Objective:To construct pancreatic cancer cell lines expressing luciferase and mesothelin (MSLN), and evaluate the feasibility of using them as target cells in analyzing the cytotoxicity activity of immune cells.Methods:Lentiviral vectors expressing luciferase and MSLN genes were constructed, and pancreatic cancer cell lines were infected after lentivirus packaging. Single-cell clones were obtained by limited dilution following antibiotic screening, and the stable expression of the target genes were verified. These cells were used as target cells to detect the cytotoxicity of immune cells by real-time cell analysis (RTCA) and luciferase activity. Besides, these luciferase-expressing cells were transplanted into B-NDG mice to establish the animal models of pancreatic cancer, and in vivo optical imaging technology was used to detect the expression of luciferase and monitor the tumor growth in mice. The cytotoxicity of chimeric antigen receptor T (CAR-T) cells was verified in these animal models. Results:Three pancreatic cancer cell lines, panc-1-luc, panc-1-luc-MSLN and capan-2-luc, that could stably express luciferase and MSLN genes were successfully constructed. The expression of the reporter gene in these cells were high, and positively correlated with the number of cells. There were 95.6% of panc-1-luc-MSLN cells expressing MSLN. MSLN-CAR-T cells had specific killing effect on MSLN-positive panc-1-luc-MSLN cells and capan-2-luc cells, with the minimum killing rates of (70.00±18.19)% and (57.00±5.29)%, respectively. But they had no cytotoxicity to MSLN-negative panc-1-luc cells. RTCA results showed that MSLN-CAR-T cells were able to lyse all three pancreatic cancer cell lines, and the minimum killing rates were (56.33±7.64)%, (93.00±2.65)% and (26.33±28.15)%, respectively. The killing of target cells by NK-92MI cells was not depended on MSLN expression. The cytotoxicity in the mice models of pancreatic cancer was consistent with the results in vitro. The in vivo and in vitro test results suggested that the expression of luciferase by target cells could reflect the cytotoxicity of immune cells. Conclusions:This study establishes three pancreatic cancer cell lines stably expressing luciferase, which can be used to evaluate the cytotoxicity of immunotherapy products targeting tumor cells in vitro and in vivo.

Purpose To investigate the clinical,imaging,pathological,and genetic characteristics of neuroepithelial tumors with EWSR1 translocation.Methods The clinicopatho-logical data of 6 patients with EWSR1 translocation in neuroepi-thelial tumors were collected,routine HE and immunohistochem-ical staining were performed,the information of high-throughput sequencing was summarized,and the relevant literature was re-viewed.Results The median age of the 6 patients was 11.5 years(ranging from 1.9 to 17 years),including 1 male and 5 females.The tumors located in temporal lobe,frontal lobe,pari-etal lobe,suprasellar region,or lateral ventricle.The clinical manifestations mainly started with seizures.Brain MRI showed abnormal signal focus in the cerebral hemisphere near the cortex in 4 cases,and ventricle/periventricular regions in 2 cases,with an almost clear boundary in 5 cases.Microscopically,the histo-logical changes were diverse,including low-grade gliomas/gli-oneuronal tumors in 3 cases,high-grade gliomas in 2 cases,and glioneuronal tumor with high-grade feature in 1 case.Immuno-histochemically,tumor cells expressed GFAP,S-100,Syn,and Olig2 partially.2 cases exhibited slightly positive of NeuN and 1 case exhibited little dot-like staining of EMA.Next generation sequencing revealed EWSR1 rearrangement in all 6 cases,with chaperone genes including PATZ1 in 5 cases,and PLAGL1 in 1 case.3 cases were treated with chemotherapy after surgery,and no recurrence or progression was found during follow-up.Con-clusion The neuroepithelial tumors with the fusion of EWSR1 and non-ETS commonly occur in the cerebral hemisphere of teenagers and children.Most of the boundaries lesion are still clear,the histomorphological spectrum is diverse,and the bio-logical behavior is presented as a low to moderate malignancy,which provides the possibility for expanding the molecular classi-fication of CNS neuroepithelial tumor.

Objective To investigate the effect of mechanical thrombectomy using Solitaire stent combined with dual antiplatelet therapy(DAPT)on limb function and vascular reocclusion in patients with acute cerebral infarction.Methods A total of 134 patients with acute cerebral infarction who received treatment in our hospital from February 2020 to August 2023 were selected as samples.According to whether the patients regularly took aspirin and clopidogrel every day one week before surgery,they were divided into observation group(taking the medications)and control group(without taking medications),with 67 cases in each.The control group was given mechanical thrombectomy,and the observation group was treated with DAPT combined with mechanical thrombectomy.Outcomes measured included revascularization one week after treatment,and neurological and limb function assessment using NIHSS and FMA scale at baseline,1 day,1 week,and 1 month after treatment.Brain hemodynamics(CVR,Qmean,and Vmean)and thrombosis markers(TXB2,CD62P,and 6-keto-PGF1α)were evaluated pre-treatment and one month after treatment.Vascular reocclusion three months after treatment and its correlation with hemody-namics and thrombosis factors were analyzed.Results After treatment,higher revascularization rate,FMA score,Qmean,Vmean,and 6-keto-PGF1α level,and lower NIHSS score,CVR,TXB2,and CD62P level were found in the observation group(P<0.05).The incidence of vascular reocclusion was also lower in the observation group.Logistic analysis indicated that brain hemodynamic indicators and the level of thrombosis-related factors influenced vascular reocclusion occurrence(P<0.05).Conclusion Mechanical thrombectomy using Solitaire stent combined with DAPT helps the recovery of limb function in patients with acute cerebral infarction and reduces the risk of vascular reocclusion.

Objective:To evaluate the feasibility of 8E5 cells and CD19-CAR-Jurkat cells used as reference cells in the detection of lentiviral vector integration sites with different methods.Methods:Single clones of 8E5 cells and CD19-CAR-Jurkat cells were selected using limiting dilution method. Digital PCR was established to detect the copy number of HIV-1 in 8E5 cells and the copy number of CAR in CD19-CAR-Jurkat cells. High-throughput sequencing techniques (whole-genome resequencing, modified genome sequencing and probe hybridization capture) were used to detect integration sites in 8E5 cells and CD19-CAR-Jurkat cells, and optical genome mapping (OGM) technology was used for further confirmation.Results:Three clones of 8E5-D8 cells and six clones of CD19-CAR-Jurkat 2-6 cells were selected using the limiting dilution method. 8E5-D8 and CD19-CAR-Jurkat 2-6 were chosen as candidate cells based on their gene copy numbers detected by digital PCR and flow cytometry. These cells were then expanded and cryopreserved. Digital PCR showed that 8E5-D8 cells contained approximately 1 copy per cell, while CD19-CAR-Jurkat 2-6 cells contained approximately 13 copies per cell. High-throughput sequencing revealed one integration site in 8E5 cells and 13 integration sites in CD19-CAR-Jurkat cells, which matched the copy number detection results. All these integration sites were further confirmed at the submicroscopic level of chromosomes using OGM.Conclusions:Based on the insertion copy numbers and integration sites, 8E5-D8 cells and CD19-CAR-Jurkat 2-6 cells could be used as reference cells in further development of methods for detecting integration sites in CAR-T cell lentiviral vectors.

BACKGROUND: LY01005 (Goserelin acetate sustained-release microsphere injection) is a modified gonadotropin-releasing hormone (GnRH) agonist injected monthly. This phase III trial study aimed to evaluated the efficacy and safety of LY01005 in Chinese patients with prostate cancer. METHODS: We conducted a randomized controlled, open-label, non-inferiority trial across 49 sites in China. This study included 290 patients with prostate cancer who received either LY01005 or goserelin implants every 28 days for three injections. The primary efficacy endpoints were the percentage of patients with testosterone suppression ≤50 ng/dL at day 29 and the cumulative probability of testosterone ≤50 ng/dL from day 29 to 85. Non-inferiority was prespecified at a margin of -10%. Secondary endpoints included significant castration (≤20 ng/dL), testosterone surge within 72 h following repeated dosing, and changes in luteinizing hormone, follicle-stimulating hormone, and prostate specific antigen levels. RESULTS: On day 29, in the LY01005 and goserelin implant groups, testosterone concentrations fell below medical-castration levels in 99.3% (142/143) and 100% (140/140) of patients, respectively, with a difference of -0.7% (95% confidence interval [CI], -3.9% to 2.0%) between the two groups. The cumulative probabilities of maintaining castration from days 29 to 85 were 99.3% and 97.8%, respectively, with a between-group difference of 1.5% (95% CI, -1.3% to 4.4%). Both results met the criterion for non-inferiority. Secondary endpoints were similar between groups. Both treatments were well-tolerated. LY01005 was associated with fewer injection-site reactions than the goserelin implant (0% vs . 1.4% [2/145]). CONCLUSION: LY01005 is as effective as goserelin implants in reducing testosterone to castration levels, with a similar safety profile. TRIAL REGISTRATION ClinicalTrials.gov, NCT04563936.
Humans ; Male ; Antineoplastic Agents, Hormonal/therapeutic use* ; East Asian People ; Gonadotropin-Releasing Hormone/agonists* ; Goserelin/therapeutic use* ; Prostate-Specific Antigen ; Prostatic Neoplasms/drug therapy* ; Testosterone

Objective:Jue tone is a kind of sound,using 3-mi as the main tone,and is melodious,profound,makes people feel comfortable and pleasant.This study aimed to explore the probable mechanism of Jue tone to reduce blood pressure (BP) in hypertensive rats with a liver-fire hyperactivity pattern by observing changes in BP as well as physiochemical indexes in plasma.Methods:Sixteen male spontaneous hypertensive rats (SHR) with a liver-fire hyperactivity pattern were randomly divided into a control group and a Jue tone group.The rats in the Jue tone group were treated with Jue tone (55-65 dB,played by the five elements of music rhythm instrument,a kind of physio-therapy regimen music played by Shen Wu) once a day for 4 weeks.The BP levels in each group were measured twice a week,on Monday and Friday.The levels of angiotensin Ⅱ (Ang-Ⅱ),thromboxane B2(TXB2),endothelin-1 (ET-1),calcitonin gene-related peptide (cGRP),norepinephrine (NE),cortisol(CORT),and 5-hydroxytryptamine (5-HT).in plasma were detected by enzyme-linked immunosorbent assay after 4 weeks of intervention.Results:BP and the levels of TXB2 and ET-1 in the plasma of the treatment group were significantly lower than those of the control group,while the level of cGRP was significantly higher.Conclusion:Jue tone can reduce the BP of hypertensive rats with a liver-fire hyperactivity pattern.The mechanism may correlate with a reduction in TXB2 and ET-1 and an increase in cGRP with the reduction of reactive oxygen species (ROS).

Ankylosing spondylitis (AS) is a chronic inflammatory disease with early onset of reduced bone mineral density,which can lead to spinal deformity and even fracture,affecting patients′ quality of life seriously. To improve the compliance and long-term quality of life for AS patients, it is neccessary to enhance the awareness and knowledge of this disease among community primary health care providers.

ObjectiveTo investigate the effect of intraoperative Viatorr stent implantation for shunting of blood flow in the left or right branch of the portal vein on the clinical outcome of patients with cirrhotic portal hypertension undergoing transjugular intrahepatic portosystemic shunt (TIPS). MethodsA retrospective analysis was performed for the clinical data of 120 patients with cirrhotic portal hypertension who underwent TIPS in The General Hospital of Western Theater Command from March 2016 to December 2019, and according to the target position of portal vein puncture determined by intraoperative angiography, the patients were divided into left branch group and right branch group. The two groups were compared in terms of the incidence rates of postoperative recurrence and bleeding, stent dysfunction, and hepatic encephalopathy (HE) and survival. The t-test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. The Kaplan-Meier curve was used to calculate rebleeding rate, stent patency rate, incidence rate of HE, and survival rate. ResultsThe surgical success rate was 100% for all 120 patients, with a short-term hemostasis rate of 100%. Among the 120 patients, 52 underwent shunting of the left branch of the portal vein and 68 underwent shunting of the right branch. There was a significant reduction in portal venous pressure after surgery (9.98±2.84 mm Hg vs 24.72±5.11 mm Hg, t=37.76, P＜0.01). The cumulative rebleeding rates at 12 and 24 months after surgery were 3.2% and 11.0%, respectively, and the cumulative incidence rates of HE at 3, 6, 12, and 24 months after surgery were 10.8%, 13.6%, 21.2%, and 24.5%, respectively. Among the 29 patients who experienced HE, 23 had grade Ⅰ-Ⅱ HE and 6 had grade Ⅲ HE. The cumulative incidence rates of stent dysfunction at 12 and 24 months after surgery were 7.1% and 21.4%, respectively. The cumulative survival rates at 12 and 24 months after surgery were 92.0% and 86.5%, respectively. As for comparison of the left branch group and the right branch group, there were no significant differences in postoperative stent patency rate, rebleeding rate, incidence rate of HE, and survival rate(all P＜0.05). ConclusionTIPS is a safe and effective method for the treatment of cirrhotic portal hypertension, and intraoperative Viatorr stent implantation, no matter for establishing the shunt of the left or right branch of the portal vein, will not affect the clinical outcome of patients.

Transjugular intrahepatic portosystemic shunt (TIPS) is a minimally invasive technique for the treatment of portal hypertension in liver cirrhosis. With the gradual maturity of TIPS technique and the continuous improvement of stent materials, the indications for TIPS are gradually expanded, the incidence rate of complications gradually decreases, and the treatment outcome is getting better and better. The impact of TIPS on the main organs of patients has attracted more and more attention in recent years, and this article reviews the effect of TIPS on the liver and the heart.

Objective: To understand the awareness and influencing factors of medical healthcare APP among female college students in Weifang, and to provide references for healthy lifestyle and health literacy improvement. Methods: A total of 891 female college students were selected by stratified random sampling method, and were investigated regrading awareness on medical and health care APP and associated factors. Data were statistically analyzed by composition ratio, CHI-square test and binary Logistic regression. Results: There were 54.6% of the female college students who reported not aware of medical healthcare APP. The overall awareness rates of medical and health care APP for freshmen,sophomores, juniors, seniurs and above were 39.75%,45.59%,55.78% and 52.56%, respectively, and the difference was statistically significant(χ2=16.43，P<0.05). Logistic regression analysis showed that medical costs, medical healthcare information attention, medical health care information recognition, consulting the treatment scheme were positively associated with awareness of health care APP(OR=1.40,1.51,1.27,1.33,P<0.05). Conclusion The awareness rate of medical health care APP of female college students in Weifang is relatively low,and the difference between different groups is obvious. It is necessary to improve the scientific knowledge rate of the female college students to the APP, so as to influence the life style of the female college students.