[Objective] To explore the feasibility of using whole blood as a source of NK cells for allogeneic CAR NK cell therapy and activated NK cell reinfusion therapy, and initially construct a technical system for the separation and purification of NK cells from whole blood. [Methods] All peripheral blood mononuclear cells (PBMCs) were enriched from 400 mL of whole blood by manual separation and machine separation, respectively. The erythrocyte loss rate, PBMCs number, NK cell purity of the two methods were compared. NK cells were sorted from PBMCs by three separation and enrichment methods as immunomagnetic bead negative selection method, platelet lysate culture expansion and PERCOLL density gradient separation method, and the purity and yield of NK cells, the activity of NK cells and the tumor-killing ability of the three separation and enrichment methods were compared. [Results] The proportion of NK cells in the lymphocyte population was higher in the manual separation method than in the machine separation method[(13.16±5.16)% vs (8.56±3.92)%, P<0.05]; the number PBMCs was lower in the manual separation method than in the machine separation method[(4.09±1.80)×10⁸vs (6.49±2.16)×10⁸, P<0.05], and there was no difference in the red blood cell loss between the two methods (P>0.05). The purity of NK cells isolated and enriched from PBMCs by manual separation method using immunomagnetic was (96.77±2.31)%; the yield was (56.27±10.47)%; the inhibition of tumor proliferation was (38.67±14.05)%; and the tumor killing rate was (19.90±8.05)%. The purity of NK cells isolated and enriched from PBMCs by manual separation method using platelet lysis culture expansion method was the highest at day 7, which was (54.84±15.80)%; the cell expansion multiple could reach 16.92±6.28 at day 7; the in vitro tumor killing rate of NK cells was (15.83±5.5)%; the tumor inhibition rate was (44.33±13.5)%; and there was no difference in the toxicity and activity of NK cells between the two methods (P>0.05). The purity of NK cells isolated and enriched by PERCOLL density gradient separation method was (15.83±5.82)%, and the yield was (14±6.25)%, which was significantly lower than the other two methods. [Conclusion] PBMCs isolated from whole blood by manual separation and NK cells enriched by negative selection with immunomagnetic beads have the potential to provide NK cell materials for CAR-NK cell therapy, and NK cells enriched by platelet lysate-conditioned medium have the potential to provide NK cells for large-scale NK cell activation reinfusion therapy.

ObjectiveTo investigate the triglyceride-glucose (TyG) index and the level of serum homocysteine (Hcy) in association with the incidence of stroke in type 2 diabetes mellitus (T2DM) patients. MethodsBased on the chronic disease risk factor surveillance cohort in Pudong New Area, Shanghai, excluding those with stroke in baseline survey, T2DM patients who joined the cohort from January 2016 to October 2020 were selected as the research subjects. During the follow-up period, a total of 318 new-onset ischemic stroke patients were selected as the case group, and a total of 318 individuals matched by gender without stroke were selected as the control group. The Cox proportional hazards regression model was used to adjust for confounding factors and explore the serum TyG index and the Hcy biochemical indicator in association with the risk of stroke. ResultsThe Cox proportional hazards regression results showed that after adjusting for confounding factors, the risk of stroke in T2DM patients with 10 μmol·L⁻¹-¹ and Hcy>15 μmol·L⁻¹ was 1.532 times (95%CI: 1.017‒2.309 times, P=0.041) and 1.738 times (95%CI: 1.119‒2.699 times, P=0.014) higher respectively, compared to T2DM patients with Hcy≤10 μmol·L⁻¹. T2DM patients with TyG>9.074 had 1.396 times higher risk of stroke (95%CI: 1.091‒1.787, P=0.008) compared to those with TyG≤9.074. The study found that urea and α1-antitrypsin (α1-AT) were independent risk factors for the incidence of stroke in T2DM patients. For every unit increase in urea andα1-AT, the risk of stroke in T2DM patients increased by 233.6% (HR=3.336, 95%CI: 1.588‒7.009, P=0.001) and 11.3% (HR=1.113, 95%CI: 1.028‒1.205, P=0.008), respectively. Cumulative risk curves showed that Hcy>10 μmol·L⁻¹ and TyG>9.074 accelerated the time to stroke occurrence in T2DM patients. ConclusionElevated levels of Hcy>10 μmol·L⁻¹ and a higher TyG index are risk factors for stroke in T2DM patients. Effective interventions for Hcy and TyG should be conducted for diabetic patients to reduce the incidence of stroke.

ObjectiveTo investigate the correlation between iron metabolism parameters and various syndrome types of unstable angina pectoris （UAP）. MethodsA cross-sectional study was conducted from October 2021 to October 2023， encompassing 213 patients diagnosed with UAP at Xiyuan Hospital of Chinese Academy of Chinese Medical Sciences. Additionally， 30 healthy individuals were selected as control cases. Single-factor analysis was used to investigate the differences in clinical data among different Traditional Chinese Medicine （TCM） syndrome types of UAP and their correlation with iron metabolism indices. The study conducted a comparative analysis of the aforementioned clinical data among patients with and without heat-toxic and blood-stasis syndrome. Logistic regression was used to analyze the correlation between TCM syndrome types and related factors. The receiver operator characteristic （ROC） curve was employed to assess the predictive value of iron metabolism indices， along with their sensitivity and specificity. ResultsCompared to those in the control group， serum iron （SI） and serum ferritin （SF） levels were significantly increased in the UAP group （P<0.01）， while transferrin （TRF） and total iron binding capacity （TIBC） levels were decreased （P<0.01）. However， there was no significant difference in unsaturated iron binding capacity （UIBC）. Multivariate binary Logistic regression analysis identified apolipoprotein A1 （ApoA1）， homocysteine （HCY）， high-sensitivity C-reactive protein （hs-CRP）， and SF as independent influencing factors for the UAP patients （P<0.05， P<0.01）. Additionally， statistically significant differences were observed in SI， SF， TRF， and TIBC among 213 patients with different TCM types （P<0.01）. Patients with heat-toxic and blood-stasis syndrome had higher SI and SF values than those without the syndrome （P<0.01）， while their TIBC and TRF values were lower （P<0.01）. Multivariate binary logistic regression analysis showed that SI and LDL-C levels were closely associated with the differentiation of heat-toxic and blood-stasis syndrome. ConclusionUAP patients often experience iron metabolism disorders， and the heat-toxic and blood-stasis syndrome are significantly correlated with iron metabolism parameters. The SI and LDL-C levels have high specificity and sensitivity in diagnosing heat-toxic and blood-stasis syndrome.

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 explore the clinical characteristics, audiological outcomes, and factors influencing the efficacy of pharmacological treatment in patients with sudden hearing loss associated with large vestibular aqueduct syndrome（LVAS）. Methods:A retrospective analysis was conducted on the clinical data of 77 bilateral LVAS patients（117 ears） hospitalized for sudden hearing loss from January 1, 2009, to December 31, 2023. The inclusion criteria required that patients to be diagnosed according to the Valvassori standard and had received standardized pharmacological treatment. Clinical features, audiological outcomes, and treatment efficacy were analyzed. Statistical methods were employed to identify factors associated with treatment outcomes. Results:The age of the enrolled patients ranged from 4 to 37 years. The age of onset for the initial hearing fluctuation varied between 0 and 24 years, with a mean age of 5.8 years. The male-to-female ratio was approximately balanced（37 males and 40 females）. The proportion of unilateral to bilateral sudden hearing loss was 1.0︰1.2, with unilateral right ear hearing loss being more frequently occurring（64.9%）. Triggering Factors: Triggers included no identifiable factors in 48.1% of cases, a history of head trauma（24.7%）, upper respiratory tract infections（11.7%）, onset following physical fatigue（11.7%）, and less frequently, noise exposure, alcohol consumption, or emotional stress（each 1.3%）. Clinical Symptoms: Hearing loss was the sole symptom in 35.1% of cases. Concurrent symptoms included vertigo in 44.2% and tinnitus in 46.8%. Patients with a disease duration of ≤14 days demonstrated a treatment efficacy rate of 75.0%. Among those who responded to treatment, 93.0% had profound or greater hearing loss prior to therapy, with an average improvement in hearing thresholds of 32 dB HL. In pretreatment, 68.9% of patients exhibited low-frequency air-bone gaps, increasing to 76.1% post-treatment. Additionally, 17.6% of treated ears demonstrated a ≥15 dB HL improvement in low-frequency bone conduction thresholds. In the non-responsive group, 7.3% of ears still showed some improvement in bone conduction thresholds. Statistically significant differences（P<0.05） were observed between the treatment-effective and non-effective groups concerning the age of initial hearing fluctuation, disease duration, and severity of hearing loss at onset. Conclusion:The efficacy of pharmacological treatment for sudden hearing loss in LVAS patients is influenced by the age at onset, duration of the disease, and severity of hearing impairment. Early diagnosis and timely intervention significantly enhance treatment efficacy, particularly in patients with a disease duration of ≤14 days and an initial sudden hearing loss. Patients with severe hearing loss, especially those with profound or greater impairment, exhibit greater sensitivity to treatment. Pharmacological interventions positively impact both air conduction and bone conduction thresholds, with the observed improvement in bone conduction thresholds warranting further investigation.
Humans ; Male ; Retrospective Studies ; Hearing Loss, Sudden/etiology* ; Female ; Vestibular Aqueduct/pathology* ; Adult ; Child ; Adolescent ; Child, Preschool ; Young Adult ; Treatment Outcome

Objective:To explore the clinical effect of rejuvenation of lower eyelid through orbital septum fat and orbital repositioning combined with thermal compression.Methods:A total of 423 patients who underwent skin approach blepharoplasty with skin laxity accompanied by different degrees of lacrimal groove and palpebromalar groove deformity admitted to the Plastic Surgery Department of Hemangioma and Vascular Malformation, the Plastic Surgery Hospital of Chinese Academy of Medical Sciences, from April 2020 to January 2023 were selected, including 403 females and 20 males, aged 33-68 (45.0±11.1) years. During the operation, the combined subcutaneous and suborbicularis oculi approaches were used to separate the prezygomatic space and premaxillary space. In the space, the orbital septum fat flap was fixed with the orbital septum flap under the infraorbital margin by lmbricate suture, and the orbital septum was tightened with energy hemostasis device to reshape and strengthen the anatomical structure of the lower eyelid. The patients were followed up to evaluate the incision healing and complications after surgery.Results:All the 423 patients were followed up for 1-48 months. The patients were healed in one stage, without lower eyelid retraction, lower eyelid ectropion and other complications. The incision of the lower eyelid was concealed and smooth, and the depression of lacrimalgroove and palpebromalar groove and lower eyelid relaxation were improved significantly.Conclusions:For patients with lower eyelid bag skin laxity and different degrees of lacrimal groove and palpebromalar groove depression, the surgical design is reasonable, and the structure of orbital septum fat and orbital septum is reset, and the effect is reliable.

Objective:To evaluate the testing capability of urinary iodine laboratories in Gansu Province and analyze the existing problems.Methods:Z-score method and uncertainty analysis were used to analyze the external quality control assessment results of urinary iodine laboratories in Gansu Province from 2017 to 2021 (data were collected from Gansu Center for Disease Prevention and Control).Results:From 2017 to 2021, the participation rate in the assessment of urinary iodine laboratories in the province was 100.0% (473/473), the feedback rate was 99.8% (472/473), and the pass rate was 91.9% (434/472). The pass rates for assessment from 2017 to 2021 were 82.7% (62/75), 93.9% (93/99), 94.9% (93/98), 92.0% (92/100), and 94.0% (94/100), respectively. The pass rates for provincial, municipal, and county assessments were 5/5, 98.6% (69/70), and 90.7% (360/397), respectively. The proportions of │Z│≤2, 2 <│Z│ < 3, and│Z│≥3 between laboratories in the province were 84.5% (399/472), 9.3% (44/472), and 6.2% (29/472), respectively. The proportions of│Z│≤2, 2 <│Z│ < 3, and│Z│≥3 within the laboratories were 88.6% (418/472), 9.1% (43/472), and 2.3% (11/472), respectively. There was a significant difference in the composition of │Z│ scores between laboratories annually (χ 2 = 24.60, P = 0.002), the proportion of│Z│≤2 increased from 66.7%(50/75) in 2017 to 90.0% (90/100) in 2021. The│Z│ scores between and within provincial laboratories were both ≤2. The proportion of │Z│≤2 between municipal and county-level laboratories was 91.4% (64/70) and 83.1% (330/397), respectively, the proportion of│Z│≤2 within laboratories was 92.9% (65/70) and 87.7% (348/397), respectively. There was no difference in the composition of│Z│ scores between and within provincial, municipal, and county-level laboratories( P < 0.05). The proportion of two concentration quality control blind sample results in the province that were both within the uncertainty range was 89.2% (421/472). From 2017 to 2021, they were 81.3% (61/75), 91.9% (91/99), 84.7% (83/98), 92.0% (92/100), and 94.0% (94/100), respectively, with statistically significant differences (χ 2 = 9.69, P = 0.021); provinces, cities, and counties were 5/5, 95.7% (67/70), and 87.9% (349/397), respectively, with statistically significant differences (χ 2 = 23.60, P = 0.023). Conclusions:Through continuous external quality control assessments of all established urinary iodine laboratories in Gansu Province, the overall testing capacity of urinary iodine laboratories at all levels has been continuously improved. However, in the future, it is still necessary to strengthen laboratory testing capabilities and improve the level of urine iodine detection.

Objective To investigate the long-term effect of frailty on heart failure with reduced ejection fraction(HFrEF)in elderly patients.Methods A retrospective analysis was conducted on 245 HFrEF patients aged ≥75 years admitted to our hospital from October 2017 to October 2020 due to acute exacerbation of chronic heart failure(HF).Based on their clinical frailty scale(CFS)score,they were divided into frailty group(1-4,135 cases)and non-frailty group(5-9,110 cases).Their general clinical data,clinical medication,and prognosis were compared between the two groups,and the influencing factors for frailty and death were analyzed.Results Faster heart rate,higher NT-proBNP level,and larger proportions of male,diabetes,coronary heart disease,≥5 chronic diseases,LVEF ≤35％,anemia and increased troponin I level,while lower BMI,eGFR and score of activity of daily living scale were observed in the frailty group than the non-frailty group(P＜0.05,P＜0.01).The frail group had significantly lower utilization rates of angiotensin converting enzyme inhibitors(ACEI),angiotensin receptor blockers(ARB),angiotensin receptor enkephalin inhibitors(ARNI)β receptor blockers,and sodium-glucose cotransporter 2 inhibitors than the non-frailty group(P＜0.01).Additionally,the frailty group exhibited a higher incidence of emergency room visits/readmissions within 3 months and 2-year mortality than the non-frailty group(P＜0.05,P＜0.01).Binary logistic regression analysis revealed that ≥5 chronic diseases,LVEF ≤ 35％,BMI,and GFR were independent risk factors for frailty(OR=0.167,95％CI:0.064-0.453,P=0.000;OR=0.306,95％CI:0.160-0.586,P=0.000;OR=0.868,95％CI:0.786-0.958,P=0.005;OR=0.966,95％CI:0.943-0.991,P=0.007),while ≥5 chronic disea-ses and frailty were independent risk factors for death in HF patients(P＜0.05).Conclusion The incidence of frailty is high in elderly HF patients with HFrEF.They have poor compliance to guideline directed drug therapy(GDMT).Frailty is an independent risk factor for long-term mor-tality in the patients.

Objective To assess the safety of femoral artery puncture procedures guided by X-ray and ultrasound among elderly patients.Methods A total of 480 patients undergoing transcatheter interventional treatment for cardiovascular and cerebrovascular diseases through the femoral ar-tery in our hospital between January 2016 and December 2022 were enrolled in the study.Of them,326 patients receiving femoral artery puncture guided by X-ray fluoroscopy were assigned into X-ray group,while the other 154 patients guided by vascular Doppler ultrasound were into ultrasound group.With propensity score matching(PSM)in a ratio of 1∶1,finally 270 patients were included.Their general clinical data,success rate of puncture,puncture site,and incidence of vascular complications were compared between the two groups.Multivariate logistic regression analysis was used to identify the risk factors for vascular complications.Results Before PSM,there were no statistical differences in the mean distance from the skin fold to the bifurcation of the common femoral artery(2.5±1.0 cm vs 2.4±0.8 cm)or the distance from the fold to the in-guinal ligament(6.4±1.4 cm vs 6.3±1.7 cm)between the X-ray group and the ultrasound group(P＞0.05).After PSM,the X-ray group exhibited an obviously higher incidence of puncture points below the common femoral artery than the ultrasound group(14.8％vs 6.7％,P＜0.05),but no significant differences were observed in the one-time success rate of puncture or the occur-rence of vascular complications between the two groups(P＞0.05).Multivariate logistic regres-sion analysis indicated that the presences of non-common femoral artery and femoral artery calci-fication at the puncture site was independent risk factors for vascular complications(OR=8.379,95％CI:3.561-19.717;OR=3.922,95％CI:1.664-9.242).Conclusion There is no statistical disparity in safety between X-ray-versus ultrasound-guided femoral artery puncture procedures.Cli-nicians should choose appropriate puncture procedure or combine them together based on individual con-dition of patients.