To providing evidence about nitrogen adequate application of Platycodon grandiflorum, the pot culture experiment was conducted to study the effect of nitrogen on the growth, physiological metabolism and the quality of P. grandiflorum. The activity of NR, GS and SOD, POD and CAT were determined. And the nitrate and ammonium nitrogen content, photosynthetic characteristics, active components of P. grandiflorum were determined. The results showed that the nitrate nitrogen content and P. biomass reached its maximum value, when NH4(+)-N/NO3(-) -N was 0: 100, the activity of NR. The activity of GS was the highest at the NH4(+) -N/NO3(-) -N ratio of 25:75 and ammonium nitrogen content was the highest at 75:25. The activity of SOD decreased and then increased with the increasing of NO3(-) -N. At the NH4(+) -N/NO3(-) -N ratio of 25: 75, the activity of CAT had its maximum value and the content of MDA had the minimum value. At the same time, the content of platycodon D was the highest at this treatment. The studies had shown that different nitrogen forms and ratio had a significant effect on the characteristics of photosynthetic physiology, nitrogen metabolism and resistance adjustment, growth and the quality of P. grandiflorum. The NH4(+) -N/NO3(-) -N ratio of 25: 75 was a suitable ratio of nitrogen forms for the growth of P. Grandiflorum and accumulating the content of platycodon D.
Ammonium Compounds ; metabolism ; Biomass ; Drugs, Chinese Herbal ; analysis ; metabolism ; Nitrates ; metabolism ; Photosynthesis ; Plant Leaves ; chemistry ; growth & development ; metabolism ; Platycodon ; chemistry ; growth & development ; metabolism

OBJECTIVESTo study the effect of Houxue Zhitong decoration on the expression of the mitochondria-initiated apoptosis pathway and transmembrane protein I an II of the epidural scar tissue.

METHODSA total of 60 New Zealand rabbits (weight: 2.5-3.0 kg) were randomly divided into four groups, sham operation group (D, n=15), control group (B, n=14), sodium hyaluronate group (C, n=15), Houxue Zhitong decoration group (D, n=15). Except for group A, 1.0 cm x 1.0 cm dura mater uncovered area laminectomy was performed at I (4) and I(5), covered with 0.5 ml sodium hyaluronate in group C, covered with same amount of saline in group B and D. First 2 weeks after operation, animals in group D were lavaged with 2.5 ml/kg Houxue Zhitong decoction by one a day for 14 days. Five rabbits of each group selected randomly were killed in the 2,4,8 weeks after laminectomy. The specimens were prepared for determination of the expression of Fas and FasL, at scar tissue by semiquantitative reserve transcription-polymerase chain reaction (RT-PCR). The degree of scar adhesion was evaluated according by Rydell method.

RESULTSThe adhesion area in group B was larger than of group C and D in the 4th and 8th week. However, the number of fibroblasts and inflammantory cells in group D was the least among the three groups in the 8th week. At 2, 4, 8 weeks after operation, as compared with group B the expression of Fas, FasL of group C and D were decreased (P < 0.05). Especially, at 2 weeks, as compared with group B the expression of this two cytokines of group D was significant decreased (P < 0.05), too. In group C and D the duramater adhesion was decreased (P < 0.05). The proliferation of fibroblast and fibroblastic function were inhibited (P < 0.05).

CONCLUSIONHuoxue Zhitong is able to down-regulated the expression of Fas, FasL, which inhibited the proliferation of fibroblast, the fibroblastic function and the synthesis of extracellular matrix in the epidural scar tissue. It is an effective way of reducing peridural scar formation and prevent the failed back surgery syndrome.

Animals ; Apoptosis ; drug effects ; Cicatrix ; genetics ; metabolism ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Epidural Space ; pathology ; Fas Ligand Protein ; genetics ; metabolism ; Female ; Gene Expression Regulation ; drug effects ; Rabbits ; Signal Transduction ; drug effects ; fas Receptor ; genetics ; metabolism

OBJECTIVETo study the effect of Huoxuezhitong recipe on the pathway of mitochondrial apoptosis and the expression of related gene (Caspase-3, -9, Bax, Bcl-2) in epidural scar tissue.

METHODSA total of 60 New Zealand rabbits, ranging in weight from 2.5 to 3.0 kg, were randomly divided into sham operative group (group A), control group (group B), sodium hyaluronate group (group C), Huoxuezhitong recipe group (group D, included Danggui 20 g, Chishao 20 g, Honghua 20 g, Ruxiang 15 g, Myrrh 15 g etc.) with 15 rabbits in each group. Except for group A, L4,5 vertebral plate were resected leading to 1.0 cm x 1.0 cm dura mater exposed area, which were covered with 0.5 ml sodium hyaluronate in group C, covered with same amount of saline in group B and D. At 2 weeks after operation, the rabbits of group D were administered with Huoxuezhitong recipe (2.5 ml/kg, once a day, continuous 14 d). Five rabbits of each group were selected randomly and were killed at 2, 4 and 8 weeks after operation; and the expression of Caspase-3, -9, Bax, Bcl-2 in scar tissue were surveyed by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe results by RT-PCR showed significant different in expressions of Caspase-3, -9, Bax, Bcl-2 of group A and others groups at each period (P < 0.01). At 2, 4, 8 weeks after operation, compared with group B, the expressions of Caspase-3, -9, Bcl-2 in group C and D were decreased (P < 0.05), but the expression of Bax in group C and D were increased (P < 0.05). At 8 weeks after operation, the expressions of Caspase-3, -9, Bax, Bcl-2 in group D was significant decreased compared with group C (P < 0.05).

CONCLUSIONHuoxuezhitong recipe can regulate the levels of expression of Caspase-3, -9, Bax, Bcl-2 mRNA and further induce the pathway of mitochondrial apoptosis, thereby, achieve the effect of prophylaxis for the proliferation and conglutination of epidural scar tissue.

Actins ; genetics ; metabolism ; Animals ; Apoptosis ; drug effects ; genetics ; Carthamus tinctorius ; Caspase 3 ; genetics ; metabolism ; Caspase 9 ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Genes, bcl-2 ; drug effects ; genetics ; Mitochondria ; drug effects ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; bcl-2-Associated X Protein ; genetics ; metabolism

This study was purposed to investigate the effect of phycocyanin at different concentration on proliferation of K562 cells, to detect the changes of integrin beta1 expression and intracellular focal adhesion kinase (FAK) gene expression on the surface K562 cells treated with phycocyanin, and to explore the possible mechanism of integrin beta1 effect on phycocyanin inhibiting proliferation of K562 cells. The expression level of integrin beta1 on the surface of K562 cells was evaluated by flow cytometry (FCM); the growth of K562 cells treated with phycocyanin was measured by MTT assay; the expression level of FAK mRNA was analyzed by relatively quantitative RT-PCR after four-day culture of K562 cells with phycocyanin of 40 microg/ml, 80 microg/ml and 160 microg/ml, respectively. The results showed that integrin beta1 expression on the surface of K562 cells was significantly higher than that in bone marrow mononuclear cells (BMMNC) from normal subjects. Phycocyanin could not change the level of integrin beta1 expression. Phycocyanin could increase the expression of FAK gene on K562 cells and inhibit the proliferation of K562 cells. It is concluded that phycocyanin can inhibit the proliferation of K562 cells through enhancing the conjunction of cell stroma with integrin beta1 on K562 cell surface, up-regulating the expression level of FAK gene in K562 cells, restoring the signaling pathway of proliferation inhibition mediated by integrin beta1. The possible mechanism of phycocyanin in the proliferation inhibition of K562 cells is to increase the expression of FAK gene. The phycocyanin may be considered as a potential agent for inhibition of cancer cell proliferation.
Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Proliferation ; drug effects ; Focal Adhesion Kinase 1 ; biosynthesis ; genetics ; Humans ; Integrin beta1 ; biosynthesis ; genetics ; K562 Cells ; Phycocyanin ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVETo investigate the expression and mutation of Mad1, Mxi1 and Rox genes in leukemia cells.

METHODSExpression and mutation of Mad1, Mxi1 and Rox genes in bone marrow mononuclear cells (BMMNC) from 26 de novo acute leukemia (AL) patients, and in peripheral blood mononuclear cells (PBMNC) from 30 healthy volunteers, as well as in 7 human leukemic cell lines were analyzed by reverse transcription-polymerase chain reaction (RT-PCR), single strand conformational polymorphism (SSCP) and DNA sequencing.

RESULTSRT-PCR showed that all the above cells expressed Mad1, Mxi1 and Rox mRNA. SSCP revealed four polymorphisms: two in Mad1, one each in Mxi1 and Rox. DNA sequencing detected nine missense mutations: two in Mad1 in AL patients, four in Mxi1 (three in AL patients and one in KG-1 cell line), and three in Rox in AL patients. The mutations of Mad1, Mxi1 and Rox mRNA were detected in 2, 3 and 3 patients, respectively.

CONCLUSIONIt is for the first time to demonstrate the mutations of Mad1, Mxi1 and Rox genes in AL patients suggesting these mutated genes involve in the pathogenesis of leukemia.

Adolescent ; Adult ; Aged ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; genetics ; metabolism ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; metabolism ; Cell Cycle Proteins ; genetics ; metabolism ; Female ; Humans ; Leukemia ; genetics ; metabolism ; Male ; Middle Aged ; Mutation ; Nuclear Proteins ; genetics ; metabolism ; Polymorphism, Single-Stranded Conformational ; Repressor Proteins ; genetics ; metabolism ; Tumor Suppressor Proteins ; genetics ; metabolism

To study insulino-mimetic effects of bis(alpha-furancarboxylato) oxovanadium (IV) (BFOV), a orally active antidiabetic vanadyl complex, on glucose uptake and lipogenesis in isolated rat adipocytes were determined by using 2-deoxy-D-[3H]-glucose and D-[3H]-glucose, respectively. Lipolysis was assayed by free fatty acids (FFA) released from isolated rat adipocytes treated with epinephrine. The results showed that BFOV, similar to insulin, concentration-dependently significantly enhanced the uptake of 2-deoxy-D-[3H]-glucose and the transformation from D-[3H]-glucose to lipid in isolated rat adipocytes, with the EC50 values of (0.31 +/- 0.08) mmol L(-1) and (0.49 +/- 0.12) mmol L(-1), respectively. Moreover, BFOV markedly inhibited FFA release from isolated rat adipocytes treated with epinephrine, and the IC50 value was (0.30 +/- 0.20) mmol L(-1). BFOV had insulino-mimetic effects such as enhancing glucose uptake and lipogenesis, as well as inhibiting lipolysis.
Adipocytes ; drug effects ; metabolism ; Animals ; Blood Glucose ; drug effects ; Hypoglycemic Agents ; pharmacology ; Insulin ; pharmacology ; Lipogenesis ; drug effects ; Male ; Organometallic Compounds ; pharmacology ; Rats ; Rats, Sprague-Dawley

Limosilactobacillus reuteri is a microbe intricately linked to humans and animal health.A thor-ough assessment of its safety and potential benefits is imperative prior to its application in human and ani-mals.In this investigation,we performed a comprehensive analysis encompassing genome sequencing,genomic analysis,and phenotypic characterization of L.reuteri Q35,an exceptionally proficient producer of reuterin.The whole genome sequencing results showed that the complete genome sequence spans 2145158 bp with a GC content of 38.9％ and encompasses 2121 genes.Initial identification of antibiotic-re-sistant genes,virulence factors,and toxin-coding genes in the genome substantiated the strain' s low-risk status.Subsequent tests for antibiotic resistance,acute oral toxicology,and hemolysis further confirmed its elevated safety level.The genome of L.reuteri Q35 was found to contain genes associated with adhe-sion and stress tolerance.Following exposure to artificial gastric juice and bile salt,the strain exhibited a higher survival rate and demonstrated a strong scavenging ability for hydroxyl free radicals in antioxidant capacity tests.These findings suggested that L.reuteri Q35 possesses unique probiotic properties.Addi-tionally,the genome of strain Q35 harbors three truncated oxaloyl-CoA decarboxylase genes (oxc1,oxc2 and oxc3),overexpression of which resulted in a significant increase in ammonium oxalate degradation from 29.5％ to 48.8％.These findings highlight that L.reuteri Q35 exhibits both favorable safety charac-teristics alongside beneficial properties,making it a promising candidate for treating metabolic disorders such as hyperoxaluria.

Objective To observe the effect of electrical stimulation combined with bladder function training on autonomic bladder pa-tients after S2-4spinal cord injury or cauda equina and pelvic nerve injury.Methods From January,2016 to August,2017,76 patients with au-tonomic bladder after spinal cord injury were divided into control group(n=29)and observation groups(n=47)in real world study.The con-trol group received routine bladder function training,and the observation group received bladder muscle stimulation therapy in addition.The micturition diary and urodynamic indexes were assessed before and three months after treatment.Results After treatment,the frequency of urinary incontinence and the number of micturition in 24 hours,and residual urine volume significantly reduced(t>3.251,P<0.01),the urine output every time significantly increased (t>5.000, P<0.001), the maximum bladder capacity and urine flow rate increased (t>1.922, P<0.05),and the detrusor pressure in the end of filling period significantly reduced(t>8.302,P<0.001).All the indexes above were better in the observation group than in the control group(t>1.919,P<0.05). Conclusion Electrical muscular stimulation combined with the bladder func-tional training could improve the urination function in the patients with autonomic bladder after spinal cord injury.

Previous studies demonstrated that interleukin-12 (IL-12) enhances the non-MHC-restricted cytotoxic activity of NK cells and facilitate specific allogeneic human cytotoxic T lymphocyte responses against fresh leukemia cells and cell lines. The Wilms' tumor gene, WT1 mRNA, has been used as a marker of minimal residual disease (MRD) for evaluating therapeutic efficacy of patients with leukemia or myelodysplastic syndrome (MDS). This study was aimed to investigate whether in vitro IL-12 can lower WT1 gene expression in peripheral blood monuclear cells (PBMNC) from patients with leukemia or MDS. PBMNC from these 30 patients and 5 healthy volunteers were cultured at 5 x 10(5) cells/ml alone with or without 100 units/ml of IL-12 for 3 days. WT1 mRNA was measured by competitive reverse transcription polymerase chain reaction (RT-PCR) since WT1 mRNA is considered as a marker of minimal residual disease (MRD) in leukemia and MDS. The results demonstrated that WT1 mRNA in PBMNC of 5 healthy volunteers was less than 10(3) copies/microg of total RNA. Following the 3-day IL-12 treatment, mean WT1 mRNA of PBMNC was reduced from 10(4.8) to 10(4.2) copies/microg of total RNA in 6 CML patients, from 10(5.4) to 10(4.8) copies/microg in 12 MDS patients and from 10(5.0) to 10(4.2) copies/microg in 5 AML patients in CR, but not reduced in 5 of 7 AML in non-CR. It is concluded that IL-12 significantly decrease the quantity of leukemia cells in PBMNC of most patients with MDS, CML and AML in CR. IL-12 may be of considerable benefit in the elimination of MRD in patients with hematological malignancies.
Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Interleukin-12 ; pharmacology ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; metabolism ; Neoplasm, Residual ; genetics ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; WT1 Proteins ; biosynthesis ; genetics

Neuropeptide Y (NPY), a sympathetic neurotransmitter, is highly associated with baroreflex dysfunction and multiple cardiac diseases such as diabetic myocardiopathy. In the present study, we aimed to explore the role of peripheral NPY Y1 receptor (Y1R) and Y2 receptor (Y2R), which are dominantly present in peripheral cardiovascular control, in baroreflex sensitivity (BRS) of streptozotocin (STZ)-induced diabetic rats. Peripheral Y1R and Y2R were antagonized by specific antagonists (BIBP 3226 and BIIE 0246, respectively) from subcutaneously implanted ALZET mini-osmotic pump in STZ-induced diabetic rats for 4 weeks. Then baseline systolic blood pressure, heart rate, cardiac function, BRS, plasma NPY and lipid levels were evaluated. We found that STZ led to increased plasma NPY and lipid level. And the STZ-increased lipid levels were reduced by BIBP 3226 and BIIE 0246. BIBP 3226 ameliorated the aberrant BRS, but had little effect on the impaired cardiac function of the STZ rats. BIIE 0246 alleviated sodium nitroprusside (SNP)-induced but not phenylephrine (PE)-induced aberrant baroreflex control of heart rate in the STZ rats. In addition, BIIE 0246 alleviated the bradycardia, but further impaired cardiac contractility in the STZ rats. These results suggest that peripheral Y1R and Y2R play different roles in STZ-induced impairment of BRS.
Animals ; Arginine ; analogs & derivatives ; pharmacology ; Baroreflex ; Benzazepines ; pharmacology ; Blood Pressure ; Bradycardia ; Diabetes Mellitus, Experimental ; drug therapy ; physiopathology ; Heart Rate ; Myocardial Contraction ; Neuropeptide Y ; blood ; Rats ; Receptors, Neuropeptide Y ; antagonists & inhibitors ; Streptozocin