Three Chitin Deacetylase Family Members of Beauveria bassiana Modulate Asexual Reproduction and Virulence of Fungi by Mediating Chitin Metabolism and Affect Fungal Parasitism and Saprophytic Life.

As an important chitin-modifying enzyme, chitin deacetylase (CDA) has been characterized in many fungi, but its function in the entomopathogenic fungus Beauveria bassiana remains unclear. Three CDAs with conserved domains of the carbohydrate esterase 4 (CE-4) family were identified in B. bassiana. Disruption of CDA1 resulted in growth restriction of the fungus on medium with chitin as a carbon source or without a carbon source. Deletion of CDA1 and CDA2 led to defects in fungal conidial formation and conidial vitality compared with those of the wild type (WT), and the conidial yield decreased by 25.81% to 47.68%. Inactivation of three CDA genes resulted in a decrease of 20.23% to 27% in the blastospore yield. ΔCDA1 and ΔCDA3 showed 29.33% and 23.34% reductions in cuticular infection virulence, respectively. However, the CDA family may not contribute to hemocoel infection virulence. Additionally, the sporulation of the insect carcass showed that the three gene deletion mutants were 68.45%, 63.84%, and 56.65% less than WT. Penetration experiments with cicada wings and enzyme activity assays were used to further explore the effect of the fungus on chitin metabolism after gene deletion. Although the three gene deletion mutants penetrated the cicada wings successfully and continued to grow on the underlying medium, their colony sizes were reduced by 29.12% to 47.76%. The CDA enzyme activity of ΔCDA1 and ΔCDA3 decreased by 84.76% and 83.04%, respectively. These data showed that members of the CDA family play a different role in fungal growth, conidial quality, and virulence. IMPORTANCE In this study, we report the roles of CDA family in entomopathogenic fungus B. bassiana. Our results indicated that CDA modulates asexual development and regulates fungal virulence by altering chitin deacetylation and metabolic capacity. CDA affected the biological control potential and life history of B. bassiana by affecting its parasitic and saprophytic life. These findings provide novel insights into the roles of multiple CDA paralogues existing in fungal biocontrol agents.

Two new Clitocella species from North China revealed by phylogenetic analyses and morphological characters.

Two new species of Clitocella are proposed based on morphological and phylogenetic investigations. Clitocellaborealichinensis sp. nov. is closely related to C.orientalis but distinguished from the latter by its slightly smaller basidiospores and hyphae of pileipellis with pale brown to brown intracellular or parietal pigment. Clitocellacolorata sp. nov. is closely related to C.popinalis and C.mundula in macromorphology but is differentiated from C.popinalis by its slightly smaller basidiospores and the difference in genetic profile, and from C.mundula by its relatively colorful pileus (white to yellowish white, grayish white to grayish brown, pink white). Phylogenetic analyses based on sequence data from five different loci (ITS, nrLSU, tef1, rpb2 and atp6) support the taxonomic position of the two new species in the genus Clitocella. The illustrations and descriptions for the new taxa are provided.

The transcription factor Ron1 is required for chitin metabolism, asexual development and pathogenicity in Beauveria bassiana, an entomopathogenic fungus.

Ndt80-like transcription factor Ron1 is best known for its essential role in the regulation of N-acetylglucosamine (GlcNAc) catabolism. Ron1 was again found to be essential for sensing GlcNAc in Beauveria bassiana. Importantly, our study revealed that Ron1 is involved in the metabolic processes of chitin and asexual development. To further investigate the novel functions of Ron1 in B. bassiana, extracellular chitinase activity in the ΔRon1 mutant was found to decrease by 84.73% compared with wild type. The deletion of Ron1 made it difficult for the fungus to accumulate intracellular GlcNAc. Furthermore, transcriptomic analysis revealed that Ron1 exerted a significant effect on global transcription and positively regulated genes encoding chitin metabolism in respond to chitin nutrition. Yeast one-hybrid assay confirmed that Ron1 could bind to specific cis-acting elements in the promoters of chitinase and hexokinase. In addition, ΔRon1 displayed an impaired chitin component of the cell wall, with a chitin synthetase (ChsVII) predicted to function downstream of Ron1. Finally, the virulence of ΔRon1 mutant was significantly reduced in the Galleria mellonella insect model through cuticle infection or cuticle bypassing infection. These data functionally characterize Ron1 in B. bassiana and expand our understanding of how the transcription factor Ron1 works in pathogens.

New Species of Mallocybe and Pseudosperma from North China.

Within the family Inocybaceae, many species of Mallocybe and Pseudosperma have been reported, but there are only a few reports on these two genera from north China. In this study, six collections of Mallocybe and 11 collections of Pseudosperma were studied by morphological and phylogenetic methods. Phylogenetic analyses based on sequence data from three or two different loci (ITS, LSU, and rpb2 for Mallocybe; ITS and LSU for Pseudosperma) are performed to infer species relationships within genera Mallocybe and Pseudosperma, respectively. Results indicate that eight species of Mallocybe and Pseudosperma are found in Shanxi province, north China; two new species of Mallocybe, M. depressa and M. picea, are described. Overall, six species belong to Pseudosperma, of which three are new: P. gilvum, P. laricis and P. pseudoniveivelatum.

HEAD-US-C quantitative ultrasound assessment scale in evaluation of joint damage in patients with moderate or severe hemophilia A received on-demand versus prophylaxis replacement therapy / 中华血液学杂志

Objective: To explore the evaluation of joint injury by HEAD-US-C (Hemophilic Early Arthropathy Detection with UltraSound in China, HEAD-US-C) in patients with moderate or severe hemophilia A treated with prophylaxis vs on-demand. Methods: The patients from June 2015 to July 2017 with moderate or severe hemophilia A were examined by ultrasound imaging of the elbows, knees and ankles; Meanwhile the HEAD-US-C ultrasound assessment scale and hemophilia joint health score scale 2.1 (HJHS2.1) were used to score the joint status. The correlation between the HEAD-US-C and HJHS score was performed in prophylaxis group and on-demand group patients, respectively. Results: A total of 925 cases of joint ultrasonography were conducted in 70 patients with moderate or severe hemophilia A. Among patients with moderate hemophilia, the median (IQR) of HEAD-US-C score and HJHS score in on-demand group were significantly higher than those in the prophylaxis group[1 (0, 6) vs 0.5 (0, 3) , z=0.177, P=0.046],[2 (0, 4) vs 2 (0, 3) z=0.375, P=0.007], even though there was no significant difference of the median (IQR) number of annualized target joints bleeding episodes between on-demand and prophylaxis groups[1 (0, 7) vs 1 (0, 5) , z=1.271, P=0.137]. Unlike in moderate cases, on-demand treatment group had more annualized target joints bleeding episodes than prophylaxis group among patients with severe hemophilia[3 (0, 8) vs 2 (0, 8) , z=0.780 P=0.037]. The prophylaxis group compared favorably with on-demand therapy group in terms of HEAD-US-C score[1 (0, 6) vs 4 (0, 7) , z=2.189, P=0.008], and HJHS score[2 (0, 5) , 4 (1, 6) , z=3646, P<0.001]for the severe hemophilia patients. The positive correlation between HEAD-US-C score and HJHS score was identified (P<0.05) , whether on-demand treatment or prophylaxis groups. The correlation coefficient between HEAD-US-C score and HJHS score in on-demand treatment and prophylaxis groups were 0.739 (95% CI 0.708-0.708) , 0.865 (95% CI 0.848-0.848) respectively, and 95% CI didn't overlap (P<0.05) , indicating that the correlation coefficient in prophylaxis group had stronger correlation than that in on-demand group. Conclusions: Clinical effects of prophylaxis were significantly better than those of on-demand treatment in patients with moderate or se-vere haemophilia A. HEAD-US-C scoring system could effectively evaluate joints damage in hemophilia A patients treated with on-demand or prophylaxis, companied by significantly positive correlation with HJHS clinical evaluation system, and provided objective index for clinical effect assessment.

Discussion on controlling proportion of medical consumables / 医疗卫生装备

Objective To explore the method for controlling the proportion of medical consumables.Methods The objective and significance of controlling the proportion of medical consumables were introduced,and the method was analyzed from the aspects of bidding procurement,total control mechanism,administrative approval and involvement in department management by objectives.Results The effective management of medical consumables decreased irrationality and procurement price of medical consumables.Conclusion Controlling the proportion of medical consumables promotes public hospital reform and the economic burden of the patient.

Discussion on controlling proportion of medical consumables / 医疗卫生装备

Objective To explore the method for controlling the proportion of medical consumables.Methods The objective and significance of controlling the proportion of medical consumables were introduced,and the method was analyzed from the aspects of bidding procurement,total control mechanism,administrative approval and involvement in department management by objectives.Results The effective management of medical consumables decreased irrationality and procurement price of medical consumables.Conclusion Controlling the proportion of medical consumables promotes public hospital reform and the economic burden of the patient.

Contrast-enhanced ultrasound characteristics of papillary thyroid carcinoma and its relationship with microvessel density and microvessel area / 中华医学超声杂志（电子版）

Objective To explore the correlation between quantitative parameters of blood perfusion with contrast-enhanced ultrasound (CEUS) and microvessel density (MVD),microvessel area (MVA) in papillary thyroid carcinoma (PTC).And to investigate the value of CEUS in evaluating the angiogenesis in PTC before operation.Methods Totally 69 cases of patients with papillary thyroid carcinoma were selected from April 2014 to October 2016 in the Affiliated Hospital of Qingdao University.The CEUS characteristics of 69 patients with papillary thyroid carcinoma confirmed by pathology were retrospectively analyzed.The patients were divided into three groups according to maximum diameter of lesions (＜ 1 cm group,1-2 cm group and ＞ 2.0 cm group),and two groups according to pathologic reports (neck lymph node metastatic and nonmetastatic groups).The blood perfusion parameters between or among different groups were evaluated by ttest or one-way ANOVA.Immunohistochemical staining were performed to evaluate the MVD,MVA in the surgical specimens,and the correlation of quantitative parameters with MVD,MVA were assessed by Spearman.Results (1) Peak Intensity (Peak),area under the curve (AUC),MVD and MVA of thyroid carcinoma were lower than the surrounding normal thyroid tissue (14.95 ± 4.96 vs 22.67±6.11,970.01±263.20 vs 1798.35±563.67,118.91±31.32 vs 206.27±39.58,8.58±-2.68 vs 18.47±3.13),and the differences were statistically significant (t=-8.700,-11.061,-14.377 and-20.532,all P ＜ 0.05).(2)With the increase of the lesion's maximum diameter,Peak,AUC,MVD and MVA increased,and the differences were statistically significant (t=0.000,0.000,0.000,0.000;t=0.027,0.044,0.033,0.000;t=0.027,0.044,0.033,0.000,all P ＜ 0.05).(3) Papillary thyroid carcinoma with lymphatic involvement had significantly higher values of Peak,AUC,MVD and MVA than those without lymphatic involvement (16.86±4.36 vs 13.80±3.55,1128.16±290.85 vs 874.39±192.27,114.12±30.69 vs 103.67±22.19,10.30 ± 2.44 vs 7.54 ± 2.29),and the differences were statistically significant (t=3.177,4.366,6.336 and 4.742,all P ＜ 0.05).(4) A positive correlation existed between the Peak,AUC and MVD,and the differences were statistically significant (r=0.506,0.478,all P ＜0.05).Peak,AUC and MVA showed positive correlation,and the differences were statistically significant (r=0.648,0.653,all P ＜ 0.05).TP,MTT and MVD,MVA showed no correlations (all P ＞ 0.05).Conclusions The values of Peak and AUC calculated from CEUS were correlated to MVD and MVA.CEUS may be used to evaluated the angiogenesis of PTC before operation.And CEUS is helpful for prediction of prognosis of PTC.

Contrast-enhanced Ultrasound for Diagnosing Papillary Thyroid Carcinoma / 中国医学影像学杂志

Purpose To evaluate the diagnostic value of contrast-enhanced ultrasound (CEUS) in papillary thyroid carcinomas (PTC) by exploring the relationship between quantitative parameters of time-intensity curve (TIC) of CEUS for PTC and tumor size and metastasis of cervical lymph nodes.Materials and Methods 124 patients with PTC confirmed by surgery and pathology in the Affiliated Hospital of Qingdao University were retrospectively analyzed.According to maximum diameter of lesions (D),the lesions were divided into three groups with D＜1.0 cm,1.0 cm ≤ D ≤ 2.0 cm and D＞2.0 cm.The lesions were also divided into LN(+) group with lymph node metastasis and LN(-) group without lymph node metastasis based on pathology of cervical lymph nodes.The features of CEUS and quantitative parameters of TIC of the above groups were analyzed.Results ① The CEUS showed that the PTC nodules were mainly concentric and heterogeneous enhancement.Thyroid carcinoma with D＜1.0 cm and 1.0 cm≤D≤2.0 cm showed low enhancement (45/57,31/42),while thyroid carcinomas with D＞2.0 cm exhibited high enhancement (14/25),and the difference was significant (P＜0.05).① With the increase of the diameter of PTC,the peak intensity [(12.75 ± 3.77)％,(15.53 ± 3.62)％,(18.11 ± 4.28)％],the area under the curve [(820.52±289.19)％.s,(873.84± 156.19)％· s,(1118.8± 152.48)％ ·s] and the ratio of the perfusion defects (24.56％,52.38％,72.00％) were increased,and the differences were statistically significant (P＜0.05).③ The cervical lymph node metastasis rate of PTCs with isoenhancement or hyperenhancement patterns showed by CEUS was significantly higher than that with hypoenhancement (P＜0.05).The peak intensity and the area under the curve of LN (+) group were higher than that of LN (-) group,and the differences were statistically significant (P＜0.05).Conclusion There were significant differences in imaging features of CEUS between the PTC nodules with different size and lymph node metastasis,which can provide valuable information for clinical diagnosis.

Nickel-Refining Fumes Induced DNA Damage and Apoptosis of NIH/3T3 Cells via Oxidative Stress.

Although there have been numerous studies examining the toxicity and carcinogenicity of nickel compounds in humans and animals, its molecular mechanisms of action are not fully elucidated. In our research, NIH/3T3 cells were exposed to nickel-refining fumes at the concentrations of 0, 6.25, 12.50, 25, 50 and 100 µg/mL for 24 h. Cell viability, cell apoptosis, reactive oxygen species (ROS) level, lactate dehydrogenase (LDH) assay, the level of glutathione (GSH), activities of superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) level were detected. The exposure of NIH/3T3 cells to nickel-refining fumes significantly reduced cell viability and induced cell apoptotic death in a dose-dependent manner. Nickel-refining fumes significantly increased ROS levels and induced DNA damage. Nickel-refining fumes may induce the changes in the state of ROS, which may eventually initiate oxidative stress, DNA damage and apoptosis of NIH/3T3 cells.

Screening to Identify Commonly Used Chinese Herbs That Affect ERBB2 and ESR1 Gene Expression Using the Human Breast Cancer MCF-7 Cell Line.

Aim. Our aim the was to screen the commonly used Chinese herbs in order to detect changes in ERBB2 and ESR1 gene expression using MCF-7 cells. Methods. Using the MCF-7 human breast cancer cell line, cell cytotoxicity and proliferation were evaluated by MTT and trypan blue exclusion assays, respectively. A luciferase reporter assay was established by transient transfecting MCF-7 cells with plasmids containing either the ERBB2 or the ESR1 promoter region linked to the luciferase gene. Chinese herbal extracts were used to treat the cells at 24 h after transfection, followed by measurement of their luciferase activity. The screening results were verified by Western blotting to measure HER2 and ER α protein expression. Results. At concentrations that induced little cytotoxicity, thirteen single herbal extracts and five compound recipes were found to increase either ERBB2 or ESR1 luciferase activity. By Western blotting, Si-Wu-Tang, Kuan-Shin-Yin, and Suan-Tsao-Ren-Tang were found to increase either HER2 or ER α protein expression. In addition, Ligusticum chuanxiong was shown to have a great effect on ERBB2 gene expression and synergistically with estrogen to stimulate MCF-7 cell growth. Conclusion. Our results provide important information that should affect clinical treatment strategies among breast cancer patients who are receiving hormonal or targeted therapies.

Coprescription of Chinese herbal medicine and Western medication among female patients with breast cancer in Taiwan: analysis of national insurance claims.

BACKGROUND: Many female breast cancer (FBC) patients take Chinese herbal medicine (CHM) and Western medication (WM) concurrently in Taiwan. Despite the possibility of interactions between the CHM and WM mentioned in previous studies, the pattern of these coprescriptions in FBC patients remains unclear. Hence, the aim of the present study is to investigate the utilization of coprescriptions of CHM and WM among the FBC patients in Taiwan. METHODS: The study was a cross-sectional survey using the sampled cohort in 2009 obtained from the National Health Insurance Research Database in Taiwan. There were 3,507 FBC patients identified from the registry for catastrophic illness patients. Ambulatory visit records, corresponding prescriptions, and the data of beneficiaries belonging to the FBC patients were further extracted. A total of 1,086 FBC patients used CHM at least once. CHM and WM prescribed within any overlapping duration were defined as coprescriptions. RESULTS: There were 868 (80.0%) patients simultaneously receiving CHM and WM. A total of 4,927 CHM prescriptions and 6,358 WM prescriptions were prescribed concurrently. Among these coprescriptions, the most frequently used CHM was jia-wei-xiao-yao-san (21.2%), and the most frequently coprescribed WM was acetaminophen (38.9%), followed by tamoxifen (25.5%). There were 346 patients using systemic adjuvant therapy and CHM concurrently. The most commonly coprescribed CHM with chemotherapy, endocrine therapy, and trastuzumab was xiang-sha-liu-jun-zi-tang, jia-wei-xiao-yao-san, and zhi-gan-cao-tang, respectively. CONCLUSION: The combined use of CHM with WM is prevalent. The main purpose of combining CHM with systemic cancer treatment is to alleviate the treatment-related adverse effects. However, the combination may result in the potential risk of drug-herb interactions. Further clinical studies are needed to evaluate the efficacy and safety of the CHM and WM coprescriptions for FBC patients.

Effects of alfa calcidol and diphosphonates on the BMD of patients with spinal cord injury / 中国康复理论与实践

@#ObjectiveTo study effects of oral administering alfa calcidol and diphosphonates on the bone mineral density (BMD) of patients with spinal cord injury (SCI).MethodsChanges of BMD of the 4th lumbar spine (L4), the proximal femur (total) and all of its subareas were studied between SCI cases of taking (n=31) and not taking (n=42) alfa calcidol. Changes of BMD between cases of taking diphosphonates (n=22) and not taking (n=24) were also compared.ResultsThere were no significant differences in BMD of L4, the proximal femur and all of its subareas between SCI cases of taking and not taking alfa calcidol. While there was a significant difference in BMD of the proximal femur (total) between cases of taking and not taking diphosphonates (P<0.05), and the reducing degree of BMD in the former is less than that in the latter.Conclusion Oral administering diphosphonates has a protective effect on SCI patients' BMD of the proximal femur.