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Objective:To explore the expression of miR-6883-5p in the serum of bladder cancer patients and its effect on the migration and proliferation ability of bladder cancer cells.Methods:The real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) method was used to detect the expression level of miR-6883-5p in the serum of 39 patients with bladder cancer and 39 healthy subjects, as well as the expression level of miR-6883-5p in normal bladder mucosal epithelial cells and bladder cancer cells. The bladder cancer cells with the lowest expression of miR-6883-5p were transfected with miR-6883-5p mimics or negative control (NC), namely miR-6883-5p group and NC group. Transwell migration experiment and cell counting kit-8 (CCK-8) colorimetric method were used to detect the migration and proliferation ability of cells transfected with miR-6883-5p. Bioinformatics software and dual luciferase reporter gene were used to predict and test the target genes of miR-6883-5p. qRT-PCR and Western blot were used to detect the expression levels of target genes in cells transfected with miR-6883-5p.Results:The expression level of miR-6883-5p in the serum of bladder cancer patients was significantly lower than that of healthy persons ( P<0.01). The expression level of miR-6883-5p in bladder cancer cells was significantly lower than that in normal bladder mucosal epithelial cells (all P<0.05), and the lowest expression of miR-6883-5p was in 5637 cells ( P<0.01). After transfection with miR-6883-5p, the migration and proliferation of cells in miR-6883-5p group were significantly lower than those in NC group (all P<0.05). Phosphatidyl alcohol proteoglycan-6 (GPC6) was the target gene of miR-6883-5p. After transfection with miR-6883-5p, the expression levels of GPC6 mRNA and protein in miR-6883-5p group significantly decreased ( P<0.01). Conclusions:miR-6883-5p is significantly low expressed in the serum of bladder cancer patients, and miR-6883-5p is related to the occurrence and development of bladder cancer; overexpression of miR-6883-5p can inhibit the migration and proliferation of bladder cancer cells; miR-6883-5p can inhibit the occurrence and development of bladder cancer by down-regulating the expression of GPC6.
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OBJECTIVE@#To observe the effect of acupuncture-moxibustion on negative emotions and plasma tryptophan (Trip)-kynurenine (Kyn) metabolism in the patients with Crohn's disease (CD) at the mild and moderate active stage.@*METHODS@#A total of 66 CD patients were randomized into an observation group (33 cases, 1 case dropped off) and a control group (33 cases, 2 cases dropped off). In the observation group, acupuncture was applied in combination with moxibustion. In the control group, the sham-acupuncture was used in combination with sham-moxibustion. In both of the observation group and the control group, acupuncture was applied to Zhongwan (CV 12), Shangjuxu (ST 37), Sanyinjiao (SP 6), Gongsun (SP 4), Hegu (LI 4), Quchi (LI 11), Taixi (KI 3) and Taichong (LR 3), and moxibustion was applied to Tianshu (ST 25) and Zusanli (ST 36). The treatment was given once every two days, 3 times a week, totally for 12 weeks. Separately, before and after treatment, the score of the hospital anxiety-depression scale (HADS) and the score of intestinal core symptoms (degree of abdominal pain and frequency of diarrhea) were observed in the patients of the two groups. The concentration of plasma indoleamine 2,3-dioxygenase 1 (IDO1) and the ratios of Kyn/Trp, QuinA/Kyn, KynA/Kyn and KynA/QuinA were compared between the two groups.@*RESULTS@#Compared with before treatment, the scores of HADS-A and HADS-D in the observation group and the score of HADS-A in the control group were all reduced after treatment (@*CONCLUSION@#Acupuncture and moxibustion relieve the negative emotions of anxiety and depression in CD patients at mild and moderate active stage, which is probably related to the regulation of plasma Trp-Kyn metabolic pathway.
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Humans , Acupuncture Points , Acupuncture Therapy , Crohn Disease/therapy , Emotions , Moxibustion , Plasma , Treatment Outcome , TryptophanABSTRACT
Objective To forecast the trend of mosquito density index in Pudong New Area, Shanghai so as to provide evidence for disease control and risk-control measures for vector-borne diseases. Methods Mosquito monitoring data was collected in Pudong New Area between 2011 and 2015 at the city-level monitoring sites for analysis on the trend of the mosquito density index in Pudong New Area of Shanghai by using the Autoregressive Integrated Moving Average Model (ARIMA). Results From 2011 to 2015, a total of 135 times labor-hour monitoring were carried out at the city-level monitoring points in Pudong New Area.The mosquito density index averaged 6.17/labor-hour with a standard deviation at 4.93, S=[0, 18]/labor-hour.Using ARIMA to analyze the change trend of mosquito density index in Pudong New Area, ARIMA(2, 0, 1)became the final fitting model, with R2=0.808.In the model, the Ljung-Box Q test value was 19.632(AR1=1.866, AR2=-0.907), and MA parameter was 0.999. Conclusion ARIMA model can be used to predict mosquito density monitoring data, but low monitoring frequency and irregular cycle length will affect the prediction results.
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Objective To learn the density, distribution, population composition, and seasonal fluctuation of mosquitoes in Shanghai Pudong New Area, and to provide evidence for the mosquito-borne diseases control and prevention. Methods From 2017 to 2019, light trap method was used to monitor the mosquito density in farmyard, parks, hospitals, residential areas and the barns, three times a month from April to November. Results A total of 24 times/set light traps were set up and 7 104 mosquitoes were captured.The average density was 3.07 insects per lamp.Culex pipienspallens (81.52%) and Aedes albopictus (12.10%) were the mostly captured, followed by Armigeres subalbatus (3.24%), Culex tritaeniorhynchus (2.78%) and Anopheles sinensis (0.36%).The proportion of Culex pipienspallens was increasing year by year, and that of Aedes albopictus was decreasing year by year.The seasonal variation curve of mosquito density showed a single peak distribution, with the peak from June to July.Among the five habitat types, the mosquito density from high to low was hospital (4.68/set), farmyard (4.38/set), livestock shed (3.25/set), residential household (2.81/set) and park (1.38/set).The mosquito density in urban area, urban-rural junction and rural area was 2.69, 3.32 and 3.36 respectively (i.e., rural area > urban-rural integration > urban area). Conclusion Culex pipienspallens and Aedes albopictus are the dominant species in Pudong New Area.Parks, hospitals and residential areas are the most important mosquitoes infestation areas, therefore more attention must be paid to mosquito control in these areas.
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Objective@#To construct the third generation chimeric antigen receptor based on a novel humanized anti-HER2 H1-2 scFv, and to investigate the specific cytotoxicity of H1-2 CAR modified T lymphocytes(CAR-T) against HER2+ tumor cells.@*Method@#The expression cassette of the third generation CAR gene and anti-HER2 H1-2 scFv were constructed and cloned into lentivirus transfer plasmid, and then the third generation H1-2 CAR was transduced into human T lymphocytes using lentivirus.Enzyme linked immunosorbent assay was used to detect the expression of cytokines IL2, and LDH release assay was used to detect the cytotoxic effect of the H1-2 CAR-T.Finally, NOD/SCID mice and HER2+ breast cancer cell line SKBR3 were used to detect the anti-tumor effect of H1-2 CAR-T in vivo.@*Results@#The third generation H1-2 CAR was successfully constructed.H1-2 CAR-T secreted high dose of IL2 after confrontation with HER2+ breast cancer cells.In vitro, the cytolytic rate of H1-2 CAR-T on high expression HER2+ tumor cells was significantly higher than that in low expression HER2 or non-expression HER2 tumor cells. At the efficacy to target ratio of 20, the cytolytic rate of H1-2 CAR-T against breast cancer cell SK-BR-3 could reach (90.1±2.8)%, while the cytolytic rate of H1-2 CAR-T against HER2- breast cancer cell MDA-MB-231 was only (13.5±4.7)%. In the mouse xenograft tumor model, H1-2 CAR-T cells inhibited breast cancer growth in vivo.At the end of the experiments, the average tumor weight in the H1-2 CAR-T cell treatment group was (0.7±0.1) g, the non-transfected T cell therapeutic group was (1.2±0.2) g, and the PBS group was (1.2±0.2) g. There was significant difference between the H1-2 CAR-T therapeutic group and the non-transfected T cell therapeutic group (P<0.05). However, there was no significant difference between the non-transfected T cell therapeutic group and the PBS treatment group (P>0.05).@*Conclusion@#The HER2-sepcific H1-2 CAR-T cells specifically kill HER2 positive cells, and further studies on CAR-T cells for the treatment of HER2+ cancers are useful.
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Objective:To artificially design and express a recombinant protein named as ScFv-pLLO by fusing ScFv gene of Rituximab(C2B8)and dominant antigen epitopes from listeriolysin O(LLO),and studying its anti-tumor activity.Methods:VH and VL gene sequences of C2B8 against CD20 were acquired by searching United States Patent database,and ScFv sequence was constructed by linking VL and VH with a short peptide linker.Two CD4+T cell epitopes from LLO were selected and designed to splice ScFv sequence.The recombinant gene of ScFv-pLLO was cloned into prokaryotic expression vector and purified after induction.The capacity of ScFv-pLLO target-binding to B-cell lymphomas was evaluated by flow cytometry ( FCM ) and co-immunoprecipitation ( Co-IP ) .The effects of ScFv-pLLO on B-cell lymphomas proliferation and apoptosis were detected respectively.The immunogenicity of ScFv-pLLO was assessed by lymphocyte proliferation assay.Results: ScFv-pLLO was successfully expressed.It could bind to different B-cell lymphomas cell lines and obviously inhibit the growth of Raji cells as well as inducing apoptosis.Moreover,ScFv-pLLO was able to stimulate proliferation of spleen lymphocytes of immunized mice.Conclusion: The recombinant protein ScFv-pLLO can target-bind to B-cell lymphomas,and perform inhibitory effect and induce apoptosis on Raji cells that indicate ScFv-pLLO retain the capacity of ScFv derived from monoclonal antibody against CD20.Besides, ScFv-pLLO can induce immune response.This study provides a basis for further research about the role of ScFv-pLLO on simulating tumor cell antigens as well as being tumor vaccine adjuvant.
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Objective To explore the structure of self-developed "The Compliance Scale Among Kidney Transplantation Recipients" in order to provide effective tool to evaluate kidney transplantation recipients' compliance. Methods 886 follow-up kidney transplantation recipients of six organ transplantation centers in Shanghai were surveyed by "The Compliance Scale Among Kidney Transplantation Recipients". Results The exploratory factor analysis yielded a 25-item four-factor model termed medication compliance, life habits compliance, self-monitoring compliance, follow-up compliance. Conclusions The self-developed "The Compliance Scale Among Kidney Transplantation Recipients" could be a useful tool for evaluating compliance among kidney transplantation recipients.
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Objective To know the advantages and disadvantages of problem-based learning from students'angle. Methods Interviewed 15 students by PBL scale and face to face interview,to know their ideas about problem-based learning,and then analyzed the results. Results PBL contributed to developing problem-solving skills,self-directed learning,team collaboration and extending professional knowl-edge. But it failed to show positive effect on students'learning motivation. Conclusions PBL has two sides on the outcomes of learning. Implementing PBL appropriately could gain the optimal effects.
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<p><b>BACKGROUND</b>Estrogen deficiency causes atrophic changes within the urogenital tract, and is associated with urinary symptoms. The purpose of this study was to investigate the effects of estrogen and tibolone on bladder histology, and the changes of estrogen receptor alpha and beta (ERalpha and beta) protein expression in the detrusor muscle.</p><p><b>METHODS</b>Forty female rats were separated into four groups of ten each. They received a sham operation (Sham), ovariectomy (Ovx), ovariectomy plus estrogen replacement (Ovx + E), or ovariectomy plus tibolone treatment (Ovx + T). After 12 weeks each rat was anesthetized and the bladders were removed. The bladders' ultra structure, collagen fiber (CF) to smooth muscle (SM) ratio and ER subtypes were studied. Statistical analyses were performed using the one-way analysis of variance test.</p><p><b>RESULTS</b>Ovx resulted in significant degeneration in bladder ultra structure; however, estrogen and tibolone reversed those changes. Ovx increased the CF/SM ratio, estrogen and tibolone resulted in an increase. Two estrogen receptors (ERs) were expressed in the bladder detrusor, with ERbeta the main subtype. Ovx resulted in up-regulation of ERalpha and down-regulation of ERbeta. With estrogen and tibolone treatment, ERbeta showed a significant increase but ERalpha showed no significant difference compared with Ovx.</p><p><b>CONCLUSIONS</b>Estrogen deficiency deteriorates bladder ultra structure and histology. Supplementary estrogen can improve bladder function which may be due to inhibition of collagen hyperplasia and increased SM density. ERbeta has an important role in mediating estrogen function in the bladder. Tibolone has a mild estrogenic action and has an effect on bladder function and structure to some degree.</p>