BACKGROUND: The target gene cannot be deleted at specific time by using conditional gene knockout system. Most gene knockout animals cannot survive during embryonic development and after birth, which limits the study of gene function.
OBJECTIVE: To review the recent advances about time-specific gene knockout technology.
METHODS: An online search in PubMed was performed from the initial stage of database establishment to August 2014. The key words were gene knockout, inducer and Cre/loxP system. The repeated experiment and unrelated articles were excluded during primary search. A summary was made by analyzing time-specific gene knockout technology.
RESULTS AND CONCLUTION: Most important gene knockout animals could not survive after birth, or die at early embryonic stage due to serious physical defects induced by an important gene deletion. Therefore, the progress in the functionality of a gene at a particular time, the genetic characteristics of an individual organism in a particular developmental period, and the treatment of certain acquired disease at a given time have been limited because of the limitations of research methods. However, the occurrence of conditional gene knockout animals, especial y time-specific knockout animals, provides a research model for the study of these areas. This study reviewed the recent advances, development and application about time-specific gene knockout animals, and discussed the significance and broad prospects of time-specific gene knockout technology of specific genes in the field of scientific research.

In recent years, the eradication rate of standard triple therapy for Helicobacter pylori (Hp) infection has been reduced to less than 80% because of many factors, such as strain variation, drug resistance and cross infection of different strains.As more and more cases with failure of initial eradication therapy, the focus of clinical practice is to explore new alternative rescue therapies.Aims: To assess the efficacy and safety of Saccharomyces boulardii combined with moxifloxacin-based triple regimen for rescue therapy of Hp infection.Methods: A total of 400 patients with chronic gastritis who had failed initial bismuth quadruple therapy were enrolled and randomly assigned into two groups: 200 cases of control group and 200 cases of test group.Patients in control group received a rescue triple therapy (esomeprazole, amoxicillin and moxifloxacin) for 14 days and those in test group with the further addition of Saccharomyces boulardii sachets.Hp eradication was assessed by 13C/14C-urea breath test four weeks after treatment, and the adverse events during treatment course were observed.Results: The Hp eradication rates in control group and test group were 80.1% (121/151) and 90.4% (142/157) by per-protocol (PP) analysis, respectively, and 60.5% (121/200) and 71.0% (142/200) by intention-to-treat (ITT) analysis, respectively.The eradication rate in test group was significantly higher than that in control group by both PP and ITT analyses (P<0.05).The incidence rate of adverse events was significantly lower in test group than in control group (9.6% vs.17.9%, P<0.05).Conclusions: Saccharomyces boulardii added to moxifloxacin-based rescue triple therapy could increase the eradication rate of Hp infection with less adverse events.

ObjectiveTo observe the clinical efficacy of relaxing needling plus joint needling from the Internal Medicine in treating post-stroke spastic hemiplegia.MethodTotally 121 eligible subjects were randomized intoan ordinary acupuncture group (61 cases) and a relaxing-joint needling group (60 cases). The two groups were treated for 2 weeks, 5 sessions each week. The spasm of the lower limb was evaluated by using Fugl-Meyer Assessment Scale (FMA), modified AshworthScale (MAS), Tardieu Scale, and Activities of Daily Living (ADL) before and after treatment. Besides, 4 patients from each group were randomly selected for the three-dimensional gait analysis, and the dorsi-extension, range of motion (ROM) of ankle, strephenopodia, and heel-raising height were observed.Result and ConclusionRelaxing-joint needling and ordinary acupuncture both can produce significant efficacies in treating post-stroke spastic hemiplegia. The relaxing-joint needling produced more significant efficacies in improving the body balance, ROM, muscular tension, and lower limb spasticity compared to ordinary acupuncture. It’s plausible that relaxing-joint needling improves the strephenopodia and hemiplagia gait through renovating body balance, ROM, muscular tension, and lower limb spasticity. Since the three-dimensional gait analysis can analyze the dorsi-extension, strephenopodia, and heel-raising height, thus it can be used to evaluate the change of the spastic gait of post-stroke hemiplegia in amore objective, elaborate, and correct way.

Objective To investigate the effects of sensorimotor therapy on patients with knee osteoarthritis.Methods Ninety-two patients with knee osteoarthritis were divided into a treatment group and a control group by using random digits table. Both groups received conventional rehabilitation therapy, including knee isometric and isotonic contraction training, ultrashort wave diathermy as well as aerobic exercise. The treatment group was also administered with sensorimotor training, in addition to the conventional training,including the muscle strengthening and control training of quadriceps and hamstring muscle using sling exercise therapy system and on the Thera-band mattress. The visual analogue scale (VAS) for pain, manual muscle testing and knee range of motion as well as the Japanese Orthopedic Association knee function evaluation form were used to evaluate the patients. Results There was no statistic difference between the 2 groups before treatment, with regard to the evaluation parameters used in this study. After 2 months of treatment, both groups improved significantly (P ＜0.05),with the treatment group improved to a significantly greater extent (P ＜0.05). Conclusions Sensorimotor training in addition to the conventional rehabilitation treatment can further relieve the pain of patients with knee osteoarthritis joints arthritis and improve the function of patients with knee osteoarthritis.

Objective To investigate the technical feasibility and oncological safety of EPMR for early esophageal cancer and precancerous lesions. Methods Clinicopathological data, procedure quality and complications of 70 patients with early esophageal cancer or precancerous lesions were retrospectively ana?lysed. The technical safety and feasibility were assessed, and the complications were investigated through postoperative surveillance. The results were compared with published data from two high?quality meta?analysis studies on ESD. Results In a total of 70 patients who underwent EPMR, 35?7%( 25/70) were diagnosed as having early esophageal cancer and 64?3%( 45/70) atypical hyperplasia. And tumor size greater than 2 cm was 78?6%(55/70). The mean EPMR time was(28?31±9?49) minutes. The R0 resection rate of EPMR was 88?6%(62/70) and no perforation occurred. The bleeding rate was 21?4%(15/70). The esoph?ageal stricture rate was 1?4%( 1/70) . The recurrence rate was 2?9%( 2/70) . Compared with published ESD data from the two studies, EPMR showed shorter procedure time, lower curative rate and lower rates of stric?ture and perforation, but slightly higher rates of bleeding and recurrence. Conclusion Both EPMR and ESD are feasible and effective for early esophageal cancer and precancerous lesions. The curative results of EPMR are similar to ESD, with shorter procedure time, but higher bleeding and recurrence rates.

Objective To investigate whether Th17/Treg imbalance existed,and whether VEGF165 attenuated the imbalance in allogeneic skeletal myoblasts transplantation(allo-SMT) for acute myocardial infarction (AMI).Methods There were 60 C57BL/6 male mice,3 to 4 weeks old,weighting 0.016-0.020 kg.On the day 1,2,4,and 7 after allo-SMT,the percentages of Th17 and Treg cells,and the ratios of Th17/Treg cells were analyzed by FCM in AMI group,AMI-S group(alloSMT) and AMI-Ⅴ group(with VEGF165 treatment).Subsequently,related proinflammatory and regulatory cytokines and key transcription factors ROR-γt mRNA and Foxp3 mRNA expression were examined by Bio-plex and RT-PCR respectively.Results On the day 1,2,4,and 7 after allo-SMT,the percentage of Treg,related cytokines concentrations and transcript factor Foxp3 mRNA in AMI-S group were lower than those in AMI group,while the AMI-Ⅴ group were higher than those in AMI groups.However,the percentage of Th17 cells,related cytokines concentrations,and ROR-γt mRNA in AMI-S group were higher than those in AMI group; but the AMI-Ⅴ group were lower than those in AMI group.Apparently,compared with AMI group,the ratios of Th17/Treg significantly ascended in AMI-S group and decended in AMI-Ⅴ group.Conclusion Th17/Treg imbalance participated in the formation and development of inflammation and immune response after allo-SMT.However,transfected VEGF165 could relieve the severity of Th17/Treg imbalance.

BACKGROUND:Differentiation of growth factor-induced cells into fibrocartilage is the hot topic in the research of meniscus tissue engineering. The in vitro construction and in vivo reconstruction of menisci are closely related with the effect of growth factors. OBJECTIVE:To review the current development of growth factors in meniscus tissue engineering, and to investigate the mechanism. METHODS:The VIP database (http://lib.cqvip.com), CNKI database (www.cnki.net) and PubMed database (http://www.ncbi.nlm.nih.gov/pubmed) were retrieved for the related articles from January 2008 to March 2013 with the key words of“meniscus tissue engineering, cartilage, growth factors”. A total of 53 articles on the research of growth factors in meniscus tissue engineering were included. RESULTS AND CONCLUSION:There were various growth factors in cartilage tissue engineering research, and the new growth factors have been found in the constant. The research on the regulation effect of growth factors on cartilage was changed from the research of single growth factor model to the research of the interactions between multiple growth factors;the molecular mechanisms of growth factors in regulating the cartilage have also been extensively studied. Growth factors have good application prospects in tissue engineering, but there are stil many problems to be solved, such as during the healing of menisci, the expressions and the effect of different growth factors are different in different time points. So how to play the interaction between growth factors timely and appropriately thus stimulating the in vivo microenvironment, investigating the molecular mechanisms of growth factors in regulating the cartilage, as wel as finding the new growth factors are the research focus in meniscus tissue engineering.

Objective To investigate the effects of meliorated renal failure decoction (MRFD) on renal function and inflammatory cytokines in patients with CKD 3-4 stages. Methods Sixty-two patients with CKD 3-4 stages were divided into treatment group and control group through random number table method. Both groups were given conventional treatment of western medicine, and the treatment group was given MRFD additionally. After 6 months of treatment, the changes of TCM symptom scores, Hb, Scr, BUN, 24 h UP, IL-6, IL-8, TNF and eGFR were observed. Meanwhile, the clinical efficacy was evaluated. Results The total clinical effective rate in treatment group was 65.63%(21/32), and 30.00%(9/30) in the control group (P<0.01);the effective rate of TCM syndromes in treatment group was 96.88%(31/32), and 53.33%(16/30) in the control group (P<0.01). The symptoms were improved better in the treatment group than the control group except for the symptoms of dry mouth and throat, and diarrhea. The level of eGFR in treatment group increased obviously (P<0.05), while the levels of Scr, 24 h UP and BUN decreased (P<0.05, P<0.01), with statistical significance between the two groups ( P<0.01). The level of eGFR increased, and the levels of Scr decreased in the control group (P<0.01). The levels of IL-6, IL-8, and TNF in both groups were reduced, while the level of Hb increased in the two groups, without statistical significance (P>0.05). Conclusion MRFD can improve TCM syndrome and renal function, delay the progress of patients with CKD at 3-4 stage by means of reducing the inflammatory cytokines and inhibiting micro-inflammatory state.

Background Different depthes or distances are prompted with many visual cues on the flat screen.Which kind of depth cues plays a major role in the accommodation of human eyes,in addition,whether this effect exists in the different distance are still below understood.Objective This study was to research the effect of the monocular space depth perception on accommodative response at different distances,and to analyze the effect of linear perspective and size-constancy perceptual depth cues on the level and amplitude of accommodative fluctation.Methods Four groups of visual targets were designed by FLASH software and placed at the front of eyes with the distance of 50 cm or 33.3 cm,representing static change from far to near and dynamic change from far to near,from near to far respectively.Twenty-six healthy volunteers aged 22-26 years were included in this study,with the spherical power of (-4.06± 1.99) D and cylindrical power of (-0.38±0.41) D under the informed consent.The accommodative response and accommodative fluctuations during monocularly watching the targets were recorded at 5 times/second by Grand Seiko WAM 5500 automatic infrared refractor.This study was approved by Ethic Committee of Wenzhou Medical University,and written informed consent was obtained from each subject before entering the study group.Results At the distance of 50 cm,the accommodative response caused by the depth perception of near was (1.52±0.46) D,the one of distance was (1.37±0.46) D,with a significant difference of 0.15 D under the combined effect of linear perspective and size-constancy (P =0.016).The accommodative response caused by the depth perception of big car was (1.43 ± 0.35) D,and the one of small car was (1.43 ± 0.36) D,without statistically significant difference under the effect of size-constancy alone (P =0.467).At the distance of 33.3 cm,the accommodative response caused by the depth perception of near was (2.40±0.53)D,and the one of distance was (2.35 ±0.51) D,without significant difference under the combined effect of linear perspective and size-constancy (P=0.379).The accommodative response caused by the depth perception of big car was (2.38±0.48)D,the one of small car was (2.39±0.52)D,without statistically significant difference under the effect of size-constancy alone (P =0.820).In addition,the total curve of accommodative microfluctuation showed apparent fluctuation periodically caused by the distance perception involving linear perspective only.Conclusions Monocular space depth perception has some impacts on the accommodative response at certain distance.Linear perspective,which is one of the depth cues of space perception,plays a major role in the accommodative response.Compared with the size-constancy,linear perspective can provide depth perception at higher level.The size-constancy has a little or contrast impact to the accommodation,because it depends on the assistance from other depth cues like linear perspective.