ABSTRACT
OBJECTIVE：To know about the co gnition level and self-medication behavior of antibiotics among urban and rural residents in Lu ’an city of Anhui province ，and to investigate its influential factors and to provide reference for promoting rational use of antibiotics. METHODS ：Totally 684 urban and rural residents aged 18-80 years in Lu ’an city were randomly selected as the research objects by stage sampling method. A self-designed questionnaire was used for household survey ，involving general demographic characteristics ，antibiotics related cognitive level ，antibiotics use behavior and related influential factors ，etc. RESULTS：A total of 657 questionnaires were collected ，with effective rate of 96.1％. Among them ，305 were from urban residents and 352 from rural residents. Among the 657 respondents，38.2％ were male and 61.8％ were female ；their age was （50.30±13.26）years old ；44.7％ of them were educated in primary school or below. 7.8％ of the respondents correctly recognized that antibiotics were not effective to the virus ；12.6％ knew antibiotic resistance ；55.1％ thought that frequent use of antibiotics would reduce the sensitivity of bacteria to it ；23.1％ said they knew the difference between prescription drugs and over-the-counter drugs；58.0％ could tell at least one case of not using antibiotics. For 7 knowledge items ，75.3％ of the residents in the survey area had a total score of less than 3；the cognition level of antibiotics was higher in urban areas and people with higher education level. 66.5％ of the respondents had used antibiotics in the past one year，of which 61.0％ obtained antibiotics by prescription 65161220。E-mail： from doctors ，50.7％ purchased antibiotics by themselves in pharmacies， and 13.1％ used the above two ways both. Among the people who have used antibiotics in the past year ， 81.9％ said they could buy antibiotics without prescription. Among the 657 respondents，49.0％ said that they had to obtain prescription from doctor when taking antibiotics ；68.9％ said that they would stop taking antibiotics when their symptoms improved；19.3％ would increase their dosage in order to enhance the curative effect ；28.3％ would change drugs frequently. Compared with urban residents ，rural residents were more likely to take antibiotics based on prescriptions by physicians [odds ratio （OR）＝1.693，95％ confidence interval （CI）（1.191，2.407）]. The higher the cognitive score ，the lower the behavior rate of having to prescribe antibiotics by doctors [OR ＝0.882，95％CI（0.785，0.991）]，and they were more likely to stop taking antibiotics when symptoms improved [OR ＝1.163，95％CI（1.025，1.319）]，and male were more inclined to increase the dosage of antibiotics to enhance the efficacy [OR ＝1.841，95％CI（1.214，2.792）]. The higher the cognitive score was ，the less likely they were to increase drug dosage to enhance the curative effect [OR ＝0.894，95％CI（0.773，1.034）]，nor were they inclined to change drugs frequently [OR ＝0.873，95％CI（0.767，0.992）]. CONCLUSIONS ：The cognition level to antibiotics of urban and rural residents in Lu ’an city needs to be improved urgently ，and reasonable antibiotic use behavior needs to be standardized. Pure knowledge of antibiotics is not necessarily related to the expected rational drug use behavior. Therefore ，in addition to health promotion for the rational use of drugs for residents ，it is also necessary to create a systematic environment that promotes the rational use of antibiotics and provide residents with multi-channel services for rational use of drugs.
ABSTRACT
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.
ABSTRACT
Objective@#To investigate the different functions of humanized and murinized CD19 chimeric antigen receptor (CAR)-T cells against Raji cell line in vitro and in vivo.@*Methods@#Peripheral blood samples were collected from eight patients with lymphoma who were going to receive CD19 CAR-T cell therapy and used for the preparation of peripheral blood mononuclear cells (PBMC) as well as humanized and murinized CAR-T cells. Cell proliferation and cytotoxicity were detected with CCK-8 and LDH assays, respectively. A tumor-bearing mouse model was established by injecting BALB/c female nude mice with fluorescent Raji cells. Changes in tumor volume in these mice were observed by in vivo imaging technology. The transfection efficiency and amount of CAR-T cells in the mice were detected with flow cytometry.@*Results@#No statistical difference in transfection efficiency was found between humanized and murinized CAR-T cells, nor in cell proliferation at 24 h of culture in vitro(P=0.104). The proliferation of humanized CAR-T cells showed a significant increase compared with that of murinized CAR-T cells at 48 h of culture (P=0.009). Similarly, the cytotoxicity of the two types of CAR-T cells against Raji cells showed no significant difference at 24 h at any effector/target (E/T) ratio (1∶1 or 4∶1), and that of humanized CAR-T cells was higher than that of murinized CAR-T cells at both E/T ratios at 48 h (E/T ratio=1∶1, P=0.005; E/T ratio=4∶1, P=0.008). Moreover, the cytotoxicity of CAR-T cells was higher than that of PBMC in any case. Tumor volumes in mice were reduced 14 d after humanized or murinized CAR-T cell therapy, while the mice in the PBMC control group suffered tumor progression. Tumor volume began to increase in mice 21 d after murinized CAR-T cell therapy, while no significant change was observed in the mice treated with humanized CAR-T cells. All of the mice died 25 d after murinized CAR-T cell therapy, while the deaths among those under humanized CAR-T cell therapy occurred on 31 d. The proportion of CAR-T cells in mice reached the peak 7 d after receiving humanized or murinized CAR-T cell therapy, while that in the humanized group was significantly higher than that in the murinized group at any time point (P4 d=0.001, P7 d=0.000, P14 d=0.003). Murinized CAR-T cells became undetectable on 21 d, while humanized CAR-T cells on 35 d. The maximum survival time for mice in the PBMC and murinized and humanized CAR-T cell groups was 20 d, 25 d and 53 d, respectively.@*Conclusions@#Compared with murinized CD19 CAR-T cells, humanized CD19 CAR-T cells showed stronger proliferation potential and cytotoxicity and remained in vivo detectable for a longer period of time. This study indicated that humanized CD19 CAR-T cells were superior to murinized CD19 CAR-T cells for the treatment of B cell lymphoma.
ABSTRACT
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 (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.
ABSTRACT
Objective To investigate the different functions of humanized and murinized CD19 chimeric antigen receptor ( CAR)-T cells against Raji cell line in vitro and in vivo. Methods Peripheral blood samples were collected from eight patients with lymphoma who were going to receive CD19 CAR-T cell therapy and used for the preparation of peripheral blood mononuclear cells ( PBMC) as well as humanized and murinized CAR-T cells. Cell proliferation and cytotoxicity were detected with CCK-8 and LDH assays, respectively. A tumor-bearing mouse model was established by injecting BALB/c female nude mice with flu-orescent Raji cells. Changes in tumor volume in these mice were observed by in vivo imaging technology. The transfection efficiency and amount of CAR-T cells in the mice were detected with flow cytometry. Re-sults No statistical difference in transfection efficiency was found between humanized and murinized CAR-T cells, nor in cell proliferation at 24 h of culture in vitro(P=0. 104). The proliferation of humanized CAR-T cells showed a significant increase compared with that of murinized CAR-T cells at 48 h of culture ( P=0. 009). Similarly, the cytotoxicity of the two types of CAR-T cells against Raji cells showed no significant difference at 24 h at any effector/target (E/T) ratio (1 : 1 or 4 : 1), and that of humanized CAR-T cells was higher than that of murinized CAR-T cells at both E/T ratios at 48 h (E/T ratio=1 : 1, P=0. 005;E/T ratio=4 : 1, P=0. 008). Moreover, the cytotoxicity of CAR-T cells was higher than that of PBMC in any case. Tumor volumes in mice were reduced 14 d after humanized or murinized CAR-T cell therapy, while the mice in the PBMC control group suffered tumor progression. Tumor volume began to increase in mice 21 d after murinized CAR-T cell therapy, while no significant change was observed in the mice treated with hu-manized CAR-T cells. All of the mice died 25 d after murinized CAR-T cell therapy, while the deaths among those under humanized CAR-T cell therapy occurred on 31 d. The proportion of CAR-T cells in mice reached the peak 7 d after receiving humanized or murinized CAR-T cell therapy, while that in the humanized group was significantly higher than that in the murinized group at any time point (P4 d=0. 001, P7 d=0. 000, P14 d=0. 003). Murinized CAR-T cells became undetectable on 21 d, while humanized CAR-T cells on 35 d. The maximum survival time for mice in the PBMC and murinized and humanized CAR-T cell groups was 20 d, 25 d and 53 d, respectively. Conclusions Compared with murinized CD19 CAR-T cells, humanized CD19 CAR-T cells showed stronger proliferation potential and cytotoxicity and remained in vivo detectable for a longer period of time. This study indicated that humanized CD19 CAR-T cells were superior to murinized CD19 CAR-T cells for the treatment of B cell lymphoma.
ABSTRACT
Objective To investigate the immunophenotypic characteristics of potential leukemia cells transfected with CD19 antigen receptor( CAR) during CAR-T cell preparation. Methods Morphological chan-ges in CD19 CAR-transfected cells were observed under inverted microscope. The transfection rate and immuno-phenotype of transfected Nalm-6 cells were analyzed by flow cytometry. Secretion of cytokines in the culture sys-tem was detected by chemiluminescence. Results The transfection rate of Nalm-6 cells by CD19 CAR was (46. 50±3. 78) ％ and that of KG1a cells was (15. 70±1. 22) ％. CD19 CAR-transfected Nalm-6 cells prolifer-ated more rapidly than Nalm-6 cells ( P values on 0 d, 4 d, 7 d and 12 d were 6. 339, 3. 447, 0. 012 and 0. 009). In the culture of CD19 CAR-transfected Nalm-6 cells, cell aggregation and adhesion were observed and they gradually gathered into a group. The rate of CD19 expression was only 1. 19％ in the CD19 CAR-transfect-ed Nalm-6 cell culture system with the transfection rate of (46. 50±3. 78) ％. After increasing the proportion of Nalm-6 cells in the culture system, CD19 expression was gradually increased, while the expression of CD22 re-mained stable. CD19 expressed by Nalm-6 cells cultured in the supernatant of CD19 CAR-transfected Nalm-6 cell culture system was decreased gradually. The levels of IL-10 and TNF-αsecreted by CD19 CAR-transfected Nalm-6 cells were higher than those by Nalm-6 cells. Conclusions Results of the immunophenotypic analysis of CD19 CAR-transfected leukemia cells suggested that CD22 CAR-T cell therapy could be used as a rescue or combination therapy for CD19 CAR transfection into leukemia cells.
ABSTRACT
Objective@#To investigate the efficacy and safety of CD19 chimeric antigen receptor T (CAR-T) lymphocytes for the treatment of B cell lymphoma.@*Methods@#A total of 22 patients with B-cell lymphoma from February 1, 2017 to July 1, 2018 were reviewed to evaluate the efficacy and adverse reactions of CD19 CAR-T.@*Results@#Of 22 patients with B-cell lymphoma received CD19 CAR-T cells, the median dose of CAR-T cells was 7.2 (2.0-12.0) ×106/kg. Nine of 12 cases of relapse refractory patients were overall response. Complete remission (CR) occurred in 2 of 12 patients, partial remission (PR) in 7 of 12 patients. The overall response in minor residual disease positive (MRD) group was 8 of 10 patients. CD19 CAR-T cells proliferated in vivo and were detectable in the blood of patients. The peak timepoints of CAR-T cells proliferated in the relapsed refractory and MRD positive groups were 12 (5-19) and 4.5 (1-12) days after treatment respectively, and among peripheral blood cells, CAR-T cells accounted for 10.10% (3.55%-24.74%) and 4.02% (2.23%-28.60%) of T lymphocytes respectively. The MRD positive patients achieved sustained remissions during a median follow-up of 8 months (rang 3-18 months) . None of all the patients relapsed during a median follow-up time of 10 months (3-18 months) . However, 7 PR responders of the relapsed refractory patients maintained a good condition for 1.5-6.0 months. One patient bridged to hematopoietic stem cell transplantation, another one sustained remission for 12 months. Cytokine-release syndrome (CRS) occurred in 14 patients with grade 1-2 CRS in MRD positive group and grade 3 CRS in relapsed refractory group.@*Conclusions@#CAR-T cell therapy not only played a role in the rescue treatment of relapsed and refractory patients, but also produced a surprising effect in the consolidation and maintenance of B-cell lymphoma. CD19 CAR-T cells might be more effective in the treatment of MRD positive B-cell lymphoma patients than in the refractory or relapsed cases. High response rate was observed with fewer adverse reactions.
ABSTRACT
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.
ABSTRACT
Objective@#To Evaluation the effect of PD-1 inhibitor Nivolumab on the proliferation and cytotoxicity of anti-CD19 chimeric antigen receptor T cells (CD19-CAR-T) in vitro.@*Methods@#Five patients with high PD-1 expression in peripheral blood and five healthy volunteers were selected. These peripheral blood mononuclear cells were used as the source of T cells to prepare CD19-CAR-T cells. Different doses (72, 36, 18 μg/ml) of Nivolumab was added on day 8 to the culture medium. Patient T cells incubated with 72 μg/ml Nivolumab and CD19-CAR-T cells of healthy volunteers were used as controls. CCK-8, lactate dehydrogenase (LDH) cytotoxicity assay and ELASA were used to detect the proliferation capacity, the specific cytotoxicity and the inflammatory factor secretion.@*Results@#①T cells from patients with high expression of PD-1 as the source of CD19-CAR-T cells did not affect transfection rate compared with that of healthy volunteers [(32.80±7.22)% vs (35.10±5.84)%, t=-0.554, P=0.593]. ②Incubation of CD19-CAR-T cells with 72 μg/ml Nivolumab did not affect CD19-CAR-T cell proliferation, but its cytotoxicity was significantly higher than that of CD19-CAR-T cells alone or patients’ T cells +72 μg/ml Nivolumab (all P<0.001), there was no significant difference in the killing activity between the 72 μg/ml and 36 μg/ml Nivolumab treated CD19-CAR-T cells on Pfeiffer cells (P=0.281, 0.267, respectively), and they were all higher than those of 18 μg/ml Nivolumab treated CD19-CAR-T cells (all P<0.001). ③Different doses of PD-1 inhibitor Nivolumab combined with CD19-CAR-T cells does not affect the secretion of IFN-γ and IFN-α (all P>0.05).@*Conclusion@#Combination of 36 μg/ml PD-1 inhibitor and CD19-CAR-T cells could reduce the drug toxicity and enhance the cytotoxicity.
ABSTRACT
Objective To investigate the effects of Bucinnazine Hydrochloride on the pain behavior and the expression of caveolin-1 (Cav-1) in the anterior cingulate cortex of neuropathic pain mice.Methods 64 adult male Kunming mice (20-25g) were divided randomly into 4 groups with 16 in each group:Sham+BH(Bucinnazine Hydrochloride) group,Sham+NS (Normal Saline) group,CCI+ BH group and CCI+ NS group.The corresponding drugs were administered by intraperitoneal injectionfrom the forth day after CCI once a day for three days.Paw thermal withdrawal latency was measured by Hargreaves methods.Mechanicalwithdrawal threshold was assayed by electronic dolorimeter.c-Fos protein in anterior cingulate cortex was detected by immunohistochemistry staining and the expression of t-Cav-1,p-Cav-1was detected by Western blot.Results Bucinnazine Hydrochloride administered by intraperitoneal injection(0.1 mg/10 g,mice) alleviated thermal hyperalgesia and mechanical allodynia of CCI mice.Compared with the forth day (4.92±0.41) s of CCI+BH group,paw withdrawal latency on the fifth day(5.92±0.61) s was increased(P＜0.05),and on the sixth day(7.93±0.91) s and seventh day (9.12±0.69)s were increased more(P＜0.01,P＜0.01).The paw withdrawal mechanical threshold on the sixth and seventh day of CCI+BH group mice((2.54 ±0.41)g,(3.68±0.61)g) were increased significantly (P＜0.01,P＜0.01)compared with the forth day(1.55± 0.31)g.Immunohistochenistry results showed that the expression of c-Fos decreased after treated with Bucinnazine Hydrochloride in the anterior cingulate cortex of CCI mice(P＜0.001).Western Blotting showed that the expression of t-Cav-1 (1.97±0.31) and p-Cav-1 (0.11 ±0.09) in the anterior cingulate cortex of CCI +BH group mice decreased compared with that of in CCI+NS group mice(t-Cav-1:2.87±0.15,p-Cav-1:0.48± 0.09) (P＜0.01,P＜0.01).Conclusion Bucinnazine Hydrochloride can alleviate both thermal hyperalgesia and mechanical allodynia of neuropathic pain of mice,and reduce the expression of c-Fos,t-Cav-1,p-Cav-1 in the anterior cingulate cortex of neuropathic pain mice.
ABSTRACT
Objective To study the correlation between expression level of als3 gene and the in vivo biofilm formation of Candida albicans in mice.Methods The real-time polymerase chain reaction (PCR) assay was used to detect als3 gene expressions of the clinical Candida albicans isolates from February 2016 to August 2016 in Tianjing No.1 Central Hospital.According to the expression levels of als3 gene, Candida albicans isolates were divided into high and low-expression groups.Thirty C57 mice were randomly assigned to high-expression group (n=15), low-expression group (n=5) and blank group (n=5).Animal model of Candida albicans biofilm was established based on venous catheter and intraperitoneal injection of Candida albicans.Catheters were removed after two weeks;inverted microscope was used for the observation of Candida albicans biofilm formation and transmission electron microscope was used for the observation of its ultrastructure.After irrigating the catheter, the growth of Candida albicans was observed;real-time PCR was used to detect the expression levels of als3 gene 12, 24, and 48 h after the catheter being removed.In this study, t test was used for measurement data and chi-square test was used for rate comparisons.Results In high-expression group, 11 strains (11/15) formed biofilms.In als3 low-expression group, only one strain (1/10) formed biofilm.The difference between these two group was statistically significant (x2=9.64,P0.05).In the als3 high-expression group, the expression of als3 gene declined gradually during the biofilm formation.In the als3 low-expression group, the change of als3 gene expression was not obvious.The expressions of als3 gene over time between two groups were significantly different (t=8.7, 10.3 and 9.2, respectively, all P<0.05).Conclusion The high expression of als3 gene in Candida albicans facilitates the formation of biofilm in vivo.
ABSTRACT
Objective To compare the expression of serum miR-100 in patients with esophageal cancer and healthy person ,and explore the value of miR-100 in diagnosis for esophageal cancer .Methods Real-time fluorescent quantitative polymerase chain reac-tion was used to detecting miR-100 in 40 esophageal cancer patients(study group) and 50 healthy person(control group) .Results The expression of miR-100 in the study group and control group were 6 .399 ± 3 .541 ,2 .625 ± 1 .515 respective ,the expression in the study group was significant higher than that of the control group(t= 9 .07 ,P< 0 .05) .The under area of receiver operating char-acteristic curve of miR-100 in diagnosis for esophageal cancer was 0 .832(95% confidence interval was 0 .731 - 0 .934) ,when the Cut off value was 5 .285 ,the sensitivity and specificity of miR-100 in diagnosis for esophageal cancer were 65% and 95% . Conclusion Serum miR-100 in esophageal cancer patients is higher than that in healthy person ,which might be a new molecular markers in diagnosis for esophageal caner .