Retrospective analysis of 201 clinical isolates of Aspergillus fumigatus from a hospital in Nanjing: clinical characteristics of infected patients and azole resistance / 中华皮肤科杂志

Objective:To summarize clinical characteristics of patients with Aspergillus fumigatus infection in a hospital in Nanjing, to preliminarily assess azole resistance in clinical isolates of Aspergillus fumigatus, and to investigate risk factors for the emergence of azole-resistant Aspergillus fumigatus. Methods:Clinical isolates of Aspergillus fumigatus were collected from inpatients in Department of Laboratory, Nanjing Drum Tower Hospital from March 2017 to February 2021. Clinical data on these infected patients were analyzed, azole sensitivity testing and mutation analysis of the cyp51A gene and its promoter region were performed for these Aspergillus fumigatus isolates. Results:A total of 201 strains of Aspergillus fumigatus were collected, and mainly isolated from sputum specimens. Among the infected patients, there were 131 males and 70 females, and their age were 64.2 ± 15.8 years. The patients were mainly collected from department of respiratory medicine (79 cases), department of intensive medicine (34 cases), department of rheumatology (19 cases), etc. Among these patients, common underlying diseases included interstitial pneumonia (32 cases), malignant tumors (18 cases), pneumonia (13 cases), trauma (12 cases), systemic lupus erythematosus (8 cases), etc. Drug susceptibility testing showed that 6 (2.99%) strains of Aspergillus fumigatus were resistant to itraconazole and posaconazole, and 3 patients infected with azole-resistant Aspergillus fumigatus had used antifungal drugs before testing. Sequencing was performed on the cyp51A gene and its promoter region in the 6 strains of azole-resistant Aspergillus fumigatus, and showed TR34/L98H/S297T/F495I mutation in 5 strains and TR34/L98H mutation in 1 strain. Conclusion:Compared with previously published data about azole resistance in China during 2010 -2015, the resistance of Aspergillus fumigatus to azoles in Nanjing Drum Tower Hospital did not increase from 2017 to 2021, and the mechanism of azole resistance was mostly associated with TR34/L98H/S297T/F495I mutation in the cyp51A gene and its promoter region.

Effects of salidroside on the secretion of inflammatory mediators induced by lipopolysaccharide in the co-culture of rat alveolar macrophages and type II alveolar epithelial cells / 生理学报

The aim of the present study was to investigate the effect of salidroside (Sal) on inflammatory activation induced by lipopolysaccharide (LPS) in the co-culture of rat alveolar macrophages (AM) NR 8383 and type II alveolar epithelial cells (AEC II) RLE-6TN. CCK-8 colorimetric method was used to detect cell proliferation percentage. The enzyme-linked immunosorbent assay (ELISA) was used to determine the content of tumor necrosis factor alpha (TNF-α), macrophage inflammatory protein-2 (MIP-2) and interleukin-10 (IL-10) in the supernatant. Western blot was used to examine the expression levels of phosphorylated AKT (p-AKT) and total AKT protein. The results showed that pretreatment of RLE-6TN cells or co-culture of RLE-6TN and NR 8383 cells with 32 and 128 µg/mL Sal for 1 h, followed by continuous culture for 24 h, significantly increased the cell proliferation (P < 0.05). Compared with control group, 32 and 128 µg/mL Sal pretreatment significantly increased the ratio of p-AKT/AKT in RLE-6TN cells (P < 0.05). Pretreatment of 32 µg/mL Sal not only inhibited the secretion of TNF-α and MIP-2 by NR 8383 cells induced by LPS (P < 0.05), but also enhanced the inhibitory effect of RLE-6TN and NR 8383 cells co-culture on the secretion of TNF-α and MIP-2 by NR 8383 cells induced by LPS (P < 0.05). In addition, 32 µg/mL Sal pretreatment promoted LPS-induced IL-10 secretion by NR 8383 cells (P < 0.05), and enhanced the promoting effect of co-culture of RLE-6TN and NR 8383 cells on the IL-10 secretion by LPS-induced NR 8383 cells (P < 0.05). In conclusion, Sal may directly inhibit LPS-induced inflammatory activation of AM (NR 8383), promote the proliferation of AEC II (RLE-6TN) through PI3K/AKT signaling pathway, and enhance the regulatory effect of AEC II on LPS-induced inflammatory activation of AM.

Protective effect of synthetic salidroside on acute lung injury in rats / 生理学报

To study the protective effect and mechanism of synthetic salidroside on acute lung injury (ALI) induced by lipopolysaccharide (LPS), male Sprague-Dawley (SD) rats were randomly divided into saline control group, 3 mg/kg LPS model group, different doses of salidroside groups (5, 20 and 80 mg/kg), and 5 mg/kg dexamethasone group. Intratracheal LPS instillation was used to establish the ALI model 0.5 h after intraperitoneal injection of salidroside or dexamethasone, and the rats were sacrificed 6 h later. Lung wet/dry weight ratio (W/D) was calculated. Lung tissue pathology and lung injury score (LIS) were observed and evaluated through hematoxylin and eosin (HE) staining. The centrifugal sediment of bronchoalveolar lavage fluid (BALF) was used to count the polymorphonuclear leukocyte (PMN) number by Wright's staining, and the centrifugal supernatant of BALF was used to determine the contents of protein and inflammatory factors (TNF-α, IL-1β and IL-6). The contents of myeloperoxidase (MPO) and malondialdehyde (MDA) in lung tissue were determined. Western blot was used to detect the expression levels of phosphorylated and total nuclear factor kappa B (NF-κB)/p65 protein in lung tissue. The results showed that, compared with LPS group, the intervention of synthetic salidroside alleviated the pathological damage in lung tissue, decreased the LIS and lung W/D ratio (P < 0.05), reduced the PMN number, the contents of protein and inflammatory factors in BALF (P < 0.05), reduced the contents of MPO and MDA in lung tissue (P < 0.05), and inhibited the expression of p-NF-κB in lung tissue (P < 0.05). The results suggest that synthetic salidroside has a protective effect on ALI induced by LPS, and its mechanism is related to inhibiting the phosphorylation of NF-κB and reducing the aggregation of PMN in the lung.

Mechanism of SARI gene promoting apoptosis in acute myeloid leukemia cells / 中国免疫学杂志

Objective:To explore the apoptotic mechanisms of acute myeloid leukemia(AML) cells by a lentivirus mediated-SARI(suppressor of AP-1,regulated by IFN) overexpression system.Methods: The lentiviral vectors of overexpression SARI were transfected into AML cell HL-60 and NB4.Real-time quantitative PCR and Western blot were employed in the following evaluation tests.Both the cells were all divided into blank control group(CON group),empty vector control group(NC group)and SARI overexpression group(SARI group).Western blot was used to detect the expressions of apoptosis-related proteins Bcl-2,Bcl-xl,Bax and apoptotic pathway related proteins CytoC,Caspase8,Caspase9,Caspase3,Actived-Caspase3,PARP.Results:The SARI groups exhibited a high level of SARI mRNA and SARI protein when compared with CON group and NC group(P<0.05).In both the SARI groups,the expression of anti-apoptotic proteins Bcl-2,Bcl-xl were down-regulated and the pro-apoptotic protein Bax was up-regulated.CytoC increased,Caspase8,Caspase9 and Caspase3 decreased,Actived-Caspase3 up-regulated.PARP down-regulated,and PARP spliced variant up-regulated.Conclusion: SARI might promote the apoptosis of AML cells through activing the exogenous and endogenous apoptotic pathways which crossed to exogenous.

A case of cutaneous Arthrinium phaeospermum infection after lower limb amputation / 中华皮肤科杂志

A 59?year?old female patient, who received bilateral lower limb amputation 39 years ago, presented with eczematoid changes in both lower limbs for over 20 years, and with chronic granuloma?like lesions complicated by verrucous hyperplasia for more than 10 years. There were large areas of infiltrative and proliferative lesions with exudation and peripheral erythema at the amputation sites in both knee joints. The lesions were hard with tenderness on palpation. Microscopic examination of lesional scales with 10%KOH showed negative results for fungi. However, three times of culture on the Sabouraud dextrose agar(SDA)medium all grew the same kind of fungus, and the front side and reverse side of its filamentous colony were white and orange yellow respectively. Microculture showed that linear hyaline conidiophores came out from lageniform mother cells with conidia ascending alongside. The conidia looked like dark brown eye lens, with an equatorial germ slit. Based on these findings, this fungus was identified as Arthrinium phaeospermum. Periodic acid?Schiff (PAS) staining showed scattered hyphae in the stratum corneum. The internal transcribe spacer(ITS)sequence of the isolated fungus showed 99%consistency with that of Arthrinium phaeospermum. The patient was diagnosed with cutaneous Arthrinium phaeospermum infection, and treated with oral itraconazole capsules 200 mg/d for 16 days. One month later, follow?up showed satisfactory outcomes.

Clinical analysis on chronically recurrent and generalized superficial mycosis caused by trichophyton rubrum / 医学研究生学报

Objective There have been a few reports on chronically recurrent and generalized superficial mycosis caused by trichophyton rubrum.This article was to investigate the cause, diagnosis and therapy of the mycosis. Methods 5 patients with chron-ically recurrent and generalized superficial mycosis caused by trichophyton rubrum were collected from June 2012 to June 2014 in our hospital.Bacterioscopic examination and cultivation were made on skin lesions of the patients.A typical patient who had 7-year course of desease with toenails seriously infected and widespread skin eruption was selected for histopathology examination on skin lesions, mi-crobiology and molecular biology study on 4 bacterial strains isolated from skin lesions in different parts, and in vitro chemosensitivity assay for drug selection.PCR (rDNA ITS sequence analysis) was performed for diagnosis and early treatment. Results Microscopic examintion on skin lesions demonstrated numerous septate, branched hyphae.Cultivation and molecular biology study identified tricho-phyton rubrum.The strain was identified as trichophyton rubrum by ITS sequence analysis and the isolated strains from different lesions were the same fungal species.Histopathology examination revealed slight hyperplasia of squamous epithelium , epidermal hyperkeratini-zation and the upper dermis presented a sparse perivascular lymphocytic infiltrate.The PAS-stain confirmed the presence of few hyphae in the horny layer.The pathogen of this case was trichophyton rubrum. A combination therapy with systemic itraconazole and topically applied terbinafine hydrochloride cream was successful.A follow-up examina-tion one year later showed no recurrence of symptoms. Conclusion The isolation and identification of pathogen is the key to the diagnosis of chronically recurrent and generalized superficial mycosis, with ad-ditional attention to all or none toenail infection.The therapy should not focus simply on the tinea corporis, while comprehensive treatment combined with chemosensitivity assay is preferred.

Does diffusion tensor tractography of the corticospinal tract correctly reflect motor function?

To investigate the consistency of diffusion tensor tractography of the corticospinal tract on motor function. Clinical Presentations and Intervention: Three patients with brain tumor were admitted to our hospital with impaired motor function. Diffusion tensor imaging [DTI] and tractography were performed in these patients to assess their affected corticospinal tract. The corticospinal tract showed interruption with moderately impaired motor function in 2 patients. The third case had significantly weakened muscle strength on the left upper limb but an intact right corticospinal tract. These cases showed that the corticospinal tracts obtained by DTI with tractography were inconsistent with motor function. Hence, DTI should be interpreted with caution

HIV-1 infection up-regulating expression of interferon-stimulated gene 15 in cell lines / 病毒学报

To investigate whether HIV-1 infection affects expression of interferon-stimulated gene 15 (ISG15) and determine the antiviral effect of ISG15 in vitro, ISG15 expression at transcription and protein level and supernatant p24 of HIV-1 was detected in various HIV-1 infected or transfected cell lines, respec tively. HIV-1 molecular clone pNL4-3 was used to transfect 293T, TZM-bl and HeLa cells while HIV-1 pseudo-typed virus was used to infect Jurkat, MT-1 and THP-1 cells. After twenty-four hours post infection or transfection, cells were harvested for extraction of total RNAs and subsequently used in real time PCR for quantification of ISG15 transcriptional expression. After forty-eight hours post infection or transfection, cells were harvested for extraction of total proteins to detect ISG15 protein expression. A significant up-regulation of ISG15 at transcription level was observed in HIV-1 infected or transfected cell lines, particulaly in THP-1 and TZM-bl cells. Up-regulation of ISG15 protein was observed only in TZM-bl cell. Cotransfection of ISG15 and HIV-1 indicated that ISG15 inhibited production of HIV-1 progeny virus in a dose and time depend manner in 293T cell but not TZM-bl cell. These results revealed upregulating ISG15 expression in transcriptional level and potential antagonistic mechanism against ISG15 by HIV-1 infection, simultanelusly.

Comparison of mannitol and hypertonic saline in treatment of intracranial hypertension of rabbits / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To compare the effects of mannitol and hypertonic saline (HS) in treatment of intracranial hypertension (ICH) of rabbits.</p><p><b>METHODS</b>The animal mode of ICH was established by perfusing artificial cerebrospinal fluids (aCSF) with controlled pressure into the cerebral ventricles of rabbits. The mean arterial pressure, respiratory rate, tidal volume, perfusion rate of aCSF and water content of cerebrum were investigated in rabbits with ICH after a single bolus of 20% mannitol (5 ml/kg), 7.5% HS (2.2 ml/kg) or 23.4% HS (2.2 ml/kg).</p><p><b>RESULTS</b>After the intracranial pressure was elevated from 15 cmH₂O to 75 cmH₂O, the mean arterial pressure was increased and the tidal volume was decreased. After treatment by 20% mannitol, 7.5% HS or 23.4% HS, the increased percentage of mean arterial pressure and the decreased percentage of tidal volume were similar to the changes in control group. However, the perfusion rate of CSF was increased and water content of cerebrum was decreased after treatment by either 20% mannitol or 23.4% HS, but not by 7.5% HS. No different effects were found between 20% mannitol and 23.4% HS.</p><p><b>CONCLUSION</b>With the similar osmotic burden, 20% mannitol is more effective in treating ICH than 7.5% HS. With higher osmotic load, the efficacy of HS is enhanced, and 23.4% HS may be used as an alternative to mannitol in treatment of ICH.</p>

Effects of trichostatin A on the expressions of inflammatory cytokines and toll-like receptor 4 and the acetylation of nuclear factor-κB induced by lipopolysaccharide in macrophage / 生理学报

The present study aims to explore the possible mechanisms that trichostatin A (TSA), a histone deacetylases inhibitor (HDACi), affects the inflammatory signaling pathways of lipopolysaccharide/toll-like receptor 4/nuclear factor-κB (LPS/TLR4/NF-κB). Murine macrophage cell line RAW264.7 cells were employed. MTT assay was used to assess cell viability. The contents of TNF-α, IL-1β and IL-6 in culture supernatant were assayed by enzyme-linked immunosorbent assay (ELISA). TLR4 expression and NF-κB/p65 (Lys310) acetylation were examined by Western blotting. DNA binding activity of NF-κB/p65 was detected by using TransAM(TM) NF-κB/p65 activity assay kit. The results showed that, compared with control group, which was treated by DMSO, the cells treated with TSA (20, 40, 80 ng/mL) showed decreased percentages of cell survival (P < 0.05). The contents of TNF-α, IL-1β and IL-6 in culture supernatant were all increased by LPS (100 ng/mL), whereas reduced by 40 ng/mL TSA pretreatment (P < 0.05). TSA pretreatment inhibited LPS-induced up-regulation of TLR4 protein expression. Acetylation of NF-κB/p65(Lys310), which was already increased by LPS, was further enhanced by TSA (P < 0.05). On the contrary, LPS-increased DNA binding activity of NF-κB/p65 was decreased by pretreatment with TSA (P < 0.05). The results suggest that TSA-induced anti-inflammation may be attributed to decreases in the expression of TLR4 and DNA binding activity of NF-κB/p65.

Body vitamin D content and its relationship with body composition of children in Huairou district of Beijing / 中华流行病学杂志

Objective To analyze vitamin D concentration and its association with body composition of children in Huairou district of Beijing, to provide evidence for evaluation and improvement of nutritional status of vitamin D in children. Methods Totally, 381 children aged 7-11 years were recruited in Huairou district of Beijing(40.3°N). Samples of overnight fasting venous blood (drawn between 0630 and 0900) were obtained in late March. Serum 25-hydroxyvitamin D [25(OH)D] concentration was determined by ELISA kits (IDS Ltd, UK). Body composition indices of the whole body, the distal and proximal forearm were measured using dual-energy X-ray absorptiometry (DEXA, Norland, US A). Results The average serum 25 (OH) D concentration of all subjects was (44.4±12.5) nmol/L. The percentage of vitamin D insufficient [serum 25(OH) D≤ 50 nmol/L ] reached as high as 68.5% (261 people). The 25 (OH) D concentration of boys (46.3 nmol/L± 13.3 nmol/L) was significantly higher than that of girls (42.0 nmol/L±11.1 nmol/L), (t=3.38,P< 0.01). Between the serum 25(OH)D concentration and lean body mass of proximal forearm, distal forearm, whole body, and four limbs, significant positive correlations (r=0.13-0.19,P<0.05) were observed. After age, gender, height and weight were under controlled for, correlations between the serum 25(OH)D concentration and lean body mass at proximal forearm, and the two lower limbs still existed (r=0.12-0.14,P<0.05). The serum 25(OH)D concentration, the percentage of body fat at proximal forearm, and distal forearm were negatively correlated (r=-0.14, P<0.05;r=-0.11, P< 0.05). However, after adjusting for confounding effects, this correlation disappeared. No significant correlation between the serum 25 (OH)D concentration and body fat was observed at any position. Conclusion Vitamin D deficiency was common in children of suburb area of Beijing. Vitamin D status was positively associated with the lean body mass.