Objective:To investigate immunotherapy effects and mechanism of BCG and recombinant BCG(rBCG)with c-di-AMP as adjuvant on melanoma in mice model.Methods:Melanoma mice model was established by B16F10 cell subcutaneous injec-tion in groin,and treated with 1×106 CFU of BCG and rBCG by adjacent injection of subcutaneous tumor for 3 times,respectively.Survival of melanotic mice,tumor growth and metastasis were observed.Tumor tissues of mice were isolated to prepare cell suspen-sion,and proportion of immune cells were detected by flow cytometry.Transcriptional levels of immune-related genes in tumor tissues were detected by qRT-PCR.Results:Both BCG and rBCG immunotherapy could significantly inhibit growth in melanoma mice and prolong survival time of mice.rBCG showed better inhibition on metastasis than BCG.Both strains significantly reduced proportion of M2-type macrophages and myeloid-derived suppressor cell associated with tumor growth and metastasis.Both two strains promoted infiltration of lymphocytes in tumor tissues,and rBCG significantly increased proportion of B cells in tumor.BCG immunotherapy upregulated transcription levels of metastasis-related cytokines,while rBCG therapy had no effects on transcriptions of these genes.Conclusion:Both BCG and rBCG have immunotherapeutic effects on melanotic mice,and rBCG with c-di-AMP as adjuvant shows better inhibition on tumor metastasis than BCG,which mechanism was related to regulation of immune response in tumor tissues.

Objective:To summarize domestic and foreign transitional readiness assessment tools for adolescents with chronic diseases and analyze the current status of their application in the clinic, so as to provide references for healthcare professionals to select appropriate tools.Methods:China National Knowledge Infrastructure, Wanfang Data, VIP and China Biomedical Literature Database, PubMed, Web of Science, Embase, Medline, CINAHL, Cochrane Library and scholar.google.com were searched for literature related to the assessment tools of transition readiness in adolescents with chronic illnesses from inception to March 29, 2024. Two researchers independently screened the literature and extracted the data.Results:A total of 491 articles were initially retrieved and 24 were included, involving a total of 8 transitional readiness assessment tools for adolescents with chronic diseases, including the Transition Readiness Assessment questionnaire, the University of North Carolina TRxANSITION Scale, Am I ON TRAC for adult care? Questionnaire, Transition Questionnaire, Self-Management and Transition to Adulthood with Rx=Treatment Questionnaire, Good 2 Go Questionnaire, State Assessment Questionnaire for Transition, and Self-assessment Scale of Transition Readiness for Adolescents. The Transition Readiness Assessment questionnaire and Self-Management and Transition to Adulthood with Rx=Treatment Questionnaire have good reliability and validity, comprehensive evaluation, wide applicability, simple use, and are suitable for clinical use.Conclusions:The quality of existing tools for assessing transitional readiness of adolescents with chronic diseases is mixed. The Transition Readiness Assessment questionnaire and Self-Management and Transition to Adulthood with Rx=Treatment Questionnaire are more appropriate options in the clinic, but they still need to be improved. Future studies need to continue to introduce tools to assess transitional readiness for chronic diseases in adolescents and develop localized assessment tools.

Objective:To investigate immunotherapy effects and mechanism of BCG and recombinant BCG(rBCG)with c-di-AMP as adjuvant on melanoma in mice model.Methods:Melanoma mice model was established by B16F10 cell subcutaneous injec-tion in groin,and treated with 1×106 CFU of BCG and rBCG by adjacent injection of subcutaneous tumor for 3 times,respectively.Survival of melanotic mice,tumor growth and metastasis were observed.Tumor tissues of mice were isolated to prepare cell suspen-sion,and proportion of immune cells were detected by flow cytometry.Transcriptional levels of immune-related genes in tumor tissues were detected by qRT-PCR.Results:Both BCG and rBCG immunotherapy could significantly inhibit growth in melanoma mice and prolong survival time of mice.rBCG showed better inhibition on metastasis than BCG.Both strains significantly reduced proportion of M2-type macrophages and myeloid-derived suppressor cell associated with tumor growth and metastasis.Both two strains promoted infiltration of lymphocytes in tumor tissues,and rBCG significantly increased proportion of B cells in tumor.BCG immunotherapy upregulated transcription levels of metastasis-related cytokines,while rBCG therapy had no effects on transcriptions of these genes.Conclusion:Both BCG and rBCG have immunotherapeutic effects on melanotic mice,and rBCG with c-di-AMP as adjuvant shows better inhibition on tumor metastasis than BCG,which mechanism was related to regulation of immune response in tumor tissues.

Objective:To summarize domestic and foreign transitional readiness assessment tools for adolescents with chronic diseases and analyze the current status of their application in the clinic, so as to provide references for healthcare professionals to select appropriate tools.Methods:China National Knowledge Infrastructure, Wanfang Data, VIP and China Biomedical Literature Database, PubMed, Web of Science, Embase, Medline, CINAHL, Cochrane Library and scholar.google.com were searched for literature related to the assessment tools of transition readiness in adolescents with chronic illnesses from inception to March 29, 2024. Two researchers independently screened the literature and extracted the data.Results:A total of 491 articles were initially retrieved and 24 were included, involving a total of 8 transitional readiness assessment tools for adolescents with chronic diseases, including the Transition Readiness Assessment questionnaire, the University of North Carolina TRxANSITION Scale, Am I ON TRAC for adult care? Questionnaire, Transition Questionnaire, Self-Management and Transition to Adulthood with Rx=Treatment Questionnaire, Good 2 Go Questionnaire, State Assessment Questionnaire for Transition, and Self-assessment Scale of Transition Readiness for Adolescents. The Transition Readiness Assessment questionnaire and Self-Management and Transition to Adulthood with Rx=Treatment Questionnaire have good reliability and validity, comprehensive evaluation, wide applicability, simple use, and are suitable for clinical use.Conclusions:The quality of existing tools for assessing transitional readiness of adolescents with chronic diseases is mixed. The Transition Readiness Assessment questionnaire and Self-Management and Transition to Adulthood with Rx=Treatment Questionnaire are more appropriate options in the clinic, but they still need to be improved. Future studies need to continue to introduce tools to assess transitional readiness for chronic diseases in adolescents and develop localized assessment tools.

In this study,the immunological characteristics of the PhoP protein were explored with a two-component system of Mycobacterium tuberculosis(Mtb).Bioinformatics was used to predict the B and T cell epitopes of the PhoP protein.A re-combinant expression plasmid was constructed by PCR analysis of the phoP sequence and cloning into the prokaryotic expres-sion vector pET-28a(+).Competent Escherichia coli BL21 cells were transformed with the recombinant plasmid and expres-sion was induced with IPTG.The recombinant PhoP protein was purified by affinity chromatography.Serum levels of PhoP-specific antibodies in Mtb-infected mice and tuberculosis(TB)patients were analyzed with an ELISA.BALB/c mice were im-munized with the PhoP recombinant protein by intramuscular injection.Sera of mice were collected and antibody titers were detected with an ELISA and specificity was assessed by West-ern blot analysis.Mouse splenocytes were isolated and the pro-portions of IFN-y-positive cells and cytokine levels were detec-ted with an ELISpot and ELISA,respectively.Bioinformatics i-dentified 24 B cell and 11 T cell epitopes of the PhoP protein.A prokaryotic recombinant vector of PhoP was successfully con-structed and the recombinant PhoP protein was obtained by purification.Specific antibody levels to PhoP in sera of Mtb in-fected mice and TB patients increased significantly,with preci-sion of 99.9％and 82.5％,and specificity of 100％,respectively.PhoP protein immunization successfully induced production of specific antibodies in mice.Stimulated by antigens in vitro,IL-2 and IFN-γ levels were significantly increased in the splenocytes of immunized mice.Immunization with the PhoP protein induce a humoral immune response and Thl-dominated cellular immu-nity,indicating that the PhoP protein was immunogenic with diagnostic efficacy for TB.These results lay a foundation to clari-fy the role of PhoP in Mtb infection and application for diagnosis and prevention of TB.

Objective To establish a HPLC method and fingerprints study for the simultaneous determination of loganic acid,gentiopicroside,paeoniflorin,vitexin,liquiritin,and ammonium glycyrrhizinatein,and other components in the alcohol extract of Mongolian medicine Qinggan Manu-4.Methods The Agilent Eclipse XDB-C18 column(250 mm×4.6 mm,5 μm)was used,with the detection wavelength of 230 nm for paeoniflorin,vitexin and liquiritin,237 nm for loganic acid,250 nm for glycyrrhizic acid,and 270 nm for gentiopicroside.The mobile phase was 0.1%phosphoric acid aqueous solution-acetonitrile with gradient elution.The flow rate was 1.0 mL/min,the column temperature was 30℃,and the injection volume was 10 μL.Results The mass concentrations of loganic acid,gentiopicroside,paeoniflorin,vitexin,liquiritin and ammonium gentiopicroside in the alcohol extract of Mongolian medicine Qinggan Manu-4 showed good linear relationships with the chromatographic peak area in the range of 0.020 3-0.121 0,0.053 3-0.317 7,0.021 3-0.127 0,0.011 5-0.069 0,0.014 1-0.083 8 and 0.035 1-0.209 1 mg/mL,respectively(r≥0.999 8).The average recovery rates of the six components were 94.75%,99.49%,92.32%,95.82%,101.29%and 98.04%,with the RSDs of 1.11%,0.76%,0.99%,2.75%,1.09%and 2.43%,respectively(n=6).The HPLC fingerprint of the alcoholic extract of Mongolian medicine Qinggan Manu-4 was established,using the chromatogram of sample S1 as the reference chromatogram,the control fingerprints were generated through multi-point correction and full-spectrum peak matching.A total of ten common peaks were identified,and after comparison with the reference substance,eight components were identified.Conclusion The established method and fingerprints are accurate,reliable,reproducible and specific,which provide a basis for the quality control and subsequent development of Mongolian medicine Qinggan Manu-4.

Objective:To observe the effects of modified Taohong Siwu Decoction on peripheral circulation status of affected limbs and serum thromboxane B 2 (TXB 2) and prostacyclin (PGI 2) levels in patients with traumatic limb fractures after open reduction and internal fixation. Methods:Randomized controlled trial study was conducted. 70 patients with traumatic limb fractures who underwent open reduction and internal fixation in the hospital from February 2020 to February 2023 were set as observation subjects and divided into control group (34 cases) and observation group (36 cases) according to the order of hospitalization. The control group was given conventional method for treatment, and the observation group was given modified Taohong Siwu Decoction adjuvant therapy on the basis of the control group. The treatment for both groups lasted for 2 weeks. TCM symptoms scores were evaluated before and after treatment. Ankle brachial index (ABI) was detected using a Doppler blood flow detector. Color Doppler ultrasound was used to detect the inner diameter, blood flow, and deep veins of the affected limb; serum levels of TXB 2 and PGI 2 were detected by ELISA. Results:After treatment, the total effective rate of clinical efficacy was 91.67% (33/36) in observation group and 73.53% (25/34) in control group, with statistical significance ( P<0.05). After treatment, the observation group had lower scores for qi and blood stasis (2.13 ± 0.43 vs. 3.61 ± 0.96, t=5.85), tendon and bone injury (1.62 ± 0.41 vs. 2.74 ± 0.58, t=9.37), swelling and ecchymosis (1.15 ± 0.31 vs. 2.28 ± 0.52, t=1.12), and liver and kidney deficiency (1.52 ± 0.24 vs. 2.15 ± 0.36, t=8.66) compared to the control group ( P<0.001); after treatment, the ABI (0.83 ± 0.03 vs. 0.74 ± 0.02, t=14.68), vascular diameter [(0.48 ± 0.13) mm vs. (0.42 ± 0.11) mm, t=2.08], and blood flow velocity [(60.24 ± 15.46) cm/s vs. (52.15 ± 12.11) cm/s, t=2.41] in the observation group were higher than those in the control group ( P<0.01 or P<0.05). After treatment, the serum TXB 2 levels [(140.76 ± 16.64) ng/L vs. (168.39 ± 25.28) ng/L, t=5.37] and TXB 2/PGI 2 (6.67 ± 1.24 vs. 9.33 ± 1.69, t=7.54) in the observation group were lower than those in the control group ( P<0.01); the level of PGI 2 [(21.27 ± 2.24) ng/L vs. (18.71 ± 1.79) ng/L, t=5.26] was higher in the control group ( P<0.01). The incidence of deep vein thrombosis after treatment in the observation group was 19.44% (7/36), while in the control group it was 44.12% (15/34), with statistical significance ( χ 2=4.94, P=0.026). Conclusion:Modified Taohong Siwu Decoction adjuvant therapy can enhance the clinical efficacy of patients with traumatic limb fractures after open reduction and internal fixation, relieve the TCM symptoms, improve the peripheral circulation status of affected limbs, reduce the postoperative hypercoagulability and prevent the deep venous thrombosis formation.

This study was aimed at understanding the genome-wide characterization and variations for three imported novel coronavirus(SARS-CoV-2)strains from different sources in the same period in Jinan at the viral genome level,to provide a sci-entific basis for further improving the prevention and control of COVID-19 outbreaks at Jinan port.We selected nasal and pha-ryngeal swab samples from three cases of imported asymptomatic COVID-19infectionat Jinan port;performed second-genera-tion whole-genome sequencing;and analyzed the variant loci and homology withvarious bioinformatics software.The whole ge-nome sequence of SARS-CoV-2 was successfully obtained from three samples of asymptomatic infected cases,and had full lengths ranging from 29 835 bp to 29 844 bp.Pangolin typing results indicated that the genotypes of the three samples were O-micron BQ.1,BQ.1.1,and BQ.1.12.Compared with the original Wuhan strain,the three samples produced mutations at 77,80,and 78 base sites,respectively,involving 60-63 non-synonymous mutations,mainly in the S and ORF1ab genes.Omicron BQ.1 is an imported variant of the SARS-CoV-2 virus and was detected for the first time at Jinan port.From a molecular biolo-gy perspective,this study provides a theoretical basis for the source tracing and prevention and control of COVID-19 at theport.

Objective: To compare the differences in the stress distribution in simulated first molars prepared with three rotary nickel-titanium instruments. Methods : Four simulated first molars were prepared without instruments and with Reciproc, WaveOne and Protaper. Before and after preparation, each simulated molar was scanned by Micro-CT. The data were imported to Mimics 16.0 software to establish three-dimensional models. Finite element analysis was processed with Abaqus 6.14 software under conditions of longitudinal and lateral load. Results: Under vertical load conditions, the maximum von Mises stress of the enamel increased by 1.36%, 21.48% and 20.99% in the Reciproc, WaveOne and Protaper groups, respectively, after preparation, while the maximum von Mises stress of the cementum increased by 55.98%, 41.18% and 33.04%, respectively, and the maximum von Mises stress of the alveolar bone increased by 45.55%, 40.37% and 24.09%, respectively. Under 45° lateral load conditions, the maximum von Mises stress of the enamel increased by 1.79%, -4.58% and 3.82% in the Reciproc, WaveOne and Protaper groups, respectively, after preparation, while the maximum von Mises stress of cementum increased by 16.33%, 7.58% and 4.32%, respectively, and the maximum von Mises stress of the alveolar bone increased by 46.82%, 36.40% and 8.29%, respectively. Under the same conditions, the von Mises stresses of the cementum and alveolar bones of the simulated molars were higher after preparation than before preparation, especially under lateral load conditions. The stress was focused on the border between the crown and the root. The von Mises stress of the cementum and alveolar bones increased much more in the Reciproc group than in the other two groups under both conditions. Conclusion The von Mises stress of simulated molars was greater after preparation than before preparation. The von Mises stress of the cementum and alveolar bones increased much more in the Reciproc group than in the other two groups.