OBJECTIVE To optimize the extraction process of organic acids from Angelica sinensis using deep eutectic solvents （DESs）， and conduct characterization， antioxidant activity evaluation， and extraction mechanism analysis. METHODS The conductor-like screening model for realistic solvation with segment activity coefficients （COSMO-SAC） was employed to screen the types of DESs. With total organic acid content as the response value， single-factor experiments and Box-Behnken response-surface methodology were used to optimize the extraction conditions. Using A. sinensis decoction pieces and/or A. sinensis methanol extract as references， scanning electron microscope and Fourier transform infrared spectrometer （FTIR） were applied to characterize the products. Additionally， the 1，1-diphenyl-2-picrylhydrazyl （DPPH） and 2，2′-azino-bis（3-ethylbenzothiazoline-6- sulfonic acid） （ABTS） free radical scavenging capacities were determined. Density functional theory （DFT） was used to analyze the extraction mechanism of ferulic acid and chlorogenic acid by the DESs. RESULTS & CONCLUSIONS The optimal DESs was choline chloride-propanediol. The optimal extraction conditions for organic acids from A. sinensis were as follows： choline chloride- propanediol molar ratio of 1∶1， DESs water content of 70%， solid-liquid ratio of 1∶10， heating temperature of 57 ℃， and heating and stirring time of 8 min. In three validation experiments， the total organic acid content was 2.92 mg/g， yielding a relative error of 0.34% compared to the predicted value （2.91 mg/g）. Compared with A. sinensis decoction pieces and methanol extracts， the agglomerated structure of the DESs extract powder almost disappeared， showing the presence of lamellar structures similar to those of the intestinal wall. Compared with the methanol extract， the DES extract exhibited higher FTIR characteristic peak intensity and peak area integration， as well as stronger scavenging capacities against DPPH and ABTS free radicals. The extraction of organic acids from A. sinensis by DESs is the result of the combined effects of polarity matching， hydrogen bonding， and structural adaptation.

Objective To explore the effect of jogging on the patients with depression disorder of liver-stagnation and spleen-deficiency.Methods 100 patients with liver-stagnation and spleen-deficiency depression disorder were randomly selected from Tongde Hospital of Zhejiang Province from March to September 2022.They were divided into observation group and control group(50 cases each)by random number table method.The control group received conventional antidepressant drug treatment,and the observation group was given jogging exercise regimen on the basis of the control group:exercise at least 4 times a week,30-60 min each time,and the heart rate was maintained at 110-120 times/min for 4 weeks.The depressive disorder of the patients was assessed by Zung's Self-rating Depression Scale(SDS)and 17-item Hamilton Depression Scale(HAMD-17).Results After jogging exercise intervention,the total effective rate of the observation group was 94%,which was significantly higher than that of the control group 80%(P<0.05).HAMD-17 score was significantly lower in the observation group than in the control group after intervention(F=12.575,P<0.05).The scores of cognitive impairment,block and sleep disturbance in the observation group were significantly lower than those in the control group after intervention(F=12.575,P<0.05).The body weight factor scores of the observation group and the control group were significantly higher after intervention than before intervention(P>0.05).Conclusions Jogging exercise intervention therapy can significantly improve the depression of patients with liver-qi and spleen-deficiency depression disorder,and jogging can be an effective method to prevent and treat depression disorder.

The clinical data, laboratory test, and gene mutations were collected from a family with pseudohypoaldosteronism type II(PHA2). The proband, aged 1 year and 7 months, presented with hyperkalemia(6.69 mmol/L; reference range 3.5-5.3 mmol/L), blood pressure of 110/68 mmHg(normal<106/61 mmHg, 1 mmHg=0.133 kPa), blood chloride of 111.5 mmol/L(reference 99-110 mmol/L), blood HCO 3- of 17.1 mmol/L(reference 22-29 mmol/L), estimated glomerular filtration rate of 128.5 mL·min -1·(1.73 m 2) -1[>90 mL·min -1·(1.73 m 2) -1], and blood renin concentration of 0.30 μIU/mL(reference 4.2-45.6 μIU/mL). The mother and maternal grandfather also exhibited normal renal function with hyperkalemia, hypertension, hyperchloremia, metabolic acidosis, and low renin. Genetic testing revealed a heterozygous missense mutation(c.1685A>G, p. E562G) in exon 7 of the no-lysine kinase 4(WNK4) gene. Treatment with hydrochlorothiazide was effective. Literature review comparing this E562G pedigree with other WNK4 variants suggested clinical heterogeneity of WNK4 mutations. For unexplained hyperkalemia, especially with concurrent hypertension, PHA2 should be considered early for genetic screening to prevent misdiagnosis or delayed diagnosis.

Objective:To investigate the relationship between the levels of serum cytokines and chemokines and the prognosis of patients with acute B-ALL after receiving chimeric antigen receptor (CAR)-T cell immunotherapy and acute graft-versus-host disease (aGVHD) in patients after bridging allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:According to the case-control principle, Forty-two patients with B-ALL who received CD19-CAR-T cell immunotherapy bridged to allo-HSCT at Heibei Yanda Ludaopei Hospital from September 18, 2019 to May 9, 2022 were enrolled. Mann-Whitney U test was used to compare the changes of aGVHD-related cytokines and chemokine levels between CAR-T cell immunotherapy and bridging transplantation in different patients at the same time. Their plasma levels of cytokines and chemokines related to aGVHD were monitored at the day before CAR-T therapy and after CAR-T treatment at day 4, 7,14,21,28. The receiver operating characteristic curve was drawn to evaluate the predictive value of cytokines and chemokines in predicting the occurrence and the death of aGVHD patients. Kaplan-Meier method and Log-rank tests were used for Overall survival (OS) analysis. Results:Twenty-four of total 42 patients had aGVHD, of which 11 patients died and 31 patients survived. There was no significant difference in cytokines and chemokines between the aGVHD group and the non-aGVHD group on the day before CAR-T cell treatment. According to statistical analysis, the serum Elafin levels of aGVHD group was higher than that of non-aGVHD group at the 21st day [4 482 (2 811, 6 061) ng/L vs 2 466 (1 948, 3 375) ng/L, Z=3.145, P=0.001] and the 28st day [4 391 (2 808, 5594) ng/L vs 2 463 (1 658, 2 830) ng/L, Z=2.038, P=0.048] separately. At the 14th day, serum cytokines and chemokines levels between the two group were as follows,MIP-1 α [21.02 (12.36, 30.35) ng/L vs 5.56 (3.64, 10.79) ng/L], sCD25 [422.47 (257.99, 1 233.78) IU/ml vs 216.11 (133.75,457.39) IU/ml], Elafin [4 101 (2 393, 5 006) ng/L vs 2 155 (1 781, 3 033) ng/L], IL-6 [119.08 (23.97, 183.43) ng/L vs 8.39 (2.91, 17.42) ng/L] and IL-8 [13.56 (12.50, 24.52) ng/L vs 2.83 (1.73,6.87) ng/L] were at higher levels ( Z=2.653, P=0.007; Z=2.176, P=0. 030; Z=2.058, P=0.041; Z=3.329, P<0.001; Z=3.162, P=0.001). The KM survival curve showed that the cumulative survival rates of patients with higher serum levels of MIP-1α, sCD25, Elafin, IL-6 and IL-8 were lower than those with low levels at day 14, and the difference was statistically significant (χ 2=12.353, 4.890, 6.551, 10.563, 20.755, P<0.05). Conclusion:The outcomes of patients treated with CAR-T cell therapy bridged to allo-HSCT was correlated with serum MIP-1α, sCD25, Elafin, IL-6 and IL-8 levels after receiving CAR-T therapy. High concentrations of MIP-1α, sCD25, Elafin, IL-6 and IL-8 suggest poor prognosis and can be used as biomarkers to suggest appropriate clinical selection of therapy.

Objective: To investigate the effects of estrogen deficiency on depression-like behavior and mitochondrial function in hippocampus，and to study the underlying mechanisms． Methods: The female adult C57BL /6 mice were randomly assigned to Sham group，Ovx group and Ovx + E2 group．On the 7th day after operation，Ovx + E2 group was subcutaneously injected with 17 β-estradiol ( E2 ) for 4 weeks．Then another 48 female adult C57BL /6 mice were randomly divided into Sham group，Sham + MnTBAP group，Ovx group and Ovx + MnTBAP group．After operation for one week，MnTBAP was injected into the hippocampus of mice in Sham + MnTBAP group and Ovx + MnTBAP group．The depression-like behaviors of the mice were monitored by sucrose preference test and forced swimming test．Then the mice were decapitated，and the hippocampus samples were collected．Mitochondria of hippocampus were isolated using mitochondria fractionation kit．The concentrations of adenosine triphosphate (ATP) ， reactive oxygen species ( ROS) and mitochondrial membrane potential were determined，respectively. Results: Compared with Sham mice，Ovx showed a decrease in the percentage of sucrose consumption，an increase in immobility time and the alterations of hippocampal mitochondrial function，which were reversed by estrogen treatment. Intrahippocampal injection of MnTBAP significantly increased the percentage of sucrose consumption and decreased immobility time in Ovx mice． Conclusion Hippocampal mitochondrial dysfunction contributes to estrogen deficiency-induced depression-like behaviors.

Objective To analyze the clinical characteristics and genotype of a patient with congenital systemic lip-odystrophy(CGL)type 1 associated with exudative xanthoma caused by AGPAT2 gene mutation,and to provide ev-idence for clinical and genetic diagnosis of the disease.Methods Clinical data of the patient such as medical histo-ry,physical examination and laboratory examination were collected.Peripheral venous blood was collected for whole exome sequencing analysis and Sanger sequencing verification,and treatment was provided to patients according to the changes of condition.Results The clinical manifestations of the patient were subcutaneous fat reduction,fatty liver,spleen enlargement,kidney enlargement,high blood sugar and lipids,severe insulin resistance,scattered yellow rash on limbs,which was confirmed as xanthoma.The results of whole exon sequencing showed that the AGPAT2 gene of the patient had a heterozygous nonsense mutation of c.202C＞T:p.R68?and c.646A＞T:p.K216?,and the former was the pathogenic mutation site.Follow-up therapy covers improvement of lifestyle,low-fat diet and regular exercise.The rashes subsided after active lipid-lowering therapy.Conclusions Apart from typical lipody-strophy,the patient was accompanied by exanthemous xanthoma.No CGL1 patient with exanthemous xanthoma has been reported in the domestic literature database up to now,and the genetic test results showed that there was a c.202C＞T heterozygous mutation of AGPAT2 gene.This gene site has not been reported in the literature,and its functional verification needs to be further studied.

Objective:To investigate the prognostic value of colony forming unit-granulocyte and macrophage (CFU-GM) in allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:Seventy-three patients who received allo-HSCT in Hebei Yanda Lu Daopei Hospital from February 2015 to January 2017 were selected. According to the level of CFU-GM from bone marrow (BM) culture at the time of allo-HSCT, the patients were fit into high CFU-GM group and low CFU-GM group. The overall survival rate (OS) and relapse-free mortality rate (NRM) of patients after transplantation were tested by χ2 test after a follow-up of 37.0 (12.5, 50.5) months. Kaplan-Meier method was used to compare OS and event-free survival (EFS) of patients with different CFU-GM levels. Logistic regression model was used to analyze the prognostic factors. Cox regression model was used to further analyze the prognostic risk of patients.Results:Compared with the low CFU-GM group, the high CFU-GM group had a higher OS (81.40% vs 60.00%, χ2=4.067, P=0.044) and a lower NRM (11.63% vs 36.67%, χ2=6.474, P=0.011). Compared with the low CFU-GM group, the mean OS time (57.6 and 37.1 months, respectively, P=0.039) and the mean EFS time (61.7 and 38.5 months, respectively, P=0.011) were significantly higher in the high CFU-GM group. Logistic regression analysis showed that both the level of CFU-GM and BM MNC were significant influencing factors of OS ( OR=2.917, 95% CI 1.011-8.418, P=0.048 and OR=1.510, 95% CI 1.058-2.154, P=0.023, respectively) and EFS ( OR=4.400, 95% CI 1.336-14.492, P=0.015 and OR=1.447, 95% CI 1.002-2.090, P=0.049, respectively)after transplantation. The level of CFU-GM was an independent risk factor for evaluating EFS ( HR=0.279, 95% CI 0.097-0.805, P=0.018). BM MNC was an independent risk factor for OS ( HR=1.345, 95% CI 1.052-1.720, P=0.018). Conclusion:The level of CFU-GM and BM MNC were related to the prognosis of allo-HSCT. The patients in the high CFU-GM group had higher EFS.

Objective:To investigate the effect of SCN1A gene polymorphism (SCN1A-rs3812718) on the alterations of spontaneous brain activity using amplitude of low-frequency fluctuations (ALFF) of MR in patients with temporal lobe epilepsy (TLE).Methods:A total of 37 TLE patients (TLE group) admitted to the Epilepsy Center of the 900th Hospital of Joint Logistic Team from March 2018 to August 2019 were retrospectively analyzed, and another 28 healthy volunteers matched for gender, age, and years of education with the TLE group were selected as the healthy control group (HC group). Sixty-five subjects were divided into four groups by genotype and diagnosis: 34 cases in AA/AG-TLE subgroup, 3 cases in GG-TLE subgroup, 20 cases in AA/AG-HC subgroup and 8 cases in GG-HC subgroup. All subjects underwent sagittal 3D-T 1WI and resting-state functional MRI using a Siemens 3.0 T Trio Tim MR scanner. Then ALFF values of the four groups were calculated using DPABI by the MATLAB 2010 platform. The ALFF values between two groups were compared by independent samples t-test. The ALFF values of different genotypes at rs3812718 locus in TLE and HC group were analyzed by multivariate analysis of variance to find out the corresponding brain regions with interaction, and then post hoc simple effect analysis was performed. Results:The ALFF values in TLE group significantly increased in left marginal lobe, left parahippocampal gyrus, left fusiform gyrus, left hippocampus, right insular lobe and right inferior temporal gyrus (Alphasim corrected P<0.001) and decreased in the left superior frontal gyrus, left middle frontal gyrus, left inferior frontal gyrus, right middle frontal gyrus, right precuneus, left precuneus, bilateral cingulate gyrus and right angular gyrus (Alphasim correction P<0.05) compared with HC group. Subjects carrying the non-risk G allele had higher ALFF values in the right inferior temporal gyrus, right fusiform gyrus, and right cerebellum than subjects carrying the risk A allele ( t=3.30, Alphasim corrected P=0.002). There was a significant interaction effect on posterior cerebellar lobe, left anterior cerebellar lobe, left inferior temporal gyrus, left superior parietal lobule and right precuneus of TLE patients with SCN1A-rs3812718 genotype. Post-hoc simple effect analysis showed that ALFF significantly increased in the left posterior cerebellar lobe, left anterior cerebellar lobe, left inferior temporal gyrus and left fusiform gyrus in GG-TLE subgroup ( t=5.97, P<0.001), but significantly decreased in the right superior parietal lobule, right precuneus, right posterior cerebellar lobe in AA/AG-TLE subgroup compared to the HC group. Compared with GG-TLE subgroup, ALFF in left posterior cerebellar lobe, left fusiform gyrus and left inferior temporal gyrus decreased in AA/AG-TLE subgroup. Conclusion:SCN1A gene polymorphism in the rs3812718 locus affects spontaneous neural activity in resting state, which may be one of the pathophysiological mechanisms of TLE.

Objective:To investigate the significance of multicolor flow cytometry (MFC) monitoring of minimal residual disease (MRD) in the course of allogeneic hematopoietic stem cell transplantation (allo-HSCT) after CD19-chimeric antigen receptor(CAR)-T cell immunotherapy for patients with refractory, relapsed B-cell acute lymphoblastic leukemia (r/r B-ALL).Methods:37 patients with r/r B-ALL admitted to Hebei Yanda Lu Daopei Hospital from January to July 2019, aged 15 (6, 19) years old, including 24 males and 13 females, were treated with CD19-CAR-T cell immunotherapy bridging allo-HSCT. MFC with cytoplasmic CD79a antibody to set up B-cell gates was used to monitor patients′ bone marrow (BM), cerebrospinal fluid (CSF), and tissue samples on day 0 (prior to the CAR-T cell immunotherapy), day 15, day 28 post CAR-T cell immunotherapy, and post transplantation.The MRD values of these samples were analyzed to evaluate the residual tumor cells and metastasis. The killing effect of the CAR-T cells was evaluated by the recovery of CD19+B cells before transplantation and the period between the timepoint when CD19+B cells was recovered and the timepoint when CAR-T cells were infused. Peripheral blood CAR-T cells were counted at different time points. Statistic analysis was performed by Kaplan-Meie assay and Log-rank test to analyze the difference of univariate cumulative survival.Results:(1)Among the 37 patients, 8 died and 29 survived. 5 patients relapsed after transplantation, of which 4 relapsed patients died and 1 survived. (2)MFC MRD negative remission rate of the death group was lower than that of the survival group at the following time points: post-CAR-T therapy and prior to transplantation (5/8 vs. 28/29, χ 2=7.540, P=0.006); day 15 of the CAR-T cell reinfusion (3/8 vs. 24/29, χ 2=6.512, P=0.011); day 28 of the reinfusion (3/8 vs. 276/29, χ 2=10.065, P=0.002). The probability of extramedullary MFC MRD positive tumor infiltration in the death group was higher than that in the survival group(7/8 vs. 14/29, χ 2=3.931, P=0.047). After CAR-T cell immunotherapy, the recovery period of CD19-positive cells in the death group, or the time for CAR-T cells to kill CD19-positive cells, was shorter than that in the survival group [42.00 days(30.00,49.00) vs. 55.00 days(41.50,73.50), Z=0.022, P=0.020]. Conclusion:The positive results of MRD by MFC at the following timepoints may predict unfavorable outcomes, such as post-CAR-T therapy and prior to transplantation, day 15 and 28 of the CAR-T cell immunotherapy, which may provide some guidance for clinical management.

Objective:To design and validate a high-quality rapid screening questionnaire based on the common medical history and clinical experience of benign paroxysmal positional vertigo (BPPV).Methods:A questionnaire was designed based on expert′s opinions, and the first-time patients who complained of dizziness and vertigo in the vertigo clinic of the First Hospital of Shanxi Medical University from September 2020 to June 2021 were prospectively screened. Taking the displacement test as the gold standard, the reliability and validity of the questionnaire were tested to evaluate its authenticity, reliability and benefit value. This study was divided into three steps. The first step was to conduct a pre-experiment and to adjust the questionnaire items; the second step was to determine the questionnaire items and the best cut-off value; the third step was to screen patients with the best cut-off value and to evaluate the quality of the questionnaire.Results:Seven items were finalized. The Cronbach′s coefficient of the questionnaire was 0.675, the content validity was 0.85, the KMO value of the construct validity was 0.648, and there were 4 factors with characteristic root>1, and the cumulative contribution rate was 76.309%. The area under the receiver operating characteristic curve (ROC) was 0.938, and its optimal cut-off value was 4.5 points. At this point, the sensitivity was 88.89% and the specificity was 85.44%.Conclusion:The BPPV rapid screening questionnaire has high sensitivity and specificity, which can be used for clinical screening of BPPV patients.