BACKGROUND:Numerous studies have shown that mesenchymal stem cel s (MSCs) can effectively attenuate the fibrosis of damaged heart, lung and kidney by secreting various bioactive factors. OBJECTIVE:To evaluate the anti-fibrotic therapeutic potential of bone marrow MSCs conditioned media in vitro. METHODS:Normal fibroblasts and hypertrophic scar fibroblasts were treated with bone marrow MSCs conditioned media, then transforming growth factor-βand col agen production were analyzed by ELISA, and mRNA expression level of Smad7 and hydroxyproline content were detected by RT-PCR and colorimetry, respectively. RESULTS AND CONCLUSION:Bone marrow MSCs conditioned media significantly inhibited the production of both transforming growth factor-βand col agen in hypertrophic scar fibroblasts (P<0. 01), and up-regulated the mRNA expression level of Smad7 (P<0. 01), a major inhibitory regulator in the SMAD family. However, the normal fibroblasts were scarcely influenced by bone marrow MSCs conditioned media. These findings indicate that bone marrow MSCs conditioned media is considered a promising candidate for the treatment of hypertrophic scars, which may provide new theoretical supports to reduce cutaneous scarring.

Hematopoietic stem cell transplantation in children has the characteristics of less stem cell for transplantation and higher acceptability of donors. The article reviews the classification, clinical application, general planning and clinical advantages of hematopoietic stem cell transplantation in children, to provide a new treatment for children's immune system diseases and genetic and metabolic diseases.

Objective: To develop a real-time quantitative PCR (qPCR) assay for detecting Sendai virus gene copy number. Methods: The recombinant plasmid was constructed and a reference used to establish the real-time qPCR method with the TaqMan probe and verified for reproducibility and sensitivity. Results: The linear range of the developed real-time qPCR assay was 1.024 × 10³ ~5 × 10¹⁰ copies/μl, with an R² value of more than 0. 99. The standard recovery of the tested samples was 84.56%~119.48%. Conclusions Compared with traditional hernagglutination assay for particle number detection, this method has higher sensitivity, better repeatability and high quantitative accuracy. It can be used for the detection of particle number of vaccine with sendai virus as the carrier, so as to detect the specific activity of vaccine products.

Pyroptosis is a kind of programmed cell death accompanied by inflammatory responses, which is mainly induced by the Caspase-1/4/5/11 activated mediated via inflammasomes or lipopolysaccharide.Pyroptosis causes the cleavage of various Gasdermin family members, including Gasdermin D, in which a large number of pro-inflammatory factors are released to cause inflammatory responses in various diseases.At present, pyroptosis has not been extensively analyzed in pediatric surgical diseases.This review summarizes the research progress of pyroptosis in pediatric surgical diseases like Hirschsprung′s disease, biliary atresia, neonatal necrotizing enterocolitis, and tumors.It is found that pyroptosis is involved in the occurrence and development of pediatric surgical diseases.The research on the specific mechanism of pyroptosis in pediatric surgical disease and involved signaling pathways contributes to the early diagnosis and treatment of some pediatric surgical diseases.

Objective:To construct a recombinant herpes simplex virus 2 (HSV-2) expressing enhanced green fluorescent protein (EGFP) using clustered, regularly interspaced, short palindromic repeat/CRISPR-associated nuclease 9 (CRISPR/Cas9) technology.Methods:Four strategies for inserting exogenous EGFP gene into HSV-2 genome using CRISPR/Cas9 technology were designed: (1) conventional homology-directed repair: circular two homology arm donor-mediated gene knock-in; (2) linearized single homology arm donor-mediated gene knock-in; (3) homology-independent targeted integration; (4) conventional homology-directed repair-mediated by cell lines stably expressing Cas9 and sgRNA.Results:The recombinant virus HSV-2-EGFP was successfully constructed based on the second, the third and the fourth strategies. The second strategy was the most efficient, followed by the third and the fourth strategies. The purified recombinant virus could stably express green fluorescent protein in seven passages and shared similar growth characteristics in Vero cells to the parental virus.Conclusions:Linearized single homology arm donor could increase the efficiency of gene knock-in, and cell lines stably expressing Cas9 and sgRNA could increase the efficiency of gene knock-in mediated by homology-directed repair.

Objective This study evaluated the effects of health education promoting therapeutic lifestyle changes in a population with dyslipidemia. Methods Patients with dyslipidemia were randomly assigned to one of four groups: the current group (CG) received conventional health guidance, the educational course (EC) group attended six lectures as part of an educational course, the phone call (PC) group received twice-monthly follow-up by telephone,and the comprehensive group(EC+PC)attended both the educational course and received follow-up telephone calls. Total cholesterol (TC), total triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein (HDL-C), and the knowledge, attitude, and behavior (KAP) score for blood lipids were compared within each group and among groups. Results A total of 214 patients were enrolled and completed the study: 62 patients in CG,49 patients in EC,56 patients in PC,and 47 patients in EC+PC.There were significant differences in the EC,PC,and EC+PC groups after the 24-week intervention. For example, pre- and post-intervention values for each group were as follows:EC group:(5.74±0.69)mmol/L and(5.14±0.87)mmol/L for TC,35.22±1.67 and 42.96±5.72 for KAP;PC group:(5.63±0.58)mmol/L and(5.22±1.07)mmol/L for TC, 34.54.±0.97and 39.41±5.03 for KAP;EC+PC group:(5.60±0.48)mmol/L and(4.00±0.79)mmol/L for TC,35.44±1.80 and 45.05±3.19 for KAP, respectively (P<0.05). The CG group showed no significant differences before and after treatment:(5.66±0.54)vs.(5.32±1.28)mmol/L for TC,34.37±0.65 vs.35.28±4.02 for KAP(P>0.05).In a comparison among the four groups,the EC and PC groups showed greater improvements than the CG group.Moreover, the EC+PC group showed statistically significant differences in the results compared with the other three groups (P< 0.05). Conclusion An educational course combined with telephone follow-up calls was more effective than a single intervention in improving blood lipids and enhancing the health awareness of patients with dyslipidemia.This combined health education model not only improves the effectiveness of treatment to some degree,but also plays a role in its supervision and management.Furthermore,it may also assist in the implementation of continuous nursing services in medical institutions.

Objective To investigate the influence of curcumin and its analogue H8 on glucose and lipid metabolism disorder in db/db mice. Methods The type 2 diabetes mouse model (db/db mice) was intragastrically administrated with curcumin and analogue H8 for 8 weeks.The blood biochemical indexes were measured.The expression of PEPCK and G6Pase mRNA was detected by real-time PCR in liver tissues.The expression of PEPCK and G6Pase protein was detected by Western blotting. Results Curcumin analogue H8 reduced blood glucose and lipids in db/db mice (P<0.01) and improved liver function related enzymes significantly.The levels of PEPCK and G6Pase mRNA in db/db mice were significantly decreased (P<0.01) and the expression levels of PEPCK and G6Pase protein were significantly decreased(P<0.01). Conclusion Curcumin analogue H8 improves the glucose and lipid metabolism disorder in db/db mice,and it is related to inhibiting the expression of PEPCK and G6Pase gene and protein.

Objective:To investigate the effects of genomic location of a foreign gene in Shanghai-191 strain measles virus (MV) vector on gene expression and virus replication.Methods:The nucleotide sequence encoding S1 protein of SARS-CoV-2 was inserted at different positions in MV antigenome (the upstream of the N gene, between P and M genes, between H and L genes), and co-transfected into 293T cells with helper plasmids coding T7 RNA polymerase and N, P, and L proteins, respectively. The transfected cells were lysed and the supernatants were used to infected Vero cells to harvest recombinant viruses. S1 proteins expressed by the recombinant viruses were identified by RT-PCR, indirect immunofluorescence assay, Western blot and ELISA. Growth kinetics of the recombinant viruses were analyzed.Results:Recombinant viruses were failed to be rescued when the S1 protein-coding sequence was cloned into the upstream of N gene. Two recombinant viruses, MV-M-S1 and MV-L-S1, were successfully rescued when cloning the S1 protein-coding sequence into the intergenic region between P and M genes, or H and L genes, and could express S1 protein. MV-M-S1 expressed more S1 protein than MV-L-S1, but the titer of MV-M-S1 was lower.Conclusions:Inserting a foreign gene at different positions in the MV genome might have different effects on gene expression and virus replication. This study provided reference for the subsequent construction of MV vector.

OBJECTIVES: This population-based, prospective cohort study investigated the association between glaucoma and mortality in older adults. METHODS: Participants aged 45 years or older at baseline (47.9% male) were enrolled in 2011 for the China Health and Retirement Longitudinal Study (CHARLS). All-cause mortality was observed during 7 years of follow-up. The baseline data were collected in the 2011 CHARLS, and participants were followed up for 7 years (until 2018). The risk of all-cause mortality was investigated using Cox proportional-hazards regression with age as the time scale, adjusting for significant risk factors and comorbid conditions. RESULTS: Among the 14,803 participants included, the risk of all-cause death was significantly higher among people with glaucoma than among those without glaucoma, after adjustment for other confounders (hazard ratio [HR], 1.46; 95% confidence interval [CI], 1.04 to 2.03). In a subgroup analysis based on the mean age of death, among those who were 75 years and older (n=1,231), the risk of all-cause death was significantly higher in patients with glaucoma than in those without glaucoma (HR, 1.89; 95% CI, 1.24 to 1.89). CONCLUSIONS Participants with glaucoma had a higher risk of all-cause mortality, especially those aged 75 years and above. Our findings revealed potential mechanisms underlying an association between glaucoma and all-cause mortality. They also highlighted the importance of glaucoma management to prevent premature death in middle-aged and older adults.

OBJECTIVES: This study evaluated the association between obesity and glaucoma in middle-aged and older people. A population-based retrospective cohort study was conducted using data from the China Health and Retirement Longitudinal Study. METHODS: Glaucoma was assessed via self-reports. Multivariate logistic regression analysis and a Cox proportional hazards model were used to assess the relationship between obesity and glaucoma risk. RESULTS: Older males living in urban areas who were single, smokers, and non-drinkers were found to have a significantly higher incidence of glaucoma (all p<0.05). Diabetes, hypertension, and kidney disease were also associated with higher glaucoma risk, while dyslipidemia was associated with lower risk (all p<0.05). After the model was adjusted for demographic, socioeconomic, and health-related variables, obesity was significantly associated with a 10.2% decrease in glaucoma risk according to the Cox proportional hazards model (hazard ratio, 0.90; 95% confidence interval [CI], 0.83 to 0.97) and an 11.8% risk reduction in the multivariate logistic regression analysis (odds ratio, 0.88; 95% CI, 0.80 to 0.97). A further subgroup analysis showed that obesity was associated with a reduced risk of glaucoma in people living in rural areas, in smokers, and in those with kidney disease (all p<0.05). Obesity also reduced glaucoma risk in people with diabetes, hypertension, or dyslipidemia more than in healthy controls (all p<0.05). CONCLUSIONS This cohort study suggests that obesity was associated with a reduced risk of glaucoma, especially in rural residents, smokers, and people with kidney disease. Obesity exerted a stronger protective effect in people with diabetes, hypertension, or dyslipidemia than in healthy people.