Objective To evaluate the application of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) technology in the identification of filamentous fungi,and analyze the susceptibility of filamentous fungi to commonly used antibiotics.Methods A total of 100 strains of filamentous fungi were collected and identified rapidly by MALDI-TOF MS.The obtained results were compared with those from microscopic examination.The susceptibility of filamentous fungi was detected by the Etest method.Results Among 100 strains of filamentous fungi identified by MALDI-TOF MS,61 reached to the species level(score≥2.000),36 to the genus level(score between 1.700 and 1.999),and 3 failed to be identified (score ＜ 1.700).There was inconsistent results for one strain of filamentous fungi between MALDI-TOF MS and microscopic examination.The MIC90 of amphotericin B against Epidermophytonfloccosum was 0.19 μg/mL,while that against Aspergillus flavus was above 32 μg/mL.The MIC90 of itraconazole against Trichophyton tonsurans,Microsporum canis and Epidermophytonfloccosum were all below 0.38 μg/mL,while that against Aspergillus niger was above 32 μg/mL.The MIC90 of fluconazol were above 256 μg/mL for most of strains.The MIC90 of voriconazole and caspofungin against Aspergillus fumigatus,Aspergillus flavus,Aspergillus niger,Trichophyton rubrum,Trichophyton tonsurans and Microsporum canis were ≤0.38 μg/mL and ≤ 1 μg/mL,respectively.Conclusion The MALDI-TOF MS technology may be used to identify the filamentous fungi isolated from clinical specimens quickly,accurately and high-throughput.Voriconazole and caspofungin have effective anti-filamentous fungi activity.

n be used as a potential marker in the diagnosis of PCa. However, it can not be used as an index to monitor tumor progression or prognosis in PCa patients.

Objective To explore hypermethylation of RARβ2, GSTP1 and DAPK gene in prostate cancer tissues, and to explore its correlation with clinicopathological features of prostate cancer and its diagnostic value. Methods Hypermethylation of RARe2, GSTP1 and DAPK gene was detected by nested methylation-specific PCR (NMSP) in 57 prostate cancer (PCa) tissues and 35 benign prostate hyperplasia (BPH) tissues. The correlation between hypermethylation and clinicopathological features of prostate cancer and its diagnostic value were analyzed. Results The hypermethylation frequencies of RARβ2, GSTP1 and DAPK gene in PCa were significantly higher than those in BPH (RARβ2: 52.6% vs. 0% GSTP1: 61.4% vs. 2.9%;DAPK: 43.9% vs. 8.6%;all P<0.01). The methylation rate of RARβ2 gene was directly correlated with distinct Gleason scores and clinical stage (4～7 score vs. 8～10 score: 34.8% vs. 64.7%;stage B, C vs. stage D: 37.0% vs. 66.7%;x2=4.927 and 5.004, P=0.026 and 0.025). The methylation rate of GSTP1 gene was significantly different in patients with different Gleason scores (4～7 score vs. 8～10 score: 43.5% vs. 73.5%;x2 =11.530, P=0.001), but had no difference in patients with distinct clinical stage (stage B, C vs. stage D: 51.9% vs. 70.0%;x2=1.975, P=0.16) . There was no difference in DAPK gene methylation rate among patients with different Gleason scores and distinct clinical stage (4 ～7 score vs. 8～10 score: 39.1% vs. 50.0%;stage B, C vs. stage D: 33.3% vs. 53.3%;x2= 1.290 and 2.309, both P～0.05). GSTP1 gene showed the highest sensitivity of 61.4% (35/57)with specificity of 97.1%(34/35), while RARβ2 gene had the highest specificity of 100% (35/35) with the sensitivity of 52.6% (30/57). The sensitivity and specificity of DAPK gene were 43.9% and 91.4% (25/57 and 32/35), respeetively. When the hypermethylation of RARβ2, GSTP1 and DAPK gene were detected together, the diagnostic sensitivity was increased, but the specificity was decreased. Conclusions The aberrant methylation of RARβ2, GSTP1 and DAPK gene is correlated with tumorigenesis and progression of prostate cancer, which may be used as an effective diagnostic marker for prostate cancer.

Objective:In this study,a series of experiments were conducted to research the mechanism of anticancer and preliminary molecular effects of PAMs on the HEPG-2 cancer cells.Methods:Morphological observation and MTT assay were used to explore the inhibition and killing effect of PAMs acting on HEPG-2.AO/EB staining and Annexin V-FITC/PI staining were employed to observe the apoptosis of HEPG-2 treated with PAMs.The expression level of Foxm1,bcl-2 and others genes in HEPG-2 cells were detected by using qRT-PCR and western blot.Wound healing and transwell experiments determined if PAMs can inhibit the migration of HEPG-2.Results:PAMs can inhibit and kill HEPG-2 cells in time and dose-dependent manners,and the cytotoxic effects were closely related to the cell apoptosis.The mRNA expression of foxm1,bcl-2 and surviving gene were remarkably decreased in HEPG-2 cells after the treatment of PAMs.PAMs decreased the FoXM1 protein expression in HEPG-2 cells,while up-regulating thep53 protein expression.,and it could also inhibit the migration of cancer cells.Conclusions:The possible molecular mechanism for the killing of HEPG-2 cancer cells by PAMs was proposed.By down-regulating the expression of foxm1 and up-regulating the expression of p53,the transcriptional expression of their downstream target genes survivin and bcl-2 was inhibited or reduced,hence enhancing the cancer cell apoptosis.This study provides an important foundation for the development of anti-cancer Chinese folk medicine based on PAMs.

Objective: To evaluate the feasibility and efficacy of naked-eye assessment (NA) of ultrasound-guided fine needle aspiration cytology(US-FNAC) smears, which was performed by a trained non-cytological physician. Methods: A total of 290 smears of FNAC in 143 thyroid nodules were used to evaluate the value of NA by an assistant with more than two years experience of intervention with ultrasound guidance. NA results such as the background of smear (bloody/non-bloody), thickness (thick/thin), as well as the contents (granulated/non-granulated) were recorded. The correlation between NA and cytological results was analyzed. Number of cells under microscopy, the non-diagnostic rate, and the significance between benignity and malignancy with different features of specimens were compared. Results: There was no significant difference between the NA background and cytological findings(P=0.707, 0.200, 0.705, respectively). There was significance in number of cells, non-dianostic rate between thick and thinner specimens (P=0.000, 0.001, respectively). However, there was no significance in benignity and malignancy between thick and thinner specimens (P=0.385). There was significance in cytology results between the granulated and non-granulated smears(all P=0.000). The difference was found in 51.1% of the smears with granulates were malignancy in cytology. While 77.8% of the non-granulated smears were ultimately diagnosed as benignity. Conclusions The NA could be performed by a trained non-cytological physician and contribute to preliminary predictions of cytology effect.

In this study, the leukemia K562 cell line was used as a model to elucidate the anticancer effects and preliminary mechanisms of PAMs. MTT assay showed that PAMs could cause cytotoxicities in K562 cells in dose- and time-dependent manners. AO-EB, Annexin-FITC/PI staining showed that the killing effects of PAMs in K562 cells were related to apoptosis, which was further confirmed by the following molecular and enzymatic assay. The mRNA levels of pro-apoptotic genes caspase-3, caspase-9 and bax were remarkably increased while the anti-apoptotic gene bcl-2 was significantly decreased determined by fluorescent quantitative PCR. Western blotting disclosed that PAMs could up-regulate caspase-3 and down-regulate anti-apoptotic survivin protein expression. The latter was also consistent with the results that PAMs could increase the enzymatic activities of both caspase-3 and caspase-9. All these results suggested that PAMs could effectively inhibit the proliferation of K562 cells and the mechanisms may be closely related to apoptosis induction. The work provides evidence basis for PAMs to be potentially developed as anti-cancer leukemia Chinese medicine.

Objective:To analyze the characteristics of lung function in patients with spinal muscular atrophy (SMA) to provide evidence for multidisciplinary management of SMA.Methods:A total of 30 patients with SMA treated in the SMA multidisciplinary clinic of the Children′s Hospital, Zhejiang University School of Medicine from July 2019 to March 2021 were enrolled, including 1 child with type I, 18 patients with type Ⅱ and 11 children with type Ⅲ.There were 17 males and 13 females; the age ranged from 4 years to 21 years and 10 months old.A retrospective study was conducted to analyze the clinical features, spinal imaging findings and lung functions of patients with different clinical types of SMA and explore the factors influencing the lung functions of patients with SMA.Pulmonary function was measured by forced expiratory flow-volume curve.Forced vital capacity (FVC), forced expiratory volume in one second (FEV 1), FEV 1/FVC and peak expiratory flow (PEF) were measured.The results were expressed as the percentage of the measured value to predicted value.The Cobb angle was measured to evaluate scoliosis. Pearson correlation analysis and multiple linear regression analysis were used to evaluate the relationship between lung function and age and Cobb angle in patients with type Ⅱ SMA. Pearson correlation analysis and univariate linear regression analysis were used to evaluate the relationship between Cobb angle and age in patients with type Ⅱ SMA. Results:Pulmonary function in 1 type I patient showed decreased FVC and FEV 1; Among 18 patients with type Ⅱ, 14 cases had abnormal lung function (77.8%): FVC decreased in 12 patients (66.7%), FEV 1 decreased in 10 patients (55.6%), PEF decreased in 12 patients (66.7%). Among 11 patients with type Ⅲ, one had decreased FVC (9.1%). FVC, FEV 1 and PEF of patients with type Ⅱ were significantly lower than those of patients with type Ⅲ [(62.4±31.8)% vs.(90.8±11.0)%, (66.3±33.3)% vs.(97.8±9.9)%, (65.3±30.1)% vs.(98.6±21.1)%, all P<0.01]. Pearson correlation analysis showed that FVC of patients with type Ⅱ SMA was correlated with age and Cobb angle ( r=-0.864, -0.865, all P<0.001), FEV 1 was correlated with age and Cobb angle ( r=-0.878, -0.863, all P<0.001), PEF was correlated with age and Cobb angle ( r=-0.831, -0.783, all P<0.001), and Cobb angle was related to age ( r=0.922, P<0.001). Multiple linear regression analysis indicated that FVC of patients with type Ⅱ SMA was linearly correlated with Cobb angle ( R2=0.748, P<0.001), FEV 1 was linearly correlated with age ( R2=0.770, P<0.001), PEF was linearly related to age ( R2=0.690, P<0.001). Univariate linear regression analysis revealed that Cobb angle of patients with type Ⅱ SMA was linearly related to age ( R2=0.851, P<0.001). Conclusions:FVC, FEV 1 and PEF may decrease in patients with SMA.The degree of lung function damage is different in different types of SMA patients.With the increase of age, Cobb angle increases and FVC, FEV 1 and PEF decrease in patients with type Ⅱ SMA.Understanding the factors influencing the pulmonary function damage in patients with SMA is conductive to carrying out individual multidisciplinary management.

OBJECTIVE To establish the method for monitoring the dynamic changes of odor components in Cornus officinalis during processing . METHODS The decoction pieces of C. officinalis with different processing time were prepared by the wine steaming method . The dynamic changes of odor components were obtained by using ultra -fast gas electronic nose ；odor components were identified by comparing with AroChemBase database ；the dynamic changes of odor compounds were analyzed in combination with peak area ，and the chemical pattern recognition analysis were carried out . RESULTS A total of 12 common peaks of odor components were identified in the fingerprints of raw C. officinalis，and 21 in the fingerprints of decoction pieces of C. officinalis. Eight odor components with the high proportion of peak area during processing were ethanol ， isopropyl alcohol ， 2- methylpropylaldehyde，ethyl acetate ，2-methylbutanal，isoamyl alcohol ，2-hexanol and furfural ，among which ，the peak areas of ethanol，isoamyl alcohol and 2-hexanol showed a trend of first increasing and then decreasing ；at 24 h of processing ，their peak areas were still higher than those of raw products . The peak areas of ethyl acetate ，2-methylbutanal and furfural nearly increased with the increase of processing time . Variable importance in projection of above eight odor components were all greater than 1. CONCLUSIONS The method is established for monitoring the dynamic changes of odor components of C. officinalis during processing. Eight odor components such as ethanol can be used as monitoring indicators of C. officinalis dring processing .

OBJECTIVES: To investigate the clinical efficacy and safety of salbutamol in the treatment of children with later-onset spinal muscular atrophy (SMA). METHODS: This study is a prospective single-arm phase Ⅲ clinical study. Pediatric patients with SMA type Ⅱ and Ⅲ who visited Department of Neurology, Children's Hospital, Zhejiang University School of Medicine from December 2020 to June 2022 were enrolled. All patients were evaluated with motor function scales, pulmonary function test and drug safety before study. Patients were treated with salbutamol tablets orally, with an initial dose of 1 mg (tid). If tolerable, the dose was increased to 1.5 mg (tid) in the second week, then increased to 2 mg (tid) from the third week and maintained for 6 months. Patients were followed up at 1, 3 and 6 months of treatment. RESULTS: Twenty-six patients were enrolled, including 10 boys and 16 girls. There were 16 cases of SMA type Ⅱ and 10 cases of type Ⅲ with age at treatment initiation of 5.67 (3.13, 7.02) years and disease duration of 2.54 (1.31, 4.71) years. The Hammersmith Functional Motor Scale-Expanded (HFMSE) scores were increased from 14.0 (6.5, 43.0) before treatment to 26.0 (15.0, 46.5) after treatment (Z=－4.144, P<0.01) in 25 cases. The Revised Upper Limb Module Scale scores were increased from 33.0 (25.5, 36.0) before treatment to 35.0 (31.0, 36.5) after treatment (Z=－2.214, P<0.05) in 9 cases. In 7 ambulant children with SMA type Ⅲ, the six minutes walking distance was increased by 30 (15, 52) m after a 6-month treatment (Z=－2.366, P<0.05). Compared with the baseline pulmonary functions the patients showed a significant increase in forced vital capacity (FVC), forced expiratory volume in one second (FEV₁), and peak expiratory flow (PEF) in 15 cases after treatment (all P<0.05). According to patients and caregivers subjective reporting, there were various degrees of improvement in coughing, sputum production ability and exercise endurance. No serious adverse events were observed during the study. CONCLUSIONS Short-term oral administration of salbutamol may improve motor and pulmonary functions in later-onset SMA children with good safety.
Male ; Female ; Humans ; Child ; Albuterol/therapeutic use* ; Prospective Studies ; Muscular Atrophy, Spinal/drug therapy* ; Spinal Muscular Atrophies of Childhood/drug therapy* ; Treatment Outcome