Objective To explore whether creatine therapy regulates neuronal ferroptosis by inhibiting the activation of STAT1 signaling pathway associated with suppressor of cytokine signaling 1 (SOCS1) in Alzheimer’s disease.   Methods  Immunohistochemical staining and counting of positive results using paraffin sections of human brain frontal lobes were employed to determine the trend of changes in the target proteins. Further validation was performed by immunofluorescence and Western blotting. STAT1 phosphorylation was inhibited by creatine injection using eleven FAD4Tmice and by cerebellar medullary pool puncture, and the expression of target proteins was examined by immunohistochemistry and immunofluorescence after postmortem sampling.   Results  Compared with the age controls, interferon-γ (IFN-γ), an activating cytokine of the STAT1 signaling pathway, and SOCS1, a negative regulator of STAT1 activation, were both significantly up-regulated, STAT1 phosphorylation was enhanced, and the ferroptosis markers ferritin light chain (FTL) and cystine/glutamate transporter(xCT) increased markedly in the cortex of AD human brains; Creatine treatment of FAD4Tmice resulted in a reduction of both IFN-γ and SOCS1, and a significant decrease in the ferroptosis markers FTL and xCT (SLC7A11).  Conclusion  Creatine ameliorates neuronal ferroptosis in AD model mice by reducing neuronal STAT1SOCS1 signalling activation. 

Objective  To explore the neuroprotective effects and mechanisms of the sesquiterpene lactone compound ACT001 on rotenone (ROT) -induced Parkinson’s disease (PD) model mouse.  Methods  SPF C57BL/6 mice were randomly divided into 6 groups, including control group, solvent control group, ROT model group, ACT001 5 mg/kg group（ROT+ACT001-5）, ACT001 20 mg/kg group（ROT+ACT001-20）, and levodopa(L-dopa) positive control group(ROT+L-dopa)，with 9 mice in each group. The control group received an equivalent amount of intraperitoneal injection of saline, the solvent control group received an equivalent amount of rotenone solvent without rotenone, the remaining groups of mice were used to establish a PD mouse model by intraperitoneal injection of rotenone. Mice in different ACT001 dosage groups received intraperitoneal injections of high and low doses of ACT001, while the positive control group received levodopa intraperitoneally for 15 consecutive days. Behavioral changes in mice were assessed using open field, rotarod, pole-climbing, and balance beam tests. Immunofluorescence (IF) assay to detect the expression of tyrosine hydroxylase (TH) neurons，content of TH-positive fibers in the striatum and to detect the activation status of nigrostriatal microglia in the mouse midbrain; Real-time PCR was employed to measure the levels of interleukin (IL)-6, IL-1β, and tumor necrosis factor-α (TNF-α) in the substantia nigra of the mouse brain. Western blotting was used to measure the protein levels of TH, nuclear factor-κB (NF-κB) p65, NF-κB inhibitor α (IκBα), and phosphorylated IκBα (p-IκBα) in the substantia nigra of the mouse brain.  Results  Compared to the control group and the solvent control group, the rotenone-induced PD model group exhibited motor impairments in behavioral tests, a decrease in the number of TH positive neurons in the substantia nigra (P <0.0001), decreased levels of TH-positive fibers in the striatum, activation of midbrain substantia microglia，and elevated levels of IL-6,  IL-1β, TNF-α, p-IκBα, and NF-κB p65 expression. ACT001 significantly improved the behavioral impairments and substantia nigra damage in PD mice, increased the number of TH-positive neurons in the substantia nigra, increased levels of TH-positive fibers in the striatum, inhibition of microglial cell activation in the midbrain substantia nigra, and elevated the protein expression levels of IκBα while reducing the levels of IL-6, IL-1β, TNFα, -IκBα, and NF-κB p65 in the substantia nigra ( P<0.05). At a dose of 5 mg/kg, ACT001 significantly improved behavioral impairments in rotenone-induced PD mice, reduced the loss of dopaminergic neurons, and its mechanism may be related to the inhibition of the NF-κB  signaling pathway and the suppression of inflammation. In summary, the intervention of ACT001 in the rotenone-induced PD mouse model inhibited the inflammatory response in the midbrain, increased the number of TH-positive neurons, and augmented the population of dopaminergic neurons in the substantia nigra, exerting a protective effect on neurons.   Conclusion  ACT001 significantly improves behavioral deficits in ROT-induced PD mice, ameliorates of dopaminergic neuron loss from the midbrain substantia nigra and striatum, inhibits the activation of nigrostriatal microglia in the midbrain, and suppresses inflammatory responses by inhibiting the activation of the NF-κB signaling pathway. 

Objective To explore the pathogenesis of Alzheimer’s disease by examining the effects of dihydroartemisinin(DHA) on cognitive behavior, hippocampal, cerebral cortex and retinal cell morphology, β-amyloid(Aβ) and autophagy-related proteins in 5×FAD mice.   Methods  Twenty 5×FAD mice and 5 wild type (WT) mice were selected, all of which were female. The 5×FAD mice were randomly divided into model (M) group, donepezil (D) group, low-dose DHA (DHA-L) group, and high-dose DHA (DHA-H) group. The WT and M groups were not treated, and the D group was given donepezil 0.1 mg/kg per day. DHA-L group and DHA-H group were given 10 mg/kg and 20 mg/kg DHA per day, respectively. Group D, group DHA-L and group DHA-H were given intragastric administration once a day for 3 months. The changes of in cognitive behavior were measured by Morris experiment. HE staining was used to observe the arrangement and morphology of nerve cells in cerebral cortex, hippocampus and retina. The expressions of Aβ protein in cerebral cortex, hippocampus and retina were detected by immunohistochemistry. Western blotting detected the expression of autophagy related proteins (LC3-Ⅰ, LC3-Ⅱ, Beclin-1, P62, β-actin).   Results  The DHA-H group and the D group exhibited more frequent adoption of both linear and trending exploration routes. Compared to the model group, significant differences in the contents of Aβ  in the hippocampal CA1, cerebral cortex S1, and retinal were observed (P<0.0001) in the other four groups. The analysis also showed significant differences in autophagy-associated proteins between the DHA-L,  DHA-H, and model groups (P<0.01).  Conclusion  DHA improves cognitive function and increases the number of nerve cells in mice. It also reduces Aβ content in the cerebral cortex, hippocampus, and retina, along with improving autophagy-associated protein deposition in mice.  

Objective  To investigate the effect of up-regulating tumor necrosis factor alpha induced protein 3 (TNFAIP3) expression on hippocampal neurons in mice with cerebral ischemia-reperfusion.    Methods  The mice were randomly divided into 6 groups: sham group, sham empty vector group (sham-), sham TNFAIP3 high expression group (sham+), model group, model empty vector group (model- ), model TNFAIP3 high expression group (model+ ). A mouse model of middle cerebral artery occlusion and cerebral ischemia-reperfusion was established using the suture method. After the successful establishment of the model, lentivirus was injected into the hippocampus 24 hours later. Two weeks later, samples were collected and Western blotting was used to detect the expressions of TNFAIP3 and ERK signaling pathway proteins. The changes in ischemic area were observed by 2,3,5-triphenyltetrazolium chloride (TTC) staining; HE staining was used to observe the morphological changes of hippocampal neurons, and ELISA was used to detect the expressions of lipoprotein-associated phospholipase A2 (Lp-PLA2) and interleukin (IL)-8.     Results  The results of Western blotting indicated that the TNFAIP3 expression in the model group decreased significantly compared with the sham group (P<0.05); Compared with the model group, there was no significant change in TNFAIP3 expression in the model-  group (P>0.05); The TNFAIP3 expression in the model+  group increased significantly compared with the model group and model-  group(P<0.05). Compared with the sham group, the results of the sham+  group showed that the ischemic area had no significant changes in TTC staining, and there were no significant changes in hippocampal neuronal morphology, and the expressions ERK signaling pathway proteins, Lp-PLA2 and IL-8 (P>0.05); Compared with the sham-  and sham+  groups, the model group showed an increase in ischemic area, significant damage to hippocampal neurons, a decrease in the number of Nissl bodies, and a significant increase in the expressions of ERK signaling pathway proteins, Lp-PLA2, and IL-8 (P<0.05); Compared to the model-  group, the model+  group showed a decrease in ischemic area, an increase in the number of neurons in the hippocampus and the number of Nissl bodies, and a significant decrease in the expressions of ERK signaling pathway proteins, Lp-PLA2, and I-8 (P<0.05).   Conclusion  Up-regulation of TNFAIP3 may be one of the methods for repairing hippocampal neuronal damage caused by cerebral ischemia reperfusion.

 Objective To evaluate the changes in postoperative plasma β-endorphin(β-EP) levels in patients who had received perioperative electroacupuncture（EA） treatment in 10 randomized controlled trials (RCTs) and examine the impact of EA on postoperative pain.   Methods  This meta-analysis evaluated the changes in plasma β-EP levels and visual analog scale (VAS) 12, 24 and 48 hours after surgery in patients receiving perioperative EA. It also assessed the changes in plasma serotonin (5-hydroxytryptamine, 5-HT) and prostaglandin E2 (PGE2) levels at 24 hours postsurgery. A comprehensive search was conducted in the China National Knowledge Infrastructure (CNKI), Wanfang, Chongqing VIP database, Chinese Biomedical Database (CBM), Web of Science, and PubMed databases. RCTs on perioperative EA and β-EP published from the inception of the websites up to July 25, 2023, were retrieved. Effect size aggregation, literature quality assessment, and bias analysis were performed using RevMan 5.3 software, and sensitivity analysis was conducted via R 4.3.1.   Results  A total of 10 RCTs involving 706 patients were included. EA in conjunction with conventional anesthesia significantly increased plasma β-EP levels at 12 hours postsurgery ［standard mean difference (SMD)=2.79, 95% CI (1.85, 3.72), Z=5.81, P<0.00001］, 24 hours postsurgery ［SMD=1.87, 95% CI (0.9, 2.83), Z=3.79, P=0.0001］, and 48 hours postsurgery ［SMD=2.02, 95% CI (1.49, 2.54), Z=7.50, P<0.00001］. EA reduced plasma PGE2 levels at 24 hours postsurgery and plasma 5-HT levels at 24 hours postsurgery, and the VAS at 12, 24 and 48 hours after surgery also decreased.   Conclusion  These findings suggest that perioperative EA markedly elevates plasma β-EP levels, reduces pain-inducing factors in plasma, and effectively alleviates acute postoperative pain.

Objective To investigate the expressions of annexin A2 and glycogen synthesis kinase-3β (GSK-3β) in cutaneous squamous cell carcinoma (CSCC) tissues, and to analyze their correlation with CSCC as well as their clinical pathological diagnostic value.   Methods  The pathological tissues of 68 patients with CSCC and 40 patients with keratoacanthoma (KA) who underwent surgical treatment in the Department of Dermatology of the Second Hospital of Nanning from October 2020 to May 2024 were collected, and the surrounding normal skin tissues of 32 patients with benign skin diseases were used as controls. The expressions of annexin A2, GSK-3β and β-catenin were detected by immunohistochemistry and Western blotting. Spearman was used to evaluate the correlation between the expressions of annexin A2 and GSK-3β and the pathological characteristics in CSCC. The receiver operating characteristic (ROC) curve was drawn to analyze the clinical diagnostic value of annexin A2 and GSK-3β in CSCC.  Results Compared with the normal skin tissues, the expressions of annexin A2 and βcatenin in CSCC increased, and GSK-3β  decreased (P<0.05); Compared with the KA tissues, the expression of annexin A2 in CSCC tissues increased (P<0.05). The expression of annexin A2 was negatively correlated with that of GSK-3β in CSCC (r=-0.3901, P<0.01). GSK-3β expression was related to tissue differentiation, with lower expression in poorly differentiated patients’ cancer tissues (P<0.05). The sensitivity of annexin A2 and GSK-3β for diagnosis of CSCC was 85.3% and 41.2%, respectively, with specificities of 46.9% and 84.4% respectively. The sensitivity of annexin A2 for distinguishing between CSCC and KA was 85.3%, with a specificity of 40.0%.   Conclusion  Annexin A2 and GSK-3β may be used as potential biomarkers for the early diagnosis or differential diagnosis of CSCC, and play important roles in the development of CSCC. Their mechanism may be related to the activation of Wnt/β-catenin signaling pathway. 

Objective  To explore the anatomical landmarks and segmentation method  for the intraoperative identification of the cervical segment of the internal carotid artery by studying cadaveric dissections with an endoscopic transoral medial pterygomandibular fold approach and to investigate its clinical significance.   Methods  The head specimens of five fresh frozen cadavers were dissected in the anatomical laboratory of the Surgical Treatment Technology Innovation Unit of Nasal Skull Base Tumor in Eye & ENT Hospital of Fudan University. The parapharyngeal space was dissected layer by layer through the endoscopic transoral medial pterygomandibular fold approach, and the location marks of parapharyngeal internal carotid artery (ppICA) and adjacent structures of ppICA were anatomically studied. The anatomical landmarks associated with ppICA were observed and characterized, and the ppICA was segmented anatomically according to its adjacent structures. Then, the length of each ppICA segment was measured.  Results  Muscle structures were essential anatomical landmarks for an endoscopic transoral pterygoid medial approach that identifies mandibular folds. The first layer of muscles included the superior pharyngeal constrictor, tensor veli palatini, and medial pterygoid muscles. The second layer includes the stylopharyngeus, styloglossus, longus capitis, and levator veli palatini muscles. The stylopharyngeal and levator veli palatini muscles were close to the ppICA and were reliable landmarks for locating the ppICA. Furthermore, the ppICA was divided into three segments according to their positional relationship with the ppICA. The first segment of ppICA(P1ICA) was located between the greater horn plane of the hyoid bone and the intersection plane between the upper margin of stylopharyngeal muscle and ppICA. The second segment of ppICA (P2ICA) was between the plane where the upper edge of the stylopharyngeal muscle intersected with the ppICA and the plane where the projection of inferior edge of the levator veli palatini muscle intersected with the ppICA. The third segment of ppICA (P3ICA) was between the intersection of the lower margin projection of the levator veli palatini muscle and ppICA and the external orifice of the carotid canal. The P2ICA was within an anatomical region bounded by the levator veli palatini muscle, longus capitis muscle, and stylopharyngeus muscle. This region was termed “ICA window” in this paper measured under the cadaver head specimen, the lengths of P1ICA, P2ICA, and P3ICA were (36.5±7.3) mm, (15.5±1.6) mm, (7.4±1.7) mm respectively.   Conclusion  The muscular structure refers to the relatively constant anatomical reference landmarks within the endoscopic transoral medial pterygomandibular fold. The stylopharyngeus and levator veli palatini muscles are reliable landmarks for precisely locating and segmenting the ppICA, thus having essential clinical implications.  

Objective To investigate the anatomical and microscopic structures of interstitial fluid flow channels in the skin tissue of hand dorsum in human cadavers.  Methods  Totally 7 fresh cadavers within 12 hours post-mortem were included. MRI was used to observe the distribution of interstitial fluid flow from the first phalanx of the fingers to the wrist, precisely locating the flow channels. Based on imaging results, histological analyses were conducted to determine the histological characteristics of the flow channels. Furthermore, multi-immunofluorescence and microcomputed tomography (Micro-CT) techniques were employed to analyze the channels, and image post-processing was used to elucidate their anatomical structures at the microscopic level.   Results  After injecting a contrast agent into the first phalanx of ten finger specimens and applying repeated pressure, MRI image revealed centripetal long-range interstitial fluid flow along channels distinct from blood vessels and lymphatic vessels. Histological analysis and Micro-CT further confirmed that the flow primarily occurred within the fibrous connective tissue and adventitia of the skin.   Conclusion  The orderly fibrous connective tissue and adventitia in the skin form the interstitial fluid flow channels in the human hand dorsum skin, named as “stromal membrane channels” in the skin. 

Objective  To construct a finite element analysis model for the treatment of calcaneal fracture for Sanders Ⅲ type calcaneal by using simulated sustentaculum tali sustentacular screw placement, and to explore the effectiveness of the treatment of Sanders type Ⅲ calcaneal fracture.    Methods  The finite element analysis method  was used to study the effectiveness of the treatment of Sanders Ⅲ calcaneal fracture. Three calcaneal specimens of a healthy adult were taken for Micro-CT scanning to obtain the CT sections data, then Mimics 21.0 and Geomagic Wrap 2017 software were used to reconstruct the three-dimensional model of normal calcaneal bone, and SolidWorks 2017 software was used to map the internal fixation and fracture model according to the clinical fracture cases, and simulated surgery was performed. The data obtained were imported into ANSYS 17.0 finite element analysis software for material assignment and mesh division to establish a three-dimensional finite element model. The load and boundary constraints were applied to each model to perform finite element analysis and calculation, and then the stress of the finite element model and the maximum displacement of the fracture end were extracted.  Results  The maximum stress of the overall structure of each model was concentrated on internal fixation, in which the maximum stress of sustentaculum tali fracture and the maximum stress of screws in all fracture models are located on the sustentaculum tali sustentacular screw. The displacement of fracture end in each finite element model was less than 0.15 mm, and no internal fixation failure occurred.  Conclusion  It is effective to simulate the treatment of calcaneal fracture of Sanders type Ⅲ by using of sustentaculum tali sustentacular screw.  

 Objective  To explore the relationship between NANOS3 and P-bodies in oocytes and the mechanism of their interaction during early oogenesis.  Methods  The co-localization of NANOS3 and dead box helicase 6(DDX6) in day post postnatal 1 (1dpp) mouse oocytes was observed by immunofluorescence, and the interaction between NANOS3 and DDX6 was detected by immunoprecipitation. NANOS3 and DDX6 full-length plasmids were constructed to transfect HEK293T cells, and the mechanism of their interaction was investigated by immunofluorescence and immunoprecipitation. NANOS3 transfected HeLa cells to investigate whether NANOS3 had the ability of liquid-liquid phase separation (LLPS) by live-cell imaging. The proteins recruited by P-bodies in early oogenesis were identified by DDX6-immunoprecipitation-mass spectrometry(DDX6-IP-MS).    Results  NANOS3 and DDX6 colocalized and interacted with each other in 1dpp mouse oocytes. However, the co-localization of NANOS3 and DDX6 was not observed in HEK293T cells that had been transfected, but co-immunoprecipitation still demonstrated an interaction between these two proteins. Besides, live-cell imaging revealed that NANOS3 exhibited dynamic fluid-like properties within cells, which may promote the formation of P-bodies through LLPS. Finally, DDX6-IP-MS revealed that DDX6 might recruit NANOS3 into P-bodies by binding to the NANOS3 interacting protein Pumilio.   Conclusion  NANOS3 serves as a specific component of P-bodies in neonatal oocytes and may be involved in the regulation of early oogenesis. 

Objective To investigate gender differences and age-related changes in body composition(BC) among Miao adults in Rongshui, Guangxi Province, and to provide the basis for assessing nutritional status and health.   Methods  With informed consent, 630 Miao adults (218 males, 412 females) were randomly selected for this study. Body composition was assessed using bioelectrical impedance analysis(BIA).   Results  Weight, fat-free mass, muscle mass, trunk muscle mass, limb muscle mass, waist-to-hip ratio(WHR), body water, presumtion of bone mass and protein were significantly higher in males than in females. And the fat mass, trunk fat mass, limb fat mass, visceral fat content, subcutaneous fat content and percentage of body fat were significantly higher in females than in males. According to the evaluation of body mass index(BMI) and WHR, the proportion of overweight and obesity of Miao adults was higher than the average level of Miao residents, and their obesity was characterized by central obesity. With age, weight, fat mass, muscle mass, fat-free mass, limb muscle mass, limb fat mass, subcutaneous fat content, percentage of body fat, body water, presumtion of bone mass, and protein of Rongshui Miao adults showed a gradual decreasing trend, while visceral fat content and WHR increased progressively. BMI in male Miao adults, along with BMI, fat mass, trunk fat mass, subcutaneous fat content, percentage of body fat, and body water in female Miao adults, showed a trend of increasing followed by decreasing, peaking at the age of 40-49  years.    Conclusion  The body composition of Miao adults in Rongshui, Guangxi, exhibits significant gender differences and age-related variation change patterns, which may increase the risk of sarcopenia and metabolic diseases with aging.  

 Objective  To analyze the relationship between obesity indicators，muscle mass indices and hypertension in Dongxiang adults, and establish and evaluate a Nomogram model based on these indicators used to predict the risk of hypertension in this population.    Methods  A total of 1209 Dongxiang adults from Linxia Prefecture, Gansu Province were selected, 11 obesity indicators and 5 muscle mass indicators，including neck circumference (NC), waist-to-hip ratio (WHR), BMI, ponderal index (PI), conicity index (CI), a body shape index (ABSI), body roundness index (BRI), abdominal volume index (AVI), hip index (HI), body adiposity index (BAI), and the ratio of limb fat mass to body weight(LFWR), appendicular skeletal muscle mass (ASM), appendicular skeletal muscle mass index (ASMI), appendicular skeletal muscle mass to BMI ratio (ASMBMI), skeletal muscle index (SMI), and trunk muscle mass to body weight ratio(TMWR) were measured. Logistic regression analysis was used to explore the relationship between each indicator and hypertension. Nomogram prediction model was constructed and validated by using R language.   Results Among Dongxiang males, the NC, BMI, PI, WHR, CI, AVI, BRI, BAI, ASM, ASMI, and LFWR were lower in the normal and highnormal blood pressure groups compared to the hypertensive group, while SMI, ASMBMI, and TMWR were higher. Among females, similar trends were observed, with lower NC, BMI, PI, WHR, CI, AVI, BRI, BAI, and LFWR in the normal blood pressure and high-normal blood pressure groups compared to the hypertensive group, but SMI, ASMBMI, and TMWR higher than the hypertensive group (P<0.05). Results of multivariate Logistic regression analysis indicated that age, NC, and WHR were risk factors for hypertension in Dongxiang adults, while was protective factor (P<0.05). The area under curve(AUC) of the Nomogram model constructed based on these factors was 0.796, and the Bootstrap internal validation C-index was 0.7957, indicating good calibration of the model.    Conclusion  The Nomogram model constructed based on obesity and muscle mass indicators has good predictive efficiency for predicting the risk of hypertension in Dongxiang adults. 

Objective  To investigate the effect of Gd-hydroxyapatite(Gd-HA) stents with adipose mesenchymal cells (ADSCs) on the repair of knee articular cartilage defects.     Methods  To isolate, culture, and identify rabbit ADSCs by establishing a rabbit knee joint full-thickness cartilage defect model, a total of 18 rabbits were randomly divided into blank control group, Gd-HA scaffold group, and ADSCs+Gd-HA scaffold group. At week 12 and 24 after surgery, International Curtilage Repair Society(ICRS) score, HE, toluidine blue, modified red O bright green and ColⅡ were detected by immunohistochemical staining, then ColⅡ and GAG mRNA expression levels were detected by O’Driscoll and Realtime PCR. ColⅡ protein expression was detected by Western blotting, GAG content was detected by DMMB, biomechanical strength was detected by indentation test, and PKH26 labeled ADSCs was used to trace the tissue engineering scaffold with Gd-HA composite ADSCs to evaluate the repair effect of rabbit knee cartilage defects.    Results  The ADSCs isolated and cultured in vitro  showed good growth, stable phenotype and good directional differentiation through macroscopic observation and histological staining, it could be seen that the repair degree and effect of the knee joint fullthickness cartilage defect model implanted with Gd-HA scaffold group were better than those of the blank control group, while the cartilage repair situation of the ADSCs+Gd-HA scaffold group  was better than that of the Gd-HA scaffold group (P<0.05);  The ICRS and improved O’Driscoll scores were higher than the other two groups (P<0.05). Compared with the Gd-HA group, the ADSCs+Gd-HA group could produce ColⅡ and GAG during the process of cartilage repair, with stronger mechanical strength of the repaired tissue (P<0.05); PKH26 labeled ADSCs were found in the repaired tissues of the ADSCs+Gd-HA group, and they were involved in the composition of newly formed tissues.   Conclusion  Gd-HA scaffold material combined with ADSCs has a good repair effect on full-thickness cartilage defects in the knee joint as a new type of biological material for repairing joint cartilage defects. 

Objective  To explore an experimental protocol for differentiating humaninduced pluripotent stem cells (iPSCs) into highly pure midbrain dopaminergic (DA) neurons.    Methods  By optimizing a blend of small molecules and recombinant human growth factors, iPSCs were induced to differentiate into ventral midbrain floor plate DA progenitor cells and subsequently into mature substantia nigra pars compacta DA neurons. Throughout the differentiation process, Real-time PCR and immunofluorescent staining were utilized as a method  for quality assessment. Results  iPSCs firstly differentiate into dopaminergic precursor cells, and then gradually differentiate into DA neurons expressing tyrosine hydroxylase (TH).     Conclusion  The protocol successfully yields approximately high purity tyrosine hydroxylase-positive (TH+) DA neurons. This differentiation technique offers an effective cellular model for studying the physiological mechanisms and pathogenesis of Parkinson’s disease, providing valuable insights for future research and potential therapeutic strategies. 

 In recent years, more and more researches has focused on the correlation between cognitive activity and physiological variables. The change of pupil is regarded as an important target in the cognitive process, and has become a hot research field. This review focuses on the three key brain regions that regulate pupil change, and reflects the neurophysiological mechanism behind pupil change by elaborating the neural pathways related to pupil change and cognitive performance. Based on recent studies on pupil change in cognitive diseases, it aims to promote the application of pupil change in the field of cognitive science in the future. 

With the rapid development of the Internet, the problem of internet gaming disorder (IGD) among adolescents has gradually become prominent. IGD has caused serious harm to their physical and mental health, and it is difficult to withdraw and treat. In 2019, the World Health Organization (WHO) officially listed Gaming Disorder (GD) in the diagnostic category and included it in the 11th edition of the International Classification of Diseases. In China, the number of adolescent internet users has approached 200 million, and the number of IGD patients is relatively large. It is urgent to make breakthroughs in the cognition and treatment of IGD. Domestic and foreign studies have shown that IGD’s physical and psychological harm to adolescents is related to the abnormal activation and functional connection damage of structures such as the prefrontal lobe, limbic system, and striatum, which in turn causes neurocognitive disorders, executive dysfunction, difficulty in emotional regulation, abnormal reward and punishment feedback, behavioral abnormalities and other functional disorders; therefore, this article aims to systematically review the research literature on IGD in recent years, and use neurophysiological imaging research method  to sort out the changes in the brain structure and function of adolescents with IGD, in order to improve the understanding of the pathophysiology of the disease, and assist in further clinical diagnosis, treatment, application and research design. 

 Tumor is the main lethal type of major diseases that human beings are facing nowadays. Traditional therapies, such as chemotherapy, surgery, radiotherapy, etc, have been the mainstay of tumor treatment for a long time, and still have their own limitations in terms of effectiveness. In recent years, immunotherapy for tumors has attracted much attention with the rise of strategies such as immune checkpoint inhibitors, chimeric antigen receptor T-cell (CAR-T) relay therapy, and anti-tumor vaccines. Among them, CAR-T cell therapy, as a breakthrough tumor immunotherapy method, can precisely target tumors at the cellular and molecular levels by modifying the patient’s own T cells to express recombinant antigen receptors targeting specific tumor antigens, breaking the restriction of the major histocompatibility complex (MHC) of the natural T cells themselves. It can precisely target tumors at the cellular and molecular levels, and thus can efficiently and specifically identify and kill cancer cells, providing a new perspective for treatment. Since the approval of the first CAR-T therapy by the US Food and Drug Administration (FDA) in 2017, in addition to still maintaining significant advantages in the field of classical multiple hematologic malignancies, it has also made outstanding progress in the treatment and clinical trials of some solid tumors and autoimmune diseases, among others. This paper  will briefly review the current principles, applications, challenges and directions of development of CAR-T technology.