A solution with different Cu supply levels was cultured to investigate gama-aminobutyric acid (GABA) accumulation in Elsholtzia splendens, a native Chinese Cu-tolerant and accumulating plant species. Increasing Cu from 0.25 to 500 micromol/L significantly enhanced levels of GABA and histidine (His), but considerably decreased levels of aspartate (Asp) and glutamate (Glu) in the leaves. The leaf Asp level negatively correlated with leaf Cu level, while leaf GABA level positively correlated with leaf Cu level. The leaf Glu level negatively correlated with leaf GABA level in Elsholtzia splendens. The depletion of leaf Glu may be related to the enhanced synthesis of leaf GABA under Cu stress.
Copper ; toxicity ; Dose-Response Relationship, Drug ; Drug Tolerance ; physiology ; Gene Expression Regulation ; drug effects ; Lamiaceae ; drug effects ; metabolism ; Plant Leaves ; drug effects ; metabolism ; gamma-Aminobutyric Acid ; metabolism

Objective To compare the prophylactic efficacy of combination of ganciclovir and acy- clovir or acyclovir alone against cytomegalovirus pneumonia in renal transplant recipients.Methods A to- tal of 217 renal transplant recipient(124 men and 93 women;mean age,32 years;age range,16-72 years) were divided into 3 groups randomly.In 51 cases,acyclovir was taken orally at a dose of 400 mg,3/d,from the third d to 3 months after transplantation.In 74 cases,ganciclovir was administered at a dose of 250 mg/d intravenously from the 21st d to 27th d to replace Acyclovir.In 92 cases,no prophylaxis against eytomegalov- irus pneumonia was performed.All patients were followed 3 months after transplantation.Comparison of the incidence rates of cytomegalovirus pneumonia among the 3 groups was performed using Fisher's exact test. Results Cytomegalovirus pneumonia developed in 20 cases in the 3 groups,including 4 cases(5.4%) in combined use group,2 cases(3.9%)in acyclovir alone group,and 14 cases(15.2%)in control group. Significant difference existed between the 2 experimental and control groups(P＜0.05).However,no signifi- cant difference existed between the 2 experimental groups(P＞0.05).Of the 20 cases,17(85.0%)were cured,and 3 died of respiratory failure.Conclusions Ganciclovir and acyclovir have prophylactic effect a- gainst cytomegalovirus pneumonia in renal transplant recipients.These 2 medications are inexpensive,and the patients have good compliance.

OBJECTIVETo study the expression of NFkappaB p65 and its target genes in intestinal metaplasia (IM), dysplasia (Dys), gastric cancer (GC) infected with Helicobacter pylori (Hp) and explore the mechanism of infection by cytotoxin-associated antigen A expressing Hp (CagA(+)Hp) in the development of gastric cancer.

METHODSCagA antibody in blood sample of 289 patients was determined by ELISA. Hp was detected by rapid urease test and Warthin starry staining. Expression of NFkappaB p65 and its target genes in IM, Dys and GC was examined by immunohistochemistry.

RESULTSIn IMI approximately II, IMIII, DysI, DysII approximately III and GC, the expression of NFkappaB p65 was significantly higher in patients with CagA(+)Hp infection than those without CagA Hp infection. In IMIII and DysII approximately III, the expression of NFkappaB p65, c-myc, CyclinD(1) and bcl-xl was significantly higher in patients with CagA Hp infection than those without CagA Hp infection. In gastric cancer infected with CagA(+)Hp, the expression of NFkappaB p65, c-myc, CyclinD(1) and bcl-xl was significantly higher in intestinal type than in diffuse type.

CONCLUSIONThere are different mechanisms in intestinal type and diffuse type in the development of gastric cancer. The occurrence of intestinal type gastric cancer is associated with CagA(+)Hp infection which by NFkappaB p65 upregulating the expression of c-myc, CyclinD(1),bcl-xl in patients with IMIII, DysII approximately III. It may be an effective method to prevent gastric cancer by inhibiting NFkappaB p65.

Adult ; Aged ; Antigens, Bacterial ; analysis ; Bacterial Proteins ; analysis ; Cyclin D1 ; metabolism ; Female ; Helicobacter Infections ; complications ; metabolism ; microbiology ; Helicobacter pylori ; Humans ; Male ; Middle Aged ; Precancerous Conditions ; metabolism ; microbiology ; pathology ; Proto-Oncogene Proteins c-myc ; metabolism ; Stomach Neoplasms ; metabolism ; microbiology ; pathology ; Transcription Factor RelA ; genetics ; metabolism ; bcl-X Protein ; metabolism

The goal of this study is to research the genetic characteristics and relationship between HN and P genes of NDV. The nucleotide sequence and deduced amino acid sequence were analyzed for the Hemagglutinin-neuramindase (HN) and Phosphoprotein (P) gene of twelve field isolates of Newcastle disease virus (NDV) during 1997-2005 in China. The HN and P gene sequences of fifteen NDV reference strains from GenBank were also used in this study. The molecular evolution distance of nucleotides and amino acids were calculated by MEGA 4.0 software, and analysis of variance and correlations were analyzed by SPSS11.0 software among different length sequences of the HN gene or P gene. The nucleotide and amino acids correlation of HN and P gene were analyzed respectively. The correlation of evolution distance and isolation year were also calculated. The results indicated that there were difference and good correlation of nucleotide and amino acid among different length sequences of the HN gene or P gene. These results revealed that the HN and P gene of NDV have the different response to selective pressure to adopt to landscape and closely relationship on heredity mutations. Nucleotide variations of HN and P gene have relationship with isolation year of strains.
Evolution, Molecular ; Genetic Variation ; HN Protein ; genetics ; Newcastle disease virus ; classification ; genetics ; Phosphoproteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the influence of enteral administration of carbachol on the intestinal dysfunction of both severely burn patients and rabbits with partial intestinal ischemia/reperfusion (I/R) injury.

METHODSSeventy-five white rabbits were inflicted with I/R injury and randomized into intestinal I/R (I, n=25), carbachol [C, n=25, with 3g/L carbachol (3 mg/kg) injection into duodenum 1 h after SMA occlusion] and sham operation (SO, n=25, with SMA isolation but no occlusion) groups, and 5 other as normal controls. The blood flow of intestinal mucosa was detected before and after SMA occlusion or admission of carbachol. Changes in diamine oxidase (DAO), D-lactate, xylopyranose absorption, blue dextran discharging time were measured at 2, 4, 6, 8, 24, 48, 72 h after SMA occlusion. In addition, eight severe burn patients with TBSA of 84 +/- 12% were enrolled in the study, and carbachol (15 microg/kg) was administered to patients when abdominal distension or bowel sound was lower than 2 times/min, then the number of abdominal distension and bowel sounds per minute were observed.

RESULTSThe blood flow in intestinal mucosa of rabbits without SMA occlusion was (102 +/- 5) PU, reduced to (48 +/- 6) PU after SMA occlusion, and increased to (77 +/- 3) PU after injection of carbachol. The plasma DAO activity and D-lactic acid content in I group began to increase 4 hours after SMA occlusion, and they reached the peak 24 hours after SMA occlusion (4.63 +/- 0.27 U/ml, 7.9 +/- 2.4 mg/L) , after that they decreased gradually, but still higher than the normal value (0.89 +/- 0.14 U/ml, 2.0 +/- 1.1 mg/L, P < 0.05). In carbachol group, data showed the same trends as that in intestine I/R group with lower values, while no obvious changes were in sham operation group (P > 0.05). The content of D-lactic decreased dramatically 2 hours after D-lactic administration in both I and C groups, increased 6 hours after SMA occlusion, then decreased gradually, but it in C group was always higher than normal values, and little fluctuation was in sham operation group. There was no blue dextran discharge 2 hours after SMA occlusion. The discharging distance increased 6 hours later, but it was obviously shorter than the normal value 24 hrs after operation (P < 0.05) , then it returned to normal 48 to 72 hrs after operation. In the C group, blue dextran discharge was found immediately after its injection, with obvious increase in the discharging distance to peak value (43 +/- 6 cm) 6 hours after injury, and returning to normal (28 +/- 3 cm) gradually. In severe burned patients, the bowel sounds was (1.6 +/- 1.1) per minutes before carbachol administration, then increased dramatically to (6.9 +/- 1.7) per minutes 10 mins after administration, reached to a higher level 30 minutes after administration (8.3 +/- 2.4 ) times/min, and it maintained to (6.1 +/- 1.3) times/min 1 hour after administration. Abdominal distension was ameliorated 2 hours after carbachol administration, six patients were able to defecate.

CONCLUSIONEnteral administration of Carbachol can increase the blood flow of intestine mucosa, help to improve the movement, absorption and barrier functions of intestine, and ameliorate intestinal dysfunction in patients with severe burns.

Adolescent ; Adult ; Animals ; Burns ; drug therapy ; physiopathology ; Carbachol ; therapeutic use ; Disease Models, Animal ; Female ; Humans ; Intestinal Mucosa ; blood supply ; metabolism ; Intestines ; physiopathology ; Male ; Rabbits ; Reperfusion Injury ; drug therapy ; physiopathology

Objective: To explore the expression and prognostic significance of miR-223 in patients with mantle cell lymphoma (MCL) and to investigate the possible mechanism. Methods: Twenty-one newly diagnosed MCL patients with bone marrow involvement were enrolled in the present study, 20 healthy donors as normal control. The expression level of miR-223 and SOX11 mRNA was determined by RQ-PCR. CCK-8 and flow cytometer assays were used to analyze cell proliferation, cell cycle and apoptosis of the constructed miR-223 overexpressing MCL cell line, Granta519 cells. SOX11 protein expression level was determined by Western blot. The target gene of miR-223 was confirmed by dual luciferase reporter assay. Results: ①Of the 21 newly diagnosed MCL patients, 15 were male and 6 female, the median age was 58 (37-72) years. The expression level of miR-223 was significantly down regulated in MCL patients compared with that of healthy donors (14.7±10.5 vs 1 244.1±1 935.2, P<0.001). The lower expression of miR-223 was inversely correlated with high-risk mantle international prognostic index (P=0.001), elevated LDH (P=0.001), ECOG score ≥2 (P=0.035). ②Using the median relative expression level of miR-223 as the cutoff value, 21 MCL patients were divided into high-expression group (n=10) and low-expression group (n=11) and found that the high-expression group had a significantly superior OS (median OS: 36 vs 12 months, P=0.021). ③In vitro results showed that compared with the control group, the proliferation of miR-223 overexpressed Granta519 cells was inhibited (the most significant reduction on 96h, P<0.001), manifested by lower proportion of cells in G2/M phase (P<0.001) and increased apoptosis (P<0.001), and the expression level of SOX11 protein in Granta519 cells was significantly lower than that of the control group. ④miR-223 could inhibited the 3' untranslated region of SOX11, and the expression level of miR-223 was significantly negatively correlated with mRNA level of SOX11 in MCL patients (r=-0.81, P<0.001). Conclusions: The expression of miR-223 was repressed in MCL and was associated with poor clinical outcomes, which may be probably attributed to its direct targeting SOX11.
Adult ; Aged ; Female ; Humans ; Lymphoma, Mantle-Cell ; Male ; MicroRNAs ; Middle Aged ; Prognosis ; RNA, Messenger ; SOXC Transcription Factors

Elsholtzia argyi and Elsholtzia splendens, which are Chinese endemic Pb/Zn mined and Cu mined ecotype respectively, were investigated in the aspect of their response to Pb toxicity in the presence or absence of EDTA addition. After 8 d's Pb treatment, root length, root surface area and root volume of E. splendens decreased much more than those of E. argyi, and reduced considerably with increase of Pb, while no marked change was noted for root average diameter. Compared to E. argyi, length of root with diameter (D)<0.2 mm was significantly reduced for E. splendens as Pb increasing. Root with cross-sectional area of D<0.1 mm for E. splendens was at Pb> or =10 mg/L, while for E. argyi, it was at Pb> or =25 mg/L. DW of E. splendens decreased much more than that of E. argyi with increase of Pb. E. argyi exhibited much more tolerance to Pb toxicity than E. splendens. Treatment with 100 mg/L Pb plus 50 mmol/L EDTA significantly decreased the length and surface area of D< or =0.2 mm root, increased the length and surface area of 0.2< or =D< or =0.8 mm root for the case of E. argyi, while for E. splendens, length and surface area of D<0.6 mm root reduced, as compared to 100 mg/L Pb treatment, alone. At 100 mg/L Pb, shoot Pb accumulation in E. splendens and E. argyi were 27.9 and 89.0 microg/plant DW respectively, and much more Pb was uptaken by the root and translocated to the stem of E. argyi as compared to E. splendens. Treatment of the plant with 100 mg/L Pb plus 50 mmol/L EDTA increase leaf Pb accumulation from 16.8 to 84.9 g/plant for E.splendens and from 18.8 to 52.5 g/plant for E. argyi, while both root and stem Pb pronouncedly reduced for both Elsholtzia species. The increased translocation of Pb to the leaf of E. splendens than that of E. argyi at the treatment of 100 mg/L Pb plus 50 mmol/L EDTA should be further investigated.
Biodegradation, Environmental ; Cell Proliferation ; Dose-Response Relationship, Drug ; Industrial Waste ; prevention & control ; Lamiaceae ; anatomy & histology ; classification ; drug effects ; metabolism ; Lead ; pharmacokinetics ; toxicity ; Plant Roots ; anatomy & histology ; drug effects ; metabolism ; Species Specificity

Copper accumulation and intracellular distribution in Elsholtzia splendens, a native Chinese Cu-tolerant and accumulating plant species, was investigated by transmission electron microscope (TEM) and gradient centrifugation techniques. Copper concentrations in roots, stems and leaves of E. splendens increased with increasing Cu levels in solution. After exposure to 500 micromol/L Cu for 8 d, about 1000 mg/kg Cu were accumulated in the stem and 250 mg/kg Cu in the leaf of E. splendens. At 50 micromol/L Cu, no significant toxicity was observed in the chloroplast and mitochondrion within its leaf cells, but separation appeared at the cytoplasm and the cell wall within the root cells. At >250 micromol/L Cu, both root and leaf cell organelles in E. splendens were damaged heavily by excessive Cu in vivo. Copper subcellular localization in the plant leaf after 8 days' exposure to 500 micromol/L Cu using gradient centrifugation techniques was found to be decreased in the order: chloroplast>cell wall>soluble fraction>other organelles. The plant root cell wall was found to be the site of highest Cu localization. Increase of Cu exposure time from 8 d to 16 d, increased slightly Cu concentration in cell wall fraction in roots and leaves, while that in the chloroplast fraction decreased in leaves of the plants grown in both 0.25 micromol/L and 500 micromol/L Cu. TEM confirmed that much more Cu localized in cell walls of E. splendens roots and leaves, but also more Cu localized in E. splendens' chloroplast when the plant is exposed to Cu levels>250 micromol/L, as compared to those in the plant grown in 0.25 micromol/L Cu. Copper treatment at levels>250 micromol/L caused pronounced damage in the leaf chloroplast and root organelles. Copper localization in cell walls and chloroplasts could mainly account for the high detoxification of Cu in E. splendens.
Copper ; analysis ; metabolism ; Lamiaceae ; cytology ; metabolism ; ultrastructure ; Microscopy, Electron, Transmission ; Plant Leaves ; cytology ; metabolism ; ultrastructure ; Plant Roots ; cytology ; metabolism ; ultrastructure ; Plant Stems ; cytology ; metabolism ; ultrastructure

Objective To investigate the changes in the tight junctions of the blood-brain barrier (BBB) in a rat model bearing C6 glioma. Methods Rat models bearing C6 glioma were established in 30 male Wistar rats by stereotactic injection of C6 glioma cells into the caudate nuclei, with another 30 rats as the normal control group. Twenty-one days after the injection, magnetic resonance imaging (MRI) was performed, and the normal brain tissue, tumor core, tumor margin and the ipsilateral hemisphere tissues 2 mm from the tumor margin were sampled for ultrastructural observation of the BBB using electron microscopy, Immunohistochcmistry and RT-PCR were used to analyze the expression of claudin-5 protein and mRNA in these tissues, respectively. Results MRI revealed tumor formation in the brain 21 days after C6 cell injection. In normal control brain tissues, the paracellular cleft between the adjacent endothelial cells was sealed by continuous tight junction strands, which were found in only 22.23% of the microvesscls in the core of the brain gliomas, and obvious paracellular clefs were found between the adjacent endothelial cells in other microvessels. In the tissues on the tumor margin, intercellular tight junctions were found in 57.15% of the microvessels with the rest microvesseis having obvious paracellular clefts. Immunohistochemistry showed strong claudin-5 positivity in the control brain tissue but yielded negative results in the core of the gliomas. Compared with the core of the gliomas, the tissues on the tumor margin, those 2 mm from the tumor margin and the control brain tissues showed significantly increased claudin-5 expression (P<0.05,respectively). Conclusion In C6 glioma-bearing rats, the continuity of the tight junctions in the BBB is interrupted due to decreased expression of claudin-5 in the brain gliomas.

In order to investigate the transmission risk of H5N1 avian influenza viruses (AIV) from sewage in Changsha poultry markets, the evolution relationship and molecular characteristics of non-structural (NS) genes of H5N1 AIV from sewage were analyzed. Nine H5N1 AIV environmental sewage specimens were collected from Changsha poultry markets. The NS genes were amplifyed by PCR and then sequenced with TA cloning. Amino acid(aa) sequence alignment and phylogenetic tree analysis were conducted by Lasergene and Mega5 software. Eight NS genes TA cloning were constructed successfully. Phylogenetic tree indicated that they were belonged to the allele A subgroup. Aa homology analysis showed 90.1% 92.5% identity in NS1 proteins and 91.0% - 92.6% identity in NS2 proteins compared with reference viruses of the allele A (A/chicken/ Hubei/ w h/ 1999). The homologies of the amino sequences of NS1 and NS2 in this study were 93.8%-100.0% and 98.4%-100.0%, respectively. The C terminal of all eight H5N1 NS1 proteins from sewage in poultry markets carried a ESEV of PL motif and the 92 amino acids were E, furthermore, the 80 to 84aa were missed which were the characteristics of highly pathogenic AIV. The NS genes of H5N1 AIV from sewage in poultry markets have molecular characteristics of highly pathogenic and have the potential risk of H5N1 virus spreading.
Amino Acid Sequence ; Animals ; Influenza A Virus, H5N1 Subtype ; chemistry ; classification ; genetics ; isolation & purification ; Influenza in Birds ; transmission ; virology ; Molecular Sequence Data ; Phylogeny ; Poultry ; Sequence Homology, Amino Acid ; Sewage ; virology ; Viral Nonstructural Proteins ; chemistry ; genetics