Objective To study the inhibition of tumor necrosis factor-alpha(TNF-α)cytokine expression of BV-2 cells induced by hypoxia-reoxygenation injury by siRNA targeting TLR4 gene via the RNAi mechanisms.Method BV-2 mouse microglial cell line was cultured in six-well plates and randomly divided into group N(nor-mal group),group H(hypoxia-reoxygenation),group T(hypoxia-reoxygenation+TLR4-siRNA transfected group),group C(hypoxia-reoxygenation+pEGFP-H1/control-siRNA transfected group)and group B(hypox-ia-reoxygenation+pEGFP-H1 transfected group).Group H,group T,group C and group B were cultured in hy-poxia condition for 3 h followed by reoxygenation for 24 h.The plasma was transfected into BV-2 cells mediated by lipofectamine 2000.The efficiency of transfection were detected by flow cytometry to observe the expression of EGFP.RT-PCR method was used to detect the level of mRNA of TLR4 or NF-кB p65.Westem blot methed was used to test the expression of TLR4 protein.and ELISA was used to test the level of TNF-α in the supernatants.Analysis of variances was used for statistical analysis.Results The expression of EGFP gene waa;(67.58±7.16)% after transfection by flow cytometry analysis.Compared to group N,the TLR4 mRNA,NF-кB p65 mR-NA,TLR4 protein level and the TNF-α quantity in group H,group T,group C and group B increased after the hy-poxia-reoxygenation treatment(P＜0.01).While the expression of the TLR4 mRNA,NF-кB p65 mRNA,TLR4 protein level and the TNF-α quantity in the group T down-regulated compared to group H,group C and group B(P＜0.01).And there were no changes in group C,group B and group H about observation index(P＞0.05).Conclusions The siRNA targeting TLR4 mRNA could inhibit the inflammatory reaction released by BV-2 cells in-duced by hypoxia-reoxygenation stimulation.

A series of 6 novel 2-amino-4, 5-diarylpyrimidines were synthesized by the reaction of isoflavones with guanidine. They were identified by the spectra of IR, 1H NMR, 13C NMR and elemental analysis. The protective effects of 2-amino-4, 5-diarylpyrimidines against oxygen-glucose deprivation leading by sodium hydrosulfite in PC12 cells were studied by MTT method. It was shown that 2-amino-4, 5-diarylpyrimidines exhibited neuroprotective effects against oxygen-glucose deprivation injury in PC12 cells, which could increase the amount of survival PC12 cells and reduce their injured degree in the model.

Objective To explore the features of functional networks about recognizing dynamically positive expression with using magnetoencephalography (MEG) technology and the paradigm of expressional experiment, and further to examine its neurological basis. Methods Twelve depressed patients and twelve age,education-matched healthy controls participated to recognize the dynamically positive expression in the MEG scans. Results In comparison with the healthy,the abnormally activities regions in depressed patients during different time periods were separately as follow;decreased activation in the right inferior parietal lobule(t = 3.94),the right su-pramarginal gyms(t = 3.59),the bilateral posterior cingulated(t = 5. 04, t = 4. 85) ,the bilateral precuneus(t = 3.84) ,the left cuneus(t = 3.58) and increased activation in the bilateral hippocampus(t = 2. 13, t = 2.56) ,the left amygdale(t = 2.41), the bilateral uncus gyrus(t = 2.33, t = 2.44) , the right anterior cingulated (t = 2. 15) , the left fusiform gyms (t = 3. 33) (P < 0. 05, unconnected). Conclusion The results indicate, compared with healthy controls,abnormal brain activities as the weakened function of the medial temporal cortex,parietal lobe and the enhanced function of limbic system were distributed extensively in depressed patients during recognizing dynamically positive expression. These abnormalities may prompt the dynamic characteristics of the magnetic source imaging about recognizing dynamically positive expression,and further reveals the mechanisms of emotional symptoms in depression.

Objective To determine whether it is necessary to execute pre-labor pubic shaving.Methods Women(336 cases)who received pubic shaving through vaginal delivery from 2003 to 2006 were set as the control group.Women(103 cases)who were waiting for delivery currently in our hospital were set as the test group.The test group only received perineal cleaning without pubic shaving.If episiotomy was needed during the operation,the perineal hair Was cut by scissors.The perineum infectious rate and postpartal infection Was compared in the two groups.We interviewed the patients attitude for pubic shaving and counted the numing time used for pubic shaving.Results The perineal infection in the test group was 5.8%,which was no statistically different from that of the control group(2.4%),U=0.33,P＞0.05.Totally 95% women in the test group support this method demonstrated humanization.In the telephone folow-up visit for patients in the control group they comphined itch and discomfort during the regeneration of pubes.The average numing time for pubic shaving Was(4.6±0.6)minutes.Conclusions No pubic shaving for pre-labor skin preparation was feasible.If operation was disturbed by pubes we could use scissors to cut it short.This method could reduce the skin infection and propagation of blood-derived infectious disease.It increased the comfort of parturient,saved time and proved to be a nursing method of humanization.

Objective To explore the gender differences of brain activation between patients with depression in recognition of dynamically sad expression with magnetoencephalography (MEG) technology and the paradigm of expressional experiment,and further to examine its neurological basis. Methods Brain activities were detected using MEG during the recognition of dynamic facial video with 12 female depressed patients, 11 male depressed patients, and their corresponding normal controls matched by age and education. Results Compared with male depression,female depression showed increased activation during different time periods were in turn as follows: bilateral inferior frontal gyrus ( x, y, z = 58/- 60,12/22,20/20, T = 2.17,1.88 ), left anterior cingulate gyrus ( x, y, z = 64,12,4, T = 2.37 ), bilateral superior frontal gyrus ( x, y, z= 30/-24,64/- 4,0/50, T = 2.86,2.36 ),the left brainstem( x, y,z = - 12, - 16, - 18 ,T = 1.96) ,right middle frontal gyrus( x,y,z =40,60,4,T = 2.22 ),bilateral precentral gyrus(x,y,z = -62/56, 16/10, 10/8. T = 1.99,2.65 ) ,left hippocampus ( x,y,z = - 26,- 14, -20,T = 1.77) ,left parahippocampal gyrus(x,y,z = - 16, - 12, -20,T = 1.72) ,right middle occipital gyrus( x ,y ,z = 50, -76, -14 ,T = 1. 75 ),while the decreased brain activation were in turn in right superior temporal gyrus ( x, y, z = 68, - 18,2, T = 1.85 ), right parahippocampal gyrus ( x, y, z = 20, - 36, - 14, T = 1.92 ), left cuneus( x,y,z = -6, -98,14,T =2.46) ,bilateral temporal middle gyrus( x,y,z =52/-34, - 80/-62,20/28,T = 1.95,2.57 ), left pre-cuneus ( x, y, z = - 40, - 74,38, T = 2.58 ), right hippocampus( x, y, z = 24, - 32, - 18,T = 2.05 ) in recognition of dynamically sad expression. ( p ＜ 0. 05, corrected). Conclusion The data suggest that there may be significant gender differences of functional network in depressed patients during recognizing dynamical sad facial expression.

A sesquiterpene synthase (AsSS4) full-length open reading frame (ORF) cDNA was cloned from wounded stems of Aquilaria sinensis by RT-PCR method. The result showed that the ORF of AsSS4 was 1,698 bp encoding 565 amino acids. Prokaryotic expression vector pET28a-AsSS4 was constructed and transformed into E. coli BL21 (DE3) pLysS. Recombinant AsSS4 protein was obtained after induction by IPTG and SDS-PAGE analysis with a MW of 64 kD. Enzymatic reactions using farnesyl pyrophosphate showed that recombinant AsSS4 protein purified by Ni-agarose gel yielded five sesquiterpene compounds, cyclohexane, 1-ethenyl-1-methyl-2, 4-bis(1-methylethenyl)-, β-elemene, α-guaiene, α-caryophyllene and δ-guaiene. This paper reported the first cloning and functional characterization of AsSS4 gene from A. sinensis, which will establish a foundation for future studies on the molecular mechanisms of wound-induce agarwood formation in A. sinensis
Alkyl and Aryl Transferases ; biosynthesis ; genetics ; Azulenes ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; Open Reading Frames ; Polyisoprenyl Phosphates ; Recombinant Proteins ; biosynthesis ; Sesquiterpenes ; metabolism ; Sesquiterpenes, Guaiane ; Thymelaeaceae ; enzymology ; genetics

Objective To explore the difference in the intensity of brain activation during the recognition of sad facial expression and changes in neuropsychology cognition in patients with a first depressive episode and with recurrent depression.Methods 11 patients with a first major depressive episode,11 patients with recurrent major depression and 11 healthy controls matched in all aspects were scanned to compare the intensity of brain activation in the process of sad and neural facial expression recognition Results Compared with the healthy,first-episode depressed patients showed increased activation in right cuneus(BA18),right fusiform gyrus(BA20),left medial frontal gyrus(BA6),left middle temporal gyrus(BA39),left parahippocampal gyrus(BA30)and decreased activation in right temporal gyrus(BA22),right parahippocampal gyrus(BA27);recurrent depressed patients showed increased activation in right medial frontal gyrus(BA6),right superior temporal gyrus(BA38)but no activation increased.In comparation with the first-episode depressed,the recurrent,showed decreased activation in right middle frontal gyrus(BA6),fight thalamus,right superior temporal gyrus(BA22),right precentral gyras(BA44),right claustrum and increased activation in right parahippocampal gyrus(BA28),right claustrum,right insula(BA13). Conclusion The neural correlates of patients with depression,especially the first-episode depressed for recognition of sad facial expression is different from the healthy with more brain regions involved in the process.There are also differences in the neural basis for recognition of sad facial expression between patients with a first depressive episode and patients with recurrent depression.Performances of brain regions related with emotional recognition de-clined more in the recurrent depressed.

Objective To investigate the effects of total anthraquinone in rheum on aquaporin 2 and aquaporin 4 expression in rat kidney and explore its diuresis mechanism.Methods Thirty-two SD rats were randomly divided into control group,low dose group,medium dose group and high dose group.Total anthraquinone in rheum was administered to rats at different doses.Urinary volume of 24 h,Na+ concentration and osmolality were detected.Rats were sacrificed 5 days later.Blood samples were taken from the abdominal aorta to detect blood biochemical indicators. Kidneys of rats were removed to detect AQP2, AQP4 expression through immunohistochemistry,Western blot,and RT-PCR. Results Compared with control group,there were significantly increased 24 h urine output of rats in medium and high dose group[(16.21±1.96),(18.16±1.8) ml vs(13.85±1.25)ml,P＜0.05].24 h urine output in low-dose group did not change significantly.AQP2 protein and mRNA expression were significantly decreased in rats'kidneys of medium and high dose group (P＜0.01),The AQP4 protein and mRNA expression was significantly down-regulated in high dose group (P＜0.01).In medium does group,the AQP4 protein expression was down-regulated (P＜0.01),without significant decrease in the mRNA expression.Protein and mRNA expression of AQP2 and AQP4 did not significantly change in low dose group.Conclusion Total anthraquinone in rheum can reduce the expression level of AQP2 and AQP4 in rat kidney,which is probably one mechanism of diuresis caused by rheum.

Objective To investigate the performance of left ventricular torsion (LVtor) or twist with cardiac dysfunction patients by speckle tracking imaging (STI). Methods Standard left ventricular short-axis views images were acquired by rountine two-dimensional echocardiography in thirty-two healthy humans and twenty-four patients with cardiac dysfunction. LVtor angle,left ventricular rotation (LVrot) angle,the peak value and time of LVtor and LVrot were measured respectively. The data were compared and analyzed between the two groups. Results Compared with normal group,the peak of LVtor and LVrot angle was decreased in cardiac dysfunction group at both apical and basal planes (P<0.05). The peak time of LVtor in patients with cardiac dysfunction was delayed (P<0.05). The peak time of clockwise rotation at the apical level segments was longer than control group (P<0.05). Conclusions There is a temporal sequence difference of left ventricular rotation between basal and apical planes during left ventricular contraction and decreased LVtor has been demonstrated to influence left ventricular function.

OBJECTIVETo investigate the correlation between mutation in EGFR tyrosine kinase domain and tumor response as well as prognosis in advanced stage non-small cell lung cancer (NSCLC) treated with iressa.

METHODSFrom May 2002 to Feb. 2005, iressa was orally administered at a dose of 250 mg once daily for 106 advanced stage NSCLC patients until occurrence of disease progression or intolerable toxicity. Cancer tissue was obtained from these patients, and DNA was extracted for analysis of mutation in exon 18 to 24 of EGFR. Exon 18 to 24 of EGFR were amplified by nest PCR, sequenced and analyzed from both sense and antisence directions.

RESULTSPrimary NSCLC tissue specimens consisted of 25 frozen tissue blocks and 81 paraffin-embedded tumor tissue blocks from 106 consecutive NSCLC patients. Mutation was found to be more frequent in the adenocarcinoma than in the squamous cell carcinoma (35.9% vs 14.3%, P =0.033). Mutation was identified in 32 patients (30.2%). Response rate to iressa was 71.9% in the patients with EGFR mutation versus 13.5% in those without mutation (P <0.01). Compared with the patients without EGFR mutation, those with mutation had longer overall survival (median, 13.45 vs. 5.25 months; P<0.01) and median time to progression (median, 8.35 vs. 3.0 months; P <0.01).

CONCLUSIONEGFR mutation may be positively correlated with the response and survival in advanced stage Chinese NSCLC patient treated with iressa.

Adenocarcinoma ; drug therapy ; genetics ; pathology ; Adolescent ; Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; genetics ; pathology ; Carcinoma, Squamous Cell ; drug therapy ; genetics ; pathology ; Exons ; Female ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Lung Neoplasms ; drug therapy ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Point Mutation ; Prognosis ; Quinazolines ; therapeutic use ; Receptor, Epidermal Growth Factor ; genetics ; Sequence Deletion