Objective: To investigate the effect of MeJA combined with high temperature stress in the treatment for the accumulation of triterpenoids in the birch (Betula platyphylla) suspension cells. Methods: After MeJA (25, 50, 100, and 150 μmol/L)and high temperature (50℃ for 2 h) treatment, the cell growth, viability, content of MDA, the activity of defense enzyme, total triterpenoids content, and the gene expression levels of triterpenoids synthesis were measured. Results: The combination of high temperature stress and MeJA treatment had a more powerful positive effect on the synthesis of triterpenoids than single MeJA or high temperature treatment in birch cells. Moreover, the concentration of total triterpenoids had the highest level when adding 150 μmol/L MeJA after the high temperature processing, was up to 76.6 mg/g, which was 81.3%, 159.9% and 13.1% higher than those in the blank control, individual MeJA treatment or the heat treatment alone respectively. Meanwhile, the gene expression levels of SS, SE, BPW, and BPY, related to the triterpenoids synthesis, had an increase about 297.1%, 83.7%, 1 032.6%, and 282.4% compare to the control. The MeJA after high temperature treatment enhanced the activity of SOD and PAL compared with the control, inhibited the cell growth and viability. Conclusion: The treatment of MeJA after high temperature affects the cell growth, viability, and activity of defense enzyme, regulates the genes expression level of triterpenoids synthesis, and eventually could make cells to produce the triterpenoids substance effectively.

Afferent Pathways ; drug effects ; Animals ; Freund's Adjuvant ; adverse effects ; Ganglia, Sensory ; drug effects ; Hyperalgesia ; chemically induced ; Inflammation ; Male ; Morphine ; pharmacology ; Rats ; Rats, Sprague-Dawley

The zinc chloride and cystine resistant strain of S.cerevisiae YZM-14(ZnCl2r,Cysr) was screened with the mutant processing of the protoplast of S.cerevisiae by combinative mutagens of ultraviolet and nitrite.The glutathione(GSH) production(84.72 mg/L),dry cell weight(7.63 g/L) and the intracellular GSH content(11.10 mg/g) of YZM-14 were 2.79,1.63 and 1.71 times compared with that of the initial strain.The biosynthetic process of GSH was divided into three phases according to the time course of the specific cell growth rate and GSH yielding coefficient.In the second phase,the metabolic flux of the pentose phosphate pathway and the GSH precursors biosynthetic pathway of the mutant strain increased by 8.1 mmol/(g?h),compared with that of the initial strain.Furthermore,the metabolic flux of the organic acids secretion of the mutant strain decreased.Through these mechanisms,the utilization efficiency of the carbon sources was enhanced and high production of GSH was obtained.

Objective To study the influence and dose effect of ultrasound on the contraction of uterine smooth muscle in rats.Methods Estradiol benzoate was injected into rats three days before conducting an in-vitro experiment.Their uteri were resected and irradiated with ultrasound(0.8 MHz,3 W/cm~2,0-40 rain).The contrac- tion frequency and amplitude were recorded using an MS-302 biological experiment system.Results It could be seen that the contraction frequency and amplitude,and general contractile activity were significantly increased during ultrasonic irradiation(P＜0.01).The increased contraction frequency and amplitude lasted for ten minutes,and then the normal contraction pattern resumed.The contraction frequency as well as the percentage change in eontraction fre- quency were highest during the first 15 minutes of ultrasonic irradiation;the contraction amplitude as well as the per- centage change in amplitude were highest during 40 minutes of ultrasonic irradiation.Contraction activity was at its highest for 30 minutes,but the percentage change in activity was highest for 20 minutes.Conclusions Ultrasound can induce uterine smooth muscle contraction in rats.This biological effect is related to the irradiation time.

BACKGROUNDCancer of the esophagus and gastroesophageal junction remains a virulent malignancy with poor prognosis. Rapid progresses were made in chemotherapeutic agents and the development of molecular markers allowed better identification of candidates for targeted therapy. This study aimed to identify the candidate peptides used for anti-angiogenic therapy of esophageal cancer by in vivo screening C7C peptide library for peptides binding specifically to blood vessels of human esophageal cancer.

METHODSThe phage displayed C7C peptide library was injected intravenously into mice bearing human esophageal tumor xenografts under renal capsule. After 5 rounds of screening, 13 clones were picked up individually and sequenced. During each round of screening, titers of phage recovery were calculated from tumor xenograft and control tissues. Homing of these 9 peptides to tumor vessel was detected by calculating phage titers in the tumor xenograft and control tissues (lung and spleen) after each phage was injected into mice model, and compared with the distribution of phage M13 and VIII-related antigen in tumor xenograft by immunohistochemical staining. Comparisons among groups of data were made using one-way analysis of variance (ANOVA), followed by the Bonferroni multiple comparisons test.

RESULTSThe number of phage recovered from tumor tissue of each round increased gradually in tumor group while decreased in control groups (P < 0.01 in tumor and spleen, P < 0.05 in lung). Immunohistochemical staining showed similar staining pattern with M13 antibody or VIII-related antigen antibody, suggesting that phages displaying the selected peptides could home to blood vessel of human esophageal cancer. According to their DNA, 9 corresponding peptide sequences were deduced. And the homing ability to blood vessel of phages displaying the selected peptides was confirmed by comparing with their recovery in tumor and control tissues. Two motifs, YSXNXW and PXNXXN, were also obtained by analyzing the homology of these peptide sequences. The staining distribution of phage with the sequence of PNPNNST was similar to that of the blood vessel marker factor VIII-related antigen staining. After sequencing, each phage with the selected peptide of PNPNNST with 1.0 × 10(11) pfu/ml was injected intravenously into mice. The homing ability to tumor vessel of these 9 kinds of peptides in the xenograft was higher than control tissues (lung and spleen).

CONCLUSIONNine peptides obtained from in vivo screening homed to the blood vessel of human esophageal cancer, and the two motifs of YSXNXW and PXNXXN are the possible biochemical recognition units binding to vascular endothelial cells of esophageal cancer.

Animals ; Antineoplastic Agents ; therapeutic use ; Endothelial Cells ; drug effects ; Esophageal Neoplasms ; blood supply ; drug therapy ; metabolism ; Humans ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Peptide Library ; Peptides ; therapeutic use

Objective To observe the effect of Tetramethylpyrazine combined with cisplatin on Lewis lung adenocarcinoma by the change of hypoxia-inducible factor 1 alpha ( HIF -1α) , basic fibroblast growth factorb ( b-FGF) ,vasohibin-1 ( VASH-1 ) in mice tumor tissues and explore the mechanism of anti -tumor.Methods Forty -eight mice were randomly divided into 4 groups, control group, cisplatin group, Tetramethylpyrazine group and combined group, 12 rats in each group, cisplatin at a dose of 2 mg· kg -1 , Tetramethylpyrazine at a dose of 100 mg· kg-1 , combination group( the doses as above).All mice were sac-rificed after treatment for two weeks and these tumors were obtained.The weighed, and the tumor inhibitory was calculated.The expressions of HIF-1α, b -FGF and VASH -1 were determined by immunohisto-chemistry after two weeks administration.Results Compared with the control group, the tumor inhibition rate in experimental groups was signi-ficantly increased ( P<0.05 ) , and was highest in combination group.The expression of HIF -1α, b -FGF and VASH -1 in the cisplatin group, Tetramethylpyrazine group and combined group were decreased obviously compared with control group( P<0.05).And the levels of the expression were the lowest in the combined group and the highest in control group.HIF -1αis positively correlated with b -FGF ( P <0.05 ) , b -FGF is positively correlated with VASH-1 ( P <0.05 ) , HIF-1αis uncorrelated with VASH-1.Conclusion Tetramethylpyrazine combined with cisplatin decreased the expression of HIF-1α, thereby reducing the expression of b-FGF and VASH-1 , inhibiting the tumor growth synergistically.

OBJECTIVESTo study the correlation between Chlamydiae pneumonia and carotid atherosclerosis, and the correlation between the infection of Chlamydiae pneumonia and ischemic events.

METHODSThe study group consisted of 19 patients who underwent unilateral carotid endarterectomy surgery during the period from January 2010 to December 2011, and the atherosclerotic plaque specimens were harvested from these patients. The control group consisted of 10 patients who underwent extracranial-intracranial bypass surgery during the same period, and the normal external carotid artery specimens were got from these patients. The clinical data between the two groups had comparability. The presence of Chlamydiae pneumonia in atherosclerotic plaque and normal artery tissue were investigated by immunohistochemistry. The expression of Toll-like receptor 2 (TLR2), tumor necrosis factor-α (TNF-α) and vascular cell adhesion molecule-1 (VCAM-1) in the atherosclerotic plaque infected with Chlamydiae pneumonia were also detected. Data were analyzed using Fisher's exact test.

RESULTChlamydiae pneumonia was found in 9 of 19 atherosclerotic plaques, while no positive result was found in control group. The statistical analysis showed a significant difference (P = 0.011). Among the 19 patients in study group, 15 of them had ischemic events, and Chlamydiae pneumonia was found in 9 of these 15 patients; while the other 4 patients didn't have any ischemic events and no Chlamydiae pneumonia was found in them, but there was no statistical different between them (P = 0.087). Through immunohistochemistry, the expression of Chlamydiae pneumonia, TLR2, TNF-α and VCAM-1 were found in same area.

CONCLUSIONSThere is a correlation between Chlamydiae pneumonia and carotid atherosclerosis.And there might be a correlation between Chlamydiae pneumonia and cerebral ischemic events.

Carotid Artery Diseases ; metabolism ; microbiology ; Case-Control Studies ; Chlamydophila Infections ; complications ; Chlamydophila pneumoniae ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Plaque, Atherosclerotic ; microbiology ; Toll-Like Receptor 2 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo explore the features of proton magnetic resonance spectroscopy (1H-MRS) of the hippocampus in schizophrenia patients before and after stereotactic neurosurgery.

METHODS1H-MRS was performed to determine NAA/Cr and CHO/Cr ratios on the bilateral hippocampal regions before and after stereotactic neurosurgery in 20 schizophrenia patients, with 20 healthy individuals as the controls.

RESULTSThe NAA/Cr ratio in the hippocampal regions was significantly lower and the CHO/Cr ratio significantly higher in schizophrenia patients before the surgery than in the healthy controls (P<0.01). The NAA/Cr and CHO/Cr ratios in the hippocampal regions underwent no significant changes in the patients after the surgeries (P>0.05).

CONCLUSIONNeuronal and cell membrane metabolism impairment is present in the hippocampus of schizophrenia patients, and stereotactic neurosurgery does not produce obvious adverse effects on the cell membrane metabolism in the hippocampus of the patients.

Adolescent ; Adult ; Aspartic Acid ; analogs & derivatives ; metabolism ; Case-Control Studies ; Choline ; metabolism ; Creatine ; metabolism ; Female ; Hippocampus ; metabolism ; pathology ; Humans ; Magnetic Resonance Spectroscopy ; methods ; Male ; Protons ; Schizophrenia ; metabolism ; pathology ; surgery ; Stereotaxic Techniques ; Young Adult

Molecular biological studies and electrophysiological data have demonstrated that acetylcholine (ACh) is the principal cochlear and vestibular efferent neurotransmitter among mammalians. However, the functional roles of ACh in type II vestibular hair cells among mammalians are still unclear, with the exception of the well-known alpha9-containing nicotinic ACh receptor (alpha9-nAChR) in cochlear hair cells and frog saccular hair cells. In this study, the properties of the ACh-sensitive current were investigated by whole-cell patch clamp technique in isolated type II vestibular hair cells of guinea pigs. The direct effect of extracellular ACh was to induce a hyperpolarization effect in type II vestibular hair cells. Type II vestibular hair cells displayed a sustained outward current in response to the perfusion of ACh. It took about 60 s for the ACh-sensitive current to get a complete re-activation. The reversal potential of the ACh-sensitive current was (-66 +/- 8) mV, which indicated that potassium ion was the main carrier of this current. The blocking effect by the submillimolar concentration of tetraethylammonium (TEA) further indicated that extracellular ACh stimulated the calcium-dependent potassium current. Following replacement of the compartment of NaCl in the normal external solution with TrisCl, LiCl or saccharose respectively, the amplitude of the ACh-sensitive current was not affected. Blocking of the release of intracellular Ca(2+) stores by intracellular application of heparin failed to inhibit the ACh-sensitive current. Therefore, extracellular Na(+)and the inositol 1,4,5-trisphosphate (IP(3))-dependent intracellular Ca(2+)release were not involved in the activation of the ACh-sensitive current. However, the ACh-sensitive current was strongly affected by the concentration of the extracellular K(+), extracellular Ca(2+) and intracellular Mg(2+). The amplitude of the ACh- sensitive current was strongly inhibited by high concentration of extracellular K(+). In the Ca(2+)-free external solution, ACh only activated a very small current; however, the ACh-sensitive current demonstrated a Ca(2+)-dependent inhibition effect in high concentration of Ca(2+)solution. In addition, the ACh-sensitive current was inhibited by increasing of the concentration of intracellular Mg(2+). In conclusion, the present results demonstrate that ACh plays an important role in the vestibular efferent system. The fact that Na(+) is not involved in the ACh-sensitive current will not favor the well-known profile of alpha9-nAChR, which is reported to display a small but important permeability to Na(+). It is also suggested that, in vivo, the amplitude of the ACh-induced hyperpolarization may strongly depend on the concentration of extracellular Ca(2+)and intracellular Mg(2+).
Acetylcholine ; physiology ; Animals ; Calcium ; physiology ; Guinea Pigs ; Hair Cells, Vestibular ; physiology ; Magnesium ; physiology ; Patch-Clamp Techniques ; Potassium Channels, Calcium-Activated ; physiology

BACKGROUNDTuberculosis remains the leading cause of human death. Currently, Bacillus Calmette-Guérin (BCG) is the only available vaccine against tuberculosis but its efficacy is highly variable. Thus, developing new tuberculosis vaccines becomes an urgent task. In this study, we evaluated in BALB/c mice the humoral and cellular immune responses of recombinant BCG expressing the antigen ESAT-6 from Mycobacterium tuberculosis.

METHODSEscherichia coli-BCG shuttle plasmid named pDE22-esat-6 was constructed by inserting the BamHI/EcoRI digested esat-6 gene PCR product into the similarly digested parental plasmid pDE22. BCG cells were transformed with pDE22-esat-6, which was named recombinant BCG (rBCG). BALB/c mice were immunized subcutaneously on the back with 100 microl normal saline containing 10(6) CFU of BCG or rBCG. They were sacrificed after 4 weeks to detect their humoral and cellular responses.

RESULTSThere was no any significant differences in the growth characteristics between the conventional BCG and rBCG. In immunized mice, the IgG antibody titres of rBCG group were as high as 1:8000, which was significantly higher than that in BCG group (1:1400, P < 0.05). The elicited IFN-gamma level of rBCG group was (1993 +/- 106) pg/ml, which was also significantly higher than that in BCG group ((1463 +/- 105) pg/ml, P < 0.05). The splenocyte proliferation index of rBCG group reached 4.34 +/- 0.31, which was higher than that of BCG group (3.79 +/- 0.24, P < 0.05).

CONCLUSIONrBCG secreted expressing antigen ESAT-6 stimulated stronger humoral and cellular immune responses than BCG did, and, therefore may be the better vaccine against mycobacterium tuberculosis.

Animals ; Antigens, Bacterial ; genetics ; immunology ; BCG Vaccine ; immunology ; Bacterial Proteins ; genetics ; immunology ; Interferon-gamma ; biosynthesis ; Lymphocyte Activation ; Male ; Mice ; Mice, Inbred BALB C ; Mycobacterium tuberculosis ; immunology ; Recombinant Proteins ; immunology ; Vaccines, Synthetic ; immunology