Gene targeted mutagenesis and replacement can be used to modify gene sequence in genomic background without position effect or insertion inactivation in transgenic plants. Targeted mutagenesis organism has little biosafety concerns free of transgenes or marker genes. Gene targeted mutagenesis and replacement in high plants now appears to be a potential tool for gene functional analysis in situ, crops genetic improvement and molecular design. Zinc finger nuclease(ZFN)is most important and would be widely used in gene targeted mutagenesis and replacement through introducing double-strand breaks in genome. Strategies for gene targeted mutagenesis and replacement in plants is discussed. ZFN is described in detail from its structure, operation model and application in plants. Developmental prospect of ZFN in plant gene targeted mutagenesis and replacement is also discussed.

The aim of the present study was to investigate the effect of "nourishing liver and kidney" acupuncture therapy on motor and cognitive deficits,and the underlying mechanism following cerebral ischemia-reperfusion (I/R) via increasing the expression of brain derived neurotrophic factor (BDNF) and synaptophysin (SYN) in the hippocampus.Healthy adult male SD rats were randomly divided into sham operation group (n=51),model group (n=51),acupuncture group (n=51) and acupuncture control group (n=51).The middle cerebral I/R model was established.Acupunctures were performed in the acupuncture group and acupuncture control group at acupoints of Taixi (K103),Taichong (ST09) of both sides,for 30 min once daily every morning.The animals in the sham operation group and model group were conventionally fed in the cage,without any intervention therapy.The rats of each group were assessed with modified neurological severity scores (mNSS).The expression of BDNF and SYN in the hippocampus was detected by immunohistochemical SP method and the synaptic structure in hippocampus area was assessed morphologically and quantitatively at the 3rd,7th and 14th day.The Morris water Maze (MWM) test was used to evaluate the rats' learning and memory abilities on the 15th day after acupuncture.The animals in the acupuncture control group and sham operation group presented no neurological deficit.In the acupuncture group,the nerve functional recovery was significantly better than that in the model group at the 7th and 14th day after modeling.The average MWM escape latency in the acupuncture group was shorter than that in the model group at the 3rd,4th and 5th day.The number of crossings of the platform quadrant in the acupuncture group was significantly more than that in the model group.At the each time point,the expression levels of BDNF and SYN in the hippocampal regions increased significantly in the model group as compared with the sham operation group and the acupuncture control group.In the acupuncture group,the expression levels of BDNF at the 7th and 14th day increased more significantly than those in the model group.In the acupuncture group,the expression levels of SYN at the each time point increased more significantly than those in the model group.The post-synaptic density (PSD) was significantly increased and the synapse cleft width was narrowed in the acupuncture group as compared with other groups.The synaptic curvatures were improved obviously in the acupuncture group in contrast to the model group.It was concluded that the "nourishing liver and kidney" acupuncture therapy has positive effects on behavioral recovery,as well as learning and memory abilities,probably by promoting the expression of BDNF and SYN,and synaptic structure reconstruction in the ipsilateral hippocampus after I/R in rats.The "nourishing liver and kidney" acupuncture therapy can promote the functional recovery in rats after cerebral ischemia injury.

GJ-4 is crocin enrichments extracted from Gardenia jasminoides J. Ellis, and our previous studies have shown that GJ-4 significantly improved learning and memory impairment induced by Aβ in mice. Herein, a memory deficit model was developed by injecting okadaic acid (OA) into the lateral ventricle of mice, and the neuroprotection and underlying mechanism of GJ-4 on neuronal injury caused by Tau hyperphosphorylation were investigated. The Animal Care & Welfare Committee, Institute of Materia Medica, CAMS & PUMC has approved all procedures (No.00000318). GJ-4 at different doses was intragastric administration to mice for 16 days. Step-down test and Morris water maze test showed that GJ-4 could significantly improve OA-induced memory impairment in mice, and reduced the loss of Nissl bodies in the hippocampus of mice. GJ-4 could also decrease the phosphorylation level of Tau protein at Ser396, Thr231 and Ser404 via increasing protein phosphatase 2A (PP2A) activity and inhibiting glycogen synthase kinase-3β (GSK-3β) activity. Besides, further researches indicated that GJ-4 could inhibit the level of oxidative stress in the brain of OA mice, reduce neuronal apoptosis and inhibit the neuroinflammation mediated by activation of astrocytes in the hippocampus of mice, and eventually achieve its effects in improving learning and memory impairment in mice. According to these findings, we anticipated that GJ-4 might be a potential therapeutic drug for Alzheimer's disease.

OBJECTIVETo explore the underwater decompression schedule for 100 m Trimix conventional diving operations and evaluate its safety through a simulated rabbits Trimix conventional diving.

METHODSAccording to the Haldane theory, the assumed time units, the classification of tissue compartments, the nitrogen super-saturation safety coefficient and the selection of methods used for the calculation of the simulated 100 m Trimix conventional diving schedule were properly selected, and the calculating method for the dive decompression schedule was thus firmly established. In our experiments, five tissue compartments were selected during the calculation of decompression schedule: 5 min, 10 min, 20 min, 40 min and 75 min, and the nitrogen super-saturation safety coefficient was calculated by 1.6. Eight New Zealand rabbits were performed a simulated 100 m Trimix dive program which was established according to the Haldane theory, and eight rabbits for intact group. The tissues wet/dry ratio and ethology were detected and observed before and after the simulated diving to evaluate the safety of decompression schedule.

RESULTSBy using the developed underwater decompression schedule, abnormal ethology changes in rabbits could not be observed after compression and decompression to the surface; and the tissues wet/dry ratio of simulated diving rabbits had no significant changes compared with the intact group (P > 0.05).

CONCLUSIONThe decompression schedule calculated by Haldane theory seemed to be safe and reliable, the diving breathing gas concentration did not cause oxygen toxicity and nitrogen narcosis among the dive rabbits, and dive efficiency was greatly improved by using enriched oxygen gas in UPTD safety range during decompression.

Animal Experimentation ; Animals ; Decompression ; Diving ; Helium ; Nitrogen ; Oxygen ; Rabbits

Humans ; Immunotherapy ; methods ; Lung Neoplasms ; immunology ; therapy ; Vaccines ; therapeutic use

The primary aim of this research is to systematically sort out and analyze available clinical documents for chronic congestive heart failure in traditional Chinese medicine (TCM) and integrated traditional Chinese and Western medicine, and to explore the diagnosis and treatment rule of TCM syndrome factors with data mining method.

OBJECTIVETo investigate allelic frequencies of interluekin-10 (IL-10) gene promoter in Miao, Dong and Buyi ethnics of Guizhou.

METHODSTaqMan MGB-based real-time PCR was used to determine the genotypes of IL-10 -819 and IL-10 -592 in 589 Miao, Dong and Buyi ethnics of Guizhou.

RESULTSThe allelic frequency of IL-10 -819 in Miao ethnics was significantly different from those in Dong or Buyi ethnics. Allelic frequencies of IL-10 -592 in Miao ethnics was significantly different from those in Dong or Buyi ethnics. In Miao, Dong and Buyi ethnics, the distributions of genotype frequencies of IL-10 -819 and IL-10 -592 were statistically different from Han ethnics from Guizhou and Taiwan of China as well as South Koreans.

CONCLUSIONThere is a heterogeneity in the frequencies of polymorphisms of IL-10 promoter among different ethnic groups.

Alleles ; Asian Continental Ancestry Group ; ethnology ; genetics ; China ; ethnology ; Gene Frequency ; Genetics, Population ; Genotype ; Humans ; Interleukin-10 ; genetics ; Polymorphism, Single Nucleotide ; Population Groups ; genetics ; Promoter Regions, Genetic

Objective To investigate the effect of acupuncture plus rehabilitation training on upper limb functional recovery in convalescent stroke patients.Methods One hundred and three convalescent stroke patients with upper limb motor dysfunction were randomized to a treatment group (54 cases) and a control groups (49 cases). Both group received conventional rehabilitation training. In addition, the treatment group received along-meridian syndrome differentiation-based acupuncture and the control group, sham acupuncture. Upper limb Fugl-Meyer Assessment (U-FMA) score, the Wolf Motor Function Test (WMFT) score, the modified Barthel Index (MBI) score and the 36-Item Short Form Health Survey (SF-36) score were recorded in the two groups of patients before and at the end of treatment and at the 3-month follow-up.Results In the two groups there were statistically significant differences in the U-FMA score, the WMFT score and the MBI score at the end of treatment and the 3-month follow-up compared with before (P>0.05 orP<0.01). At the end of treatment and the 3-month follow-up there were statistically significant differences in the U-FMA score, the WMFT score and the MBI score between the treatment and control groups (P<0.05 orP<0.01). The scores of SF-36 items except social functioning and bodily pain had statistically significant differences within the two groups at the end of treatment and the 3-month follow-up compared with before and between the two groups at the end of treatment and the 3-month follow-up (P<0.01 orP<0.05).Conclusion Acupuncture plus rehabilitation training can markedly improve upper limb function in stroke patients.

Objective To observe the correlation between conventional coagulation tests and Dabigatran concentration in order to find proper test method to predict the bleeding risk of patients receiving Dabigatran.Methods The clinical data of forty-nine non-valvular atrial fibrillation (NVAF) patients who took Dabigatran in Peking University Third Hospital from 2015 to 2017 were analyzed and the bleeding rates were calculated.The plasma samples from twenty healthy volunteers were collected and mixed up into normal pooled plasma (NPP).Different doses of active Dabigatran were mixed with NPP,making Dabigatran concentrations from 0 to 1 000 ng/mL.Prothrombin time (PT),activated partial thromboplastin time (APTT),thrombin time (TT),fibrinogen (Fib) and diluted Russell viper venom time (dRVVT) were determined.Fresh whole blood samples from three normal volunteers were collected and mixed with Dabigatran in the same way to perform thromboelastogram (TEG).Results The total bleeding rate of NVAF patients receiving Dabigatran was 28.6％,among which gastrointestinal bleeding accounts for the most.APTT,dRVVT,R time (R) and clot index (CI) showed a good linear correlation with Dabigatran concentration.The sensitivity of dRVVT was the best,followed by APTT.R and CI showed low sensitivity compared with APTT and dRVVT.Conclusion APTT and dRVVT should be good parameters for monitoring the bleeding risk of Dabigatran and helpful for physicians to choose proper point-in-time for withdrawal of Dabigatran and reduce bleeding events.

OBJECTIVETo study the frequency of a 9 bp deletion polymorphism of mitochondrial DNA (mtDNA) in ethnic Miao, Buyi and Dong populations from Guizhou province.

METHODSPolymerase chain reaction-polyacrylamide gel electrophoresis (PCR-PAGE) was used to detect the 9 bp deletion. The result was verified with DNA sequencing.

RESULTSTwo polymorphisms, including a standard pattern and a short pattern (the 9 bp deletion), were found among the three ethnic groups. The frequency of short pattern in 304 males was 23.0%. Respectively, those of Miao, Buyi and Dong ethnics were 28.6%, 26.8% and 13.7%. A statistically significant difference was detected among the three groups (P<0.05).

CONCLUSIONThe frequencies of the 9 bp polymorphism were relatively high among ethnic Miao, Buyi and Dong populations from Guizhou, and there was a significant difference between the three.

Base Sequence ; China ; ethnology ; DNA, Mitochondrial ; genetics ; Gene Deletion ; Humans ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Sequence Analysis, DNA