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Tomato ( Solanum lycopersicum ) is a globally cultivated crop with great economic value. The exocarp determines the appearance of tomato fruit and protects it from various biotic and abiotic challenges at both pre-harvest and post-harvest stages. However, no tomato exocarp-specific promoter is currently available, which hinders exocarp-based genetic engineering. Here, we identified by RNA sequencing and reverse transcription-quantitative PCR analyses that the tomato gene SlPR10 ( PATHOGENESIS RELATED 10 ) was abundantly and predominantly expressed in the exocarp. A fluorescent reporter expressed by a 2087-bp SlPR10 promoter ( pSlPR10 ) was mainly detected in the exocarp of transgenic tomato plants of both Ailsa Craig and Micro-Tom cultivars. This promoter was further utilized for transgenic expression of SlANT1 and SlMYB31 in tomato, which are master regulators of anthocyanin and cuticular wax biosynthesis, respectively. pSlPR10 -driven SlANT1 expression resulted in anthocyanin accumulation in the exocarp, conferring gray mold resistance and extended shelf life to the fruit, while SlMYB31 expression led to waxy thickening in the fruit skin, delaying water loss and also extending fruit shelf life. Intriguingly, pSlPR10 and two other weaker tomato exocarp-preferential promoters exhibited coincided expression specificities in the gynophore of transgenic Arabidopsis ( Arabidopsis thaliana ) plants, providing not only an inkling of evolutionary homology between tomato exocarp and Arabidopsis gynophore but also useful promoters for studying gynophore biology in Arabidopsis . Collectively, this work reports a desirable promoter enabling targeted gene expression in tomato exocarp and Arabidopsis gynophore and demonstrates its usefulness in genetic improvement of tomato fruit quality., Competing Interests: Based on the data of the current research, a China invention patent (ZL202210684553.8) has been granted to the authors., (© The Author(s) 2024. Published by Oxford University Press on behalf of Nanjing Agricultural University.)

Base editors (BEs) empower the efficient installation of beneficial or corrective point mutations in crop and human genomes. However, conventional BEs can induce unpredictable guide RNA (gRNA)-independent off-target edits in the genome and transcriptome due to spurious activities of BE-enclosing deaminases, and current improvements mostly rely on deaminase-specific mutagenesis or exogenous regulators. Here we developed a split deaminase for safe editing (SAFE) system applicable to BEs containing distinct cytidine or adenosine deaminases, with no need of external regulators. In SAFE, a BE was properly split at a deaminase domain embedded inside a Cas9 nickase, simultaneously fragmenting and deactivating both the deaminase and the Cas9 nickase. The gRNA-conditioned BE reassembly conferred robust on-target editing in plant, human and yeast cells, while minimizing both gRNA-independent and gRNA-dependent off-target DNA/RNA edits. SAFE also substantially increased product purity by eliminating indels. Altogether, SAFE provides a generalizable solution for BEs to suppress off-target editing and improve on-target performance., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)

Objective: To investigate the clinical features of children with chronic nonbacterial osteomyelitis (CNO), and raise awareness among clinicians. Methods: In this retrospective study, 18 patients with CNO who were diagnosed in Children's Hospital of Fudan University from January 2015 to December 2021 were included. Results: Eighteen children with CNO (12 males, 6 females) were identified. Their age at onset was 9 (5, 11) years, the delay in diagnosis was 2 (1, 6) months, and follow-up-was 17 (8, 34) months. The most common symptoms were fever in 14 children, as well as bone pain and (or) arthralgia in 14 children. In terms of laboratory results, normal white blood cell counts were observed at onset in 17 patients; increased erythrocyte sedimentation rate (ESR) in all patients; increased C reactive protein (CRP) over the normal value in 14 patients. Of the 18 patients, 2 had positive antinuclear antibodies, while none had positive human leukocyte antigen-B27 or rheumatoid factor. Imaging examination revealed that all the patients had symmetrical and multifocal skeletal lesions. The number of structural lesions detected by imaging investigation was 8 (6, 11). The most frequently affected bones were tibia in 18 patients and femur in 17 patients. Bone biopsy was conducted in 14 patients and acute or chronic osteomyelitis manifested with inflammatory cells infiltration were detected. Magnetic resonance imaging (MRI) found bone lesions in all the patients and bone scintigraphy were positive in 13 patients. All the patients were treated with nonsteroidal anti-inflammatory drugs, among whom 10 cases also treated with oral glucocorticoids, 9 cases with traditional disease modifying anti-rheumatic drugs, 8 cases with bisphosphonates and 6 cases with tumor necrosis factor inhibitors. The pediatric chronic nonbacterial osteomyelitis disease activity score, increased by 70% or more in 13 patients within the initial 6-month follow-up. Conclusions: The clinical manifestations of CNO are lack of specificity. The first symptom of CNO is fever, with or without bone pain and (or) arthralgia, with normal peripheral blood leukocytes, elevated CRP and (or) ESR. Whole body bone scanning combined with MRI can early detect osteomyelitis at subclinical sites, and improve the diagnostic rate of CNO.

Objective: To investigate A1 pulley thickness of flexor tendon in healthy volunteers and to analyze its influence factors., Methods: The study included 90 healthy volunteers and the A1 pulley thickness at bilateral fingers was measured using high frequency ultrasound. The following parameters were recorded for each participant: age, gender, weight, height, body mass index (BMI)., Results: High-frequency ultrasound can clearly show A1 pulley. There was no significant difference in A1 pulley thickness between the bilateral fingers ( P >0.05). A1 pulley thickness was significantly different in different fingers ( P <0.05). Further comparison showed that A1 pulley thickness could be divided into two subsets: thumb and little finger ((0.196±0.051) mm), index, middle and ring fingers ((0.230±0.055) mm). A1 pulley thickness was positively correlated with age ( r =0.468, P <0.001). The normal reference ranges for thumb and little finger were 0.09-0.23 mm, 0.12-0.30 mm and 0.12-0.32 mm, respectively. The normal reference ranges for index, middle and ring fingers were 0.11-0.27 mm, 0.15-0.35 mm and 0.17-0.35 mm in volunteers aged 3-19 yr., 20-49 yr., and ≥50 yr., respectively. Gender and BMI had negligible impact on A1 pulley thickness ( P >0.05)., Conclusion: High-frequency ultrasound can clearly show and measure A1 pulley. Site and age should be taken into account when determining the reference range of normal A1 pulley thickness. High-frequency ultrasound can be a quantitative evaluation method for A1 pulley lesions., (Copyright© by Editorial Board of Journal of Sichuan University (Medical Sciences).)

Main Conclusion: In Hordeum vulgare, nine differentially expressed novel miRNAs were induced by colchicine. Five novel miRNA in colchicine solution showed the opposite expression patterns as those in water. Colchicine is a commonly used agent for plant chromosome set doubling. MicroRNA-mediated responses to colchicine treatment in plants have not been characterized. Here, we characterized new microRNAs induced by colchicine treatment in Hordeum vulgare using high-throughput sequencing. Our results showed that 39 differentially expressed miRNAs were affected by water treatment, including 34 novel miRNAs and 5 known miRNAs; 42 miRNAs, including 37 novel miRNAs and 5 known miRNAs, were synergistically affected by colchicine and water, and 9 differentially expressed novel miRNAs were induced by colchicine. The novel&#95;mir69, novel&#95;mir57, novel&#95;mir75, novel&#95;mir38, and novel&#95;mir56 in colchicine treatment showed the opposite expression patterns as those in water. By analyzing these 9 differentially expressed novel miRNAs and their targets, we found that novel&#95;mir69, novel&#95;mir56 and novel&#95;mir25 co-target the genes involving the DNA repair pathway. Based on our results, microRNA-target regulation network under colchicine treatment was proposed, which involves actin, cell cycle regulation, cell wall synthesis, and the regulation of oxidative stress. Overall, the results demonstrated the critical role of microRNAs mediated responses to colchicine treatment in plants.

Pulmonary arterial hypertension (PAH) is a chronic progressive disease which leads to elevated pulmonary arterial pressure and right heart failure. 3,7-Bis(2-hydroxyethyl)icaritin (ICT), an icariin derivatives, was reported to have potent inhibitory activity on phosphodiesterase type 5 (PDE5) which plays a crucial role in the pathogenesis of PAH. The present study was designed to investigate the effects of ICT on monocrotaline (MCT)-induced PAH rat model and reveal the underlying mechanism. MCT-induced PAH rat models were established with intragastric administration of ICT (10, 20, 40 mg/kg/d), Icariin (ICA) (40 mg/kg/d) and Sildenafil (25 mg/kg/d). The mean pulmonary arterial pressure (mPAP) and right ventricle hypertrophy index (RVHI) were measured. Pulmonary artery remodeling was assessed by H&E staining. Blood and lung tissue were collected to evaluate the level of endothelin 1 (ET-1), nitric oxide (NO), and cyclic guanosine monophosphate (cGMP). The expressions endothelial nitric oxide synthase (eNOS) and PDE5A in lung tissues were determined by Western blot analysis. The results showed that ICT reduced RVHI and mPAP, and reversed lung vascular remodeling in rats with MCT-induced PAH. ICT also reversed MCT-induced ET-1 elevation, NO and cGMP reduction in serum or lung tissue. Moreover, ICT administration significantly induced eNOS activation and PDE5A inhibition. ICT with lower dose had better effects than ICA. In summary, ICT is more effective in preventing MCT-induced PAH in rats via NO/cGMP activation compared with ICA. These findings demonstrate a novel mechanism of the action of ICT that may have value in prevention of PAH., (Copyright © 2018 Elsevier B.V. All rights reserved.)

To provide scientific basis for reasonable application of nitrogen and create varieties with high N use-efficiency, an experiment was carried out to study the effects of nitrogen fertilizer application rate on grain yield, N use rate and quality of different rice varieties. Four different genotypic rice varieties, Nipponbare, N70, N178 and OM052 were used as tested material and three levels of nitrogen application rate (0, 120, 270 kg·hm -2 ) were conducted. Urea as nitrogen source was applied as basal (70%) and panicle (30%) fertilizer. The results showed that nitrogen fertilizer could raise yield mainly because of the increased effective panicles and filled grains per panicle. When the N application rate was 120 and 270 kg·hm -2 , OM052 had the largest grain yield among four varieties, being 41.1% and 76.8% higher, respectively compared with control. Difference in grain yield among four varieties was due to the difference of nitrogen use efficiency. Under 120 and 270 kg·hm -2 nitrogen levels, Nipponbare had the lowest grain yield and N agronomic efficiency (NAE, 40.90 g·g -1 and 18.56 g·g -1 ), which was a variety with low N use-efficiency. On the contrary, OM052 had the highest grain yield and NAE (145.9 g·g -1 and 81.24 g·g -1 ), was a variety with high N use-efficiency. N fertilizer application increased the amylose content and protein content, lengthened gel consistency, reduced chalky kernel, chalkiness, and alkali digestion value. With the increase of N fertilizer application, hot paste viscosity, peak viscosity, consistence viscosity and breakdown viscosity were decreased gradually, and setback viscosity was increased. Correlation analysis showed that the yield and yield components had more significant correlations with appearance quality, cooking and eating quality under low N level. This study confirmed that OM052 was a double high variety with extremely high N agronomic efficiency and yield. Reasonable application of nitrogen fertilizer could significantly increase effective panicles and filled grains per panicle, improve rice quality, and ensure high yield and superior quality simultaneously.

Ethnopharmacological Relevance: Honeysuckle (Lonicera japonica Thunb.), a traditional Chinese herb, has widely been used to treat pathogen infection. However, the underlying-mechanism remains elusive., Aims of the Study: To reveal the host microRNA (miRNA) profile with the anti-viral activity after honeysuckle treatment., Materials and Methods: Here we reveal the differentially expressed miRNAs by Solexa ® deep sequencing from the blood of human and mice after the aqueous extract treatment. Among these overexpressed innate miRNAs both in human and mice, let-7a is able to target the NS1 region (nt 3313-3330) of dengue virus (DENV) serotypes 1, 2 and 4 predicated by the target predication software., Results: We confirmed that let-7a could target DENV2 at the predicated NS1 sequence and suppress DENV2 replication demonstrated by luciferase-reporter activity, RT-PCR, real-time PCR, Western blotting and plaque assay. ICR-suckling mice consumed honeysuckle aqueous extract either before or after intracranial injection with DENV2 showed decreased levels of NS1 RNA and protein expression accompanied with alleviated disease symptoms, decreased virus load, and prolonged survival time. Similar results were observed when DENV2-infected mice were intracranially injected with let-7a., Conclusion: We reveal that honeysuckle attenuates DENV replication and related pathogenesis in vivo through induction of let-7a expression. This study opens a new direction for prevention and treatment of DENV infection through induction of the innate miRNA let-7a by honeysuckle., (Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.)

Percutaneous coronary intervention (PCI) is widely used in clinical treatment of coronary artery disease. However, the effects of PCI on preventing restenosis after revascularization and improving the quality of life were not satisfying. Huxin Formula is formulated by modifying an experienced Chinese medicine formula and has been widely used in clinical practice due to its marked effects on coronary heart disease. A multicentre double-blind randomized controlled clinical trial was designed to evaluate the effects and safety of Huxin Formula in patients undergoing PCI. Our results showed that there was no significant difference between the two groups in main outcomes. For patients with ejection fraction (EF) >50%, score of the quality of life scale was higher in treatment group compared with control group. For patients with unstable angina, score of the quality of life scale in 360 days was significantly higher in treatment group compared with control group (P < 0.05). No obvious adverse reaction was found in the use of Huxin Formula. In conclusion, Huxin Formula, believed to be a safe treatment for patients after PCI, has benefits in improving the quality of life in patients with unstable angina though it failed to show superiority in primary and secondary outcomes.

Background: Coronary artery bypass grafting (CABG) is widely used in the treatment of coronary artery disease. A multicenter, double-blind, randomized, controlled clinical trial was designed to evaluate the efficacy and safety of Huxin Formula post CABG., Patients and Methods: 270 inpatients with coronary heart disease participated in this study. CABG patients in the control group were treated with placebo, while patients in the experimental group were treated with Huxin Formula 1 week after the surgery. All patients were treated for 6 months and followed up for another 6 months. The main outcomes (death, nonfatal myocardial infarction, repeat revascularization, and readmission) were assessed 360 days after treatment, and secondary outcomes (frequency and scores of angina pectoris, etc.) were assessed 0, 90, 180, 270, and 360 days after treatment., Results: Our results showed no significant difference between the 2 groups for the primary endpoints. In patients with cardiac function class II (New York Heart Association), the score of angina pectoris was significantly lower (3.88 ± 3.86 vs. 5.45 ± 3.59) and the frequency of angina pectoris attacks was less (0.96 ± 1.01 vs. 1.36 ± 0.94) after 90 days of treatment with Huxin Formula compared to placebo (p < 0.05). In patients with 3 coronary vessel lesions, the cardiac function class (1.14 ± 0.35 vs. 1.05 ± 0.21) after 360 days was significantly higher in the control group compared to the treatment group (p < 0.05). There were no obvious adverse reactions., Conclusion: Huxin Formula may improve cardiac function of patients with 3 coronary vessel lesions and relieve symptoms of patients with cardiac function class II but failed to show superiority in primary outcomes., (© 2015 S. Karger GmbH, Freiburg.)

Aquaporins are thought to be associated with water transport and play important roles in cotton (Gossypium hirsutum) fibre elongation. Among aquaporins, plasma membrane intrinsic proteins (PIPs) constitute a plasma-membrane-specific subfamily and are further subdivided into PIP1 and PIP2 groups. In this study, four fibre-preferential GhPIP2 genes were functionally characterized. The selective interactions among GhPIP2s and their interaction proteins were studied in detail to elucidate the molecular mechanism of cotton fibre development. GhPIP2;3 interacted with GhPIP2;4 and GhPIP2;6, but GhPIP2;6 did not interact with GhPIP2;4. Coexpression of GhPIP2;3/2;4 or GhPIP2;3/2;6 resulted in a positive cooperative effect which increased the permeability coefficient of oocytes, while GhPIP2;4/2;6 did not. GhBCP2 (a blue copper-binding protein) inhibited GhPIP2;6 water channel activity through their interaction. Overexpression of GhPIP2 genes in yeast induced longitudinal growth of the host cells. By contrast, knockdown of expression of GhPIP2 genes in cotton by RNA interference markedly hindered fibre elongation. In conclusion, GhPIP2 proteins are the primary aquaporin isoforms in fibres. They selectively form hetero-oligomers in order to regulate their activities to meet the requirements for rapid fibre elongation., (© 2013 The Authors. New Phytologist © 2013 New Phytologist Trust.)

Objective: To observe the effect of Huxin Formula on expressions of the chief reverse cholesterol transport (RCT) associated genes, caveolin-1 and scavenger receptor-BI (SR-BI) in ApoE-gene knockout [ApoE (-/-)] mice., Methods: Thirty ApoE (-/-) mice of 4-6 weeks old were randomly divided into three groups (A-C). After being fed with high-fat diet for 16 weeks, they were treated with HXF (1 mL/100 g), pravachol (0.3 mg/100 g), and saline in equal volume respectively for 16 weeks successively; in addition, a blank group was set up with 10 C57BL/6J mice of 6-week old received 16-week high-fat feeding and saline treatment. Animals were sacrificed at the termination of the experiment, their paraffin sections of aortic tissue were used to measure the size of plaque, expressions of cavolin-1 and SR-BI were detected by immunological histochemical method., Results: As compared with the blank group, levels of caveolin-1 and SR-BI were increased in Groups A and B (P<0.01); but the increase in Group A was more significant than that in Group B (P<0.05). The plaque/aorta area ratio decreased significantly in Groups A and B, but showed insignificant difference between the two groups., Conclusion: HXF could obviously increase the expressions of RCT associated genes, caveolin-1 and SR-BI, promote the RCT process, so as to reduce the formation of aorta atherosclerotic plaque in ApoE (-/-) mice.

A series of salicylanilide derivatives (compounds 1-32) were synthesised by reacting substituted salicylic acids and anilines. The chemical structures of these compounds were determined by (1)H-NMR, electrospray ionisation mass spectrometry (ESI-MS) and elemental analysis. The compounds were assayed for their antiproliferative activities against the Hep-G2 cell line by the 3-(4,5-dimethylthylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Among the compounds tested, 22 and 28 showed the most favouable antiproliferative activities with 50% inhibitory concentration (IC(50)) values of 1.7 and 1.3 μM, respectively, which were comparable to the positive control of 5-fluorouracil (IC(50)=1.8 μM). A solid-phase ELISA assay was also performed to evaluate the ability of compounds 1-32 to inhibit the autophosphorylation of the epidermal growth factor receptor tyrosine kinase (EGFR TK). Docking simulations of 22 and 28 were carried out to illustrate the binding mode of the molecule into the EGFR active site, and the result suggested that both compounds 22 and 28 could bind the EGFR kinase well.

Copper is vitally required for plants at low concentrations but extremely toxic for plants at elevated concentrations. Plants have evolved a series of mechanisms to prevent the consequences of the excess or deficit of copper. These mechanisms require copper-interacting proteins involved in copper trafficking. Blue copper-binding proteins (BCPs) are a class of copper proteins containing one blue copper-binding domain binding a single type I copper. To investigate the role of BCPs in plant development and in response to stresses, we isolated nine cDNAs encoding the putative blue copper-binding proteins (GhBCPs) from cotton (Gossypium hirsutum). Meanwhile, four corresponding genes (including GhBCP1-GhBCP4), which contain a single intron inserted in their conserved position, were isolated from cotton genome. Quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis indicated that the nine GhBCP genes are differentially expressed in cotton tissues. Among them, GhBCP1 and GhBCP4 were predominantly expressed in fibers, while the transcripts of GhBCP2 and GhBCP3 were accumulated at relatively high levels in fibers. These four genes were strongly expressed in early fiber elongation, but dramatically declined with further fiber development. In addition, these GhBCP genes were upregulated in fibers by Cu(2+) , Zn(2+) , high-salinity and drought stresses, but downregulated in fibers by Al(3+) treatment. Overexpression of GhBCP1 and GhBCP4 in yeast (Schizosaccharomyces pombe) significantly increased the cell growth rate under Cu(2+) , Zn(2+) and high-salinity stresses. These results suggested that these GhBCPs may participate in the regulation of fiber development and in response to high-salinity and heavy metal stresses in cotton., (Copyright © Physiologia Plantarum 2010.)

Objectives: The emergence of antibiotic-resistant Helicobacter pylori strains has necessitated a search for alternative therapies for the treatment of this infection. The aim of this study was to evaluate whether or not polysaccharide fractions from Aloe vera are effective in inhibiting the adherence of H. pylori in vitro., Methods: Polysaccharide fractions were extracted from A. vera and subjected to carbohydrate analysis. The adhesive effect was determined by co-incubation of H. pylori and cells with polysaccharides followed by fluorescein isothiocyanate labelling and Gram staining in vitro. Inhibition of H. pylori growth and cellular viability was tested by agar diffusion and MTT assay., Key Findings: APS-F2 contained significant amounts of galacturonic acid, galactose and arabinose. APS-F1 was galacturonic acid-free and consisted of mannose, glucose and galactose. APS-F2 (0.1, 0.5 and 1.0 mg/ml) reduced the count of H. pylori attached to MKN45 cells to 88, 76 and 64%, respectively. APS-F1 did not show the same effect. Neither polysaccharide revealed an inhibitory effect on the growth of H. pylori or cell viability. In addition, APS-F2 was shown to have a potent anti-adhesive effect against Escherichia coli., Conclusions: The results show that the acidic polysaccharide from A. vera has a potent anti-adhesive effect against H. pylori in vitro. However, there have yet to be any in-vivo studies to demonstrate the clinical relevance of this finding., (© 2010 The Authors. JPP © 2010 Royal Pharmaceutical Society of Great Britain.)

We sought to evaluate the impact of C-reactive protein (CRP) levels on in-stent restenosis after percutaneous coronary intervention.The plasma level of CRP is considered a risk predictor for cardiovascular diseases. However, the relationship between CRP and in-stent restenosis has been a matter of controversy. Meta-analysis reduces variability and better evaluates the correlation.We performed a systemic search for literature published in March 2008 and earlier, using MEDLINE(R), the Cochrane clinical trials database, and EMBASE(R). We also scanned relevant reference lists and hand-searched all review articles or abstracts from conference reports on this topic. Of the 245 studies that we initially searched, we chose 9 prospective observational studies (1,062 patients).Overall, CRP concentration was higher in patients who experienced in-stent restenosis. The weighted mean difference in CRP levels between the patients with in-stent restenosis and those without was 1.67, and the Z-score for overall effect was 2.12 (P=0.03). Our subgroup analysis that compared patients with stable and unstable angina showed a weighted mean difference in the CRP levels of 2.22 between the patients with and without in-stent restenosis, and the Z-score for overall effect was 2.23 (P=0.03) in 5 studies of unstable-angina patients. There was no significance in 4 studies of stable-angina patients.In spite of significant heterogeneity across the studies, our meta-analysis suggests that preprocedurally elevated levels of CRP are associated with greater in-stent restenosis after stenting and that this impact appears more prominent in unstable-angina patients.

Objective: To investigate the inclusion technology conditions of volatile oil extracted from Herba Pogostemonis and Exocarpium citri in Huxin Fang granules by beta-cyclodextrin., Methods: The orthogonal design was used. The percent conversion of the oil in inclusion and the yield of inclusion complex were the indices to determine the factors containing the ratio of the oil to beta-cyclodextrin, the ratio of beta-cyclodextrin to water and the grinding time of the mixture of the oil, beta-cyclodextrin and water., Results: The optimum conditions for the inclusion were as follows: the ratio of the oil to beta-cyclodextrin was 1:10, the ratio of beta-cyclodextrin to water was 1:3, and the grinding time was 40 min., Conclusion: The optimum conditions for the inclusion were steady and feasible.

Cotton (Gossypium hirsutum), the most important textile crop worldwide, often encounters water stress such as drought or waterlog during its growth season (Summer). To investigate molecular mechanism of water regulation in cotton plants, three cDNAs encoding the plasma membrane intrinsic protein (PIP) were isolated from cotton root cDNA library, and designated GhPIP1;1, GhPIP2;1 and GhPIP2;2, respectively. All of the three PIP proteins displayed water channel activity in Xenopus laevis oocytes. GhPIP2;1 and GhPIP2;2 proteins, however, showed much higher water transport activity than that of the GhPIP1;1 protein. Northern blot analysis revealed that all of the three genes were preferentially expressed in young roots. Further analysis by Real-time quantitative RT-PCR revealed that the transcripts of all the three genes were accumulated at high levels in 3-day-old young roots, but dramatically declined to much lower levels in 6-14 days old roots during seedling development, suggesting that expressions of the isolated GhPIP genes are developmentally regulated in roots. Additionally, expressions of the three genes were remarkably up-regulated or down-regulated under different stresses such as NaCl, cold, PEG (polyethylene glycol) treatments. Collectively, the results suggest that these genes may be involved in root development and in response to stresses.

Objective: To evaluate the randomized controlled trials on preventing and treating restenosis after percutaneous coronary intervention (PCI) with Chinese medicine adopting the principle of evidence-based medicine (EBM)., Methods: Pertinent literatures were retrieved roundly, and scored depending on Jadad Quality Scale. Meta-analysis was applied on the total effect of outcomes reported using RevMan 4. 2 software. Moreover, the sensitivity was analyzed, the publication bias was identified with funnel-plot and the adverse reaction of medicine was reported., Results: Among the 17 literatures met the inclusion criteria, only two double-blinded trials were appraised as high quality. The analysis on the recurrence rates of restenosis post-PCI and angina pectoris in patients treated with Chinese medicine illustrated that the rates in the treatment group were all lower than those in the control group with statistical significance (P < 0.01). Sensitivity analysis revealed a relatively stable result of the evaluation. Funnel-plot displayed an asymmetrical figure, indicating the possible existence of publication bias., Conclusion: TCM has definite therapeutic effect on preventing and treating restenosis after PCI and reducing the recurrence rate of angina pectoris in patients.

Application of traditional Chinese medicine and drugs in cardiac surgery have won great progress in recent 10 years, mainly presented in aspects of the treatment of coronary heart disease, myocardium protection, cardiopulmonary bypass and pulmonary hypertension.

Objective: To evaluate the clinic effect of abdominal needling (AN) on post-cardiosurgical operational gastrointestinal dysfunction., Methods: Sixty patients were equally assigned to two groups, the AN group and the control group. AN applied on the AN group was conducted by needling at Xiawan (Ren 10), Qihai (Ren 6), Qipang, Guanyuan (Ren 4), Qixue, Shuifen (Ren 9), Tianshu (St. 25) and Daju (St. 27), starting from the first day after operation, once every day with the needle retained time of 20 min, 5 days as one therapeutic course. The control group was treated with the conventional post-operational measurement with intravenous injection of 20 mg Omeprazole once a day, for 3 successive days. The score on symptoms as anorexia, weakness, abdominal distension, constipation, laziness to speak, nausea and vomiting, weakened borborygmus, pale-yellow complexion and edema, etc. , were observed., Results: The symptom scores in the aspects of anorexia, weakness, abdominal distension, constipation, weakened borborygmus, nausea and vomiting in the AN group were significant difference when compared with before treatment and the control group (P <0.01). After treatment the improvement in the control group only showed in weakness and pale-yellow complexion (P <0.01). Observation on gastric emptying showed that 24 h gastric paresis occurred in 17 patients (56.7%) of the AN group, and 22 (73.3%) of the control group; the 24 h and 72 h barium residue (strip) in the AN group was 10.60 +/- 4.61 and 8.53 +/- 4.08 respectively, while in the control group 17.07 +/- 3.99 and 15.83 +/- 4.19; the 24 h, 72 h gastric emptying rate in the former was (47.00 +/- 23.07) % and (57.33 +/- 20.37) %, and in the latter (14.67 +/- 19.95) % and (20.83 +/- 20.97) %, all showed significant difference between them (P <0.05 or P <0.01). Levels of motilin in the two groups after operation were decreased showing significant difference in comparison with before operation (P < 0.01), and AN group was raised after teratment, also showing significant difference between groups (P <0.01)., Conclusion: AN can strengthen gastrointestinal peristalsis, promote gastrointestinal emptying, recover quickly gastrointestinal hormone to the normal range. Besides, it is safe and effective, with no pain or only slight pain, easy in locating the acupoint accurately, standard prescription, high compliance of patients, and convenient for clinical operating, therefore, it is worthy of spreading in clinical practice.

An improved and simple method using capillary gas chromatography coupled with a flame ionization detection system (GC-FID) has been developed for quantitative analysis of free and conjugated phytosterols in tobacco. Direct acid and alkaline hydrolysis were first introduced into tobacco analysis to liberate free phytosterols from conjugates, followed by extraction with hexane, derivatization to trimethylsilyl ether derivatives and finally GC quantitative determination. The generality and applicability of this improved method for analyzing free and conjugated phytosterols in tobacco were validated after a series of optimization and comparison were done. Compared with traditional methods, this improved method not only simplified procedures, but also saved time and solvent. The limits of detection (LODs) of phytosterols varied from 0.35 to 0.10 microg mL(-1), the relative standard deviations (RSD) were from 2.3% to 3.3% and recovery ranged from 87% to 99%. The analysis results showed that total phytosterols' content in tobacco ranged from 1.0 to 2.5 mg g(-1), and most phytosterols existed as conjugates, only approximately 15-25% phytosterols existed in free-form. Ergosterol was only found in mildewy flued-cured tobacco and the level was approximately 0.2-0.25 mg g(-1).

Objective: To explore the diagnostic figures for TCM syndrome typing in coronary heart disease (CHD) patients., Methods: A retrospective investigation was carried out in 319 CHD patients hospitalized from Jan. 2004 to Dec. 2004 in authors' hospital. Through cluster analysis, descriptive statistics and frequency normalization in combination of clinical observation, the diagnostic figures of TCM syndromes were obtained., Results: The figures for qi deficiency syndrome were: primary symptoms: chest pain and stuffiness, secondary symptoms: tiredness, short breath, poor appetite, light colored tongue, deep and thready pulse; for qi deficiency with phlegm and blood stasis syndrome: primary symptoms: chest stuffiness and pain, secondary symptoms: tiredness, insomnia, palpitation, obesity, dark red tongue, string and slippery pulse; for turbid-phlegm blocking collateral syndrome: primary symptoms: chest stuffiness, secondary symptoms: cough, expectoration with much white sputum, tiredness, short breath and poor appetite, light colored tongue with white greasy coating, slippery pulse., Conclusion: Research on diagnostic criteria for TCM syndrome typing could be established upon clinical epidemiologic survey and statistic analysis in combining with specialists' suggestions to primarily set the referrence figures.

Objective: To study TCM syndrome distribution laws in patients with coronary heart diseases (CHD) by epidemiological investigation., Methods: A clinical survey was carried out in 319 inpatients with CHD, whose diagnosis was confirmed by coronary arteriography, in the authors' hospital from January 2004 to December 2004. The TCM syndrome distribution laws were analyzed, and the relationship of coronary arteriographic picture with TCM syndrome elements, common symptoms, pulse and tongue figures, as well as the correlation between syndrome typing and blood-lipid levels were analyzed, too., Results: Qi deficiency was the most popular syndrome in patients with CHD (87.1%), blood stasis syndrome and phlegm retention syndrome took the second place, accounting for 79.9% and 78.7% respectively. No significant difference was shown in comparison of tongue and pulse figures with the affected branches of coronary artery, the dark-pale tongue with white greasy fur and taut-slippery pulse being the dominance in patients. The blood-lipid levels in patients with various TCM syndrome types were similar, showing insignificant difference., Conclusion: The TCM pathogenesis of CHD takes qi deficiency as the core, blood stasis and phlegm retention as the important pathologic products.

Objective: To observe the effects of Shenfu Injection (SI) on immune function in patients during perioperative cardiopulmonary bypass (CPB)., Methods: Ninety patients scheduled for CPB were randomly divided into 3 groups, the group A treated with SI before CPB, the group B treated with SI after CPB and the group C (the control group), 30 cases in each group. Counts of leukocyte, neutrophil, lymphocyte and T lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+) and serum levels of IgG, IgA, IgM, C3 and C4 were investigated at different time-points, i.e. one day before CPB, immediately after anesthesia and finishing CPB, and the 1st, 3rd and 8th post-operational days., Results: In all the three groups, counts of leukocyte and neutrophil rose immediately after CPB and the high level maintained on the 1st and 3rd post operational days, it somewhat lowered on the 8th day but still higher than that before operation, showing insignificant difference among the 3 groups. As for the other indexes, they declined obviously and immediately after CPB and began to restore on the 8th day to a certain extent, and the restoration were better in the group A and B, as the changes were milder., Conclusion: SI could improve the immune function and reduce the risk of post operational infection in patients during perioperative CPB.

Objective: To evaluate the clinical efficacy of treatment for regulating Pi and protecting Xin (abbr. as CHM) and its effect on quality of life (QOL) in patients after coronary artery bypass grafting (CABG)., Methods: One hundred and six patients, who were planned to undergo CABG were assigned into two groups, the 51 patients in the control group were treated with conventional Western medicinal treatment (WM) and the 55 patients in the experimental group were treated by WM with additional medication of CHM constituted mainly by modified Huxin Recipe. The changes of symptoms and heart function were observed and the patients' QOL was evaluated by SF-36 Scale., Results: After being treated for 3 months, the total scores of symptoms in the experimental group was significantly improved in comparing with that in the control group (P < 0.01), the clinical efficacy on symptoms of TCM and improvement of heart function were superior to those in the control group (P < 0.05) respectively. The scores in terms of pain, vitality, role of emotion, mental health, health transition of patients in the experimental group according to SF-36 scale were significantly higher than those in the control group (P < 0.05 or P < 0.01)., Conclusion: Treatment for regulating Pi and protecting Xin by modified Huxin Recipe can improve the recovery process of patients after CABG, elevate patients' heart function, symptoms and QOL.

Objective: To evaluate the clinical effect and quality of life (QOL) in coronary artery bypass grafting (CABG) patients treated according to syndrome differentiation of TCM., Methods: Eighty-two CABG patients were allocated in the control group (n = 39) treated with conventional method of western medicine, and the TCM-treated group (n = 43) treated according to syndrome differentiation of TCM based on the conventional western medicine. The Seattle Angina Questionnaire (SAQ) was used to assess the clinical effect and quality of life., Results: After 2 months treatment, the TCM syndrome score in TCM-treated group was better, and the score of SAQ was higher than that in the control group (all P < 0.01). In the aspects of anginal stability, anginal frequency, treatment satisfaction and disease perception, there was significant difference between the two groups (P < 0.05 or P < 0.01)., Conclusion: TCM treatment could promote recovery, improve symptoms and elevate QOL of CABG patients.

Objective: To explore the patterns of Syndrome Differentiation of coronary artery bypass graft (CABG) on the peri-operative coronary heart disease (CHD) patients., Methods: One week after operation, thirty-seven CHD patients, who received CABG of internal mammary artery or great saphenous vein under conventional general anesthesia with low or middle temperature extracorporeal circulation were differentiated as various syndromes, the pre- and post-operational ECG, color Doppler echocardiography as well as during and after operation. The hemodynamic parameters were monitored., Results: In the CHD patients, 64.9% were differentiated as Qi-Yin deficiency, 67.6% were complicated with Phlegm Syndrome and 62.2% with Stasis in blood, suggesting that Qi-deficiency, phlegm and stasis are the basic pathogenesis in patients after CABG. Moreover, the peri-operative Syndrome was correlated with the condition of coronary arterial lesions, heart and lung functions before operation, and the time for extracorporeal circulation during the operation., Conclusion: TCM Syndrome Differentiation conducting in peri-operative stage might be useful to explore the patterns of Syndrome alteration which provided a basis for preventing peri-operational complication and elevating success rate of operation.

Objective: Nitric oxide has been reported to reduce intimal hyperplasia as a response to arterial injury. This study was designed to assess the possible effect of perivascular application of a nitric oxide donor on neointimal proliferation occurring in veins exposed to the dynamics of the arterial circulation in a hypercholesterolemic rabbit model., Methods: Autologous jugular vein grafts were implanted in the carotid circulation of 20 hypercholesterolemic rabbits. A mixture of a biodegradable polymer and the nitric oxide donor, spermine/nitric oxide, which releases nitric oxide with a half-life of 39 minutes, was applied periadventitially at the time of implantation. Controls were veins bathed in saline solution, polymer alone, and polymer plus the carrier vehicle spermine without nitric oxide. Animals (n = 5 in each group) were put to death on day 28 for morphometric analysis, cell count, and immunohistochemical staining., Results: Treatment with perivascular nitric oxide donor significantly decreased wall thickness (126 +/- 24 microm vs 208 +/- 45 microm, p = 0.0017) and area (124 +/- 22 microm2/microm vs 211 +/- 37 microm2/microm, p = 0.005). With the carrier vehicle spermine alone, there was a trend toward reduced intimal thickness, but the change was not statistically significant. In the grafts treated with nitric oxide donor, expression of insulin-like growth factor, fibroblast growth factor, thrombospondins, fibronectin, and tenascin was reduced., Conclusion: The periadventitial delivery of nitric oxide donor produces a reduction of neointimal hyperplasia in veins implanted in the arterial circulation. The mechanism of action is not entirely clear, but the reduction cannot be explained on the basis of decreased cell proliferation alone. Other possibilities are modulation of protein synthesis of vascular smooth muscle cells and production of extracellular matrix components.

Flow visualization and pressure measurements were carried out in a singel valve saphenous vein casting which was made from a saphenous vein segment obtained from a bypass patient at Cedars Sinai Medical Center. Dye was injected to understand the flow around the valve. The dye showed very complex flow patterns around the valve and in the valve sinus, and the cavity formed by a ligated branch. For steady flow, pressure drops across the valve were 0.72, 2.0 and 6.3 mmHg for the physiological flow rates of 45, 84, and 169 ml/min, respectively. Overall pressure drop across the casting (compared to Poiseuille flow for a straight tube) increased with the flow rate, being 130 to 290 percent higher over this flow rate range. In the case of pulsatile flow, pressure drops across the valve were 0.95 and 3.0 mmHg for the flow rates of 47 and 87 ml/min which were 26 and 43 percent higher than those of steady flow. Overall pressure drop was 220 and 360 percent higher for those flow rates compared to Poiseuille flow. The measured spatial pressure distributions along the casting and flow visualization indicated the global nature of the flow field with the accelerated flow through the valve separating and reattaching downstream along the wall in the pressure recovery region. Atherosclerosis may be prone to occur in the lower shear region along the wall beyond the valve tip in the reattachment region, as we have observed in vivo in rabbit experiments.

Coronary saphenous vein grafts in human beings have a more limited long-term patency rate than internal thoracic artery grafts, primarily because of more rapid development of arteriosclerosis. The factors responsible for this increased susceptibility are not completely understood. To test the hypothesis that vein valves may influence this process, we studied 48 hypercholesterolemic rabbits with jugular vein grafts interposed into the carotid arterial circulation. In 24 animals (group A), the vein segments did not contain a vein valve. In the other 24 animals (group B), a vein valve was present. Both groups were further divided in four subgroups of six to be put to death at 2, 4, 6, and 8 weeks after the operation. All animals were fed a 2% cholesterol diet. At postmortem examination, alternate 2 mm sections were either stained with hematoxylin and eosin for histologic and morphometric studies or frozen in liquid nitrogen for immunohistochemistry and in situ hybridization studies. Proliferating cell nuclear antigen immunostaining was used to study cell proliferation. Wall thickness of vein grafts increased with time. During the first 2 weeks intimal and medial thickening was primarily due to an increase in numbers of cells. Between 2 and 6 weeks further intimal and medial thickening occurred, but without additional increase in cell numbers. After 6 weeks, foam cells and lipid deposits started to appear. By 8 weeks, changes identical to those seen in arteriosclerotic plaques in human beings were evident. These changes developed sooner and with more intensity in group B animals (p < 0.01 to 0.001), and they developed faster and with more severity in segments of vein located distal to the valve than in the segments located proximal to the valve (p < 0.001). This is the first controlled experiment demonstrating that the presence of valves in the vein segments is associated with augmented and accelerated intimal changes leading to vein atheromatosis.

Background: Endocardial lymphocytic infiltrates, known as Quilty effect, are a common finding of uncertain pathogenesis in cardiac allografts. Quilty effect was not observed before the use of cyclosporine A for immunosuppression and is not generally regarded as a manifestation of rejection. We hypothesized that the endocardial localization of Quilty effect may be related to a relative absence of cyclosporine A in this region., Methods: We used an indirect immunofluorescence staining method with rabbit polyclonal anti-cyclosporine A antibodies to detect cyclosporine A in fresh frozen sections of 27 cardiac allograft endomyocardial biopsies. Staining was graded 0 to +3. Negative controls were from untreated transplant candidates and from biopsies with the primary antibody omitted., Results: On comparison of endocardial and myocardial fluorescence in biopsy specimens from patients treated with cyclosporine A, there was less endocardial (0.7 +/- 1.1, p < 0.0001) than myocardial (2.2 +/- 0.5) staining. However, in biopsy specimens with Quilty effect (n = 12), this difference was significantly greater (endocardial = 0.2 +/- 0.6 versus myocardial = 2.3 +/- 0.5; p = 0.005) than in specimens without Quilty effect (n = 10) (endocardial = 1.4 +/- 1.2 versus myocardial = 2.1 +/- 0.6; p = 0.7). Endocardial thickness as measured by ocular micrometry was significantly greater in regions with (32 +/- 19 microns) than without (7 +/- 4 microns) Quilty effect, with involved regions showing increased connective tissue (p < 0.0001). In patients with and without Quilty effect, no differences in donor or recipient demographics, prevalence of diabetes, or plasma cyclosporine A levels were found., Conclusions: Although it has been postulated that Quilty effect is due to the presence of cyclosporine A in cardiac tissue (toxic effect or immunologic reaction), these data suggest that Quilty effect is related to reduced endocardial presence of cyclosporine A, leading to localized, contained, and usually not clinically significant endocardial rejection.

Within the first year, 15-20% of coronary artery saphenous bypass vein grafts (SVGs) occlude because of thrombosis or progressive intimal hyperplasia. One potential new strategy to reduce this complication would be to introduce antithrombotic or antiproliferative genes in vein grafts before implantation. The success of this approach requires an efficient DNA delivery system. In the present study we tested the feasibility of using adenovirus-transferrin/polylysine-DNA complexes (TfAdpl/DNA) to achieve high-efficiency gene transfer into vascular interposition vein grafts. All studies used the Escherichia coli LacZ (beta-galactosidase [beta-Gal]) reporter gene under the control of the cytomegalovirus (CMV) earlier promoter and enhancer (pCMV/LacZ). Autologous rabbit jugular vein segments were incubated ex vivo for 60 min in a solution of TfAdpl/DNA complexes (1.2 x 10(10) biotinylated adenovirus particles, 2,430 ng of streptavindylated polylysine. 10 micrograms of plasmid DNA, and 9 micrograms of transferrin-polylysine per ml), and then reimplanted across the ligated right carotid artery. Control veins were incubated in TfAdpl solution in which DNA was omitted. A total of six grafts were treated with TfAdpl/DNA, and two grafts were treated with TfAdpl. Veins were harvested 3 (n = 3) and 7 (n = 3) days later and beta-Gal activity was determined by X-Gal chromogen staining. All six TfAdpl/DNA-treated grafts stained intensely blue, whereas control grafts were negative. Microscopic examination of serial sections revealed intracellular blue granules consistent with beta-Gal activity to be present in all of the endothelial cells and in numerous medial and advential cells.(ABSTRACT TRUNCATED AT 250 WORDS)

We used monoclonal antibodies and immunohistochemical staining of frozen tissue sections to study the expression of cytokines in human cardiac allograft rejection. The 113 endomyocardial biopsy samples were stained for interleukin (IL)-2, IL-6, and interferon-gamma. The findings were compared to expression of the endothelial cell adhesion molecule ICAM-1, and the lymphocyte receptor for the adhesion molecule VCAM-1, VLA-4. Four biopsy samples from patients with idiopathic cardiomyopathy served as controls. IL-2 was not expressed in lymphocytes of controls and only occasionally in mild or moderate cellular rejection, humoral rejection, and Quilty lesions. IL-2 expression was prominent in severe cellular rejection. Interferon-gamma expression increased in proportion to the severity of cellular rejection and was not expressed in other conditions. IL-6 staining, which was only observed in occasional cases, was mild. Cytokine and adhesion molecule expression tended to increase with the severity of cellular rejection. This study shows that cytokine expression can be documented in human allograft endomyocardial biopsy samples with immunohistochemical techniques. The findings support the concept of an important role for cytokines in human cardiac allograft rejection.

Adhesion of leukocytes to vascular endothelial cells is a critical step in a variety of inflammatory conditions. We studied the expression and distribution of intercellular adhesion molecule-1 (ICAM-1) and endothelial leukocyte adhesion molecule-1 (ELAM-1) in frozen sections of 83 endomyocardial biopsy specimens from human allograft hearts using monoclonal antibodies and an avidin-biotin complex-alkaline phosphatase staining technique. Cases with cellular or humoral rejection and Quilty lesions were studied. Staining was graded from 0 to 3+ in lymphocytes and in capillary, arterial, venular, and endocardial endothelial cells. Expression of ICAM-1 in capillaries increased with the severity of cellular rejection and was prominent in humoral rejection. ICAM-1 was also expressed in lymphocytes in proportion to the degree of rejection. Little or no ELAM-1 expression was noted. In Quilty lesions the intensity of ICAM-1 expression was similar to that of mild-to-moderate rejection. Thus adhesion molecule expression can be identified in endomyocardial biopsy specimens of patients with rejection, suggesting a role for adhesion molecules in the process of rejection. These findings may prove useful in monitoring rejection and its response to therapy and in developing specific antisera directed against these molecules.