The application of genetic engineering technology in modern agriculture shows its outstanding role in dealing with food shortage. Traditional medicinal plant cultivation and collection have also faced with challenges, such as lack of resources, deterioration of environment, germplasm of recession and a series of problems. Genetic engineering can be used to improve the disease resistance, insect resistance, herbicides resistant ability of medicinal plant, also can improve the medicinal plant yield and increase the content of active substances in medicinal plants. Thus, the potent biotechnology can play an important role in protection and large area planting of medicinal plants. In the development of medicinal plant genetic engineering, the safety of transgenic medicinal plants should also be paid attention to. A set of scientific safety evaluation and judgment standard which is suitable for transgenic medicinal plants should be established based on the recognition of the particularity of medicinal plants.
Genetic Engineering ; Plant Diseases ; genetics ; prevention & control ; Plants, Genetically Modified ; chemistry ; genetics ; growth & development ; metabolism ; Plants, Medicinal ; chemistry ; genetics ; growth & development ; metabolism

Objective To investigate the clinical efficacy of atlantoaxial pedicle screw fixation plus bony fusion in treatment of traumatic upper cervical instability.Methods From October 2009 to August 2013,29 patients with traumatic upper cervical spine instability were treated with posterior atlantoaxial pedicle screws.The patients underwent autografting (n =19) and allografting (n =10) for spinal fusion.Surrcal outcomes were recorded including intraopcrativc blood loss,operation time,with or without nerve,blood vessel and spinal cord injury,wound healing and bone fusion rate.Results All operations were completed smoothly with operation time of 110 minutes (range,85-135 minutes) and blood loss of 150 ml (range,80-500 ml).At the follow-up of 10 months to 5 years (mean 18 months),bony fusion was detected for all the patients.Postoperative radiographs verified all patients were bony fusion with satisfactory cervical spine stability.No complications of reduction loss,fixation failure,and spinal cord or vertebral artery injury were observed except for 1 patient with low viruleut infection and 2 with delayed wound healing.Conclusion Single posterior atlantoaxial pedicle screw fixation provides security and reliable stability in treatment of upper cervical instability,however wound healing problems should be taken seriously.

Objective To investigate the effect of curcumin an extract of a Chinese medical herb on the sensitivity of CD133 + rectal cancer cells to radiotherapy.Methods In vitro experiments:CD133 +cells were purified with immunomagnetic beads from HRT-18 cell line and divided into curcumin group,radiotherapy group and curcumin plus radiotherapy group.MTT assay and Annexin V/PI staining were used to measure the proliferation and apoptosis of the cells.In vivo experiments:Transplanted rectal tumor was established in 46 nude mice and randomly divided into curcumin group,radiotherapy group and curcumin plus radiotherapy group.Tumor size and apoptosis were detected by daily observation and TUNEL staining respectively.Results Curcumin inhibited proliferation and apoptosis of CD133 + rectal cancer cells when combined with radiotherapy.It also significantly increased the growth inhibition of rectal tumor and promoted the apoptosis of rectal cancer in vivo.MTT assay showed that after 24 hours,compared with that of radiotherapy group(14.6％ ± 1.0％),curcumin plus radiotherapy group (18.7％ ± 1.7％) inhibited the growth of the tumor(P ＜ 0.01).Annexin V/PI showed that curcumin plus radiotherapy group (28.8％ ±3.7％) was significantly different from the radiotherapy group(13.1％ ± 1.4％) in cell apoptosis (P ＜0.01).In vivo,after 6 days,tumor volume (521 ± 79) mm3 in curcumin plus radiotherapy group was significantly lower than that of radiotherapy group(717 ± 134) mm3 (P ＜ 0.01) ; TUNEL staining results indicated that the RCST in curcumin plus radiotherapy group (26.1％ ± 3.3％) were higher than that in radiotherapy group (12.0％ ± 2.1％) (P ＜ 0.01).Conclusions Curcumin significantly enhances the radiosensitizing effect for CD133 + rectal cancer cells.

Objective To investigate kupffer cells (KCs) expressing indoleamine 2,3-dioxygenase(IDO)in the inhibition of allogeneic T-cell proliferation in vitro. Methods Real-time PCR was used to investigate the expression of IDO mRNA and FasL mRNA in KCs pretreated with or without IFNγ. High performance liquid chromatography was used to analyze the catabolism of tryptophan by IDO from KCs. Allogeneic T-cell response was used to confirm the inhibition of KCs in vitro. The proliferation of lymphocytes was detected using [3 H] thymidine incorporation. Cell cycle and lymphocyte apoptosis were evaluated by flow cytometric assay. Results Real-time PCR revealed IDO mRNA and FasL mRNA expression in KCs pretreated with IFN-γ. IDO catabolic effect was confirmed by a decrease in tryptophan and increase in kynurenine concentration. KCs expressing IDO and FasL from BABL/c mice acquire the ability to suppress the proliferation of T-cells from C57BL/6, which could be blocked by the addition of 1-methyl-tryptophan and anti-FasL antibody. The co-cultured T-cells with KCs expressing IDO and FasL could induce allogeneic T-cell apoptosis and exhibited cell-cycle arrest in G1. Conclusion In addition to the Fas/FasL pathway, IDO may also play an important role in KCs to inhibit allogeneic T-cell proliferation in vitro.

Taxol is one of the most potent anti-cancer agents, which is extracted from the plants of Taxus species. Isopentenyl diphosphate isomerase (IPI) catalyzes the reversible transformation between IPP and DMAPP, both of which are the general 5-carbon precursors for taxol biosynthesis. In the present study, a new gene encoding IPI was cloned from Taxus media (namely TmIPI with the GenBank Accession Number KP970677) for the first time. The full-length cDNA of TmIPI was 1 232 bps encoding a polypeptide with 233 amino acids, in which the conserved domain Nudix was found. Bioinformatic analysis indicated that the sequence of TmIPI was highly similar to those of other plant IPI proteins, and the phylogenetic analysis showed that there were two clades of plant IPI proteins, including IPIs of angiosperm plants and IPIs of gymnosperm plants. TmIPI belonged to the clade of gymnosperm plant IPIs, and this was consistent with the fact that Taxus media is a plant species of gymnosperm. Southern blotting analysis demonstrated that there was a gene family of IPI in Taxus media. Finally, functional verification was applied to identify the function of TmIPI. The results showed that biosynthesis of β-carotenoid was enhanced by overexpressing TmIPI in the engineered E. coli strain, and this suggested that TmIPI might be a key gene involved in isoprenoid/terpenoid biosynthesis.
Amino Acid Sequence ; Carbon-Carbon Double Bond Isomerases ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; Paclitaxel ; biosynthesis ; Phylogeny ; Plant Proteins ; genetics ; Taxus ; enzymology ; genetics

[ ABSTRACT] AIM: To determine the aberrant methylation status in the gene promoter regions of CDH13, RASSF1A, DLEC1, SEPT9 and RUNX3 by detecting the plasma specimens and the value of their combined detection for di-agnosis of lung cancers.METHODS:Nest methylation specific PCR ( nMSP) was used to detect the promoter methylation status of the 5 genes in the plasma from 106 normal controls, lung cancer tissues, lung benign tissues and the plasma from 106 patients with lung cancers.Multiple displacement amplification ( MDA) was used to amplify modified genomic DNA to solve the problem of insufficient of plasma DNA template.RESULTS: The positive rates of promoter methylation of CDH13, RASSF1A, DLEC1, SEPT9 and RUNX3 in the lung cancer tissues were 51.9%, 44.3%, 54.7%, 36.8%, 24.5%, respectively, and those in the plasma were 46.2%, 41.5%, 50.9%, 31.1%, 19.8%, respectively.The re-sults of the Kappa consistency check showed that the lung cancer tissues and the plasma had obviously coherence in the methylation status of the 5 gene promoter regions.Combination of DLEC1, CDH13, RASSF1A, and SEPT9 had a higher di-agnostic efficiency than the others, as their ACC value was 0.8208 and youden index was 0.6415 ( with the sensitivity of 81.13% and the specificity of 83.02%) .CONCLUSION:Combination detection of promoter methylation of lung cancer-related genes in the plasma is expected to apply to the early diagnosis of lung cancer.

Objective Preterm birth is a live birth delivered before 37 weeks of gestation.Preterm birth rates have risen in recent years,and preterm birth is the leading cause of perinatal morbidity and mortali-ty worldwide.Both environmental and genetic factors likely play important roles in the development of pre-term birth.MicroRNAs(miRNAs)are 18 to 25 nucleotides,single stranded non-coding RNAs that regulate a wide range of biological processes in development and human disease.In this study,we have investigated the differential expression of hsa-miR-28-3p and ADAM12 in cord blood mononuclear cells of preterm birth and term birth.Methods The cord bloods were collected from the patients of Putuo District Institute of Materni-ty and Child Health.The preterm group(30 patients)was the patients with spontaneous preterm delivery,and the control group(30 patients)was the patients delivered normal infants at term.The expression levels of hsa-miR-28-3p and ADAM12 mRNA were directly generated by real-time PCR.Results The gestational age and birth weight of preterm group was(30.92 ±1.73)weeks and(1 646 ±357)g,and the control group was (39.73 ±0.58)weeks and(3 301 ±394)g.The hsa-miR-28-3p expression of preterm group(1.03 ±0.23) was significantly lower than that of control group(2.32 ±0.52)in the peripheral blood mononuclear cells (P <0.01 );the ADAM12 mRNA expression of preterm group(0.037 8 ±0.005 6)was significantly higher than that of control group(0.027 6 ±0.003 9)(P <0.05);the hsa-miR-28-3p expression was significantly correlated with the ADAM12 mRNA expression(r =-0.634 1 ,P <0.01 ).Conclusion The lower hsa-miR-28-3p expression of cord blood mononuclear cells may up-regulate ADAM12 mRNA expression,and promotes the occurrence of spontaneous preterm birth.

OBJECTIVETo explore the expression of phospholipase C-gamma 1 (PLC-gamma1) alternative splicing variants in rats.

METHODSAccording to the sequence of human PLCG1 splicing variant, specific primers for rat PLC-gamma1 were designed and synthesized. The rat RNA was reverse transcribed into cDNA, which was amplified using the specific primers, and the PCR products were sequenced and analyzed using BLAST and bioinformatics methods. Totally 21 rat tissue samples were examined, including the heart, liver, lung, kidney, eyeball, and brain obtained in 3 different embryonic stages, 7 different early postnatal stages, and in adulthood.

RESULTSThe result did not show that rat PLC-gamma1 had the same splicing variant (PLC-gamma1a, NM_002660) as human does.

CONCLUSIONSThe same splicing variant of PLC-gamma1 detectable in human may not exist in rats, and the pre-mRNA may undergo splicing resulting predominantly in PLC-gamma1b mRNA. Very likely, the alternative splicing site of rat PLC-gamma1 is not identical to that of human.

Alternative Splicing ; Animals ; Base Sequence ; Molecular Sequence Data ; Phospholipase C gamma ; genetics ; RNA Precursors ; genetics ; RNA Splice Sites ; genetics ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVETo investigate the effects of histamine on the neurotoxicity induced by beta-amyloid(1-42)(Abeta42) in rat phaeochromocytoma (PC12) cells.

METHODSThe in vitro model of Alzheimer's disease was constructed with A beta42-treated PC12 cells. Cell morphology and MTT assay were used to evaluate the cell toxicity and histamine effects. The different histamine antagonists were applied to investigate the involvement of receptor subtypes.

RESULTThe neurotoxicity was induced by A beta42 in a concentration-dependent manner, which was reversed by histamine at concentration of 10(-7), 10(-6) mol/L. The effect was reversed by H(2) antagonist zolantidine and H(3)antagonist clobenpropit, but not by H(1) antagonist diphenhydramine.

CONCLUSIONHistamine reduces neurotoxicity induced by beta-amyloid(1-42), which may be mediated by H(2) and H(3)receptors.

Alzheimer Disease ; chemically induced ; metabolism ; prevention & control ; Amyloid beta-Peptides ; toxicity ; Animals ; Benzothiazoles ; pharmacology ; Diphenhydramine ; pharmacology ; Dose-Response Relationship, Drug ; Histamine ; pharmacology ; Histamine H2 Antagonists ; pharmacology ; Histamine H3 Antagonists ; pharmacology ; Imidazoles ; pharmacology ; Neuroprotective Agents ; metabolism ; pharmacology ; PC12 Cells ; Phenoxypropanolamines ; pharmacology ; Piperidines ; pharmacology ; Rats ; Receptors, Histamine H2 ; metabolism ; Receptors, Histamine H3 ; metabolism ; Thiourea ; analogs & derivatives ; pharmacology

OBJECTIVETo observe the effect of Yiqi Huoxue Qingre Huashi Recipe (YHQHR, a recipe capable of supplementing qi, activating blood, clearing heat, and dissipating dampness) on ulcer healing and Helicobacter pylori (Hp) eradication rate in Hp positive peptic ulcer patients, and to explore coccoid Hp occurrence in the eradication.

METHODSTotally 80 Hp positive peptic ulcer patients were assigned to the treatment group and the control groups by random digit table, 40 in each group. All patients received standard triple therapy of Western medicine for 2 successive weeks. Those in the control group additionally took omeprazole enteric coated tablet, 20 mg each time, once per day for 4 successive weeks. Those in the treatment group additionally took YHQHR, twice per day for 6 successive weeks. The ulcer healing was observed and recorded by gastroscope after discontinued medication of 14 days. The effective rate of ulcer healing under endoscope was statistically calculated. Rapid urease test (RUT) was performed in one small piece of tissue from corpora ventriculi and sinuses ventriculi using 14C breathe test (UBT). Gastric juice was collected from the stomach. Hp urease gene amplification test (urea A-PCR) was performed in living tissue from gastric antrum. Results obtained from the above three test methods were recorded and assessed to decide the final eradiation rate. Gastric mucosa tissue was observed under electron microscope,attempting to find non-eradicated Hp, which was further observed.

RESULTSThe total curative effect under gastroscope was 97.5% (39/40 cases) in the treatment group, obviously higher than that in the control group (80.0%, 32/40 cases) (P < 0.05). The eradication rate of Hp was 75.0% (30/40 cases), obviously better than that of the control group (52.5%, 21/40 cases) (P < 0.05). The total positive Hp numbers after treatment was 14C UBT (12), RUT (8), and urea A-PCR (27), respectively. The Hp positive rate detected by 14C UBT and RUT was lower than the Hp positive rate detected by urea A-PCR (P < 0.05). Rod-like and coccoid Hp bacteria could be observed under electron microscope.

CONCLUSIONYHQHR combined standard triple therapy was more effective than standard triple therapy alone in promoting ulcer healing and elevating the eradication rate of Hp.

Breath Tests ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; Helicobacter Infections ; drug therapy ; Helicobacter pylori ; Humans ; Omeprazole ; Peptic Ulcer ; drug therapy ; microbiology ; Urea