Background and purpose:Chimeric antigen receptor T(CAR-T)cell therapy has shown remarkable efficacy in treating hematological and lymphatic system tumors,but its effectiveness in solid tumors is relatively poor,which is partly attributed to target selection.For Ewing sarcoma(ES),CD99 can be a potential target for CAR-T cells.However,due to T cells'endogenous expression of CD99 protein,CAR-T cells targeting CD99 face limitations in their expansion capacity in vitro.This study aimed to identify the optimal conditions for preparing CD99 CAR-T cells by incorporating CD99 knockdown short hairpin RNA(shRNA),optimizing the multiplicity of infection(MOI)for lentiviral transduction,and screening for the best culture medium and container for CAR-T cell expansion.Methods:shRNA sequences were screened to enhance the expansion capacity of CD99 CAR-T cells.Different MOI,culture media,and containers were used to assess CAR-T cell transduction efficiency,cell viability,proliferation capacity,specific killing ability,and interferon-γ(IFN-γ)release levels under various conditions,in order to identify the optimal cell preparation conditions.Results:The expansion level of KO-CD99 CAR-T cells obtained through shRNA knockdown was significantly higher than that of CD99 CAR-T cells[(16.40±0.40)vs(6.33±1.53),P<0.01].The optimal expansion effect was observed when the transduction MOI was between 0.25 and 1.0,and OptiVitro was used as the culture medium.CAR-T cells cultured in ventilated flasks exhibited significantly higher expansion rates compared to cells cultured in bags[MOI=0.25:(50.23±3.32)vs(13.02±4.82);MOI=0.50:(49.96±0.83)vs(18.25±2.88);MOI=1.00:(48.27±5.08)vs(13.16±6.26);P<0.01],with better cell phenotype and higher specific killing ability.Conclusion:KO-CD99 CAR-T cells obtained through shRNA technology can achieve stable expansion.Based on the optimization of expansion conditions,KO-CD99 CAR-T cells exhibit superior expansion capacity and a higher proportion of memory T cells when the MOI is between 0.25 and 1.00,OptiVitro is used as the culture medium,and ventilated flasks are used as the culture container.These findings lay a solid foundation for further clinical trials of CD99 CAR-T cell therapy for ES.

Objectives:To examine the efficacy of terminal branches portal vein embolization(TBPVE) for the increment of FLR in hepatocellular carcinoma (HCC) patients and to introduce its clinical value with transcatheter chemoembolization(TACE) in the treatment of HCC patients without surgery.Methods:One hundred and fifty HCC patients from three clinical centers of china underwent TBPVE technique from December 2016 to May 2021,including 89 males and 61 females. The average age was 51.9 years(range:18 to 79 years).One hundred and one patients were diagnosed with a background of HBV infection,including 27 patients with portal venous hypertension.TACE was performed simultaneously with TBPVE in 102 patients.Fifty-three patients underwent hepatectomy,who were subdivided into HBV positive and HBV negative groups,with TACE and without TACE groups to analyze the increment of future liver remnant (FLR), complications and survival data.These data were also analyzed in other 97 patients without hepatectomy.Results:All the patients reached adequate FLR successfully in 14 days after TBPVE including patients with portal venous hypertension.The average increment rates of FLR was 56.2% in 7 days and 57.8% in 14 days after TBPVE. There was no significant difference neither between HBV positive and HBV negative groups(7 days:(55.0±27.3)% vs.(57.8±20.9)%, t=0.885, P=0.373; 14 days:(57.3±24.6)% vs.(58.3±23.7)%; t=0.801, P=0.447),or between with TACE and without TACE groups(7 days:(62.3±26.3)% vs. (48.8±20.6)%; t=1.788, P=0.077;14 days:(64.4±25.0)% vs.(55.2±23.1)%; t=1.097, P=0.257).The morbidity and mortality rates were 20.8% and 1.9% in patients with hepatectomy.The 1-,3-year overall survival(OS) and disease-free(DFS) rates were 87.5%,64.5% and 64.7%,40.6% for patients underwent surgery.There was no significant difference of 1-,3-year OS and DFS between HBV positive and negative groups,but there were different between TACE and without TACE groups.The 1-,3-year OS for patients underwent TBPVE and TACE but without surgery were 80.1%, 53.7%. Conclusion:TBPVE is a good alternative technique for modulation of FLR for staged hepatectomy even in HBV positive HCC patients and can be applied with TACE procedure simultaneously as an option treatment for patients with no intend to surgery.

Objectives:To examine the efficacy of terminal branches portal vein embolization(TBPVE) for the increment of FLR in hepatocellular carcinoma (HCC) patients and to introduce its clinical value with transcatheter chemoembolization(TACE) in the treatment of HCC patients without surgery.Methods:One hundred and fifty HCC patients from three clinical centers of china underwent TBPVE technique from December 2016 to May 2021,including 89 males and 61 females. The average age was 51.9 years(range:18 to 79 years).One hundred and one patients were diagnosed with a background of HBV infection,including 27 patients with portal venous hypertension.TACE was performed simultaneously with TBPVE in 102 patients.Fifty-three patients underwent hepatectomy,who were subdivided into HBV positive and HBV negative groups,with TACE and without TACE groups to analyze the increment of future liver remnant (FLR), complications and survival data.These data were also analyzed in other 97 patients without hepatectomy.Results:All the patients reached adequate FLR successfully in 14 days after TBPVE including patients with portal venous hypertension.The average increment rates of FLR was 56.2% in 7 days and 57.8% in 14 days after TBPVE. There was no significant difference neither between HBV positive and HBV negative groups(7 days:(55.0±27.3)% vs.(57.8±20.9)%, t=0.885, P=0.373; 14 days:(57.3±24.6)% vs.(58.3±23.7)%; t=0.801, P=0.447),or between with TACE and without TACE groups(7 days:(62.3±26.3)% vs. (48.8±20.6)%; t=1.788, P=0.077;14 days:(64.4±25.0)% vs.(55.2±23.1)%; t=1.097, P=0.257).The morbidity and mortality rates were 20.8% and 1.9% in patients with hepatectomy.The 1-,3-year overall survival(OS) and disease-free(DFS) rates were 87.5%,64.5% and 64.7%,40.6% for patients underwent surgery.There was no significant difference of 1-,3-year OS and DFS between HBV positive and negative groups,but there were different between TACE and without TACE groups.The 1-,3-year OS for patients underwent TBPVE and TACE but without surgery were 80.1%, 53.7%. Conclusion:TBPVE is a good alternative technique for modulation of FLR for staged hepatectomy even in HBV positive HCC patients and can be applied with TACE procedure simultaneously as an option treatment for patients with no intend to surgery.

Objective:To investigate the correlation between subclinical hypothyroidism(SCH)and the degree of coronary artery stenosis in patients with coronary heart disease.Methods:From March 2018 to September 2019, 138 patients undergoing coronary angiography in our hospital were randomly selected and their serum thyroxine(FT4)and high-sensitivity thyroid stimulating hormone(sTSH)levels were measured.Based on the levels of FT4 and sTSH, patients were divided into the SCH group and the normal group.Combined with coronary angiography results, the correlation between the SCH and the number of coronary lesions were analyzed.Results:Patients in the SCH group were associated with significantly higher levels of cholesterol(TC)[(5.83±1.27)mmol/L vs.(5.02±1.22)mmol/L, t=3.746, P=0.000], triglyceride(TG)[(3.29±1.74)mmol/L vs.(2.17±1.68)mmol/L, t=3.769, P=0.000], low-density lipoprotein cholesterol(LDL-C)[(3.81±1.02)mmol/L vs.(3.24±1.08)mmol/L, t=3.092, P=0.001], and high-density lipoprotein cholesterol(HDL-C)[(1.13±0.27)mmol/L vs.(1.02±0.25)mmol/L, t=2.4459, P=0.008]than those in the normal group.There were significant differences in sTSH levels between patients with multivessel, double vessel or single vessel disease when grouped by the number of coronary lesions, and between those with severe, moderate or mild coronary artery stenosis when grouped by disease severity(all P<0.05). Multiple regression analysis showed that SCH, TC, and LDL-C were factors affecting the number of coronary lesions( P<0.05). Conclusions:SCH is an important factor that causes the occurrence and progression of coronary artery stenosis in patients with coronary heart disease, mainly by affecting lipid metabolism.

Objective To investigate the efficacy and safety of percutanous transhepatic intrahepatic portosystemic shunt(PTIPS)for chronic portal vein occlusion and cavernous transformation with symptomatic portal hypertension.Methods The clinical and imaging data of 38 patients with chronic portal vein occlusion and cavernous transformation with symptomatic portal hypertension, who received PTIPS in our hospital from November 2009 to June 2016,were analyzed retrospectively.The differences of the portosystemic pressure gradient(PPG)measured before and after PTIPS procedure was analyzed by a paired samples t-test. All the patients were followed up and the curative effect and operation-correlated complications were observed.Results The PTIPS procedure was technically successful in 36 patients.The other two patients with unsuccessful PTIPS underwent medical treatment,and one of them died of recurrent variceal bleeding 25 months later. Effective portal decompression and free antegrade shunt flow were achieved in 36 patients with successful PTIPS.And the mean PPG was decreased from(25.2±2.9)to(13.2± 1.3) mmHg (1 mmHg=0.133 kPa) before and after PTIPS respectively and the difference was statistically significant(P<0.05).During the procedure,arterial hemorrhage occurred in two patients who subsequently underwent embolization. Biliary injury occurred in one case and percutanous transhepatic biliary drainage (PTBD)was then performed.The mean follow-up period of the 36 patients was(26.7±10.4)months(range from 3.0 to 74.0 months).Hepatic encephalopathy appeared in 4 cases,among which,3 patients recovered after receiving medical treatment, while 1 patient experienced Grade 3 hepatic encephalopathy and recovered after implanting a smaller cover-stent.Shunt dysfunction occurred in 10 cases,of which 8 cases recovered after shunt revision with stent implantation or ballon angioplasty, while 2 cases underwent anticoagulation by warfarin only. During follow-up period, 7 patients died of liver failure(n=4), hepatic cellular carcinoma(n=1), recurrent varicose vein bleeding(n=1), and renal failure(n=1). The other patients remained asymptomatic and shunt patency. Conclusions PTIPS is both safe and effective for the treatment of symptomatic portal hypertension caused by chronic portal vein occlusion and cavernous transformation.The technical success rate is high,and the short-term curative effect is satisfied.

Objective To compare the anesthetic effect of ropivacaine and bupivacaine in combined spinal-epidural anesthesia(CSEA)for cesarean section,and their influence on the incidence rate of supine hypotension syndrome(SHS).Methods 200 patients with cesarean section surgery in our hospital from February 2016 to July 2016 were randomly divided into observation group and control group,all of cases were given CSEA.100 patients in the observation group(the group L)were given ropivacaine in spinal anesthesia,the other 100 patients in the control group(the group B)were given bupivacane in spinal anesthesia.Recorded the relevant indicators,compared the incidence rate of SHS,the effect of anesthesia and neonatal score.Results The incidence rate of SHS of the group L was lower than the group B(χ2 =9.261,P<0.01).The effect of anesthesia and Apgar score of two groups had no statistically significant differences(all P>0.05).Conclusion The application of ropivacaine in CSEA for cesarean section not only has exact anesthesia effect,but also can effectively prevent SHS without any side effects.

Objective: To analyze the efficacy of branches portal vein embolization (TBPVE) combined with transcatheter arterial chemoembolization (TACE) on liver neoplasms. Methods: From August 2016 to May 2017, there were 13 patients including 11 males and 2 females with primary hepatocellular carcinoma who underwent TBPVE+ TACE , among whom there were 11 cases with a history of HBV infection.Average age of the 13 patients was (60.8±6.2)years. The live function of all patients were Child-Pugh A classification.The CT or MRI images of each patient was reconstructed and the standard liver volume(SLV) before TBPVE+ TACE was (1 181.2±49.3)ml, estimated future liver remnant(FLR) was (326.1±72.1)ml and FLR/SLV was (27.6±6.0)%.The puncture site for TBPVE was determined by the three-dimensional reconstruction of portal vein.CT scan or MRI, AFP and liver function test were repeated after one and two weeks after TBPVE+ TACE.FLR and FLR/SLV were calculated respectively.Hepatectomy would be performed if the patients agreed.The postoperative complications were analyzed. Results: On the 7^thday after TBPVE+ TACE, the FLR/SLV was(42.6±8.0)% and the FLR increasement was(56.0±24.6)%.The level of AFP decreased from(87.9±81.8)μg/L to (29.7±20.9)μg/L.On the 14^thday after TBPVE+ TACE, the FLR/SLV was(45.8±6.2)% and the FLR increasement was(71.8±29.0)%.Four patients underwent surgery which including 2 right hepatectomies and 2 right trisegmentectomies in 2 weeks after TBPVE+ TACE.Nine patients were performed with targeting intratumoral lactic acidosis TACE (TILA-TACE). No severe complication occurred in all patients. Conclusions TBPVE could induce a rapid growth of the liver remnant but still with the concern of inducing the growth of neoplasms at the same time.To combine TACE in TBPVE therapy not also can the growth of neoplasms be prevented but also inducing its shrinking.This method might be a new mode for the treatment of hepatocellular carcinoma.

Gastric cancer (GC) is one of the most frequent malignant tumors. In order to systematically characterize the cellular and molecular mechanisms of intestinal GC development, in this study, we used 22 K oligonucleotide microarrays and bioinformatics analysis to evaluate the gene expression profiles of GC in 45 tissue samples, including 20 intestinal GC tissue samples,20 normal appearing tissues (NATs) adjacent to tumors and 5 noncancerous gastric mucosa tissue samples. These profiles allowed us to explore the transcriptional characteristics of GC and determine the change patterns in gene expression that may be of clinical significance. 1519 and 1255 differentially- expressed genes (DEGs) were identified in intestinal GC tissues and NATs, respectively, as determined by Bayesian analysis (P < 0.001). These genes were associated with diverse functions such as mucosa secretion, metabolism, proliferation, signaling and development, which occur at different stages of GC development.

Objective To observe the possible mechanism and inhibitory effects of curcumin on pulmonary fibrosis induced bleomycin in rats at the fibrosing stage. Methods 80 male Sprague-Dawley rats were random divided into 4 groups (20 rats in each group). Rats in the fibrosis model group, the prednisone group and the curcumin group were induced by instilled bleomycin through tracheal, rats in the control group with same volume normal saline. Since the 15th day after bleomycin administration, the curcumin group and prednisone group were given curcumin (300 mg/kg) or prednisone (5mg/kg) per day by intragastric administration, respectively. The normal control group and the model group were given 1% sodium carboxymethyl cellulose ( 10ml/kg). Six rats of each group were random sacrificed on the 21st, 28th, 42nd and 56th days after bleomycin administration. The histological changes of the pulmonary were evaluated by H. E and Masson dyeing. The expressions of transforming growth factor-β1 (TGF-β1), platelet-derived growth factor (PDGF) and hydroxyproline in the tissue of pulmonary were assessed by immunohistochemistry and digestion method. Results Pulmonary fibrosis and hydroxyproline level in the curcumin group were obviously reduced as compared with the model group on the 42nd and 56th day[42 d:1. 28 ±0. 61 vs 2. 28 ±0. 39,P <0. 01 ;(1.73 ±0. 22)mg/g vs (2.50 ±0. 37) mg/g, P <0.01;56 d:1.00 ±0.59 vs 1.73 ±0.36, P< 0. 05; ( 1.57 ± 0. 36) mg/g vs (2. 20 ± 0. 42) mg/g, P < 0. 01 ], and it was also lower than that in prednisone group on the 42nd day( P < 0. 05 ). The expression of TGF-β1 and PDGF in the curcumin group were obviously lower than that in the model group on the 28th, 42nd and 56th day[28 d:TGF-β1 :3642. 05 ±839. 31 vs 5067. 35 ±738. 39, P <0. 05 ;PDGF:2957. 55 ±739. 16 vs 4457. 75 ±568. 39, P <0. 05;42 d: TGF-β1: 2689. 73 ± 529.22 vs 4089. 50 ± 619. 37, P < 0. 01; PDGF: 2834. 46 ± 567. 16 vs 3239. 52 ±628. 26, P <0. 01 ;56 d:TGF-β1: 1968.57 ±408. 36 vs 2968.20 ±498.42, P <0. 01 ;PDGF: 1083.36 ±381.35 vs 2019. 40 ±412. 36, P <0. 01 ], which was lower than that in prednisone group on the 42nd and 56th day (42 d,TGF-β1 :3529. 07 ±981.35,PDGF:2618. 34 ±813. 34;56 d,TGF-β1 :2530. 83 ±439. 37,PDGF: 1738. 35 ±536. 62, Pall <0. 05 ) , and it had no obvious difference compared with control group on the 56th day ( P > 0. 05 ). Conclusion Curcumin could alleviate bleomycin-induced pulmonary fibrosis in rats at the fibrosing stage by inhibiting the expressions of TGF-β1 and PDGF.

Objective To observe the effects of CD4~+ CD25~+ regulatory T cells ( CD4~+ CD25~+ Treg) on the airway inflammation of asthmatic rats. Methods CD4~+ CD25~+ Treg of OVA- immune tolerance rats were transferred to asthmatic rats. Then bronchoalveolar lavage fluid (BALF) was collected, and cytology study was conducted. The IL-4, IL-5, IFN-γ and OVA-specific serum IgE level in BALF were determined by ELISA. The lung tissue was obtained, and histologieal analysis was done through H. E. Results Total cells number, the percentage of lymphocytes and neutrophils in BALF, the IL-4 and IL-5 BALF levels and the OVA-specific serum IgE level of adoptive transfer group were decreased ( P < 0.05 ) , and the percentage of eosinophils ( Eos) was significantly lower than that of asthma group ( P < 0.01) , while its BALF IFN-γ level was higher than that of asthma group( P <0. 05). Compared with that of asthma group, peribronchiole inflammatory of treated group was alleviated. Conclusion CD4 ~+ CD25~+ Treg of OVA- immune tolerance rats transferred to asthmatic rats can significantly alleviate the airway inflammation of asthmatic rats.