A new flavonoid glycoside, (-)-2S-8-methyl-5,7,4'-trihydroxyflavanone-7-O-β-D-glucopyranoside (1), along with five known ones, quercetin-3-O-(2"-galloyl)-α-L-arabinoside (2), kaempferol-3-O-α-L-arabinoside (3), guaijaverin (4), trifolin (5) and hyperin (6), was isolated from the leaves of Eucalyptus robusta. Their structures with absolute configurations were elucidated by NMR, HR-ESI-MS, CD spectra data and physicochemical methods. In addition, 2-6 were isolated from E. robusta for the first time.
Eucalyptus ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Plant Leaves ; chemistry

Objective To investigate effects of problem-based learning (PBL) and traditional learning (TL) on students' long-term memory and clinical practice ability. Methods Totally 79 5-year-program undergraduates of 2006, 2007, 2008 grade in school of clinical medicine of our hospital were randomly divided into PBL group (n=38) and TL group (n=41). The teaching effects were evaluated by two exams as well as teachers' subjective impression. SPSS 17.0 statistical analysis software was used;exam results were expressed as x±s; t test and rank sum test were used to analyze the exam results and subjective impression. α=0.05 was set as inspection level. Results In the second exam after 6 months, the mean exam scores were (76.66 ±5.94) and (73.59 ±5.74) in PBL group and TL group, without significant differences between the two groups (t=1.85, P=0.068). However, at clinical intern-ship stage, PBL group outperformed TL group based on the subjective evaluation (P=0.065, 0.277). Conclusion PBL can culture students' ability of problem-solving, but it is limited in culturing long-term memory.

Objective To establish chronic stress model of depression in adolescent rats and to examine the effects of different antidepressant treatment on depression and anxiety-related behaviors.Methods Male Wistar rats were given 21-day chronic mild stress (CMS) during their adolescence (postnatal day 30～50, PND30～50).During stress period, rats were treated with fluoxetine (10 mg/kg i.p.) or imipramine (10 mg/kg i.p.), respectively.After stress, rats were tested for behavioral observation using body weight gain, saccharine test, open field and elevated plus-maze (EPM).Results Compared with control/vehicle (n=10) group, stress/vehicle (n=11) group displayed lower weight gain, saccharine preference index and the number of rearing in open field (P<0.05).Antidepressant fluoxetine, but not imipramine reversed anhedonia and the decrease of the number of rearing induced by stress.In addition, compared with early adolescent(PND29) rats, late adolescent (PND52) rats in control/vehicle group exhibited less open arm entries and open arm time, more closed arm time in EPM (P<0.05).Rats in stress/vehicle group showed more open arm entries and less closed arm time than controls(P<0.05).Both fluoxetine and imipramine had no effects on such changes.Conclusions Stress can induce the depression-like behavior in adolescent rats.Fluoxetine, but not imipramine,can effectively reverse anhedonia induced by stress.However, Both antidepressants have no significant effects on stress-induced decrease in developmental increment of anxious behavior during adolescence.These data suggest that chronic mild stress have complicated effects on depressive and anxious behavior in adolescent rats.

Objective To investigate the expression of cyclin dependent kinases 5(CDK5)in the temporal lobes of the epilepsy patients and to explore the possible roles of CDK5 in the pathogenesis of refractory epilepsy.Methods The brain tissues of intractable epilepsy(IE)were studied by fluorescence quantative polymerase chain reaction(FQ-PCR)for CDK5 mRNA,while immunohistochemistry and Western blot were used to study the protein expression.Nonepileptogenic control brain tissues were used for comparison.Results FQ-PCR analysis showed that the expression of CDK5 mRNA in epilepsy patients was significant higher than those in the control group.And immunohistochemistry showed that the protein mainly existed in the neuron and glial.At the 35000 relative molecular mass,Western blot could been seen that there is a limpid strap.The optical density of CDK5 in IE(temporal lobe 1.4293?0.1839,hippocampus 2.0733?0.4738)was significantly higher than that in the control(temporal lobe 0.9680?0.4147, hippocampus 1.4030?0.6160,P

A recombinant transfer vector, pBacIL-11, containing hIL-11 cDNA of 546 nucleotides lacking leader sequence was constructed and co-transfected into BmN cells with linearized BmBacPAK(modified BmNPV) DNA for construction of a recombinant baculovirus carrying the hIL-11 gene. Southern hybridization analysis suggested that the recombinant baculovirus DNA contained hIL-11 cDNA fragment. RNA dot blotting demonstrated that the hIL-11 gene was transcribed. The recombinant baculovirus has a strong infectivity to BmN cell line and to silkworm larvae and pupae. Specific hIL-11 bands were detected from all the samples of cell extract, culture supernatant, haemolymph of larvae and pupae by SDS-PAGE analysis. Biological activity of the expressed product was determined with IL-11 dependent B9-11 cell line and by MTT colorimetric assay, which indicated that biologically active rhIL-11 protein was overexpressed in BmN cell line and in silkworm larvae and pupae.
Animals ; Baculoviridae ; genetics ; Bombyx ; cytology ; growth & development ; metabolism ; Cell Culture Techniques ; Gene Expression ; Genetic Vectors ; Humans ; Interleukin-11 ; biosynthesis ; genetics ; Larva ; genetics ; metabolism ; Pupa ; genetics ; metabolism

OBJECTIVETo investigate biallelic inactivation of the von Hippel-Lindau tumor suppressor gene (VHL) in patient of renal cell carcinoma (RCC) patient.

METHODSWe extracted tumor and normal DNA from 41 RCC patients. Mutation of VHL gene from tumor tissue was detected from tumor tissue by polymerase chain reaction (PCR) and direct sequencing. Two single nucleotide polymorphism (SNP) sites located in VHL gene were analyzed by PCR restriction fragment length polymorphism, and loss of heterozygosity (LOH) was analyzed for VHL gene by comparing between tumor with normal tissue.

RESULTSMutation and LOH of VHL gene was found in 51% (21/41) and 42% (8/19) of RCC patients respectively. LOH was highly associated with mutation positive tumors (r = 0.78) and VHL biallelic inactivation was detected in 37% of RCC patients.

CONCLUSIONBiallelic inactivation of VHL gene occurs in RCC due to VHL mutation and LOH, and its frequency rate is 37%.

Adult ; Aged ; Carcinoma, Renal Cell ; genetics ; pathology ; Chromosomes, Human, Pair 3 ; genetics ; DNA Mutational Analysis ; Female ; Genes, Tumor Suppressor ; Humans ; Kidney Neoplasms ; genetics ; pathology ; Loss of Heterozygosity ; Male ; Middle Aged ; Mutation ; Polymerase Chain Reaction ; Tumor Suppressor Proteins ; genetics ; Ubiquitin-Protein Ligases ; genetics ; Von Hippel-Lindau Tumor Suppressor Protein

OBJECTIVETo prepare Form A and Form B of benazepril hydrochloride and to compare the differences in spectrums, thermodynamics and crystal structure between two polymorphic forms.

METHODSForm A and Form B of benazepril hydrochloride were characterized by Fourier transform infrared spectroscopy (IR), thermal gravimetric analysis (TG), differential scanning calorimetry (DSC), powder x-ray diffraction (PXRD) and single crystal x-ray diffraction (SCXRD).

RESULTSPreparation method, crystal structure and polymorphic stability of Form A and Form B of benazepril hydrochloride were obtained. Based on the analysis of crystal structure of both polymorphs, Form A belonged to monoclone space group P2(1) with a=7.8655(4)Å, b= 11.7700(6)Å, c= 13.5560(7)Å, β= 102.9470(10)°, V=1223.07 (11)Å(3) and Z=2, while Form B belonged to orthorhombic space group P212121, with a=7.9353(8)Å, b=11.6654(11)Å, c=26.6453(16)Å, V=2466.5(4)Å(3) and Z=4. From the DSC and XRD results, Form B of benazepril hydrochloride could be transformed into Form A after heating treatment.

CONCLUSIONForm A and Form B of benazepril hydrochloride are both anhydrous and displayed different polymorphs due to different molecular configuration. Furthermore, Form A exhibits more stable than Form B at high temperatures.

Benzazepines ; chemistry ; Crystallization ; Drug Stability ; Molecular Conformation

Objective To investigate the efficacy and clinical significance of amplification refractory mutation system (ARMS)in epidermal growth factor receptor (EGFR) gene mutation in lung adenocarcinoma.Methods Collected 566 specimens of lung adenocarcinomia in pathology from Department of the First Affiliated Hospital of Xi'an Jiaotong University from January 2015 to August 2016.As the research object,which included 34 cases of pleural cell specimens,401 cases of lung biopsy specimens and surgical specimens from 131 patients with ARMS to complete the above specimens EGFR gene mutation detection,analysis of EGFR gene mutations associated with non-small cell lung cancer patients clinical data.Results Among 566 cases of lung cancer specimens,the EGFR mutation rate of 239 cases of patients with smoking had no obvious correlation with age,gender and surgical methods(P＞0.05),but primary lung site was closely related (P＜0.05),and EGFR mutation rate of 327 cases of patients with non smoking had no obvious correlation withage,sex,operation mode and primary lung site (P＞0.05).Conclusion ARMS is an ideal method for the detection of EGFR gene mutation in non-small cell lung cancer.Smoking is a great influence on EGFR mutation rate in both lung tissue,and for right lung tissue is more dramatic.

OBJECTIVETo analyze the phenotype and genotype of two Chinese pedigrees with von Willebrand diseases, and to investigate the molecular pathogenesis.

METHODSBleeding time (BT), activated partial thromboplastin time (APTT), ristocetin-induced platelet aggregation (RIPA), von Willebrand factor-ristocetin cofactor (vWF:Rco), von Willebrand factor antigen (vWF:Ag), von Willebrand factor activity (vWF:A), von Willebrand factor collagen binding assay (vWF:CB) and multimer analysis were used for phenotype diagnosis. DNA was extracted. All of the 52 exons and exon-intron bounda ries of the VWF gene were amplified with polymerase chain reaction(PCR) and analyzed by direct sequencing.

RESULTSAPTT and BT were prolonged. Plasma RIPA, vWF:Rco, vWF:Ag, vWF:A and vWF:CB was significantly decreased. No VWF multimer can be found by plasma VWF multimer analysis. Homozygous insertional mutation g.82888_82889insCATG in exon 17 was found in proband A. Compound heterozygous mutations g.94865 G to A (Trp856stop) in exon 20 and g.110698_110699delinsG in exon 28 were found in proband B.

CONCLUSIONHomozygous insertional mutation g.82888_82889insCATG and compound heterozygous mutations g.94865G to A(Trp856X) and g.110698_110699delinsG probably have respectively induced type 3 von Willebrand diseases in the two probands.

Adolescent ; Female ; Genotype ; Humans ; Male ; Mutation ; Pedigree ; Phenotype ; von Willebrand Disease, Type 3 ; genetics ; von Willebrand Factor ; genetics

OBJECTIVETo investigate the function of abnormal fibrinogen in two inherited dysfibrinogenemia pedigrees.

METHODSRoutine coagulation tests were conducted in the probands and related family members. The antigen and activity levels of fibrinogen were detected by immunoturbidimetry assay and clauss assay, respectively. All the exons and exon-intron boundaries of the three fibrinogen genes and antithrombin gene（AT3）were analyzed by PCR amplification and direct sequencing. Routine thrombelastography (TEG) test and functional fibrinogen TEG test were both used to make a comprehensive evaluation of coagulation status and functional fibrinogen level in patients. The molecular weights of the three peptides from fibrinogen were measured by Western blot. The function of abnormal fibrinogen was assessed by fibrinogen dynamic polymerization and fibrinolysis velocity.

RESULTSThe coagulation routine tests were normal in two probands except for prolonged thrombin time (TT) and reptilase time (RT), as well as reduced activity levels of 0.5 g/L and 0.6 g/L fibrinogen, respectively. The antigen levels of fibrinogen were 2.32 g/L and 2.66 g/L in two probands, which were in the normal reference range. The genotype analysis showed that Arg275His in fibrinogen γ chain (γ Arg275His) existed in both probands and patients in these two pedigrees. Meanwhile, proband B's grandfather and aunt also carried heterozygote g.5876T>C (Ser116Pro) mutation in AT3. The results of routine TEG test demonstrated that the α values of proband B and his father were close to and lower than the lower limit of reference range, respectively, while the MA values were normal in both of them. However, functional fibrinogen TEG test revealed obviously reduced MA value. All the probands and patients demonstrated prolonged lag-off time and reduced peak value in fibrinogen dynamic polymerization tests. Meanwhile, most of fibrin formed from the patients' plasma could not be dissolved completely by plasminogen (PLG) and urokinase-typeplasminogenactivator (u-PA) at a certain time.

CONCLUSIONWe first reported cases of inherited dysgibrinogenemia associated with inherited AT deficiency. γArg275His mutation caused the abnormal fibrinogen in terms of fibrin mono polymerization and possibly in fibrinolysis. Combined use of routine TEG test and functional fibrinogen TEG test with comprehensive analyses of the parameters in both tests could better evaluate the level of functional fibrinogen and predict the risk of hemorrhage and thrombosis in patients with inherited dysfibrinogenemia.

Adult ; Afibrinogenemia ; genetics ; Child ; Female ; Fibrinogen ; genetics ; Fibrinogens, Abnormal ; genetics ; physiology ; Genotype ; Humans ; Male ; Mutation ; Pedigree ; Phenotype