ObjectiveTo construct a multifunctional liposomal delivery system by replacing cholesterol（Chol） in conventional liposomes with saikosaponin D（SSD） and modifying with poloxamer 407（P407） for co-delivery of curcumin（Cur）. The system was evaluated for in vivo tumor targeting and inhibitory effects on mouse subcutaneous solid tumors. MethodsSingle-factor and orthogonal tests combined with information entropy weighting were used to optimize the formulation process of the liposome with encapsulation efficiency and absolute Zeta potential as indexes， and validation studies and liposomal characterization were performed. A subcutaneous solid tumor model was established by injecting H22 hepatocellular carcinoma cells subcutaneously into the dorsal surface of the right forelimb of mice. DiR-loaded traditional Chol liposomes（P407-DiR-Chol-LPs， PDCL） and novel SSD-based liposomes（P407-DiR-SSD-LPs， PDSL） were prepared by the optimized formulation process， and tail vein injection was performed to investigate the impact of SSD on liposome tumor targeting with small animal in vivo imaging. Mice were randomly divided into eight groups， including blank group， model group， free doxorubicin（DOX） group（2 mg·kg^-1）， free Cur group（8 mg·kg^-1）， free SSD group（10 mg·kg^-1）， P407-Cur-Chol-LPs（PCCL） group， P407-SSD-LPs（PSL） group， and P407-Cur-SSD-Lps（PCSL） group. Treatments were administered intraperitoneally every other day for seven doses. Antitumor efficacy and biocompatibility were evaluated by monitoring body weight change， organ indices， tumor volume and mass， relative tumor proliferation rate（T/C）， and tumor growth inhibition rate（TGI）. Histopathological analysis of liver， kidney， and tumor tissues was performed using hematoxylin-eosin（HE） staining. Serum levels of aspartate aminotransferase（AST）， alanine aminotransferase （ALT）， blood urea nitrogen（BUN）， and creatinine（Crea）in mice were quantified by fully automated biochemical analyzer. ResultsOrthogonal test yielded optimal ratios of Cur， SSD， and P407 to soybean phosphatidylcholine（SPC） as 1∶25， 1∶20， and 1∶4. The optimized PCSL exhibited spherical morphology with a particle size of 179.15 nm， a Zeta potential of -47.25 mV， and an encapsulation efficiency of 96.40%. Its in vitro release profile conformed to first-order kinetics， demonstrating excellent storage stability and hemocompatibility. In vivo imaging revealed that the fluorescence signal in tumor tissues and the fluorescence intensity ratio between tumors and organs were significantly higher in the PDSL group than in the PDCL group（P<0.05， P<0.01）. Among the treatment groups， PCSL group showed superior efficacy over free Cur group， free SSD group， PCCL group， and PSL group， with TGI>40% and T/C<60%， indicating pronounced anti-hepatocellular carcinoma effects（P<0.05， P<0.01）. Histopathology and serum biochemistry indicated minimal hepatorenal toxicity and improved hepatic and renal function in PCSL-treated mice. ConclusionReplacing Chol with SSD in preparing multifunctional drug delivery systems not only stabilizes liposomes but also yields superior anti-hepatocellular carcinoma efficacy， achieving the effect of drug-excipient integration. Co-delivery of Cur via this system can be used for treating subcutaneous solid tumors in hepatocellular carcinoma， providing new insights and technical approaches for anti-hepatocellular carcinoma research and the meridian-guiding and messenger-directing theory in traditional Chinese medicine.

The amino-terminal pro-C-type natriuretic peptide(NT-proCNP)is a diagnostic inflam-matory marker clinically used for diagnosing bacterial infections.This study aims to establish a phage display library of single-chain variable fragment(scFv)antibodies against canine NT-proC-NP and to screen for scFvs with high binding affinity to NT-proCNP.Initially,NT-proCNP was prepared using prokaryotic expression system and was used to immunize New Zealand White rab-bits.Upon achieving the desired serum titer,total RNA was extracted from the splenocytes of rab-bits and reverse transcribed into cDNA.Using this cDNA as a template,degenerate primers were employed to amplify the genes of the rabbit antibody light chain variable region(VL)and heavy chain variable region(VH).The VL and VH regions were spliced together to form a complete scFv fragment via overlap extension PCR.The scFv was then ligated into the phagemid pComb3XSS and electroporated into competent E.coli TG1 cells to construct a rabbit-derived anti-NT-proCNP scFv immunological library.This library underwent four rounds of enrichment and screening to isolate specific single-chain antibodies.The selected antibody was subsequently ex-pressed in a soluble form within a prokaryotic system,and its immunological activity was evalua-ted.Using phage display technology,this study successfully identified a single-chain antibody scFv-1-CNP with strong antigen-binding activity and genetic sequence characteristics of scFvs,providing a research direction for further exploration of scFv applications in the detection of NT-proCNP.

OBJECTIVE To construct an evaluation index system for the core competencies of ethics committee members of drug clinical trial institution,providing a basis for optimizing the training system for committee members,improving the quality of ethical review,and fully safeguarding the safety and rights of subjects.METHODS Using methods such as literature research and expert consultation,a preliminary core competency evaluation index system was constructed.The Delphi method was employed to revise and validate it,ultimately forming an evaluation index system for the core competencies of ethics committee members.Based on this system,a questionnaire survey was conducted among 90 ethics committee members from 29 drug clinical trial institutions nationwide,comparing their importance rating and self-assessment scores of the core competency indexes.RESULTS The evaluation system constructed included 4 primary indicators(ethics and professional knowledge,ethics review ability,communication and expression ability,moral integrity and work style)and 39 secondary indicators(familiarity with the content of clinical trial-related laws and regulations,ability to complete project ethics review and identify ethical defects in research protocols within a short period of time,ability to judge the scientific value of clinical research,etc.).The results of questionnaire survey showed that the interviewed ethics committee members had significant capability gaps in dimensions such as regulatory knowledge,ethical norms,review efficiency,risk judgment,and problem analysis.The differences between the importance rating scores of corresponding secondary indicators and the self-assessment scores were all no less than 0.38.CONCLUSIONS This study has developed a quantifiable and stratified core competency assessment tool for ethics committee members.It can provide a scientific framework for committee member training,qualification certification,and standardized management of ethics committees.

Objective:To investigate the distribution of CGG repeat numbers in the FMR1 gene among reproductive-age women in China, providing data reference for carrier screening and genetic counseling of Fragile X syndrome. Methods:This cross-sectional study recruited 16 610 reproductive-age women from 12 medical institutions between July 2022 and October 2023. Peripheral venous blood samples (3 mL) were collected, and genomic DNA was extracted. The number of CGG repeats in the FMR1 gene was determined using the triplet-primed polymerase chain reaction (TP-PCR) combined with capillary electrophoresis technology. Statistical analyses were performed to assess the prevalence and distribution of CGG repeat expansions. Results:Among 16 610 women of childbearing age, 5 684 (34.220%) women had the same number of CGG repeats in the two alleles of FMR1 gene, and 10 926 (65.780%) women had different numbers of repeats in the two alleles. Among the 33 220 FMR1 alleles in 16 610 women of reproductive age, the most common CGG repeat numbers were 29 [48.645% (16 160/33 220)] and 30 [26.276% (8 729/33 220)], while the most frequent CGG genotype was CGG 29/29 [24.726% (4 107/16 610)]. The CGG repeat numbers of FMR1 gene were normal in 16 498 women (99.326%). Among the 112 women (0.674%) with CGG repeat abnormities, 96 (0.578%) women were classified as intermediate carriers, 15 (0.090%) as premutation carriers, and 1 (0.006%) as a full mutation carrier, whose CGG genotype was (36, >200). Conclusion:In the general reproductive-age female population in China, the normal CGG repeat numbers of the FMR1 gene account for 99.326%, while the intermediate carrier rate is 0.578%, and the combined carrier rate of the premutation and full mutation types is 0.096%.

This study investigated the immunogenicity of type 5 replication deficient recombinant adenovirus comprising the fused receptor binding domain protein(RBD)and capsid protein(N)of the SARS-CoV-2 Omicron strain.The overlap PCR method was used to obtain the RBDgs3N fragment.The plasmid pKAd5ES-RBDgs3N was obtained through homologous recombination,and re-combinant virus was amplifiedand harvested after transfection of HEK293 cells.Cesium chloride density gradient centrifugation was used to purify the recombinant adenovirus,and the viral titer was determined through limited dilution analysis.The fusion protein ex-pression was identified with the western blot(WB)method.Recombinant adenovirus was injected intramuscularly into BALB/c mice,and the specific immune responses of the mice were detected with indirect ELISA and enzyme-linked immunosorbent spot technology.The titer of the recombinant adenovirus pKAd5ES-RBDgs3N virus seed was 1.168×1010 IU/mL.The RBDgs3N fusion protein was suc-cessfully expressed in HEK293 cells.Single dose intramuscular immunization significantly induced production of IgG antibodies against wild type(WT)SARS-CoV-2 virus and variant(Delta and Omicron)RBDs in mice,with antibody titers of 103.677 8,103.878 5,and 104.454 9,respectively.The specific antibody titer against N protein was 104.942 2.The level of IFN-γ secretion for RBD specific cellu-lar immunity was 1 452 SFC/106 cells.The type 5 replication deficient recombinant adenovirus with RBD and N gene fusion constructed in this study elicited high levels of specific antibodies against RBD and N protein and RBD specific cellular immunity in BALB/c mice,thus providing a basis for further evaluation of the recombinant adenovirus as a potential SARS-CoV-2 vaccine.

Hepatocellular carcinoma(HCC)is a cancer with extremely high mortality and incidence rates.In recent years,with the development of multidisciplinary approaches,key characteristics of cancer have gradually been revealed.Studies have shown that dysregulated energy metabolism and metabolic reprogramming in tumor cells form the basis for tumor cell growth and are critical factors in shaping the tumor microenvironment,immune evasion,and drug resistance.Non-coding RNAs(ncRNAs),as important regulatory factors,play a crucial role in the metabolic reprogramming of HCC.This article focuses on summarizing the mech-anisms by which ncRNAs regulate metabolic repro-gramming in HCC and explores their applications in clinical diagnosis,treatment,and prognostic assess-ment,aiming to provide new perspectives for effec-tive HCC treatment and the development of novel drug targets.

Objective:To analyze the iodine nutrition status and its related factors among adults aged 18 years and above in Zhejiang Province in 2022.Methods:A multistage stratified sampling method was used to select 4 320 adults aged 18 years and above from 16 on-site survey sites in Zhejiang Province for the study. A questionnaire was used to investigate the general demographic information and personal dietary characteristics of the study participants. Household edible salt and urine samples were collected to detect salt iodine content and urinary iodine level by using direct titration and cerium arsenate-catalyzed spectrophotometry, respectively, to evaluate the iodine nutritional status according to the standard. The multiple-ordered logistic regression model was used to analyze the factors influencing the urinary iodine concentration.Results:The age of the 4 320 study participants was (51.19±15.33) years, with males accounting for 44.44% (1 920). About 40.16% of adults (1 735) were from coastal areas and 56.37% (2 435) from urban areas. The salt iodine content, M ( Q1, Q3), of the 4 320 household edible salt samples was 21.10 (0.00, 24.16) mg/kg, including 1 662 non-iodized salt samples, 182 unqualified iodized salt samples and 2 476 qualified iodized salt samples. The rate of iodized salt coverage was 61.53%, and the rate of qualified iodized salt consumption was 57.31%. There was a statistically significant difference in the proportion of qualified iodized salt in adult households among different regions ( P<0.001), with the proportion of non-iodized salt gradually decreasing from coastal to inland areas ( χ 2trend=618.458, P<0.001). The urinary iodine concentration M ( Q1, Q3) was 137.60 (86.85, 210.60) μg/L in 4 320 adult urine samples, with the urinary iodine levels of<100, 100-199, 200-299, and≥300 μg/L accounting for 31.64% (1 367), 40.56% (1 752), 17.66% (763), and 10.14% (438), respectively. There was a nonlinear positive correlation between household salt iodine content and urinary iodine level in adults aged 18 years and above by using the χ 2 test for trend ( χ 2regression=231.10, P<0.001 and χ 2skew=28.81, P<0.001). Urinary iodine concentrations were higher in men than in women ( P=0.029) and higher in adults in rural areas than in urban areas ( P<0.001). There were statistically significant differences in the distribution of iodine nutritional status among adults of different ages, regions, and urban and rural areas (all P<0.001). The proportion of those with urinary iodine levels<100 μg/L gradually increased with age ( χ 2trend=37.493, P<0.001), and gradually decreased from coastal areas to inland areas ( χ 2trend=71.381, P<0.001). The results of the multiple-ordered logistic regression model analysis showed that compared with adults aged 18 to 44 years and male adults, those aged 45 to 59 years and female adults had lower urinary iodine levels, with OR (95% CI) of 0.75 (0.68-0.83) and 0.85 (0.76-0.95), respectively. Compared with adults in coastal and urban adults, those in sub-coastal, inland and rural adults had higher levels of urinary iodine, with OR (95% CI) of 1.89 (1.63-2.19), 2.02 (1.72-2.37) and 1.46 (1.28-1.66), respectively. Conclusion:The overall iodine nutrition level of adults aged 18 years and above in Zhejiang Province in 2022 is generally appropriate. However, there is a potential risk of iodine deficiency among adults in coastal areas.

Objective:To investigate the iodine nutrition status of pregnant women in Zhejiang Province, explore the impact of salt industry system reform on iodine nutrition level of pregnant women, and provide scientific basis for prevention and treatment of iodine deficiency disorders.Methods:From 2016 to 2021, a multi-stage stratified sampling method was used to select 100 pregnant women from 90 counties (cities, districts) in Zhejiang Province each year as survey subjects. Salt samples were collected from pregnant women's families, and one random urine sample was taken for salt iodine and urinary iodine level testing, respectively.Results:A total of 56 581 samples of household edible salt were collected from pregnant women, with a median salt iodine level of 23.20 mg/kg. Among them, 7 961 were non iodized salt, 45 803 were qualified iodized salt, and 2 817 were unqualified iodized salt. The iodized salt coverage rate was 85.93% (48 620/56 581), and the qualified iodized salt consumption rate was 80.95% (45 803/56 581). The proportion of non iodized salt increased from 10.05% (897/8 928) in 2016 to 15.09% (1 461/9 679) in 2021 (χ 2trend = 95.16, P < 0.001). A total of 56 581 urine samples were collected from pregnant women, with a median urinary iodine level of 130.50 μg/L. Among them, the proportions of urinary iodine levels < 150, 150 - 249, 250 - 499, and ≥500 μg/L were 58.32% (32 996/56 581), 27.24% (15 410/56 581), 12.24% (6 926/56 581), and 2.21% (1 249/56 581), respectively. The median urinary iodine level of pregnant women in inland areas was significantly higher than that in coastal areas ( Z = 19.15, P < 0.001). Furthermore, urinary iodine levels exhibited a non-linear decline as age increased (χ 2regression = 12.65, P < 0.001; χ 2partial = 22.65, P < 0.001) and as pregnancy progressed (χ 2regression = 37.28, P < 0.001; χ 2partial = 18.89, P < 0.001). Conclusions:The overall iodine nutrition status of pregnant women in Zhejiang Province is in a state of iodine deficiency (< 150 μg/L), and there is a greater risk in coastal areas compared to inland areas. However, in the context of the reform of the salt industry system, it is still necessary to strengthen the quality supervision of iodized salt, provide scientific iodine supplementation education, promote specialized iodized salt for pregnant women, and strengthen interventions for prevention and control of iodine deficiency disorders.

This study investigated the immunogenicity of type 5 replication deficient recombinant adenovirus comprising the fused receptor binding domain protein(RBD)and capsid protein(N)of the SARS-CoV-2 Omicron strain.The overlap PCR method was used to obtain the RBDgs3N fragment.The plasmid pKAd5ES-RBDgs3N was obtained through homologous recombination,and re-combinant virus was amplifiedand harvested after transfection of HEK293 cells.Cesium chloride density gradient centrifugation was used to purify the recombinant adenovirus,and the viral titer was determined through limited dilution analysis.The fusion protein ex-pression was identified with the western blot(WB)method.Recombinant adenovirus was injected intramuscularly into BALB/c mice,and the specific immune responses of the mice were detected with indirect ELISA and enzyme-linked immunosorbent spot technology.The titer of the recombinant adenovirus pKAd5ES-RBDgs3N virus seed was 1.168×1010 IU/mL.The RBDgs3N fusion protein was suc-cessfully expressed in HEK293 cells.Single dose intramuscular immunization significantly induced production of IgG antibodies against wild type(WT)SARS-CoV-2 virus and variant(Delta and Omicron)RBDs in mice,with antibody titers of 103.677 8,103.878 5,and 104.454 9,respectively.The specific antibody titer against N protein was 104.942 2.The level of IFN-γ secretion for RBD specific cellu-lar immunity was 1 452 SFC/106 cells.The type 5 replication deficient recombinant adenovirus with RBD and N gene fusion constructed in this study elicited high levels of specific antibodies against RBD and N protein and RBD specific cellular immunity in BALB/c mice,thus providing a basis for further evaluation of the recombinant adenovirus as a potential SARS-CoV-2 vaccine.

To systematically review randomized controlled trials （RCTs） of traditional Chinese medicine （TCM） intervention in ulcerative colitis （UC）， and analyze the characteristics of these studies and their outcome indicators， thereby providing references for the design of future RCTs of TCM intervention in UC and offering evidence supporting the clinical application of TCM in UC. A computerized search was conducted in the China National Knowledge Infrastructure （CNKI）， Wanfang Data， VIP， SinoMed， PubMed， Cochrane Library， EMbase， and Web of Science databases for RCTs of TCM intervention in UC published from January 2021 to August 2024. The risk of bias was assessed， and outcome indicators were qualitatively analyzed. A total of 555 RCTs were included， with a sample size of 44 853 participants. The largest sample size was 218 cases， and the smallest was 28 cases， with most studies focusing on 60-100 participants. Of the 386 RCTs that explicitly reported TCM syndrome types， the top three were large intestine dampness-heat syndrome （31.05%）， spleen and kidney yang deficiency syndrome （12.47%）， and spleen deficiency with dampness syndrome （9.17%）. The interventions， ranked by frequency of use， included internal Chinese medicine compounds/preparations （64.5%）， Chinese medicine compounds/preparations with retained enema （18.2%）， internal Chinese medicine compounds/preparations + external TCM treatment （5.95%）， and external TCM treatment alone （4.86%）. The treatment duration was mainly 4-8 weeks （64.86%）， with 61 studies （10.99%） reporting follow-up time. A total of 157 outcome indicators were used， with a frequency of 3 460 occurrences， classified into six domains： TCM syndromes and symptoms （346 occurrences， 10%）， symptoms/signs （541 occurrences， 15.64%）， physical and chemical examinations （2 119 occurrences， 61.24%）， quality of life （107 occurrences， 3.09%）， long-term prognosis （61 occurrences， 1.76%）， and safety events （284 occurrences， 8.21%）. The analysis reveals several limitations in the outcome indicators of TCM intervention in UC， including the lack of a basis for sample size calculation， non-standardized TCM syndrome classification， absence of trial design and registration， inadequate blinding and allocation concealment， adherence issues with interventions， imbalanced selection of surrogate and endpoint indicators， inconsistency in the timing of outcome measurements， design issues that require standardization， and ethical and safety concerns. It is recommended that future studies actively construct a set of core indicators for UC that include standardized TCM syndrome classification， clear efficacy evaluation indicators， key endpoint indicators， and reasonable measurement time points. Long-term prognostic impacts， comprehensive assessments of patients' quality of life， and consideration of economic benefits should be emphasized， providing a basis for the clinical practice of TCM in the treatment of UC.