OBJECTIVE To investigate the potential mechanism by which Juanxiao decoction improves bronchial asthma （hereinafter referred to as “asthma”） based on the nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 （NLRP3） inflammasome signaling pathway. METHODS Female SD rats were randomly assigned to normal group， model group and Juanxiao decoction low-， medium- and high-dose groups （0.36， 0.72 and 1.44 g/kg， calculated based on crude drug weight）， as well as positive control group （Dexamethasone acetate tablets， 0.2 mg/kg）， with 10 rats in each group. Except for the normal group， asthma models were established in the remaining groups via intraperitoneal injection of ovalbumin combined with aluminum hydroxide， followed by nebulized inhalation of ovalbumin. On day 14 of the experiment， rats in each group received intragastric administration of the corresponding solution or normal saline， once a day， for 7 consecutive days. Following the final administration， the following parameters were measured in each group： lung function indexes （forced vital capacity， forced expiratory volume in 0.3 second， peak expiratory flow）， serum levels of inflammatory markers （interleukin-1β， interleukin- 18）， and the percentages of inflammatory cells （lymphocytes， eosinophils， neutrophils） in bronchoalveolar lavage fluid. Histopathological changes in lung tissue were observed， and the protein and mRNA expressions of nuclear factor-kappa B （NF- κB）， NLRP3 and caspase-1 in lung tissue were detected. RESULTS Compared with the normal group， pathological changes such as alveolar wall thickening and inflammatory cell infiltration were observed in rats in the model group. All pulmonary function indicators were significantly reduced in rats in the model group and the administration groups. The levels of inflammatory markers， the percentages of inflammatory cells， and the protein and mRNA expressions of NF-κB， NLRP3 and caspase-1 were significantly elevated or up-regulated （P＜0.05）. Compared with the model group， pathological changes in rats in each dosage group of Juanxiao decoction were significantly alleviated， and all quantitative indicators showed dose-dependent improvements （P＜0.05）. CONCLUSIONS Juanxiao decoction can reduce airway inflammatory responses in asthmatic rats， alleviate lung function impairment， and improve pathological changes such as inflammatory cell infiltration. Those effects may be related to the inhibition of the NLRP3 inflammasome signaling pathway.

Objective To investigate the composition of symptom clusters in early postoperative breast cancer patients and analyze the sentinel symptoms of each cluster of symptoms. Methods A total of 309 patients who underwent mastectomy were conveniently sampled and surveyed using the Chinese version of the Anderson Symptom Inventory. Principal component analysis and varimax orthogonal rotation were employed to analyze the symptom clusters, and their associations were analyzed using the Apriori algorithm model to identify the sentinel symptoms of each cluster of symptoms. Results Three symptom clusters were identified in early postoperative breast cancer patients: neuro-sleep symptom cluster [fatigue (weakness)-distress-pain-sleepiness-restless sleep], sensory-perception symptom cluster (numbness-forgetfulness-shortness of breath-sadness-dry mouth), and digestive system symptom cluster (nausea-vomiting-loss of appetite). Fatigue was the sentinel symptom of the neuro-sleep symptom cluster, numbness was the sentinel symptom of the sensory-perception symptom cluster, and nausea was the sentinel symptom of the digestive system symptom cluster. Conclusion Early postoperative breast cancer patients experience multiple symptom clusters, with sentinel symptoms existing in each cluster. Healthcare staff should develop intervention measures based on sentinel symptoms to improve the efficiency of symptom management and reduce the degree of symptom distress for patients.

OBJECTIVE: To detect variants of IVD gene among 4 neonates with suspected isovalerate acidemia in order to provide a guidance for clinical treatment. METHODS: 111 986 newborns and 7461 hospitalized children with suspected metabolic disorders were screened for acyl carnitine by tandem mass spectrometry. Those showing a significant increase in serum isovaleryl carnitine (C5) were analyzed for urinary organic acid and variants of the IVD gene. RESULTS: Four cases of isovalerate acidemia were detected, which included 2 asymptomatic newborns (0.018‰, 2/111 986) and 2 children suspected for metabolic genetic diseases (0.268‰, 2/7461). The formers had no obvious clinical symptoms. Analysis of acyl carnitine has suggested a significant increase in C5, and urinary organic acid analysis has shown an increase in isovaleryl glycine and 3-hydroxyisovalerate. Laboratory tests of the two hospitalized children revealed high blood ammonia, hyperglycemia, decreased red blood cells, white blood cells, platelets and metabolic acidosis. The main clinical manifestations have included sweaty foot-like odor, feeding difficulty, confusion, drowsiness, and coma. Eight variants (5 types) were detected, which included c.158G>A (p.Arg53His), c.214G>A (p.Asp72Asn), c.548C>T (p.Ala183Val), c.757A>G (p.Thr253Ala) and 1208A>G (p.Tyr403Cys). Among these, c.548C>T and c.757A>G were unreported previously. None of the variants was detected by next generation sequencing of 2095 healthy newborns, and all variants were predicted to be likely pathogenic based on the guidelines from the American College of Medical Genetics and Genomics. CONCLUSION The incidence of isovalerate acidemia in Liuzhou area is quite high. Screening of metabolic genetic diseases is therefore recommended for newborns with abnormal metabolism. The discovery of novel variants has enriched the mutational spectrum of the IVD gene.
Infant, Newborn ; Child ; Humans ; Acidosis ; Carnitine ; Erythrocytes ; High-Throughput Nucleotide Sequencing

Although widely applied in treating hematopoietic malignancies, transplantation of hematopoietic stem/progenitor cells (HSPCs) is impeded by HSPC shortage. Whether circulating HSPCs (cHSPCs) in steady-state blood could be used as an alternative source remains largely elusive. Here we develop a three-dimensional culture system (3DCS) including arginine, glycine, aspartate, and a series of factors. Fourteen-day culture of peripheral blood mononuclear cells (PBMNCs) in 3DCS led to 125- and 70-fold increase of the frequency and number of CD34⁺ cells. Further, 3DCS-expanded cHSPCs exhibited the similar reconstitution rate compared to CD34⁺ HSPCs in bone marrow. Mechanistically, 3DCS fabricated an immunomodulatory niche, secreting cytokines as TNF to support cHSPC survival and proliferation. Finally, 3DCS could also promote the expansion of cHSPCs in patients who failed in HSPC mobilization. Our 3DCS successfully expands rare cHSPCs, providing an alternative source for the HSPC therapy, particularly for the patients/donors who have failed in HSPC mobilization.
Antigens, CD34/metabolism* ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; Humans ; Leukocytes, Mononuclear/metabolism* ; Peptides/metabolism*

Objective To study the effect of pirfenidone (PFD) on the transformation of rat corneal stromal cells into fibroblasts in vitro and further explore the anti-fibrotic effect of PFD. Methods The corneal stromal cells from SD rat was isolated and cultured ,and was determined by vimentin stain. The experiment was divided into control group(DMEM+10%FBS),TGF-β1 group(2 ng/mL TGF-β1+DMEM+10%FBS)and PFD group(1 mg/mL PFD+ 2 ng/mL TGF-β1+DMEM+ 10%FBS). Cell proliferation was detected by CCK-8 assay. Collagen Ⅰ,Collagen Ⅲ,Keratocan and CD99 expression were detected by Western blot. Results Compared with control group and TGF-β1 group,the cell proliferation were significantly decreased in PFD group(P<0.05). Western blot showed that PFD can up-regulated Collagen Ⅰand Keratocan but down-regulated Collagen ⅢCD90 expression(P < 0.05). The ratio of Collagen Ⅲ/Collagen Ⅰ in PFD group was lowest in all groups(P < 0.05). Conclusion PFD can resistant fibration in corneal stromal cells may through the inhibition of TGF-β ,which affect the collagen synthesis and Keratocan,CD90 expression.

Objective To investigate the structure and mechanical properties of pods after dehydration and the biomechanical mechanism of spreading pod seed injection due to torsion crack. Methods The layered pods, the cell size and direction at different cellular layers were analyzed by histology, microstructure observation, mechanical property test and high-speed photography. The process of pod ejection was observed, and the principle of pod ejection was summarized. Results The ejection of pods started from the crack of the bottom, and cracked gradually from the bottom to the top. The cell arrangement of two parts of the same pod was opposite. Each pod was divided into 4 layers wherein the first exterior layer and the middle layer were orthogonal to each other. There was a layer of cells between the first exterior layer and the middle layer, of which the cell wall was broken. In the process of dehydration, fibers in the outer layer shrank and fibers in the middle layer stretched. Conclusions Pod fiber will be contracted in the orthogonal direction after dehydration to accumulate elastic performance and generate pre-stress, and finally the pod is cracked to release the pre-stress.

Objective To analyze the genotype and plasmid transfer of Enterobacteriaceae carring blaNDM‐1 with blaIMP‐4 or blaKPC‐2 .Methods From April 2012 to October 2014 ,a total of 33 non‐repeatitive carbapenem‐resistant Enterobacteriaceae ( including Imipenem‐resistant , meropenem‐resistant or Ertapenem‐resistant) were isolated from 5 hospitals in Wenzhou and Hangzhou . Identification and antimicrobial susceptibility test were performed using Vitek 2 Compact automatic microbiology analyzer . Phenotypes of carbapenemase were screened using modified Hodge test and ethylenediamine tetraacetic acid‐disk synergy test .Extended spectrum βlactamase test was determined by the double disk combination test which was recommended by Clinical and Laboratory Standards Institute .AmpC activity was tested by a three‐dimensional Cefoxitin method .Drug resistant genes including blaNDM‐1 and linkage of ISAba125‐NDM were detected by polymerase chain reaction (PCR) .The purified PCR products were cloned and sequenced .Plasmid conjugation experiment and elimination method were carried out to test partial bacterial strain and K . pneumoniae carrying blaNDM‐1 with blaIMP‐4 or blaKPC‐2 .Results Of the 33 non‐repeatitive carbapenem‐resistant Enterobacteriaceae ,28 were strains of K .pneumoniae ,1 strain of K . oxytoca,2strainsof Escherichiacoli,1strainof K.planticolaand1strainof E.cloacae.Thirteenstrains were isolated from Hospital of Sir Run Run Shaw of Zhejiang University ,thirteen from Wenzhou Hospital of Traditional Chinese Medicine ,one from Wenzhou People′s Hospital ,three from the First Affiliated Hospital of Wenzhou Medical University and three from Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine .Thirty‐one strains were confirmed as carbapenemase‐producing with 24 of blaKPC‐2 ,3 of blaNDM‐1 ,1 of blaNDM‐5 and 3 of blaIMP‐4 .Among them ,one strain carried blaNDM‐1 with blaIMP‐4 and one strain carried blaNDM‐1with blaKPC‐2 ,respectively .The plasmid transfer and conjugation experiment was performed between strains carrying blaNDM‐1 and Escherichia coli EC600 or K . pneumoniae ATCC13833 and genes of blaNDM‐1 and ISAba125‐NDM were obtained .Conclusions blaKPC‐2 gene is the popular carbapenemase genotype .blaNDM‐1 or blaNDM‐5 may be correlated with linkage gene of ISAba125‐N DM .Coexistence of blaNDM‐1 carrying blaIMP‐4 or blaKPC‐2 is detected in the same strain , respectively . Enough importance should be attached to the strains ,because most of them are multiple drug resistance with related genes located in the plasmid which is easily spread between strains .