process were excised,and two pieces of tail nucleus pulposus were placed in the intervertebral foramen.In the transverse process group,L5 transverse process and transverse process were excised,and two pieces of tail nucleus pulposus were placed in the intervertebral foramen.In the annulus fibrosus puncture group,the transverse process was excised and annulus fibrosus puncture and intervertebral disc injection of interleukin 1β were then performed.Thermal paw withdrawal latencies were tested before and after modeling.Lumbar spine MRI was performed 2 weeks after modeling.Pathological changes in the intervertebral discs were observed using hematoxylin-eosin staining and safranin-O-fast green staining.Immunofluorescence was used to observe CD68+positive expression.RESULTS AND CONCLUSION:(1)Thermal withdrawal threshold testing results showed that compared with the sham surgery group,pain sensitivity and tolerance time of rats decreased significantly after modeling(P<0.05).(2)Lumbar spine MRI images showed that the spinous process and transverse process groups had obvious protrusion of nucleus pulposus tissue,which more closely resembled MRI images of patients with common lumbar disc herniation.(3)Hematoxylin-eosin staining revealed that compared with the sham surgery group,nucleus pulposus tissues in the model groups showed varying degrees of degeneration,inflammatory cell infiltration,and degradation of spinal cord cells,with the appearance of cystic changes,among which the annulus fibrosus puncture group had the most severe pathological changes.(4)Safranin-O-fast green staining showed that compared with the sham surgery group,the boundaries of nucleus pulposus tissues in the three model groups were blurred,with extensive inflammatory reactions and varying degrees of degeneration in the annulus fibrosus.(5)CD68+immunofluorescence staining results showed that compared with the sham surgery group,the expression of CD68+in the model groups was higher and more widespread,with the annulus fibrosus puncture model showing the highest expression.All the three methods could be used to effectively establish rat models of lumbar disc herniation,with the annulus fibrosus puncture model established after excision of the transverse process being superior to the autologous nucleus pulposus transplantation model(spinous process+mastoid process),and the first two models being superior to the autologous nucleus pulposus transplantation model(transverse process+mastoid process).

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