BACKGROUND: Previous research demonstrated that a homozygous mutation of g.136372044G>A (S12N) in caspase recruitment domain family member 9 ( CARD9 ) is critical for producing Aspergillus fumigatus -induced ( Af -induced) T helper 2 (T H 2)-mediated responses in allergic bronchopulmonary aspergillosis (ABPA). However, it remains unclear whether the CARD9S12N mutation, especially the heterozygous occurrence, predisposes the host to ABPA. METHODS: A total of 61 ABPA patients and 264 controls (including 156 healthy controls and 108 asthma patients) were recruited for sequencing the CARD9 locus to clarify whether patients with this heterozygous single-nucleotide polymorphisms are predisposed to the development of ABPA. A series of in vivo and in vitro experiments, such as quantitative real-time polymerase chain reaction, flow cytometry, and RNA isolation and quantification, were used to illuminate the involved mechanism of the disease. RESULTS: The presence of the p.S12N mutation was associated with a significant risk of ABPA in ABPA patients when compared with healthy controls and asthma patients, regardless of Aspergillus sensitivity. Relative to healthy controls without relevant allergies, the mutation of p.S12N was associated with a significant risk of ABPA (OR: 2.69 and 4.17 for GA and AA genotypes, P = 0.003 and 0.029, respectively). Compared with patients with asthma, ABPA patients had a significantly higher heterozygous mutation (GA genotype), indicating that p.S12N might be a significant ABPA-susceptibility locus ( aspergillus sensitized asthma: OR: 3.02, P = 0.009; aspergillus unsensitized asthma: OR: 2.94, P = 0.005). The mutant allele was preferentially expressed in ABPA patients with heterozygous CARD9S12N , which contributes to its functional alterations to facilitate Af -induced T H 2-mediated ABPA development. In terms of mechanism, Card9 wild-type ( Card9WT ) expression levels decreased significantly due to Af -induced decay of its messenger RNA compared to the heterozygous Card9S12N . In addition, ABPA patients with heterozygous CARD9S12N had increased Af -induced interleukin-5 production. CONCLUSION Our study provides the genetic evidence showing that the heterozygous mutation of CARD9S12N , followed by allele expression imbalance of CARD9S12N , facilitates the development of ABPA.
Humans ; Aspergillosis, Allergic Bronchopulmonary/complications* ; Aspergillus fumigatus/genetics* ; Asthma/genetics* ; Aspergillus ; Mutation/genetics* ; CARD Signaling Adaptor Proteins/genetics*

OBJECTIVE To explore the potential targets and mechanisms of the modified Baihe dihuang decoction （MBD/ BDD） applied in post-stroke depression （PSD）. METHODS Network pharmacology was used to mine the potential targets and key pathways of MBD/BDD in the treatment of PSD. PSD model rats were induced by focal cerebral ischemia surgery combined with chronic unforeseen mild stress， and then were randomly divided into PSD model group， MBD/BDD group （12.6 g/kg， by raw drug）， and fluoxetine hydrochloride （FLX） group （positive control， 2.3 mg/kg）； a blank control group was also set up， with 8 rats in each group. Each administration group was given a corresponding medication solution by gavage once a day for 21 consecutive days. The intervention effect of MBD/BDD on depression-like symptoms in model rats was evaluated by open field and forced swimming tests. The brain tissues of rats in each group were dissected and total RNA was extracted for transcriptome sequencing and bioinformatics analysis. The mRNA and protein expressions of genes with significant changes and common neurotrophic factors were verified based on the above results. RESULTS A total of 131 MBD/BDD antidepressant-related target genes were obtained （such as IL1B and AKT1， etc.）， which were closely related to neural active ligand-receptor interactions and cyclic adenosine monophosphate signaling pathway. MBD/BDD could significantly prolong or increase the total time spent and distance traveled in the central grid of qiangzhe@cqtcm.edu.cn PSD model rats， and significantly shorten the cumulative immobility time （P＜0.05）. After treatment with MBD/BDD， the number of genes that changed in rat brain tissue was much higher than that in the FLX group， and there were significant differences in gene profiles among the PSD model group， MBD/BDD group， and FLX group. There were 1 351 differentially expressed genes （DEGs） between the MBD/BDD group and the PSD model group， of which 178 were significantly down-regulated and 1 173 were significantly up-regulated （P＜0.05）. Above 1 351 DEGs were involved in neuronal differentiation， chemical synaptic transmission regulation. They were significantly enriched in axonal guidance， cholinergic synapses and neuroactive ligand-receptor interactions. The top 30 genes in terms of up-regulation in the brain tissue of rats of MBD/BDD group were all associated with neuronal proliferation， development， differentiation， and migration. After MBD/BDD intervention， the expressions of Fezf2， Arx， Ostn， Nrgn genes， brain-derived neurotrophic factor and tyrosine kinase receptor B protein in brain tissue of rats were significantly increased （P＜0.05）. CONCLUSIONS The anti-PSD effect of MBD/BDD may be related to the up-regulation of the expression of genes related to neuronal proliferation， development， differentiation and migration， as well as the promotion of neural structural and functional repair.

OBJECTIVES: In this study, we explored how adiponectin mediated urotensin II (UII)‍-induced tumor necrosis factor-‍α (TNF-‍α) and α‍-smooth muscle actin (α‍-SMA) expression and ensuing intracellular signaling pathways in adventitial fibroblasts (AFs). METHODS: Growth-arrested AFs and rat tunica adventitia of vessels were incubated with UII and inhibitors of signal transduction pathways for 1‍‒‍24 h. The cells were then harvested for TNF-α receptor (TNF-‍α-R) messenger RNA (mRNA) and TNF-‍α protein expression determination by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. Adiponectin and adiponectin receptor (adipoR) expression was measured by RT-PCR, quantitative real-time PCR (qPCR), immunohistochemical analysis, and cell counting kit-8 (CCK-8) cell proliferation experiments. We then quantified TNF-α and α-SMA mRNA and protein expression levels by qPCR and immunofluorescence (IF) staining. RNA interference (RNAi) was used to explore the function of the adipoR genes. To investigate the signaling pathway, we applied western blotting (WB) to examine phosphorylation of adenosine 5'-monophosphate (AMP)‍-activated protein kinase (AMPK). In vivo, an adiponectin (APN)‍-knockout (APN-KO) mouse model mimicking adventitial inflammation was generated to measure TNF-α and α‍-SMA expression by application of qPCR and IF, with the goal of gaining a comprehensive atlas of adiponectin in vascular remodeling. RESULTS: In both cells and tissues, UII promoted TNF-α protein and TNF-α-R secretion in a dose- and time-dependent manner via Rho/protein kinase C (PKC) pathway. We detected marked expression of adipoR1, T-cadherin, and calreticulin as well as a moderate presence of adipoR2 in AFs, while no adiponectin was observed. Globular adiponectin (gAd) fostered the growth of AFs, and acted in concert with UII to induce α-SMA and TNF-α through the adipoR1/T-cadherin/calreticulin/AMPK pathway. In AFs, gAd and UII synergistically induced AMPK phosphorylation. In the adventitial inflammation model, APN deficiency up-regulated the expression of α-SMA, UII receptor (UT), and UII while inhibiting TNF-‍α expression. CONCLUSIONS From the results of our study, we can speculate that UII induces TNF‍-‍α protein and TNF-‍α‍-R secretion in AFs and rat tunica adventitia of vessels via the Rho and PKC signal transduction pathways. Thus, it is plausible that adiponectin is a major player in adventitial progression and could serve as a novel therapeutic target for cardiovascular disease administration.
Mice ; Rats ; Animals ; Adventitia/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Calreticulin/metabolism* ; Vascular Remodeling ; AMP-Activated Protein Kinases/metabolism* ; Cells, Cultured ; RNA, Messenger/genetics* ; Inflammation

Objective:To investigate the value of high-frequency ultrasound and shear wave elastography in preoperative evaluation of basal cell carcinoma (BCC) .Methods:A total of 95 patients with histopathologically confirmed cutaneous BCC were enrolled from Department of Dermatology, Hospital of Traditional Chinese Medicine of Zhongshan from January 2017 to December 2020, all of whom had underwent preoperative conventional ultrasonography and shear wave elastography. Conventional ultrasonography parametres including the maximum diameter, maximum infiltration depth, maximum blood flow velocity and resistance index were recorded, so were shear wave elastography parametres including the average Young′s modulus (Eave) , Young′s modulus standard deviation (Esd) and average Young′s modulus ratio (Eratio) . All the patients were divided into high- and low-risk BCC groups according to pathologic subtypes. Paired t-test was used to compare conventional ultrasonography and shear wave elastography findings between the 2 groups. Results:There were 15 cases in the high-risk BCC group and 80 cases in the low-risk BCC group. Compared with the low-risk BCC group, the high-risk BCC group showed significantly increased maximum depth of tumor infiltration (8.5 ± 4.6 mm vs. 4.5 ± 1.6 mm, t = 6.150, P < 0.001) , Eave (32.7 ± 11.2 kPa vs. 20.6 ± 5.1 kPa, t = 4.065, P = 0.001) and Esd (7.0 ± 4.1 kPa vs. 4.2 ± 2.1 kPa, t = 2.632, P = 0.018) , while there were no significant differences in the other measurement data between the two groups (all P > 0.05) . The areas under the receiver operating characteristic curves of the maximum infiltration depth, Eave and Esd for the diagnosis of high-risk BCC were 0.775, 0.909 and 0.822 respectively, and Eave showed the best diagnostic performance. Using 25.7 kPa as the cut-off value, the sensitivity and specificity of Eave were 86.7% and 85.0% for the diagnosis of high-risk BCC, respectively. Conclusion:High-frequency ultrasound and shear wave elastography can facilitate differential diagnosis between high- and low-risk BCC.

OBJECTIVE: To explore the genetic basis for a Chinese patient featuring cleidocranial dysplasia(CCD). METHODS: Genomic DNA was extracted from peripheral blood samples of the patient and his parents. Whole exome sequencing (WES) was carried out for the patient, and suspected variant was verified by Sanger sequencing. RESULTS: WES has identified a missense c.460G>T (p.Val154Phe) (GRCh37/hg19) variant of the RUNX2 gene. The variant was located in the Runt domain, a highly conserved region (PM1); it was not present in either the Genome Aggregation Database or the 1000 Genomes Project (PM2), and was predicted to have a deleterious effect on the gene product by multiple in silico prediction tools (PP3); the clinical phenotype of the patient was highly consistent with that of cleidocranial dysplasia (PP4). Furthermore, the variant was unreported in medical literature and was absent in both parents (PS2). Based on the American College of Medical Genetics and Genomics guidelines, the c.460 G>T variant of RUNX2 gene was predicted to be pathogenic (PS2+PM1+PM2+PP3+PP4). CONCLUSION The c.460G>T (p.Val154Phe) variant of the RUNX2 gene probably underlay the clinical phenotype in the patient. Above finding has enabled accurate diagnosis and expanded the spectrum of RUNX2 variants.
China ; Cleidocranial Dysplasia/genetics* ; Core Binding Factor Alpha 1 Subunit/genetics* ; Humans ; Mutation ; Whole Exome Sequencing

Leptin, a peptide hormone discovered in the 1990s, arouses interest as it can regulate the body′s metabolism. In recent years, many studies have shown that leptin can promote the proliferation, activation and cytokine synthesis of various immune cells, and participate in innate and adaptive immune responses. This article reviewed the role of leptin and involved signaling pathways in immune response and the potential of leptin as a vaccine adjuvant.

Here we investigate the physical and chemical properties of chiral self-assembling peptides and the role of uterine trauma regeneration. The circular dichroism was used to analyze secondary structure of chiral self-assembled peptide, and Congo red staining was used to observe the macroscopic process of peptide self-assembling. Erythrocyte lysis assay was used to examine the cleavage of peptide on cell membrane. The nanofiber scaffolds self-assembled by Chiral self-assembling peptides were used as the three-dimensional culture material to observe the growth effect of Hela cell. CCK-8 (cell counting kit-8) was used to study cell viability level between 2D (2-dimensional) and 3D (3-dimensional) culture environment. Rats endometrium curettage model was founded to evaluate the changes by immunohistochemistry staining and and HE staining. The secondary structure of chiral self-assembling peptides was stable β-sheet, and peptide could form dense membrane structure after 24 hours self-assembling cultured in salt ions. There was no harmful for the cell membrane of the peptide before and after self-assembling. Animal experiments show that chiral self-assembling peptide can significantly reduce the inflammatory response, promote the production of neovascularization, and accelerate the repair process. Chiral self-assembling peptide, as a new type of scaffold material, can construct a three-dimensional cell culture environment and used to repair uterine trauma.
Animals ; Endometrium ; Female ; HeLa Cells ; Humans ; Nanofibers ; Peptides ; Rats ; Regeneration

Objective? To explore the effect of quality control circle activities on improving the correct rate of nursing orders for critically ill patients. Methods? A total of 21 senior responsible nurses in ICU, Cardiology Department (CCU), Cerebrovascular Disease Specialty and Respiratory Department of the Sixth Affiliated Hospital of Guangzhou Medical University were selected as the research objects. Meanwhile, 434 critically ill patients admitted in the above four departments in April 2018 were selected as the pre-activity group of quality control circle. The 419 critically ill patients admitted in October 2018 were compared and analyzed in the post-qcc group. The correct rate of nursing orders of critically ill patients, the qualified rate of basic nursing, posture nursing, the incidence of pressure sores, falls or falling out of bed, and incontinence related dermatitis, the mastery rate of senior responsible nurses on assessment knowledge and the time required for the first nursing order to be issued, the mastery rate of disease condition of critical patients by responsible nurses, and the comprehensive literacy scores of members of quality control circle team before and after quality control circle activities were compared. Results? After QCC activities, the correct rate of nursing orders for critical patients, the qualified rate of basic nursing and postural nursing, the mastery rate of senior responsible nurses' knowledge of assessment and the mastery rate of nurses' conditions for critical patients were significantly higher than those before QCC activities, and the time needed for the first nurses' orders was less than that before QCC activities, with statistical difference (P＜ 0.01). The incidence of pressure ulcers, falls or falling out of bed, and incontinence related dermatitis was lower than that before the activity, with no significantly statistical difference (P＞ 0.05). After the activity, the nurses' abilities of quality control, team cohesion, sense of responsibility, coordination and communication, self-confidence and quality improvement were all improved, with statistical difference (P＜ 0.01). Conclusions? The management based on the quality control circle can effectively improve the correct rate of nursing orders for critically ill patients, improve the nursing quality of critically ill patients, the working efficiency and overall literacy of nurses.

Objective To prepare dual-targeted pH-sensitive DOX prodrug-microbubble complex and explore the characterization of complex with ultrasound as well as drug release in vitro . Methods Dual-targeted ligands ,cRGD and folate were conjugated with heparin using carbodiimide method ,and then the dual-targeted pH-sensitive DOX prodrug was synthesized by coupling DOX via a pH-sensitive hydrazone bond . The prodrug was combined with microbubbles to prepare complex by biotin-avidin system . The characterization of complex with/without ultrasound was investigated for size ,morphology and drug loaded capacity .In vitro drug release manner of complex with/without at different pH was analyzed . Results DOX content of the prodrug determined by UV Spectrophotometry was about 18 .9％ . Dynamic laser light scattering analysis( DLS) ,corresponding to transmission electron microscope( TEM ) findings ,revealed its inhomogeneous size distribution [ mean size ( 159 .7 ± 24 .5) nm and ( 1089 .0 ± 174 .9) nm ] . However ,the complex was dispersed into uniform fragment after ultrasound irradiation [ mean size ( 155 .9 ± 29 .8) nm , polymer dispersity index( PDI) 0 .22 ,Zeta potential - ( 20 .6 ± 3 .4) mV ] . The cumulative release rate of DOX from both complex and complex with ultrasound at pH 5 .0 were much faster than that at pH 7 .4 , displaying a pH-triggered release manner . Conclusions Dual-targeted pH-sensitive DOX prodrug-microbubble complex displays excellent drug release activity in acid environment . Uniform fragment and smaller particle size of complex could be achieved via ultrasound irradiation ,promoting DOX accumulation within tumor tissue and facilitating in vivo antitumor ability .

Objective To explore the diagnostic and score value of ultrasound on hemophiliac arthropathy referring to MRI on the diagnosis and score of hemophiliac arthropathy Methods The ultrasound and MRI examinations were performed on 42 joints of 42 hemophilia patients 14 knees 14 ankles and 14 elbows The consistency of ultrasound and magnetic resonance imaging in the detection and score of joint diseases was compared Finally inter-and intra-observer agreement of ultrasound scoring system were tested Results The consistency of ultrasound and magnetic resonance imaging was excellent κ=0 763-0 896 P < 0 001 in the detection of early soft tissue lesions effusion or hemarthrosis synovial hypertrophy hemosiderin excellent κ=0 793 P <0 001 in the detection of cartilage loss poor κ=0 133 P = 0 132 in the detection of erosions and poor κ= 0 100 P = 0 137 in the detection of subchondral cysts The consistency of ultrasound and magnetic resonance imaging was good to excellentκ=0 684-0 833 P < 0 001 in the score of early soft tissue lesions effusion or hemarthrosis synovial hypertrophy and hemosiderin and poor to good κ=0 145 -0 635 P <0 001 in the score of advanced osteochondral lesions cartilage loss and bone erosions The inter-observer agreement was good to excellent κ=0 676-0 870 P <0 001 for early soft tissue lesions and moderate to excellent κ=0 421- 0 75 1 P < 0 001 for advanced osteochondral lesions The intra-observer agreement was good to excellent κ=0 705-0 885 P <0 001 for early soft tissue lesions and moderate to good κ=0 532 -0 732 P <0 001 for advanced osteochondral lesions Conclusions Ultrasound plays an important role in detecting early soft tissue changes effusion or hemarthrosis synovial hypertrophy hemosiderin and cartilage loss which helps follow-up and guide clinical treatment.