OBJECTIVETo investigate the development of xenografted primitive human germ cells by using fetal testicular tissues as donor tissues and an immunodeficient mouse as the recipient.

METHODSTesticular tissue fragments of a 26-week fetus were grafted under the back skin of a castrated immunodeficient mouse. Grafts were taken out after 135 days and processed for morphological and histological analyses.

RESULTSThe mass of grafts grew from about 1 mm in diameter and 5 mg in wet weight to about 3 mm and more than 20 mg 135 days after grafting. Histological observations showed a significant expansion of seminiferous tubules after grafting (80 +/- 25 microm in diameter) in comparison with seminiferous cords at the time of grafting (60 +/- 15 microm in diameter). The seminiferous cords developed into seminiferous tubules with the epithelial border and lumen. After 135 days of grafting, most of the dispersedly distributed primitive Sertoli cells and germ cells migrated to the basal part of seminiferous epithelium, located on the basement membrane and few of germ cells differentiated into spermatogonia.

CONCLUSIONHuman fetal testicular tissues could survive and continuously develop after being xenograft into castrated immunodeficient mice.

Animals ; Fetal Tissue Transplantation ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Spermatids ; growth & development ; Testis ; cytology ; transplantation ; Transplantation, Heterologous

OBJECTIVETo develop the mechanism of improving protection function of prepared Cornus officinalis for liver and kidney and the biological activity of 5-hydroxymethylfurfural (5-HMF).

METHODPharmacological and chemical studies were used to choose active part. A compound from active part was separated and appraised. To investigate his biological functions, pharmacological experiment was actualized.

RESULTA component was separated and identified. His is 5-HMF. 5-HMF can protect human vein epidermal cell against H2O2 and glucose and inprove acute liver injury in mice.

CONCLUSION5-HMF is the active component in prepared Cornus officinalis and substance basis for protecting liver and kidney.

Animals ; Cells, Cultured ; Cornus ; chemistry ; Endothelial Cells ; drug effects ; Female ; Furaldehyde ; analogs & derivatives ; chemistry ; isolation & purification ; pharmacology ; Liver ; drug effects ; Male ; Mice ; Random Allocation

Objective To explore the expression of protein arginine methyltransferases 6 (PRMT6) in chronic obstructive pulmonary disease (COPD) mouse model and its correlation with inflam mation gene interleukin 6 (IL-6) and cyclooxygenase 2 (COX-2).Methods Sixteen C57BL/8J mice were randomly divided into 2 groups:control group and cigarette smoke extract (CSE)-induced COPD group.Each group was injected intraperitoneally with phosphate-buffered saline solutions (PBS) or CSE at days 1,12,23 and measured lung function and collected lung tissue at day 29.The morphology change of the lung tissue was determined by hematoxylin and eosin (HE) stainning.The protein expression of PRMT6,H3R2me2a and H3K4me3 were detected in lung homogenates by Western-blotting.The mRNA expression of PRMT6,IL-6 and COX-2 were measured by quantitative real-time polymerase chain reaction (qRT-PCR).Results Comparing to control group,COPD group showed typical emphysema changes in the lung tissue,and significantly decreased lung function.The mRNA and protein expression of PRMT6 in the lung tissue of the mice with COPD were significantly decreased,following with the down-regulated signal level of H3R2me2a protein expression,while the increased level of IL-6 and COX-2 mRNA.Meanwhile,PRMT6 was negatively correlated with IL-6 and COX-2 mRNA expression.Conclusions PRMT6 was significantly reduced in CSE-induced COPD mouse model,following with decreased histone H3R2 dimethylation and increased H3K4 trimethylation,negatively correlating with inflammatory gene IL-6 and COX-2 transcription expression.PRMT6 downregulation may activate the transcriptional expression of inflammatory genes involved in the development of COPD,through the regulation of histone methylation level.

Objective:To analyze the epidemiological characteristics and genotypes of human rhinovirus (HRV) in patients with upper respiratory tract infection in Qingdao in the winter of 2020.Methods:Throat swab samples were collected from 101 patients with upper respiratory tract infection in Qingdao from November 2020 to January 2021. Quantitative PCR was used to detect 15 common respiratory viruses in the samples. HRV-positive samples were further analyzed with RT-PCR to amplify and sequence HRV VP4/VP2 gene. A phylogenetic tree was constructed based on the sequencing results and homology analysis was conducted.Results:Six common respiratory viruses were detected in the 101 patients. Thirty-four cases (34/101, 33.66%) were single pathogen infection and two cases were multiple infection (2/101, 1.98%). The positive rate of HRV was the highest (21.78%, 22/101). Twenty HRV VP4/VP2 sequences were successfully amplified. Phylogenetic analysis showed that there were 16 strains of HRV-A subtype and four strains of HRV-C subtype and 14 serotypes were involved.Conclusions:HRV was one of the leading viral pathogens causing upper respiratory tract infection in Qingdao in the winter of 2020 and the predominant subtype was HRV-A.

OBJECTIVETo explore the role of NF-E2-related factor 2(Nrf2) and its related factors in the progression of nonalcoholi steatohepatitis (NASH) by investigating the alterations of lipid metabolism and liver histopathology as well as the changes of mRNA and protein expression levels of Nrf2 and its related factors in rats during NASH progression.

METHODSMale SD rats were randomly divided into normal group and model group, which were administrated with high fat diet to establish nonalcoholic steatohepatitis model. The rats from both groups were randomly killed at the end of 4, 12 weeks respectively. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) were detected in the serum and liver tissue; Changes in fat deposition in liver tissue were determined by oil red O staining. HE staining were used to observe the pathological changes of liver tissue and to calculate nonalcoholic fatty liver disease (NAFLD) activity score (hepatic steatosis, inflammation and ballooning degeneration of liver cells). The expression of Nrf2 in liver was detected by immunohistochemical staining. The mRNA and protein levels of Nrf2 and related factors in liver were determined by Realtime PCR and Western blot, respectively.

RESULTSAfter 4 weeks of high fat diet, the levels of ALT, AST, TC in rat serum and TC, TG, LDL-C in liver were significantly increased compared with that of the normal group (P < 0.01, P < 0.05). After 4 weeks of high fat diet, the levels of ALT, AST, TC, TG in serum and TC, TG, LDL- C in liver increased further (P < 0.01, P < 0.05). Until the 12th week, the content of HDL-C in liver was significantly lower than that of the normal group (P < 0.05). At the end of the 4th or the 12th week, lipid droplets in the model rat liver cells were heavily dyed red and hepatic steatosis increased severely, with ballooning degeneration of liver cells. With the extension of high fat diet feeding time, fat deposition in the liver tissue, hepatic steatosis, NAFLD score, Nrl2 expression were significantly increased (P < 0.01). Expression levels of mRNA and protein of Nrf2, heme oxyenase 1(HO1), NADPH quinone oxidoreductase 1(NQO1), γ-glutamylcysteine synthethase (γ-GCS), glutathione S-transferase (GST) in the model rats increased or decreased at the end of the 4th or the 12th week differentially, (P < 0.01, P < 0.05) with the more significant changes at the end of the 4th week than the 12th week.

CONCLUSIONNrf2 and its related factors may be involved in the occurrence and development of nonalcoholic fatty liver disease, which may play an important role in the process of NASH formation.

Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; metabolism ; Cholesterol ; metabolism ; Diet, High-Fat ; Dipeptides ; metabolism ; Disease Progression ; Glutathione Transferase ; metabolism ; Heme Oxygenase (Decyclizing) ; metabolism ; Lipid Metabolism ; Liver ; pathology ; Male ; NAD(P)H Dehydrogenase (Quinone) ; metabolism ; NF-E2-Related Factor 2 ; metabolism ; Non-alcoholic Fatty Liver Disease ; metabolism ; Rats ; Rats, Sprague-Dawley ; Triglycerides ; metabolism

OBJECTIVETo study the alterations and relationship of surfactant protein (SP)-A, SP-D and KL-6 in serum and bronchoalveolar lavage fluids (BALF) in children with Mycoplasma pneumoniae pneumonia (MPP).

METHODSelf-control method was used for the study on SP-A, SP-D and KL-6 in serum, infected and non-infected BALFs in 32 MMP children with only one side of MPP.

RESULTThe contents of SP-A, SP-D and KL-6 in infected BALF were [mg/L;M (IQR) ]: 243 (90-468) , 187 (43-333) , 148 (47-426) ;104 (37-257) , 56 (25-131) , 35 (12-147) in non-infected BALF; 35 (25-69) , 33 (9-149) and 24 (15-62) in serum. The correlation coefficient of KL-6 between serum and infected BALF were -0.534 and -0.378 (P < 0.05).

CONCLUSIONThere were significant correlation between the alterations of SP-A, SP-D and KL-6 in serum and lung infection in children with CAP. KL-6 in serum may be more sensitive than SP-A and SP-D.

Adolescent ; Biomarkers ; blood ; metabolism ; Bronchoalveolar Lavage Fluid ; chemistry ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Lung ; metabolism ; pathology ; Male ; Mucin-1 ; blood ; metabolism ; Pneumonia, Mycoplasma ; blood ; metabolism ; Pulmonary Surfactant-Associated Protein A ; blood ; metabolism ; Pulmonary Surfactant-Associated Protein D ; blood ; metabolism ; Severity of Illness Index

BACKGROUNDTransforming growth factor-β1 (TGF-β1) is known to have a role in keloid formation through the activation of fibroblasts and the acceleration of collagen deposition. The objective of this current study was to isolate TGF-β1 phage model peptides from a phage display 7-mer peptide library to evaluate their therapeutic effect on inhibiting the activity of keloid fibroblasts.

METHODSA phage display 7-mer peptide library was screened using monoclonal anti-human TGF-β1 as the target to obtain specific phages containing ectogenous model peptides similar to TGF-β1. Enzyme-linked immunosorbent assay (ELISA) was performed to select monoclonal phages with good binding activity, which underwent DNA sequencing. MTT assay and apoptosis assessment were used to evaluate the biological effects of the phage model peptides on keloid fibroblasts. Immunofluorescence assay was employed to show the binding affinity of the model peptides on phages causing keloid fibroblasts. Quantitative real-time PCR analysis was carried out to detect the expressions of nuclear factor κB (NF-κB) mRNA, connective tissue growth factor (CTGF) mRNA and TGF-β receptor II (TβRII) mRNA in keloid fibroblasts.

RESULTSSpecific phages with good results of ELISA were beneficiated. Four phage model peptides were obtained. The data of MTT showed that TGF-β1 and one phage model peptide (No. 4) could promote keloid fibroblasts proliferation, however, three phage model peptides (No. 1 - 3) could inhibit keloid fibroblasts proliferation. The results of apoptosis assessment showed that the three phage model peptides could slightly induce the apoptosis in keloid fibroblasts. The data of immunofluorescence assay revealed that the model peptides on phages rather than phages could bind to keloid fibroblasts. The findings of quantitative real-time PCR analysis suggested that the expressions of NF-κB mRNA and CTGF mRNA in the three phage model peptide groups decreased, while the expression of TβRII mRNA slightly increased.

CONCLUSIONSThree phage model peptides isolated from a phage display 7-mer peptide library can inhibit keloid fibroblasts proliferation and induce the apoptosis in keloid fibroblasts. They can inhibit the activity of keloid fibroblasts by blocking TGF-β1 binding to its receptor and then regulating the expressions of NF-κB, CTGF and TβRII.

Apoptosis ; Cell Line ; Cell Proliferation ; drug effects ; Enzyme-Linked Immunosorbent Assay ; Fibroblasts ; cytology ; drug effects ; Fluorescent Antibody Technique ; Humans ; Peptide Library ; Peptides ; immunology ; pharmacology ; Polymerase Chain Reaction ; Transforming Growth Factor beta1 ; immunology

Objective To explore the value of patient global assessment (PGA) on evaluating disease activity in patients with axial spondyloarthritis (SpA),Methods A total of 222 patients with axial SpA were recruited.Scores of PGA,disease activity index [Bath ankylosing spondylitis disease activity index (BASDAI),ankylosing spondylitis disease activity score (ASDAS)crp] and spondyloarthritis research consortium of Canada (SPARCC) were calculated.Differences of PGA scores between different disease activity groups in axial SpA were compared and correlations between different disease activity index with PGA scores were analyzed.Statistical analyses were performed using Statistical Product and Service Solutions (SPSS) software (version 17.0).Comparison of frequency among different groups was performed by x2 test.Rank-sum test was used to compare the median of measurement data in different groups when the data were skewed in distribution.Cut-off value of PGA for assessing disease activity in axial SpA was calculated by ROC curve.Results Medians of PGA score in groups with BASDAI remission[3(1,4) vs 5(4,7)] and ASDAScrp remission [1(1,2) vs 4(2,5)] were lower than that in disease activity group (P＜0.01).BASDAI scores [1.80(1.20,2.90) vs 3.40(2.28,4.63) vs 5.15 (4.08,5.88)] and ASDAScrp scores [2.19(1.34,2.76) vs 2.86(2.08,3.54) vs 4.08(2.96,4.41)] were significant different among PGA groups (≤3,4-6 and ≥7) (P＜0.01).Differences of SPARCC scores [6.00(0,18.00) vs 7.50(3.75,18.00) vs 18.50(6.75,24.50)] were statistically significant among PGA groups (Z=7.427,P=0.037).Erythrocyte sedimentation rate (ESR) [12.00(5.00,23.00) mm/1 h vs 19.50(7.00,44.50) mm/1 h vs 18.00(7.75,54.75) mm/1 h],C-reactive protein (CRP) [7.85(2.37,22.49) mg/L vs 10.07(3.02,28.51) mg/L vs 21.28(7.14,37.74) mg/L] and Bath ankylosing spondylitis functional index (BASFI) [0.70(0.10,1.30) vs 2.25(0.60,3.30) vs 2.85(0.83,6.53)] were also different among PGA groups (P＜0.01,separately).Proportion of axial SpA patients in BASDAI disease activity group or ASDAScrp higher disease activity group were different among PGA groups (P＜0.01,separately),while represented as positive correlations (P＜0.01,separately).Correlation analyses revealed that PGA was positively correlated with ASDAScrp (r=0.694),BASDAI(r=0.616),SPARCC (r=0.271),ESR (r=0.288),CRP(r=0.215),occipital wall distance (r=0.196),finger-floor distance (r=0.385) and negatively correlated with Sschober's test (r=-0.195) (P＜0.05).Receiver operator characteristic (ROC) curve analysis found that PGA-BASDAI AUC was 0.813,the cut off value of PGA was 3.5 and PGA-ASDAScrp AUC was 0.860,the cut off value of PGA was 2.5.Conclusion PGA has good correlations with the disease activity indexes in axial SpA patients.It can also reflect the degree of inflammation in iconography.PGA may reflect disease activity especially when the value of PGA is around 3.

BACKGROUNDKeratinocyte growth factor (KGF) significantly influences epithelial wound healing. The aim of this study was to isolate KGF phage model peptides from a phage display 7-mer peptide library to evaluate their effect on promoting epidermal cell proliferation.

METHODSA phage display 7-mer peptide library was screened using monoclonal anti-human KGF antibody as the target. Enzyme linked immunosorbent assay (ELISA) was performed to select monoclonal phages with good binding activity. DNA sequencing was done to find the similarities of model peptides. Three-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, immunofluorescence assay and quantitative real-time PCR analysis were employed to evaluate the effect of the phage model peptides on epidermal cells.

RESULTSThirty-three out of fifty-eight (56.9%) of the isolated monoclonal phages exhibited high binding activity by ELISA. Ten of fifteen obtained phage model peptides were similar to KGF or epidermal growth factor (EGF). MTT assay data showed that four (No. 1 - 4) of the ten phage model peptides could promote epidermal cell proliferation. The expression of keratinocyte growth factor receptor (KGFR) mRNA in the KGF control group and the two phage model peptide groups (No. 1 and No. 2) increased. Expression of c-Fos mRNA and c-Jun mRNA in the KGF control group increased, but did not increase in the four phage model peptide groups (No.1 - 4).

CONCLUSIONFour phage model peptides isolated from the phage display 7-mer peptide library can safely promote epidermal cell proliferation without tumorigenic effect.

Cell Proliferation ; drug effects ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Epidermis ; cytology ; Fibroblast Growth Factor 7 ; chemistry ; pharmacology ; Humans ; Peptide Library ; Peptides ; chemistry ; pharmacology ; Polymerase Chain Reaction ; Receptor, Fibroblast Growth Factor, Type 2 ; genetics

A patient suffered a sustained soft tissue necrosis and infection at the radial interphalangeal joint of left thumb after laser nevus removal. He was treated in the Department of Orthopaedics, No. 920 Hospital of Joint Logistic Support Force of Chinese People's Liberation Army in February 2020. CTA combined with digital technology of Mimics software was used to accurately locate the perforator of posterior tibial artery septal perforator flap at the appropriate part of the calf and the super flap (1.20 cm×0.80 cm×0.46 cm) for the repair was designed. After 1 year of follow-up, the left thumb flap had no swelling with a satisfactory texture and appearance. The sensory recovered to S 3, and the left thumb movement was completely normal. Only a linear scar remained at the donor site of the calf.