Objective:To construct dual-targeting(CD38 and CD138)chimeric antigen receptor(CAR)gene-modified T cells(CD38/CD138 CAR-T cells)and explore their in vitro cytotoxicity against multiple myeloma(MM)cells.Methods:Based on the high expression of CD38 and CD138 antigens in MM cells,CD38 CAR-T cells and CD138 CAR-T cells targeting CD38 and CD138 respectively,and CD38/CD138 dual-targeted CAR-T cells targeting both CD38 and CD138 were constructed using CAR-T cell technology.The experimental groups included untreated T cells,CD38 CAR-T,CD138 CAR-T,and CD38/CD138 CAR-T cells.The phenotype of CAR-T cells was detected by flow cytometry.The cytotoxicity of various CAR-T cells against MM cells(RPMI8226 and U266)was assessed using the LDH release assay.Results:Three types of CAR-T cells,CD38 CAR-T,CD138 CAR-T,and CD38/CD138 CAR-T cells,were successfully constructed.The CD38/CD138 CAR-T cells tended to differentiate into a memory phenotype,expressing higher levels of proliferation marker(CD25),activation marker(CD27),and lower levels of exhaustion markers(PD-1,CTLA-4,TIM-3)(all P<0.001).Moreover,CD38/CD138 CAR-T cells were less prone to exhaustion and senescence,and expressed lower levels of r-H2AX,p-p53,p21,and p16 proteins(all P<0.01).Under different effector-to-target cell ratios,CD38/CD138 CAR-T cells exhibited stronger cytotoxic effects against RPMI8226 and U266 cells compared to CD38 CAR-T and CD138 CAR-T cells(all P<0.001).Conclusion:CD38/CD138 CAR-T cells targeting both CD38 and CD138 demonstrate an optimal phenotype and enhanced anti-tumor activity in vitro,offering promising potential for immunotherapy in multiple myeloma.

Objective To investigate the effect and underlying mechanism of SULT2B1 in the de-velopment of atherosclerosis(AS).Methods Twelve 8-week-old apolipoprotein E-knockout(apoE-/-)male mice were subjected and fed with a high-fat diet for 12 weeks,and then randomly divided into adeno-associated virus(AAV)-GFP and AAV-shSULT2B1 groups,with 6 animals in each group.In 4 weeks after AAV injection via tail vein,the mice were sacrificed for assessing aortic and aortic root plaque formation by oil red O staining and detecting serum levels of inflam-matory factors and blood lipids.RAW264.7 cells were transfected with adenovirus(Ad)-GFP and Ad-SULT2B1,respectively(n=3).RNA sequencing was performed to detect downstream RNA changes.Then LncRNA gga3-204 was selected for downstream study.After RAW264.7 cells were divided into si-NC group,si-SULT2B1 group,si-Lncgga3-204 group and si-SULT2B1+si-Lncg-ga3-204 group(n=3),and the IL-1β and IL-6 levels were detected in these transfected cells.Results There was no statistically difference in body weight in the mice from the AAV-GFP and AAV-shSULT2B1 groups after high-fat feeding(P＞0.05).Significantly lower serum levels of TC,TG and LDL-C,reduced aortic plaque area[(8.38±1.33)％vs(11.83±1.04)％,P=0.000],and decreased TG content within the aortic root plaque[(12.29±1.54)％vs(17.67±1.53)％,P=0.000]were observed in the AAV-shSULT2B1 group than those in the AAV-GFP group.Ser-um IL-1β and IL-6 levels in the mice of the AAV-shSULT2B1 group than those in the AAV-GFP group(P＜0.01).The AAV-shSULT2B1 group also had obviously lower serum levels of I L-1 βand IL-6 than the AAV-GFP group(P＜0.01).In the RAW264.7 cells from the si-SULT2B1 group,the mRNA levels of IL-1β and IL-6 were notably lower than those in the si-NC group(P＜0.01).LncRNA gga3-204 expression was significantly higher in the Ad-shSULT2B1 group than the Ad-GFP group(P＜0.01).While,the si-SULT2B1 group had statistically higher Lncgga3-204 level than the si-NC group(2.32±0.60 vs 1.19±0.21,P=0.036).The si-Lncgga3-204 group had significantly higher IL-1β and IL-6 mRNA levels than the si-NC group(P＜0.01).The si-SULT2B1+si-Lncgga3-204 group had significantly higher IL-1β and IL-6 mRNA levels than the si-SULT2B1 group(P＜0.05).Conclusion SULT2B1 affects the macrophage inflammatory response via Lncgga3-204,and then affects the progression of AS.

Objective:To summarize clinical experience in the management of Abernethy malformation combined with focal nodular hyperplasia (FNH).Methods:Three patients with Abernethy malformation combined with FNH who were managed in the Department of Hepatobiliary Surgery, the First Medical Center of Chinese PLA General Hospital from January 2010 to December 2018 entered into this study. There were one male aged 35 years and two females aged 21 years and three years and nine months, respectively. Patients underwent digital subtraction angiography (DSA) with balloon dilation, shunt vasulature ligation, portal vein (PV) pressure measurement and other investigations were performed to study the changes of PV pressure before and after treatment.Results:DSA indicated that the congenital portal shunt of the patients was Abernethy deformity Ⅱ and Ⅰb, respectively, and with combined FNH diagnosed preoperatively. One patient was admitted to hospital for partial hepatectomy 5 years ago for the treatment of FNH, which presented with recurrent abdominal pain. The PV pressure increased from 8.5, 9.0, 20.0 cmH 2O(1 cmH 2O=0.098 kPa) to 15.0, 21.0, 25.0 cmH 2O after portal cavity shunt vasculature/portal vein ligation and splenic artery ligation/portal body shunt ligation, respectively. The PV blood flow into the liver increased, and DSA suggested shunt vessel occlusion. On follow-up, all patients survived well. Conclusion:Ligation or constriction of shunt vessels significantly improved the flow of PV into liver in Abernethy malformation patients with FNH. The treatment benefited long-term survival.

Objective:To investigate the value of neutrophil-to-lymphocyte ratio (NLR) in the diagnosis and prognosis evaluation of primary hepatocellular carcinoma.Methods:The clinical data of 100 patients pathologically diagnosed as primary liver cancer who were admitted to the First Medical Center of Chinese PLA General Hospital from January 2013 to December 2015 were retrospectively analyzed. Serum alpha fetoprotein (AFP), a conventional marker for hepatocellular carcinoma diagnosis was used as the control. The fourfold table diagnostic test was applied to analyze the sensitivity and specificity of serum NLR in the diagnosis of hepatocellular carcinoma, and the correlation with the degree of tumor differentiation was also analyzed.Results:The proportion of patients with high NLR (≥1.70) [56% (56/100)] was higher than the proportion of patients with positive AFP [44% (44/100)] in all 100 hepatocellular carcinoma patients, but the difference was not statistically significant ( χ2 = 2.88, P = 0.08). Among AFP-positive patients, the median survival time of patients with low and high NLR was 59 and 48 months, respectively, and the difference was statistically significant ( χ2 = 3.91, P = 0.048), and high NLR was an independent risk factor affecting the prognosis of hepatocellular carcinoma patients ( HR = 1.232, 95% CI 1.055-1.438, P = 0.008). Conclusions:The detection of NLR combined with AFP can improve the diagnostic rate of hepatocellular carcinoma before surgery. High NLR is an independent risk factor affecting the prognosis of patients with primary hepatocellular carcinoma.