Objective Toobservetheclinicalsignificanceofcerebrovascularhemodynamicindex (CVHI)inthepatientswithsilentcerebralinfarction.Methods Atotalof180patientsperformed cerebrovascular hemodynamics with ultrasound and head MRI examinations were enrolled retrospectively. They were divided into three groups:60 patients with silent cerebral infraction (SCI)were used as a SCI group,60 age-,sex- and previous medical history-matched high risk patients with cerebrovascular disease were used as a cerebrovascular disease risk factor (CV-HRF)group,and 60 healthy subjects were used as a normal control group over the same period. Cerebrovascular function detector was used to collect CVHI at the bilateral carotid arteries. The CVHI features of the 3 groups were analyzed and compared. Results Comparedwiththecontrolgroup,thereweresignificantchangesinvariousindexesofcerebral vascular hemodynamics in the SCI group and the CV-HRF group. The indexes of reflecting the cerebral blood supply state included significantly decreased mean blood flow,mean flow velocity,maximum flow velocity,and minimum flow velocity (P<0. 05);the indexes reflecting vascular elastic properties and resistance status included the increased pulse wave velocity,characteristic impedance,peripheral resistance,and dynamic resistance (P<0. 05);the indexes reflecting cerebral microcirculation included significantly increased critical pressure and decreased differential pressure (P<0. 05);the total score of cerebrovascular function was also decreased significantly,they were 44 ± 9,51 ± 5,and 85 ± 7,respectively (P<0. 05);there were significance differences in the distribution of total score amongthe3groups(P<0.05).Conclusion Thehigh-riskpatientswithsilentcerebralinfarction and cerebrovascular disease have the CVHI changes. The total integrated value of cerebrovascular function declined,and the former is more obvious.

Objective To establish the quality standards of Zizhu ointment. Methods The TLC was applied to identify Radix arnebiae and borneol of Zizhu ointment, and the content of borneol was determined by gas chromatography. Results The TLC spots were clear, well-separated and easy to identify. The good linear range of borneol reference substance on calibration curve was 0.048 4-1.210 0 μg, and the recovery was 90%-110%, the relative standard deviation was less than 5%. Conclusion The method is simple and feasible, can be used as the quality control method of Zizhu ointment.

Objective: To design and prepare a novel bi-specific chimeric antigen receptor (CAR)-T cell targeting both CD20 and CD19 antigen on B lymphocyte surface, and to detect its killing effect on B lymphocyte tumors as well as its treatment efficacy on immunodeficiency B-NSG mouse with leukemia. Methods: Bi-specific CAR molecule of CD20 (human originated)/CD19 (murine originated) scFv was constructed and packaged into lentiviral vector in 293 cells, and then transfected into T lymphocytes from healthy donors to prepare BiCAR-T cells. K562-CD19-GFP cells (with positive CD19 expression), K562-CD20-GFP cells (with positive CD20 expression) and Nalm6-Luc-GFP cells expressing luciferase were constructed as target cells. After being co-incubated with above mentioned targets cells, the cytotoxic effects of BiCAR-T cells on target cells were evaluated via LDH release assay, and the secretion of IFN-γ by BiCAR-T cells was evaluated by ELISA. Nalm6-Luc-GFP cells were used to construct the mouse model of leukemia and BiCAR-T cells were transfused via tail veins; the treatment efficacy of BiCAR-T cells on tumor bearing mice was evaluated with small animal imaging method. Results: After 7 days’incubation, the BiCAR-T cells originated from healthy donors amplified about 20-50 times with a positive rate of 10%~92%, indicating successful preparation of BiCAR-T cells. Under an effector∶target ratio of 10∶1, the killing rates of BiCAR-T cells against Nalm-6, K562-CD19-GFP and K562-CD20-GFP cells were significantly higher than that of control cells [(76.7±7.4)% vs (8.7±2.4)%, (93.3±5.2)% vs (46.7±6.2)%, (51.0±0.8) vs (30.7±0.5)%, all P<0.01]. Compared with control group, BiCAR-T cells co-incubated with Nalm-6 cells also secreted significantly more IFN- γ [(872.7±7.7) vs (101.0±5.3) pg/ml, P<0.01). Animal experiment showed that BiCAR-T cells had significant efficacy on B-NSG mice with leukemia; NSG leukemia mice treated with BiCAR-T cells all lived up to 70 days (till they were mercy killed) and leukemia cells disappeared at about 50 days, while the mice in PBS and T lymphocytes group all died at (19±3) d and (20±1) d, respectively. Conclusion: Bi-specific CAR molecules expressing CD19 and CD20 were successfully designed and BiCAR-T cells were successfully prepared. The BiCAR-T cells can effectively kill CD19 and/or CD20 tumor cells and secret large amounts of IFN-γ after co-incubation with target cells, exerting significant treatment efficacy on B-NSG immunodeficiency mouse with leukemia.