Improved γ-Linolenic Acid Production from Cellulose in Mucor circinelloides via Coexpression of Cellobiohydrolase and Delta-6 Desaturase.

The present study was aimed at facilitating the production of γ-linolenic acid (GLA) from the cellulosic substrate with the engineered oleaginous fungus Mucor circinelloides WJ11. Here, the homologous recombination technology was used to overexpress the cellobiohydrolase (CBH2) derived from Trichoderma longibrachiatum and the original delta-6 fatty acid desaturase (D6) in M. circinelloides to construct genetically engineered strains capable of effectively using cellulose to enhance GLA synthesis. When cultivated in modified K&R medium supplemented with microcrystalline cellulose, the CBH2 and D6 coexpressing strains led to increases in the biomass (up to 12.8 g/L) and lipid yield (up to 3.7 g/L) of 87% and 2.4-fold, respectively, compared to that of the control strain. Notably, when CBH2 and D6 were coexpressed in M. circinelloides, the yield of GLA reached 608 mg/L, which was a dramatic increase of 3.9-fold compared to that of the control strain. This is the first report on promoting the GLA production from the cellulosic substrate via coexpression of CBH2 and delta-6 desaturase. This work provides a theoretical basis for efficient transformation from the cellulosic substrate to functional GLA by CBH2 and D6 coexpressing strains, which might play a positive role in promoting the sustainable development of biological industry.

[Establishment of quality evaluation methods for pieces and standard decoction of honey-fried Descurainiae Semen].

To establish a content determination method for quality control of the pieces and standard decoction of honey-fried Descurainiae Semen. Standard decoction of honey-fried Descurainiae Semen was prepared with standardized process, and high performance liquid chromatography coupled with diode-array detector(HPLC-DAD) was used to detect its characteristic fingerprint and determine the content of quercetin-3-O-ß-D-glucose-7-O-ß-D-gentiobioside. In addition, the transfer rate, dry extract rate and pH value were calculated. The results showed that the established method had a high accuracy. The content of quercetin-3-O-ß-D-glucose-7-O-ß-D-gentiobioside in 13 batches of standard decoction was 0.03-0.12 mg·mL~(-1); the transfer rate was 13.4%-23.1%; the rate of extracts was 1.9%-5.5%, and the pH was between 5.4-5.9. The similarity coefficients were all greater than 0.85, indicating good homogeneity for the different batches of decoction. There were 7 common peaks in the characteristic chromatogram, one of which was quercetin-3-O-ß-D-glucose-7-O-ß-D-gentiobioside. In this paper, the established content determination and quality evaluation method for Descurainiae Semen pieces and decoction was simple, rapid and reproducible, providing reference for the quality control of honey-fried Descurainiae Semen pieces, standard decoction and its preparations.

Symptomatic treatment of brain metastases in renal cell carcinoma with sorafenib.

Brain metastasis is synchronous to the diagnosis of renal cell carcinoma (RCC). The prognosis of brain metastasis in RCC with the current treatment options is dismissal. Therefore, we present a case of an elderly female patient with RCC showing a partial response of brain metastasis after 18 months of 600 mg once daily sorafenib treatment who underwent right-sided nephrectomy. Further, withdrawal of sorafenib resulted in psychiatric changes along with increased metastasis lesions, which were recovered upon resuming the treatment, proposing that oral sorafenib can be used safely and efficiently for treatment of brain metastasis in advanced RCC.

Flower-Like Nickel Phosphide Microballs Assembled by Nanoplates with Exposed High-Energy (0 0 1) Facets: Efficient Electrocatalyst for the Hydrogen Evolution Reaction.

The fabrication of low-cost and earth-abundant electrocatalysts for the hydrogen evolution reaction (HER) over a broad pH range is attractive. In this work, a facile precursor route is developed to synthesize flower-like nickel phosphide microballs with a diameter of approximately 12 µm. With a controlled phosphorization temperature, flower-like nickel phosphide microballs with different crystalline structures (Ni5 P4 and Ni2 P) were obtained easily. Flower-like Ni5 P4 microballs possessed two advantageous features for enhanced HER: fast vectorial electron transfer path along the building block nanoplates and enhanced inherent catalytic activity of each active site for high-energy (0 0 1) facets. The flower-like Ni5 P4 microballs electrocatalyst thus displayed excellent activity for the HER with a low overpotential (η) of 35.4 mV to reach current densities of 10 mA cm-2 and a small Tafel slope of 48 mV dec-1 in acid solution. In addition, it showed excellent activity in 1 m KOH with η=47 mV at 10 mA cm-2 . DFT studies indicated that the free energy of hydrogen adsorbed on the Ni site of Ni5 P4 was 0.152 eV, which is smaller than that of the Ni site of Ni2 P (0.182 eV). Therefore, flower-like Ni5 P4 microballs exhibited better HER activity than Ni2 P, which is consistent with our HER data. This hierarchical structure with exposed high-energy (0 0 1) facets paves the way to design and synthesize low-cost, high-performance catalysts for the HER.

Cutting edge: Ubiquitin-specific protease 4 promotes Th17 cell function under inflammation by deubiquitinating and stabilizing RORγt.

RORγt is a key transcription factor that controls the development and function of inflammatory Th17. The mechanisms that regulate RORγt stability remain unclear. We report that Th17 cells highly express the deubiquitinase ubiquitin-specific protease (USP)4, which is essential for maintaining RORγt and Th17 cell function. Inhibition of the catalytic activity of USP4 with vialinin A, a compound derived from Chinese traditional medicine, dampened Th17 differentiation. USP4 interacted and deubiquitinated K48-linked polyubiquitination of RORγt, thereby promoting RORγt function and IL-17A transcription. Interestingly, TGF-ß plus IL-6 enhanced USP4-mediated deubiquitination of RORγt. Moreover, USP4 and IL-17 mRNA, but not RORγt mRNA, were significantly elevated in CD4(+) T cells from patients with rheumatic heart disease. Thus, USP4 could be a novel therapeutic target for the treatment of Th17-modulated autoimmune diseases.

Bufalin exerts antitumor effects by inducing cell cycle arrest and triggering apoptosis in pancreatic cancer cells.

As one of the most aggressive human malignancies, pancreatic cancer is a leading cause of cancer-related deaths worldwide and only about 4% of patients will live 5 years after diagnosis. Eighty to approximately eighty-five percent of patients are diagnosed with an unresectable or metastatic disease, which is correlated with poor prognosis and low survival rate. Therefore, it is tremendously significant to exploit novel chemicals to prevent and treat pancreatic cancer. Previous research and clinical studies have demonstrated that many natural products derived from traditional Chinese medicine (TCM) such as camptothecin derivatives and vinca alkaloids could be effective antitumor compounds, hinting that TCM is a promising source for developing new antitumor drugs. In this report, we investigated the effects of bufalin, a primary active ingredient of the traditional Chinese medicine Chan-Su, on pancreatic cancer cell lines PANC-1 and CFPAC-1 and studied the underlying molecular mechanism. We found that exposure to bufalin could suppress the proliferation of pancreatic cancer cells time and dose dependently. We used flow cytometry to study the effects of bufalin on apoptosis and cell cycle distribution in PANC-1 and CFPAC-1 cells. The results indicated that bufalin could significantly induce both apoptosis and G2/M cell cycle arrest in pancreatic cancer cells. With western blotting, we found that the expression level of an antiapoptotic protein heat shock protein 27 (Hsp27) and its partner molecule p-Akt was decreased upon the treatment with bufalin. Besides, bufalin activated pro-caspase-3 and pro-caspase-9 and modulated the expression level of Bcl-2 and Bax. These data suggested that bufalin may trigger apoptosis by targeting Hsp27, which could inhibit apoptosis by interfering with key apoptotic proteins. The influence on the level of cylinB1, CDK1, and p21 was also observed after bufalin treatment, and the relationship between Hsp27 and the cell cycle-related proteins mentioned above deserves much more research. In addition, our data showed that bufalin could enhance the growth inhibition effect of gemcitabine in above pancreatic cancer cells. Taken together, bufalin might be worthy of further study for its potential as a therapeutic agent for pancreatic cancer treatment.

Concurrent chemoradiotherapy with or without adjuvant chemotherapy in intermediate and locoregionally advanced nasopharyngeal carcinoma.

Concurrent chemoradiotherapy (CCRT) showed a significant improvement in disease control and clinical outcome in patients with intermediate and locoregionally advanced nasopharyngeal carcinoma (NPC) (stage II, III and IVA+B). However, there has been debate about the contribution and application of additional adjuvant chemotherapy (AC) to a CCRT regime. This study aims to evaluate the additional value of AC in the treatment of intermediate and locally advanced NPC with regard to toxicity and clinical outcomes. A total of 189 patients with American Joint Committee on Cancer (AJCC) stage II to stage IVB NPC were retrospectively identified. Patient characteristics, toxicity, compliance with treatment and clinical outcomes, including response to treatment, overall survival (OS), progression-free survival (PFS), relapse-free survival (RFS), freedom from local recurrence (FLR) and freedom from distant metastasis (FDM), were analyzed. The overall response rate of CCRT and CCRT/AC groups was 97.92 % and 97.83 %, respectively (P=0.643). The 5-year OS rate was 68.2 % in the CCRT group and 75.9 % in the CCRT/AC group (P=0.53). The 5-year PFS rate was 66.7 % and 71.4 % in CCRT and CCRT/AC groups, respectively (P=0.96). This study showed no evidence of an additional value of AC in CCRT treatment in disease control and clinical outcomes in patients with locally advanced NPC in endemic regions. Moreover, three additional cycles of AC after CCRT appeared to be poorly tolerated in patients. Therefore, AC should not be routinely used for treatment, although clinical trials may be justified.

[Effects of Radix Ophiopogonis decoction on embryo-fetal development in rats].

OBJECTIVE: To investigate the potential developmental toxicity of Radix Ophiopogonis decoction in SD rats. METHOD: Timed-pregnant SD rats were given Radix Ophiopogonis decoction (26.9 g x kg(-1)) or vehicle (distilled water) by gavage on gestation days 6-17. Maternal clinical sign, abortions, premature deliveries, and body weight were monitored throughout gestation. At termination (gestation days 20) pregnant females were evaluated for clinical status and gestational outcome; live fetuses were examined for gender, external, visceral and skeletal malformation and variations. RESULT: No deaths, premature deliveries or dose-related clinical signs were attributed to Radix Ophiopogonis decoction. Maternal body weight and body weight gain were not affected. There were no effects on fetus weight and viability, incidences of fetal malformation and variation. CONCLUSION: These results demonstrated that Radix Ophiopogonis decoction had no detectable adverse effects in either the treated F0 female rats or the fetuses.

[Genotoxicity study of Cortex Fraxini decoction].

OBJECTIVE: To investigate the genotoxicity of Cortex Fraxini decoction. METHOD: Mouse lymphoma assay (MLA) and mouse bone marrow micronucleus test (MNT) were used. In MLA, four dose levels of 1.71, 3.42, 6.83 and 13.65 g (crude drug) x L(-1) were exposed with I5178Y cells for 3 hours with and without metabolic activation, then expressed for 2 days. The mutation frequency plates were prepared and incubated for 12-13 days. Colony size in each plate was scored, and the total mutation frequency and the percentage of small colony mutants were calculated. In MNT, contained three dose levels of 7.14, 14.28 and 28.55 mg (crude drug) x kg(-1) and 10 ICR mice (5 males/5 females) were in each group. The mice were given in every 24 hours by oral gavage twice and sacrificed after 24 hours of the last dosing. Both femur bones were collected to prepare the smear. For each mouse, the number of micronucleated polychromatic erythrocytes (MNPCE) in 2 000 polychromatic erythrocytes was counted, and the mean of rate of MNPCE of each group was calculated. RESULT: In MLA, the results indicated that the total mutation frequency of four dose levels showed a dose-dependent increase, there was statistically significant difference at high concentrations compared with negative control (P < 0.01), and the percentage of small colony mutants was similar with positive control in the absence of metabolic activation. The total mutation frequency of each dose level was similar with negative control in the presence of metabolic activation. In MNT, the results indicated that the decoction did not show inhibitory action for bone marrow, and the induced rate of MNPCE of each group was not significantly increased comparing with negative control. CONCLUSION: Cortex Fraxini decoction induces the tk(+/-) gene mutation and chromosome damage in L5178Y cells in vitro without metabolic activation, it hints that the direct mutagens may be within the test article. Cortex Fraxini decoction does not show chromosome damage of bone marrow in ICR mice, it has not genotoxicity in vitro/in vivo with metabolic activation under this study condition.