Molecular cloning and characterization of fruit specific promoter from <i>Cucumis sativus</i>L.

The isolation and characterization of fruit specific promoters are critical for the manipulation of nutritional value and agronomic quality of fruits by genetic engineering and also opened a new era in edible vaccine technology. Expansins are proteins that induce loosening of individual plant cells by disrupting the non-covalent interactions between cellulose and hemicellulose microfibrils and hence have role in growth programs including fruit ripening. We report the identification of an expansin gene (CsExp) from Cucumis sativus that exhibits high levels of mRNA abundance and is specifically expressed in ripened fruit. The promoter region of CsExp also contains elements responsible for its fruit specific expression. Transient expression studies of the CsExp promoter were conducted with particle bombardment, followed by GUS histochemical assay and real time PCR. CaMV35S promoter was used as the positive control in all these experiments. Clear fruit specificity was observed for CsExp promoter in all the experiments. Thus CsExp promoter from Cucumber is a good candidate to target expression of the foreign genes to engineer fruit specific traits.

Interplay between mesenchymal stem cells and macrophages:Promoting bone tissue repair

The repair of bone tissue damage is a complex process that is well-orchestrated in time and space,a focus and difficulty in orthopedic treatment.In recent years,the success of mesenchymal stem cells(MSCs)-mediated bone repair in clinical trials of large-area bone defects and bone necrosis has made it a candidate in bone tissue repair engineering and regenerative medicine.MSCs are closely related to macrophages.On one hand,MSCs regulate the immune regulatory function by influencing macrophages proliferation,infiltration,and phenotype polarization,while also affecting the osteoclasts differentiation of macrophages.On the other hand,macrophages activate MSCs and mediate the multilineage differentiation of MSCs by regulating the immune microenvironment.The cross-talk between MSCs and macrophages plays a crucial role in regulating the immune system and in promoting tissue regeneration.Making full use of the relationship between MSCs and macrophages will enhance the efficacy of MSCs therapy in bone tissue repair,and will also provide a reference for further application of MSCs in other diseases.

Syzygium cumini(L.)Skeels:A review of its phytochemical constituents and traditional uses

Syzygium cumini(S.cumini)(L.) Skeels(jambolan) is one of the widely used medicinal plants in the treatment of various diseases in particular diabetes.The present review has been primed to describe the existing data on the information on botany,phytochemical constituents,traditional uses and pharmacological actions of 5.cumini(L.) Skeels(jambolan).Electronic database search was conducted with the search terms of Eugenia jambolana,S.cumini,jambolan,common plum and java plum.The plant has been viewed as an antidiabetic plant since it became commercially available several decades ago.During last four decades,numerous folk medicine and scientific reports on the antidiabetic effects of this plant have been cited in the literature.The plant is rich in compounds containing anthocyanins,glucoside,ellagic acid,isoquercetin,kaemferol and myrecetin.The seeds are claimed to contain alkaloid,jambosine,and glycoside jambolin or antimellin,which halts the diastalic conversion of starch into sugar.The vast number of literatures found in the database revealed that the extracts of different parts of jambolan showed significant pharmacological actions.We suggest that there is a need for further investigation to isolate active principles which confer the pharmacological action.Hence identification of such active compounds is useful for producing safer drugs in the treatment of various ailments including diabetes.

Chromium detoxification mechanism induced growth and antioxidant responses in vetiver(Chrysopogon zizanioides(L.) Roberty)

This study investigated the chromium(Cr)detoxification mechanism-induced changes in growth and antioxidant defence enzyme activities in Chrysopogon zizanioides.Plant growth decreased by 36.8%and 45.0%in the shoots and roots,respectively,in the 50 mg/L Cr treatment.Cr accumulation was higher in root(9807μg/g DW)than in shoots(8730μg/g DW).Photosynthetic pigments and malondialdehyde content increased up to the 30 mg/L Cr treatment,whereas they declined at higher doses.The activity of superoxide dismutase(SOD),catalase(CAT)and peroxidase(POX)were increased significantly with increasing of Cr dose but slightly declined at higher doses.Isozyme banding patterns revealed the expression of multiple bands for SOD,CAT and POX enzymes,and the band intensity decreased at high doses of Cr exposure.These results indicate that higher Cr doses increased the oxidative stress by over production of reactive oxygen species(ROS)in vetiver that had potential tolerance mechanism to Cr as evidenced by enhanced level of antioxidative enzymes,photosynthetic pigments,MDA contents.Therefore,vetiver has evolved a mechanism for detoxification and accumulates higher concentration of toxic Cr.This study provides a better understanding of how vetiver detoxifies Cr.

Detection and clinical significance of multidrug resistance-1 mRNA in bone marrow cells in children with acute lymphoblastic leukemia by real-time fluorescence quantitative RT-PCR

Objective： Multidrug resistance（MDR） is one of the most important reasons for treatment failure and recurrence of acute leukemia. Its manifestations are different in children with acute lymphoblastic leukemia（ALL） which may be due to different detection methods. This study was to detect the expression of MDR1 mRNA in bone marrow cells of children with ALL by real-time fluorescence- quantitative reverse transcription polymerase-chain reaction（FQ-RT-PCR）, and combine minimal residual desease（MRD） detection by flow cytometry（FCM） and to study their relationship with treatment response and prognosis of ALL. Methods：The MDR1 mRNA levels in bone marrow cells from 67 children with ALL[28 had newly diagnosed disease, 27 had achieved complete remission（CR）, 12 recurrent] and 22 children without leukemia were detected by FQ-RT-PCR. MRD was detected by FCM. The patients were observed for 9-101 months, with a median of 64 months. Results：Standard curves of human MDR1 and GAPDH genes were constructed successfully. MDR1 mRNA was detected in all children with a positive rate of 100%. The mRNA level of MDR1 was similar among the newly diagnosed ALL group, CR group, and control group（P 〉 0.05）, but significantly higher in the recurrence group than that in newly diagnosed disease group and control group（0.50 ± 0.55 vs. 0.09 ± 0.26 and 0.12 ± 0.23, P〈 0.05）. 54 ALL patients were followed up, and it was found that MDR1 mRNA level was significantly higher in ALL patients within 3 years duration than that of ALL patients with 3-6 years and over 6 years duration（0.63 ± 0.56 vs. 0.11 ± 0.12 and 0.04 ± 0.06, P〈 0.01）. For the 28 children with newly diagnosed disease, the MDR1 mRNA level was similar between WBC 〉 50 ~ 109 group and WBC〈50 × 10^9 group（P〉 0.05）. In the 33 CR patients, the MDR1 mRNA level was significantly higher in MRD〉10a group than that in MRD〈10a group（0.39 ± 0.47 vs. 0.03 ± 0.03, P 〈 0.05）. Conclusion：The sensitivity and specificity of FQ-RT-PCR in detecting MDR1 mRNA in bone marrowy cells of children with ALL patients are high. MDR1 mRNA is expressed in children with and without leukemia. MDR1 mRNA is highly expressed in the CR ALL patients with high MRD, recurrence and short duration（within 3 years）. Monitoring MRD and the MDR1 mRNA level might be helpful for individual treatment.

Diagnostic value of a panel of tumor markers as a part of a diagnostic work-up for ascites of unknown etiology

Background & objectives: Data regarding tumor marker usefulness in diagnosing ascites of unknown etiology and determining its malignant nature are conflicting. We aim to assess the diagnostic value of ascitic and serum tumor markers in ascites of unknown etiology and to evaluate their usefulness besides other laboratory tests in a diagnostic work-up in those patients. Design & setting: A prospective case-control study conducted at Assiut University hospital and oncology institute. Patients & Methods: Three groups were included;Group I: 41 patients with ascites of unknown etiology Group II: 7 Patients with TB ascites and Group III:14 patients with cirrhotic ascites. We assessed the CEA, CEA mRNA, CA15-3, CA19.9, CA125, AFP and PSA in serum and ascetic fluid. A diagnostic work-up for group I included: IPD test, ultrasound, CT, ascetic fluid cytology, SAAG, Laparotomy and biopsy. Results: Ascetic fluid and serum levels of CA15-3 and CA125 were significantly increased in group I and were significantly increased in histopathologically proved malignant ascites compared to TB and cirrhotic ascites. In group I, CA 125 was significantly higher in ascites than serum. With the exception of PSA, all tumor markers significantly correlated in serum and ascetic fluid. No significant difference in the level of ascetic CEA messenger RNA was detected between the 3 groups. Cytology had 53% sensitivity, 94% specificity and CA 125 & CA15-3 had 81% sensitivity and 75% specificity in detection of malignant ascites repectively. Laparotomy and biopsy: diagnosed malignnancy in 53.3% and TB in 13.3%. Conclusions: A diagnostic work-up including SAAG, tumor markers in the serum and ascetic fluid may help in adjunct with ascetic fluid cytology, laparotomy and biopsy, imag- ing and other laboratory tests in diagnosing ascites of unknown etiology.

MINOCYCLINE HAS A NEUROPROTEC- TIVE EFFECT ON RETINAL GANGLION CELLS SURVIVAL AFTER OPTIC NERVE TRANSECTION IN RATS

Purpose: To evaluate the neuroprotective effect of Minocycline on retinal ganglion cells (RGCs) survival after optic nerve tran-section in rats. Minocycline is commonly used by humans, because of its beneficial antimicrobial and anti-inflammatory actions. Recently, it was shown that it has remarkable neuroprotective qualities in animal models of cerebral ischemia,

Acute and Chronic Toxicity of Thioacteamide and Alterations in Blood Cell Indices in Rats

Background: Thioacetamide (TAA) has been used extensively in the development of suitable animal models of acute and chronic liver injury employing various doses, times and routes of its administration, particularly in drinking water due to its resemblance with human liver fibrosis and cirrhosis. The aim of this study was to investigate and compare hematological alteration during the acute and chronic liver inflammation. Methods: Acute Liver inflammation was induced in Wistar rats via intraperitoneal injection of thioacetamide and the animals were sacrificed 12 h after the TAA administration. Induction of chronic liver inflammation was performed by continuous administration of TAA in the drinking water (200 mg/L) during 18 weeks of experiment. After that all animals were sacrificed and Blood samples were collected for further analysis. Results: Single intra peritoneal injection of TAA (300 mg/kg B.W.) induced an acute condition with hematological changes including leukocytosis with marked neutrophilia (P = 0.0429), lymphopenia, thrombocytosis as well as increased hemoglobin concentration (P 0.05) and decline of erythrocytic count (P = 0.0009). Eighteen weeks of uninterrupted supply of TAA (200 mg/L) in drinking water lead to chronic inflammation and the hematological alterations were leucopenia (P = 0.0197) accompanied with neutropenia and thrombocytopenia. Increase in RBCs (P = 0.0073) and Hb contents was also observed with a decline of red cell indices. Conclusion: Taken together these findings we can conclude that the animals respond differently under acute and chronic inflammatory condition with TAA administration. Leukocytosis with marked neutrophilia, thrombocytosis as well as increased hemoglobin concentration and decline of erythrocytic count were observed in acute while leucopenia accompanied with neutropenia and thrombocytopenia and increase in RBCs, Hb and Hct was also observed with a decline of other red cell indices during chronic phase.

Malignant Transformation of Human Embryonic Liver Cells Induced by Hepatitis B Virus and Aflatoxin B_1

In order to investigate the effect of hepatitis B virus (HBV) and aflatoxin B 1 (AFB 1) on hepatocarcinogenesis, the human embryonic liver cells infected with HBV were transplanted to nude mice by subcutaneous route and the transplanted mice were divided into 4 groups for study, in which the group A of mice was injected with HBV-infected human embryonic liver cells and followed by injections of AFB 1 once a week (HBV+AFB 1); the group B was treated with HBV as group A, but no AFB 1 was given (HBV +); the group C was injected with normal human embryonic liver cells and AFB 1 was used as group (AFB 1 +) and the group D or control group was injected with normal embryonic liver cells without addition of AFB 1. The experimental results showed that the incidences of tumor formation in different groups were 27.3% (6/22) in group A; 0% (0/13) in group B; 13.3% (2/15) in group C and 0% (0/14) in group D respectively. All the tumors formed were proved to be human hepatocellular carcinoma (HCC) by pathological examinations and the tumor tissues were anthrogenetic as demonstrated by EMA monoclonal antibody. The HBV-X and HBV-S genes could be detected in the tumor tissues by means of slot hybridization and PCR amplification, indicating that the HBV-DNA genes had integrated into DNA of host cells. Thus, we have successfully induced the human HCC through HBV infection and introduction of AFB 1 with a synergistic effect between HBV and AFB 1 in hepatocarcinogenesis.

p53 INDEPENDENT G_1 ARREST AND APOPTOSIS INDUCED BY ADRIAMYCIN

The biological activity of adriamycin was investigated in human breast carcinoma (HBC) cells. Adri-amycin inhibited the growth of a number of HBC cell lines and induced G1 arrest followed by apoptosis. In MCF-7 cells that harbor wild-type p53, adriamycin-induced G1 arrest and apoptosis was accompanied by p53-independent regulation of WAF1/CIP1 as well as bax mRNA levels. In MDA-MB-231 cells which possess a mutant p53, adriamycin-induced G1 arrest and apoptosis was also associated with a cpncomitant up-regulation of WAF1/CIP1 mRNA while these cells did not express bax or bcl-2 messages. Thus, adriamycin induces G1 arrest and apoptosis via a unique pathway which appears to involve activation of downstream effectors of p53-independent manner.

Rewilding staple crops for the lost halophytism:Toward sustainability and profitability of agricultural production systems

Abiotic stress tolerance has been weakened during the domestication of all major staple crops.Soil salinity is a major environmental constraint that impacts over half of the world population;however,given the increasing reliance on irrigation and the lack of available freshwater,agriculture in the 21st century will increasingly become saline.Therefore,global food security is critically dependent on the ability of plant breeders to create high-yielding staple crop varieties that will incorporate salinity tolerance traits and ac-count for future climate scenarios.Previously,we have argued that the current agricultural practices and reliance on crops that exclude salt from uptake is counterproductive and environmentally unsustainable,and thus called for a need for a major shift in a breeding paradigm to incorporate some halophytic traits that were present in wild relatives but were lost in modern crops during domestication.In this review,we provide a comprehensive physiological and molecular analysis of the key traits conferring crop halophy-tism,such as vacuolar Na+sequestration,ROS desensitization,succulence,metabolic photosynthetic switch,and salt deposition in trichomes,and discuss the strategies for incorporating them into elite germ-plasm,to address a pressing issue of boosting plant salinity tolerance.