Objective:To construct a nanoparticle vaccine displaying the prefusion F (preF) protein of respiratory syncytial virus (RSV) using the I53-50 protein nanoparticle platform, and to systematically evaluate its immunogenicity and protective efficacy.Methods:The RSV preF trimer antigen was genetically fused to I53-50A and assembled in vitro with I53-50B to form preF-I53-50 nanoparticles, theoretically displaying 20 preF antigens per particle. The structure and purity were characterized by size-exclusion chromatography, SDS-PAGE, and negative-stain electron microscopy. BALB/c mice were intramuscularly immunized with varying doses (1 μg or 5 μg) of preF antigen or an equimolar amount of preF-I53-50 nanoparticles. Humoral immunity, B-cell responses, and protective efficacy were assessed following intranasal viral challenge.Results:The preF-I53-50 nanoparticles self-assembled into spherical structures (50-60 nm in diameter) with uniformly arrayed antigens. The nanoparticle vaccine enhanced RSV-specific IgG1 and IgG2a antibody responses, promoting a Th1-biased immune profile. At equimolar preF doses, the neutralizing antibody titers induced by 1 μg and 5 μg nanoparticle formulations were 2.8-fold and 2.3-fold higher, respectively, than those elicited by preF alone ( P<0.05). Notably, even the low-dose nanoparticle group outperformed the high-dose preF group (1.6-fold increase). Viral challenge experiments demonstrated that preF-I53-50 effectively suppressed pulmonary viral replication, mitigated pathological damage, and induced stronger germinal center and memory B-cell responses, suggesting enhanced B-cell affinity maturation and long-term immune memory. Conclusion:The preF-I53-50 vaccine improves the immunogenicity and protective efficacy of RSV preF through multivalent antigen display.

Objective To investigate the characteristics of changes in pain-anxiety-depression-fatigue symptom clusters and their possible associated factors in adolescents with acute lymphoblastic leukemia(ALL)during early chemotherapy.Methods A prospective longitudinal study was conducted from Nov 2019 to Oct 2021,enrolling newly diagnosed adolescent ALL patients from 5 tertiary or pediatric specialty hospitals in Shanghai,Zhejiang Province,Sichuan Province,Anhui Province and Guangdong Province.Patient-reported pain,anxiety,depression,and fatigue were collected at five time points within the first nine weeks of chemotherapy using the PROMIS Pediatric-25 instrument.Latent profile analysis(LPA)and latent transition analysis(LTA)were applied to explore the latent classes of symptom clusters,their transition probabilities over time,and possible risk or protective factors associated with class membership.Results A total of 134 ALL cases were enrolled,and symptom clusters at all the 5 time points(T1-T5)were consistently classified into three groups of mild,moderate and severe symptoms.The severe symptoms group accounted for the largest proportion at each time point(54.5%,59.7%,66.4%,49.3%,and 47.0%,respectively),while the mild and moderate symptoms groups showed an initial decline followed by an increase.Among participants,40.2%maintained the same symptom status,and 77.4%experienced at least one episode of severe symptom status during the trajectory.Religious affiliation(T5)and family monthly income>5 000 Yuan(T2,T4 and T5)served as protective factors against severe symptoms.Higher baseline fatigue(T1)was associated with membership in the severe symptoms group at subsequent time points.Conclusion Pain-anxiety-depression-fatigue symptoms in adolescents with ALL during early chemotherapy can be categorized into mild,moderate and severe symptoms with dynamic transitions over time.Higher baseline fatigue was associated with increased risk of severe symptoms,whereas higher family income and religious affiliation appeared protective effects.

Following the release of the Technical Requirements on Quality Control and Standard Establishment of Traditional Chinese Medicine Formula Granules by the National Medical Products Administration in 2021, Chinese Pharmacopoeia Commission has promulgated 296 national drug standards so far, and most provinces have started the work of establishing provincial standards as supplements. The promulgation of standards fostered high-quality development of the industry. Since the implementation of national and provincial standards for more than three years, enterprises have gained deep understanding and hands-on experiences on the characteristics, technical requirements, production process, and quality control of traditional Chinese medicine(TCM) formula granules. Meanwhile, challenges have emerged restricting the high-quality development of this industry, including how to formulate quality control strategies for medicinal materials and decoction pieces, how to reduce manufacturing costs, and how to improve the pass rate and product stability under high standards. Based on the work experiences from standard management and process research, this article analyzed the distribution of sources, processing methods, dry extract rate ranges, process requirements for volatile oil-containing decoction pieces, control measures of safety indices, characteristics and trends of setting characteristic chromatograms or fingerprints, characteristics and trends of setting content ranges, and main differences between national standards and provincial standards. On the one hand, this article aims to present main characteristics for deeply understanding different indicators in standards and provide basic ideas for establishing quality and process control systems. On the other hand, from the perspective of industrial practice, suggestions are put forward on the important aspects that need to be focused on in the quality and process control of TCM formula granules.
Drugs, Chinese Herbal/chemistry* ; Quality Control ; Medicine, Chinese Traditional/standards* ; China ; Drug Industry/standards*

Objective To investigate the characteristics of changes in pain-anxiety-depression-fatigue symptom clusters and their possible associated factors in adolescents with acute lymphoblastic leukemia(ALL)during early chemotherapy.Methods A prospective longitudinal study was conducted from Nov 2019 to Oct 2021,enrolling newly diagnosed adolescent ALL patients from 5 tertiary or pediatric specialty hospitals in Shanghai,Zhejiang Province,Sichuan Province,Anhui Province and Guangdong Province.Patient-reported pain,anxiety,depression,and fatigue were collected at five time points within the first nine weeks of chemotherapy using the PROMIS Pediatric-25 instrument.Latent profile analysis(LPA)and latent transition analysis(LTA)were applied to explore the latent classes of symptom clusters,their transition probabilities over time,and possible risk or protective factors associated with class membership.Results A total of 134 ALL cases were enrolled,and symptom clusters at all the 5 time points(T1-T5)were consistently classified into three groups of mild,moderate and severe symptoms.The severe symptoms group accounted for the largest proportion at each time point(54.5%,59.7%,66.4%,49.3%,and 47.0%,respectively),while the mild and moderate symptoms groups showed an initial decline followed by an increase.Among participants,40.2%maintained the same symptom status,and 77.4%experienced at least one episode of severe symptom status during the trajectory.Religious affiliation(T5)and family monthly income>5 000 Yuan(T2,T4 and T5)served as protective factors against severe symptoms.Higher baseline fatigue(T1)was associated with membership in the severe symptoms group at subsequent time points.Conclusion Pain-anxiety-depression-fatigue symptoms in adolescents with ALL during early chemotherapy can be categorized into mild,moderate and severe symptoms with dynamic transitions over time.Higher baseline fatigue was associated with increased risk of severe symptoms,whereas higher family income and religious affiliation appeared protective effects.

Objective:To construct a nanoparticle vaccine displaying the prefusion F (preF) protein of respiratory syncytial virus (RSV) using the I53-50 protein nanoparticle platform, and to systematically evaluate its immunogenicity and protective efficacy.Methods:The RSV preF trimer antigen was genetically fused to I53-50A and assembled in vitro with I53-50B to form preF-I53-50 nanoparticles, theoretically displaying 20 preF antigens per particle. The structure and purity were characterized by size-exclusion chromatography, SDS-PAGE, and negative-stain electron microscopy. BALB/c mice were intramuscularly immunized with varying doses (1 μg or 5 μg) of preF antigen or an equimolar amount of preF-I53-50 nanoparticles. Humoral immunity, B-cell responses, and protective efficacy were assessed following intranasal viral challenge.Results:The preF-I53-50 nanoparticles self-assembled into spherical structures (50-60 nm in diameter) with uniformly arrayed antigens. The nanoparticle vaccine enhanced RSV-specific IgG1 and IgG2a antibody responses, promoting a Th1-biased immune profile. At equimolar preF doses, the neutralizing antibody titers induced by 1 μg and 5 μg nanoparticle formulations were 2.8-fold and 2.3-fold higher, respectively, than those elicited by preF alone ( P<0.05). Notably, even the low-dose nanoparticle group outperformed the high-dose preF group (1.6-fold increase). Viral challenge experiments demonstrated that preF-I53-50 effectively suppressed pulmonary viral replication, mitigated pathological damage, and induced stronger germinal center and memory B-cell responses, suggesting enhanced B-cell affinity maturation and long-term immune memory. Conclusion:The preF-I53-50 vaccine improves the immunogenicity and protective efficacy of RSV preF through multivalent antigen display.

Objective:To establish a Plaque-reduction Neutralization Test (PRNT) for the detection of neutralizing antibody titers of Human Respiratory Syncytial Virus (HRSV) and optimize the conditions for preliminary application.Methods:The CHO expression system was used to produce palivizumab monoclonal antibody (palivizumab) and the influencing factors such as cell type, cell culture duration, fixation and permeabilization protocols, and blocking agents. The reproducibility of the method was verified and its correlation was verified with conventional PRNT. Finally, the optimized PRNT assay was further used to determine neutralizing antibody titers against HRSV subtypes A and B in BALB/c mouse serum (immunized by intramuscular injection of HRSV fusion proteins).Results:Palivizumab was expressed at approximately 50 mg/L. The optimal working conditions for PRNT were as follows: culturing HEp-2 cells for 2 days, fixing with 4% (V/V) paraformaldehyde at room temperature for 15 min followed by 0.2% (V/V) Triton X-100 permeabilization for 15 minutes as the optimal fixation-permeabilization and removing the blocking step. The overall coefficient of variation (CV) for the reproducibility validation of this method was <15%, showing a good linear relationship with the conventional PRNT. The Spearman correlation coefficient r s was 0.983. This method was used to detect neutralizing antibody titers in mouse sera against HRSV subtype A strain long and subtype B strain 9320, and the fusion proteins combined with AlOH and CpG adjuvant induced the highest neutralizing antibody titers in mice. Conclusion:The HRSV neutralizing antibody assay established in this study is rapid, reproducible, high-throughput, and can be used to detect neutralizing antibodies to HRSV subtypes A and B.

Objective:To establish a Plaque-reduction Neutralization Test (PRNT) for the detection of neutralizing antibody titers of Human Respiratory Syncytial Virus (HRSV) and optimize the conditions for preliminary application.Methods:The CHO expression system was used to produce palivizumab monoclonal antibody (palivizumab) and the influencing factors such as cell type, cell culture duration, fixation and permeabilization protocols, and blocking agents. The reproducibility of the method was verified and its correlation was verified with conventional PRNT. Finally, the optimized PRNT assay was further used to determine neutralizing antibody titers against HRSV subtypes A and B in BALB/c mouse serum (immunized by intramuscular injection of HRSV fusion proteins).Results:Palivizumab was expressed at approximately 50 mg/L. The optimal working conditions for PRNT were as follows: culturing HEp-2 cells for 2 days, fixing with 4% (V/V) paraformaldehyde at room temperature for 15 min followed by 0.2% (V/V) Triton X-100 permeabilization for 15 minutes as the optimal fixation-permeabilization and removing the blocking step. The overall coefficient of variation (CV) for the reproducibility validation of this method was <15%, showing a good linear relationship with the conventional PRNT. The Spearman correlation coefficient r s was 0.983. This method was used to detect neutralizing antibody titers in mouse sera against HRSV subtype A strain long and subtype B strain 9320, and the fusion proteins combined with AlOH and CpG adjuvant induced the highest neutralizing antibody titers in mice. Conclusion:The HRSV neutralizing antibody assay established in this study is rapid, reproducible, high-throughput, and can be used to detect neutralizing antibodies to HRSV subtypes A and B.

Aim To investigate the effect of alpinetin on the biological behavior of lung cancer cells,and fur-ther to analyze the relation between its anti-tumor mechanism and the regulation of miR-654-3p/high mobility group protein B1(HMGB1)axis.Methods The lung cancer A549 cells were divided into 0 μmol·L-1 group,alpinetin(50,100,200 μmol·L-1)group,miR-NC group,miR-654-3p mimic group,si-NC group,si-HMGB1 Group,alpinetin+miR-654-3p in-hibitor group,alpinetin+pcDNA-HMGB1 group.Cell counting kit-8(CCK-8)method and flow cytometry were performed to detect the inhibition rate and apopto-sis rate respectively.Plate cloning assay and Transwell chamber method were applied to detect the numbers of clone formation and migration.Quantitative reverse transcription polymerase chain reaction(RT-qPCR)was employed to analyze the expression of miR-654-3p and HMGB1 mRNA.Dual luciferase reporter assay was used to confirm the targeting effect of miR-654-3p on HMGB1.Western blotting was performed to detect pro-tein expression.Results Compared with the 0 μmol·L-1 group,the inhibition rate,apoptosis rate and miR-654-3p expression of A549 cells in the alpinetin group(50,100,200 μmol·L-1)increased,while clone formation numbers,migration numbers and HMGB1 expression were reduced(P＜0.01).Com-pared with the miR-NC group,the inhibition rate and apoptosis rate of A549 cells in the miR-654-3p mimic group increased,while clone formation numbers,migra-tion number and HMGB1 expression decreased(P＜0.01).Compared with the si-NC group,the inhi-bition rate and apoptosis rate of A549 cells in the si-HMGB1 group increased,while clone formation num-bers and migration were reduced(P＜0.01).MiR-654-3 p directly and specifically bound to HMGB1.Compared with the alpinism group,the inhibition rate and apoptosis rate of A549 cell and BCL2-associated x protein(Bax)and E-cadherin protein expression in al-pinetin+miR-654-3p inhibitor group,alpinetin+pcD-NA-HMGB1 group were reduced,while clone formation numbers,migration and B-cell lymphoma-2(Bcl-2)and N-cadherin protein expresison increased(P＜0.01).Conclusion Alpinetin exerts anti-prolif-eration,anti-migration and pro-apoptosis effects in lung cancer,and its mechanism is related to the up-regula-tion of the miR-654-3p/HMGB1 axis.

Objective: To explore the epidemiological characteristics and treatment outcomes of patients with hydrofluoric acid burns in hands. Methods: A retrospective observational study was conducted. The medical records of 229 patients with hydrofluoric acid burns in hands who were admitted to Zhejiang Quhua Hospital from January 2008 to December 2020 and met the inclusion criteria were collected. The following statistical data of patients were collected, including gender, age, type of affiliated enterprise, hydrofluoric acid mass fraction, injury site, total burn area, prehospital time, length of hospital stay, length of wound healing, whether hypocalcemia and hypomagnesemia occurred or not on admission, whether surgery intervention was performed or not, and whether scar sequelae occurred or not. Single factor and multivariate logistic regression analysis were used to screen out the risk factors impacting surgery intervention and scar sequelae of all the patients and patients whose hydrofluoric acid mass fraction was known. Single factor and multivariate linear regression analysis were used to screen out the risk factors impacting the length of wound healing of all the patients and patients whose hydrofluoric acid mass fraction was known. Results: The 229 patients included 206 males and 23 females, with the majority aged 30 to 50 years (139 patients). The type of affiliated enterprise of majority patients was non-fluorine chemical enterprise. The hydrofluoric acid mass fraction was known in only 91 patients, mainly medium. The majority injury site was in the middle and end of finger. The total burn area was below or equal to 1% total body surface area. The prehospital time was 19 (9, 29) h. The length of hospital stay was 2 (1, 7) d. The length of wound healing was 12 (8, 18) d. The proportions of hypocalcemia and hypomagnesemia were 0.9% (2/229) and 1.3% (3/229) on admission, respectively. Thirty-six patients had surgeries and 83 patients had scar sequelae. In 229 patients, single factor logistic regression analysis showed that both type of affiliated enterprise and prehospital time were the factors impacting surgery intervention (with odds ratio values of 7.86 and 51.35, respectively, 95% confidence intervals of 1.83-33.76 and 11.89-221.78, respectively, P<0.01) and scar sequelae of patients (with odds ratio values of 3.62 and 27.40, respectively, 95% confidence intervals of 1.76-7.43 and 13.25-56.68, respectively, P<0.01); multivariate logistic regression analysis showed that prehospital time was the independent risks factor impacting surgery intervention and scar sequelae of patients (with odds ratio values of 43.00 and 24.55, respectively, 95% confidence intervals of 9.89-187.03 and 11.78-51.16, respectively, P<0.01); single factor linear regression analysis showed that both type of affiliated enterprise and prehospital time were the factors impacting the length of wound healing of patients (with β values of 6.16 and 12.83, respectively, 95% confidence intervals of 3.38-8.93 and 10.72-14.93, respectively, P<0.01); multivariate linear regression analysis showed that both type of affiliated enterprise and prehospital time were the independent risk factors impacting the length of wound healing of patients (with β values of 2.81 and 12.16, respectively, 95% confidence intervals of 0.50-5.13 and 10.00-14.31, respectively, P<0.05 or P<0.01). In 91 patients whose hydrofluoric acid mass fraction was known, single factor logistic regression analysis showed that type of affiliated enterprise, hydrofluoric acid mass fraction (low and high), and prehospital time were all the factors impacting surgery intervention of patients (with odds ratio values of 9.10, 11.25, 10.69, and 0.04, respectively, 95% confidence intervals of 1.15-72.25, 1.39-90.93, 1.32-86.59, and 0.01-0.19, respectively, P<0.05 or P<0.01), type of affiliated enterprise, hydrofluoric acid mass fraction, and prehospital time were all the factors impacting scar sequelae of patients (with odds ratio values of 0.32, 0.21, and 36.80, respectively, 95% confidence intervals of 0.11-0.92, 0.06-0.73, and 11.03-122.79, respectively, P<0.05 or P<0.01); multivariate logistic regression analysis showed that both hydrofluoric acid mass fraction and prehospital time were the independent risk factors impacting surgery intervention of patients (with odds ratio values of 11.51 and 0.04, respectively, 95% confidence intervals of 1.22-108.26 and 0.01-0.25, respectively, P<0.05 or P<0.01), prehospital time was the independent risk factor impacting scar sequelae of patients (odds ratio=37.71, with 95% confidence interval of 9.97-142.69, P<0.01); single factor linear regression analysis showed that type of affiliated enterprise, hydrofluoric acid mass fraction (low and high), and prehospital time were all the factors impacting the length of wound healing of patients (with β values of 7.12, -5.63, -9.74, and 13.50, respectively, 95% confidence intervals of 2.43-11.81, -10.59--0.68, -14.78--4.70, and 10.14-16.86, respectively, P<0.05 or P<0.01); multivariate linear regression analysis showed that both hydrofluoric acid mass fraction and prehospital time were the independent risk factors impacting the length of wound healing of patients (with β values of -5.84 and 0.09, respectively, 95% confidence intervals of -10.59--1.08 and 0.05-0.12, respectively, P<0.05 or P<0.01). Conclusions: The majority of patients with hydrofluoric acid burns in hands are young and middle-aged males. Type of affiliated enterprise, hydrofluoric acid mass fraction and prehospital time are the risk factors that affect the treatment outcomes of patients with hydrofluoric acid burns in hands.
Adult ; Body Surface Area ; Burns ; Female ; Humans ; Hydrofluoric Acid/adverse effects* ; Length of Stay ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

Hydrofluoric acid is a highly dangerous and toxic inorganic acid, which is widely used in industrial fields and daily life. The risk of hydrofluoric acid burns is related to hydrofluoric acid mass fraction, duration of exposure to hydrofluoric acid, burn area, burn depth, and burn site, etc. Hydrofluoric acid has strong toxicity and tissue penetration ability. A small area of hydrofluoric acid burns can cause death in a short time. Therefore, improving the understanding of the mechanism of hydrofluoric acid burns and learning how to treat hydrofluoric acid burns in different sites can further improve the cure rate of hydrofluoric acid burns.
Burns, Chemical/therapy* ; Calcium Gluconate ; Humans ; Hydrofluoric Acid/adverse effects*