Michael acceptor molecules in natural products and their mechanism of action.

Purpose: Michael receptor molecules derived from plants are biologically active due to electrophilic groups in their structure. They can target nucleophilic residues on disease-related proteins, with significant therapeutic effects and low toxicity for many diseases. They provide a good option for relevant disease treatment. The aim of this study is to summarize the existing MAMs and their applications, and lay a foundation for the application of Michael receptor molecules in life science in the future. Methods: This review summarizes the published studies on Michael receptor molecules isolated from plants in literature databases such as CNKI, Wanfang Data, PubMed, Web of Science, ScienceDirect, and Wiley. Latin names of plants were verified through https://www.iplant.cn/. All relevant compound structures were verified through PubChem and literature, and illustrated with ChemDraw 20.0. Result: A total of 50 Michael receptor molecules derived from various plants were discussed. It was found that these compounds have similar pharmacological potential, most of them play a role through the Keap1-Nrf2-ARE pathway and the NF-κB pathway, and have biological activities such as antioxidant and anti-inflammatory. They can be used to treat inflammatory diseases and tumors. Conclusion: The Michael receptor molecule has electrophilicity due to its unsaturated aldehyde ketone structure, which can combine with nucleophilic residues on the protein to form complexes and activate or inhibit the protein pathway to play a physiological role. Michael receptor molecules can regulate the Keap1-Nrf2-ARE pathway and the NF-κB pathway. Michael receptor molecules can be used to treat diseases such as inflammation, cancer, oxidative stress, etc.

Mechanism of hyperproteinemia-induced blood cell homeostasis imbalance in an animal model.

Hyperproteinemia is a metabolic disorder associated with increased plasma protein concentration (PPC) and is often clinically complicated by malignant diseases or severe infections. At present, however, research on the molecular mechanism underlying high PPC (HPPC) is scant. Here, an animal model of primary hyperproteinemia was constructed in an invertebrate ( Bombyx mori) to investigate the effects of HPPC on circulating blood cells. Results showed that HPPC affected blood cell homeostasis, leading to increased reactive oxygen species levels, and induced programmed cell death dependent on the endoplasmic reticulum-calcium ion signaling pathway. HPPC induced the proliferation of blood cells, mainly granulocytes, by activating the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway. Supplementation with the endocrine hormone active substance 20E significantly reduced the impact of HPPC on blood cell homeostasis. Thus, we identified a novel signaling pathway by which HPPC affects blood cell homeostasis, which differs from hyperglycemia, hyperlipidemia, and hypercholesterolemia. In addition, we showed that down-regulation of gene expression of the hematopoietic factor Gcm could be used as a potential early detection indicator for hyperproteinemia.

Aberrant interhemispheric functional and structural connectivity in heroin-dependent individuals.

Models of heroin addiction emphasize the role of disrupted frontostriatal circuitry supporting cognitive control processes. However, heroin addiction-related alterations in functional and structural interactions among brain regions, especially between the cerebral hemispheres, are rarely examined directly. Resting-state functional magnetic resonance imaging (fMRI) approaches, which reveal patterns of coherent spontaneous fluctuations in the fMRI signal, offer a means to quantify directly functional interactions between the hemispheres. The corpus callosum (CC), which connects homologous regions of the cortex, is the major conduit for information transfer between the cerebral hemispheres and represents a structural connectivity index between hemispheres. We compared interhemispheric voxel-mirrored homotopic connectivity (VMHC) and CC volume between 45 heroin dependent-individuals (HDIs) and 35 non-addict individuals. We observed significant reduction of VMHC in a number of regions, particularly the striatum/limbic system regions, and significant decrease in splenium and genu sub-regions of CC in HDI. Importantly, within HDI, VMHC in the dorsal lateral prefrontal cortex (DLPFC) correlated with genu CC volume, VMHC in the putamen, VMHC in the DLPFC and genu CC volume and splenium CC volume were negatively correlated with heroin duration and impulsivity traits. Further analyses demonstrated that impairment of VMHC of bilateral DLPFC partially mediated the association between genu CC volumes decreased and increased impulsivity in HDI. Our results reveal a substantial impairment of interhemispheric coordination in the HDI. Further, interhemispheric connectivity correlated with the duration of heroin abuse and higher impulsivity behavior in HDI. Our findings provide insight into a heroin addicts' related pathophysiology and reinforce an integrative view of the interhemispheric cerebral functional and structural organization.

Larger corpus callosum and reduced orbitofrontal cortex homotopic connectivity in codeine cough syrup-dependent male adolescents and young adults.

OBJECTIVES: To characterize interhemispheric functional and anatomical connectivity and their relationships with impulsive behaviour in codeine-containing cough syrup (CCS)-dependent male adolescents and young adults. METHODS: We compared volumes of corpus callosum (CC) and its five subregion and voxel-mirrored homotopic functional connectivity (VMHC) in 33 CCS-dependent male adolescents and young adults and 38 healthy controls, group-matched for age, education and smoking status. Barratt impulsiveness scale (BIS.11) was used to assess participant impulsive behaviour. Abnormal CC subregions and VMHC revealed by group comparison were extracted and correlated with impulsive behaviour and duration of CCS use. RESULTS: We found selective increased mid-posterior CC volume in CCS-dependent male adolescents and young adults and detected decreased homotopic interhemispheric functional connectivity of medial orbitofrontal cortex (OFC). Moreover, impairment of VMHC was associated with the impulsive behaviour and correlated with the duration of CCS abuse in CCS-dependent male adolescents and young adults. CONCLUSIONS: These findings reveal CC abnormalities and disruption of interhemispheric homotopic connectivity in CCS-dependent male adolescents and young adults, which provide a novel insight into the impact of interhemispheric disconnectivity on impulsive behaviour in substance addiction pathophysiology. KEY POINTS: • CCS-dependent individuals (patients) had selective increased volumes of mid-posterior corpus callosum • Patients had attenuated interhemispheric homotopic FC (VMHC) of bilateral orbitofrontal cortex • Impairment of VMHC correlated with impulsive behaviour in patients • Impairment of VMHC correlated with the CCS duration in patients.

The impulsivity behavior is correlated with prefrontal cortex gray matter volume reduction in heroin-dependent individuals.

PURPOSE: Chronic exposure to heroin induced cerebral structural abnormalities may underlie heroin-related behaviors. The aim of this study was to: (1) identify cerebral structural abnormalities in heroin-dependent individuals (HDIs) by an automated and unbiased morphometric technique. (2) Define the correlation between these cerebral structural abnormalities and the impulsivity characteristic in HDIs. METHODS: 24 HDIs and 24 control subjects were completed with (1) high resolution structural magnetic resonance imaging scanning and analysis of gray matter volume using voxel-based morphometry implemented in Statistical Parametric Mapping and (2) a Chinese translation Barratt Impulsiveness Scale-11 questionnaire survey. Differences in regional gray matter volume were tested using an analysis of covariance model, co-varying for global gray matter and age. Statistical maps were set at p<0.05, corrected for multiple comparisons. The abnormal brain regions were correlated with the duration of heroin use and impulsivity scores. RESULTS: After adjusting for effects of age and total gray matter volume, cortical gray matter volume in the bilateral medial prefrontal cortex, bilateral dorsal lateral prefrontal cortex, and right fusiform cortex were significantly reduced in HDIs. Moreover, the gray matter volume in prefrontal cortex that showed group differences was negatively correlated with the duration of heroin use and negatively correlated with the impulsivity characteristic in HDIs. CONCLUSION: These findings reveal the prefrontal cortex was impaired in HDIs, meanwhile, indicate the changes in gray matter volume are relating to the duration of heroin use and the impulsivity characteristic of the HDIs.

Primary haemangiopericytoma outside muscle cone in fossa orbitalis: a case report and review.

PURPOSE: Orbital haemangiopericytoma (HPC) is a rare tumor with great histological variability and unpredictable clinical and biological behavior. The precise cell type origin is uncertain. METHOD: A case report is provided. RESULTS: A 24-year-old male patient visited an ophthalmologist because of proptosis, strabismus, and movement limitation of the right eye one year ago. These symptoms worsened rapidly. Histopathology, MRI methods and microscopy coil were applied to study the features of HPC. The tumor was removed and the histological examination revealed that it was a primary HPC of the orbit. CONCLUSION: MRI determined the size, location, circumscription, morphological detail, growth pattern, and relationship between masses and surrounding tissues. The genesis location of this HPC was rare, and its internal morphological characters were different from cases reported in the literature. The new type of microscope showed more details of tumor MR image characters objectively.

TRB3 gene silencing alleviates diabetic cardiomyopathy in a type 2 diabetic rat model.

OBJECTIVE: Tribbles 3 (TRB3) is associated with insulin resistance, an important trigger in the development of diabetic cardiomyopathy (DCM). We sought to determine whether TRB3 plays a major role in modulating DCM and the mechanisms involved. RESEARCH DESIGN AND METHODS: The type 2 diabetic rat model was induced by high-fat diet and low-dose streptozotocin. We evaluated the characteristics of type 2 DCM by serial echocardiography and metabolite tests, Western blot analysis for TRB3 expression, and histopathologic analyses of cardiomyocyte density, lipids accumulation, cardiac inflammation, and fibrosis area. We then used gene silencing to investigate the role of TRB3 in the pathophysiologic features of DCM. RESULTS: Rats with DCM showed severe insulin resistance, left ventricular dysfunction, aberrant lipids deposition, cardiac inflammation, fibrosis, and TRB3 overexpression. We found that the silencing of TRB3 ameliorated metabolic disturbance and insulin resistance; myocardial hypertrophy, lipids accumulation, inflammation, fibrosis, and elevated collagen I-to-III content ratio in DCM rats were significantly decreased. These anatomic findings were accompanied by significant improvements in cardiac function. Furthermore, with TRB3 gene silencing, the inhibited phosphorylation of Akt was restored and the increased phosphorylation of extracellular signal-regulated kinase 1/2 and Jun NH(2)-terminal kinase in DCM was significantly decreased. CONCLUSIONS: TRB3 gene silencing may exert a protective effect on DCM by improving selective insulin resistance, implicating its potential role for treatment of human DCM.

Regional homogeneity changes in heroin-dependent individuals: resting-state functional MR imaging study.

PURPOSE: To identify heroin-related modulations of neural activity in the resting state in heroin-dependent individuals (HDIs) by using resting-state functional magnetic resonance (MR) imaging and a regional homogeneity method and to investigate whether these changes of neural activity can be related to duration of heroin use and to decision-making deficits in HDIs. MATERIALS AND METHODS: This prospective study was approved by the appropriate ethics committee, and written informed consent was obtained from each participant. Thirty-one HDIs receiving methadone-maintained treatment and 24 control subjects participated. Resting-state functional MR imaging was performed by using a gradient-echo echo-planar imaging sequence. Regional homogeneity was calculated by using software. Voxel-based analysis of the regional homogeneity maps between control and HDI groups was performed with two-sample t tests by using software. Statistical maps were set at P less than .05 and were corrected for multiple comparisons. The Iowa gambling task (IGT) was used to assess participant decision making during uncertainty. Abnormal clusters revealed by group comparison were extracted and correlated with behavioral performance at the IGT and with duration of heroin use. RESULTS: Regional homogeneity was diminished in the bilateral medial orbitofrontal cortex (OFC), bilateral dorsal medial thalamus, bilateral cuneus, and lingual gyrus in HDIs compared with control subjects. There were negative correlations between mean regional homogeneity in the medial OFC, bilateral cuneus, and lingual gyrus and duration of heroin use. There was a positive correlation between mean regional homogeneity in the medial OFC and performance level at the IGT. CONCLUSION: The present study reveals resting-state abnormalities in HDIs that may lead to further improvement of the understanding of the neural substrates of cognitive impairment in HDIs.

Amplitude low-frequency oscillation abnormalities in the heroin users: a resting state fMRI study.

Functional neuroimaging studies have revealed abnormal functional organization of the heroin users' brain, including reward circuit, cognitive control circuit, memory circuit, motivation and salience evaluation circuits and so on. In the current study, we aimed to explore the functional changes in the regional brain of heroin users using the amplitude of low-frequency oscillations in the Blood Oxygenation Level Dependent (BOLD) signals. With fMRI data acquired during resting state from 24 chronic heroin users (all subjects were being treated with methadone) and 24 non-addicted controls, we investigated addiction related altered in the amplitude low-frequency fluctuate (ALFF) between the two groups. Compared with controls, we found that heroin addicts had decreased ALFF in the bilateral dorsal anterior cingulate cortex (dACC), bilateral medial orbit frontal cortex (mOFC), left dorsal lateral prefrontal cortex (dlPFC), left middle temporal gyrus, left inferior temporal gyrus, posterior cingulate cortex and left cuneus as well as increased ALFF in the bilateral angular gyrus, bilateral precuneus, bilateral supramarginal gyrus, left post cingulate cortex and left middle frontal gyrus. Moreover, we also found that the increased ALFF in the bilateral parietal lobe had a significantly positive correlation with the methadone does, thus we inferred that the reduced ALFF may due to heroin consumption, nevertheless, the increased ALFF in the bilateral parietal lobe may have resulted from the methadone treatment. This resting-state fMRI study suggests that the changed spontaneous neuronal activity of these regions may be implicated in the underlying pathophysiology of heroin addicts.

[Correlation analysis between multi-slice CT perfusion imaging and microvessel density in ovarian tumors].

OBJECTIVE: To analyze the correlation between the perfusion data and microvessel density (MVD) in ovarian tumors, and investigate the hemodynamic features of the tumors in terms of anatomy and functional CT imaging. METHODS: Six patients with surgically confirmed benign ovarian tumors and 6 with malignant ovarian tumors underwent multi-slice CT perfusion imaging to acquire the perfusion parameters including perfusion, PEI, TTP, BV peak enhancement image(PEI), time to peak(TTP) and blood volume(BV). The tumors were stained and counted by Immunohistochemical staining of the microvessels in the tumor was performed to detect the MVD. RESULTS: s The time-density curves of the benign ovarian tumors increased slowly, reaching the peak at 40 s; the curves of the malignant tumors rose rapidly and continuously and reached the peak at 25 s. The differences in the perfusion data (PEI, TTP, BV) were statistically significant between the benign and malignant tumors (P<0.05). The MVD of the malignant tumors was significantly greater than that of the benign tumors (P<0.05). The mean BV of the malignant ovarian tumor was positively correlated to MVD (r=0.786, P<0.05). CONCLUSION: Multi-slice spiral CT perfusion imaging can provide accurate enhancement data of the ovarian tumors and helps in the diagnosis and differential diagnosis of the ovarian tumors by presenting the changes of the hemodynamic features in the tumors.

[Atypical magnetic resonance imaging vs pathological findings of leiomyoma in the female reproductive system].

OBJECTIVE: To investigate magnetic resonance imaging (MRI) findings of the atypical leiomyoma in the female reproductive system in comparison with the pathological features of the neoplasms. METHODS: A retrospective analysis of the MRI findings and the pathological features was conducted in 24 cases of atypical leiomyoma involving the female reproductive system. RESULTS: Atypical leiomyomas were displayed by MRI as solid tumor mass surrounded by cystic degeneration, pseudotumors, or solid mass with homogeneous signal intensity. Intrauterine lesions were found in 19 cases, involving the subserosal layer (n=11), intramural region (n=4), broad ligament (n=3), cervix (n=2), submucous layer (n=2), vagina (n=1), and the ovary (n=1). Except for two cases with submucous lesions shown as solid mass, all the cases had lesions appearing as solid cystic mass, whose solid part showed hypo or isointense signals on T1WI and moderate hyperintense signals on T2WI, with heterogeneous enhancement after contrast agent injection. Tumor cell and interstitial cell swelling, vascular hyalinosis, hyalinosis, myxoedema, cystic degeneration, and hemorrhage were found in the lesions. CONCLUSION: Leiomyoma can occur at almost any site in the female reproductive system, and atypical leiomyoma usually are shown as solid cystic mixed mass in the pelvic cavity. Evaluation of the relationship between the solid mass and cystic portion and observation for the presence of low signal on T2WI may help in the diagnosis of atypical leiomyoma.

Effect of atorvastatin versus rosuvastatin on levels of serum lipids, inflammatory markers and adiponectin in patients with hypercholesterolemia.

PURPOSE: To compare the short-term effect of treatment with atorvastatin and rosuvastatin on levels of serum lipids, inflammatory markers and adiponectin in patients with hypercholesterolemia. METHODS: Sixty-nine patients with hypercholesterolemia were randomly assigned to receive 10 mg/day of atorvastatin or rosuvastatin for 12 weeks. Inflammatory biomarkers, including highsensitivity C-reactive protein (hs-CRP), tumor necrosis factor (TNF)-alpha, matrix metalloproteinase-9 (MMP-9), and endothelin (ET-1), plasminogen activator inhibitor type 1 (PAI-1) and plasma tissue plasminogen activator (tPA), adiponectin, and lipid profiles were measured before and after statin therapy. RESULTS: Atorvastatin and rosuvastatin both lowered levels of hs-CRP, MMP-9, PAI-1, total cholesterol (TC), and low-density lipoprotein cholesterol (LDL-C) from baseline values, with rosuvastatin lowering TC and LDL-C to a greater extent than atorvastatin (P < 0.05). Adiponectin level increase was 15% higher than that at baseline with atorvastatin (P > 0.05) but 67% higher with rosuvastatin (P < 0.05). CONCLUSIONS: Therapy with both statins not only significantly improved lipid profiles but also decreased levels of vascular biomarkers hs-CRP, MMP-9, and PAI-1; however, only rosuvastatin increased serum adiponectin levels significantly in patients with hypercholesterolemia, which could imply a beneficial effect in coronary artery disease.

[Anatomic study of the facial artery using multislice spiral CT angiography].

OBJECTIVE: To study the anatomy of the facial artery using 16-slice spiral CT angiography (CTA). METHODS: Forty-five patients without vascular pathologies or other anomalies underwent 16-slice spiral CT angiography after injection of 350 mg I/ml Omnipaque through the ulnar vein. MIP and volume rendering of the images were performed to analyze the left and right facial artery. RESULTS: The left facial artery arose from the external carotid artery with the distances from the carotid artery bifurcation of 4.5-47.90 mm (mean 18.77-/+8.98 mm), and in 1 case (2.2%), the artery arose from the common carotid artery. The right facial artery arose from the external carotid artery in all the 45 cases (100%) with distances from the carotid artery bifurcation of 6.8-39.70 mm (mean 19.23-/+8.25 mm). The bilateral facial arteries more commonly arose from the external carotid artery independently, and the lingual artery and facial artery sharing the same trunk arising from the external carotid artery was less common. In 1 case, the left facial artery and the submental artery shared the same trunk, and in another case, the left facial artery, thyroid artery and lingual artery shared the same trunk. The diameter of the left facial artery ranged from 1.40 to 4.70 mm (mean 2.83-/+0.77 mm), and that of the right facial artery was 1.60-4.30 mm (mean 2.81-/+0.79 mm). The left facial artery ended below the angle of the mouth in 12 cases (26.67%), between the angle of the mouth and the nasal wing in 7 cases (15.56%), and above the nasal wing in 26 cases (57.77%). The right facial artery ended below the angle of the mouth in 7 cases (15.56%), between the angle of the mouth and the nasal wing in 12 cases (26.67%), and above the nasal wing in 26 cases (57.77%). The bilateral facial arteries frequently passed through the submandibular gland parenchyma (23 cases on the left and 24 cases on the right), and the facial arteries were found occasionally to run below the submandibular gland (11 cases on the left and 9 on the right). CONCLUSION: 16-slice spiral CT angiography can help in preoperative facial artery evaluation noninvasively. This modality can clearly display the bilateral facial arteries, including their origin, course, distribution and ending to provide detailed information for flap designing before plastic surgery and for preoperative evaluation for microvascular surgery.

[Influences of bcr-abl gene vaccine on inoculated SP2/0/bcr-abl tumor cells in mice].

To study the influence of vaccine of bcr-abl fusion gene fragment on inoculated SP2/0/bcr-abl tumor cells in mice, BALB/c mice were immunized with pVbcr-abl, pVbcr-abl/mIL7 plasmids, respectively, then SP2/0/bcr-abl cells expressing the fragment of bcr-abl fusion gene were inoculated subcutaneously into the groin of BALB/c mice in order to observe the effect of vaccine on growth of inoculated SP2/0/bcr-abl tumor cells. The results showed that there were distinct differences on the time of tumor growth, the time of tumor ulceration, tumor volume and survival time of mice bearing tumor between two immunized groups and two control groups (blank and vacant plasmid groups). The mice immunized with pVbcr-abl/mIL7 lived longer as compared to mice immunized with pVbcr-abl. The tissue of inoculated tumor was more compact, tumor organ was larger, tumor form was irregular in 2 control groups, while the tissue of inoculated tumor was looser, tumor volume was smaller, and with mass inflammatory infiltration in two immunized groups. Moreover, the metastatic tumor cells were found in the livers of control groups, but not observed in two immunized groups. It is concluded that the protection occurred in immunized mice which inhibited the growth of SP2/0/bcr-abl tumor cell in vivo.

[PPARgamma1 overexpression on caveolin-1 expression of Raw264.7 cells].

OBJECTIVE: To investigate the effect of PPARgamma1 gene overexpression on caveolin-1 mRNA and protein expressions in a murine macrophage cell line Raw264.7. METHODS: Replication-deficient recombinant adenovirus expression vector of PPARgamma1 was constructed using the AdEasy system. Raw264.7 cells were randomly treated as follows: P group (PPARgamma1 gene overexpression), T group (Troglitazone 40 micromol/L in DMSO), PT group (PPARgamma1 gene overexpression and Troglitazone) and control group. Changes of PPARgamma1 and caveolin-1 at mRNA and protein levels were investigated. RESULTS: Caveolin-1 expression can be detected by RT-PCR in Raw264.7, by immunocytochemistry method in cell and nuclear membrane but not by immunoblotting at protein level. Caveolin-1 expression at mRNA and protein levels in Raw264.7 were significantly higher in P, T and PT groups compared to control group and the expression was also significantly higher in PT group than that in P group and T group (P < 0.05). PPARgamma expression was significantly increased in PT group and P group where remained unchanged in T group compared to control group. CONCLUSION: PPARgamma1 overexpression can upregulate caveolin-1 expression in macrophages. Troglitazone upregulated caveolin-1 expression in the absence of increased PPARgamma1 expressions at mRNA and protein levels.

[Recombinant eukaryotic expression plasmid of bcr-abl gene fragment induces specific immune response in mice].

OBJECTIVE: To study the specific immune response induced by a recombinant eukaryotic expression plasmid encoding bcr-abl fusion gene fragment so as to explore new immunotherapy in mouse. METHODS: A recombinant eukaryotic vector pVbcr-abl expression cDNA fragment of bcr-abl fusion gene was constructed and used to immunize BALB/c mice. Serum level of bcr-abl specific antibody was detected by enzyme-linked immunosorbent assay (ELISA). Twenty days later the immunized mice were subcutaneously inoculated SP2/0/bcr-abl cells. The survival time, tumor growth time and lymphocytic infiltration were observed. T cells infiltration into tumor tissue was analyzed by immunohistochemistry. Changes of T cell subset in the spleen of mice was analyzed by fluorescent-activated cell sorting (FACS) and the cytotoxicity T lymphocyte (CTL) activity in spleen by lactate dehydrogenase (LDH)-release assay. RESULTS: The eukaryotic expression vector pVbcr-abl was constructed successfully, and highly expressed the cDNA fragment of bcr-abl fusion gene. The BALB/c mice immunized with the vector could generate the specific antibody and CTL, resulting in a specific immunoprotection. There were dramatic differences in the tumor-forming time, tumor ulcer appearing time and tumor-growing speed between the immunized and the control groups. The mice had longer survival time in the immunized group than in the control group. There were a large amount of CD3(+) T cells infiltration in tumor tissue of the immunized mice. The spleen cells from the immunized mice had higher CTL activity with a alteration of T cell subset, the CD4(+)/CD8(+) ratio being 1.54 +/- 0.29, higher than that of control group (1.18 +/- 0.30). CONCLUSION: The recombinant eukaryotic expression plasmid pVbcr-abl can induce in vivo not only the generation of specific antibody, but also high level of specific CTL activity, resulting in killing the SP2/0/bcr-abl tumor cells directly and inhibiting the tumor growth.

[Genetic transformation of peach immature cotyledons with its antisenes ACO gene].

Genetic transformation of peach immature cotyledons with its ACO antisense gene was studied by using particle bombardment method through Agrobacterium tumefaciens. Kanr shoots and Kanr plantlet were obtained. The plantet with ACO antisenes gene through Agrobacterium tumefaciens was obtained by micrografting technique and survived for nearly one month. The results of the PCR, PCR-southern, genomes southern hybridization analysis and GUS color reaction of some Kanr materials showed in some degree that the peach ACO antisens gene was integrated into peach genomes.