Objective Zinc finger protein 335 (Zfp335) plays a crucial role in the early development of thymic T cells and the differentiation of peripheral T cell subpopulations. The objective of this study is to investigate the role and underlying mechanisms of Zfp335 in the regulation of regulatory T cell (Treg) within tumor immunity. Methods The Zfp335 gene was specifically knocked out in Treg using tamoxifen (Zfp335^fl/fl FOXP3^creERT2), and the MC38 tumor model was established. On the 7th day after tumor inoculation, tumor size was observed and measured. Tumor size was monitored and recorded daily starting from day 7 post-inoculation. On day 12, tumors were harvested, and the proportions of CD4⁺ T cells, CD8⁺ T cells, and Treg were analyzed by flow cytometry. Additionally, the mitochondrial function of effector regulatory T cell (eTreg) was assessed. Results From day 10 post-tumor inoculation, tumor volume in the Zfp335^CKO group was significantly reduced compared to that of the wild-type (WT) group. Furthermore, the infiltration of CD4⁺ and CD8⁺ T cells, along with their respective effector cells, was significantly higher in the Zfp335^CKO group than in the WT group. The proportions of CD4⁺ and CD8⁺ T cells producing interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) were also significantly increased in the Zfp335^CKO group compared to that of the WT group. In addition, the percentage of CD8⁺ T cells secreting granzyme B (GzmB) was significantly higher in the Zfp335^CKO group than that in the WT group. In contrast, the proportion of Treg and inducible T cell co-stimulator (ICOS)⁺ Treg in the Zfp335^CKO group was significantly lower than that in the WT group. Finally, the expression level of Mitotracker Deep Red in eTreg from the Zfp335^CKO group was significantly reduced compared to that in the WT group. Conclusion During tumorigenesis, the specific deletion of Zfp335 impairs Treg activation, which is related to decreased mitochondrial function in eTreg. In Zfp335^CKO mice. Tumors exhibit increased infiltration of effector T cells, accompanied by elevated levels of cytotoxic cytokines, ultimately enhancing resistance to tumor progression.
Animals ; T-Lymphocytes, Regulatory/metabolism* ; Mice ; CD8-Positive T-Lymphocytes/immunology* ; Neoplasms/genetics* ; Cell Line, Tumor ; Mice, Inbred C57BL ; Mice, Knockout ; DNA-Binding Proteins/genetics* ; Female

Objective To screen the active components of total flavonoid extracts of Sarcandra glabra to promote megakaryocyte differentiation.Methods(1)A model of megakaryocyte differentiation disorder was established by co-culturing human megakaryocytic leukaemia cells(Dami)with human bone marrow stromal cells(HS-5)as an evaluation system,and the experimental groupings were as follows:the Dami group(Dami),the control group(Dami+HS-5),and the PMA group[Dami+HS-5+5 ng·mL-1 foprolol 12-tetradecanoate 13-acetate(PMA)],and model group[Dami+HS-5+1%rabbit anti-rat platelet serum(APS)+5 ng·mL-1 PMA]were cultured for 48 hours.The expressions of megakaryocyte differentiation and maturation surface marker molecules,CD41a and CD61 were detected by flow cytometry.(2)Forty-nine SD male rats were randomly divided into blank plasma group,15-minute group,30-minute group,60-minute group,90-minute group,120-minute group,and 240-minute group,with 7 rats in each group.The rats in each administration group were gavaged with 1.26 g·kg-1 of total flavonoids extracts of Sarcandra glabra,and blood was collected at six set time points(15,30,60,90,120,240 minutes)for the preparation of time-dependent serum-containing plasma of total flavonoids extracts of Sarcandra glabra.(3)Ultra-high performance liquid chromatography-quadrupole tandem time-of-flight mass spectrometry(UHPLC-Q-TOF/MS)was used to analyze the plasma of the time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra,and the peak area was used to construct a matrix(X-matrix)of the amount of chemical composition change over time in the time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra.The collected time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra at six different time points was used to intervene in the model of megakaryocyte differentiation and maturation disorder,and the expression of cell surface molecules CD41a and CD61 was detected by flow cytometry to construct the matrix of effect of time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra(Y-matrix).(4)After the data of X and Y matrices were standardized,partial least squares(PLS)was used to calculate and analyze the quantitative and qualitative effect relationship,and variable importance for projection(VIP)>1 was used as the threshold to screen the effect components related to the changes of cell surface molecules CD41a and CD61,and chemical composition identification,as the potential effector components in the total flavonoid extracts of Sarcandra glabra were used to promote the differentiation of megakaryocytes,and finally the regression evaluation system was used to verify the efficacy of its medicinal effect.Results(1)Compared with the Dami group,the expression level of CD41a on the surface of Dami cells in the control group was significantly increased(P＜0.05).Compared with the control group,the expression levels of CD41a and CD61 on the surface of Dami cells in the PMA group were significantly increased(P＜0.01).Compared with the PMA group,the expression levels of CD41a and CD61 on the surface of Dami cells in the model group were significantly reduced(P＜0.01).(2)Compared with the blank plasma group,the expression levels of the molecules CD41a and CD61 on the surface of Dami cells at each time point of 15,30,60,90,120,and 240 minutes were significantly increased(P＜0.01),and the expression levels of CD41a and CD61 were both highest in the 30-minute group.The potential effective components with VIP value greater than 1 were screened out in the positive and negative ion mode,and 540.3638@12.25 and 559.2991@11.53 were selected for pharmacodynamic verification.559.2991@11.53 was identified as daucosterol(Dau),540.3638@12.25 was identified as rosmarinic acid 4-O-β-D-glucoside(Ros).After Ros and Dau intervened in the megakaryocyte differentiation and maturation disorder model respectively,the expression levels of CD41a and CD61 on the surface of Dami cells in the low-,medium-and high-dose groups(40,60 and 80 μg·mL-1)of Ros and Dau were significantly increased compared with the model group(P＜0.05,P＜0.01).Conclusion Ros and Dau may be the active components of the total flavonoids extracts of Sarcandra glabra to promote the differentiation of megakaryocytes.

Objective To investigate the clinical application value of the non-shared incentive diffusion weighted imaging(ZOOM-DWI)technique at cervical level in diagnosis of cervical spondylotic myelopathy(CSM).Methods A total of 49 CSM patients(patient group),and 50 healthy volunteers(control group)were recruited.All subjects underwent conventional MRI and ZOOM-DWI of the cervical spine and neurologic modified Japanese Orthopaedic Association(mJOA)scores in patients with CSM.The apparent diffusion coefficient(ADC)value in the spinal cord at the narrowest area(C5-C6)of the compression site of patients,the ADC value at the disc level in each upper and lower level,and the spinal ADC value at the cervical level C2-C3 were measured.The ADC values of control group C2-C3,C3-C4,C4-C5,C5-C6,C6-C7 were measured.Within-group comparisons of the spinal cord ADC values for each segment between patient and control groups were performed using analysis of variance and post hoc comparisons(SNK-q).The ADC values at the narrowest point of the patient group and control group were tested by independent sample t-test.The Pearson correlation analysis was performed between patients'C5-C6 ADC values and mJOA scores.Results The mean ADC values showed no significantly different levels in the control group.Among the ADC values at each measurement level in the patient group,except for C4-C5 and C6-C7 where the difference was not statistically significant,the remaining pair-wise comparisons all showed statisti-cally significant differences(F=24.368,P＜0.001),with the highest ADC value at C5-C6.The C5-C6 ADC value in the patient group was significantly higher compared to the control group(t=9.414,P＜0.001),with statistical significance.The ADC values at the patient stenosis showed a significant negative correlation with the mJOA score(r=-0.493,P＜0.001).Conclusion Cervical ZOOM-DWI technique can be applied to diagnose CSM,and spinal ADC values can be used as reliable imaging data for diagnosing CSM.

OBJECTIVE To study the impr ovement effects of Dianxianqing granule on blood-brain barrier （BBB）injury in Alzheimer’s disease （AD）model mice by regulating NLR family pyrin domain containing 3（NLRP3）inflammasome signaling pathway. METHODS Totally 125 mice were randomly divided into sham operation group （n＝25）and modeling group （n＝100） by body weight. AD model was induced by intracerebroventricular injection of β-amyloid 25-35 in model group. Sham operation group was given normal saline with same method. The 100 model mice were randomly divided into model group ，Donepezil hydrochloride tablets group （positive control 1，1.3 mg/kg，i.g.），MCC950 group [positive control 2（selective NLRP 3 inhibitor），10 mg/kg，i.p.] and Dianxianqing granule group （12.48 g/kg by crude drug ，i.g.）by body weight ，with 25 mice in each group. Second day after modeling ，administration groups were given relevant medicine ，once a day ，for consecutive 21 d. Sham operation group and model group were given intragastric administration of water and intraperitoneal injection of normal saline. At last administration，the learning and memory ability was determined by Y maze test ，and blood-brain barrier permeability was measured by Evans blue leakage assay. The expressions of NLRP 3，anti-ionized calcium-binding adapter molecule 1（IBA-1），nuclear factor kappa B （NF-κB）p65，p53 upregulated modulator of apoptosis （PUMA），occludin（ocln），zonula occluden- 1（ZO-1）and claudin-5 （cldn5） in cerebral tissue were determined. RESULTS Compared with model group ， spontaneous alternate response rate ，protein expressions of ocln ，cldn5 lnzyxyqy2003@163.com and ZO- 1 in cerebral tissue were increased significantly in administration groups （P＜0.05 or P＜0.01）；Evans blue E-mail：jiadg2003@126.com content and protein expressions of NLRP 3，IBA-1，PUMA and NF-κB p65 in cerebral tissue were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS Dianxianqing granule can improve BBB injury of AD model m ice by inhibiting NLRP 3 inflammasome signaling pathway.

Objective:To analyze the postoperative paravertebral muscle degeneration and its correlations with health related quality of life (HRQL) in patients undergoing minimally invasive surgery-transforaminal lumbar interbody fusion (MIS-TLIF).Methods:The clinical data of the 50 patients were retrospectively analyzed who had undergone single-segmental MIS-TLIF at Department of Spinal Cord Surgery, The People's Hospital of Henan Province from January 2019 to December 2021. The relative volumes of lumbar posterior muscle (LM), the relative volumes of the psoas major (PM), and the rates of fatty degeneration (FD) of the fused segment and its adjacent segments were compared respectively between preoperation, 6 and 12 months postoperation. The correlations were analyzed between the HRQL scores [visual analog scale (VAS) for pain and Oswestry disability index (ODI)] and the relative LM volumes, the relative PM volumes, and the FD rates of the fused segment and its adjacent segments at 12 months postoperation.Results:Compared with the preoperative values, the relative LM volumes and the relative PM volumes of the fused segment and its adjacent segments at 6 and 12 months postoperation were significantly reduced while the FD rates significantly increased. However, the FD rate of the fused segment at 12 months postoperation (20.6% ± 6.1%) was significantly lower than that at 6 months postoperation (29.7% ± 8.2%) ( P < 0.05). The VAS score was strongly negatively or positively correlated with the relative LM volume ( r = -0.819, P < 0.001) and the FD rate ( r = 0.86, P < 0.001) of the fused segment, and moderately negatively correlated with the relative PM volume ( r = -0.435, P = 0.016). The ODI index was moderately negatively correlated with the relative LM volume ( r = -0.512, P = 0.004) and the relative PM volume ( r = -0.402, P = 0.020) of the fused segment, but moderately positively correlated with the FD rate of the fused segment ( r = 0.565, P = 0.001). There was a moderate negative correlation between the ODI index and the relative LM volume of the adjacent segments ( r = -0.478, P = 0.012). Conclusions:After MIS-TLIF, the volume of the paravertebral muscles decreases and the dorsal muscles develop fatty degeneration. The improvement of LM fatty degeneration may be observed by 12-month follow-up in the fused segment, but not in the adjacent segments. The LM volume and the FD rate of the fused segment are the most closely related to the postoperative HRQL.

OBJECTIVE To study the improvement effects of Huorongcaohuangqi （HRCHQ） admixture decoction on mesangial proliferative glomerulonephritis （MsPGN）model rats and its mechanism . METHODS Totally 70 rats were divided into sham operation group （n＝10）and modeling group （n＝60）according to body weight . Sham operation group underwent sham operation，and MsPGN model was induced by immunological method （Freund’s adjuvant+BSA+lipopolysaccharide ）in modeling group. After successfully modeling ，50 rats were randomly divided into model group ，HRCHQ high -dose，medium-dose and low - dose groups （4.05，2.03，1.02 g/kg），Benazepril hydrochloride tablet group （20 mg/kg），with 10 rats in each group . Sham operation group and model group were given distilled water intragastrically ；other groups were given relevant medicine intragastrically 15 mL/kg，once a day ，for consecutive 5 weeks. After last administration ，the contents of total protein ，albumin and urea nitrogen were determined in the serum of rats . The expressions of transforming growth factor β1（TGF-β1）and tumor necrosis factor α（TNF-α）in renal tissue were detected by immunohistochemistry ；the expression of complement C 3 in renal tissue was detected by immunofluorescence . The phosphorylation level of phosphatidylinositol 3 kinase/protein kinase B/glycogen synthase kinase 3β（PI3K/Akt/GSK-3β） signal pathway related proteins and expression level of fibronectin （FN） in renal tissue were detected by Western blot . RESULTS Compared with model group，the serum contents of total protein and albumin were increased significantly in HRCHQ high -dose group （P＜0.05 or P＜0.01）. The serum content of urea nitrogen ， the E-mail：Houxia_75@163.com expressions of TGF -β1，TNF-α，FN and complement C 3，and the phosphorylation levels of PI 3K，Akt and GSK 3β protein were all decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS HRCHQ can improve MsPGN by reducing the proliferation of mesangial cells and inhibiting the activity of PI 3K/ Akt/GSK3β signaling pathway .

Objective:To analyze the sleep quality and sleep structure of patients with obstructive sleep apnea hypopnea syndrome (OSAHS) complicated with patent foramen ovale (PFO), and to study the effect of PFO on the sleep structure of OSAHS.Methods:Fifty-six patients with OSAHS complicated with PFO, 64 patients with simple OSAHS and 62 controls were collected from December 2018 to March 2020 in Centre of Sleep Disorders, the Second Affiliated Hospital of Zhengzhou University. Pittsburgh Sleep Quality Index and polysomnography were used to compare the sleep quality and sleep structure of the three groups.Results:Compared with the control group [6/62(9.68%)], OSAHS complicated with PFO group [54/56(96.43%)] and simple OSAHS group [53/64(82.81%)] had higher incidence of poor sleep quality (χ2=112.08, P<0.0l). Furthermore, compared with the control group, the OSAHS complicated with PFO group and simple OSAHS group showed reduced sleep efficiency [PSQI total score was 0.5 (0, 1), 2 (1, 3) and 2 (1, 2) respectively, H=74.549, P<0.01] and reduced proportions of rapid eye movement (REM; 20.45%±3.49%, 12.19%±5.95% and 15.11%±7.21%,respectively, F=21.17, P<0.01) and slow wave sleep (N3; 21.24%±4.12%, 14.15%±6.08%, 17.68%±6.35%, respectively, F=29.51, P<0.01); the N1 (4.47%±2.40%, 9.50%±5.34%, 9.55%±4.61%, respectively, F=30.07, P<0.05) and N2 sleep (53.88%±4.35%, 64.09%±7.49%, 58.14%±6.67% , respectively, F=46.21, P<0.05) were prolonged; the inocturnal lowest oxyhemoglobin saturation (SpO 2) level was lower, mean SpO 2 reduction at night was higher [3.00% (0, 4.00%),6.00% (5.00%, 8.75%) and 4.00% (4.00%, 5.00%), respectively, H=72.24, P<0.05], and periodic leg movement index [16.30(4.80, 32.82), 33.30(9.26, 54.80) and 23.10(8.38, 31.83),respectively, H=17.86, P<0.05], arousal index [11.60(7.73, 17.55), 23.90(14.03, 30.45) and 15.6(11.23, 20.78), respectively, H=22.80, P<0.05] and sleep apnea and hypopnea index (AHI; 1.60±1.38, 23.90±7.27 and 16.24±4.22,respectively, F=136.97, P<0.05) increased. Compared with the simple OSAHS group, the incidence of poor sleep quality was higher, the proportions of slow wave sleep (N3, F=29.51, P=0.047) and REM ( F=21.17, P=0.012) were decreased, N2 sleep ( F=46.21, P=0.000) was prolonged, mean SpO 2 reduction at night ( Z=54.28, P=0.000), wake after sleep onset [116.00(89.88, 143.00) min vs 135.00(118.50, 168.38) min, Z=25.71, P=0.023], arousal times [14.00(8.25, 8.00) vs 17.50(9.00,23.00),respectively, Z=19.68, P=0.041], microarousal ( Z=23.57, P=0.044), and AHI ( F=136.97, P=0.000) were increased in the OSAHS complicated with PFO group. Conclusions:OSAHS complicated with PFO patients had poor sleep quality and high incidence of sleep disorders. They had sleep disorder at night, which was characterized by the decrease of REM sleep and slow wave sleep, the prolongation of N2, the decrease of nocturnal SpO 2 and the increase of awakening times, and the increase of arousal times and AHI. PFO can aggravate the sleep disorder of OSAHS.

Objective:To quantitatively evaluate myocardial microcirculation perfusion changes after percutaneous intracardiac septal radiofrequency ablation (PIMSRA, Liwen procedure) in patients with obstructive hypertrophic cardiomyopathy (HOCM) using myocardial contrast echocardiography (MCE) and to further establish the effect of the procedure and predict the long-term prognosis of patients.Methods:A total of 45 patients with HOCM treated by the Liwen procedure in the First Affiliated Hospital of Air Force Medical University (Xijing Hospital) from July 2019 to June 2020 were included. MCE was performed before and 6 months after surgery, respectively. Time-intensity perfusion curve analysis was performed using QLab 10.8 offline software to obtain quantitative parameters of myocardial microcirculation perfusion including myocardial blood volume (A value), myocardial blood flow velocity (β value), and myocardial blood flow (A×β value), then the changes in parameters before and after the procedure were compared.Results:After the Liwen procedure, the mean septal thickness and mean left ventricular free wall thickness were significantly reduced, accompanied by a significant reduction in the left ventricular outflow tract pressure gradient (LVOT-PG) and mitral regurgitation length, and a significant improvement in mitral systolic antegrade motion (SAM) (all P<0.001). In addition, the left ventricular ejection fraction (LVEF) did not deteriorate significantly ( P=0.560) and the E/e′ ratio decreased after the procedure ( P=0.015). Besides, the A values of both the ventricular septum and the left ventricular free wall were not significantly changed compared to those before procedure ( P>0.05), whereas the β values and A×β values were obviously increased ( P<0.05). Conclusions:Myocardial blood velocity and myocardial blood flow in the septum and left ventricular free wall were significantly increased in patients with HOCM after the Liwen procedure, suggesting a significant improvement in myocardial microcirculation perfusion. MCE provides a non-invasive quantitative evaluation parameters of myocardial microcirculation perfusion for the Liwen procedure for the treatment of HOCM.

Objective:To investigate the feasibility of ultrasound-guided percutaneous endomyocardial septal cryoablation of in vitro porcine hearts and to compare its effect with the percutaneous endomyocardial radiofrequency ablation.Methods:Experiment 1: Six in vitro porcine hearts were divided into 1 min ( n=2), 3 min ( n=2) and 5 min ( n=2) groups according to the cryoablation time, and all were subjected to ultrasound-guided percutaneous intra-myocardial septal cryoablation at 100% power respectively. After cryoablation, ultrasound images, the size of the solid dissection of the ice ball, and the size of the necrotic area after melting of the frozen ice ball were measured. Experiment 2: The in vitro porcine hearts were divided into cryoablation group ( n=3) and radiofrequency ablation group ( n=3), and ultrasound-guided percutaneous endomyocardial septal cryoablation and radiofrequency ablation were performed with 100% cryo power and 40 W radiofrequency power, and the extent of complete necrotic area and incomplete necrotic area were compared between the two ablation methods after 1 min. Results:Experiment 1: In the 1 min cryoablation time group ( n=2), the short diameter of the puck measured by ultrasound was (8.00±0.84)mm, the short diameter of the puck measured by solid was (8.38±1.19)mm, and the short diameter of the necrotic zone measured by solid was (8.35±0.83)mm; in the 3 min group ( n=2), the short diameter of the puck measured by ultrasound was (19.4±0.28)mm, and the short diameter of the puck measured by solid was (19.03±0.33)mm, solid measurement of the short diameter of the necrotic zone was (19.16±0.25)mm; in the 5 min group ( n=2), the short diameter of the puck measured under ultrasound was (26.4±2.54)mm, solid measurement of the short diameter of the puck was (26.01±0.24)mm, and solid measurement of the short diameter of the necrotic zone was (24.82±0.25)mm. Randomized blocks analysis of variance was performed on this data and the difference of block Factor b (freezing time: 1 min, 3 min, 5 min) among the three groups was statistically significant( F=505.884, P<0.001). The SNK- q test showed that all three groups differed from each other(all P<0.05). The analysis results for the treatment factors K (measurement modality-ultrasound image measurements, solid anatomical measurements of the puck, and measurements of the necrotic area after melting of the frozen puck) was not statistically significant ( F=0.470, P=0.635). Experiment 2: In the RF ablation group ( n=3), the ratio of incomplete necrotic zone to the radius of the RF ablation area was 0.64±0.01; in the cryoablation group ( n=3), the ratio of incomplete necrotic zone to the radius of the ablation area was 0.26±0.02. The difference was statistically significant( P=0.002) and it can be considered that the incomplete necrotic zone of cryoablation was smaller than that of RF ablation. Conclusions:Percutameous intramyocardial septal cryoablation is controllable in scope, ultrasound image evaluation of ablation area is more accurate and incomplete necrosis area is small, which may have potential applications in cardiac ablation.

Objective: To perform a comparative study of different fat-processing techniques to explore which technique could obtain higher fat content. Methods: Lipoaspirates was purified with three groups of fat-processing techniques (sedimentation, centrifugation, and cotton pad filtering). Isometric purified fat tissue was centrifuged at 5 000 g for 5 minutes to quantify the oil, aqueous, and fat content from them. HE staining was applied to evaluate the effect of different fat-processing techniques on the integrity of adipocyte. Results: The fat content, aqueous content, and oil content of three groups was respectively 86.0%, 9.6% and 4.4% for the cotton pad filtering group; 74.4%, 14.8% and 10.8% for the centrifugation group; 62.0%, 28.4% and 9.6% for the sedimentation group. In the HE histologic findings, no significant difference was observed in the integrity of adipocyte of three groups (P>0.05). Conclusions In comparison with the sedimentation group and the centrifugation group, more fat content, less oil content and aqueous content was observed in the cotton pad filtering group. The cotton pad filtering technique was helpful in postoperative volume retention.