Adenocarcinoma ; blood supply ; metabolism ; mortality ; secondary ; Adult ; Aged ; Female ; Humans ; Lymphatic Metastasis ; Male ; Microcirculation ; Middle Aged ; Neovascularization, Pathologic ; metabolism ; pathology ; Prognosis ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Cell Surface ; biosynthesis ; genetics ; Receptors, Urokinase Plasminogen Activator ; Stomach Neoplasms ; blood supply ; metabolism ; mortality ; pathology ; Survival Rate ; Vascular Endothelial Growth Factor A ; metabolism

To clone the 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase (TwMCT) full length cDNA from Tripterygium wilfordii, the specific primers were designed according to the transcriptome data and the LCPCR were carried out. After a series of bioinformatics analysis on the TwMCT, the MeJA induced expression content were investigated by real-time fluorescence quantification polymerase chain reaction (RT-qPCR). The result showed that the full of TwMCTcDNA was 1 318 bp nucleotides encoding 311 amino acids. The molecular weight of the deduced TwMCT protein was about 34.14 kDa and the theoretical isoelectric point was 8.65. Result of the RT-qPCR analysis indicated that the content of TwMCT mRNA expression in T. wilfordii suspension cell was rising after treating with MeJA and reached the maximum in 24 h. Cloning and analyzing TwMCT gene from T. wilfordii provided gene element for studying the function and expression regulation of secondary metabolites.
Amino Acid Sequence ; Cloning, Molecular ; Erythritol ; analogs & derivatives ; metabolism ; Gene Expression Regulation, Plant ; Molecular Sequence Data ; Nucleotidyltransferases ; chemistry ; genetics ; metabolism ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Protein Structure, Secondary ; Sequence Alignment ; Sugar Phosphates ; metabolism ; Tripterygium ; chemistry ; enzymology ; genetics

OBJECTIVETo investigate relationship between expressions of hypoxia inducible factor-1alpha (HIF-1alpha) and insulin-like growth factor-II (IGF-II) as well as their correlation with angiogenesis and prognosis in gastric carcinoma.

METHODSHIF-1alpha and IGF-II mRNA expression were analyzed using in situ hybridization, microvessel density (MVD) was determined by anti-CD34 immunostaining in 118 cases gastric carcinoma.

RESULTSThe positive rates of HIF-1alpha mRNA and IGF-II mRNA were 49.2% and 47.4%, respectively. In stage T3-T4 cases, positive rates of HIF-1alpha and IGF-II mRNA expression, the frequencies of vessel invasion, lymph node and distant metastasis were significantly higher than those in stage T1-T2 cases. The mean MVD in stage T3-T4 tumors, vessel invasion, lymph node metastasis and distant metastasis were significantly more frequent than those in stage T1-T2 tumors. The mean MVD in tumors with positive HIF-1alpha and IGF-II mRNA expression was significantly higher than those in tumors without HIF-1alpha and IGF-II mRNA expression. The expression of HIF-1alpha was positively correlated with IGF-II mRNA. There were positive correlations between MVD and expression of HIF-1alpha and IGF-II mRNA, respectively. The mean survival time and 5-year survival rate in cases with positive HIF-1alpha and VEGF expression and MVD value >/= 41.5 were significantly shorter than those in cases with negative HIF-1alpha and VEGF expression and MVD value < 41.5.

CONCLUSIONSHIF-1alpha and IGF-II play an important role in tumor invasion and metastasis, especially in tumor angiogenesis. So test of the expression of HIF-1alpha and IGF-II may act as a useful index of treatment and prognosis.

Adult ; Aged ; Antigens, CD34 ; analysis ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Immunohistochemistry ; In Situ Hybridization ; Insulin-Like Growth Factor II ; metabolism ; Male ; Middle Aged ; Neovascularization, Pathologic ; genetics ; metabolism ; pathology ; Prognosis ; RNA, Messenger ; genetics ; metabolism ; Stomach Neoplasms ; blood supply ; genetics ; metabolism ; Survival Analysis

OBJECTIVETo investigate mRNA expression of syndecan-1 and heparanase in gastric carcinoma, and their correlation with the growth-pattern, invasion, metastasis and prognosis of gastric carcinoma.

METHODSIn situ hybridization technique was used to examine mRNA expression of syndecan-1 and heparanase in 118 specimens of gastric carcinoma.

RESULTSThe positive rates of syndecan-1 mRNA and heparanase mRNA were 42.4% and 55.9%, respectively. The expression of syndecan-1 mRNA and heparanase mRNA were related to tumor invasion depth (chi(2) = 32.95, P = 0.001; chi(2) = 23.19, P = 0.001), vessel invasion (chi(2) = 46.22, P = 0.001; chi(2) = 33.78, P = 0.001), lymph node (chi(2) = 28.62, P = 0.001; chi(2) = 25.43, P = 0.001) and distant metastasis (chi(2) = 63.30, P = 0.001; chi(2) = 65.76, P = 0.001), and syndecan-1 mRNA positive expression was related to tumor size (chi(2) = 6.25, P = 0.012). There was a negative relationship between Syndecan-1 mRNA and heparanase mRNA expression (r = -0.844, P = 0.001). The mean survival time of cases with low expression of syndecan-1 mRNA was significantly shorter than that of cases with high expression (r = 36.48, P = 0.001), and meanwhile, the mean survival time of heparanase mRNA positive cases was significantly shorter than that of cases with negative expression (r = 34.41, P = 0.001).

CONCLUSIONSThe mRNA expression of syndecan-1 and heparanase can predict the invasion and metastasis of gastric carcinoma, and can be used as markers of prognosis of gastric carcinoma.

Adult ; Aged ; Female ; Follow-Up Studies ; Glucuronidase ; genetics ; metabolism ; Humans ; In Situ Hybridization ; Lymphatic Metastasis ; Male ; Middle Aged ; RNA, Messenger ; genetics ; Stomach Neoplasms ; metabolism ; mortality ; pathology ; Survival Rate ; Syndecans ; genetics ; metabolism

BACKGROUNDStudies have reported that patients with human immunodeficiency virus (HIV) have a high incidence of osteonecrosis of the femoral head (ONFH). Total hip arthroplasty (THA) is an effective management of ONFH. However, little data exist regarding the use of THA for the HIV patients with ONFH in China. This study reviewed the outcomes of HIV-positive patients who underwent THA for ONFH, compared with HIV-negative individuals.

METHODSThe patients who underwent THA for ONFH from September 2012 to September 2014 in Beijing Ditan Hospital, Capital Medical University were retrospectively studied. Twenty-eight HIV-positive patients and 35 HIV-negative patients underwent 48 THAs and 45 THAs with cementless components, respectively. Medical records and follow-up data were reviewed. Harris Hip Score (HHS) was applied to evaluate the pain and function of the hips before and after THA. Complications such as wound healing, surgical site infection, deep venous thrombosis, pulmonary embolism, sepsis, mortality, and complications from the prosthesis were reviewed. The operation time, blood loss, and hospital stay were compared between the two groups.

RESULTSThe mean follow-up period was 19.5 ± 5.8 months (ranging from 6 to 30 months). The mean age of the HIV-positive patients with osteonecrosis at the time of surgery was 35 years old, which was significantly lower than that of the HIV-negative group (42 years old) (P < 0.05). The HIV-positive patients underwent surgery a mean of 2.5 years after their original symptoms, which was significantly shorter than the HIV-negatives' (mean 4 years) (P < 0.05). Among HIV-positive patients, the prevalence of being male and rate of bilateral procedures were significantly higher than those in the HIV-negative group (P < 0.05). The operation time in HIV-positive patients was significantly longer than that in HIV-negative patients (P < 0.05). There were no significant differences in blood loss or hospital stay between the two groups (P > 0.05). The HHSs of two groups significantly improved after THAs (P < 0.05), without significant difference between two groups. No wound complication, sepsis, mortality, prosthesis complication, and occupational exposure occurred, except for two cases of heterotopic ossification and one case of humeral head necrosis.

CONCLUSIONSONFH is more likely to occur bilaterally in younger HIV-positive males. The development of osteonecrosis seems faster in HIV-positive patients than in HIV-negative patients. This should be cautionary for asymptomatic HIV-positive patients with low viral RNA level and in the primary HIV stage. Despite longer operation times in the HIV-positive patients than in the HIV-negative patients, THA is still a safe and efficient approach to treat ONFH in HIV-positive patients. The incidence of complications is much lower than previously reported. However, the long-term follow-up is needed.

Adult ; Arthroplasty, Replacement, Hip ; methods ; China ; Female ; Femur Head Necrosis ; pathology ; surgery ; HIV Infections ; surgery ; Humans ; Magnetic Resonance Imaging ; Male ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo investigate effects of developmental lead exposure on nitric oxide synthase (NOS) activity in different brain regions and on N-methyl-D-aspartate (NMDA) receptor mRNA expression in the hippocampus of rats. On the basis of these observations, we explored possible mechanisms by which lead exposure leads to impaired learning and memorizing abilities in children.

METHODSA series of rat animal models exposed to low levels of lead during the developing period was established (drinking water containing 0.025%, 0.05% and 0.075% lead acetate). NOS activities in the hippocampus, the cerebral cortex, the cerebellum and the brain stem were determined with fluorescence measurement and levels of mRNA expression of the NMDA receptor 2A (NR2A) subunit and NMDA receptor 2B (NR2B) subunit in the rat hippocampus were measured with Retro-translation (RT-PCR).

RESULTSThere were no differences in the body weight of rat pups between any of the groups at any given time (P>0.05). The blood lead level of Pb-exposed rat pups showed a systematic pattern of change: at 14 d of age, it was lower than that at 7 d of age, then rising to the peak level at 21 d and finally falling to lower levels at 28 d. The hippocampal NOS activities of lead-exposed groups were all lower than that of the control group on the 21st and 28th day (P<0.01). NOS activities in the cerebellum of lead-exposed groups were all lower than that of the control group on the 21st and 28th day (P<0.001) and the NOS activity of the 0.025% group was significantly lower than that of the 0.05% and 0.075% groups on the 28th day (P<0.05). NOS activity in the cerebral cortex of the 0.075% group was significantly lower than that of the control, 0.025% and 0.05% groups on the four day spans (P<0.001). There was no significant difference of NOS activity in the brain stem between any lead-exposed group and the control group on the four day spans. In the 0.05% and the 0.075% groups, the level of NR2A mRNA expression was higher than that in the control group at 7 d and 14 d of age (P<0.05). In the 0.025% group, the level of NR2A was found to be higher than that in the control group at 7 d of age only (P<0.05). No significant differences were found for the levels of NR2B mRNA expression between any of the groups at any given time.

CONCLUSIONSNOS activity in the hippocampus, the cerebral cortex and the cerebellum are inhibited by lead exposure. The degree of the inhibitory effect depends on the time span of exposure and the lead concentration. Developmental low-level lead exposure was found to raise the level of NR2A mRNA expression in the hippocampus of rats. Developmental low-level lead exposure does not affect the level of NR2B mRNA expression in the hippocampus.

Animals ; Animals, Newborn ; Brain ; drug effects ; growth & development ; metabolism ; Environmental Exposure ; adverse effects ; Enzyme Activation ; drug effects ; Female ; Lead ; toxicity ; Male ; Neurotoxins ; toxicity ; Nitric Oxide Synthase ; metabolism ; Rats ; Receptors, N-Methyl-D-Aspartate ; metabolism

A full-length cDNA of GGPPS gene from Tripterygium wilfordii suspension cells was obtained by use of RACE strategy (GeneBank: KM978333), and then analyzed by bioinformatics approaches. TwGGPPS cDNA has 1857 nucleotides and an open reading frame (ORF) encoding a protein of 514 amino acid residues. The deduced protein has isoelectric point (pI) of 7.85, a calculated molecular weight about 57.13 kD, 5 conserved domains and 2 functional domains. PSORT Prediction showed it was located at plasma membrane. Phylogenetic analysis demonstrated that TwGGPPS1 was similar to GGPPS from other species of plants. For the first time the cloning of geranylgeranyl diphosphate synthase gene from T. wilfordii was reported, it lays the foundation for further research of diterpenoids biosynthetic pathway.
Amino Acid Sequence ; Cloning, Molecular ; Farnesyltranstransferase ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Sequence Alignment ; Sequence Homology, Amino Acid ; Tripterygium ; chemistry ; enzymology ; genetics

4-(Cytidine-5-diphospho) -2-C-methyl-D-erythritol kinase is a key enzyme in the biosynthesis pathway of terpenoids. According to the transcriptome database, the specific primers were designed and used in PCR. The bioinformatic analysis of the sequenced TwCMK gene was performed in several bioinformatics software. The Real-time fluorescence quantification polymerase chain reaction (RT-qPCR) were used to detect the expression levels of TwCMK from T. wilfordii after elicitor MeJA supplied. The results showed that the full length of TwCMK cDNA was 1 732 bp encoding 387 amino acids. The theoretical isoelectric point of the putative TwCMK protein was 5.79 and the molecular weight was about 42.85 kDa. MeJA stimulated the rising of TwCMK expression in suspension cell and signally impacted at 24 h. The research provides a basis for further study on the regulation of terpenoid secondary metabolism and biological synthesis.
Amino Acid Sequence ; Cloning, Molecular ; Computational Biology ; Gene Expression Regulation, Plant ; Models, Molecular ; Molecular Sequence Data ; Phosphotransferases (Alcohol Group Acceptor) ; chemistry ; genetics ; metabolism ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Sequence Alignment ; Tripterygium ; chemistry ; enzymology ; genetics

OBJECTIVETo investigate the effects of atorvastatin on parathyroid hormone 1-34 (PTH1-34) induced neonatal rat cardiomyocytes hypertrophy and on the expression changes of small GTP-binding protein (K-Ras) and extracellular signal regulated protein kinases 1/2 (ERK1/2).

METHODSNeonatal rat cardiomyocytes hypertrophy was established with 10(-7) mol/L rPTH1-34 in the presence or absence of 10(-5) mol/L atorvastatin or 10(-4) mol/L mevalonic acid (MVA). Cardiomyocyte diameter was measured by Motic Images Advanced 3.0 software, the synthetic rate of protein in cardiomyocytes was determined by (3)H-leucine incorporation and single-cell protein content was measured by BCA. The concentration of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) were determined by ELISA. Protein expression of ERK1/2, p-ERK1/2 and K-Ras was detected by Western blot.

RESULTSCompared to PTH1-34 group, cellular diameter was decreased 12.07 µm, (3)H-leucine incorporation decreased 1622 cpm/well and single-cell protein content decreased 84.34 pg, ANP or BNP concentration reduced 7.13 µg/L or 20.04 µg/L, protein expression of K-Ras, ERK1/2 or p-ERK1/2 downregulated 0.81, 0.19 and 1.44 fold, respectively, in PTH1-34 plus atrovastatin co-treated cardiomyocytes (all P < 0.05). Compared to PTH1-34 plus atrovastatin co-treated group, cardiomyocyte diameter increased 4.95 µm, (3)H-leucine incorporation increased 750 cpm/well and single-cell protein content increased 49.08 pg, ANP or BNP increased 3.12 µg/L or 9.35 µg/L and protein expression of K-Ras, ERK1/2 or p-ERK1/2 upregulated 0.52, 0.06 and 1.19 fold (all P < 0.05) in MVA, PTH1-34 and atrovastatin co-treated cardiomyocytes.

CONCLUSIONSAtrovastatin attenuates PTH1-34 induced neonatal rat cardiomyocytes hypertrophy through downregulating K-Ras and ERK1/2 pathway.

Animals ; Animals, Newborn ; Atorvastatin Calcium ; Cardiomegaly ; drug therapy ; metabolism ; Cells, Cultured ; Heptanoic Acids ; pharmacology ; therapeutic use ; MAP Kinase Signaling System ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Myocytes, Cardiac ; drug effects ; metabolism ; Parathyroid Hormone ; metabolism ; Pyrroles ; pharmacology ; therapeutic use ; Rats ; Rats, Wistar

AIM: To observe thickness and morphological changes of bulbar conjunctiva pre- and post epidemic hemorrhagic conjunctivitis ( EHC ) therapy by optical coherence tomography ( OCT) .
METHODS: Observed morphological changes and measured the bulbar conjunctiva thicknesses of 29 cases (36 eyes) of incipient (1-2d) EHC patients, who were received and treated by department of ophthalmology, the Putuo Affiliated Hospital of Shanghai University of Traditional Chinese Medicine from May 2013 to December 2013, by OCT. Then measured the thickness again on 7, 14d after the therapy.
RESULTS: Among 29 patients (36 eyes), 7d after the EHC therapy, in 27 cured eyes, the full-thickness ( before 344. 00±59. 91μm, after 230. 19±22. 16μm, t=11. 75, P<0-01); epithelial thickness ( before 56. 52±6. 19μm, after 51. 37±5. 53μm, t=4. 61, P<0. 01); and stromal thickness (before 287. 11±60. 56μm, after 178. 81±20. 20μm, t=10. 69, P<0. 01) of patients' bulbar conjunctiva were thicker than values measured after therapy with significant difference. Significant difference was also found for full-thickness ( before 361. 39±65. 56μm, after 233. 44±22. 57μm, the difference was statistically significant, t=14. 45, P<0. 01);epithelial thickness ( before 55. 50±6. 72μm, after 46. 67±5-24μm, t=10. 06, P<0. 01) and stromal thickness ( before 305. 61±66. 02μm, after 186. 78±21. 82μm, t=13. 11, P<0-01 ) of patients' bulbar conjunctiva between values measured before and 14d after therapy.
CONCLUSION: The OCT is able to measure the thickness of bulbar conjunctiva in EHC patients. An significant increase was found in full, epithelial and stromal thickness of EHC patients' bulbar conjunctiva. With recovery from the disease, subepithelial fluid, interlaminar fluid and edema of the bulbar conjunctival stroma faded away firstly, which provide references for clinical therapies of the EHC.