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1.
Zhongguo Zhong Yao Za Zhi ; 49(7): 1888-1895, 2024 Apr.
Article Zh | MEDLINE | ID: mdl-38812201

This study observed the effects of Notoginseng Radix et Rhizoma on the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin complex 1(mTORC1) signaling pathway and mitochondrial energy metabolism in the rat model of adriamycin-induced renal fibrosis with blood stasis syndrome to explore the mechanism of Notoginseng Radix et Rhizoma in protecting the kidney. Thirty male rats with adriamycin-induced renal fibrosis were randomized into model, low-, medium-, and high-dose Notoginseng Radix et Rhizoma, and positive control groups(n=6). Six clean SD male rats were selected into the normal group. The normal group and model group were administrated with normal saline, and other groups with corresponding drugs. After 8 weeks of treatment, the renal function, renal pathology, adenosine triphosphate(ATP) levels, Na~+-K~+-ATPase and Ca~(2+)-Mg~(2+)-ATPase activities, and the protein levels of ATP5B, mTORC1, 70 kDa ribosomal protein S6 kinase(P70S6K), P85, Akt, p-Akt, and SH2-containing inositol phosphatase(SHIP2) in the renal tissue were determined. Compared with the normal group, the model group showed elevated levels of blood urea nitrogen(BUN) and serum creatinine(SCr)(P<0.01). Compared with the model group, Notoginseng Radix et Rhizoma and the positive control lowered the levels of BUN and SCr, which were significant in the medium-and high-dose Noto-ginseng Radix et Rhizoma groups and the positive control group(P<0.05). Compared with the model group, Notoginseng Radix et Rhizoma and the positive control alleviated the pathological changes in the renal tissue, such as vacuolar and fibroid changes, glomerulus atrophy, cystic expansion of renal tubules, and massive infiltration of inflammatory cells. Compared with the normal group, the model group showed decreased mitochondrial ATP content and Na~+-K~+-ATPase and Ca~(2+)-Mg~(2+)-ATPase activities in the renal tissue(P<0.05), and medium-and high-dose Notoginseng Radix et Rhizoma and positive control mitigated such decreases(P<0.05). Compared with the model group, medium-and high-dose Notoginseng Radix et Rhizoma and the positive control up-regulated the protein levels of ATP5B and SHIP2 and down-regulated the protein levels of mTORC1, P70S6K, P85, Akt, and p-Akt(P<0.05 or P<0.01 or P<0.001). Notoginseng Radix et Rhizoma may exert an anti-fibrosis effect by inhibiting the activation of the PI3K/Akt/mTORC1 pathway to restore mitochondrial energy metabolism, thus protecting the kidney.


Drugs, Chinese Herbal , Energy Metabolism , Mechanistic Target of Rapamycin Complex 1 , Mitochondria , Panax notoginseng , Proto-Oncogene Proteins c-akt , Rats, Sprague-Dawley , Animals , Male , Rats , Proto-Oncogene Proteins c-akt/metabolism , Proto-Oncogene Proteins c-akt/genetics , Mechanistic Target of Rapamycin Complex 1/metabolism , Panax notoginseng/chemistry , Mitochondria/drug effects , Mitochondria/metabolism , Energy Metabolism/drug effects , Drugs, Chinese Herbal/pharmacology , Drugs, Chinese Herbal/administration & dosage , Phosphatidylinositol 3-Kinases/metabolism , Phosphatidylinositol 3-Kinases/genetics , Rhizome/chemistry , Humans , Signal Transduction/drug effects , Kidney/drug effects , Kidney/metabolism , Renal Insufficiency/drug therapy , Renal Insufficiency/metabolism
2.
PLoS One ; 19(4): e0302235, 2024.
Article En | MEDLINE | ID: mdl-38635544

INTRODUCTION: Physical exercise (PE) is essential for alleviating the symptoms of sarcopenia. Low motivation is a major barrier to PE. Mindfulness-based intervention (MBI) has the potential to improve motivation. However, few studies have used a mindfulness-based PE (MBPE) intervention among older people with sarcopenia. OBJECTIVES: To assess the feasibility, acceptability and preliminary effects of the MBPE program among community-dwelling older people with sarcopenia. METHODS AND ANALYSIS: A two-arm pilot randomised controlled trial will be conducted to assess the feasibility, acceptability and preliminary effects of an MBPE program among community-dwelling older people with sarcopenia. A total of 60 participants will be randomised into the intervention group, receiving the MBPE intervention twice a week over 12 weeks, or the control group, receiving health education with the same duration, number of sessions and frequency as the intervention group. Each session of the MBPE program will last about 60 min, including 5-10- min introduction, 20-min MBI, 30-min PE and 5-10-min sharing and discussion. The primary outcomes will be the feasibility (i.e., the time spent recruiting participants, the eligibility rate and the recruitment rate) and acceptability (i.e., the attendance rate, completion rate and attrition rate) of the MBPE program. The secondary outcomes will be the preliminary effects of the MBPE program on symptoms of sarcopenia, motivation for PE, psychological well-being, mindfulness level, physical activity level and quality of life. Individual interviews will be conducted to identify the strengths, limitations and therapeutic components of the intervention. The quantitative data will be analysed by generalised estimating equations. The qualitative data will be analysed by Braun and Clarke's thematic approach. CONCLUSION: The findings of this study will be able to provide evidence for the health professionals in adopting MBPE as a supportive intervention for the older adults with sarcopenia and the groundworks for the researchers in developing non-pharmacological intervention for older adults. The positive effects could facilitate healthy ageing and relief the burden of the medical system, especially in the countries facing the ageing population. TRIAL REGISTRATION NUMBER: NCT05982067; ClinicalTrials.gov.


Mindfulness , Sarcopenia , Humans , Aged , Sarcopenia/therapy , Quality of Life , Independent Living , Feasibility Studies , Pilot Projects , Exercise , Randomized Controlled Trials as Topic
3.
Insects ; 14(2)2023 Feb 17.
Article En | MEDLINE | ID: mdl-36835769

Female weevils of the family Attelabidae (Coleoptera: Curculionoidea) possess a unique behaviour of partially cutting the branches connecting egg-bearing organs of their host plants during oviposition. However, the consequence of such behaviour remains unclear. Using Rhynchites foveipennis and its host pear (Pyrus pyrifolia), the present study tested the hypothesis that the oviposition behaviour could disarm the host plants' defence. We compared the survival rates, growth rates, and performance of eggs and larvae under two conditions: (1) the fruit stems were naturally damaged by the females before and after oviposition, and (2) the fruit stems were artificially protected from the females. When fruit stems were protected from female damage, the survival rates of eggs and larvae were only 21.3-32.6%, respectively; and the larval weight was 3.2-4.1 mg 30 days after laying eggs. When the fruit stems were damaged, the survival rates of eggs and larvae reached 86.1-94.0%, respectively; and the larval weight reached 73.0-74.9 mg 30 days after laying eggs. The contents of tannin and flavonoids in the pears did not change significantly along with the oviposition and larval feeding, but weevil eggs were crushed and killed by the callus in the pears. Once the stunted larvae in branch-growing pears were moved into the picked-off ones, the growth and development recovered. The findings indicate that the oviposition behaviour can significantly increase the survival of the offspring. Our study suggested that the oviposition behaviour of attelabid weevils is a strategy to overcome plant defence.

4.
Chin J Integr Med ; 29(5): 434-440, 2023 May.
Article En | MEDLINE | ID: mdl-36474083

OBJECTIVE: To investigate the effect and potential mechanism of dihydromyricetin (Dmy) on H9C2 cell proliferation, apoptosis, and autophagy. METHODS: H9C2 cells were randomly divided into 7 groups, namely control, model, EV (empty pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro vector), IV (circHIPK3 interference), Dmy (50 µ mol/L), Dmy+IV, and Dmy+EV groups. Cell proliferation and apoptosis were detected by cell counting kit-8 assay and flow cytometry, respectivley. Western blot was used to evaluate the levels of light chain 3 II/I (LC3II/I), phospho-phosphoinositide 3-kinase (p-PI3K), protein kinase B (p-AKT), and phospho-mammalian target of rapamycin (p-mTOR). The level of circHIPK3 was determined using reverse transcriptase polymerase chain reaction. Electron microscopy was used to observe autophagosomes in H9C2 cells. RESULTS: Compared to H9C2 cells, the expression of circHIPK in H9C2 hypoxia model cells increased significantly (P<0.05). Compared to the control group, the cell apoptosis and autophagosomes increased, cell proliferation rate decreased significantly, and the expression of LC3 II/I significantly increased (all P<0.05). Compared to the model group, the rate of apoptosis and autophagosomes in IV, Dmy, and Dmy+IV group decreased, the cell proliferation rate increased, and the expression of LC3 II/I decreased significantly (all P<0.05). Compared to the control group, the expressions of p-PI3K, p-AKT, and p-mTOR in the model group significantly reduced (P<0.05), whereas after treatment with Dmy and sh-circHIPK3, the above situation was reversed (P<0.05). CONCLUSION: Dmy plays a protective role in H9C2 cells by inhibiting circHIPK expression and cell apoptosis and autophagy, and the mechanism may be related to PI3K/AKT/mTOR pathway.


Proto-Oncogene Proteins c-akt , Signal Transduction , Proto-Oncogene Proteins c-akt/metabolism , Phosphatidylinositol 3-Kinases/metabolism , TOR Serine-Threonine Kinases/metabolism , Apoptosis , Autophagy
5.
Purinergic Signal ; 19(1): 43-53, 2023 03.
Article En | MEDLINE | ID: mdl-35389158

This study explored the role of P2X7 receptors in spinal cord astrocytes in the electroacupuncture-induced inhibition of visceral hypersensitivity (VH) in rats with irritable bowel syndrome (IBS). Visceral hypersensitivity of IBS was intracolonically induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS). Visceromotor responses to colorectal distension (CRD-20,40,60,80 mmHg) and abdominal withdrawal reflex scoring (AWRs) were recorded after electroacupuncture at bilateral Zusanli (ST36) and Sanyinjiao (SP6) acupoints to evaluate the analgesic effect of electroacupuncture on visceral pain in rats with IBS. Fluorocitric acid (FCA), an astrocyte activity inhibitor, was injected intrathecally before electroacupuncture intervention and AWRs were recorded. Western blot and real-time qPCR were used to detect the expression of NMDA and P2X7 receptor to observe the regulation effect of electroacupuncture on NMDA receptor in the spinal cord of rats with visceral hypersensitivity. Intrathecal injection of P2X7 agonist or antagonist was administered before electroacupuncture treatment. To observe the effect of P2X7 receptor in spinal astrocytes on the inhibition of visceral hyperalgesia by electroacupuncture, the changes of AWR score, NMDA receptor in the spinal cord, and GFAP expression in astrocytes were detected. Inflammation of the colon had basically subsided at day 21 post-TNBS; persistent visceral hypersensitivity could be suppressed by electroacupuncture. This analgesic effect could be inhibited by FCA. The analgesic effect, downregulation of NMDA receptor NR1 subunit, and P2X7 protein of electroacupuncture were all reversed by FCA. P2X7 receptor antagonist A740003 can cooperate with EA to carry out analgesic effect in rats with visceral pain and downregulate the expression of NR1, NR2B, and GFAP in spinal dorsal horn. However, the P2X7 receptor agonist BzATP could partially reverse the analgesic effect of EA, inhibiting the downregulatory effect of EA on the expression of NR1, NR2B, and GFAP. These results indicate that EA may downregulate the expression of the NMDA receptor by inhibiting the P2X7 receptor in the spinal cord, thereby inhibiting spinal cord sensitization in IBS rats with visceral pain, in which astrocytes are an important medium.


Electroacupuncture , Hypersensitivity , Irritable Bowel Syndrome , Visceral Pain , Rats , Animals , Irritable Bowel Syndrome/metabolism , Irritable Bowel Syndrome/therapy , Rats, Sprague-Dawley , Astrocytes/metabolism , Visceral Pain/metabolism , Electroacupuncture/methods , Receptors, N-Methyl-D-Aspartate/metabolism , Receptors, Purinergic P2X7/metabolism , Spinal Cord/metabolism , Spinal Cord Dorsal Horn/metabolism , Hypersensitivity/metabolism , Analgesics
6.
Yi Chuan ; 44(8): 708-719, 2022 Aug 20.
Article En | MEDLINE | ID: mdl-36384669

The fast-rising CRISPR-derived gene editing technologies has been widely used in the fields of life science and biomedicine, as well as plant and animal breeding. However, the efficiency of homology-directed repair (HDR), an important strategy for gene knock-in and base editing, remains to be improved. In this study, we came up with the term Donor Adapting System (DAS) to summarize those CRISPR/Cas9 systems modified with adaptor for driving aptamer-fused donor DNA. A set of CRISPR/Cas9-Gal4BD DAS was designed in our study. In this system, Gal4 DNA binding domain (Gal4BD) is used as adaptor to fuse with Cas9 protein, and Gal4 binding sequence (Gal4BS) is used as aptamer to bind to the double-stranded DNA (dsDNA) donor, in order to improve the HDR efficiency. Preliminary results from the HEK293T-HDR.GFP reporter cell line show that the HDR editing efficiency could be improved up to 2-4 times when donor homologous arms under certain length (100-60 bp). Further optimization results showed that the choice of fusion port and fusion linker would affect the expression and activity of Cas9, while the Cas9-Gal4BD fusion with a GGS5 linker was the prior choice. In addition, the HDR efficiency was likely dependent on the aptamer-dsDNA donor design, and single Gal4BD binding sequence (BS) addition to the 5'-end of intent dsDNA template was suggested. Finally, we achieved enhanced HDR editing on the endogenous AAVS1 and EMX1 sites by using the CRISPR/Gal4BD-Cas9 DAS, which we believe can be applied to facilitate animal molecular design breeding in the future.


CRISPR-Cas Systems , Recombinational DNA Repair , Animals , Humans , DNA , HEK293 Cells
7.
Anal Methods ; 14(6): 672-677, 2022 02 11.
Article En | MEDLINE | ID: mdl-35088063

The sensitive determination of the critical micelle concentration (CMC) of surfactants is very important for their practical application. Due to their good sensitivity and simple operation, pyrene and its derivatives have been widely used as fluorescent probes to detect the CMC. However, their virulent and poor water-soluble nature has limited their wide employment. In the present work, environmentally friendly amphiphilic carbon dots (Cdots) with dual-color emission and absolute quantum yield (PLQY) values higher than 50% have been fabricated through a solvothermal process, which could successfully serve as self-calibrative, ratiometric fluorescent probes to estimate the CMC of both non-ionic and ionic surfactants. This work not only provides a new strategy to design green ratiometric fluorescent probes for the CMC measurement of surfactants but also expands the application of Cdots in the colloidal field.


Carbon , Quantum Dots , Fluorescent Dyes , Micelles , Surface-Active Agents
8.
Chin Med J (Engl) ; 134(13): 1576-1583, 2021 Jun 16.
Article En | MEDLINE | ID: mdl-34133352

BACKGROUND: Various prediction tools have been developed to predict biochemical recurrence (BCR) after radical prostatectomy (RP); however, few of the previous prediction tools used serum prostate-specific antigen (PSA) nadir after RP and maximum tumor diameter (MTD) at the same time. In this study, a nomogram incorporating MTD and PSA nadir was developed to predict BCR-free survival (BCRFS). METHODS: A total of 337 patients who underwent RP between January 2010 and March 2017 were retrospectively enrolled in this study. The maximum diameter of the index lesion was measured on magnetic resonance imaging (MRI). Cox regression analysis was performed to evaluate independent predictors of BCR. A nomogram was subsequently developed for the prediction of BCRFS at 3 and 5 years after RP. Time-dependent receiver operating characteristic (ROC) curve and decision curve analyses were performed to identify the advantage of the new nomogram in comparison with the cancer of the prostate risk assessment post-surgical (CAPRA-S) score. RESULTS: A novel nomogram was developed to predict BCR by including PSA nadir, MTD, Gleason score, surgical margin (SM), and seminal vesicle invasion (SVI), considering these variables were significantly associated with BCR in both univariate and multivariate analyses (P < 0.05). In addition, a basic model including Gleason score, SM, and SVI was developed and used as a control to assess the incremental predictive power of the new model. The concordance index of our model was slightly higher than CAPRA-S model (0.76 vs. 0.70, P = 0.02) and it was significantly higher than that of the basic model (0.76 vs. 0.66, P = 0.001). Time-dependent ROC curve and decision curve analyses also demonstrated the advantages of the new nomogram. CONCLUSIONS: PSA nadir after RP and MTD based on MRI before surgery are independent predictors of BCR. By incorporating PSA nadir and MTD into the conventional predictive model, our newly developed nomogram significantly improved the accuracy in predicting BCRFS after RP.


Nomograms , Prostatic Neoplasms , Humans , Male , Neoplasm Grading , Neoplasm Recurrence, Local/diagnosis , Neoplasm Recurrence, Local/surgery , Prognosis , Prostate-Specific Antigen , Prostatectomy , Prostatic Neoplasms/surgery , Retrospective Studies , Seminal Vesicles
9.
World J Gastroenterol ; 26(39): 5997-6014, 2020 Oct 21.
Article En | MEDLINE | ID: mdl-33132650

BACKGROUND: Autophagy is an evolutionarily conserved biological process in eukaryotic cells that involves lysosomal-mediated degradation and recycling of related cellular components. Recent studies have shown that autophagy plays an important role in the pathogenesis of Crohn's disease (CD). Herbal cake-partitioned moxibustion (HM) has been historically practiced to treat CD. However, the mechanism by which HM regulates colonic autophagy in CD remains unclear. AIM: To observe whether HM can alleviate CD by regulating colonic autophagy and to elucidate the underlying mechanism. METHODS: Rats were randomly divided into a normal control (NC) group, a CD group, an HM group, an insulin + CD (I + CD) group, an insulin + HM (I + HM) group, a rapamycin + CD (RA + CD) group, and a rapamycin + HM (RA + HM) group. 2,4,6-trinitrobenzenesulfonic acid was administered to establish a CD model. The morphology of the colonic mucosa was observed by hematoxylin-eosin staining, and the formation of autophagosomes was observed by electron microscopy. The expression of autophagy marker microtubule-associated protein 1 light chain 3 beta (LC3B) was observed by immunofluorescence staining. Insulin and rapamycin were used to inhibit and activate colonic autophagy, respectively. The mRNA expression levels of phosphatidylinositol 3-kinase class I (PI3KC1), Akt1, LC3B, sequestosome 1 (p62), and mammalian target of rapamycin (mTOR) were evaluated by RT-qPCR. The protein expression levels of interleukin 18 (IL-18), tumor necrosis factor-α (TNF-α), nuclear factor κB/p65 (NF-κB p65), LC3B, p62, coiled-coil myosin-like BCL2-interacting protein (Beclin-1), p-mTOR, PI3KC1, class III phosphatidylinositol 3-kinase (PI3KC3/Vps34), and p-Akt were evaluated by Western blot analysis. RESULTS: Compared with the NC group, the CD group showed severe damage to colon tissues and higher expression levels of IL-18 and NF-κB p65 in colon tissues (P < 0.01 for both). Compared with the CD group, the HM group showed significantly lower levels of these proteins (P IL-18 < 0.01 and P p65 < 0.05). There were no significant differences in the expression of TNF-α protein in colon tissue among the rat groups. Typical autophagic vesicles were found in both the CD and HM groups. The expression of the autophagy proteins LC3B and Beclin-1 was upregulated (P < 0.01 for both) in the colon tissues of rats in the CD group compared with the NC group, while the protein expression of p62 and p-mTOR was downregulated (P < 0.01 for both). However, these expression trends were significantly reversed in the HM group compared with the CD group (P LC3B < 0.01, P Beclin-1 < 0.05, P p62 < 0.05, and P m-TOR < 0.05). Compared with those in the RA + CD group, the mRNA expression levels of PI3KC1, Akt1, mTOR, and p62 in the RA + HM group were significantly higher (P PI3KC1 < 0.01 and P Akt1, mTOR, and p62 < 0.05), while those of LC3B were significantly lower (P < 0.05). Compared with the RA + CD group, the RA + HM group exhibited significantly higher PI3KC1, p-Akt1, and p-mTOR protein levels (P PI3KC1 < 0.01, P p-Akt1 < 0.05, and P p-mTOR < 0.01), a higher p62 protein level (P = 0.057), and significantly lower LC3B and Vps34 protein levels (P < 0.01 for both) in colon tissue. CONCLUSION: HM can activate PI3KC1/Akt1/mTOR signaling while inhibiting the PI3KC3 (Vps34)-Beclin-1 protein complex in the colon tissues of CD rats, thereby inhibiting overactivated autophagy and thus exerting a therapeutic effect.


Biological Phenomena , Crohn Disease , Moxibustion , Animals , Autophagy , Colon , Crohn Disease/therapy , Phosphatidylinositol 3-Kinases , Rats
10.
J Poult Sci ; 56(1): 27-31, 2019 Jan 25.
Article En | MEDLINE | ID: mdl-32055193

Improving feed efficiency is important for decreasing feed cost in poultry production, because feed account for approximately 70% of the total production costs. The selection of feed efficiency may affect other important economic traits. Therefore, the objectives of this present study was to evaluate the relationships of the residual feed intake (RFI) with live body weight, carcass weight, carcass composition, and size of small intestines in a population of F2 Pekin ducks. Nine-hundred and eighty F2 ducks were derived from a cross between 40 Pekin ducks and 10 Mallard ducks. The results showed no significant correlation of RFI with live body weight and eviscerated carcass weight. RFI had negative effects on breast meat weight and gizzard weight. A positive correlation of RFI with abdominal fat weight, skin weight, and jejunum length was detected. Our results indicated that the selection of RFI could improve the feed efficiency of ducks without affecting their carcass compositions.

12.
Nat Commun ; 9(1): 4488, 2018 10 26.
Article En | MEDLINE | ID: mdl-30367062

Today East Asia harbors many "relict" plant species whose ranges were much larger during the Paleogene-Neogene and earlier. The ecological and climatic conditions suitable for these relict species have not been identified. Here, we map the abundance and distribution patterns of relict species, showing high abundance in the humid subtropical/warm-temperate forest regions. We further use Ecological Niche Modeling to show that these patterns align with maps of climate refugia, and we predict species' chances of persistence given the future climatic changes expected for East Asia. By 2070, potentially suitable areas with high richness of relict species will decrease, although the areas as a whole will probably expand. We identify areas in southwestern China and northern Vietnam as long-term climatically stable refugia likely to preserve ancient lineages, highlighting areas that could be prioritized for conservation of such species.

13.
Sci Rep ; 7: 43822, 2017 03 08.
Article En | MEDLINE | ID: mdl-28272437

This study, using species distribution modeling (involving a new approach that allows for uncertainty), predicts the distribution of climatically suitable areas prevailing during the mid-Holocene, the Last Glacial Maximum (LGM), and at present, and estimates the potential formation of new habitats in 2070 of the endangered and rare Tertiary relict tree Davidia involucrata Baill. The results regarding the mid-Holocene and the LGM demonstrate that south-central and southwestern China have been long-term stable refugia, and that the current distribution is limited to the prehistoric refugia. Given future distribution under six possible climate scenarios, only some parts of the current range of D. involucrata in the mid-high mountains of south-central and southwestern China would be maintained, while some shift west into higher mountains would occur. Our results show that the predicted suitable area offering high probability (0.5‒1) accounts for an average of only 29.2% among the models predicted for the future (2070), making D. involucrata highly vulnerable. We assess and propose priority protected areas in light of climate change. The information provided will also be relevant in planning conservation of other paleoendemic species having ecological traits and distribution ranges comparable to those of D. involucrata.


Climate Change , Ecosystem , Endangered Species , Nyssaceae/growth & development , Refugium , Trees/growth & development , China , Conservation of Natural Resources/methods , Conservation of Natural Resources/trends , Geography , Models, Theoretical , Population Dynamics
14.
Yi Chuan ; 38(1): 28-39, 2016 01.
Article Zh | MEDLINE | ID: mdl-26787521

Targeted genome editing technology plays an important role in studies of gene function, gene therapy and transgenic breeding. Moreover, the efficiency of targeted genome editing is increased dramatically with the application of recently developed artificial nucleases such as ZFNs, TALENs and CRISPR/Cas9. However, obtaining positive cells with targeted genome modification is restricted to some extent by nucleases expression plasmid transfection efficiency, nucleases expression and activity, and repair efficiency after genome editing. Thus, the enrichment and screening of positive cells with targeted genome modification remains a problem that need to be solved. Surrogate reporter systems could be used to reflect the efficiency of nucleases indirectly and enrich genetically modified positive cells effectively, which may increase the efficiency of the enrichment and screening of positive cells with targeted genome modification. In this review, we mainly summarized principles and applications of reporter systems based on NHEJ and SSA repair mechanisms, which may provide references for related studies in future.


Gene Targeting/trends , Genes, Reporter , Genetic Engineering/trends , Genome , Animals , Gene Targeting/methods , Genetic Engineering/methods , Humans
15.
Yi Chuan ; 37(10): 1053-60, 2015 10.
Article En | MEDLINE | ID: mdl-26496758

Reporter vector system has become an important method for measuring activity of specific nucleases because of its fast construction, simple modification, easy operation, economic effectiveness as well as its role in enriching positive cells with genomic modification through mediating screen of specific nucleases positive cells. After introducing double strand breaks (DSBs), a reporter system based on non-homologous end joining (NHEJ)-mediated repair can only repair maximally two thirds of reporter genes after optimization, while single strand annealing (SSA)-mediated repair can repair all reporter genes theoretically which has higher sensitivity and facilitates the detection of specific nuclease with low activity and provides an effective way to detect specific nuclease activity in genome modification studies. In this study, we designed and constructed three sets of dual-fluorescence reporter systems based on SSA repair mechanism and applied the mRFP-eGFP system in measuring the effective activity of three pairs of ZFNs, which was 8.9%, 9.3% and 5.0%, respectively. Our study provides an effective way to detect the activity of nucleases.


Endonucleases/metabolism , Flow Cytometry/methods , Green Fluorescent Proteins/metabolism , Luminescent Proteins/metabolism , Microscopy, Fluorescence, Multiphoton/methods , DNA Repair , DNA, Single-Stranded/genetics , Endonucleases/genetics , Enzyme Assays/methods , Green Fluorescent Proteins/genetics , HEK293 Cells , Humans , Luminescent Proteins/genetics , Reproducibility of Results , Zinc Fingers/genetics , Red Fluorescent Protein
16.
Biomed Environ Sci ; 28(6): 464-7, 2015 Jun.
Article En | MEDLINE | ID: mdl-26177909

In order to evaluate the performance of a molecular Hain line probe assay (Hain LPA) for rapid detection of rifampicin and isoniazid resistance of Mycobacterium tuberculosis in China, 1612 smear positive patients were consecutively enrolled in this study. Smear positive sputum specimens were collected for Hain LPA and conventional drug susceptibility testing (DST). The sensitivity and specificity of Hain LPA were analyzed by using conventional DST as golden reference. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for rifampicin resistance detection were 88.33%, 97.66%, 81.54%, and 98.62%, respectively. The sensitivity, specificity, PPV and NPV for isoniazid resistance detection were 80.25%, 98.07%, 87.25%, and 96.78%, respectively. These findings suggested that Hain LPA can be an effective method worthy of broader use in China.


Genotyping Techniques/methods , Mycobacterium tuberculosis/drug effects , Mycobacterium tuberculosis/genetics , Tuberculosis, Multidrug-Resistant/microbiology , China , Humans , Isoniazid/pharmacology , Mycobacterium tuberculosis/isolation & purification , Rifampin/pharmacology , Tuberculosis, Multidrug-Resistant/diagnosis
17.
Stem Cells Int ; 2015: 941625, 2015.
Article En | MEDLINE | ID: mdl-25861281

Effective repair of peripheral nerve defects is difficult because of the slow growth of new axonal growth. We propose that "neural-like cells" may be useful for the protection of peripheral nerve destructions. Such cells should prolong the time for the disintegration of spinal nerves, reduce lesions, and improve recovery. But the mechanism of neural-like cells in the peripheral nerve is still unclear. In this study, bone marrow-derived neural-like cells were used as seed cells. The cells were injected into the distal end of severed rabbit peripheral nerves that were no longer integrated with the central nervous system. Electromyography (EMG), immunohistochemistry, and transmission electron microscopy (TEM) were employed to analyze the development of the cells in the peripheral nerve environment. The CMAP amplitude appeared during the 5th week following surgery, at which time morphological characteristics of myelinated nerve fiber formation were observed. Bone marrow-derived neural-like cells could protect the disintegration and destruction of the injured peripheral nerve.

18.
J Insect Sci ; 152015.
Article En | MEDLINE | ID: mdl-25791200

Mechoris ursulus (Roelofs) (Coleoptera: Attelabidae) is a pest weevil of Fagaceae oak trees in eastern Asia. The female has a distinct branch-cutting behavior in conjunction with oviposition in the acorns of its host plant. This study analyzed the factors influencing oviposition preference by carrying out continuous field surveys over the course of 2009-2010 and through laboratory rearing. The field survey showed that for both of the hosts, Cyclobalanopsis glaucoides Schottky (Fagales: Fagaceae) and Quercus franchetii Skan, M. ursulus preferred branches with fewer acorns and larger acorns on the same branch for oviposition. Laboratory rearing experiments showed offspring performance (i.e., survival rate and fresh weight of larvae) was significantly and positively correlated with acorn size. Preference for larger acorns could maximize the fitness of offspring by providing sufficient food source and space.


Coleoptera/physiology , Fagaceae/growth & development , Food Chain , Oviposition , Animals , China , Coleoptera/growth & development , Female , Larva/growth & development , Larva/physiology , Quercus/growth & development
19.
Biomed Environ Sci ; 27(12): 982-6, 2014 Dec.
Article En | MEDLINE | ID: mdl-25484017

Sputum transportation from county-level to prefecture-level is an ideal strategy to cover the shortage of the laboratory capability in the resource-poor setting. Here, we firstly evaluated the feasibility of sputum transportation system in China by analyzing the culture and molecular diagnosis results from 1982 smear-positive patients with different delay in processing for culture. In this study, the total contamination rate was 2.32% and the total smear positive/culture negative (S+/C-) rate was 7.57%. We found that sputum specimens refrigerated for no more than 7 d before mycobacterial detection did not affect culture significantly. In addition, the invalid result rates among 0-3 d, 3-7 d, and 7+ d group were 3.63%, 3.14%, and 12.48%, respectively. Statistic analysis revealed that molecular diagnostic results while the invalid result rate of genechip for the specimen with more than 7 d delay was significantly higher (P<0.001). The refrigerators equipped in county laboratories, transport at low temperature and frequent transport services once a week will ensure the feasibility of sputum transportation system in China.


Mycobacterium/isolation & purification , Specimen Handling , Sputum/microbiology , China , Feasibility Studies , Humans , Transportation
20.
Gene ; 538(2): 354-60, 2014 Apr 01.
Article En | MEDLINE | ID: mdl-24463020

LIM homeobox transcription factor 4 (LHX4) is a LIM homeodomain transcription factor involved in pituitary gland and nervous system development. The aim of this study was to examine the association of the LHX4 polymorphisms with growth traits in beef cattle breed. A total of 7 single nucleotide polymorphyisms (SNPs) have been identified in the coding region and noncoding region of the bovine LHX4 by sequencing pooled DNA samples (Pool-Seq) and PCR-single strand conformation polymorphism (PCR-SSCP) methods. The linkage disequilibrium was assessed in 871 individuals representing four main cattle breeds from China. The SNPs 2-5 and 7-8 were found to be in complete linkage disequilibrium, respectively. The result of haplotype analysis of 13 SNPs showed that 31 haplotypes were found in four Chinese cattle breeds, and 20 genotypes were only found in Nanyang cattle. The statistical analyses indicated that the SNP1-5, and 6 are associated with the body weight at 18, and 6 months of age in Nanyang cattle population (P<0.05), but no significant associations between their twenty combined genotypes. Our results provide evidence that some polymorphisms in LHX4 are associated with growth traits at certain ages, and may be used as candidates for marker-assisted selection and management in cattle.


Cattle/growth & development , Cattle/genetics , LIM-Homeodomain Proteins/genetics , Age Factors , Animals , Body Weight/genetics , Breeding , China , Genetic Markers , Genetic Variation , Haplotypes , Linkage Disequilibrium , Polymorphism, Single Nucleotide , Polymorphism, Single-Stranded Conformational , Species Specificity
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