AIM: To investigate and optimize the technology of ultrasonic-assisted extraction of arctiin and arctigenin from Fructus arctii.METHODS: The effects of ultrasonic power,particle size,ratio of liquid to solid,extraction time,extraction temperature and duty factor on the extraction rates of arctiin and arctigenin were investigated,and optimized with orthogonal experiments.RESULTS: The results showed that the optimal extraction conditions were as follows: particle 80 ～ 100 mesh 14 mL/g,ratio of liquid to solid 400 W ultrasonic power,50 ℃ of extraction temperature,and 10 min of extraction time.CONCLUSION: The optimized conditions are reasonable and reliable.

Objective To observe whether or not long-term inhalation of nitrous oxide can affect the cardiac function in rats. Methods Sprague-Dawley rats of either sex were randomly divided into four groups. Group A: inhaled common air for 50 days as control; Group B: inhaled 500mL/L N_2O for 15 days, two hours every day; Group C: inhaled 500mL/L N_2O for 30 days, two hours every day; Group D: inhaled 500mL/L N_2O for 50 days, two hours every day. Float glass intracellular microelectrode recording technique was used for observation of the duration (MAPD_(90), MAPD_(50) and MAPD_(20)) and amplitude of action potentials (APs) of left ventricular muscle cells in vitro. Angiotensin-2 (Ang-2) and eNOS were detected by immunohistochemical technique. Results ① Compared with that in Group A, the Aps duration of left ventricular muscle cells (MAPD) in Group B had no significant change while the MAPDs in Group C and Group D were extended significantly. There were no significant differences among the four groups in Aps amplitude. ② The expression of Ang-2 did not differ significantly between Group B and Group A. The expression level was higher in Group C and D than in Group A. ③ The expression level of eNOS was significantly lower in Group C and D than in Group A (P<0.05). Conclusion Long-term inhalation of N2O can significantly affect the cardiac function in rats, which may be related to higher expression of AngⅡin the heart induced by the long-time excitation of sympathetic nerves.

Examination paper analysis is an important job in examination management.It includes qualificative analysis and quantitative analysis,which are connective and complementary.This essay discusses the selection and application of examination paper analysis indexes.

Objective: To retrospectively analyze the efficacy and safety of modified cell infusion method in reducing the incidence of febrile non-hemolytic transfusion reaction (FNHTR). Methods: A total of 69 patients were enrolled in the clinical trial of CD₁₉ chimeric antigen receptor T (CAR-T) cell treatment from February 2017 to October 2018. Study group received the modified cell infusion method, that 1×10⁶ CAR-T cells were re-suspended in 2 mg human serum albumin with total volume of 20 ml and injected intravenously. The control group was intravenously administrated with CAR-T cell in 100 ml normal saline. The incidence of FNHTR, cytokine releasing syndrome (CRS) grade, cytokine level and efficacy were compared. Results: (1)The incidence of FNHTR in the study group was 21.1%, significantly lower than that in the control group (71%)(P=0.000). (2)There was no statistical difference in cell proliferation between the study group and the control group on day 4, 7, 14 and 21 after CAR-T cell infusion (P=10.223, 3.254, 5.551, 7.605). (3)There was no statistical difference in CRS grading between the study group and the control group (P=0.767). There was no statistical difference in the levels of interleukin 2 receptor (IL-2R), IL-6, tumor necrosis factor (TNF)-α between the two groups. (4)The C-reaction protein (CRP) level of the study group was lower than that of the control group on day 4 and 7 (P=0.026, 0.007). (5)There was no statistical difference of response rates in acute lymphocytic leukemia (ALL) and non-Hodgkin lymphoma (NHL) patients between the two groups (P_ALL=0.842; P_NHL=0.866). Conclusion The modified cell infusion method in CD₁₉ CAR-T cell treatment reduces the incidence of treatment-related FNHTR. It does not affect the proliferation of CAR-T cells in vivo, the grading of CRS and the response rates.

Objective: To observe the changes of PD-1 expression, mRNA level and cytotoxic activity of CD19 CAR-T cells during the culture process of CAR-T cells. Methods: The peripheral blood T cells of 6 lymphoma patients with high expression of PD-1 and 6 healthy volunteers were the source of CAR-T cells. The expression of PD-1 was analyzed by flow cytometry. The mRNA level of PD-1 was analyzed by PCR. The cell proliferation was analyzed by CCK-8 assay. The cytotoxicity was analyzed by LDH assay. Results: ①The transfection efficiency of high PD-1 expression T cells and healthy volunteer T cells were as the same (P>0.05) . ②The cell proliferation capacity of CD19 CAR-T cells from high PD-1 expression T cells or healthy volunteer T cells, with or without PD-1 inhibitor were as the same (P>0.05) . ③The cytotoxicity to lymphoma cells of high PD-1 expression T cells and CAR-T cells were lower than that of these two T cells combined with PD-1 inhibitor and the CAR-T cells from healthy volunteer T cells (P<0.001) . There was no difference of the cytotoxicity between the CAR-T cells from high PD-1 expression T cells combined with PD-1 inhibitor and the CAR-T cells from healthy volunteer (P>0.05) . ④There was no difference of the expression of PD-1 in all CAR-T cell groups during the culture process (P>0.05) . There was no difference of mRNA level of PD-1 in all groups during the culture process (P>0.05) . ⑤The PD-1 expression of CAR-T cells increased by the time of culture after contacting with lymphoma cells (P<0.001) . The PD-1 inhibitors could antagonize this effect. There was no difference of mRNA level of PD-1 in all groups after contacting with lymphoma cells (P>0.05) . Conclusion The PD-1 expression of CAR-T cells from high PD-1 expression T cells increased by the time of culture after contacting with lymphoma cells. However, the mRNA level of PD-1 of all groups did not change, even if PD-1 inhibitor was applied.

To retrospectively analyze the efficacy and safety of modified cell infusion method in reducing the incidence of febrile non?hemolytic transfusion reaction (FNHTR). Methods A total of 69 patients were enrolled in the clinical trial of CD19 chimeric antigen receptor T (CAR?T) cell treatment from February 2017 to October 2018. Study group received the modified cell infusion method, that 1×106 CAR?T cells were re?suspended in 2 mg human serum albumin with total volume of 20 ml and injected intravenously. The control group was intravenously administrated with CAR?T cell in 100 ml normal saline. The incidence of FNHTR, cytokine releasing syndrome (CRS) grade, cytokine level and efficacy were compared. Results (1)The incidence of FNHTR in the study group was 21.1%, significantly lower than that in the control group (71%)(P=0.000). (2)There was no statistical difference in cell proliferation between the study group and the control group on day 4, 7, 14 and 21 after CAR?T cell infusion (P=10.223, 3.254, 5.551, 7.605). (3)There was no statistical difference in CRS grading between the study group and the control group (P=0.767). There was no statistical difference in the levels of interleukin 2 receptor (IL?2R), IL?6, tumor necrosis factor (TNF)?α between the two groups. (4)The C?reaction protein (CRP) level of the study group was lower than that of the control group on day 4 and 7 (P=0.026, 0.007). (5)There was no statistical difference of response rates in acute lymphocytic leukemia (ALL) and non?Hodgkin lymphoma (NHL) patients between the two groups (PALL=0.842; PNHL=0.866). Conclusion The modified cell infusion method in CD19 CAR?T cell treatment reduces the incidence of treatment?related FNHTR. It does not affect the proliferation of CAR?T cells in vivo, the grading of CRS and the response rates.

Objective:To investigate the expression of c-Jun and MMP-9 in gastric cancer tissues,para-cancerous tissues and metastastic lymph nodes,and to explore its role and significance for the clinicopathology and prognosis.Methods:Immunohistochemistry was employed to detect the expression of c-Jun and MMP-9 in tissue microarrays containing gastric normal mucosa(n=32),para-cancerous tissues(n=54),metastastic lymph nodes(n=41),and gastric cancer tissues(n=189).Results:The positive rates for c-Jun and MMP-9 expression in gastric cancer were 73.0%and 78.3%,respectively.The positive rates of c-Jun protein was significantly associated with the degree of differentiation(P<0.05),but was not associated with the depth of invasion,lymph node metastasis,Lauren type,sex,age or size of tumor(P>0.05).The positive rates of MMP-9 was significantly associated with the depth of invasion,lymph node metastasis,Lauren type and degree of differentiation(P<0.05),but was not associated with sex,age or size of tumor(P>0.05).The positive rates of MMP-9 expression in the 41 gastnc cancer tissue samples and 41 metastastic lymph node tissue samples were significantly different(P<0.05).In metastastic lymph nodes,the positive rate of MMP-9 expression was higher.Kaplan-Meier survival analysis showed that the survival rate of patients with negative c-Jun and MMP-9 expression was higher than that of patients with positive c-Jun and MMP-9 expression(P<0.05).COX regression analysis showed that c-Jun and MMP-9 expressioh was not independent prognostic factor for gastric cancer. Conclusion:The expression of c-Jun is positively associated with the degree of differentiation.The increased c-Jun expression maybe an early indicator of gastric Cancer. The high expression of MMP-9 may involve the Occurrence,development,invasion,and metastasis of gastric cancer. C-Jun and MMP-9 are useful markers for predicting the outcome of gastric cancer,but they are not independent prognostic factors.

Objective:To compare the activity difference of the high affinity humanized CD19 chimeric antigen receptor (CAR)-T cells and murine CD19 CAR-T cells.Methods:Peripheral venous blood T cells from 8 healthy volunteers were collected and infected with humanized and murine CD19 CAR lentivirus. Human and murine CD19 CAR-T cells were prepared and cell proliferation was detected by cell counting kit-8 (CCK-8) method. The cytotoxicity of CD3 + T cells, humanized and murine CD19 CAR-T cells to NALM-6 cells was detected by lactate dehydrogenase assay. Thirty BAL B/c nude mice transplanted with NALM-6 cells were randomly divided into 3 groups with 10 mice in each group and injected humanized CD19 CAR-T cells, mouse CD19 CAR-T cells and control CD3 + T cell via tail vein, respectively. The proportion of NALM-6 cells in peripheral blood and the proportion of CD19 CAR-T cells in T cells from the vein of the inner canthus were detected by flow cytometry. The overall survival of BAL B/c nude mice was observed. Results:The proliferation of mouse and humanized CD19 CAR-T cells were (68.50±0.93)% and (80.63±1.41)%, respectively ( t=20.353, P<0.001) after cultured in vitro for 24 hours, and were (91.38±1.41)% and (148.13±1.25)%, respectively ( t=85.364, P<0.001) after cultured for 48 hours. When the effect to target ratio was 1∶1, there was no difference between the humanized and murine CD19 CAR-T cell group after co-culture for 24 hours ( P=0.169), while the killing activity of humanized CD19 CAR-T cells against NALM-6 cells was higher than that of murine CD19 CAR-T cells ( P<0.01) after 48 hours of co-culture. When the effect to target ratio was 4∶1, the cytotoxicity of humanized CD19 CAR-T cells against NALM-6 cells was higher than that of murine CD19 CAR-T cells in co-culture for 24 and 48 hours ( P<0.01). On the seventh day of CD19 CAR-T cell therapy, the proportion of NALM-6 cells in the peripheral blood of BAL B/c nude mice decreased to the lowest level in the humanized CD19 CAR-T cell group and the murine CD19 CAR-T cell group. After 21 days, the proportion of NALM-6 cells in the murine CD19 CAR-T cell group was higher than that in the humanized CD19 CAR-T cell group ( P21 d=0.001, P28 d<0.001, P35 d<0.001). The proportion of humanized and murine CD19 CAR-T cells in the peripheral blood reached the peaks after 7 days of therapy, and the proportion of humanized CD19 CAR-T cells was higher than that of murine CAR-T cells ( P7 d=0.002). The CD19 CAR-T cells disappeared in the peripheral blood in the murine CD19 CAR-T cell group after 14 days of therapy, while in the humanized CD19 CAR-T cell group it disappeared after 21 days of therapy. The median survival of BAL B/c nude mice in the murine CD19 CAR-T cell group and the humanized CD19 CAR-T cell group was 42 days and 63 days, respectively ( χ2=15.382, P<0.001). Conclusions:High affinity humanized CD19 CAR-T cells have stronger proliferation, higher cytotoxicity and longer survival time compared with those of murine CD19 CAR-T cells. The results indicate that the clinical efficacy of humanized CD19 CAR-T cells would be better than that of murine CD19 CAR-T cells.

Objective:To compare the activity difference of the high affinity humanized CD19 chimeric antigen receptor (CAR)-T cells and murine CD19 CAR-T cells.Methods:Peripheral venous blood T cells from 8 healthy volunteers were collected and infected with humanized and murine CD19 CAR lentivirus. Human and murine CD19 CAR-T cells were prepared and cell proliferation was detected by cell counting kit-8 (CCK-8) method. The cytotoxicity of CD3 + T cells, humanized and murine CD19 CAR-T cells to NALM-6 cells was detected by lactate dehydrogenase assay. Thirty BAL B/c nude mice transplanted with NALM-6 cells were randomly divided into 3 groups with 10 mice in each group and injected humanized CD19 CAR-T cells, mouse CD19 CAR-T cells and control CD3 + T cell via tail vein, respectively. The proportion of NALM-6 cells in peripheral blood and the proportion of CD19 CAR-T cells in T cells from the vein of the inner canthus were detected by flow cytometry. The overall survival of BAL B/c nude mice was observed. Results:The proliferation of mouse and humanized CD19 CAR-T cells were (68.50±0.93)% and (80.63±1.41)%, respectively ( t=20.353, P<0.001) after cultured in vitro for 24 hours, and were (91.38±1.41)% and (148.13±1.25)%, respectively ( t=85.364, P<0.001) after cultured for 48 hours. When the effect to target ratio was 1∶1, there was no difference between the humanized and murine CD19 CAR-T cell group after co-culture for 24 hours ( P=0.169), while the killing activity of humanized CD19 CAR-T cells against NALM-6 cells was higher than that of murine CD19 CAR-T cells ( P<0.01) after 48 hours of co-culture. When the effect to target ratio was 4∶1, the cytotoxicity of humanized CD19 CAR-T cells against NALM-6 cells was higher than that of murine CD19 CAR-T cells in co-culture for 24 and 48 hours ( P<0.01). On the seventh day of CD19 CAR-T cell therapy, the proportion of NALM-6 cells in the peripheral blood of BAL B/c nude mice decreased to the lowest level in the humanized CD19 CAR-T cell group and the murine CD19 CAR-T cell group. After 21 days, the proportion of NALM-6 cells in the murine CD19 CAR-T cell group was higher than that in the humanized CD19 CAR-T cell group ( P21 d=0.001, P28 d<0.001, P35 d<0.001). The proportion of humanized and murine CD19 CAR-T cells in the peripheral blood reached the peaks after 7 days of therapy, and the proportion of humanized CD19 CAR-T cells was higher than that of murine CAR-T cells ( P7 d=0.002). The CD19 CAR-T cells disappeared in the peripheral blood in the murine CD19 CAR-T cell group after 14 days of therapy, while in the humanized CD19 CAR-T cell group it disappeared after 21 days of therapy. The median survival of BAL B/c nude mice in the murine CD19 CAR-T cell group and the humanized CD19 CAR-T cell group was 42 days and 63 days, respectively ( χ2=15.382, P<0.001). Conclusions:High affinity humanized CD19 CAR-T cells have stronger proliferation, higher cytotoxicity and longer survival time compared with those of murine CD19 CAR-T cells. The results indicate that the clinical efficacy of humanized CD19 CAR-T cells would be better than that of murine CD19 CAR-T cells.

Objective:To explore the effects of bridge in, objective, pre-assessment, participatory learning, post-assessment and summary (BOPPPS) on internal nursing teaching.Methods:Totally 220 nursing students enrolled into Zhengzhou University in 2017 were selected by convenient sampling and divided into the observation group and the control group according to the teaching mode, with 110 nursing students in each group. Nursing students in the control group receiving traditional teaching, while nursing students in the control group receiving BOPPPS teaching. The effect of the two teaching modes was compared.Results:The satisfaction rate of the observation group (98.2%) was higher than that of the control group (90.0%) , and the difference was statistically significant ( P<0.05) . The assessment results of the observation group were higher than those of the control group, and the differences were statistically significant ( P<0.01) . The total score of self-evaluation of learning effect and the score of each dimension in the observation group were higher than those in the control group, and the differences were statistically significant ( P<0.05) . Conclusions:The BOPPPS model shows satisfying effects in internal nursing teaching, which can increase the satisfaction of students with the teaching mode, and enhance their interest in learning, academic performance, and learning evaluation.