Bio-mineralization has emerged as a promising strategy to enhance vaccine immunogenicity. This study optimized the calcium phosphate (CaP) mineralization process of foot-and-mouth disease virus-like particles (FMD VLPs) to achieve high mineralization efficiency and scalability. Key parameters, including concentrations of Ca²⁺, HPO₄^2-, NaCl, and VLPs, as well as stirring speed, were systematically optimized. Stability of the scaled-up reaction system and immunogenicity of the mineralized vaccine were evaluated. Optimal conditions [25.50 mmol/L Ca(NO₃)₂, 15 mmol/L Na₂HPO₄, 300 mmol/L NaCl, 0.75 mg/mL VLPs, and 1 500 r/min] yielded CaP-mineralized VLPs (VLPs-CaP) with high mineralization efficiency, uniform morphology, and a favorable particle size. Scaling up the reaction by 25 folds maintained consistent mineralization efficiency and particle characteristics. Immunization in mice demonstrated that VLPs-CaP induced higher titers of specific antibodies and neutralizing antibodies than unmineralized VLPs (P < 0.05). Higher IgG2a/IgG1 ratio and enhanced IFN-γ secretion (P < 0.05) further indicated robust cellular immune responses. We establish a stable and scalable protocol for VLPs-CaP, providing a theoretical and technical foundation for developing high-efficacy VLPs-CaP vaccines.
Vaccines, Virus-Like Particle/immunology* ; Immunogenicity, Vaccine ; Calcium Phosphates/chemistry* ; Foot-and-Mouth Disease Virus ; Biomineralization ; Particle Size ; Animals ; Mice ; Antibodies, Neutralizing/blood* ; Antibodies, Viral/blood* ; Immunity, Cellular

Animals ; Macaca mulatta ; Optic Disk ; Glaucoma ; Craniocerebral Trauma ; Intraocular Pressure ; Low Tension Glaucoma

Objective : To explore the effects of microRNA-155 (miR-155) on apoptosis of chronic myeloid leukemia ( CML) cells，and the influence of miRNA-155 regulating the expression of heat shock proteins ( HSP) 27， HSP60，HSP70． Methods : Reverse transcription-quantitative real-time PCR ( RT-qPCR) was used to detect the expression levels of miR-155 in CML-resistant imatinib (IM) cell line K562-G and CML cell line K562 ．K562-G cells were infected with the lentivirus carrying miR-155 or the negative control lentivirus ，and they were named miR-155 group and control group．The effect of miR-155 on the proliferation of drug-resistant cells was detected by cell counting kit-8 ( CCK-8) method. RT-qPCR and Western blot were used to detect the effect of miR-155 on the expression of heat shock proteins HSP27，HSP60，HSP70．Flow cytometry was used to detect the percentage of cell apoptosis in miR-155 group and control group. Results : ompared with K562 cells，miR-155 showed low expres- sion in K562-G cells (P ＜0. 05) ．The proliferation of miR-155 group cells decreased significantly from the 36th hour (P＜0. 05) ．Compared with the control group，in the miR-155 group，HSP60 and HSP70 increased (P < 0. 05) ，while HSP27 decreased (P＜0. 01) ．The apoptosis rate of miR-155 group was higher than that of control group (P＜0. 05) ． Conclusion miR-155 promotes the apoptosis of chronic myeloid leukemia cells，increases the expression of HSP60 and HSP70，and decreases the expression of HSP27 .

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.

Objective To conduct nutrition risk screening on newborns with modified screening tool for the assessment of malnutrition in pediatrics (STAMP), and to analyze the correlation between nutritional risk of newborns and diseases. Methods A total of 2 300 newborns treated in Neonatal Center (NICU, Department of Neonatology and Department of Neonatal Surgery) in Children's Hospital of Shanxi from February to December 2015 were screened by modified scale STAMP about their nutritional risks, with the data collected and analyzed. Results Top five high risks concerning disease and nutrition among newborns were: hyaline membrane, volvulus, hematochezia of unknown origin, premature infants, and abdominal distension of unknown origin. Differences of malnutrition conditions and nutrition intake among newborns from different departments were statistically significant (χ2=31.74, 177.00; P<0.005). Majority of newborns in NICU and Department of Neonatal Surgery were without any nutrition intake at all, while majority of newborns in Department of Internal Medicine were with only half or less nutrition intake. Differences of high malnutrition risks and insufficient nutrition intake among premature and term infants from different departments were statistically significant (χ2=72.70, 53.95; P<0.001). Differences of weight growth conditions among newborns from different departments were statistically significant (χ2=13.57, P=0.002). Conclusions By using modified STAMP, it can be identified that factors causing high risks of neonatal nutrition include diseases, gestational age, nutritional intake, growth and development. Individualized nutritional support program should be implemented according to results of the screening, so that status quo of extrauterine growth restriction can be improved.

Phenylalanine hydroxylase (PAH) is a member of aromatic amino acid hydroxylase (AAAHs) family, and catalyze phenylalanine (Phe) into tyrosine (Tyr). Using immunological and RT-PCR methods to prove the existence of phenylalanine hydroxylase (PAH) gene in the brain of Neanthes japonica in protein and nucleic acid level. Using Western blotting to detect the pah immunogenicity of Neanthes japonica. Making paraffin sections and using immunohistochemical technique to identify the presence and distribution of the phenylalanine hydroxylase gene in the brain of Neanthes japonica. Clone pah gene from the brain of Neanthes japonica by RT-PCR, constructing plasmid and transferring into E. coli to amplification, picking a single homogeneous colony, double digesting then making sequence and comparing homology. Western blotting results showed that the expression of the protein is present in Neanthes japonica brain, immunohistochemistry technique results showed that phenylalanine hydroxylase mainly expressed in abdominal of forebrain, dorsal and sides of midbrain. RT-PCR technique results showed that the phenylalanine hydroxylase exist in the brain of Neanthes japonica and has a high homology with others animals. PAH is present in the lower organisms Neanthes japonica, in protein and nucleic acid level. Which provide the foundation for further study the evolution of aromatic amino acid hydroxylase genes in invertebrate.
Animals ; Blotting, Western ; Brain ; enzymology ; Escherichia coli ; metabolism ; Phenylalanine Hydroxylase ; genetics ; metabolism ; Polychaeta ; enzymology ; genetics

Objective:To screen the new candidate molecules interacting with protein kinase Wee1B by yeast two hybrid system, and to analyze their interaction with Wee1B in the early stage of mouse fertilized eggs by bioinformatics.Methods:The plasmid pcDNA3.1/V5-His-TOPO-Wee1B wild type encoding mouse Wee1B gene was used as template to construct bait plasmid pGBKT7 Wee1B and the bait plasmid pGBKT7-Wee1B was transformed into yeast competent cells at SD/Trp (SDO),SD/Trp/X-α-Gal (SDO/X)and SD/Trp/X α Gal/AbA plates (SDO/X/A)plates to detect the toxicity and self-activation ability of yeast and its expression in yeast using Western blotting method.The yeast cells containing pGBKT7-Wee1B were fused with human ovary cDNA library, the yeast plasmid transformation of Escherichia coli positive clones were sequenced after identified by yeast transformation.BLAST analysis was carried out in GenBank,and its effect on the development of mouse fertilized eggs was deduced according to the gene annotation.Results:The double enzyme digestion analysis and sequencing analysis results showed that the pGBKT7-Wee1B bait plasmid was successfully constructed.The plasmid was transformed into the yeast,and there were no clones in the SDO/X/A plates.The pGBKT7-Wee1B and pGBKT7 empty vectors were transformed into the yeast,the bacteria were inoculated on the SDO plates,and the clones were uniformly grown on the two SDO plates.The positive clones were picked and expanded in culture,the protein was extracted and Western blotting showed that pGBKT7 Wee1B was expressed in the yeast.The bait plasmids were fused with human ovary cDNA library and the positive clones inserted into the fragment were identified by PCR. Nine proteins which interacted with Wee1B protein kinase were screened out by sequencing and blast analysis,and the proteins which could be closely related to the development of mouse oocytes and the development of fertilized eggs were analyzed by bioinformatics analysis.Conclusion:Using the yeast two hybrid system from human ovary cDNA library,nine interacting proteins with Wee1B protein kinase are screened and these screened proteins may regulate mouse oocyte maturation and early embryo development through interacting with Wee1B.

BACKGROUND: It is a hotspot that calcium phosphate and calcium sulphate as the main ingredients are combined with one or more other materials to improve or increase the performance of bone tissue engineering scaffolds. OBJECTIVE: To introduce the research advance of these two kinds of scaffolds in bone tissue engineering. METHODS: The articles related to the bone tissue engineering published during January 2000 to June 2015 were retrieved from CNKI and PubMed databases by computer. The key words were “bone tissue engineering, scaffold, calcium phosphate, calcium sulphate, vascularization” in Chinese and English, respectively. ESULTS AND CONCLUSION: Calcium phosphate and calcium sulfate are characterized as having good biocompatibility, biodegradability, osteoconductivity and complete bone substitutability. However, single use of calcium phosphate or calcium sulfate scaffold has certain disadvantages, both of which are difficult to ful y meet the requirements of the bone defect repair. Improvement can be acquired in the mechanical strength, injectability and biodegradability, as wel as drug-loading and pro-angiogenesis of the scaffold in combination with other materials. In the basal and clinical research, we should explore and develop ideal scaffolds in on the basis of therapeutic aim. However, most of the scaffold studies are stil at the extracorporeal and animal experiment stage, and the comparative studies on composite scaffolds and optimal proportion of those composite scaffolds stil need to be further investigated.

BACKGROUND:There are so many studies about ovariectomized rats at present, but the research on the change rules of bone mass, bone turnover markers, estrogen levels and their correlation in different periods of ovariectomized rats is rarely reported. OBJECTIVE:To analyze the change rules of bone mass, bone turnover markers, estrogen level and their correlation in different periods of ovariectomized rats. METHODS: Thirty-four 3-month-old female Sprague Dawley rats were randomly divided into three groups: baseline group, ovariectomized group and sham operated group. At the beginning of the experiment, the rats in the baseline group were sacrificed, then rats in the ovariectomized group and sham operated group were executed at 4, 8, 12 weeks postoperative respectively. The bone mineral density, bone mass content, area of different zones of the L1-3 lumbar vertebrae and femurs were detected by dual-energy X-ray absorption method, and meanwhile the serum levels of type I procolagen amino-terminal pro-peptide, I colagen carboxy-terminal peptide and estrogen were determined by ELISA. At last, we analyzed the correlation between body mass, bone mineral densityin vitro, type I procolagen amino-terminal pro-peptide, I colagen carboxy-terminal peptide and estrogen levels and the age of ovariectomized rats. RESULTS AND CONCLUSION: (1) The bone mineral density and bone mass content of the lumbar vertebral and femurs in the ovariectomized group were significantly lower than those in the sham operated group and baseline group at the 4th week after operation (P < 0.05). The bone mineral density and bone mass content in the ovariectomized group were ameliorated obviously at the 8th and 12th weeks compared with those at the 4th week after operation (P < 0.05). The bone mass loss was highest in the L1 and intertrochanteric regions. (2) Serum levels of type I procolagen amino-terminal pro-peptide and I colagen carboxy-terminal peptide in the ovariectomized group were significantly higher than those in the baseline group and sham operated group at the 4th week after operation, but there was no difference at the 8th and 12th weeks. (3) The serum estrogen level in the ovariectomized group was prominently lower than that in the sham operated group and baseline group at the 8th and 12th weeks after operation (P < 0.01 at the 8th week,P < 0.05 at the 12th week). (4) The age was positively correlated with body mass and bone mineral density of the lumbar vertebrae and femursin vitro, while the serum levels of type I procolagen amino-terminal pro-peptide and I colagen carboxy-terminal peptide were negatively correlated with the bone mineral density of the lumbar vertebrae and femurs in vitro (P < 0.01). These results suggested that the bone mass of the lumbar vertebrae and femurs in ovariectomized rats was decreased rapidly firstly, and then rose slowly with time; the bone mass in the L1 and intertrochanteric regions lost seriously; the bone turnover markers showed a significant increase at the beginning of ovariectomy and reduced gradualy to normal condition, while the estrogen level was increased at the first month after ovariectomy and then decreased rapidly. In addition, the body mass, bone turnover markers and estrogen level were associated with the change of bone mass.

Objective To compare the effects of gastric gavage and intramuscular injection of prednisone on the bone mineral density, skeletal biomechanical properties and bone metabolism in rats.Methods A total of 45 SPF rats were randomly divided into three groups:normal group, intragastric administration group, and intramuscular injection group.The normal group, as a control group, was administrated with normal saline 2 mL per day, both the intragastric administration group and i.m.injec-tion group received prednisone 0.5 mg/(kg.d) for 12 weeks.All rats were examined for bone mineral density (BMD) and the level of serum β-CTX and PINP.The femoral cortical biomechanical properties ( elastic load, maximal load, rupturing load) were measured by three point bending test.Results After 12 weeks, compared with the normal group, BMD and elastic load, maximal load, and rupturing load of the femur were significantly decreased.Compared with the intragastric gavage group, BMD was significantly decreased, while the elastic load, maximal load, and rupturing load of the femur were not significantly changed in the i.m.injection group (P＜0.05 for all).Compared with the normal group, the level of serum β-CTX was significantly raised (P＜0.05) and the level of serum PINP was significantly decreased (P＜0.05).Compared with the intragastric gavage group, the level of serumβ-CTX was also significantly raised (P＜0.05), the level of serum PINP was significantly decreased (P＜0.05), the bone trabecula and hemopoietic tissue were obviously decreased, while the adipose tissue increased obviously. Conclusions Both intragastric gavage and intramuscular injection of prednisone affect the level of BMD, skeletal biomechanical properties and bone metabolism.However, i.m.injection of prednisone decreases the BMD and bone strength more significantly, leading to a higher bone turnover with increased bone resorption, and leads to osteoporosis earlier.Our results may suggest that oral administration of prednisone is more safe in clinical treatment.