N7-methylguanosine-related miRNAs predict hepatocellular carcinoma prognosis and immune therapy.

N7-methylguanosine (m7G) modification has been notably linked with the development of many tumors. However, no investigations have been conducted on whether m7G-related miRNA (m7G-miRNA) is a prognostic index of hepatocellular carcinoma (HCC). Therefore, this investigation aimed to establish a predictive m7G-miRNA signature for efficient HCC prognosis and elucidate the associated immune cell infiltration (ICI) and functions in the tumor microenvironment. RNA sequencing and clinical data on 375 HCC and 50 healthy tissue samples were acquired from The Cancer Genome Atlas database. The m7G-miRNA regulators methyltransferase-like 1 and WD repeat domain 4 were acquired from the TargetScan database. Univariate Cox regression analysis was conducted on the 63 differentially expressed m7G-miRNAs identified. A prognostic signature that consisted of seven miRNAs was identified. According to their risk scores, individuals with HCC were divided into high-risk (HR) and low-risk (LR) cohorts. A Kaplan-Meier test revealed that survival in the HR HCC patients was poorer than in the LR cohort (p < 0.001). The area under the receiver operating characteristic curves of 1-, 3-, and 5-year overall survival were 0.706, 0.695, and 0.715, respectively. A nomogram of sex, risk score, age, and stage indicated the HCC patients' overall survival. Furthermore, it was indicated that the HR and LR patients had different degrees of ICI and immune function. A pathway enrichment analysis revealed the association of several immunity-linked pathways with the risk model. In conclusion, the signature established has great prognostic value and could be used as a new immunotherapy target for individuals with HCC.

Characteristics of spontaneous nystagmus and its correlation to video head impulse test findings in vestibular neuritis.

Objective: To explore the direction and SPV (slow phase velocity) of the components of spontaneous nystagmus (SN) in patients with vestibular neuritis (VN) and the correlation between SN components and affected semicircular canals (SCCs). Additionally, we aimed to elucidate the role of directional features of peripheral SN in diagnosing acute vestibular syndrome. Materials and methods: A retrospective analysis was conducted on 38 patients diagnosed with VN in our hospital between 2022 and 2023. The direction and SPV of SN components recorded with three-dimensional videonystagmography (3D-VNG) and the video head impulse test (vHIT) gain of each SCC were analyzed as observational indicators. We examined the correlation between superior and inferior vestibular nerve damage and the direction and SPV of SN components, and vHIT gain values in VN patients. Results: The median illness duration of between symptom onset and moment of testing was 6 days among the 38 VN patients (17 right VN and 21 left VN). In total, 31 patients had superior vestibular neuritis (SVN), and 7 had total vestibular neuritis (TVN). Among the 38 VN patients, all had horizontal component with an SPV of (7.66 ± 5.37) °/s, 25 (65.8%) had vertical upward component with a SPV of (2.64 ± 1.63) °/s, and 26 (68.4%) had torsional component with a SPV of (4.40 ± 3.12) °/s. The vHIT results in the 38 VN patients showed that the angular vestibulo-ocular reflex (aVOR) gain of the anterior (A), lateral (L), and posterior (P) SCCs on the ipsilesional side were 0.60 ± 0.23, 0.44 ± 0.15 and 0.89 ± 0.19, respectively, while the gains on the opposite side were 0.95 ± 0.14, 0.91 ± 0.08, and 0.96 ± 0.11, respectively. There was a statistically significant difference in the aVOR gain between the A-, L-SCC on the ipsilesional side and the other SCCs (p < 0.001). The aVOR gains of A-, L-, and P-SCC on the ipsilesional sides in 31 SVN patients were 0.62 ± 0.24, 0.45 ± 0.16, and 0.96 ± 0.10, while the aVOR gains on the opposite side were 0.96 ± 0.13, 0.91 ± 0.06, and 0.98 ± 0.11, respectively. There was a statistically significant difference in the aVOR gain between the A-, L-SCC on the ipsilesional side and the other SCCs (p < 0.001). In 7 TVN patients, the aVOR gains of A-, L-, and P-SCC on the ipsilesional side were 0.50 ± 0.14, 0.38 ± 0.06, and 0.53 ± 0.07, while the aVOR gains on the opposite side were 0.93 ± 0.17, 0.90 ± 0.16, and 0.89 ± 0.09, respectively. There was a statistically significant difference in the aVOR gain between the A-, L-, and P-SCC on the ipsilesional side and the other SCCs (p < 0.001). The aVOR gain asymmetry of L-SCCs in 38 VN was 36.3%. The aVOR gain asymmetry between bilateral A-SCCs and bilateral P-SCCs for VN patients with and without a vertical upward component was 12.8% and 8.3%, which was statistically significant (p < 0.05). For VN patients with and without a torsional component, the aVOR gain asymmetry of bilateral vertical SCCs was 17.0% and 6.6%, which was statistically significant (p < 0.01). Further analysis revealed a significant positive correlation between the aVOR gain asymmetry of L-SCCs and the SPV of the horizontal component of SN in all VN patients (r = 0.484, p < 0.01), as well as between the asymmetry of bilateral vertical SCCs and the SPV of torsional component in 26 VN patients (r = 0.445, p < 0.05). However, there was no significant correlation between the aVOR gains asymmetry of bilateral A-SCCs and P-SCCs and the SPV of the vertical component in 25 VN patients. Conclusion: There is a correlation between the three-dimensional direction and SPV characteristics of SN and the aVOR gain of vHIT in VN patients. These direction characteristics can help assess different SCCs impairments in patients with unilateral vestibular diseases.

[Further exploration of the classification and clinical value of head-shaking nystagmus].

Objective:To investigate the classification of head shaking nystagmus（HSN） and its clinical value in vestibular peripheral diseases. Methods:Clinical data of 198 patients with peripheral vestibular disorders presenting with HSN were retrospectively analyzed. Video Nystagmograph（VNG） was applied to detect spontaneous nystagmus（SN）, HSN, and Caloric Test（CT）. The intensity and direction of SN and HSN as well as the unilateral weakness（UW） and direction preponderance（DP） values in caloric test was analyzed in patients. Results:Among the 198 patients with vestibular peripheral disease, there were 105 males and 93 females, with an average age of（49.1±14.4） years （range: 14-87 years）. One hundred and thirty seven patients were diagnosed as Vestibular Neuritis（VN）, 12 as Meniere's Disease（MD）, 41 as sudden deafness（SD） and 8 as Hunt's syndrome accompanied by vertigo. Among them, there were 116 patients in the acute phase, including 68 cases（58.6%） with decreased HSN, 4 cases（3.4%） with increased HSN, 5 cases（4.3%） with biphasic HSN, 38 cases（32.8%） with unchanged HSN, and 1 case（0.9%） with perverted HSN. There were 82 cases in the non-acute phase, 51 cases（62.2%） with decreased HSN, 3 cases（3.6%） with increased HSN, 9 cases（11.0%） with biphasic HSN, and 19 cases（23.2%） with unchanged HSN. In biphasic HSN, the intensity of phase I nystagmus was usually greater than that of phase II, and the difference was statistically significant（P<0.01）. There was no correlation between HSN type and course of disease or DP value. The intensity of HSN was negatively correlated with the course of disease（r=-0.320, P<0.001） and positively correlated with DP value（r=0.364, P<0.001）, respectively. The intensity of unchanged nystagmus and spontaneous nystagmus were（8.0±5.7） °/s and（8.5±6.4）°/s, respectively. There was no statistically significant difference in the intensity of nystagmus before and after shaking the head. Conclusion:HSN can be classified into five types and could be regarded as a potential SN within a specific frequency range （mid-frequency）. Similarly, SN could also be considered as a common sign of unilateral vestibular impairment at different frequencies. HSN intensity can reflect the dynamic process of vestibular compensation, and is valuable for assessing the frequency of damage in peripheral vestibular diseases and monitoring the progress of vestibular rehabilitation.

Three-dimensional characteristics of nystagmus induced by low frequency in semicircular canals of healthy young people.

Objective: The study aimed to analyze the three-dimensional characteristics of nystagmus induced by different semicircular canal combinations in healthy young people, and to determine the reference range of nystagmus slow phase velocity (SPV) and its asymmetry. Materials and methods: Fifty-two healthy volunteers (26 males and 26 females, aged 17-42 years, average 23.52 ± 6.59), were recruited to perform the manual triaxial rotation testing with a 3D-Videonystagmography (3D-VNG) device (VertiGoggles (ZT-VNG-II), Shanghai ZEHNIT Medical Technology Co., Ltd., Shanghai, China) using a 0.3 Hz prompt beat and a 90° amplitude, respectively. The induced nystagmus around the Z-, X-, and Y-axes were recorded in the yaw, pitch, and roll planes. The directions and slow phase velocities of the horizontal, vertical, and torsional components of the induced nystagmus under different semicircular canal combinations (the left lateral and right lateral semicircular canal combination, bilateral anterior semicircular canals, bilateral posterior semicircular canals combination, and the anterior and posterior semicircular canals combination of each ear), as well as their asymmetry, were taken as the observation indexes to analyze the characteristics of the nystagmus vectors of different combinations. Results: Fifty-two healthy volunteers had no spontaneous nystagmus. The characteristic nystagmus was induced by the same head movement direction in all three axial rotation tests. The SPVs of the left and right nystagmus were 44.45 ± 15.75°/s and 43.79 ± 5.42°/s, respectively, when the subjects' heads were turned left or right around the Z-axis (yaw). The SPVs of vertically upward and downward nystagmus were 31.67 ± 9.46°/s and 30.01 ± 9.20°/s, respectively, when the subjects' heads were pitched around the X-axis (pitch). The SPVs of torsional nystagmus, with the upper poles of the eyes twisting slowly to the right and left ears (from the participant's perspective), were 28.99 ± 9.20°/s and 28.35 ± 8.17°/s, respectively, when the subjects' heads were turned left or right around the Y-axis (roll). There was no significant difference in the SPVs of nystagmus induced by the same rotation axis in two opposite directions (p > 0.05). The reference ranges for the slow phase velocities (SPVs) of nystagmus induced by the triaxial rotation testing were as follows: For the Z-axis (yaw), the SPVs were 13.58-75.32°/s for leftward head rotation and 13.56-74.02°/s for rightward head rotation. For the X-axis (pitch), the SPVs were 13.13-50.21°/s for upward head nystagmus and 11.98-48.04°/s for downward head nystagmus. For the Y-axis (roll), the SPVs were 10.97-47.02°/s for the left-sided head rotation and 12.34-44.35°/s for the right-sided head rotation. Conclusion: This study clarified the three-dimensional characteristics of nystagmus induced by different semicircular canal combinations in healthy young people. It also established a preliminary reference range of SPVs and SPV asymmetry of nystagmus induced by the vertical semicircular canal. It can further provide a basis for the mechanism of semicircular canal-induced nystagmus and the traceability of nystagmus in patients with otogenic vertigo. It is shown that the portable 3D-VNG eye mask can be used for the manual triaxial rotation testing to achieve the evaluation of the low-frequency angular vestibulo-ocular reflex (aVOR) function of the vertical semicircular canal, which is convenient, efficient, and practical.

The 3D characteristics of nystagmus in posterior semicircular canal benign paroxysmal positional vertigo.

Objective: The aim of this study was to observe the 3-dimensional (3D; horizontal, vertical, and torsional) characteristics of nystagmus in patients with posterior semicircular canal canalithiasis (PSC-can)-related benign paroxysmal positional vertigo (BPPV) and investigate its correlation with Ewald's. Methods: In all, 84 patients with PSC-can were enrolled. The latency, duration, direction, and slow-phase velocity induced by the Dix-Hallpike test in the head-hanging and sitting positions were recorded using 3D video nystagmography (3D-VNG). The characteristics of the horizontal, vertical, and torsional components of nystagmus were quantitatively analyzed. Results: 3D-VNG showed that the fast phase of the vertical components and torsional components of left and right ear PSC-can as induced by the head-hanging position of the Dix-Hallpike test were upward, clockwise and counterclockwise, and horizontal components were mainly contralateral. The median slow-phase velocity of each of the three components for consecutive 5 s was 26.3°/s (12.3-45.8), 25.0°/s (15.7-38.9), and 9.2°/s (4.9-13.7). When patients were returned to the sitting position, the fast phase of the vertical and torsional components of nystagmus was reversed. Only 54 patients had horizontal components of nystagmus, and 32 of them remained in the same direction. The median slow-phase velocity of the three components for consecutive 5 s was 9.4°/s (6.0-11.7), 6.8°/s (4.5-11.8), and 4.9°/s (2.8-8.0). The ratios of the slow-phase velocity of the horizontal, vertical, and torsional components of the head-hanging position to the sitting position were close to 1.85 (1.0-6.6), 3.7 (1.9-6.6), and 5.1 (2.6-11.3). The ratios of the slow-phase velocity of the vertical to horizontal component, the torsional to horizontal component, and the vertical to torsional component of the head-hanging position were close to 3.3 (1.7-7.6), 3.9 (1.8-7.6), and 1.0 (0.5-1.8). The ratios of the slow-phase velocity of the vertical to horizontal component, the torsional to horizontal component, and the vertical to torsional component of the sitting position were close to 2.1 (1.1-6.8), 1.5 (1.0-3.8), and 1.2 (0.8-2.8). Conclusion: There were three components of nystagmus induced by the Dix-Hallpike test in patients with PSC-can. The vertical component was the strongest and the horizontal component was the weakest. The 3D characteristics of nystagmus were consistent with those of physiological nystagmus associated with the same PSC with a single-factor stimulus, in accordance with Ewald's law.

The horizontal and vertical components of nystagmus evoked by the supine roll test in horizontal semicircular canal canalolithiasis.

Objective: The characteristics of horizontal and vertical components of nystagmus evoked by the supine roll test in patients with horizontal semicircular canal canalolithiasis (HSC-Can) were analyzed, according to Ewald's first law. It provided a basis for the study of human horizontal semicircular canal function and structure, objective diagnosis, and treatment of benign paroxysmal positional vertigo (BPPV). Materials and methods: The records of patients that had been tested with 2-dimensional videonystagmography (2D-VNG) were reviewed between June 2019 and June 2021. The intensity and direction of horizontal and vertical nystagmus elicited by the supine roll test were analyzed in 189 patients with idiopathic HSC-Can. Results: All the 189 patients with HSC-Can were induced horizontal nystagmus with the same direction as head-turning (geotropic) in the supine roll test, of which 119 patients (63.96%) had a weak vertical upward component of nystagmus on the affected and unaffected sides, 57 patients (30.16%) had a vertical downward component of nystagmus on the affected side and/or the unaffected side, and 13 patients (6.88%) had no vertical component of nystagmus on both the sides. The intensity values of the horizontal component on the affected and unaffected sides were 42.14 ± 24.78 (range: 6.26-138.00°/s) and 17.48 ± 10.91°/s (range: 2.40-53.83°/s), with a ratio of 2.96 ± 2.17:1, representing a significant difference (p < 0.001). We analyzed the characteristics of horizontal and vertical components in 119 patients with HSC-Can (57 L-HSC-Can and 62 R-HSC-Can) on the supine roll test. The intensity values of the horizontal component on the affected and unaffected sides were 43.17 ± 23.76 (range: 8.60-124.51°/s) and 17.98 ± 10.99°/s (range: 2.40-53.83°/s), and the intensity values of the vertical component on the affected and unaffected sides were 10.65 ± 8.46 (range: 1.90-50.83°/s) and 4.81 ± 3.45°/s (range: 0.30-20.43°/s), representing a significant difference between groups (p < 0.001). Among 119 patients with HSC-Can who had a vertical upward component on both the affected and unaffected sides in the supine roll test, unilateral weakness (UW) was normal in 53 cases and abnormal in 51 cases, and 15 cases did not undergo the caloric test. We compared the horizontal and vertical components of nystagmus induced on the affected and unaffected sides in the supine roll test in 53 patients with normal UW and 51 patients with abnormal UW, and the difference was not statistically significant. Conclusion: There is mostly a vertical upward component based on the horizontal component in HSC-Can, and the direction and intensity characteristics of nystagmus accord with Ewald's first law, which can provide a basis for the study of human HSC function and structure, objective diagnosis, and treatment of BPPV.

[Three-dimensional characteristics of torsional nystagmus in induced by posterior semicircular canals canalithasis].

Objective:The three-dimensional direction feature of torsional nystagmus induced by posterior semicircular canal canalithasis （PSC-Can） was recorded and analyzed using three-dimensional video nystagmography （3D-VNG）. Methods:Sixty patients （22 on the left side and 38 on the right side） with PSC-Can were enrolled for torsional nystagmus evoked by Dix-Hallpike test in the affected-side head-hanging and sitting positions, and the direction characteristics of the horizontal, vertical and torsional components were analyzed. Results:Vertical torsional nystagmus was induced in 60 PSC-Can patients in the head-hanging and sitting positions evoked by Dix-Hallpike test, respectively. Horizontal, vertical, and torsional components of were presented in the 3D-VNG. In the head-hanging position, the direction of horizontal component in the left/right PSC-Can nystagmus was contralateral in 46 cases（the other 14 cases were ipsilateral）, the vertical component was upward, and the torsional component was upward/downward, respectively. The intensity of nystagmus induced in the three components in the sitting position is weaker than in the head-hanging position, and the direction of nystagmus was reversed in both vertical and torsional components compared with the head-hanging position. However, the direction of the horizontal component was reversed in 39 cases and not reversed in 21 cases in the sitting position. Conclusion:The horizontal, vertical and torsional components of the torsional nystagmus in PSC-Can patients recorded by 3D-VNG, which provided more comprehensive and objective information for the analysis of PSC-Can and the study of semicircular canal physiological function.

Erythropoietic properties of human induced pluripotent stem cells-derived red blood cells in immunodeficient mice.

Transfusion of red blood cells (RBCs) is a life-saving intervention for anemic patients. Human induced pluripotent stem cells (iPSC) have the capability to expand and differentiate into RBCs (iPSC-RBCs). Here we developed a murine model to investigate the in vivo properties of human iPSC-RBCs. iPSC lines were produced from human peripheral blood mononuclear cells by transient expression of plasmids containing OCT4, SOX2, MYC, KLF4, and BCL-XL genes. Human iPSC-RBCs were generated in culture supplemented with human platelet lysate, and were CD34- CD235a+ CD233+ CD49dlow CD71low ; about 13% of iPSC-RBCs were enucleated before transfusion. Systemic administration of clodronate liposomes (CL) and cobra venom factor (CVF) to NOD scid gamma (NSG) mice markedly promoted the circulatory survival of human iPSC-RBCs following transfusion. While iPSC-RBCs progressively decreased with time, 90% of circulating iPSC-RBCs were enucleated 1 day after transfusion (CD235a+ CD233+ CD49d- CD71- ). Surprisingly, human iPSC-RBCs reappeared in the peripheral circulation at 3 weeks after transfusion at levels more than 8-fold higher than at 1 h after transfusion. Moreover, a substantial portion of the transfused nucleated iPSC-RBCs preferentially homed to the bone marrow, and were detectable at 24 days after transfusion. These results suggest that nucleated human iPSC-derived cells that homed to the bone marrow of NSG mice retained the capability to complete differentiation into enucleated erythrocytes and egress the bone marrow into peripheral blood. The results offer a new model using human peripheral blood-derived iPSC and CL/CVF-treated NSG mice to investigate the development and circulation of human erythroid cells in vivo.

Comprehensive Identification of Bridge Genes to Explain the Progression from Chronic Hepatitis B Virus Infection to Hepatocellular Carcinoma.

BACKGROUND: Hepatitis B virus infection co-occurs in 33% of individuals with hepatocellular carcinoma worldwide. However, the molecular link between hepatitis B virus and hepatocellular carcinoma is unknown. Thus, we aimed to elucidate molecular linkages underlying pathogenesis through in-depth data mining analysis. MATERIALS AND METHODS: Differentially expressed genes were identified from patients with chronic hepatitis B virus infection, hepatocellular carcinoma, or both. Gene set enrichment analysis revealed signaling pathways involving differentially expressed genes. Protein-protein interaction networks, protein crosstalk, and enrichment were analyzed to determine whether differentially expressed gene products might serve as a bridge from hepatitis B virus infection to hepatocellular carcinoma pathogenesis. Prognostic potential and transcriptional and post-transcriptional regulators of bridge genes were also examined. RESULTS: We identified vital bridge factors in hepatitis B virus infection-associated hepatocellular carcinoma. Differentially expressed genes were clustered into modules based on relative protein function. Signaling pathways associated with cancer, inflammation, immune system, and microenvironment showed significant crosstalk between modules. Thirty-two genes were dysregulated in hepatitis B virus infection-mediated hepatocellular carcinoma. CPEB3, RAB26, SLCO1B1, ST3GAL6 and XK had higher connectivity in the modular network, suggesting significant associations with survival. CDC20 and NUP107 were identified as driver genes as well as markers of poor prognosis. CONCLUSION: Our results suggest that the sustained inflammatory environment created by hepatitis B virus infection is a risk factor for hepatocellular carcinoma. The identification of hepatitis B virus infection-related hepatocellular carcinoma bridge genes provides testable hypotheses about the pathogenesis of hepatocellular carcinoma.

[A study of the effect of hot and cold gas perfusion sequence on caloric test results].

Objective: To study whether the warm/cold air injection sequence affects the test results in the caloric test, and provide a basis for the specification and quality control of the caloric test. Methods:Video nystagmography and warm and cold air stimulation apparatus were applied for caloric test. Thirty healthy volunteers （60 ears） were divided into two groups of 15 （30 ears） each. The first group was given cold air stimulation followed by warm air stimulation, and the second group was given heat followed by cold. The differences in nystagmus maximal slow phase velocity （SPV）, semicircular canal paresis （CP） and dominant preponderance （DP） were compared between the two groups of subjects under different perfusion sequences of caloric test. Results:The intensity of nystagmus evoked by subjects in group 1 （cold first and then warm） and group 2 （warm first and then cold） were similar. Paired t-test showed that intra-group analysis of the SPV values of the two groups, comparison of the intensity of nystagmus evoked by different temperatures of the same ear or different sides of ear with the same temperature, the difference was not statistically significant （all P>0.05）. Independent samples t-test showed that between-group analysis of SPV values of two groups, the intensity of nystagmus induced by the same and different temperature stimuli in the ipsilateral ear, the difference was not statistically significant （all P>0.05）. Independent samples t-test showed that the CP values of the two groups were analyzed between groups, and the difference was not statistically significant （all P>0.05）. Independent samples t-test showed that DP values of both groups were in the normal range and the difference was not statistically significant （all P>0.05）. Conclusion:Different perfusion sequences of warm and cold air do not affect the results of caloric tests, and the order of warm and cold air stimulation is not the normative and quality control research direction of caloric test.

Facile synthesis of carboxymethyl cellulose sulfur quantum dots for live cell imaging and sensitive detection of Cr(VI) and ascorbic acid.

Nowadays the synthesis of stable fluorescent sulfur quantum dots (SQDs) remains a big challenge. Herein, the utilization of carboxymethyl cellulose (CMC) to synthesis of SQDs is reported. Benefiting from the unique composition and structure of CMC macromolecule, the resulted CMC-SQDs simultaneously show high aqueous dispersibility and stability, tunable emission, stable fluorescence and low cytotoxicity, which make them promising for working as a fluorescent probe. Fluorescence detection experiments suggested that the CMC-SQDs could serve as a fluorescence on-off-on switch to sensitive and selective detection of Cr(VI) and ascorbic acid (AA) based on the inner filter effect (IFE). The limit of detection towards Cr(VI) and AA can reach 0.024 and 0.18 µM with linear range of 0.5-225 and 1-300 µM, respectively, which compares favorably to other reported fluorescent probes. In addition, the employment of fluorescent CMC-SQDs for practical detection of Cr(VI) and AA was also studied.

[Study on normal reference range of smooth tracking gain for healthy young people].

Objective:Analysis of normal reference value of smooth pursuit test for normal young people. Method:Thirty normal young volunteers were tested by Synapsys videonystagmography. The maximum horizontal tracking angle was 30 °, the vertical maximum tracking angle was 20°, and the frequency was 0.30 Hz, 0.45 Hz and 0.60 Hz, respectively， and the gain under different conditions is used as the observation index. Result:When the frequency is 0.3 Hz, 0.45 Hz, 0.60 Hz, the left and right horizontal gain is 0.92±0.07/0.93±0.07, 0.87±0.08/0.88±0.11, 0.79±0.11/0.78±0.13, respectively, and the asymmetry of left/right gain is 0.021±0.017, 0.031±0.026, 0.037±0.040; the up and down vertical gain is 0.82±0.16/0.80±0.16, 0.78±0.17/0.72±0.15, 0.68±0.20/0.61±0.15, and the asymmetry of the upper/lower gain is 0.046±0.045, 0.069±0.058, 0.109±0.076. Comparing and analyzing the paired left and right gain values of the three frequencies by paired t test, the differences were not statistically significant （P>0.05）. Paired t -test of gain value for different frequency of up and down stationary tracking, the difference was not statistically significant at 0.30 Hz（P>0.05）, but the gain at 0.45 Hz and 0.60 Hz has significant difference（P<0.05）. Comparing the gains of different frequencies in the same direction, the differences in analysis of variance were statistically significant（P<0.05）. Conclusion:The gain value of smooth pursuit test for normal young people can be affected by tracking frequency and direction. At the same frequency, the left/right tracking of 3 frequencies and the up/down tracking gain values of 0.30 Hz are symmetrical, but at 0.45 Hz and 0.6 Hz, the up tracking gain is greater than the down tracking gain, and the gain value in the same direction gradually decreases with the increase of frequency, is the clinical smooth pursuit test is mainly based on 0.30 Hz.