Chemical constituents of flowers and fruits of Rabdosia excisa / 中国天然药物

AIM@#To study the chemical constituents in the flowers and fruits of Rabdosia excisa.@*METHODS@#The compounds were isolated and purified by silica gel column chromatography and their structures were identified by spectroscopic methods.@*RESULTS@#Sixteen compounds isolated from the flowers and fruits of this plant were identified as: stigm asterol (I), α-amyrin palmitate (II), ursolic acid (III), 2α, 3α, 19-trihydroxy-urs-12-en-28-oic acid (IV), 2α-hydroxyursolic acid (V), maslinic acid (VI), isodonal (VII), maoyecrystal E (VIII), kamebakaurin (XI), macrocalyxin G (X), epinodosinol (XI), rabdosichuanin C (XII), kamebacetal A (XIII), oridonin (XIV), enmenol-glucoside (XV), and lasiononin (XVI).@*CONCLUSION@#All the constituents were found in Rabdosia excisa for the first time, except constituents III, IX, XII and XIV.

Long-term stable expression of antisense cDNA of cyclin B1 profoundly inhibits the proliferation of tumor cells and suppresses tumorigenicity in implanted mice / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Cyclin B1 (CLB1) is necessary for mitotic initiation in mammalian cells and plays important roles in cancer development. Therefore, a potential strategy in cancer therapy is to suppress the activity of CLB1 by delivering antisense constructs of CLB1 into tumor cells. In previous CLB1 studies, antisense constructs with a short half life were often used and these constructs might not persistently inhibit CLB1.</p><p><b>METHODS</b>We successfully created a recombinant plasmid encoding the full-length antisense cDNA of mouse cyclin B1 (AS-mCLB1) and transfected this construct to the murine Lewis lung carcinoma (LL/2) and CT-26 colon carcinoma (CT-26) cells. We isolated clones of LL/2 and CT-26 transfectants with stable expression of AS-mCLB1. Reverse transcriptional polymerase chain reaction (RT-PCR) and Western blot were applied to detect the expression of the mRNA and protein levels of CLB1. To further test the efficacy of this strategy in vivo, AS-mCLB1-expressing LL/2 and CT-26 transfectants were implanted into mice.</p><p><b>RESULTS</b>We found the expression of the mRNA and protein levels of CLB1 decrease in these transfectants. The inhibition of CLB1 caused prominent G1 arrest, abnormal morphology, retarded cell growth and an increase in apoptosis. In AS-mCLB1-expressing LL/2 and CT-26 transfectants implanted mice, tumorigenicity was effectively suppressed compared with the controls. In addition, the expression of AS-mCLB1 also significantly increases the survival duration of implanted animals.</p><p><b>CONCLUSION</b>AS-mCLB1 is likely to be useful in future cancer therapy, which may be associated with its ability to down-regulate the expression of CLB1 and then induce G1arrest and apoptosis in tumor cells.</p>

Effect of intestinal lymphatic pathway on free radical and inflammatory mediator of myocardium in shock rats / 中国应用生理学杂志

<p><b>AIM</b>To observe the effect of mesenteric lymph duct ligation on free radical and inflammatory mediator of myocardium with severe hemorrhagic shock in rats at different period, and explore the effect of intestinal lymphatic pathway on myocardium injury pathogenesis in shock rats.</p><p><b>METHODS</b>78 male Wistar rats were divided into the sham group, shock group and ligation group. The model of serious hemorrhagic shock was established in shock group, ligation group, and mesenteric lymph was blocked by ligating mesenteric lymph duct in ligation group after resuscitate. All rats were executed and taken out heart making for homogenate of 10 percent to determine the MDA, SOD, tumor necrosis factor-alpha (TNFalpha), interleukin-6 (IL-6), myeloperoxidase (MPO), NO and NOS at after shock 90 min, after transfusion and resuscitate 0 h, 1 h, 3 h, 6 h, 12 h and 24 h etc. different times, and the expression of inducible nitric oxide synthase (iNOS) mRNA in myocardium was detected by RT-PCR.</p><p><b>RESULTS</b>The contents of MDA, TNFalpha, IL-6, MPO, NO, NOS and iNOS expression in myocardium of shock group were rising after transfusion and resuscitate, and that was higher level at 3 h to 12 h, and that was significantly higher than sham group, the activity of SOD was significantly lower than sham group. The contents of MDA, TNFalpha, IL-6, MPO, NO, NOS and iNOS expression in myocardium of ligation group were significantly lower than that of shock group at sameness points, and the SOD activity was higher.</p><p><b>CONCLUSION</b>The mesenteric lymph duct ligation and blocking mesenteric lymph could reduce the PMN detaining, decrease the discharging of TNFa and IL-6, reduce the NO and expression of iNOS mRNA, and reduce the releasing of free radical and consuming of SOD.</p>

Edaravone protects PC12 cells from ischemic-like injury via attenuating the damage to mitochondria / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>BACKGROUND</b>Edaravone had been validated to effectively protect against ischemic injuries. In this study, we investigated the protective effect of edaravone by observing the effects on anti-apoptosis, regulation of Bcl-2/Bax protein expression and recovering from damage to mitochondria after OGD (oxygen-glucose deprivation)-reperfusion.</p><p><b>METHODS</b>Viability of PC12 cells which were injured at different time of OGD injury, was quantified by measuring MTT (2-(4,5-dimethylthia-zol-2-yl)-2,5-diphenyltetrazolium bromide) staining. In addition, PC12 cells' viability was also quantified after their preincubation in different concentration of edaravone for 30 min followed by (OGD). Furthermore, apoptotic population of PC12 cells that reinsulted from OGD-reperfusion with or without preincubation with edaravone was determined by flow cytometer analysis, electron microscope and Hoechst/PI staining. Finally, change of Bcl-2/Bax protein expression was detected by Western blot.</p><p><b>RESULTS</b>(1) The viability of PC12 cells decreased with time (1 - 12 h) after OGD. We regarded the model of OGD 2 h, then replacing DMEM (Dulbecco's Modified Eagle's Medium) for another 24 h as an OGD-reperfusion in this research. Furthermore, most PC12 cells were in the state of apoptosis after OGD-reperfusion. (2) The viability of PC12 cells preincubated with edaravone at high concentrations (1, 0.1, 0.01 micromol/L) increased significantly with edaravone protecting PC12 cells from apoptosis after OGD-reperfusion injury. (3) Furthermore, edaravone attenuates the damage of OGD-reperfusion on mitochondria and regulated Bcl-2/Bax protein imbalance expression after OGD-reperfusion.</p><p><b>CONCLUSION</b>Neuroprotective effects of edaravone on ischemic or other brain injuries may be partly mediated through inhibition of Bcl-2/Bax apoptotic pathways by recovering from the damage of mitochondria.</p>

Effects of nitric oxide on peritoneal lymphatic stomata and lymph drainage via NO-cGMP-Ca2+ pathway / 生理学报

To study the cell signal transduction mechanism of nitric oxide (NO) on the peritoneal lymphatic stomata and lymph drainage in the rat, cGMP content were measured by a commercially available radioimmunoassay kit, and the [Ca(2+)](i) were observed by a confocal laser scanning microscope in the cultured peritoneal mesothelial cell. Animal experiment was practiced to study the effect of NO-cGMP-Ca(2+) pathway on the lymphatic stomata and lymph absorption. The results showed that: (1) Sper/NO increased cGMP of the rat peritoneal mesothelial cell (RPMC) in a dose-dependent manner (P<0.01) compared to the control group. This effect was blocked by 1H-[1,2,4] oxadiazolo [4,3-a] quinoxalin-1-one (ODQ) (P<0.05), a specific inhibitor of soluble guanylyl cyclase (sGC). The level of [Ca(2+)](i) in single RPMC decreased by adding Sper/NO (P<0.05). Pretreatment with ODQ for 10 min blocked the Sper/NO-induced decrease in [Ca(2+)](i). L-typed calcium channel blocker nifedipine induced an immediate and marked decrease in [Ca(2+)](i) (P<0.05).. After [Ca(2+)](i) reached a balance again, adding Sper/NO could not change [Ca(2+)](i) (P>0.05). (2) Sper/NO increased the area of the stomata (P<0.01) and the quantity of the tracer in a dose-dependent manner (P<0.05) compared to the control group. Pretreatment with ODQ significantly inhibited Sper/NO-induced change of lymphatic stomata and lymph drainage (P<0.01). Nifedipine increased the opening area of the lymphatic stomata (P< 0.01) and the concentration of absorbed trypan blue of the diaphragm (P<0.05). Sper/NO could not make a further change in the samples pretreated by nifedipine (P> 0.05). The results indicate that NO can decrease [Ca(2+)](i) in the RPMC through the NO-cGMP pathway. This procession is related with the L- type voltage-gated Ca(2+) channel. NO enlarges the opening area of the lymphatic stomata and enhances the lymph drainage of tracer by NO-cGMP-[Ca(2+)](i) pathway.

PCR products with heterozygous mutations containing two types of heteroduplexes / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To confirm that PCR products with heterozygous mutations contain not only wide-type and mutant homoduplexes, but also two types of heteroduplexes.</p><p><b>METHODS</b>An insertion-deletion mutation in the exon 1 of KRT9 gene (497delAinsGGCT), which caused Chinese epidermolytic palmoplantar keratoderma (EPPK) was investigated by polymerase chain reaction (PCR), polyacrylamide gel electrophoresis (PAGE) and denaturing high-performance liquid chromatography(DHPLC).</p><p><b>RESULTS</b>Two heteroduplexes and two homoduplexes in the PCR product from the heterozygous mutation of the exon 1 of KRT9 (497delAinsGGCT) were detected.</p><p><b>CONCLUSION</b>PCR products from KRT9 gene with heterozygous mutations contain two types of heteroduplexes. It is without the need to perform heating and cooling PCR products obtained from heterozygous mutations in advance before the mutation screening steps such as denaturing gradient gel electrophoresis (DGGE), temperature gradient gel electrophoresis (TGGE), conformation-sensitive gel electrophoresis (CSGE), DHPLC and heteroduplex analysis (HA), etc.</p>

Preliminary study on effects of Trichosanthes kirilowi root on hela cells / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the cytotoxic effect of extracts of Trichosanthes kirilowi (TK) root on Hela cells in vitro and its probable anti-tumor mechanism.</p><p><b>METHODS</b>The cytotoxic effect in vitro on the growth of Hela cells was evaluated by microculture tetrazolium assay (MTT). Cell ultrastructural changes were examined by scanning electron microscopy (SEM) and transmission electron microscopy (TEM), and DNA agarose electrophoresis was performed to determine apoptosis and biochemical changes of Hela cells.</p><p><b>RESULTS</b>Exposure of Hela cells to TK extracts for 24-48 hrs resulted in a cell growth arrest, which showed in a time- and dose-dependent manner (r > 0.880, P < 0.01). With SEM and TEM, marked changes were observed, including microvilli disappearance or reduction, cell membrane vesiculation, cell shrinkage, condensation of chromosomes and apoptotic bodies with complete membrane. Besides, the apoptosis of Hela cells was confirmed by typical DNA ladder formation on gel electrophoresis.</p><p><b>CONCLUSION</b>Extracts of TK has a marked anti-tumor activity and could induce apoptosis of Hela cells.</p>

The intervention effects of Angelica sinensis, Salvia miltiorrhiza and ligustrazine on peritoneal macrophages during peritoneal dialysis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of Angelica sinensis, Salvia miltiorrhiza and Ligustrazine on function of peritoneal macrophages during peritoneal dialysis.</p><p><b>METHODS</b>Peritoneal macrophages of mice were cultured in culture medium (control), peritoneal dialysate (PD), drugs contained PD containing Angelica, Salvia and Ligustrazine combined (PD-ASL) or separated (PD-A, PD-S, PD-L) with concentration of 2 micrograms/ml, 10 micrograms/ml and 100 micrograms/ml, separately for 24 hrs. The nitric oxide (NO) content, methyl thiazolyl tetrazolium (MTT) reducing capacity (MTT-RC) and phagocytosis capacity of macrophages were determined and compared.</p><p><b>RESULTS</b>NO content and MTT-RC of macrophages cultured in PD group were significantly lower than those of the control (P < 0.01), as compared with those in drug contained PD groups, the NO content in the PD-L group and the MTT-RC in the PD-ASL group were higher significantly (P < 0.01). The phagocytosis capacity and NO content in the PD-ASL group were raised along with the increased concentration of drug in PD.</p><p><b>CONCLUSION</b>Administering Chinese herbal medicine during peritoneal dialysis has important significance in improving the defense function of peritoneal macrophages, reducing the incidence of peritonitis and enhancing the therapeutic effect of peritoneal dialysis.</p>

Influence of peritoneal dialysate on nitric oxide production and viability of macrophage / 浙江大学学报·医学版

OBJECTIVE: To investigate the influence of commercial peritoneal dialysate (CDS) on function of macrophage. METHODS: Cultured peritoneal macrophages were divided into two experimental groups and their controls.(1) Macrophages were cultured in conditioned culture medium containing 50%CDS (0.139 mol/L glucose) for 24 h. (2)Macrophage were exposed to CDS containing 0.139mol/L glucose for 10, 30 and 60 min respectively, and then cultured in CDS-free medium for 24 h. The nitric oxide (NO) production and MTT in two groups were measured. In each control group, CDS was replaced by same amount of culture medium. RESULTS: NO production and MTT reduction ability (related to cell viability) of experimental groups were remarkably lower than those of controls and the NO production and MTT reduction in 60min CDS-exposed group were lower than that of 10 min and 30 min CDS-exposed group. CONCLUSION: Dialysate may have detrimental effects on viability and other function of macrophage and these effects may related to time length of CDS exposure.

Effects of nitric oxide on pleural lymphatic stomata and lymphatic drainage in the rat / 生理学报

To investigate the effects of nitric oxide (NO) on the pleural lymphatic stomata and lymph absorption from the pleural cavity, the NOS (nitric oxide synthase) inhibitor N(omega)-nitro-L-arginine-methyl-ester (L-NAME) and the NO donor isosorbide dinitrate (ISDN) were injected into the peritoneal cavity of the rats respectively. Trypan blue was used as a tracer. Then the concentrations of NO and trypan blue in the blood serum were measured, and the ultrastructural changes in pleural lymphatic stomata were observed under a scanning electron microscope (SEM) and studied by a computer image processing system attached to SEM. It turned out that the concentration of NO in the serum was 49.34+/-18.47 micromol/L, and the area and density of the pleural lymphatic stomata were 6.80+/-1.13 microm(2) and 170.24+/-66.60 /0.1 mm(2) respectively in the NO donor group. The concentration of NO reduced to 17.72+/-6.58 micromol/L, and the area and density of the pleural lymphatic stomata were 5.72+/-1.54 microm(2) and 61.71+/-12.73/0.1 mm(2) in the NOS inhibitor group. We found that the area and density of the pleural lymphatic stomata were positively correlated with the NO quantity. After the tracer was injected into the pleural cavity, the NO donor group exhibited a higher trypan blue concentration than the control group. The ability of the pleura to absorb trypan blue was enhanced because of the large opening of the stomata. It is suggested that NO can increase lymph absorption of the pleura by relaxing pleural lymphatic stomata.

A study of human enteric nervous system development in the middle stage of embryogenesis / 浙江大学学报·医学版

OBJECTIVE: To investigate the development of the enteric nervous system(ENS) in the middle stage(the 4th approximate, equals 6th months) of human embryogenesis. METHODS: A histologic evaluation of the clonic enteric nervous system was done using NADPHd histochemistry, PAP immunohistochemistry, with anti-PGP 9.5 and anti-S-100 protein. RESULTS: During this stage of embryology three things were noted. (1)The nerves in the myenteric layer increased markedly. This was shown by the PGP 9.5 immunoreactive nerves spreading out and the S-100 protein immunoreactive nerves forming a "bamboo basket"pattern. (2)The whole myenteric colon showed nitrigeric nerves paralleling the growth of the myofibers in the circular muscle layer. Nitrigeric perikara were rarely found in the extrinsic submucosal layer.(3) In the whole-mounted preparations reactive nerves formed the complex nerve net in the myenteric layer. CONCLUSION: The middle stage of embryogensis is very important to the development of the colon ENS.

A mouse model of severe acute pancreatitis induced by caerulein plus lipopolysaccharide / 中国医学科学院学报

<p><b>OBJECTIVE</b>To set up a convenient nontraumatic mouse model of severe acute pancreatitis(SAP).</p><p><b>METHODS</b>Mice received intraperitoneal injections with caerulein and lipopolysaccharide (LPS). Serum amylase and pancreatic moisture content were measured during experiment. The histo pathological changes of pancreas and relevant organs were observed under light microscope.</p><p><b>RESULTS</b>Serum amylase and pancreatic moisture content increased and pancreatic interstitial edema, inflammatory cellular infiltration, parenchymal necrosis as well as parenchymal hemorrhages were happened in the caerulein plus LPS group, and the lesions of other organs including stomach, ileum, spleen, and lung were seen as well. In the careulein group, there was only pancreatic interstitial edema with no parenchmal necrosis or hemorrhage, and the rest organs were normal.</p><p><b>CONCLUSIONS</b>The SAP mouse model induced by caerulein plus LPS has the same pathological characteristics of human SAP, which can be used for human SAP studies.</p>

Plasma adenomedullin, calcitonin gene-related peptide and c-type natriuretic peptide in rats with hypoxic pulmonary hypertension / 第三军医大学学报

Objective　To explore the effects of hypoxia on the syntheses and secretion of adrenomedullin (AM), calcitonin gene related peptide (CGRP) and c-type natriuretic peptide (CNP) and the relationship between these peptides. Methods　Rat models were established with hypoxia for 10, 20 and 30 d respectively and rats under normal altitude were served as control. Pulmonary artery pressure and the maximum increasing speed of right ventricle (RVdp/dtmax) were measured in every group. The dynamic changes of AM, CGRP and CNP concentrations in plasma were studied with radioimmunoassay. Results　During hypoxia, pulmonary artery pressure and RVdp/dtmax were enhanced. Plasma AM and CNP concentrations were increased while CGRP was decreased significantly. The plasma level of AM had positive correlation with that of CNP, but negatively correlated with that of CGRP. Conclusion　Results indicate that hypoxia may cause pulmonary artery pressure change and right ventricle has compensatory reaction to hypoxic pulmonary hypertension. Dynamic changes of plasma AM, CGRP and CNP concentrations can be regarded as indexes for condition of illness.

Plasma adenomedullin, calcitonin gene-related peptide and c-type natriuretic peptide in rats with hypoxic pulmonary hypertension / 第三军医大学学报

Objective　To explore the effects of hypoxia on the syntheses and secretion of adrenomedullin (AM), calcitonin gene related peptide (CGRP) and c-type natriuretic peptide (CNP) and the relationship between these peptides. Methods　Rat models were established with hypoxia for 10, 20 and 30 d respectively and rats under normal altitude were served as control. Pulmonary artery pressure and the maximum increasing speed of right ventricle (RVdp/dtmax) were measured in every group. The dynamic changes of AM, CGRP and CNP concentrations in plasma were studied with radioimmunoassay. Results　During hypoxia, pulmonary artery pressure and RVdp/dtmax were enhanced. Plasma AM and CNP concentrations were increased while CGRP was decreased significantly. The plasma level of AM had positive correlation with that of CNP, but negatively correlated with that of CGRP. Conclusion　Results indicate that hypoxia may cause pulmonary artery pressure change and right ventricle has compensatory reaction to hypoxic pulmonary hypertension. Dynamic changes of plasma AM, CGRP and CNP concentrations can be regarded as indexes for condition of illness.