Objective To investigate the feasibility of granzyme B(GB)promoter-controlled ferritin heavy chain(FTH1)reporter gene expression for monitoring the activation status of chimeric antigen receptor T cells(CAR-T)by magnetic resonance imaging(MRI).Methods Cytotoxic T lymphocytes(CTLs)were screened by Ficoll density gradient centrifugation and flow sorting.The GB promoter and FTH1 gene were ligated together with disialoganglioside 2(GD2)CAR,and lentiviral vectors were transfected into CTLs to construct GD2-CAR-T/pGB-FTH1 cells.GD2-CAR-T/pCMV-FTH1,GD2-CAR-T,and T cells served as control cells.CytoTox96@non-radioactive cytotoxicity was used to detect the killing effect of each group of cells after co-culture with human neuroblastoma cells(SK-N-SH).ELISA was employed to detect the coincubation factor as well as the amount of GB secretion.Western blotting,Prussian blue staining and cellular MRI were applied to detect the expression of the FTH1 gene after co-culture.Results CTLs were successfully obtained,and then GD2-CAR-T/pGB-FTH1,GD2-CAR-T/pCMV-FTH1 and GD2-CAR-T cells were constructed.The killing effect,co-incubation factor and GB secretion of the above 3 groups of cells were significantly higher than those of the T cells,and the level of GB expression was highest at day 1,and then decreased in order at day 3 and day 7 after co-culturing with SK-N-SH cells.The relative expression of FTH1 and iron content of the GD2-CAR-T/pGB-FTH1 cells showed the same trend as GB expression,and the MRI signals were gradually increased.There were no significant differences in the relative expression of FTH1,iron content and MRI signals in the GD2-CAR-T/pCMV-FTH1 cells at all time points.No FTH1 expression or iron aggregation was observed in the GD2-CAR-T and T cells groups.Conclusion MRI based on the FTH1 reporter gene driven by the granzyme B promoter can reflect the GB expression level and tumor killing effect of CAR-T cells,which provides a potential real-time visual means to monitor the cell activation status for CAR-T therapy.

Objective:To analyze the costs and effectiveness of five common screening modes and genetic screening for thalassemia in China in order to find the optimal way and provide evidence for the implementation of thalassemia prevention and control projects in Hunan Province.Methods:From June 2020 to April 2021, 12 971 couples from 14 cities and autonomous prefectures in Hunan Province were selected as the study population. The diagnosis of thalassemia was based on the results of genetic testing. Results of routine blood test and hemoglobin electrophoresis were collected and analyzed. The efficacy of five screening modes, at the cut-off value of <80 fl or 82 fl for the mean corpuscular volume (MCV), was analyzed by positive predictive value, negative predictive value, Jorden index and cost-effectiveness ratio. Sensitivity analysis was used to assess the feasibility of genetic screening at different costs after fixing the costs of routine blood and hemoglobin electrophoresis. The five thalassemia screening models are as follows: Mode 1: The woman had a blood routine test first. If the result was positive, the spouse required a blood routine test. If both results were positive, a thalassemia gene test should be offered to the couple. Mode 2: Both husband and wife were screened by blood routine and hemoglobin electrophoresis. If one or both of them were positive, both would be tested for thalassemia gene. Mode 3: The couple received blood routine tests initially. If either was positive, both should receive hemoglobin electrophoresis testing. If either was positive, both parties will conduct thalassemia gene testing. Mode 4: The woman was screened by blood routine and hemoglobin electrophoresis. If any one of them was positive, the woman would be tested for thalassemia gene. If the gene test result was positive, the spouse should receive thalassemia gene. Mode 5: Both spouses conducted a blood routine test. If either was positive, both would conduct hemoglobin electrophoresis test. If both were positive, both spouses should receive thalassemia gene testing. Gene testing mode: The woman would be tested for thalassemia, and her spouse would have thalassemia test too if her result was positive.Results:When using MCV<80 fl as the cut-off for diagnosing thalassemia, the Youden indices of the five prenatal screening modes in Hunan Province were 0.551, 0.639, 0.898, 0.555 and 0.356, while when using MCV<82 fl as the cut-off, the Youden indices were 0.549, 0.629, 0.851, 0.548 and 0.356. When the MCV cut-off value was <80 fl, the missed diagnosis rates of the five screening modes were 44.44%, 0.00, 0.00, 18.52% and 62.96%, and the cost-effectiveness ratios were 21 709, 250 939, 76 870, 138 463 and 92 860 yuan (RMB)/couple, respectively. When the price of genetic testing was lower than 55 yuan (RMB), the cost-effectiveness ratio of genetic screening was lower than that of Mode 3.Conclusions:MCV<80 fl can be considered as the positive criteria in blood routine screening for thalassemia in Hunan Province, and the cost-effectiveness ratio of Mode 3 (the couple received blood routine tests initially. If either was positive, both should receive hemoglobin electrophoresis testing. If either was positive, both parties will conduct thalassemia gene testing) is the best. Genetic screening has certain advantages with the decreasing price.

AIM: To investigate the guiding role of individualized medication adjustment based on CYP2C19 metabolic typing in the treatment of ischemic stroke with clopidogrel, and to provide reference for clinical individualized medication. METHODS: The total of 80 patients with ischemic stroke were divided into the individualized drug instruction group with gene detection (n=40) and the control group without gene detection (n=40) according to whether they received CYP2C19 gene detection. According to the metabolism of CYP2C19, the individualized medication instruction group was divided into slow metabolic type, intermediate metabolic type, fast metabolic type and ultra-fast metabolic type. Patients with fast and ultra-fast metabolites were given clopidogrel dose of 75 mg once a day. Patients with intermediate metabolic type were given double clopidogrel dose of 150 mg once a day. Patients with slow metabolism were given tigrillo dose of 90 mg twice a day or aspirin dose of 100 mg once a day. The control group received 75 mg clopidogrel once a day. All patients enrolled in the groups were followed up for 3 months by outpatients or telephone. The incidence of vascular events and mRS scale scores were compared between the two groups. RESULTS: The incidence of vascular events in the individualized drug instruction group was significantly lower than that in the control group, and the incidence of mRS score(0-1) was significantly higher than that in the control group, with statistically significant differences (P<0.05). CONCLUSION: The individualized medication for patients with ischemic stroke by CYP2C19 gene detection can significantly reduce the incidence of adverse vascular events and improve the prognosis and living ability of patients.

OBJECTIVE: To investigate the protective effect of arctiin with anti-inflammatory bioactivity against triptolide-induced nephrotoxicity in rats and explore the underlying mechanism. METHODS: Forty SD rats were divided into 4 groups for gastric lavage of normal saline, arctiin (500 mg/kg), triptolide (500 μg/kg), or both arctiin (500 mg/kg) and triptolide (500 μg/kg). Blood samples were collected for analysis of biochemical renal parameters, and the renal tissues were harvested for determining the kidney index and for pathological evaluation with HE staining. In the RESULTS: In SD rats, arctiin significantly antagonized triptolide-induced elevation of BUN, Scr and kidney index ( CONCLUSIONS Arctiin can protect the kidney from triptolide-induced damages in rats possibly through the anti-inflammatory pathway.
Animals ; Anti-Inflammatory Agents ; Diterpenes/toxicity* ; Epoxy Compounds/toxicity* ; Furans ; Glucosides ; Kidney/drug effects* ; Phenanthrenes/toxicity* ; Rats ; Rats, Sprague-Dawley

Electroencephalogram (EEG) has been an important tool for scientists to study epilepsy and evaluate the treatment of epilepsy for half a century, since epilepsy seizures are caused by the diffusion of excessive discharge of brain neurons. This paper reviews the clinical application of scalp EEG in the treatment of intractable epilepsy with vagus nerve stimulation (VNS) in the past 30 years. It mainly introduces the prediction of the therapeutic effect of VNS on intractable epilepsy based on EEG characteristics and the effect of VNS on EEG of patients with intractable epilepsy, and expounds some therapeutic mechanisms of VNS. For predicting the efficacy of VNS based on EEG characteristics, EEG characteristics such as epileptiform discharge, polarity of slow cortical potential changes, changes of EEG symmetry level and changes of EEG power spectrum are described. In view of the influence of VNS treatment on patients' EEG characteristics, the change of epileptiform discharge, power spectrum, synchrony, brain network and amplitude of event-related potential P300 are described. Although no representative EEG markers have been identified for clinical promotion, this review paves the way for prospective studies of larger patient populations in the future to better apply EEG to the clinical treatment of VNS, and provides ideas for predicting VNS efficacy, assessing VNS efficacy, and understanding VNS treatment mechanisms, with broad medical and scientific implications.
Drug Resistant Epilepsy ; Electroencephalography ; Humans ; Prospective Studies ; Scalp ; Treatment Outcome ; Vagus Nerve Stimulation

Overexpression of exogenous lineage-determining factors succeeds in directly reprogramming fibroblasts to various cell types. Several studies have reported reprogramming of fibroblasts into induced cardiac progenitor cells (iCPCs). CRISPR/Cas9-mediated gene activation is a potential approach for cellular reprogramming due to its high precision and multiplexing capacity. Here we show lineage reprogramming to iCPCs through a dead Cas9 (dCas9)-based transcription activation system. Targeted and robust activation of endogenous cardiac factors, including GATA4, HAND2, MEF2C and TBX5 (G, H, M and T; GHMT), can reprogram human fibroblasts toward iCPCs. The iCPCs show potentials to differentiate into cardiomyocytes, smooth muscle cells and endothelial cells . Addition of MEIS1 to GHMT induces cell cycle arrest in G2/M and facilitates cardiac reprogramming. Lineage reprogramming of human fibroblasts into iCPCs provides a promising cellular resource for disease modeling, drug discovery and individualized cardiac cell therapy.

In the presence of silver ion, Mn2+ could be electro-oxidized to potassium hypermanganate in phosphoric acid solution, which could effectively react with pyrocatechol in acid solution and luminol in sodium hydroxide solution to produce chemiluminescence. On the basis of this, a novel indirect approach for the detection of Mn2+ was established. The effect of silver ions on the electrochemical oxidation of Mn2+was studied. when 1. 5 ×10-5 mol/L Ag+ and 0. 01 mol/L phosphoric acid solution were used in the process of electrochemical oxidation, the CL intensity could be up to the maximum value after the above solution was electrolyzed for 2 min. The relation of CL intensity and Mn2+concentration in the solutions at different pH and the selectivity were also investigated. when the pyrocatechol was used as luminescent reagent in the acidic medium, the CL intensity was linearly to the Mn2+concentration in the range of 1. 82×10-7-7. 27×10-5 mol/L with excellent selectivity. Common ions had little interferences in the determination of Mn2+. The method was successfully applied to the determination of Mn2+ in surface water and drinking water with satisfactory results.

Objective To observe the effects of Yishen Tongluo Decoction (YTD) on the renal mRNA expression of transforming growth factor-β1 ( TGF-β1) and collagen Ⅳ ( ColⅣ) in membranous nephropathy ( MN) rats. Methods SD rats were randomly divided into normal group, model group, benazepril group (in the dosage of 10 mg·kg-1·d-1) , YTD group ( in the dosage of 20 g·kg-1·d-1) . The rats in various groups were given intragastric administration of corresponding agents. At the end of the fourth week, 24-hour urinary protein quantity, albumin ( ALB) , total protein ( TP) , triglyceride ( TG) , total cholesterol ( TC) , blood urea nitrogen ( BUN) , and creatinine (Cr) levels were observed. The mRNA expression levels of TGF-β1 and ColⅣ in renal tissue of rats were detected by immunofluorescence method, electron microscopy and real-time polymerase chain reaction (PCR) method. Results In the model group, urinary protein quantity in rats was increased, serum levels of TP and ALB were significantly lowered, serum levels of TC and TG were significantly increased, renal pathological changes were present, and mRNA expression levels of TGF-β1 and ColIV in renal tissue were up-regulated (P<0.05 compared with normal group). Compared with the model group, 24-hour urinary protein quantity, TC and TG levels were significantly lowered, TP and ALB levels were significantly increased, rat renal injury was relieved, mRNA expression levels of TGF-β1 and ColIV in renal tissue were down-regulated in the treatment groups ( P<0.05) . However, the differences between benazepril group and YTD group were insignificant ( P>0.05) . Conclusion The therapeutic mechanism of YTD for MN is probably related with the inhibition of mRNA expression of TGF-β1 and ColⅣin renal tissue.

Objective To study the changes of serum high-sensitivity C-reactive protein (ha-CRP) and depression in patients with obstructive sleep apnea-hypopnea syndrome (OSAHS),and investigate the relationship between them.Methods Thirty healthy volunteers (control group) and 54 OSAHS patients (OSAHS group) were recruited for the study.The level of serum hs-CRP was determined by scatter rate nephelometry,and the state of depression was assessed by self-rating depression scale (SDS).Results The level of serum hs-CRP and SDS score were higher in OSAHS group than those in control group [(48.8 ± 12.7) scores vs.(36.3 ± 6.3) scores,(3.3 ±0.7) mg/L vs.(1.4 ± 0.4) mg/L](P＜0.01).SDS score and the level of serum hs-CRPwere positively correlated to apnea-hypopnea index(AHI) (r = 0.636,0.628 ;P<0.01) and negatively related to the MSaO2 (r =-0.509,-0.614;P <0.01) and LSaO2 (r =-0.607,-0.512;P <0.01).The level of serum hs-CRP was positive correlation to SDS score (r = 0.536,P<0.01).SDS score was related to the AHI,the level of serum hs-CRP and LSaO2 in multiple linear regression(F= 33.31,P = 0.002).Conclusion Depression is correlated to AHI and the level of serum hs-CRP in patients with OSAHS.

Objective To evaluate the effect and safety of tiotropium plus sustained-release theophylline in treating stable moderate chronic obstructive pulmonary diseases (COPD) . Methods A randomized clinical trail was conducted in 80 patients with stable moderate COPD, who were admitted to our hospital from March 2008 to December 2009. All patients were divided into two groups randomly and accepted tiotropium (groupⅠ ,n =40) or tiotropium plus sustained-release theophylline (group Ⅱ ,n =40) treatment for 12 weeks. Results Seventy patients completed the study,in which 36 cases had tiotropium alone and 34 cases had combination therapy. At the end of treatment, no statistically significant difference was found between the two groups in the comparisons of FEV1, FEV1%, FVC, FEV1/FVC, activity score and impacts score of the St George's Respiratory Questionnaire (SGRQ) (Ps > 0. 05). While the symptoms score (53.08 ± 12.25 vs.59.39±9.74,t=2.39,P<0.05) and total score (23. 91 ±2. 57 vs. 25. 27 ±2.14, t =2.40, P <0.05) of SGRQ were significantly lower in group Ⅱ compared to group Ⅰ. Six adverse events,including dry mouth (n =4), palpitation (re = 1) , constipation (n = 1), were observed in tiotropium alone group during the treatment period. Eleven adverse events were observed in combination treatment group, including dry mouth (re = 3) ,stomach discomfort or nausea (n = 3), diarrhea (re = 2) , palpitation (n = 2), constipation (n = 1) . The incidences of adverse reactions of two groups had no significant difference (P > 0. 05). Conclusion Compared with the single application of tiotropium, tiotropium plus sustained-release theophylline therapy can alleviate symptoms and improve life quality in patients with stable moderate COPD,without significantly increasing adverse reactions.