A robust, selective and highly sensitive chemiluminescent （CL） platform for protein assay was presented in this paper. This novel CL approach utilized rolling circle amplification （RCA） as a signal enhancement technique and the 96-well plate as the immobilization and separation carrier. Typically, the antibody immobilized on the surface of 96-well plate was sandwiched with the protein target and the aptamer-primer sequence. This aptamer-primer sequence was then employed as the primer of RCA. Based on this design, a number of the biotinylated probes and streptavidin-horseradish peroxidase （SA-HRP） were captured on the plate, and the CL signal was amplified. In summary, our results demonstrated a robust biosensor with a detection limit of 10 fM that is easy to be established and utilized, and devoid of light source. Therefore, this new technique .will broaden the perspective for future development of DNA-based biosensors for the detection of other protein biomarkers related to clinical diseases, by taking advantages of high sensitivity and selectivity.

A robust,selective and highly sensitive chemiluminescent (CL) platform for protein assay was presented in this paper.This novel CL approach utilized rolling circle amplification (RCA) as a signal enhancement technique and the 96-well plate as the immobilization and separation carrier.Typically,the antibody immobilized on the surface of 96-well plate was sandwiched with the protein target and the aptamer-primer sequence.This aptamer-primer sequence was then employed as the primer of RCA.Based on this design,a number of the biotinylated probes and streptavidin-horseradish peroxidase (SA-HRP) were captured on the plate,and the CL signal was amplified.In summary,our results demonstrated a robust biosensor with a detection limit of 10 fM that is easy to be established and utilized,and devoid of light source.Therefore,this new technique will broaden the perspective for future development of DNA-based biosensors for the detection of other protein biomarkers related to clinical diseases,by taking advantages of high sensitivity and selectivity.

AIM: To analyze the electronic product use time, writing time, playing piano time and outdoor activity time and the distribution of myopia in 586 cases of school age children in our hospital. ·METHODS: A retrospective analysis of 586 cases of children aged 6 to 12 years old in the outpatient department was established. Personalized files were used to record the uncorrected visual acuity, optometry, slit lamp, fundus mirror and strabismus. The cumulative use time of electronic products ( including computer, mobile phone, iPad) , writing time, whether to play the piano and outdoor activities time with the eye situation were recorded. Statistical analysis of the age group of myopia, the cumulative use of electronic age in different age groups, writing time, whether playing piano and outdoor activities and the distribution of myopia occurred. · RESULTS: ( 1 ) With the increasing of age, the distribution of uncorrected eyesight was in children mostly mild myopia, and the proportion of mild myopia was significantly higher than that of moderate and high myopia. ( 2 ) Electronic products use time distribution:the proportion of playing electronic products ( including mobilephone, computers, iPad) accounted for 76. 8%, of which 9 years old, 10 years old the cumulative use of electronic products with a long time was higher than other age groups. ( 3 ) Distribution of writing time: the proportion of write homework ≤1h was significantly lower than the proportion of writing homework > 1h (37. 2% vs 62. 8%), of which 9 and 10 years old children cumulative write time was higher than other age group. ( 4 ) Distribution of playing the piano: the proportion of playing piano time less than 1h was significantly higher than the proportion of playing piano time more than 1h ( 89. 1% vs 10. 9%). ( 5 ) Distribution of outdoor activities: the proportion of outdoor activities ≤1h in children at school age was significantly higher than that of outdoor activities > 1h (91. 8% vs 8. 9%). ·CONCLUSION: With the age exposure to electronic products becoming younger, heavy learning tasks and less outdoor activities, myopia occurred in advance of age. So health examination and eye guidance, reducing the amount of work appropriately, increasing outdoor activities will slow the development of early childhood myopia.

OBJECTIVETo explore the effects of exogenous carbon monoxide-releasing molecules 2 (CORM-2) on the vitality and toxicity of E. coli ATCC 25922, and to analyze the potential mechanism.

METHODS(1) In vitro experiments. Standard strains of E. coli ATCC 25922 were divided into groups A (without addition), B, C, D, and E according to the random number table, and then the latter 4 groups were respectively cultured with 1.2 mmol/L CORM-2, 1.6 mmol/L CORM-2, 1.2 mmol/L inactive CORM-2 (iCORM-2), 1.6 mmol/L iCORM-2, with six samples in each group. After being cultured for 0, 3, 5, 8, 10, 12, 16, 20, 24, 27, 30, 48 hours, proliferative vitality of E. coli was examined (denoted as absorption value under 600 nm wavelength), and bacteria number was counted. Other standard strains of E. coli ATCC 25922 were divided into groups F (without addition) and G (cultured with 0.8 mmol/L CORM-2), the expressions of genes fliA, dnaK, marA, and waaQ related to E. coli were detected by quantitative real-time (qRT)-PCR. (2) In vivo experiments. Other standard strains of E. coli ATCC 25922 were grouped as A', B', C', D', and E' and treated with the same method as that in groups A, B, C, D, and E, and 0.5 mL bacterial liquid of each group were collected when the absorption value of bacterial liquid in group A' was equal to 0.4 (under 600 nm wavelength). Seventy-two C57BL/6 mice were divided into groups, namely blank control (without treatment), H, I, J, K, and L according to the random number table, with 12 mice in each group. The mice in the latter 5 groups were intraperitoneally injected with 0.5 mL bacterial suspension of groups A', B', C', D', and E' respectively. After injection, general condition of mice in groups H, I, J, K, and L was observed. The serum levels of TNF-α and IL-6 were determined at post injection hour (PIH) 6, 12. The liver and lung samples were harvested for determination of myeloperoxidase (MPO) activity at PIH 12. The same process was carried out in blank control group. Data were processed with repeated measure analysis of variance (ANOVA), factorial design ANOVA, one-way ANOVA, and t test.

RESULTS(1) In vitro experiments. Compared with those of groups A and D, the proliferative vitality and bacteria number of E. coli in group B were all decreased (with F values respectively 1170.80, 217.52, P values all below 0.01). Compared with those of groups A and E, the proliferative vitality and bacteria number of E. coli in group C were also obviously decreased (with F values respectively 7948.34, 14 432.85, P values all below 0.01). Compared with those in group F, the expression of fliA was downregulated, while the expressions of dnaK, marA, and waaQ were upregulated in group G (with t values 30.28, -165.54, -168.88, -187.28, P values all below 0.01). (2) In vivo experiments. Symptoms including listlessness and tachypnea were observed in mice in groups H, K, and L, and they were ameliorated or not obvious in groups I and J. At PIH 6, the serum levels of TNF-α and IL-6 in groups H and K were respectively (647.3 ± 3.8) pg/mL, (3.44 ± 0.22) ng/mL and (639.3 ± 0.8) pg/mL, (2.47 ± 0.32) ng/mL, which were obviously higher than those in group I [(124.6 ± 10.7) pg/mL, (1.03 ± 0.16) ng/mL, with t values from 15.22 to 84.03, P values all below 0.01]. The serum levels of TNF-α and IL-6 in group J at PIH 6, 12 were also obviously decreased as compared with those in groups H and L (with t values from 19.27 to 245.34, P values all below 0.01). MPO activity of liver and lung tissues were significantly attenuated in group I at PIH 12 as compared with those in groups H and K, and it was also attenuated in group J when compared with those in groups H and L (with t values respectively from 17.36 to 18.92 and 2.35 to 3.61, P values all below 0.01).

CONCLUSIONSCORM-2 can obviously inhibit the vitality and toxicity of E. coli, which might be attributable to regulation of expressions of genes fliA, dnaK, marA, and waaQ of E. coli.

Animals ; Carbon Monoxide ; metabolism ; DNA-Binding Proteins ; metabolism ; Escherichia coli ; drug effects ; metabolism ; physiology ; Escherichia coli Proteins ; metabolism ; Glycosyltransferases ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; Interleukin-6 ; blood ; Liver ; enzymology ; Lung ; enzymology ; Mice ; Mice, Inbred C57BL ; Organometallic Compounds ; pharmacology ; Peroxidase ; metabolism ; Sigma Factor ; metabolism ; Tumor Necrosis Factor-alpha ; blood

Objective To evaluate the effect of self -efficacy training on hemiplegic patients after ischemic stroke. Methods Sixty cases were randomly divided into control group (n =30 )and self -efficacy training group (n =30).The self -efficacy group received rehabilitation training according to the self -efficacy theory (5 times /week, 40 min /per time,1 0 weeks of intervention).The movement and balance function were evaluated using Fugl -Meyer score (FMA)and the activities of daily living were evaluated by the task analysis scale and Barthel index.The self -efficacy level and quality of life were evaluated by the general self -efficacy scale (GSES)and medical outcomes study (SF -36).Results The scores of FMA,Barthel,SF -36 and GSES had no significant differences between the two groups before training (P >0.05 )and all indexes improved after training.Six scores of Barthel and all scores of FMA,SF -36 and GSES of self -efficacy group were significantly higher than those of control group after training (all P <0.05 ).Conclusion Self -efficacy application could improve the motor function on the rehabilitation of hemiplegic patients after ischemic stroke.

Objective To explore the efficiency and safety of cetuximab combined with radiotherapy on patients with advanced nasopharyngeal carcinoma.Methods Forty patients with advanced nasopharyngeal car-cinoma were divided into control group and treatment group , each group 20 cases.Control group received regular radiotherapy and chemotherapy , the treatment group received cetuximab on the basis of control group.Af-ter the therapy , the efficiency and incidence of adverse drug reactions and keep record on the relapse free survival rate in the following 3 years.Results The response rate in treatment group was 55.0%, significantly higher than that in control group , which was 30.0%( P <0.05 ).The levels of CD4 +and CD8 +had no significant difference between the two groups ( P >0.05 ).The incidence of dermatitis , gastrointestinal reac-tions, hypersensitivity had no significant difference of the two groups ( P>0.05);and the incidence of oral mucositis in treatment group was higher than that of control group ( P<0.05 ).The survival rate in the sec-ond and third year in treatment group was significant higher than that in control group ( P<0.05 ) , but the one year survival rate had no signifi-cant difference ( P >0.05 ).Conclusion Cetuximab combined with radiotherapy and chemotherapy was an efficient therapy.

Objective:Based on the forgetting curve principle to construct a model for the popularization of cardiopulmonary resuscitation among college students and validate its effectiveness, providing reference for the promotion of cardiopulmonary resuscitation.Methods:A randomized controlled trial was conducted. In October 2021, a multistage random sampling method was used to select 262 freshmen and sophomores from the School of Mathematics and Physics at Leshan Normal University as the control group, and 262 freshmen and sophomores from the School of Electronic Information and Artificial Intelligence as the experimental group.The control group received conventional training, while the experimental group, in addition to conventional training, received cardiopulmonary resuscitation retraining based on the Ebbinghaus forgetting curve at three time intervals: within 1 week, within 1 month, and within 1 to 6 months after training. The retraining included online self-learning, on-site observation reinforcement, and social practice sublimation. The cardiopulmonary resuscitation knowledge awareness rate and cardiopulmonary resuscitation skill assessment of the two groups were compared before training, immediately after training, and 6 months after training.Results:The final number of participants was 256 in the control group (126 males and 130 females) and 224 in the experimental group (110 males and 114 females). There were no statistically significant differences in cardiopulmonary resuscitation knowledge awareness rate and cardiopulmonary resuscitation skill scores between the two groups before and immediately after training (both P>0.05). Immediately after training, the average cardiopulmonary resuscitation knowledge awareness rate in the experimental and control groups were 91% (1 626/1 792) and 88% (1 806/2 048), respectively, which were significantly higher than the pre-training rates of 55% (989/1 792) and 55% (1 129/2 048) ( χ2=8.46, 7.30, both P<0.01). The cardiopulmonary resuscitation skill scores were (42.00 ± 3.55) and (41.68 ± 3.40), respectively, higher than the pre-training scores of (15.41 ± 4.85) and (15.92 ± 4.66) ( t=84.47, 45.56, both P<0.05). Six months after training, the awareness rates of determination of consciousness and cardiac arrest judgment, cardiopulmonary resuscitation steps, chest compression location, frequency, depth, and the ratio of chest compressions to artificial ventilation in the experimental group in college students were 90% (201/224), 93% (208/224), 92% (206/224), 93% (208/224), 89% (200/224), and 94% (210/224), respectively, higher than those in the control group, which were 62% (158/256), 71% (182/256), 77% (198/256), 73% (186/256), 70% (178/256), and 69% (176/256) ( χ2 values were 4.21-7.03, all P<0.01). The cardiopulmonary resuscitation skill score in the experimental group was (49.77 ± 3.68), higher than the control group′s (34.95 ± 4.79) ( t=38.25, P<0.01). The proportion of students confident in using cardiopulmonary resuscitation to save others was 92% (206/224) in the experimental group, higher than 58% (148/256) in the control group ( χ2=7.68, P<0.01). Six months after training, the cardiopulmonary resuscitation skill score in the experimental group was (49.77 ± 3.68), higher than the immediate post-training score of (42.00 ± 3.55), whereas the control group's cardiopulmonary resuscitation skill score was (34.95 ± 4.79), lower than the immediate post-training score of (41.68 ± 3.40) ( t=22.74, 18.33, both P<0.01). Conclusions:The cardiopulmonary resuscitation popularization model for college students based on the forgetting curve is conducive to mastering cardiopulmonary resuscitation knowledge and skills among trained college students, enhancing their confidence in using cardiopulmonary resuscitation techniques to rescue others. Moreover, it helps save the human, financial, and material resources required for cardiopulmonary resuscitation training to a certain extent and is worthy of promotion.

Purpose: Pancreatic cancer (PC) is a common malignant tumor of the digestive system, and its 5-year survival rate is only 4%. N6-methyladenosine (m6A) RNA methylation is the most common post-transcriptional modification and dynamically regulates cancer development, while its role in PC treatment remains unclear. Materials and Methods: We treated PC cells with gemcitabine and quantified the overall m6A level with m6A methylation quantification. Real-time quantitative reverse transcription polymerase chain reaction and Western blot analyses were used to detect expression changes of m6A regulators. We verified the m6A modification on the target genes through m6A-immunoprecipitation (IP), and further in vivo experiments and immunofluorescence (IF) assays were applied to verify regulation of gemcitabine on Wilms’ tumor 1–associated protein (WTAP) and MYC. Results: Gemcitabine inhibited the proliferation and migration of PC cells and reduced the overall level of m6A modification. Additionally, the expression of the “writer” WTAP was significantly downregulated after gemcitabine treatment. We knocked down WTAP in cells and found target gene MYC expression was significantly downregulated, m6A-IP also confirmed the m6A modification on MYC. Our experiments showed that m6A-MYC may be recognized by the “reader” IGF2BP1. In vivo experiments revealed gemcitabine inhibited the tumorigenic ability of PC cells. IF analysis also showed that gemcitabine inhibited the expression of WTAP and MYC, which displayed a significant trend of co-expression. Conclusion Our study confirmed that gemcitabine interferes with WTAP protein expression in PC, reduces m6A modification on MYC and RNA stability, thereby inhibiting the downstream pathway of MYC, and inhibits the progression of PC.

BACKGROUNDThe frequencies of regulatory T cells (Tregs) increased over the HIV infection but its counts actually decreased. We proposed that the decrease of Treg counts may cause the reduction of inhibitory effect and thereby account for the over-activation of Tregs during HIV infection. However, it remains unknown whether Tregs are also over-activated and thereafter the activation induced death may lead to the decrease of Tregs.

METHODSTregs were defined as CD4(+)CD25(+)CD127(lo/-) T cells. Eighty-one HIV-1 infected patients were enrolled in our study, and twenty-two HIV-1 seronegative donors were recruited as the control. The levels of HLA-DR on Tregs were determined by FACSAria flow cytometer.

RESULTSCompared to HIV-1 seronegative donors, the levels of HLA-DR on CD4(+)CD25(+)CD127(lo/-) Tregs were significantly increased in HIV-1 infected patients, and its increase was positively associated with viral loads (r = 0.3163, P = 0.004) and negatively with CD4 T-cell counts (r = -0.4153, P < 0.0001). In addition, significant associations between HLA-DR expression on CD4(+)CD25(+)CD127(lo/-) Tregs and the percentages of HLA-DR, CD38, Ki67 expressing CD4(+) and CD8(+) T cells were also identified.

CONCLUSIONHLA-DR on Tregs is a good marker for viral replication and disease progression. The over-activation of Tregs might result in the decrease of Tregs.

ADP-ribosyl Cyclase 1 ; metabolism ; Adult ; Aged ; CD4-Positive T-Lymphocytes ; immunology ; metabolism ; Cells, Cultured ; Female ; Flow Cytometry ; HIV Infections ; immunology ; metabolism ; HLA-DR Antigens ; metabolism ; Humans ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Interleukin-7 Receptor alpha Subunit ; metabolism ; Lymphocyte Activation ; immunology ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; immunology ; metabolism ; Young Adult

Objective To compare the cost-effectiveness of two anti-hypertensive therapy regimens,Compound anti-hypertensive tablets and other common anti-hypertensive agents,in the treatment program of Primary Hypertension.Methods We conducted a cost-effectiveness analysis based on a community trial.Two communities'primary hypertensive patients were enrolled to receive different therapy drugs:Compound anti-hypertensive tablets(Group A)or other common anti-hypertensive agents(Group B).Blood pressure,medicine used,and adverse drug reactions were observed and recorded for one year,and then costeffectiveness ratio of the two groups and incremental ratio were calculated.We considered a 30%drug price fluctuating load to make the sensitivity analysis.Results 2505 cases were enrolled with 1529 cases in group A and 976 cases in group B.The cost-effectiveness ratios were 418.1 and 1057.7 for Group A and B respectively while the incremental cost-effectiveness of Group B vs.Group A was 19 202.2.The results were insensitive to variation in the costs of drugs over clinically reasonable ranges.Conclusion Compound anti-hypertensive tablets appeared to be relatively cost-effective when compared to common drugs for the treatment of primary hypertension.