Secondary Metabolite Analysis of Phomopsis sp. LGT-5 Based on the Dual Guidance of Whole-Genome Sequencing and HPLC-Q-ToF-MS/MS.

Microorganisms produce a wealth of structurally diverse specialized metabolites with a remarkable range of biological activities. The Phomopsis sp. LGT-5 was obtained through tissue block and repeatedly crossed methods from Tripterygium wilfordii Hook. F. The antibacterial experiments of LGT-5 showed that it has high inhibitory activity against Staphylococcus aureus and Pseudomonas aeruginosa, and moderate inhibitory activity against Candida albicans. To research the generation of the antibacterial phenomenon of LGT-5 and provide support for further research and application, the whole genome sequencing (WGS) of LGT-5 was obtained by single-molecule real-time DNA sequencing platform Pacific Biosciences (PacBio) sequencing and Illumina paired-end sequencing. The final assembled LGT-5 genome is 54.79 Mb with a contig N50 of 290.07 kb; in addition, its secondary metabolites were detected through HPLC-Q-ToF-MS/MS. By comparing its MS/MS data, the secondary metabolites were analyzed based on visual network maps obtained on the Global Natural Products Social Molecular Networking (GNPS). The analysis results showed that the secondary metabolites of LGT-5 were triterpenes and various cyclic dipeptides.

miR-204-5p Plays a Critical Role in the Pathogenesis of Depression and Anti-depression Action of Venlafaxine in the Hippocampus of Mice

Background: Background: Venlafaxine has been demonstrated to treat diseases such as social anxiety disorder and depression. Most of antidepressants including venlafaxine have a certain effect, but significant side effects. Therefore, it is necessary for us to research the development of novel antidepressants for effective treatment in practice. MicroRNA-204 (miR-204) is highly expressed in brain tissue, and plays a critical role in the synaptic plasticity of hippocampal neurons in rats. However, the underlying molecular mechanism of miR-204 remains unclear to date, this study aims to offer unique insights into depression and provide a theoretical basis for clinical physicians. Methods: A chronic social defeat stress (CSDS) was initially adopted for establishing a mice model of depression in this research and depression-like behaviors were evaluated by a series of behavioral experiments including the sucrose preference test (SPT), the tail suspension test (TST), the forced swim test (FST) and the social interaction test (SIT). Quantitative real-time reverse transcription PCR (qRT-PCR) was also conducted to test the expression levels of miR-204 and BDNF in the hippocampus of mice. Finally, gene interference of miR-204-5p was further adopted to test whether miR-204-5p played an effective role in the antidepressant effects of venlafaxine in mice. Results: Our data implicated that CSDS significantly increased the miR-204-5p but not miR-204-3p levels in the hippocampus of mice. The treatment of venlafaxine obviously relieved depression- like behaviors of CSDS-induced mice. The usage of venlafaxine abolished the increasing effects on the expression of miR-204-5p but up-regulated the BDNF expression level in CSDS-exposured mice. More importantly, we found that genetic overexpression of miR-204-5p decreased the reverse effects of venlafaxine on depressive-like behaviors and genetic knockdown of hippocampal miR-204-5p relieved the depressive-like behaviors and neurogenesis in CSDS-induced mice. Conclusion: miR-204-5p played an effective role in the antidepressant effects of venlafaxine in CSDS-induced mice.

Highly Active La(III)-Based Metal-Organic Framework as a Heterogeneous Lewis Acid Catalyst for Friedel-Crafts Alkylation.

In this study, a novel La(III)-based two-dimensional (2D) metal-organic framework, [La2/3 (qptca)1/2 ] (referred to as SLX-2), from LaCl3 and 1,1' : 4',1'' : 4'',1''' : 4''',1''''-quinquephenyl]-2,2'',2'''',5''-tetracarboxylic acid (H4 qptca) was synthesized by conventional solvothermal method and thoroughly characterized by using X-ray single-crystal diffraction, powder X-ray diffraction, and thermogravimetric analyses. The 2D SLX-2 features a unique lanthanum center exposed to the skeleton and was used as an efficient Lewis acid catalyst for the Friedel-Crafts alkylation of indole and pyrrole with ß-nitrostyrene along with a wide substrate scope, giving the desired products in good-to-high yields under the optimal reaction conditions. Furthermore, the catalyst was used for twenty cycles, with nearly no effect on its activity, and the reaction was heterogeneous in nature. Moreover, compared to the previous hydrogen-bond-donating MOF catalysts for such alkylation reactions, SLX-2 showed an excellent stability toward harsh acidic and basic environment, and gave comparable catalytic activities.

The requirement of phosphoenolpyruvate carboxykinase 1 for angiogenesis in vitro and in vivo.

We here examined the potential biological function of phosphoenolpyruvate carboxykinase 1 (PCK1) in angiogenesis. shRNA- or CRISPR-Cas9-induced PCK1 depletion potently inhibited endothelial cell proliferation, migration, sprouting, and tube formation, whereas ectopic PCK1 overexpression exerted opposite activity. In HUVECs, Gαi3 expression and Akt activation were decreased following PCK1 depletion, but were augmented by ectopic PCK1 overexpression. In vivo, retinal expression of PCK1 gradually increased from postnatal day 1 (P1) to P5. The intravitreous injection of endothelial-specific PCK1 shRNA adenovirus at P1 potently inhibited the radial extension of vascular plexus at P5. Conditional endothelial knockdown of PCK1 in adult mouse retina increased vascular leakage and the number of acellular capillaries while decreasing the number of RGCs in murine retinas. In diabetic retinopathy patients, PCK1 mRNA and protein levels were up-regulated in retinal tissues. Together, PCK1 is essential for angiogenesis possibly by mediating Gαi3 expression and Akt activation.

A New Anti-Inflammatory Alkaloid from Roots of Heracleum dissectum.

Using various chromatographic methods, a new piperidinone alkaloid, (3S)-3-{4-[(1E)-3-hydroxyprop-1-en-1-yl]-2-methoxyphenoxy}piperidin-2-one (1), together with 10 known compounds, bergapten (2), xanthotoxol (3), isopimpinellin (4), isobergapten (5), heratomol-6-O-ß-d-glucopyranoside (6), scopoletin (7), apterin (8), 3-methoxy-4-ß-d-glucopyranosyloxypropiophenone, (praeroside; 9), tachioside (10) and coniferin (11), were isolated from roots of Heracleum dissectum Ledeb. Their structures were elucidated on the basis of physicochemical properties and the detailed interpretation of various spectroscopic data. All the isolated compounds were screened for anti-inflammatory activity in vitro. As the results, compound 1 and 8 showed significantly inhibitory activity on nitric oxide production in RAW264.7 cells.

[Influence of diethyl sulfate (DES) mutagenesis on growth properties and pigment secondary metabolites of Phellinus igniarius].

The diethyl sulfate (DES) mutagenesis was chosen for the mutagenic treatment to Phellinus igniarius, and the relationship of mutagenesis time and death rate was investigated with 0.5% DES. The differences of mycelial growth speed, liquid fermentation mycelia biomass, morphology and pigment classes of secondary metabolites production speed and antioxidant activities of metabolite products were discussed. The study displayed that DES mutagenesis could change mycelial morphology without obvious effect on mycelium growth, and the DES mutagenesis improved antioxidant activities of the active ingredients of P. igniarius and had more antioxidant activity of hypoxia/sugar PC12 nerve cells than that of P. igniarius.

[Expression and clinical significance of DAPK1 and CD147 in esophageal squamous cell carcinoma].

OBJECTIVE: To explore the amplification and expression status of DAPK1 and CD147 in esophageal squamous cell carcinoma (ESCC) and their relationship with the prognosis of ESCC. METHODS: Immunohistochemical staining and RT-PCR were used to detect the expression and amplification of DAPK1 and CD147 in esophageal squamous carcinoma tissue and normal esophageal mucosa. Statistical analysis of the clinocopathological data was performed with SPSS 11.5 software package. RESULTS: The positive rates of expression of DAPK1 protein and CD147 protein in the specimens of esophageal carcinoma were 31.3% and 58.5%, and in normal esophageal mucosa 57.5% and 25.0%, respectively, with a statistically significant difference (P < 0.001). The expressions of DAPK1 and CD147 were significant correlated with invasion depth, lymph node metastasis, TNM stage and the degree of cancer differentiation. (P < 0.05). The negative expression of DAPK1 and positive expression of CD147 indicated a poor prognosis. In 52 ESCC cases, the expression of DAPK1 in cancer tissues was 0.236 ± 0.049, and 0.395 ± 0.058 in normal esophageal mucosa, while that of CD147 mRNA expression was 0.942 ± 0.204 and 0.821 ± 171, respectively, statistically both with a very significant difference (P < 0.01). There was a higher expression level of DAPK1 mRNA in the cancer tissue in patients with no lymph node metastasis, well differentiation, and earlier pathological stage, and a higher expression level of CD147 mRNA in the cancer tissues in patients with lymph node metastasis, poor differentiation, and later pathological stage. CONCLUSIONS: The expression of DAPK1 and CD147 proteins is closely correlated with the clinicopathological characteristics of ESCC. The genes DAPK1 and CD147 may participate in the metastasis and apoptosis of ESCC. The expression of DAPK1 and CD147 may be used as important prognostic predictors in ESCC.

[Influence of different procedures of alimentary tract reconstruction after total gastrectomy for gastric cancer on the nutrition and metabolism of patients: a prospective clinical study].

OBJECTIVE: To investigate the influence of different alimentary tract reconstruction procedures after total gastrectomy for treatment of gastric cancer on nutrition and metabolism and explore an ideal reconstruction procedure. METHODS: A total of 149 patients with gastric cancer who had undergone total gastrectomy were randomly allocated into 5 groups of 30 patients (except the group HLD with 29 cases) to undergo 5 different alimentary tract reconstruction procedures: simple esophagojejunostomy using Roux-en-Y technique (RY), P pouch with Roux-en-Y reconstruction (PRY), jejunal pouch reconstruction according to Hunt-Lawrence technique (HL), jejunal pouch original interposition reconstruction modified by the authors (JOP), and Hunt-Lawrence reconstruction technique maintaining duodenal passage (HLD). Three and six months after operation, quality of life (Visick grade), PNI; body weight; and serum nutritional parameters, including albumin, (ALB), total protein (TP), transferrin (TF), hemoglobin (HB), and serum iron (SI), were evaluated. RESULTS: In comparison with those of the PRY, HL, JOP, and HLD groups, the patients of the RY group show greater weigh loss, and lower ALB, TP, and TF (all P < 0.05). The HB, SI, and TS levels in the JOP group and HLD group were significantly higher than those in the RY, PRY, and HL groups (all P < 0.05). CONCLUSION: Different procedures of alimentary tract reconstruction after total gastrectomy have great influence on the patients' nutrition at different degrees. The patients undergoing the procedures with a reservoir show higher serum nutritional parameters and better body weight. The volume of reservoir has no major clinical importance. The jejunal pouch original interposition reconstruction modified by the authors (JOP), constructing a gastric reservoir and maintaining the alimentary tract flowing through the duodenum is an ideal choice for the reconstruction after total gastrectomy.

[Therapeutic effects of operation in tumor invades adjacent structures cancer of cardia and stomach fundus].

OBJECTIVE: To study the best style of operation in the treatment of tumor invades adjacent structures (T(4)) cancer of the cardia and stomach fundus. METHODS: Two hundred and one patients with T(4) cancer of the cardia and stomach fundus underwent operation. Of them, 31 were treated by laparotomy, and 170 by combined resection of the involved organs. The 3- and 5-year survival rates and the postoperative complication rate and mortality rate were analyzed in the patients who had under gone combined resection of the involved organs. RESULTS: The median survival of the patients undergoing combined resection of the involved organs (29.3 months) was significantly longer than that of those receiving laparotomy (4.9 months). The 3- and 5-year survival rates of 170 patients who had under gone combined resection of the involved organs were 46.2% and 22.8%, respectively. The 3- and 5-year survival rates of patients undergoing total gastrectomy and proximal gastrectomy were 54.9% and 29.2% and 32.2% and 12.5%, respectively, and the difference was statistically significant (chi(2) = 7.589, P < 0.01;chi(2) = 5.792, P < 0.05).The postoperative mortality rate and complication rate were 4.1% and 24.1%, respectively. CONCLUSIONS: The patients without liver metastasis, widespread nodal involvement, peritoneal dissemination and local focus allowed by an en bloc combined resection in T(4) cancer of cardia and stomach fundus should undergo gastrectomy with a combined resection of the involved organs. Total gastrectomy should be performed to improve the curative effect.

[A prospective randomized study on the method of reconstruction after total gastrectomy].

OBJECTIVE: To find an ideal reconstruction method after total gastrectomy. METHODS: With 12 healthy persons as control, a total of 120 gastric cancer patients received their digestive tract reconstruction after total gastrectomy were randomized into Roux-en-y esophagojejunostomy group (A), P pouch with Roux-en-y esophagojejunostomy group (B), Hunt-Lawrence esophagojejunostomy group (C), and jejunal interposition esophagojejunostomy group (D). After operation, quality of life, prognosis nutrition index (PNI), body weight, serum nutritional parameters, gastrointestinal hormone level and immunological state were evaluated. RESULTS: The quality of life, PNI, body weight and serum nutritional parameters (SI, TS and Hb) were better in group D than those in groups A, B and C (P < 0.05). The cholecystokinin (CCK) level and NK cell, CD(4)(+) cell, CD(8)(+) cell and CD(4)/CD(8) ratio in group D, being similar to the control group, were significantly higher than groups A, B and C (P < 0.05). CONCLUSION: Modified jejunal interposition esophagojejunostomy is a reasonable reconstruction method. The construction of "P" pouch, reserving foods as the stomach, can preserve the duodenal passage and secretion of the gastrointestinal hormones, which results in better digestion of the food and absorption of the nutrients. This method simplifies the operation and guarantee the blood supply of interpositioned jejunum without causing ischemia at the anastomotic orifice.

[Long-term therapeutic effects of total gastrectomy in cancer of the cardia and stomach fundus].

OBJECTIVE: To evaluate total gastrectomy for the treatment of cancer of the cardia and stomach fundus. METHODS: Five hundred and thirteen patients with cancer of the cardia and stomach fundus underwent radical resection. Of them, 326 were treated using total gastrectomy (group TG); and 187, using proximal gastrectomy (group PG). The 5-year and 10-year survival rates and the postoperative complication rate and mortality rate were followed up and compared in the two groups. RESULTS: The 5-year and 10-year survival rates of group TG were 43.6% and 24.5%, of group PG were 33.9% and 14.1%, respectively, and the difference was statistically significant (chi(2) = 4.421, P < 0.05, chi(2) = 5.726, P < 0.05). The postoperative complication rate and mortality rate of group TG were 14.7% and 3.1%, of group PG were 10.2% and 2.1%, respectively, and the difference was not statistically significant (chi(2) = 1.796, P > 0.05, chi(2) = 0.082, P > 0.05). CONCLUSIONS: To improve long-term therapeutic effects, total gastrectomy should be recommended for stage III patients with cancer of the cardia and stomach fundus when tumor size is bigger than 3.0 cm or lymph node metastasis occur. The postoperative complication rate and mortality rate should not be increased and the esophagitis of gastroesophageal reflux should be avoided in the patients treated using total gastrectomy.