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In order to improve the salt tolerance of banana NHX genes, we cloned a MaNHX5 gene from Musa acuminata L. AAA group and predicted the key salt-tolerant amino acid sites and mutant protein structure changes of MaNHX5 by using bioinformatics tools. The 276-position serine (S) of MaNHX5 protein was successfully mutated to aspartic acid (D) by site-directed mutagenesis, and the AXT3 salt-sensitive mutant yeast was used for a functional complementation test. The results showed that after the mutated MaNHX5 gene was transferred to AXT3 salt-sensitive mutant yeast, the salt tolerance of the mutant yeast was significantly improved under 200 mmol/L NaCl treatment. It is hypothesized that Ser276 of MaNHX5 protein plays an important role in the transport of Na+ across the tonoplast.
Subject(s)
Amino Acids/metabolism , Gene Expression Regulation, Plant , Musa/metabolism , Plant Proteins/metabolism , Plants, Genetically Modified , Saccharomyces cerevisiae/metabolismABSTRACT
Objective::To investigate the effect of Xiaoyaosan (XYS) on hepatic lipid metabolism and steatohepatitis in ovariectomized (OVX) female SD rats and its mechanism. Method::Forty female SD rats were randomly divided into sham surgery group, OVX group, low-dose XYS group (3 g·kg-1), and high-dose XYS group (9 g·kg-1). Bilateral ovaries of rats were excised to replicate the obesity model of ovariectomized rat. After 6 weeks of intragastric administration, the change rate of body mass in each group, the levels of blood lipids and liver function of rats were detected. Hematoxylin-eosin (HE) staining and oil red staining were used to observe the hepatocyte histomorphology and the intrahepatic fatty deposits. The expressions of hepatic proinflammatory cytokines and estrogen receptor beta (ERβ) were determined by quantitative real time polymerase chain reaction (Real-time PCR). Result::Compared with sham surgery group, the change rate of body mass of OVX group was significantly increased (2-6 weeks) with the changes in the course of drug administration and the levels of serum total cholesterol (TC), alanine aminotransferase (ALT) (P<0.05), aspartate amino transferase (AST) (P<0.01), low-density lipoprotein (LDL) (P<0.05) were markedly increased too (P<0.05, P<0.01). By histological method, in OVX group, the structure of hepatic cord became disordered, and there were new lipid droplets in hepatocyte cytoplasm, transcription levels of hepatic interleukin-1β (IL-1β) and interleukin-6 (IL-6) in OVX group were significantly increased (P<0.05). Compared with OVX group, the growth rate of body weight in low-dose and high-dose XYS group showed significant decreases with the increase of the cycle of drug administration (3-6 weeks). XYS significantly reduced levels of serum TC, ALT, AST, and LDL levels of OVX rats (P<0.05) in a dose-dependent manner, while serum triglyceride (TG), alkaline phosphatase (AKP) and high-density lipoprotein (HDL) levels in the four groups showed no statistical significance, XYS can improve hepatocyte structure and steatosis of OVX rats, XYS could reduce the transcription hepatic levels of IL-6 and IL-1β of OVX rats in a dose-dependent manner (P<0.05, P<0.01), but there was no significant difference in the transcription level of tumor necrosis factor-α (TNF-α) among groups, both low and high-dose XYS can increase the transcription hepatic level of ERβ in OVX group (P<0.05, P<0.01). Conclusion::XYS can improve the growth rate of body mass, the hepatic lipid metabolism abnormalities and steatohepatitis of OVX rats. The mechanism may be related to the elevated expression of hepatic ERβ by XYS, so as to inhibit the hepatic pro-inflammatory factors expressions.
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OBJECTIVE@#To evaluate the safety and efficacy of allogeneic natural killer (NK) cells in the treatment of primary hepatocellular carcinoma (HCC), and to elucidate the mechanism of NK cells therapy.@*METHODS@#Twenty-one patients with primary HCC treated with allogeneic NK cells at the Fifth Medical Center of the PLA General Hospital were followed up for 1 year. Peripheral blood mononuclear cells (PBMCs) were isolated from patient-related donors and cultured in vitro for 15 days and infused to the patients in two consecutive days. Clinical data and laboratory data were collected and analyzed, including survival, clinical features, imaging changes, hematology, immunology, and biochemical indicators to evaluate the safety and efficacy of allogeneic NK cell therapy. The changes of peripheral blood lymphocyte subsets after treatment were also analyzed to explore the possible anti-tumor mechanisms.@*RESULTS@#(1) Of the 21 patients with primary HCC, 11 patients were treated once, 5 patients were treated twice, and 5 patients were treated 3 times. After allogeneic NK cells infusion, 10 patients had fever, 1 patient had slight hepatalgia and 1 patient had slight headache, no other adverse events occurred including acute and chronic graft-versus-host disease (GVHD). They resolved spontaneously within 8 hours without other treatment. (2) The total disease control rate was 76.2% during one-year follow-up. Among them, the patients with Barcelona clinic liver cancer (BCLC) stage A had a disease control rate of 100%, stable disease (SD) in 10 cases; BCLC stage B patients had a disease control rate of 60%, partial response (PR) in 1 case, and SD 2 in cases; BCLC stage C patients had a disease control rate of 50%, complete response (CR) in 1 case, and 2 cases of PR. (3) The frequencies of NK cells and CD8+ T cells in peripheral blood were significantly lower than that before at 24 hours after treatment, and the frequencies of CD4+ T cells and CD4/CD8 were significantly higher than the baseline.@*CONCLUSION@#Allogeneic NK cells have good safety and efficacy in the treatment of primary HCC. The anti-tumor effect of the allogeneic NK cells may play an important role in the activation of the patient's natural immune system and delay disease progression, suggesting that allogeneic NK cells combined with sorafenib may be a very effective treatment for advanced HCC, and further large-sample multicenter randomized controlled clinical trials are needed to validate this result.
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Humans , Carcinoma, Hepatocellular , Graft vs Host Disease , Killer Cells, Natural , Leukocytes, Mononuclear , Liver NeoplasmsABSTRACT
Objective To explore the role and mechanisms of 5-HT1Areceptors in the medial septum-diagonal band of Broca complex (MS-DB) in hemiparkinsonian rats. Methods Combined behavioral and electrophysiological studies were performed to assess the role of MS-DB 5-HT1Areceptors in working memory and hippocampal theta rhythm in rats with 6-hydroxydopamine (6-OHDA)lesions in the medial forebrain bundle (MFB).Results ① MFB lesions in the rats decreased choice accuracy in the T-maze rewarded alternation test. Intra-MS-DB injection of 5-HT1Areceptor agonist 8-OH-DPAT further decreased choice accuracy,while intra-MS-DB injection of 5-HT1Areceptor antagonist WAY-100635 increased choice accuracy in the lesioned rats.② MFB lesions in the rats decreased peak theta rhythm frequency. Intra-MS-DB injection of 8-OH-DPAT suppressed hippocampal theta rhythm and decreased normalized theta power,while intra-MS-DB injection of WAY-100635 induced theta rhythm and increased normalized theta power.Conclusion Blockade of MS-DB 5-HT1Areceptors can recover cognitive dysfunction in Parkinson's disease, which may be attributed to the enhancement of hippocampal theta rhythm.
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Objective To investigate the efficacy and safety of clopidogrel hydrogen sulfate plus nimodipine in treating transient cerebral ischemic attack.Methods A total of 159 outpatients and inpatients with transient cerebral ischemic attack in the neurosurgery department and the neurology outpatient department of our hospital from January 1,2012 to September 1,2016 were divided into 3 groups.The group A(43 cases) orally took clopidogrel hydrogen sulfate plus aspirin;the group B (51 cases) treated with oral clopidogrel hydrogen sulfate plus nimodipine;the group C(65 cases) was treated with clopidogrel hydrogen sulfate plus nimotop.The curative effects were observed in 3 groups.The differences of PT.APTT,FIB and PLT in 3 groups were compared between before and after treatment.Results The total effective rate of the group A and C was significantly better than that of the group B(P0.05);the effect of the group B and group C on coagulation was superior to that of the group A(P<0.05).The three group had no severe bleeding events(cerebral bleeding,etc.),the difference was statistically significant.Conclusion Clopidogrel hydrogen sulfate combined with nimotop can significantly reduce the frequency of transient cerebral ischemic attack,has little influence on the coagulation indicators,and is safe and effective.
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Background and purpose:In recent years, the studies indicated that postoperatively induced myeloid-derived suppressor cells (MDSCs) were qualified with potent proangiogenic and tumor-promotive ability. Bone marrow mesenchymal stem cells (BMSCs) significantly inhibited the induction and proliferation of MDSCs. However, the relationship of MDSCs and tumor metastasis during perioperative period, and whether BMSCs could prevent tumor metastasis through inhibiting MDSCs are not clariifed. This study aimed to investigate the change of MDSCs during perioperative period and its correlation with tumor metastasis after surgery, and the inlfuence of BMSCs on the induction of MDSCs and the development of postoperative tumor metastasis.Methods:LLC cells were injected intravenously into C57BL/6 mice. Two hours later, these mice were divided into 4 groups: controls (C group); mice given anesthesia (A group); mice given anesthesia and laparotomy (AL group) and mice given anesthesia, laparotomy, and hepatic lobectomy (ALH group). The AL mice were divided into 2 groups after surgical operation: the AL mice without treatment (ALL group) and the AL mice treated with syngeneic BMSCs (ALB group). The percentage of Gr-1+CD11b+ cells in peripheral blood mononuclear cells (PBMCs) was detected by flow cytometry. The numbers of metastases on the lung surface were counted on the 14th day after LLC infusion. BMSCs were also co-culturedin vitro with myeloid cells in order to illustrate the effects of BMSCs on the generation of MDSCs.Results:The numbers of lung metastases in AL and ALH group signiifcantly increased as compared with C and A group (P<0.01). The number of lung metastases in ALH group signiifcantly increased as compared with AL group (P<0.05). The percentage of Gr-1+CD11b+ cells in PBMCs during postoperative period signiifcantly increased in AL and ALH group as compared with C and A group, and the percentage of Gr-1+CD11b+ cells in ALH group also signiifcantly increased as compared with AL group. The numbers of lung metastases in AL and ALB group were (38.00±9.57) and (6.54±1.49), the difference was statistically signiifcant (P<0.01) on day 14 after LLC infusion. Meanwhile, the percentage of Gr-1+CD11b+ cells in ALB group signiifcantly decreased as compared with AL1 group. This study also demonstrated that BMSCs inhibited the induction and proliferation of MDSCs from myeloid cells in vitro.Conclusion:Surgery stress induces MDSCs and promotes tumor metastasis. Syngeneic BMSCs could inhibit the generation of MDSCs and further suppress tumor metastasis after surgery.
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The paper was aimed to establish a quality evaluation model for Gualou Guizhi decoction based on the chemical compositions and biological effects. Ultra high performance liquid chromatograph-mass spectrometer was used to analyze and determine 24 kinds of chemical compositions in Gualou Guizhi decoction, and then, biological activity effect was quantitatively assessed in a zebrafish neuronal injury model which was induced by mycophenolate mofetil (MMF). As a result, the established method for quality evaluation of Gualou Guizhi decoction based on the chemical compositions and biological effects is feasible, stable and reliable, which can provide reference for quality control of compound Chinese medicines.
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Primary liver cancer is a common malignant tumor in the digestive system.Most patients are in the advanced stage when a con-firmed diagnosis has been made,and prognosis is often unfavorable.Traditional Chinese medicine (TCM)has unique advantages in the pre-vention and treatment of primary liver cancer.This article briefly introduces the etiology and pathogenesis of primary liver cancer,as well as syndrome differentiation and treatment of liver cancer with TCM,and summarizes the advances in multimodality therapies including TCM combined with transcatheter arterial chemoembolization,radical surgery for liver cancer,chemoradiotherapy,and molecular-targeted thera-py,in order to provide a reference for the clinical treatment and improvement in the prognosis of primary liver cancer.
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Objective To examine the expression of transforming growth factor β(TGF-β) activated kinase 1 (TAK1) in esophageal organization and the impact of TAK1 expression on clinicopathologic data. Methods Specimens from 80 patients with esophageal adenocarcinoma managed in our hospital were included in this study. Immunohistochemical staining was used to detect the expression of TAK1 in 80 cases of esophageal squamous cell carcinoma tissues and 80 cases of normal esophageal mucosal tissues. All the specimens were confirmed by pathology for esophageal squamous cell carcinoma or esophageal normal tissues. Results The positive expression rate of TAK1 in esophageal cancer organizations and normal esophageal mucosal tissues were 80% and 11.25%, respectively. The positive expression rate of TAK1 in esophageal cancer were significantly higher than those in normal esophageal mucosal tissue (P 0.05), but with lymph node metastasis and clinical stage (P<0.05). Patients with positive TAK1 expression had significantly lower five-year survival rate than those with tumors having positive TAK1 expression Conclusion TAK1 played an important role in the pathology and development of squamous cell carcinoma , and could be an important therapeutic target in the treatment of esophageal cancer.
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OBJECTIVE: To establish the HPLC fingerprint of Semen Strychni by chemometrics and the method of linear calibration using two reference substances (LCTRS). METHODS: The fingerprint of Semen Strychni was established by HPLC, using similarity analysis, 2-dimensional clustering analysis and principal component analysis for quality evaluation. LCTRS was used for identification of chromatographic peak. RESULTS: Fourteen chromatographic peaks were selected as common peaks in the fingerprints of 10 batches of Semen Strychni, and the chromatographic peaks of loganic acid, chlorogenic acid, strychnine and brucine were selected as characteristic peaks. By means of chemometrics evaluation, 10 batches of Semen Strychni were classified by their origins. The precision of predicting retention time by LCTRS was superior to the relative retention time method. CONCLUSION: LCTRS is more accurate for predicting retention time and suitable for more kinds of chromatographic columns than relative retention time method. In addition, the established fingerprint is specific, and can be used for the quality evaluation of Semen Strychni.
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Objective To investigate the effect of PCIA with low-dose fentanyl on plasma endothelin (ET) after craniotomy. Methods 47 cases of selected craniotomy were divided randomly into two groups :experimental group (26cases) and control group (21cases) . Patients in experimental group were treated with PCIA (fentanyl 15ug/kg +ondansetron 8 mg+100 mL NS) but patients in control group were not given PCIA. Then HR,MBP, VAS, ET and side-effects were observed and compared between two groups before treatment and 0, 2, 4, 8, 12, 24, 48 h after treatment.Results In experimental group, HR was lower at 2, 4, 8, 12, 24 and 48 h after treatment than control group. MBP was lower at 0 and 2h after treatment in experimental group than control group. Plasma levels of ET were lower at 8, 24 and 48h after treatment in experimental group than control group. There were significant differences in VAS scores at 2,4,8,12,24 and 48 h after treatment between two groups. There was no significant differences in side reactions including consciousness, respiratory depression, vomiting and sedation between two groups. The incidence of nausea was higher in experimental group than control group. Conclusion PCIA with low-dose fentanyl after craniotomy has good analgesic effect and few side reactions, can reduce the formation of plasma endothelin, and then alleviate brain damage.
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Objectives:To investigate the effects of amylin on bone minernal density and structure parameters of bone tissue in glucocorticoid induced osteoporosis rats. Methods:Four groups of female Wistar rats (3 months old) were treated for 12 weeks as follows: ⅠNormal Control, ⅡDXM, Ⅲ DXM+AMY;Ⅳ DXM+Vitamin Da 3. By intramuscular injection of dexamethasone(XM)1mg/kg twice a week during the first 8 weeks, the animal model of Glucocorticoid induced osteopoprosis was established. After 12 weeks, BMD of the lumbar vertebrae and the femural bone were measured by DEXA. The bone morphology of the lumbar vertebrae was also determined. Results:①After the treatment with AMY, bone mineral density (BMD) was significantly increased at the lumbar spine and the femural bone. ( vs. DXM group, P
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Objective To obtain Chinese hamster ovary (CHO) cell lines that stably express a targeting complement inhibitor CR2-CD59.Methods The recombinant plasmid PEE14.1-CR2-CD59 was constru-cted by cloning the DNA fragment CR2-CD59 into plasmid PEE14.1,and the obtained plasmid was transfected into CHO cells by FuGENE 6.The clones with stable high expression of target fragment were selected by methionine sulfoximine (MSX),the expression of CR2-CD59 was analyzed by ELISA,SDS-PAGE and Western blotting analysis.Results Several stable expression clones were obtained,and CR2-CD59 was highly expressed in the secret form in CHO cells.SDS-PAGE analysis showed that the molecular weight of the recombined protein CR2-CD59 was consistent with the predicted one.ELISA and Western blotting results revealed that the CR2-CD59 could react with both anti-human CR2 and anti-human CD59 polyclonal antibodies.Compared with serum-containing medium,the protein was highly expressed in serum-free medium (P
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Objective To observe and compare the effects of four kinds of anesthetic methods on the revival time,the rate of re-dormant after revival,the total Ketamine's doses and the respiratory status after extubation,and to improve the safety of anesthesia.Method 80 cases of children with palatorrphy were randomly divided into 4 groups: group A(n=20) given Midazolam and Ketamine,group B(n=20) given Fentanyl and Ketamine,group C(n=20) given Isoflurane and Ketamine,and group D(n=20) given Sevoflurane and Ketamine.Results(1) The revival time in group D and group C shortened(D and C