Objective To explore the effect and mechanism of Kuntai Capsule (KC) on premature ovarian failure in rats.Methods Totally 40 SD female rats were randomly divided into four groups:control group,model group,Kuntai Capsule (KC) group and conjugated estrogens tablets (CET) group.The premature ovarian failure model in rats was made by ig administration of 75 mg/kg tripterygium.After the model was established,rats were continually treated with 0.6 and 0.625 g/kg of KC and CET respectively by ig administration for 36 d.HE staining was used to observe the morphology of ovary and count the number of follicle.The enzyme-linked immunosorbent assay (Elisa) method was used for the detection of follicle stimulating hormone (FSH),luteinizing hormone (LH),serum estradiol (E2),and anti-Mueller tube hormone (AMH) level.The real-time fluorescence quantitative PCR (qRT-PCR) method was used to detect the mRNA expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF).Results Compared with control group,the number of primordial follicles,preantral follicles and antral follicles were significantly reduced,the number of atretic follicles was increased (P ＜ 0.05),E2 and AMH significantly decreased (P ＜ 0.05),FSH and LH levels increased significantly (P ＜ 0.05),and the levels of VEGF and bFGF mRNA in model group were significantly decreased (P ＜ 0.05).Compared with model group,the number of primordial follicles,preantral follicles,and antral follicles were significantly increased,the number of atretic follicles was significantly reduced (P ＜ 0.05),E2 and AMH significantly increased (P ＜ 0.05),FSH and LH levels significantly decreased (P ＜ 0.05),and the levels of VEGF and bFGF mRNA in KC group were significantly increased (P ＜ 0.05).Conclusion By regulating the level of hormone and up-regulating the expression of VEGF and bFGF,KC can repair the damaged ovarian tissue and promote the growth and development of the follicle,so as to inhibit the premature exhaustion of mRNA.

Objective:To identify the key genes related to rheumatoid arthritis (RA) by to the weighted gene co-expression network analysis (WGCNA) and experimental verification to find key genes related to RA.Methods:The microarray data of RA were downloaded from the Gene Expression Omnibus (GEO) database. Gene network was constructed, and the genes were classified into different modules using WGCNA. HUB genes in modules related to RA clinical symptoms were analyzed by gene ontology. Subsequently, different data sets of GEO were used to verify the expression profile and diagnostic capacity of the HUB gene [receiver operating characteristic curve (ROC)]. In addition, the expression of HUB gene in RA was verified by real time polymerase chain reaction (RT-PCR) and Western blot, and the relationship between key genes and disease activity score 28 joints (DAS28) was analyzed. Paired-sample t-test and Pearson's correlation analysis was used for statistical analysis. Results:A total of 5 413 differentially expressed genes were filtered. Weighted gene coexpression network was constructed and genes were classified into 23 modules. Among them, the black module is closely related to the clinical symptoms of RA, which contained 346 genes. Enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) signal pathway analysis showed that it was to be enriched in the positive regulation of interleukin 6, interleukin 1 beta secretion, osteoclast differentiation, NOD-like receptor signaling pathway, T helper cell 17 (Th17) cell differentiation and many other pathways closely related to RA. Motile sperm domain-containing protein 2 (MOSPD2) was significantly correlated with clinical symptoms. It was highly expressed in blood monocytes and bone marrow monocytes ( t=2.238, P=0.032; t=3.153, P=0.006), and positively correlated with blood expression in RA joint synovial fluid ( r=0.683, P=0.03). ROC curve analysis determined that MOSPD2 could distinguish RA from the control group (the area under the curve was 0.855 and 0.726) respectively. RT-PCR and Western blotting results showed that MOSPD2 was up-regulated in RA patients ( t=-3.96, P=0.02). MOSPD2 expression levels in blood were positively correlated with DAS28 in RA patients ( r=0.884 6, P=0.046 2). Conclusion:MOSDP2 is closely related to the clinical symptoms of RA patients, and may be one of the targets for the diagnosis and treatment of RA.

AIM: To investigate the effects of metformin on airway inflammation, remodeling and neovascularization in a mouse model of chronic asthma and its possible mechanisms.METHODS: BALB/c mice were randomly divided into saline group, ovalbumin (OVA) group and OVA+metformin group, with 8 in each.At the end of OVA exposure, blood and bronchoalveolar lavage fluid (BALF) were collected for the measurement of OVA specific IgE and leukocyte counts.Lung tissue sections were stained with hematoxylin-eosin, periodic acid-Schiff and Masson's trichrome to detect inflammatory cell infiltration, goblet cell hyperplasia, and collagen deposition around the airway, respectively.Immunohistochemistry was used to evaluate the number and percentage area of new blood vessels (CD31+), and the protein level of phosphorylated AMP-activated protein kinase (p-AMPK) in the airway.RESULTS: Compared with saline group, the eosinophil percentage and OVA specific IgE in serum in OVA group were all increased obviously (P<0.01).Metformin inhibited the above increases (P<0.05).Compared with control group, a marked increase in inflammation infiltration, PAS+ cells and collage deposition in the airway mucosa in OVA group were observed.Metformin partially relieved the above changes.CD31+ vessels in the wall of bronchi showed the abundance of blood vessels observed in OVA group compared with control group, which was suppressed by the treatment with metformin (P<0.05).The protein level of p-AMPK was reduced in the lung tissue challenged with OVA as compared with control group (P<0.05), while metformin increased the protein level of p-AMPK (P<0.01).CONCLUSION: The protein level of p-AMPK in the airway in OVA group is attenuated.Metformin effectively inhibits airway inflammation, remodeling and neovascularization possibly via activating AMPK signaling pathway.

Objective To analyze the relationship between thyroid function and grades of diabetic nephropa-thy (DN).Methods One hundred and twenty patients with definite DN were classified into the four groups,and forty patients with non-DN diabetic was NDN group.Their serum albumin,blood hemoglobin,serum potassium,functional parameters of thyroid and dependability to DN were analyzed.Results The plasma albumin ofⅢ,Ⅳ and Ⅴ group were (41.3 ±3.6)g/L,(30.5 ±4.8)g/L and (28.3 ±5.9)g/L,which were lower than those of NDN group (F=11.36,P<0.05).24h urine protein ofⅢ,Ⅳ and Ⅴ were (0.48 ±0.29)g,(1.86 ±0.54)g and (1.69 ±0.67)g, which were higher than those of group NDN (F=7.12,P<0.05).Triiodothyronine(T3)ofⅢ,ⅣandⅤgroup were (0.98 ±0.38)nmol/L,(0.75 ±0.41)nmol/L and (0.60 ±0.28)nmol/L,which were lower than those of NDN group (F=7.64,P<0.05).Tetraiodothyronine(T4)of III,IV and V group were (6.06 ±1.76)nmol/L,(5.31 ±1.98)nmol/L and (4.65 ±1.87)nmol/L,which were lower than those of NDN group (F=6.83,P<0.05 ).Free Triiodothyronine (FT3)ofⅢ,Ⅳ and Ⅴ group were (4.37 ±2.12)pmol/L,(3.33 ±2.30)pmol/L and (2.91 ±1.82)pmol/L, which were lower than those of NDN group (F=7.14,P<0.05 ).Correlation analysis indicated that FT3 was an independent predictor for the severity of diabetic nephropathy.Conclusion In the patients with DN Ⅳ and Ⅴ,DN has attained a very severe status and the treatment of DN should be intensive.

Objective : To examine the association between acute exposure to traffic-related air pollutants (TRAP) NOX and NO2 and outpatient visits of pediatric respiratory diseases. Methods : Data regarding outpatient visits to Department of Respiratory Diseases of Beijing Children's Hospital from 2015 to 2020 were collected, and the concentrations of nitrogen oxides (NOX), nitrogen dioxide (NO2) and other TRAP were collected from the surveillance sites assigned by the Peking University Health Science Center. A time-stratified case-crossover design was employed, and a conditional logistic regression model was created to examine the association between NOX and NO2 acute exposure and outpatient visits of pediatric respiratory diseases. Results : The daily mean outpatient visits of pediatric respiratory diseases were 571 (interquartile range, 554) person-times among children at ages of 0 to 14 years in Beijing Children's Hospital from 2015 to 2020, and the daily mean outpatient visits for upper respiratory tract infections (URI), bronchitis, and pneumonia were 265 (interquartile range, 282), 143 (interquartile range, 178) and 128 (interquartile range, 120) person-times, respectively. The daily mean concentrations of atmospheric NOX and NO2 were 67.8 (interquartile range, 50.7) and 49.3 (interquartile range, 30.7) μg/m3, respectively. Conditional logistic regression analysis showed the largest lagged effect of NOX and NO2 on pediatric respiratory diseases at cumulative lags of 0 to 7 days. An increase in NOX concentrations by an interquartile range resulted in the excess risks of URI, bronchitis and pneumonia by 6.87% (95%CI: 6.37%-7.38%), 7.25% (95%CI: 6.51%-7.99%), and 5.51% (95%CI: 4.69%-6.33%), and an increase in NO2 concentrations by an interquartile range resulted in excess risks of URI, bronchitis and pneumonia by 5.71% (95%CI: 5.12%-6.31%), 5.32% (95%CI: 4.51%-6.14%), and 4.83% (95%CI: 3.91%-5.75%), respectively. NOX and NO2 presented a more remarkable effect on outpatient visits of pediatric respiratory diseases among children at ages of over 5 years. Conclusion NOx and NO2 acute exposure may increase the outpatient visits of pediatric respiratory diseases.

BACKGROUND:Embryonic stem cel s have the capacity of multi-differentiation potential, and have been utilized for the therapy of acute liver injury. However, the migration and proliferation of embryonic stem cel s after transplantation remains not wel characterized.
OBJECTIVE:To track the transplanted embryonic stem cel s in repairing acute liver injury by bioluminescence imaging technology.
METHODS:Murine embryonic stem cel s (D3) were transducted with a construct composed of firefly luciferase, monomeric red fluorescence protein and herpes simplex virus truncated thymidine kinase triple fusion reporter genes by lentivirus system. Stable D3 embryonic stem cel s integrating three report genes were screened. The undifferentiated embryonic stem cel s or differentiated embryonic stem cel s from the 6-day-old embryoid body were transplanted into acute liver injury model of SV129 mouse through spleen, and the transplanted cel s were monitored by bioluminescence imaging technology.
RESULTS AND CONCLUSION:Reverse transcription PCR results showed that the expression level of Oct-4 and Nanog was not affected in embryonic stem cel s transducted with triple fusion reporter gene compared with wild-type embryonic stem cel s. The migration process of transplanted cel s was visualized by bioluminescence imaging technology. Teratomas were found in both triple fusion-embryonic stem cel s treatment group and triple fusion-embryoid body cel s treatment group at liver, and the teratoma formation could be suppressed by ganciclovir administration because ganciclovir can react with herpes simplex virus truncated thymidine kinase and trigger cel necrosis process. Histological analysis showed that teratomas comprised tissues from al three germ layers. These results demonstrate that triple gene fusion does not affect differentiation potential of embryonic stem cel s and it is risky to utilize embryonic stem cel s for cel therapy, because it affects repair of liver injury. The therapy strategy requires further improvement and real-time visualizing of embryonic stem cel s in vivo is absolutely necessary.

Glucocorticoids/urine* ; Humans ; Hydrocortisone ; Marathon Running ; Saliva/chemistry* ; alpha-Amylases/analysis*

Objective: To investigate parents acceptance to the COVID-19 vaccine booster shots for their children aged 3-11 years in Nanjing based on the theory of planned behavior, and to provide a scientific theoretical basis for carrying out COVID-19 booster vaccination among children of this age group. Methods: A total of 1 286 parents of children aged 3-11 years in Nanjing were selected by multistage stratified cluster sampling. A questionnaire survey based on the theory of planned behavior was used to investigate their willingness to vaccinate their children with the COVID-19 booster dose, and structural equation model was used to conduct data analysis. Results: About 90.2% of parents were willing to give their children a booster dose of the COVID-19 vaccine. The model constructed based on the theory of planned behavior could explain 42.2% of the variance of vaccination acceptance. Attitudes and perceived behavioral control had a direct positive effect on parents acceptance to booster dose of the COVID-19 vaccine (path coefficients were 0.47 and 0.18, P <0.01); The direct effect of subjective norms on vaccination acceptance was not statistically significant; Perceived behavioral control and subjective norms could have indirect positive effects on vaccination acceptance through attitudes (path coefficients were 0.27 and 0.13, P <0.01). Conclusion Parents in Nanjing have a higher acceptance to vaccinate their children aged 3-11 years with the COVID-19 vaccine booster dose. The theory of planned behavior shows a good explanatory ability on parents acceptance to vaccinate their children, and attitude plays an important role in the formation of vaccination acceptance.

BACKGROUND:Currently,different methods of model establishment have been derived from different injury modes of spinal cord injury.Traditional physical injury modeling methods have their own advantages and disadvantages,and there is a lack of more effective and stable animal models of spinal cord injury. OBJECTIVE:To establish a reproducible,controllable,trauma-free,low-mortality,more stable,widely applicable,and short-term postoperative care rat model of spinal cord injury. METHODS:Forty Sprague-Dawley rats with similar body mass and ages were randomly divided into a control group and an improved group,with 20 rats in each group.Animal models of spinal cord injury in the control group were constructed using a clip model method,while the improved group used a modified ligation method based on the compression method to make the spinal cord injury models using suture ligation based on fenestration.Postoperative comparisons were made between the two groups,assessing urination behavior,hematuria,pyuria(infection rate),mortality,scoliosis rate and Basso-Beattie-Bresnahan locomotor rating scale scores at 1,3,5,and 7 days after modeling. RESULTS AND CONCLUSION:Compared with the conventional modeling method,the modified ligation method based on the compression method resulted in faster recovery of urination behavior,lower hematuria rate,lower infection rate,lower mortality rate,lower scoliosis rate,and more concentrated and stable Basso-Beattie-Bresnahan scores(all below 2 points within 1 week).This proves that the modified ligation method based on compression is more suitable for the establishment of spinal cord injury models in rats.

Objective To identify the potential long non-coding RNA (lncRNA) expressed in rheumatoid arthritis (RA) synovium key to RA onset and investigate its association with immune cell infiltration. Methods RA synovium data were downloaded from the GEO database and normalized. The lncRNAs key to RA onset were identified using multiple machine learning methods. Infiltration of 22 immune cell populations in RA synovium was measured by cell-type identification by estimating relative subsets of RNA transcripts (CIBER-SORT). The relationship between the key lncRNA and infiltrating immune cells was analyzed. Finally, real-time quantitative PCR was applied to validate the expression of the key lncRNA in RA synovial cells. Results lncRNA human leukocyte antigen complex P5(HCP5) was identified as the key lncRNA associated with RA onset. Infiltration analysis revealed increased abundance of CD8⁺ T cells, γδ T cells, and M1 macrophages while decreased abundance of M2 macrophages in RA synovial tissue. Correlation analysis demonstrated that the lncRNA HCP5 expression was positively associated with the infiltration abundance of CD8⁺ T cells, γδ T cells, and M1 macrophages in RA synovial tissue. Furthermore,the expression of lncRNA HCP5 in RA synovial cells was up-regulated. Conclusion lncRNA HCP5 expression is up-regulated in RA synovial tissue and potentially associated with immune cells infiltration.
Humans ; Arthritis, Rheumatoid ; CD8-Positive T-Lymphocytes ; HLA Antigens/metabolism* ; RNA, Long Noncoding/metabolism* ; Synovial Membrane/metabolism*