AIMTo study the structure and crystal forms of chlorobenzylidine.

METHODSKarl Fischer titrimetry, FTIR, thermal analysis, single and powder X-ray diffraction were used for the studies of the structure of chlorobenzylidine and for the identification of two forms of chlorobenzylidine.

RESULTSChlorobenzylidine and its diastereoisomer have been studied in this article. They can be distinguished by their different melting points. Two crystal forms of chlorobenzylidine (form A and form B) have also been detected and studied. Form A was studied by single-crystal X-ray diffraction, it crystallized in the triclinic system, space group P1(-), with two formula units per cell, is monohydrate. Karl Fischer titrimetry, FTIR, thermal analysis and powder X-ray diffraction were used for identification of the two forms.

CONCLUSIONThe studies of structure and crystal forms of chlorobenzylidine are very useful for the clinical research and the selection of recrystallization process.

Benzylidene Compounds ; Crystallization ; Crystallography, X-Ray ; Differential Thermal Analysis ; Molecular Conformation ; Molecular Structure ; Polycyclic Compounds ; chemistry ; Stereoisomerism

To compare the DAAO expression level in different Pichia pastoris host strains, the gene encoding DAAO from Trigonopsis variabilis was cloned into plasmid pPIC3.5k and then transformed into P. pastoris GS115 and KM71 respectively. The positive transformants PDK13 (MutS) and PD27 (Mut+) were obtained by PCR analysis. Their optimal and different expression conditions were investigated. To compare with PD27, PDK13 was determined to poss a slower consumption of methanol, a longer induction time, a lower oxygen request and apparently higher expression of DAAO. The highest expression levels were reached up to 2700, 2500 IU/L in shaking flask and 10140, 8463.5 IU/L in fermentor respectively. The over-expression of DAAO can meet its large demand for production of 7-ACA, alpha-keto acid and L-amino acid. In addition, the phenylpyruvate and L-phenylalanine were obtained by crude DAAO reacting with DL-phenylalanine at 37 degrees C for 3h.
D-Amino-Acid Oxidase ; genetics ; Fermentation ; Methanol ; metabolism ; Phenylalanine ; metabolism ; Pichia ; genetics ; Polymerase Chain Reaction

BACKGROUNDDiabetic nephropathy is a common complication of diabetes mellitus. This study aimed to explore whether mesenchymal stem cells (MSCs) transplantation could attenuate diabetic nephropathy in experimental diabetic rats.

METHODSSprague-Dawley rats received a single intraperitoneal injection of streptozotocin (STZ) (60 mg/kg). Diabetic rats were randomized to four groups: diabetes control group (DC), ciclosporin A group (CsA), MSC group, and MSC + CsA group (MSCA). Bone marrow mesenchymal stem cells were cultured, identified and labeled by 5-bromo-2'-deoxyuridine (BrdU) in vitro. Then they were transplanted to diabetic rats via introcardiac infusion. Ciclosporin A was administered daily at 5 mg/kg. At 1, 2, 4, 8 weeks after transplantation, random blood glucose, urine albumin/creatinine ratio (Alb/Cr), endogenous creatinine clearance rate and renal mass index were tested. Renal morphology and labeled cells were examined.

RESULTSCultured MSCs expressed mesenchymal cell phenotype, and could be multidifferentiated to osteogenic and adipogenic cells. Labeled MSCs could be detected in the kidney of nephropathic rats, mainly in renal interstitium, but they did not propagate after engrafting in kidney. Over the course of the experiment, MSCA group showed a significant decrease in blood glucose compared with MSC group, CsA group and DC group (P < 0.05, respectively). The Alb/Cr in MSCA group and MSC group were significantly lower than CsA group and DC group (P < 0.05). And the Alb/Cr in MSCA group showed a significant decrease compared with MSC group (0.74 vs 0.84, P < 0.05). There was a significant difference in renal mass index between the MSCA group and DC group (5.66 vs 6.37, P < 0.05). No significant difference was found in creatinine clearance rate among 4 groups (P > 0.05). Treatment with MSC + CsA significantly ameliorated the morphology of diabetic kidney.

CONCLUSIONMSC could mildly ameliorate diabetic nephropathy by decreasing blood glucose, Alb/Cr ratio and renal mass index.

Animals ; Blood Glucose ; Diabetic Nephropathies ; blood ; metabolism ; therapy ; Flow Cytometry ; Immunohistochemistry ; Kidney ; metabolism ; pathology ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Microscopy ; Rats ; Rats, Sprague-Dawley

Ultra-high performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOF-MS) was applied to rapidly identify the phospholipids in human plasma and explore the mass spectrometric fragmentation pattern. An acquity UHPLC^TM BEH C18 column (50 mm × 2.1 mm, 1.7 μm) was utilized and eluted with a gradient system; the mobile phase consisted of 10 mmol·L^-1 ammonium formate aqueous solution-0.1% formic acid aqueous solution (A) and acetonitrile-isopropanol (1∶1) organic solution (B) containing 10 mmol·L^-1 ammonium formate-0.1% formic acid. The flow rate was 0.3 mL·min^-1 and the column temperature was set at 50 ℃. An electrospray ionization (ESI) source was used to collect mass spectra in positive and negative ion mode. Based on the precise relative molecular weight and elemental composition calculated by Masslynx 4.1 software, comparison with references, and secondary mass spectrometry fragment ions and lipid databases, a total of 82 plasma lipids were identified, including 14 lysophosphatidylcholines (LysoPCs), 39 phosphatidylcholines (PCs), 17 sphingomyelins (SMs), 7 ceramides (Cers), 4 phosphatidylethanolamines (PEs), and 1 phosphatidylinositol (PI). A simple, efficient, fast and stable analytical method was established in this study for the qualitative analysis of phospholipids in human plasma, and the fragmentation regularity of the main phospholipids was determined. This work provides a good foundation for further metabolomics studies of plasma phospholipids. This study was approved by the Second Affiliated Hospital of Guangzhou Medical University Clinical Research and Application Institutional Review Board Approval (No. 2020-hs-07).

Objective To explore olanzapine effect on the cognitive function and neuronal damage of aged schizophrenic rats based on the PI3 K/Akt signaling pathway. Methods Ten-week-old SD rats were randomly divided into a blank control group（n=12） and a modeling intervention group（n=48）. The modeling group were injected with didroxapine maleate [MK-801,0.2 mg/（kg·d）] for 14 days. And the model was evaluated by general behavioral studies to determine the success of model building. The model rats were randomly divided into model group and low, medium, and high dose olanzapine groups [10, 20, 40 mg/（kg·d）], each with 12 rats. The control group and model group were given distilled water; the low, medium, and high dose olanzapine groups were given olanzapine for 21 days. The stereotyped lines were scored by the standard of Sams Dodd and Hoffman, the cognitive evaluation of the rats was performed by the Morris water maze, and the levels of interleukin-6（IL-6） and tumor necrosis factor-α（TNF-α） in the serum were determined by ELISA. The activities of dihydrokaempferol（Ach） and acetyl cholinesterase（AchE）in brain tissue were detected by acetylcholinesterase activity assay kit. Rat brain tissue PI3 K, Akt, mammalian target of rapamycin（mTOR） mRNA expression levels were detected by Real-time PCR. Results Compared with the model group, the stereotyped behavior and ataxia scores, escape latency, number of crossing platforms, serum levels of IL-6, TNF-α, AchE, phosphorylated PI3 K（p-PI3 K）, phosphorylated Akt（p-Akt） protein expression decreased（P＜0.05 or P＜0.01）, while brain tissue Ach, PI3 K, mTOR and phosphorylated mTOR（p-mTOR） protein content increased（P＜0.05 or P＜0.01） in the low, medium and high dose olanzapine groups. The content of Akt was increased in the low-dose group. Compared with the model group, Akt and mTOR mRNA in the brain tissue of rats in the low, medium, and high-dose alanzapine groups expression levels were down-regulated（P＜0.05 or P＜0.01）. PI3 K mRNA in the brain tissue of rats in the low, medium, and high-dose alanzapine groups expression levels were down-regulated（P＜0.05 or P＜0.01）. Conclusion Olanzapine can reduce stereotyped behavior and ataxia scores, escape latency, number of crossing platforms, IL-6, TNF-α, AchE and increase Ach content and regulate the PI3 K/Akt signaling pathway to relieve the schizophrenia.