Novel role of intracellular ATP in obesity pathology / 生理学报

ATP is an important energy source for cells. Traditionally, intracellular ATP levels are believed to be relatively stable and will not rise consistently in the physiological conditions. However, new studies suggest that ATP levels may rise in multiple tissues under the condition of energy surplus contributing to the metabolic disorders in obesity. However, the molecular mechanism of ATP elevation remains unknown in obesity except the increase in energy supply. Based on our experimental results and the findings reported in the literature, we discuss the cellular and molecular mechanisms by which ATP levels are regulated in cells by multiple factors, including superoxide ions, mitochondrial flash, antioxidants, anti-apoptotic molecule Bcl-xL, AMP-activated protein kinase (AMPK) and metformin. Contribution of these factors to the alteration of ATP set-point will be discussed together with their impact on insulin resistance in type 2 diabetes mellitus. With a focus on the energy surplus in obesity, we explore the mechanism of insulin resistance induced by ATP elevation and provide an answer to the contradiction between the new experimental results and the traditional viewpoint of intracellular ATP. We propose that elevation of intracellular ATP may lead to metabolic disorder in obesity through activation of a feedback mechanism that inhibits mitochondrial function.

Identification of major bioactive components and their metabolites in rat plasma after oral administration of Zhikebao tablet by UHPLC-QTOF-MS / 药学学报

An ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) method was established to identify the metabolites in rat plasma after oral administration of Zhikebao tablets. The high-resolution mass spectrometer was operated in positive and negative ion mode, respectively. First, full-scan was applied, which was dependent on a multiple mass defect filter (MMDF) combined with dynamic background subtraction (DBS). These were utilized to trigger the information dependent acquisition (IDA) function in the experiment. For the IDA criteria, the eight most intense candidate ions of per cycle were selected to do a product ion scan. Then Metabolite Pilot 2.0 software was utilized to load data to seek possible metabolites. The analytical models employed by Metabolite pilot 2.0 were established for representative compounds of the Papaveris Pericarpium and licorice in Zhikebao tablet. Finally, metabolites were identified according to accurate mass measurement and retention time. 38 components from the rat plasma after oral administration of the drug have been found, including 5 prototype opium alkaloids, liquiritin, glycyrrhizic acid and 31 relative metabolites. The metabolic transformation of Zhikebao tablet in rats was mainly induced by glucuronidation, sulfation, methylation, amine to carboxylic acid, hydrolysis and so on. In this paper, the metabolites of the main active components of Zhikebao tablet were tentatively identified, and the metabolic pathway was compared with that of single chemical drugs. Moreover, it laid the fundamental elucidation of further metabolism study of Zhikebao tablet or other compound traditional Chinese medicine preparations which containing Papaveris Pericarpium or licorice.

Electroconvulsive treatment for geriatric depression: a systematic review of randomized clinical trials / 上海交通大学学报（医学版）

Objective·To explore the clinical effectiveness and safety of electroconvulsive therapy (ECT) in elderly patients with depression.Methods·Searches were made in PubMed,EMBase,Web of Knowledge,CNKI,Wanfang,VIE CBMDisc databases and etc.CONSORT (Consolidated Standards of Reporting Trials) statement were used to assess the methodological quality of the studies.RevMan and STATA were used to do meta analysis.Results·Ten studies were included in this systematic review.The studies which used Hamilton depression rating scale (HAMD,n=5) and Montgomery-(A)sberg depression rating scale (MADRS,n=3) to access depressed symptom were analyzed.Sample size was 306 and 290 before and after treatment,respectively.The depressive symptoms were significantly improved after ECT treatment estimated by either HAMD (Z=6.39,SMD=3.64,P=0.000) or MADRS (Z=15.47,SMD=2.12,P=0.000).There was no difference of Mini-mental State Examination (MMSE) scores before and after the treatment (Z=1.47,P=0.140).The incidence of adverse events of headache,confusion,drowsiness,nausea and vomiting was more than 5％.Conclusion·Depressed symptoms are significantly improved in geriatric depression after ECT.The overall cognitive impairment is not obvious,but headache and disturbance of consciousness are common.

Identification of Imperatorin and Its Metabolites in Vitro and in Vivo by HPLC-QTrap-MS Technology / 中国药学杂志

OBJECTIVE: To identify the metabolites of imperatorin in rat urine, feces and bile after oral administration as well as the transformation products of imperatorin after incubation with rat liver microsomes with HPLC-QTrap-MS technology. METHODS: The combination use of HPLC-QTrap-MS scanning mode including multiple ion monitoring-information dependent acquisition-enhanced product ion (MRM/MIM-IDA-EPI)mode, precursor scan-enhanced resolution-information dependent acquisition-enhanced product ion (PREC-ER-IDA-EPI)mode and enhanced product ion (EPI) mode were performed for the identification of the metabolites. Based on the simultaneous appearance of [M+H]+ and [M+NH4]+ in the spectrum of PREC, the molecular weight could be unambiguously identified. The structures of compounds were then identified by the fragment ions generated from these three modes. RESULTS: With the HPLC-QTrap-MS method, 32 metabolites in urine sample, 14 metabolites in faces sample, 6 metabolites in bile sample and 17 transformation products from the rat liver microsomes sample were detected. CONCLUSION: Imperatorin is metabolized mainly in liver and excreted through kidney. The metabolic profiles of imperatorin in vivo and in vitro have good correlation.

The role of glutamate and peptidoglycan in nanotubes formation between adjacent bacteria / 中国人兽共患病学报

We investigated the formation mechanism and immediate constituents of nanotubes between adjacent bacteria,pre venting rapid increase of drug-resistant bacteria provide potential targets.We used scanning electron microscopy(SEM) to observe formation of nanotubes;glutamic was determined by surface desorption atmospheric pressure chemical ionization mass spectrometry(SDAPCI-MS)and peptidoglycan was determined by enzyme linked immunosorbent assay(ELISA) in the forma tion of nanotubes before and after.The results showed that Staphylococcus aureus and Escherichia coli consumed more glutamic after formation of nanotubes;at the same time,the concentration of peptidoglycan in nanotubes formation of S.aureus and E.coli increased significantly.This study illuminated the glutamic and peptidoglycanin role mechanism of nanotubes form.We found that peptidoglycan is constituents of nanotubes and glutamic is the main energy source for the formation of nanotubes.

Simultaneous Identification and Quantification of 9 Drugs of Opiates, Ketamine and Amphetamines in Human Plasma Using UPLC-MS/MS for Doping Control / 中国运动医学杂志

Objective To develop the method for simultaneous identification and quantification of morphine,6-monoactylmorphine,codeine,heroin,ketamine,3,4-Methylenedioxyamphetamine (MDA),3,4-methylenedioxymethamphetamine (MDMA),amphetamine and methylamphetamine in human plasma.Methods UPLC-MS/MS was adopted to analyze plasma with protein precipitated using 10％ trichloroacetic acid.Plasma samples were separated on ACQUITY UPLC HSS C18 column with aqueous solution (0.01％ formic acid)-methanol (0.01％ formic acid) as the mobile phase,and at a flow rate of 0.3 mL·min-1.The multiple reaction monitoring (MRM) mode was performed combined with the positive ion mode for quantification in four sections.Results The retention time,the characteristic fragment ions,and the relative abundance ratio of the molecular ion peak could be used to identify these nine compounds sensitively.The liner calibration curve of morphine,codeine,heroin,ketamine,6-monoacetylmorphine,MDA,MDMA,amphetamine and methamphetamine were obtained in the concentration range of 5.00×10-3～5.00 (r=0.9934),1.00× 10-2～ 10.00 (r=0.9905),1.00× 10-2～ 10.00 (r=0.9929),2.50× 10-3 ～2.50 (r=0.9960),5.00× 10-3 ～ 5.00 (r=0.9925),5.00× 10-4 ～ 5.00 (r=0.9910),5.00× 10-4 ～ 5.00 (r=0.9924),5.00× 10-4 ～ 5.00 (r=0.9920) and 5.00×10-4～5.00 μg·mL-1 (r=0.9900) respectively.The lowest detection limit was 1.00,1.00,1.00,0.50,0.50,0.10,0.10,0.10 and 0.10 ng· mL-1 respectively.The relative standard deviation (RSD) of the inter-day and intra-day were less than 18％.The relative recovery was more than 60％,and the RSD were less than 15％.Conclusion The method is accurate,sensitive and suitable for identification and quantification of 9 drugs of opiates,ketamine and amphetamines in human plasma.

The research on exprse sion of ec ll cycle and apop tosis relaet d protein in keratoaac nthoma and well-dif-ferentiated squamous cell carcinoma / 实用肿瘤学杂志

Objective To explore the expressions and significances of p 53、bcl-2、Ki-67 and apopto-sis in keratoacanthoma(KA)and well differentiated squamous cell carcinoma (WDSCC).Methods The expres-sions of p53、bcl-2、Ki-67 were detected by immunohistochemical technique in 44 cases of KA and 20 cases of WDSCC.Apoptosis was identified by using terminal deoxynucleotidyl transfers ( TdT) -mediated dUTP -biotin nick and labelling(TUNEL)method.Results The positive expressive rates of p53 in proliferative、mature and re-gressive of KA were 22.23%、26.18% and 6.52%, which was lower than the positive rate of WDSCC (41.82%).Significant differences were found between every period of KA with WDSCC in p 53 expression;The intensity of expression and pattern were similar in KA of Ki -67 and p53.There were a positive correlation be-tween p53 and Ki-67 expression rate(r=0.986,P<0.001).The bcl-2 staining showed weak expression in 1 case in WDSCC and 2 cases in KA ,but only a few positive tumor cells was limited to basal cell layer in KA .The average apoptosis rate in KA was 21.72%,which was apparently higher than in WDSCC (9.925%).There were a negative correlation between the apoptosis rate and proliferation rate of Ki -67(r=-0.824,P<0.001).Con-clusion The proliferation and apoptosis in KA can coexist .However ,the apoptosis will occupy a dominance posi-tion in regressive phase ,which results in the regression of tumor .The expressions of p53、Ki-67 and apoptosis would be a certain significance in differentiated KA and WDSCC .

Bone cement enhanced pedicle screw fixation combined with vertebroplasty for elderly patients with malignant spinal tumors / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Older patients with malignant spinal tumors are difficult to treat because they have many co-morbidities including osteoporosis. The purpose of this research is to discuss the technique and clinical outcome of bone cement enhanced pedicle screw fixation combined with vertebroplasty (the Sandwich Procedure) for elderly patients with severe osteoporosis and malignant spinal tumors.</p><p><b>METHODS</b>This study includes 28 consecutive elderly patients with malignant thoracic or lumbar spinal tumors. There were nine patients with myelomas, and 19 patients with metastatic bone tumors. The Sandwich Procedure began with curettage of the tumor and a vertebroplasty with bone cement (polymethyl methacrylate, PMMA), followed by PMMA enhanced pedicle screw fixation. Patients were evaluated with the visual analogue scale (VAS), oswestry disability index (ODI), American Spinal Cord Injury Association (ASIA) neurological function classification, and the radiographic degree of kyphosis (Cobb angle). Data were analyzed using paired t-test to compare the pre- and post-operative values. The complications, local recurrences, and the survival status were also recorded.</p><p><b>RESULTS</b>There was no operative mortality, and the mean operative time was 210 minutes (range 150 - 250 minutes). The average blood loss was 1550 ml (range 650 - 3300 ml). The average amount of cement for vertebroplasty was 3.6 ml (range 3 -5 ml). The VAS, ODI, and ASIA scores were significantly improved after surgery (P < 0.05). However, we found no differences between the pre and post-operative Cobb angles. The shortest survival time was 3 months, and we found no evidence of local recurrence in this group of patients.</p><p><b>CONCLUSION</b>The Sandwich Procedure is a safe operation and provides symptomatic relief in these difficult patients, permitting further treatment with chemotherapy or radiotherapy.</p>

Multivariate analysis of the efficacy of laparoscopic splenectomy in the treatment of immune thrombocytopenic purpura / 中华外科杂志

<p><b>OBJECTIVE</b>To identify factors that can effectively predict the efficacy of laparoscopic splenectomy (LS) in the treatment of immune thrombocytopenic purpura (ITP).</p><p><b>METHODS</b>From January 2007 to September 2012, 78 patients with ITP underwent laparoscopic splenectomy were retrospectively analyzed. According to the postoperative platelet (PLT) count and haemorrhagic manifestations, they were divided into effective group and ineffective group. Nine influencing factors were univariate analyzed and multivariate analyzed.</p><p><b>RESULTS</b>In effective group (65 cases) and ineffective group (13 cases), average PLT count of 1 day before surgery was 47×10(9)/L vs. 21×10(9)/L, average operative time was (166 ± 46) minutes vs. (139 ± 29) minutes. Univariate analysis result: PLT count of 1 day before surgery (Z = -2.776, P = 0.005) and operative time (t = 2.723, P = 0.011) was statistically significant in 2 groups, the rest factors did not significantly influence the result. Multivariate analysis revealed that only PLT count of 1 day before surgery was statistically significant (OR = 0.964, 95%CI: 0.932-0.997, P = 0.031) in 2 groups, but operative time (P = 0.051) was not statistically significant.</p><p><b>CONCLUSIONS</b>PLT count of 1 day before surgery is a predict factor in LS for ITP. Because of the limited sample number, further multi-center prospective study with large sample is warrant.</p>

Targeted blockage of RNA binding protein E2 by decoy RNA induces the granulocytic differentiation of K562 cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To use a decoy RNA targeted blockage of the RNA binding protein E2 (hnRNP E2) resulting in the CCAAT/enhancer-binding protein alpha (C/EBP alpha) gene's abnormal translation and investigate its effect on the granulocytic differentiation of K562 cells and the probable molecular mechanism.</p><p><b>METHODS</b>The hnRNP E2 decoy RNA expression plasmid was constructed and transfected into K562 cells with cationic liposome, and stable expression cells were obtained by G418 selection. The changes of C/EBP alpha and granulocyte colony-stimulating factor receptor (G-CSFR) gene expression were detected by RT-PCR and Western blot. The morphologic changes were observed after Wright-Giemsa staining. The expression of granulocytic differentiation antigens CD13 and CD15 was studied by immunocytochemistry.</p><p><b>RESULTS</b>The stably expressed pG cells were obtained. Its C/EBP alpha mRNA level remained unchanged, while 42kD-C/EBP alpha protein expression was increased by (49.7 +/- 5.5)% (P < 0.05); and G-CSFR mRNA was increased by (42.1 +/- 3.6)% (P < .05), and its protein was increased by (37.4 +/- 6.2)% (P < 0.05) compared to that in the K562 control cells. The characteristics of polymorphonuclear neutrophils appeared in pG cells and CD13 and CD15 positive cell ratios were (18.7 +/- 2.5)% and (26.3 +/- 2.9)% respectively.</p><p><b>CONCLUSIONS</b>HnRNP E2 decoy RNA can induce granulocytic differentiation of K562 cells, and G-CSF promotes this effect. The mechanisms may be that decoy RNA specifically blocks hnRNP E2, hence regulates the translation of C/ EBP alpha mRNA, restores the expression of 42kD-C/EBP alpha, and then up-regulates the expression of G-CSFR gene.</p>

Role of histamine in airway remodeling of asthmatic guinea pig / 生理学报

To investigate the role of histamine in airway remodeling, 50 healthy guinea pigs were divided into 5 groups: control group: nebulized inhalation of distilled water for 8 weeks; asthma model group: nebulized inhalation of ovalbumin (OVA) for 8 weeks after sensitization; continued asthma model group: nebulized inhalation of OVA for 14 weeks after sensitization; histamine group: nebulized inhalation of OVA for 14 weeks after sensitization and histamine was added in the last 6 weeks; antagonist group: nebulized inhalation of OVA for 14 weeks after sensitization and histamine receptor antagonists were added in the last 6 weeks. For each group, the concentration of histamine, sodium ion (Na(+)), chlorine ion (Cl(-)), arterial partial pressure of oxygen (PaO2), arterial partial pressure of carbon dioxide (PaCO2), pH, actual bicarbonate (AB), standard bicarbonate (SB) in serum, and thickness of airway mucosa, base membrane and smooth muscle were measured and compared with each other. The results showed that: (1) the concentration of histamine in serum and the thickness of airway increased, the following order was, the control group, the asthma model group, the continued asthma model group and histamine group (P<0.01); and the concentration of histamine in serum and the thickness of airway of antagonist group was lower than that of the continued asthma model group (P<0.05, 0.01). (2) PaO2 of the asthma model group was lower than that of the normal control group (P<0.01); PaO2, pH, AB, SB decreased, the following order was, the asthma model group, the continued asthma model group and the histamine group (P<0.01); and PaO2, pH, AB, SB of the antagonist group was higher than that of the continued asthma model group (P<0.01); but for PaCO2, the order was converse (P<0.01); For the concentration of Na(+) and Cl(-) in serum, there was no difference among these groups. It is concluded that: (1) Histamine is one of the mediators in the airway remodeling of asthma. (2) Histamine receptor antagonists may play a role in preventing and treating airway remodeling. (3) There is a negative correlation between the PaO2, pH and the wall thickness of the airway (P<0.01), while a positive correlation between the PaCO2, anion gap (AG) and the wall thickness of the airway (P<0.01).

Effects of histamine on endothelial nitric oxide synthase expression in pulmonary artery endothelial cells / 生理学报

All three nitric oxide synthase (NOS) isoforms are found in the lungs. It has been demonstrated that eNOS-derived NO plays an important role in modulating pulmonary vascular tone and inhibiting pulmonary vascular remodeling. Histamine induces pulmonary vasoconstriction by activating H(1)-receptor on the smooth muscle cells and vasodilation by stimulating H(2)-receptor. It remains unclear whether histamine also modulates the pulmonary vascular tone by regulating eNOS gene expression and NO production in pulmonary artery endothelial cells. Therefore, the present study was performed on cultured primary porcine pulmonary artery endothelial cells (PAECs) to investigate the effects of histamine on eNOS gene expression, and to explore the role of CaMK II in eNOS gene expression. After treatment with different concentrations histamine for different times, the levels of eNOS mRNA and protein were measured by RT-PCR and Western blot, respectively. The results showed that histamine upregulated eNOS mRNA and protein levels in a concentration- and time-dependent manner. Incubation with 10 micromol/L histamine for 24 h could increase eNOS mRNA and protein level to 160.8+/-12.2% (P<0.05) and 136.2+/-11.2% (P<0.05), respectively, of the control values. These up-regulation effects were prevented by selective CaMK II inhibitor, KN-93 (10 micromol/L). To investigate whether or not histamine increases eNOS expression by upregulating eNOS gene transcription, PAECs were transiently transfected with 1.6-kb fragment of the human eNOS promoter driving a luciferase reporter gene. The results suggested that eNOS gene promoter activity was enhanced to 148.2+/-33.7% (P<0.05) of the control after PAECs were incubated with 10 micromol/L histamine for 24 h. The nitrite and nitrate content in culture media measured by colorimetric method after incubation with 10 micromol/L histamine for 24 h indicated that the NO production in PAECs was increased. These results suggest that histamine up-regulates eNOS gene transcription and enhances NO production in PAECs by a signaling pathway involving CaMK II, which might be one of the mechanisms of histamine modulating pulmonary vascular tone.