Humans ; Mpox (monkeypox) ; China

Objective: To investigate the influence of reactive oxygen species (ROS) responsive self-assembled nanomicelle loaded with pyroptosis inhibitor on full-thickness skin defects in diabetic rats. Methods: Experimental research methods were employed. A nucleotide-binding oligomerization domain (NOD) 1/2 inhibitor (NOD-IN-1) was encapsulated with nanomicelle polyethylene glycol-block-polypropylene sulfide (PEG-b-PPS), and the resulting product was called PEPS@NOD-IN-1. The morphology and hydration particle size of PEG-b-PPS and PEPS@NOD-IN-1 were observed by transmission electron microscope and particle size analyzer, respectively, and the encapsulation rate and drug loading rate of PEPS@NOD-IN-1 to NOD-IN-1 and the cumulative release rate of NOD-IN-1 by PEPS@NOD-IN-1 in phosphate buffer solution (PBS) alone and hydrogen peroxide-containing PBS within 40 h were measured and calculated by microplate reader, and the sample number was 3. Twenty-four male Sprague-Dawley rats aged 6-7 weeks were injected with streptozotocin to induce type 1 diabetes mellitus. Six full-thickness skin defect wounds were made on the back of each rat. The injured rats were divided into PBS group, NOD-IN-1 group, PEG-b-PPS group, and PEPS@NOD-IN-1 group with corresponding treatment according to the random number table, with 6 rats in each group. The wound healing was observed on post injury day (PID) 3, 7, and 12, and the wound healing rate was calculated. The ROS levels in wound tissue were detected by immunofluorescence method on PID 3. On PID 7, the granulation tissue thickness in wound was assessed by hematoxylin-eosin staining, the mRNA expressions of NOD1 and NOD2 were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction, and the protein expressions of NOD1, NOD2, and GSDMD-N terminals were detected by Western blotting. Six wounds from different rats in each group were taken for detection of the above indicators. Wound tissue (3 samples per group) was taken from rats in PBS group and PEPS@NOD-IN-1 group on PID 7, and transcriptome sequencing was performed using high-throughput sequencing technology platform. Differentially expressed genes (DEGs) significantly down-regulated in PEPS@NOD-IN-1 group as compared with PBS group were screened, and the enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) was performed. The DEG heatmap of the NOD-like receptor pathway, a pyroptosis-related pathway, was made. Protein-protein interaction (PPI) analysis of DEGs in heatmap was performed through the STRING database to screen key genes of PEPS@NOD-IN-1 regulating the NOD-like receptor pathway. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and Tukey test. Results: PEG-b-PPS and PEPS@NOD-IN-1 were in spherical structures of uniform size, with hydration particle sizes of (134.2±3.3) and (143.1±2.3) nm, respectively. The encapsulation rate of PEPS@NOD-IN-1 to NOD-IN-1 was (60±5)%, and the drug loading rate was (15±3)%. The release of NOD-IN-1 from PEPS@NOD-IN-1 in PBS alone was slow, and the cumulative release rate at 40 h was only (12.4±2.3)%. The release of NOD-IN-1 from PEPS@NOD-IN-1 in hydrogen peroxide-containing PBS within 10 h was very rapid, and the cumulative release rate at 10 h reached (90.1±3.6)%. On PID 3 and 7, the wounds of rats in the four groups were gradually healed, and the healing in PEPS@NOD-IN-1 group was better than that in the other three groups. On PID 12, the wound scab area in PBS group was large, the wound epithelialization in NOD-IN-1 group and PEG-b-PPS group was obvious, and the wound in PEPS@NOD-IN-1 group was close to complete epithelialization. Compared with those in PBS group, NOD-IN-1 group, and PEG-b-PPS group, the wound healing rates on PID 7 and 12 in PEPS@NOD-IN-1 group were significantly increased (P<0.05), the level of ROS in wound tissue on PID 3 was significantly decreased (P<0.05), the thickness of granulation tissue in wound on PID 7 was significantly thickened (P<0.05), and the mRNA expressions of NOD1 and NOD2 and the protein expressions of NOD1, NOD2, and GSDMD-N terminals in wound tissue on PID 7 were significantly decreased (P<0.05). KEGG pathway analysis showed that DEGs significantly down-regulated in PEPS@NOD-IN-1 group as compared with PBS group were significantly enriched in NOD-like receptors, hypoxia-inducible factors, mitogen-activated protein kinases, and tumor necrosis factor (TNF) pathways. In the DEG heatmap of NOD-like receptor pathway, the genes regulating pyroptosis mainly involved NOD1, NOD2, NOD-like receptor thermoprotein domain-related protein 3, Jun, signal transduction and transcriptional activator 1 (STAT1), TNF-α-induced protein 3. The PPI results showed that NOD1, NOD2, and STAT1 were the key genes of PEPS@NOD-IN-1 regulating the NOD-like receptor pathway. Conclusions: PEPS@NOD-IN-1 can down-regulate the level of local ROS in wounds and the expression of NOD1, NOD2, and GSDMD-N terminals, the key regulators of pyroptosis, thereby promoting the repair of full-thickness skin defect wounds in diabetic rats. PEPS@NOD-IN-1 can also significantly down-regulate the pyroptosis, inflammation, and hypoxia-related pathways of wounds, and regulate NOD-like receptor pathways by down-regulating key genes NOD1, NOD2, and STAT1.
Rats ; Male ; Animals ; Reactive Oxygen Species ; Wound Healing ; Rats, Sprague-Dawley ; Diabetes Mellitus, Experimental ; Hydrogen Peroxide ; Pyroptosis ; Skin Abnormalities ; Soft Tissue Injuries ; NLR Proteins ; Hypoxia ; RNA, Messenger

A 36-year-old woman was admitted to the Peking Union Medical College Hospital with a history of fractures for 2 years, limb weakness for 1 year, and ostealgia for 2 months. The patient's examination identified iron deficiency anemia, significantly decreased serum calcium and 25-hydroxyvitamin D3 levels, and increased alkaline phosphatase and parathyroid hormone levels. Imaging showed several typical signs of osteomalacia. Considering the history of Roux-en-Y gastric bypass surgery, the diagnosis was considered to be osteomalacia caused by a postoperative nutritional absorption disorder. The patient was supplemented with calcitriol, calcium, and vitamin D and gradually returned to normal physical activity. The bone metabolism indicators and bone density were significantly improved.

Objective: To explore the heterogeneity and correlation of clinical phenotypes and genotypes in children with disorders of sex development (DSD). Methods: A retrospective study of 1 235 patients with clinically proposed DSD in 36 pediatric medical institutions across the country from January 2017 to May 2021. After capturing 277 DSD-related candidate genes, second-generation sequencing was performed to analyzed the heterogeneity and correlation combined with clinical phenotypes. Results: Among 1 235 children with clinically proposed DSD, 980 were males and 255 were females of social gender at the time of initial diagnosis with the age ranged from 1 day of age to 17.92 years. A total of 443 children with pathogenic variants were detected through molecular genetic studies, with a positive detection rate of 35.9%. The most common clinical phenotypes were micropenis (455 cases), hypospadias (321 cases), and cryptorchidism (172 cases) and common mutations detected were in SRD5A2 gene (80 cases), AR gene (53 cases) and CYP21A2 gene (44 cases). Among them, the SRD5A2 mutation is the most common in children with simple micropenis and simple hypospadias, while the AMH mutation is the most common in children with simple cryptorchidism. Conclusions: The SRD5A2 mutation is the most common genetic variant in Chinese children with DSD, and micropenis, cryptorchidism, and hypospadias are the most common clinical phenotypes. Molecular diagnosis can provide clues about the biological basis of DSD, and can also guide clinicians to perform specific clinical examinations. Target sequence capture probes and next-generation sequencing technology can provide effective and economical genetic diagnosis for children with DSD.
3-Oxo-5-alpha-Steroid 4-Dehydrogenase/genetics* ; Child ; China/epidemiology* ; Cryptorchidism/genetics* ; Disorders of Sex Development/genetics* ; Female ; Genital Diseases, Male ; Genotype ; Humans ; Hypospadias/genetics* ; Male ; Membrane Proteins/genetics* ; Penis/abnormalities* ; Phenotype ; Retrospective Studies ; Steroid 21-Hydroxylase/genetics*

Objective To evaluate whether the patients with Parkinson′s Disease (PD) having higher occurrence rate of frac-ture .Methods CMB ,CNKI ,PubMed ,Embase ,Web of Science ,Medline ,Embase and Cochrane Library were retrieved ,meanwhile which was assisted by the manual retrieval .The retrieval time was until February 2017 .The cohort studies on the occurrence rate of fracture in PD patients were collected .Then the included studies were analyzed after the data extraction and treatment evaluation . Results A total of 1160 articles were retrieved ,finally 11 cohort studies were included ,involving 988723 subjects .The analysis showed that the fracture occurrence risk in PD patients was significantly higher than that in the control group (RR=2 .09 ,95% CI:1 .91-2 .28) ,in which the occurrence rate of hip fracture was significantly higher than that in the control group (RR=2 .33 ,95%CI:1 .79-3 .02) ,while the occurrence rate of spinal fracture had no statistical difference (RR=1 .33 ,95% CI:0 .78-2 .27) ,and the fracture occurrence risk in male and female patients of PD group was significantly higher than that in the control group (RR=2 .40 , 1 .69 ,95% CI:2 .21-2 .60 ,1 .62-1 .76) .Conclusion The fracture occurrence risk in PD patients is significantly higher than that in the control group ,due to existence of certain geographic bias and publication bias risk ,it is needed more high quality clinical stud-ies to accurately evaluate whether the fracture risk in PD patients being much higher .

Objective To investigate the relationship between geranylgeranyl pyrophosphate synthase (GGPPS) gene polymorphisms and bone response to alendronate in Chinese osteoporotic women.Methods A total of 639 postmenopausal women with osteoporosis or osteopenia were included and randomly received treatment of low dose (70 mg per two weeks) or standard dose (70 mg weekly) of alendronate for one year. The six tag single nucleotide polymorphisms of GGPPS gene were identified. Bone mineral density (BMD), serum cross-linked C-telopeptide of type I collagen (β-CTX), and total alkaline phosphatase (ALP) were measured before and after treatment. GGPPS gene polymorphisms and the changes of BMD and bone turnover markers after treatment were analyzed.Results rs10925503 polymorphism of GGPPS gene was correlated to serum β-CTX levels at baseline, and patients with TT genotype had significantly higher serum β-CTX level than those with TC or CC genotype (all P<0.05). No correlation was found between polymorphisms of GGPPS gene and serum total ALP levels, as well as BMD at baseline. After 12 months of treatment, lumbar spine and hip BMD increased and serum bone turnover markers decreased significantly (P<0.01), and without obvious differences between the low dose and standard dose groups (all P>0.05). However, GGPPS gene polymorphisms were uncorrelated to percentage changes of BMD, serum total ALP, and β-CTX levels (all P>0.05).Conclusion GGPPS gene polymorphisms are correlated to osteoclasts activity, but all tag single nucleotide polymorphisms of GGPPS gene have no influence on the skeletal response to alendronate treatment.

Objective To investigate the phenotypes of a kindred with progressive diaphyseal dysplasia ( PDD) and to detect the mutation of transforming growth factor beta-1 ( TGFB1 ) gene. Methods A PDD pa-tient of a non-consanguineous family presented with early onset in childhood, who suffered from lower limb pain, fatigability and muscle weakness. Her clinical manifestations, features of skeletal X-ray examination, and bone turnover markers were evaluated. Mutation of TGFB1 was identified by direct Sanger sequencing of polymerase chain reaction amplification product. Results The proband presented with elevated bone turnover biomarkers, and nonuniform thickening and sclerosis of bone cortex of limbs in X-ray films. A heterozygous missense mutation c. 652C>T (p. Arg218Cys) in exon 4 of TGFB1 was identified in the proband, but not in either of her par-ents. Glucocorticoid was given and after 4 months of treatment, the bone pain and activity were obviously im-proved. Conclusions The typical clinical manifestations of PDD are limb pain and diaphyseal hyperostosis. The missense mutations at position 218 of TGFB1 are hotspot pathogenic mutations of PDD. Glucocorticoids can miti-gate the symptoms in PDD patients.

Objective To understand molecular characteristics of Japanese encephalitis virus ( JEV ) isolated from the major Japanese encephalitis epidemic areas in Sichuan Province,and to provide the foundation for JEV prevention.Methods 13 JEV strains were isolated from mosquitoes in Sichuan during 2007-2010,E genes and preM genes were sequenced and phylogenetic analyses were performed using MEGA5 molecular software.Results Phylogeoetic analysis indicated that all 13 JEV strains from Sichuan belonged to genotype Ⅰ,homologies at nucleotide level and deduced amino acid level in PreM gene were 97％-100％ and 98.7％-100％,and 97.8％-99.9％ and 99.6％-100％ in E gene,respectively.Homologies at nucleotide level and deduced amino acid level in PreM gene between 13 JEV strains and JEV isolated in 2004 in Sichuan were 96.2％-99.1％ and 97.5％-98.7％,and were 97.7％-99.6％ and 98.6％-100％ in E gene,respectively.By comparison with vaacine strains P3 and SA14-14-2,homologies at nucleotide level and deduced amino acid level were 84.1％-85.8％ and 93.7％-96.2％ in PreM gene,and were 87.6％-88.3％ and 97％-97.8％ in E gene,respectively.The neurovirulence-related 8 amino acid sites encode by E gene remained unchanged in 13 JEV strains.Conclusion JEV with genotype Ⅰ predominated in Sichuan,nucleotide sequences and deduced amino acid sequences in PreM gene and E gene were highly conserved,key neurovirulence-rerlated sites remained unchanged.It suggested currently used vaccine is still capable of preventing JEV infection.

Objective To explore the effect of clinical path on ache nursing of malignant tumor patients. Methods 100 tumor patients received interventional therapy was divided into observation group and control group. The observation group was treated with self-made clinical path nursing, and the control group was treated with normal nursing, the compliance of nursing and analgesic effect was compared between the two groups. Results Ache expression, medicine taken on time, addiction cognition and medicine side effect understanding between the two groups were significantly different (P <0. 01 ). The analgesic effect was also different between the two groups ( P < 0. 05). Conclusions Application of clinical path nursing on patients carried interventional therapy could enhance the compliance of patients, improve the relationship between the nurses and the patients, and increase the satisfactory degree.

AIMTo study the metabolic pathways of ginsenoside Rd in rats.

METHODSUrine samples were collected before and after 24 h of single oral administration of 150 mg and intravenous administration of 60 mg of ginsenoside Rd to six rats, separately. The samples were purified by SPE column and then were analyzed by liquid chromatography-ESI-mass spectrometry for putative metabolites.

RESULTSParent drug and its seven metabolites were identified in rat urine based on comparing total ion chromatograms of the blank with the metalolic urine as well as mass spectra. Its main metabolic pathways and possible structures are elucidated.

CONCLUSIONOxidation, combination and deglucosylation were found to be the major metabolic pathway of ginsenoside Rd in rats.

Administration, Oral ; Animals ; Chromatography, High Pressure Liquid ; methods ; Ginsenosides ; administration & dosage ; metabolism ; urine ; Injections, Intravenous ; Male ; Oxidation-Reduction ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Spectrometry, Mass, Electrospray Ionization ; methods