UPLC-ESI-MS/MS-based widely targeted metabolomics reveals differences in metabolite composition among four Ganoderma species.

The Chinese name "Lingzhi" refers to Ganoderma genus, which are increasingly used in the food and medical industries. Ganoderma species are often used interchangeably since the differences in their composition are not known. To find compositional metabolite differences among Ganoderma species, we conducted a widely targeted metabolomics analysis of four commonly used edible and medicinal Ganoderma species based on ultra performance liquid chromatography-electrospray ionization-tandem mass spectrometry. Through pairwise comparisons, we identified 575-764 significant differential metabolites among the species, most of which exhibited large fold differences. We screened and analyzed the composition and functionality of the advantageous metabolites in each species. Ganoderma lingzhi advantageous metabolites were mostly related to amino acids and derivatives, as well as terpenes, G. sinense to terpenes, and G. leucocontextum and G. tsugae to nucleotides and derivatives, alkaloids, and lipids. Network pharmacological analysis showed that SRC, GAPDH, TNF, and AKT1 were the key targets of high-degree advantage metabolites among the four Ganoderma species. Analysis of Gene Ontology and Kyoto Encyclopedia of Genes and Genomes demonstrated that the advantage metabolites in the four Ganoderma species may regulate and participate in signaling pathways associated with diverse cancers, Alzheimer's disease, and diabetes. Our findings contribute to more targeted development of Ganoderma products in the food and medical industries.

Determining why continuous cropping reduces the production of the morel Morchella sextelata.

Artificial cultivation of Morchella sextelata and other morels is expanding in China, but continuous cropping reduces Morchella for unknown reasons. Here, we investigated soil that had been used or not used for M. sextelata cultivation for 0, 1, and 2 years. We found that the continuous cropping of M. sextelata substantially reduced the pH and the nutrient content of the hyphosphere soil and increased sclerotia formation by M. sextelata. Changes in the structure of bacterial and fungal communities were associated with levels of available nitrogen (N) and phosphorus in the soil. With continuous cropping, the richness and diversity of fungal and bacterial communities increased, but the abundance of Bacillus and Lactobacillus decreased and the abundance of pathogenic fungi increased. FAPROTAX analysis indicated that N cycle functions were enriched more with than without continuous cultivation, and that enrichment of N cycle and sulfate respiration functions was higher in the second than in the first year of cultivation. FunGuild analysis indicated that the functions related to pathotrophs and wood saprotrophs were enriched by M. sextelata cultivation. Overall, the results suggest that continuous cropping may reduce M. sextelata production by acidifying the soil and increasing the abundance of pathogenic fungi. Additional research is needed to determine whether increases in the abundance of pathogenic fungi and changes in soil chemistry result in the declines in production that occur with continuous M. sextelata cultivation.

Screening and verification of proteins of Salvia miltiorrhiza polyphenol oxidase interaction / 中国中药杂志

Polyphenol oxidase(PPO) is an important antioxidant enzyme in plants. It has the functions of scavenging active oxygen and synthesizing phenols, lignin, and plant protection factors, and can enhance the plant's resistance to stress and resistance to pests and diseases. Our previous research found that Salvia miltiorrhiza PPO gene can positively regulate salvianolic acid B synthesis. In order to further explore the mechanism, a pGBKT7-PPO bait vector was constructed using the cloned S. miltiorrhiza polyphenol oxidase gene(SmPPO, GenBank accession number: KF712274.1), and verified that it had no self-activation and no toxicity. The titer of S. miltiorrhiza cDNA library constructed by our laboratory was 4.75 × 107 cfu·mL~(-1), which met the requirements for library construction. Through yeast two-hybrid test, 22 proteins that could interact with SmPPO were screened. Only yeast PAL1 and TAT interacted with SmPPO through yeast co-transformation verification. Further verification was performed by bimolecular fluorescence complementary detection(BiFC). Only TAT and SmPPO interacted, so it meant that TAT and SmPPO interacted. TAT and SmPPO were truncated according to the domain, respectively. The first 126 amino acids of SmPPO and tyrosine amino transferase(TAT) were obtained to interact on the cell membrane and chloroplast. SmPPO was obtained by subcellular localization test, which was mainly loca-lized on the nucleus and cell membrane; TAT was localized on the cell membrane. Real-time quantitative PCR results showed that the SmPPO gene was mainly expressed in roots and stems; the TAT gene was expressed in roots, and the expression level in stems and flowers was low. This article lays a solid foundation for the in-depth study of the molecular mechanism of the interaction of S. miltiorrhiza SmPPO and TAT to regulate the synthesis of phenolic substances.

Screening of different AFLP fragments between near-isogenic lines of male sterile and fertile Salvia miltiorrhiza and their comparison analysis / 中国中药杂志

There is distinctive advantage of using male sterile lines to breed new cultivar and produce hybrids, when compared with general breeding method on yield and quality. In our previous work, near-isogenic lines (NILs) of male sterile and fertile Salvia miltiorrhiza have been obtained through continuous hybridization in many years. In this investigation, 378 primer combination were screened by using AFLP and BSA technique, in which 26 markers amplified from seven primers were found to tightly link to male sterile gene. Based on these markers, two linkage genetic maps were constructed. A 2 027,2 028 bp fragment was amplifed from NILs of fertile and sterile S. miltiorrhiza, respectively, using genome walking technique and previous E11/M4-208 marker as template. Four base mutations were found in intron when comparing both fragments. Among all different markers between NILs of male sterile and fertile S. miltiorrhiza, four was found to have 100% identities to chromosome 1, 3 and 5 of Arabidopsis, namely, E01/M09-418, E05/M13-308, E05/M04-750 and E01/M01-204. The E01/M09-418 marker was very close to male sterile gene of S. miltiorrhiza with distance of 2.1 cM, which also had 100% identities to male sterile gene MS2 in Arabidopsis. Both were distributed in chromosome 3 of Arabidopsis. The 2 028 bp fragment also had 100% identities to MS2 gene. Another E05/M04-750 marker that had 100% identities to chromosome 5 of Arabidopsis was found to have high identities to POP085-M05 gene of poplars and low affinity calcium antiporter CAX2 of Arabidopsis with very low E-value. The constructed genetic map and differential fragments with potential functions found in this study provide a solid foundation to lock male sterile genes in S. miltiorrhiza genome and to discover their functions.

Correlation analysis between meteorological factors, biomass, and active components of Salvia miltiorrhiza in different climatic zones / 中国中药杂志

In this study, the growth and accumulation of active components of Salvia miltiorrhiza in twenty two experimental sites which crossing through three typical climate zones. The S. miltiorrhiza seedlings with the same genotype were planted in each site in spring, which were cultivated in fields with uniform management during their growing seasons till to harvest. The diterpene ketones (dihydrotanshinone, cryptotanshinone, tanshinone I and tanshinone II(A)) in S. miltiorrhiza root samples were determined by using high-performance liquid chromatography (HPLC) method. The biomass of root (root length, number of root branches, root width and dry weight) was also measured. The results showed that tanshinone II(A) in all samples of each site were higher than the standards required by China Pharmacopoeia. It has been found there is a relationship between root shape and climate change. The correlation analysis between active components and meteorological factors showed that the accumulation of tanshinones were effected by such meteorological factors as average relative humidity from April to October > average vapor pressure from April to October > average temperature difference day and night from April to October > annual average temperature and so on. The correlation analysis between root biomass and meteorological factors exhibited that root shape and accumulation of dry matter were affected by those factors, such as average annual aboveground (0-20 cm) temperature from April to October > annual average temperature > average vapor pressure from April to October > annual active accumulated temperature > annual average temperature > average vapor pressure from April to October. The accumulation of tanshinones and biomass was increased with the decrease of latitude. At the same time, the dry matter and diameter of root decreased if altitude rises. In addition, S. miltiorrhiza required sunlight is not sophisticated, when compared with humid and temperature. To sum up, S. miltiorrhiza can adapt to a variety of climatic conditions and the southern warm humid climate is more conducive to its growth and accumulation of active components.

Citation analysis of papers from Chinese Journal of Gastrointestinal Surgery / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To analyze and evaluate the quality and academic level of the Chinese Journal of gastrointestinal surgery.</p><p><b>METHODS</b>Based on the data from WanFang data, a statistical research was carried out with the methods of bibliometrics and citation analysis to investigate the Details of the papers from Chinese Journal of gastrointestinal surgery cited by other journal articles.The important analysis indexes included number of published papers, number of cited papers, citation frequency, citation time distribution and difference, authors and citation journal distribution, etc.</p><p><b>RESULTS</b>Of all the 1442 papers published by the Chinese Journal of gastrointestinal surgery. From 2007 to 2011, 392 papers are cited, which account for about 27.18% of all cited numbers with total cited 998 times .. Immediacy index from 2009-2011 were respectively 0.08, 0.04 and 0.08. Cited half-life was 0.08 in 2011.</p><p><b>CONCLUSION</b>A relatively large proportion of papers published in the journal have ever been cited, indicating that the whole quality of the journal is considerably high. But due to its relatively short history of publication the journal still lacks papers with deep impact and great significance.</p>

P57 inhibits proliferation of U87 glioblastoma cells to reduce toxic damage induced by temozolomide / 中华神经医学杂志

Objective To analyze the relation between short-term chemotherapy of temozolomide (TMZ) and inhibition of proliferation of U87 glioblastoma cells,and investigate the role and mechanism of P57 in inhibiting U87 cells proliferation under treatment of TMZ.Methods Protein levels of P57 and proliferating cell nuclear antigen Ki-67 in U87 cells accepted TMZ short-term chemotherapy were detected by Western blotting.Protein levels of P57 in clinical primary or recurrent glioblastomas accepted TMZ short-term chemotherapy were compared by immunohistochemistry and Western blotting.After P57 was knocked down,apoptosis,cell cycle and cell vitality changes of U87 cells treated with TMZ and U87 cells with TMZ withdrawal were studied.Results After short-term chemotherapy of TMZ in U87 cells,the expression level of P57 increased but the expression level of Ki-67 decreased.The protein level of P57 in recurrent glioblastoma was higher than that in primary tumor.After P57 was knocked down in U87 cells treated with TMZ,apoptosis was enhanced and the protein level of cycle dependent kinase 2 (Cdk2) increased,which indicated that the inhibition of proliferation induced by TMZ was weakened,while the apoptosis rate increased (31.00±3.48 vs.12.83±1.40) Furthermore,after TMZ was withdrew,the cell cycle and cell vitality in U87 control cells recovered,but in U87 cells with P57 knocked down,the cell cycle was arrested and the cell vitality was reduced.Conclusion P57 inhibits cell cycle progression or proliferation of U87 cells by up-regulating P57 expression and inhibiting Cdk2 expression to reduce the damage caused by TMZ.

Comparative studies on the composition and antibiotic-resistance of pathogenic bacteria between children with community-acquired and hospital-acquired pneumonia / 中华预防医学杂志

<p><b>OBJECTIVE</b>This research was to explore the difference between children with community-acquired pneumonia (CAP) and hospital-acquired pneumonia (HAP) in the composition and antibiotic-resistance of pathogenic bacteria.</p><p><b>METHODS</b>241 CAP and 116 HAP with positive sputum culture who were hospitalized from January to December in 2008 in Children's Hospital Affiliated to Suzhou University were selected in this study. The bacteria were identified by traditionally manual method and antibiotic sensitivity tests were performed by K-B method. The chi-square or Fisher's exact test were used for statistical test.</p><p><b>RESULTS</b>In 241 CAP, Streptococcus pneumoniae and haemophilus influenza accounted for (42.2%, 106/251) and (12.4%, 31/251) infection, respectively; however in 116 HAP, Enterobacteriaceae and Non-fermenters accounted for (88.2%, 127/144). In addition, methicillin-resistant Staphylococcus aureus weren't isolated, however, its detection rate was 66.7% in HAP. The drug resistance was 1.5 times higher in HAP than that in CAP for several types of antibiotics, such as ceftazidime (37.5% (6/16) vs 75.6% (31/41)), cefepime (37.5% (6/16) vs 78.0% (32/41)), aztreonam (50.0% (8/16) vs 90.2% (37/41)), cefoperazone/sulbactam (12.5% (2/16) vs 51.2% (21/41)) and piperacillin/tazobactam (12.5% (2/16) vs 56.0% (23/41)). Klebsiella pneumoniae isolated from HAP had higher drug resistance than that isolated from CAP against some antibiotics, for example, gentamicin (0 vs 63.6% (7/11)), SMZ + TMP (20.0% (1/5) vs 63.6% (7/11)) and cefoperazone/sulbactam (0 vs 54.5% (6/11)). We also found Enterobacter cloacae isolated from HAP showed high drug resistance than that isolated from CAP against imipenem (0 vs 46.7% (7/15)), aztreonam (9.1% (1/11) vs 60.0% (9/15)) and cefoperazone (18.2% (2/11) vs 80.0% (12/15)) and Pseudomonas aeruginosa from HAP had higher resistance than that from CAP against gentamicin (0 vs 50.0% (9/18)), amikacin (0 vs 38.9% (7/18)), ceftazidime (0 vs 55.6% (10/18)), cefepime (0 vs 50.0% (9/18)) and cefoperazone (33.3% (2/6) vs 94.4% (17/18)). The detection rates of ESBLs for Escherichia coli were 84.6% (11/13) and 93.3% (14/15) in CAP and HAP, respectively (χ(2) = 0.553, P > 0.05); while for Klebsiella pneumoniae, they were 81.3% (13/16) and 95.1% (39/41), respectively (χ(2) = 2.767, P > 0.05).</p><p><b>CONCLUSION</b>CAP was mainly comprised of Streptococcus pneumoniae and haemophilus influenza; while HAP was mainly comprised of Enterobacteriaceae and Non-fermenters. The drug resistance of gram-negative bacilli was higher in HAP than that in CAP.</p>

Establishment of human glioblastoma multiform multidrug resistant cell line in vitro and identification of its biological characteristics / 中华神经医学杂志

Objective To establish the imatinib (STI-571)-resistant subline in vitro and investigate its biological characteristics. Methods Human glioblastoma multiform drug-resistant cell line (named U251AR) was established in vitro by successively increasing the concentration of imatinib in a cell culture medium. The 50％ inhibitory dose (IC50) values and the resistance indexes ([IC50U251/STI-571]/[IC50 U251]) for other chemotherapeutic agents were evaluated using cell counting kit-8 assays. Expressions of acquired multidrug resistance P-glycoprotein (MDR 1, ABCB 1; MDR3, ABCB4),breast cancer resistance protein (BCRP, ABCG2) and multidrug resistance-associated protein 1 (MRP1,ABCC1) were detected by QRT-PCR. Flow cytometry was employed to detect the protein expression of ABCG2. Results The U251AR was developed after culture for 12 months and similar morphologies of U251 and U251/STI-571 cells were determined. The resistance coefficient of U251AR cells to imatinib was 20.41 times more than that of the parent cells, and U251AR cells showed cross-resistance to many anti-tumor agents (P＜0.05). The resistance coefficients of U251AR cell line to doxorubicin and cisplatin were 5.06 and 10.28 times, respectively, more than those of U251 cells (P＜0.05). QRT-PCR indicated that the mRNA levels of MDR1, MRP1, BCRPandABCB4 (P-g4) in the U251/STI571 resistant cells were significantly higher than those in the U251 cells (P＜0.05). The protein expression of ABCG2 in U251AR cell line was significantly increased as compared with that in the parent cells (P＜0.05).Conclusion We have successfully established multidrug resistant cell line U251AR, and the drug resistance of U251/STI571 is associated with over-expressions of ABCC1, ABCB1, ABCB4, and ABCG2 mRNA, and ABCG2 protein.

Characteristics and the prevalence of respiratory viruses and the correlation with climatic factors of hospitalized children in Suzhou children's hospital / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the epidemiological characteristics of respiratory virus infection and its relations to climatic factors in Suzhou.</p><p><b>METHODS</b>From 2006 to 2009, viral etiology surveillance was conducted among 6655 children hospitalized with acute respiratory tract infections (ARIs). Direct immunofluorescence method was used to test respiratory secretion samples for respiratory syncytial virus (RSV), influenza viruses A and B (Inf-A, Inf-B), parainfluenza virus types I, II, and III (Pinf-I, Pinf-II, Pinf-III) and adenovirus. Samples were tested for human metapneumovirus (hMPV) with reverse transcription polymerase chain reaction (RT-PCR). Samples from Jan 2006 to Dec 2009 were also tested for human bocavirus (HBoV). Climatic factors, including mean temperature, relative humidity, rainfall amount, sum of sunshine and mean wind velocity were collected monthly. The relationship between activity of each virus and climatic factors were analyzed by linear regression and stepwise regression analysis.</p><p><b>RESULTS</b>From 2006 to 2009, in the total virus detection rate was 32.2% (2142/6655) in Suzhou. RSV was the most common virus and the average detection rate was 15.7% (1048/6655), followed by hMPV 8.9% (596/6655), HBoV 7.8% (148/1883), Pinf-III 2.7% (183/6655), Inf-A 2.4% (161/6655), ADV 1.3% (89/6655), Pinf-I 0.4% (29/6655), Inf-B 0.37% (25/6655) and Pinf-II 0.16% (11/6655). The positive rates of RSV, hMPV and ADV were significantly different in four years (χ(2) = 17.71, 33.23, 8.42, all P values < 0.05). Different virus has different epidemiological characteristics and distinct seasonality. The detection rate of RSV, hMPV, Inf-A were higher in Winter as 37.2%, 13.2%, 4.4%, respectively. ADV and Pinf-III were higher in summer as 2.3% and 4.6% respectively. The peak of HBoV existed in Autumn as 3.3%. The total virus detection rate showed significant inverse correlation with month average temperature (r = -0.732, P < 0.001) and a weak inverse correlation with average wind velocity was also found (r = -0.36, 0.01 < P < 0.05). The highest month total virus detection rate was from 47.6% to 84.4% when average temperature was from 3.2°C to 9.4°C and mean wind velocity was from 1.2 - 1.9 m/s. The associations of average temperature, sum of sunshine and wind velocity with RSV activity were statistical significant (r = -0.88, P < 0.001; r = -0.43, P < 0.01; r = -0.47, P < 0.01). The highest rate was from 24.3% to 58.2%, when mean temperature was from 5.3°C to 19.9°C, mean wind velocity was from 1.3 - 2.4 m/s and sum of sunshine was 61.0 to 153.4 hours. hMPV detection rate was inversely correlated with mean temperature and rain account (r = -0.43, P < 0.01; r = -0.29, P < 0.05). The rate was highest from 11.7% to 31.6% when mean temperature was from 5.3°C to 21.9°C and rain account was from 27.5 millimeter to 150.9 millimeter. Only mean temperature was positively correlated with Pinf-III (r = 0.53, P < 0.001). The rate was from 2.8% to 7.2% when mean temperature was between 11.9°C and 30.4°C. ADV detection rate was positively correlated with mean temperature and sum of sunshine, but negatively correlated with wind velocity (r = 0.35, P < 0.05; r = 0.30, P < 0.05; r = -0.32, P < 0.05). The rate was from 2.2% to 6.6% when mean temperature was between 15.9°C and 30.4°C, and sum of sunshine between 93 hours to 240.7 hours and mean wind velocity was from 1.1 - 2.8 m/s. Average temperature and relative humidity showed interactions on the detection rate of ADV (r = 0.36, P = 0.0093; r = -0.34, P = 0.016), but temperature showed higher effect on ADV detection rate. ADV detection rate was high at higher temperature (15.9 - 30.4°C) and low humidity (56% - 71%).</p><p><b>CONCLUSION</b>RSV was one of the most common viruses among hospitalized children in Suzhou, and hMPV and HBoV also played an important role in respiratory tract infection of children. Different virus has different cycle and seasonality. Climatic factors, especially mean temperature, was the main factor affecting the virus prevalence.</p>

Characteristics of human metapneumovirus respiratory tract infection in children and the relationship between the infection and meteorological conditions / 中华儿科杂志

<p><b>OBJECTIVE</b>To study the epidemiology of human metapneumovirus (hMPV) infection in children and its relations with meteorological conditions in Suzhou.</p><p><b>METHOD</b>Samples obtained from 6655 children hospitalized with acute respiratory tract infections (ARIs) during the period from 2006 to 2009, were tested for virus pathogens. Nasopharyngeal aspirates were obtained from the children according to a standard protocol and were tested for respiratory syncytial virus (RSV), influenza viruses (IFV) A and B, parainfluenza virus (PIV) types 1, 2, and 3 and adenovirus (ADV) with direct immunofluorescence assay. Samples were tested for hMPV with reverse transcription polymerase chain reaction (RT-PCR). Meteorological conditions including mean temperature, relative humidity, rainfall amount, sum of sunshine and mean wind velocity were collected monthly. The relationship between activity of the virus and meteorological conditions was analyzed by linear regression and stepwise regression analysis.</p><p><b>RESULT</b>Viral pathogens were identified in 32.2% of 6655 specimens. The positive rate of hMPV was 8.9%, RSV was 15.7%, IFV, PIV and ADV detection rates were less than that of hMPV. The annual positive rate of hMPV from 2006 to 2009 was 8.2%, 8.1%, 12.7%, 7.4% respectively (χ(2) = 33.23, P < 0.05). The hMPV positive rate of the four seasons was 11.6%, 7.6%, 4.7% and 11.7%, respectively, detection rate in winter and spring was significantly higher than those in summer and autumn (χ(2) = 74.67, P < 0.001). The positive rate of hMPV and the monthly mean temperature was moderately correlated (r = -0.43), and the monthly average rainfall (r = -0.29), monthly mean relative humidity (r = -0.27), monthly average sunshine duration (r = -0.11), the monthly average wind speed (r = -0.13) had low correlations.</p><p><b>CONCLUSION</b>hMPV was the second most common viral pathogen of acute respiratory tract infection in children in Suzhou, which prevailed predominantly in the winter and spring. Climatic factors, especially temperature and rainfall may affect the prevalence of hMPV.</p>

The prognostic value of metabolic tumor volume in FDG PET/CT evaluation of post-operative survival in patients with esophageal squamous cell cancer / 中华核医学杂志

ObjectiveTo evaluate the prognostic value of MTV on 18F-FDG PET/CT in patients with esophageal cancer.MethodsForty-nine patients with esophageal cancer underwent 18 F-FDG PET/CT scan before surgery.The median follow-up time for the patients was 29 months (range,8- 57 months).The prognostic significance of MTV,age,sex,histologic grade,SUVmax of the primary tumor,tumor size measured on PET/CT,T stage,N stage,M stage,American Joint Committee on Cancer (AJCC) stage,number and location of lymph nodes metastases were assessed by Kaplan-Meier analysis and multivariate Cox model.ResultsIn the univariate analysis,AJCC stage (x2 =16.206,hazard ratio (HR) =1.177,P ＜0.001),N stage (x2 =9.536,HR =10.833,P =0.002),T stage (x2 =5.810,HR=2.397,P=0.016),number of lymph nodes metastases (x2 =11.423,HR =1.567,P =0.001 ),and MTV (x2 =3.872,HR =2.433,P =0.049 ) were significant predictors of survival.Multivariate analysis showed that MTV and AJCC stage were independent predictors of survival (x2 =4.525,HR1.170,P =0.033 ;X2=4.875,HR =3.071,P =0.027).Kaplan-Meier survival curves revealed longer survival time of low-MTV group as compared to high-MTV group ( Log-rank,x2 =4.186,P =0.041 ).ConclusionMTV on 18 F-FDG PET/CT may be an independent prognostic factor in patients with esophageal cancer.

Eag-1 channel blocking inhibits the proliferation of glioma cells / 中华神经医学杂志

Objective To evaluate the influence of Eag-1 channel blocking on bioactivity of glioma cells in vitro. Methods Different small interfering RNAs (siRNAs) targeting for Eag-1 channel were designed and transfected to the U87 cells, and the blocking effects of those siRNAs were further confirmed on mRNA and protein levels by RT-PCR and Western blotting. The 50 nmol/l siRNAs (siRNA1 and siRNA2) and quinidine (5, 10, 20, 30 and 40 mmol/l) were used to block the activity of Eag-1 channel, respectively; and blank control group was also established. The proliferation of U87 cells 24, 48 and 72 h after the treatments was detected by MTT method; the changes of generation cycle,apoptosis ratio and intracellular reactive oxygen species (ROS) concentration were detected by flow cytometry. Results High mRNA and protein levels of Eag-1 channel on glioma cell line U87 were confirmed in the blank control group, however, siRNA1 and siRNA2 transfection groups showed significantly lower mRNA and protein levels of Eag-1 channel on glioma cell line U87. MTT method indicated that, 24, 48 and 72 h after the treatments, the proliferation of U87 cells in the siRNA1 and siRNA2 transfection groups, and quinidine treatment groups (10, 20, 30 and 40 mmol/l) was significantly inhibited as compared with that in the blank control group (P＜0.05). The IC50 value of quinidine is33.7mmol/l. As compared with the blank control group, 50 nmol/L siRNA1 and siRNA2 transfection groups, and 33.7 mmol/l quinidine treatment group enjoyed a significantly increased cell percentage at G1 stage, cell apoptosis ratio and intracellular ROS level (P＜0.05). Conclusion Eag-1 channel blocking can obviously inhibit the proliferation of glioma cells, increase the cell percentage at G1 stage and intracellular ROS level, and induce apoptosis of glioma cells.

Up-regulation of the transient A-type K+ current (IA) in the differentiation of neural stem cells of the early postnatal rat hippocampus / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Neural stem cells (NSCs) not only are essential to cell replacement therapy and transplantation in clinical settings, but also provide a unique model for the research into neurogenesis and epigenesis. However, little attention has been paid to the electrophysiological characterization of NSC development. This work aimed to identify whether the morphological neuronal differentiation process in NSCs included changes in the electrophysiological properties of transient A-type K(+) currents (I(A)).</p><p><b>METHODS</b>NSCs were isolated from early postnatal rat hippocampus and were multiplied in basic serum-free medium containing basic fibroblast growth factor. Potassium currents were investigated and compared using whole-cell patch-clamp techniques and one-way analysis of variance (ANOVA), respectively.</p><p><b>RESULTS</b>Compared with NSC-derived neurons, cloned NSCs (cNSCs) had a more positive resting membrane potential, a higher input resistance, and a lower membrane capacitance. Part of cNSCs and NSC-derived neurons possessed both delayed-rectifier K(+) currents (I(DR)) and I(A), steady-state activation of I(A) in cNSCs (half-maximal activation at (21.34 +/- 4.37) mV) occurred at a more positive voltage than in NSC-derived neurons at 1-6 days in vitro (half-maximal activation at (12.85 +/- 4.19) mV).</p><p><b>CONCLUSIONS</b>Our research revealed a developmental up-regulation of the I(A) component during differentiation of postnatal NSCs. Together with the marked developmental up-regulation of I(DR) in vitro neuronal differentiation we have previously found, the voltage-gated potassium channels may participate in neuronal maturation process.</p>

Haemopoietic stem cells from rat fetal liver in the cytothesis of radiation brain injury / 中华神经医学杂志

Objective To study the cytothesis effects of haemopoietic stem cells (HSCs) from rat fetal liver on brain injury induced by radiation. Methods Twenty-four female SD rats were randomly divided into normal control group, model group and treatment group (n=8). Total brain irradiation with 20 Gy X-ray was performed in the model group and the treatment group. Rats in the treatment group received infusion of HSCs from fetal liver (3×106 cells) 24 h, and 7, 14 and 21 d after irradiation, while rats in the model group and controls received infusion of saline at the same dosage. The serum of the rats was collected 12 h after infusion of HSCs each time; spectrophotometry was employed to detect the levels of malonaldehyde (MDA) and superoxide dismutase (SOD); the contents of tumor necrosis factor α (TNFα), interleukin (IL-1 and IL6) were measured by ELISA. Rats were sacrificed on the 28th d for assessing the pathological changes in brain tissue by HE staining and the mRNA level of Y chromosome by RT-PCR. Results Compared with that in the model group, the level of SOD in the treatment group was significantly higher, but the levels ofMDA, TNFα, IL-1 and IL6 were significantly lower in the treatment group 24 h, and 7, 14 and 21 d after the irradiation (P＜0.05). The mRNA expression of Y chromosome was noted and apoptosis of Nissl bodies was also observed in the treatment group; the cerebral cortex in the treatment group was significantly thicker as compared with that in the model group (P＜0.05); regenerative hippocampal neurons were observed in the treatment group. Conclusion HSCs from fetal liver can effectively repair the radiation brain injury.

Influence of the interruption of arteria iliaca interna distal end on penile vascularity and erectile function in male renal transplant recipients / 中华男科学杂志

<p><b>OBJECTIVE</b>It is controversial whether unilateral interruption of the arteria iliaca interna distal end affects penile hemodynamics and erectile function. The purpose of this study was to prospectively evaluate this influence by detecting the blood flow of the penile artery before and after renal transplantation.</p><p><b>METHODS</b>Thirty-three patients with chronic renal failure (CRF) on maintenance hemodialysis (MHD) received renal transplantation, the grafts revascularized by end-to-end anastomosis to the right internal iliac artery. Six months before and after the surgery, we obtained the IIEF scores of the patients, recorded their penile blood flow on color Doppler ultrasonography and the levels of serum creatinine, hemoglobin and serum cholesterol, and analyzed post-transplantation immunosuppressive medication.</p><p><b>RESULTS</b>The patients ranged in age from 21 to 55 years, of whom 36% had erectile dysfunction (ED) during MHD, and 33% after renal transplantation. A total of 67% of the renal transplant recipients (RTR) complained of unchanged and 15% deteriorated ED, while 18% admitted improved erectile function. The patients showed a significantly stronger sexual desire after the transplantation than before it (6.2 +/- 1.6 vs 8.9 +/- 0.9, P < 0.01). There was a significant decrease in peak systolic velocity (PSV) in the cavernous arteries after transplantation as compared with pre-transplantation (P < 0.01). Penile arterial blood flow insufficiency was found in none of the RTRs.</p><p><b>CONCLUSION</b>Unilateral interruption of the internal iliac artery decreases penile arterial blood flow, but not to such a degree as to result in ED. Unilateral interruption of the arteria iliaca interna distal end does not affect the erectile function of RTRs.</p>

Correlations of 99 Tc m-HL91 SPECT hypoxia imaging with HIF-1alpha and VEGF expression in non-small cell lung cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>(99)Tc(m)-HL91 ((99)Tc(m) labeled 4, 9-diaza-3, 3, 10, 10-tetramethyldodecan-2, 11-dione dioxime) is a potential noninvasive marker of tumor hypoxia. It has been reported that (99)Tc(m)-HL91 has validity for hypoxia imaging in non-small cell lung cancer (NSCLC). The aim of this study was to evaluate the (99)Tc(m)-HL91 SPECT hypoxia imaging of NSCLC, the expression of inducible hypoxia factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF), and to analyze their correlations with clinicopathological characteristics.</p><p><b>METHODS</b>Twenty NSCLC patients who underwent radical resection were enrolled into this study prospectively. (99)Tc(m)-HL91 SPECT scanning was performed in all patients at one or two days before surgery. After intravenous injection of approximately 740 MBq (99)Tc(m)-HL91, anterior, posterior and lateral planar images were collected at 2, 4 and 6 hours, respectively. Regions of interest (ROIs) were drawn in the tumor and the contralateral normal lung tissue, and the radioactivity ratio of tumor to normal tissue (T/N) was calculated. Immunohistochemistry was used to detect the expression of HIF-1alpha and VEGF in sequential histological sections of specimens.</p><p><b>RESULTS</b>Among the 20 NSCLC patients, 13 showed positive expression of HIF-1alpha and 15 had positive expression of VEGF, with a positive rate of 65.0% and 75.0%, respectively. The uptake of (99)Tc(m)-HL91 was strongly correlated with the expression status of HIF-1alpha. No correlation between HIF-1alpha and VEGF expression levels was observed. The HIF-1alpha expression level was not correlated with histological subtype, but with lymph node involvement. The expression levels of HIF-1alpha and VEGF were positively correlated with tumor stage.</p><p><b>CONCLUSION</b>The result of (99)Tc(m)-HL91 SPECT hypoxia imaging is found to be positively correlated with expression of HIF-1alpha in the non-small cell lung cancer. HIF-1alpha expression is positively correlated with VEGF expression. Furthermore, both HIF-1alpha and VEGF expressions are increasing with the increase of tumor stage.</p>

Prevention and treatment of hypertension after renal transplantation / 中国医学科学院学报

Hypertension is a common complication after renal transplantation. Among post-transplantation patients died of cardiovascular diseases, about 41% have hypertension. Hypertension is an independent risk factor for kidney transplant failure. Post-transplantation hypertension can be caused by many factors, including the use of immunosuppressants. When the blood pressure exceeds 130/90 mmHg in a kidney transplant recipient, it is reasonable to provide active medical intervention. In summary, prevention and treatment of hypertension is important to prolong the survival of kidney transplant recipients.

Appropriate use of immunosuppressants after renal transplantation / 中国医学科学院学报

Kidney transplantation has become an important method in treating advanced renal failure. Immunosuppressants play a key tool in this progress. It is important to understand the goal, mechanism, and adverse effects of immunosuppressive therapy, so as to appropriately use these drugs in post-transplantation patients on a customized basis.

Effects of PPAR-gamma agonist rosiglitazone on MMP-9 and TIMP-1 expression of monocyte-derived macrophages isolated from patients with acute coronary syndrome / 中华心血管病杂志

<p><b>OBJECTIVE</b>Coronary arterial plaque rupture and secondary thrombosis are the major pathogenesis of acute coronary syndrome (ACS). Metalloprotease (MMPs) secreted by monocyte/macrophage was the main predisposing factor of the plaque rupture and peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is involved in a variety of inflammatory cytokine gene transcriptional regulations. We explored the possible role of PPAR-gamma in the regulation of MMP-9 and TIMP-1 expressed by peripheral monocyte-derived macrophages (MDMs) from patients with ACS.</p><p><b>METHODS</b>Peripheral blood mononuclear cells were isolated from 48 patients with ACS and 28 healthy controls and stimulated by macrophage colony-stimulating factor (0.1 microg/ml for 24 hours) to form MDMs. MDMs were then incubated under various concentrations of rosiglitazone (0, 1, 10, 20 micromol/L) for 48 hours. The concentrations of MMP-9 and TIMP-1 in the supernatant were measured by enzyme linked immunosorbent assay, and the mRNA expression of PPAR-gamma, MMP-9 by RT-PCR and nuclear factor-kappaB P65 (NF-kappaB P65) expression by immunohistochemistry.</p><p><b>RESULTS</b>PPAR-gamma mRNA expression was significantly lower while NF-kappaB P65 and MMP-9 expression as well as MMP-9 and TIMP-1 concentrations in supernatant were significantly higher in ACS group than those in control group (all P < 0.05). After rosiglitazone intervention, PPAR-gamma mRNA expression was significantly upregulated in both ACS and control groups in a dose-dependent manner. Both the MMP-9 concentration in the supernatant and MMP-9 mRNA expression were reduced post intervention with rosiglitazone in both groups. The TIMP-1 mRNA expression and concentration in supernatant were not affected by rosiglitazone in both groups. Rosiglitazone induced significant downregulation of NF-kappaB P65 expression in both groups.</p><p><b>CONCLUSION</b>Rosiglitazone intervention may downregulate MMP-9 expression by upregulating PPAR-gamma expression, and by downregulating NF-kappaB expression in MDMs isolated from patients with ACS.</p>