Objective To observe effect of TNF-? on PPAR?2 mRNA expression in 3T3-L1 cells transfected with human recombinant adiponectin. Methods The 3T3-L1 preadipocytes were transfected with the recombinant plasmid pcDNA3.1+-hADPN.The PPAR?2 mRNA expression was quantitated by semi-quantitative RT-PCR. Results (1)Compared with controls(the 3T3-L1 cells and the 3T3-L1 cells with plasmid), the PPAR-?2 mRNA expression of 3T3-L1 cells with human recombinant adiponectin was higher(P

Aged ; Coal Mining ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; diagnosis ; etiology ; Pneumoconiosis ; complications

Olfactory neuroblastoma is a rare malignant tumor. Although multiple therapeutic modalities including surgery, radio-therapy, or chemotherapy could be used in patients with olfactory neuroblastoma, no standardized treatment has been achieved. This re-view introduces a case of adult olfactory neuroblastoma treated by a multiple disciplinary team in Tianjin Medical University Cancer In-stitute and Hospital. This review also aims to explore a complete set of diagnostic and treatment practices for the benefit of future pa-tients.

Objective: To explore the effect of single-nostril transsphenoidal approach on pituitary adenoma. Methods: We retrospectively analyzed 46 cases of pituitary tumors treated with single-nostril transsphenoidal approach and the effects and complications of surgery.Dunng the surgery,a nasal speculum was inserted through right nostril slowly towards the anterior wall of sphenoid sinus.A nasal mucosa incision of about 1.5cm was made in the right nasal cavity at the level of the middle nasal turbinate.With a fracture of the bony septum,a space was developed between the bilateral nasal mucosa and bony septum to the sphenoid sinus.Then,the face of the sphenoid sinus was exposed.The remainder of the bony septum,the anterior sphenoid sinus wall,and the sphenoid mucosa were removed.The antenor sphenoidotomy should be less than 1.5cm wide.After confirming the tumor by dural puncture,a cross incision of dura was made and the tumor was removed.The saddle was usually Collapsed and visible after total tumor removal.When the tumor was resected,sevaral gelatin sponges were stuffed into the Surgical cavity to stop bleeding. Results: Thirty-four cases had total resection and 12 cases had subtotal resection.No deaths or disability occurred.Hormone levels in almost all patients were improved.Seventeen cases had a sign of diabetes insipidus.Electrolyte disturbance occuwed in 5 cases.NO postoperative cerebrospinal fluid rhinorrhea was observed. Conclusion: Single-nostril transsphenoidal approach has many advantages in treating pituitary adenomas such as simplified approach,brief technology and high security.

Clinical laboratory biological safety is one of whole society safety. This paper introduced briefly the current situation of clinical medical laboratory biosafty in the hospital. and set forth common biological hazards specifically for whose characteristics. Combining the biosafety administration measures from abroad, the issue of laboratory biological safety administration was considered, and put forward some suggestions according to related law and regulation of national laboratory safety administration in order to strengthen clinical laboratory biosafety administration.

Glucose-6-phosphate dehydrogenase is main regulatory enzyme for pentose phosphate pathway. To amplify the core sequence of G6PDH gene from Chimonanthus praecox, the primers were synthesized, based on the conserved nucleotide sequence of other reported plant G6PDH genes. The specific primers were designed according to the major fragment. The full length cDNA of the G6PDH1 gene was isolated by the 3' and 5' rapid amplification of cDNA ends approach. Transcript levels of G6PDH1 isoform was measured by real-time quantitative RT-PCR in different tissues and in responds to cold treatment. The G6PDH1 subcellular localization, transmembrane domain, three-dimensional structure, and phylogenetic analysis were predicted by different software to analysis the bioinformatics of G6PDH1 protein. The G6PDH1 cDNA sequence was 2 011 bp in length and consisted of 1 551 bp Open Reading Frame (ORF) , encoding a protein of 516 amino acids. Expression analysis results in different tissues showed that G6PDH1 was primarily observed in flowers and roots, as opposed to the leaves and stems. Cold treatment experiments indicated that cold treatment caused a rapid increase in G6PDH1 expression in flowers within 12 h. The full-length cDNA of G6PDH1 and its expression analysis will play an important role for further study on cold stress responses in Ch. praecox.
Calycanthaceae ; chemistry ; classification ; enzymology ; genetics ; Cloning, Molecular ; Enzyme Stability ; Glucosephosphate Dehydrogenase ; chemistry ; genetics ; metabolism ; Models, Molecular ; Open Reading Frames ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism

Lysine decarboxylase is a key enzyme involved in the upstream biosynthesis of lycopodium alkaloids (LAs) such as huperzine A, contributing to the decarboxylation of lysine to 1,5-pentanediamine (cadaverine). Three lysine decarboxylase genes (HsLDC-L1, HsLDC-L2, HsLDC-L3) were successfully cloned from Huperzia serrata using transcriptomic sequence data mining strategy combined with reverse transcription PCR. The physicochemical properties, secondary and tertiary structures, amino acid identities, and evolutionary relationship of the three LDCs were analyzed by online bioinformatics analysis platforms and DNAMAN, MEGA 7.0 software, revealing that all of these proteins had the conserved PLP binding domain and active site residues were completely conserved in LDCs. Phylogenetic analysis showed that these LDCs were located in the same branch as other known LDCs from LA-producing plants. Accordingly, the ORFs of these three HsLDCs were inserted into different expression plasmids for further expression in E. coli. However, only HsLDC-L1 was successfully expressed in E. coli BL21 (DE3) by inserting into a pCold TF vector. The recombinant protein was purified by Ni²⁺ affinity chromatography purification. HsLDC-L1 contains 469 amino acid residues, with a calculated molecular weight of 50.50 kDa. HsLDC-L1 expectedly catalyzed the decarboxylation of lysine to produce cadaverine. In addition, HsLDC-L1 can also catalyze the generation of putrescine from ornithine. However, it cannot catalyze the decarboxylation of tyrosine, phenylalanine, tryptophan and histidine. The results not only provide insight into the biosynthesis of LAs including huperzine A, but also provide a critical genetic element for the overproduction of Δ¹-piperideine and pelletierine, the essential biosynthetic precursors of LAs, using synthetic biology strategies.

Objective To lay the foundation for studying the possible pathogenesis of epilepsy and the anti-epileptic mechanism of Banxia Baizhu Tianma Decoction through the bioinformatic analysis of target gene prediction and signal pathway of miRNA-146a-5p in hippocampus of epileptic rats. Methods Lithium-pilocarpine was used to induce seizures in rat models. The experiment rats were randomly divided into normal control group, model group, Banxia Baizhu Tianma Decoction group, with 20 rats in each group. The method of miRNA expression profiling was used to observe the miRNA differential expression of hippocampus neuron cell of rats. The expression level of miRNA-146a-5p was detected by real-time quantitative PCR. MiRDB was used for target gene prediction of miRNA-146a-5p, and miRTarBase and DAVID were used for enrichment analysis on the GO function and KEGG signaling pathway. Results The attack times and grades of the rats in Banxia Baizhu Tianma Decoction group were significantly lower than those in the model group from behavioral observation. MiRNA microarray analysis showed that the expression level of miRNA-146a-5p in model group was 2.107 times normal control group (P＜0.05), and the expression level decreased to 1.377 times after treatment with Banxia Baizhu Tianma Decoction (P＜0.05). The results of RT-PCR was consistent with that of miRNA microarray, with statistical significance (P＜0.05). MiRNA-146a-5p target gene prediction results had 140 target genes by GO, and there were 14 annotation information of biological process (P＜0.05), 9 annotation information of cellular component (P＜0.05), 11 annotation information of molecular function (P＜0.05). Enrichment analysis of KEGG biological pathway showed that 140 target genes of miRNA-146a-5p were enriched in EB virus infection signal pathway and thyroid hormone signaling pathway (P＜0.05). Conclusion miRNA-146a-5p is closely related to the inflammatory reaction after epilepsy, and Banxia Baizhu Tianma Decoction can control epilepsy possibly by controlling the inflammatory reaction after epilepsy.

BACKGROUNDThe fat derived protein adiponectin plays an important role in the regulation of glucose metabolism. The aim of this study was to provide the experimental basis for further investigating on adiponectin (ADPN) function. Its eukaryotic recombinant was constructed and expressed in precursor cells of 3T3-L1 adipocytes. The effects of dexamethasone on peroxisome proliferator activated receptor-gamma (PPAR-gamma) mRNA expression in 3T3-L1 cells with human recombinant adiponectin were assessed.

METHODSThe recombinant plasmid pMD18-T-hADPN and eukaryotic expression vector pcDNA3.1(+) were digested by two restrictive endonucleases and adiponectin and linear pcDNA3.1(+) were obtained. Then, they were ligated and translated into JM109. The recombinant pcDNA3.1(+)-hADPN so obtained was identified by digestion by restrictive endonuclease and nucleotide sequencing. The 3T3-L1 precursor cells were transfected using SuperFect Transfection Reagent (Qiagen). Furthermore, 3T3-L1 cells with human recombinant adiponectin incubated with dexamethasone (0.5 mmol/L) for 24 hours, cells were collected and total RNA was extracted. The PPAR-gamma mRNA expression was quantified by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSAfter eukaryotic recombinant was digested by Hind III and EcoR I, fragments of 800 bp and 5.4 kb were identified by nucleotide sequence scanning and consistent with theoretical values. Electrophoretogram of RT-PCR in 3T3-L1 precursors showed only one band in front of 250 bp, which was consistent with theoretical value 234 bp. In the 3T3-L1 cells, 3T3-L1 cells with plasmid and 3T3-L1 cells human recombinant adiponectin, treatment with dexamethasone (0.5 mmol/L) decreased PPAR-gamma mRNA expression compared to untreated controls (P < 0.01). Effect of dexamethasone on PPAR-gamma mRNA expression in 3T3-L1 cells was reversed by stably transfected human recombinant adiponectin.

CONCLUSIONThe 3T3-L1 cells stably transfected human recombinant adiponectin had increased PPAR-gamma mRNA expression. Dexamethasone suppressed PPAR-gamma mRNA expression in the 3T3-L1 cells. Effect of dexamethasone on PPAR-gamma mRNA expression in 3T3-L1 cells was reversed by stably transfected human recombinant adiponectin.

3T3-L1 Cells ; Adiponectin ; physiology ; Animals ; Dexamethasone ; pharmacology ; Insulin Resistance ; Mice ; PPAR gamma ; genetics ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction

AIMTo study the chemical constituents of Selaginella tamariscina (Beauv.) Spring.

METHODSVarious chromatographic techniques were used to separate and purify the chemical constituents. Their physico-chemical properties and spectral data were used to elucidate the structures.

RESULTSFour compounds were isolated from the n-BuOH fraction of the water-extracts. Their structures were identified as 1-hydroxy-2-[2-hydroxy-3-methoxy-5-(1-hydroxyethyl)-phenyl]-3-(4-hydroxy-3,5-dimethoxy)-propane-1-O-beta-D-glucopyranoside (tamariscinoside B, I), adenosine (II), guanosine (III), arbutin (IV).

CONCLUSIONTamariscinoside B (I) is a new compound, while the others were isolated from Selaginella for the first time.

Adenosine ; chemistry ; isolation & purification ; Arbutin ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Guanosine ; chemistry ; isolation & purification ; Molecular Conformation ; Molecular Structure ; Plants, Medicinal ; chemistry ; Selaginellaceae ; chemistry