The Capping Protein β-Subunit Mediates Fusicoccin-induced Stomatal Movement in Arabidopsis / 中国生物化学与分子生物学报

Actin dynamics in guard cells play a critical role in stomatal movement. Remodeling of actin arrays is triggered by different biotic and abiotic stimuli, which requires precise control. However, the molecular mechanism underlying this process is not well understood. Here we investigated whether and how the capping protein (CP) regulates actin filaments during fusicoccin (FC) -induced stomatal opening. We found that both stomatal opening and F-actin rearrangement are sensitive in the Capping Protein β-subunit (CPB) cpb-3 mutants, which resulted in its hypersensitivity to drought stress. The leaves detached from cpb-3 had a higher water loss rate (63. 45%) than from the wild type (48. 99%), and the stomatal aperture of cpb-3 was about 20% greater than in the wild type. After 1 h of FC treatment, the proportion of cpb-3 guard cells with radial actin arrays increased to 65. 5% dramatically, while only approximately 47. 2% guard cells in WT exhibited transversely oriented actin filaments. Moreover, the record of transmembrane Ca

Nanocarrier-mediated PiggyBac transposon system for preparation of CAR-NK cells / 中国肿瘤生物治疗杂志

@#Objective: To explore the gene transduction method of chimeric antigen receptor (CAR) mediated by novel cationic polymer nanocarrier mPEG-P (Asp-AED-g-HFB) (PAEF) and PigyBac transposon system to modify natural killer (NK) cells, providing a new strategy for immunotherapy of cancer cells. Methods: PAEF/DNA (transposase+transposon) complex were prepared. The particle size distribution and surface potential of PAEF/DNA complexes were measured with Nano-ZSE Dynamic Light Scattering System (Malvern Instruments). The DNA encapsulation rate, release and stability of PAEF were evaluated by DNA gel electrophoresis, and then by combiningwithparticlesizeandsurfacepotentialtodeterminethepreferentialN/PratiotoenterNKcells.Thecell cytotoxicity of PAEF/DNA complexes under different N/P ratios was analyzed by CCK-8 cytotoxicity test. Transduction efficiency of NK cells was evaluated by Fluorescence microscopy and Flow cytometry, and the feasibility of PAEF gene transfection vectors was assessed. Results: PAEF could encapsulate DNA to form nano-complexes with the diameter of 100-150 nm, which was suitable to mediate DNA entering into cells. PAEF could completely encapsulate DNA with N/P ratio of 20. In the presence of reducing agent dithiothreitol (DTT), PAEF had a good ability to release DNA. NK-92 cells transfected with PAEF/DNA complex, which was formed at the N/P ratio of 80, attained a significantly higher cell viability than cells of lipofectamine transfection group [(72.50±3.9)% vs (64.03±1.8)%, P<0.05]; Fluorescence microscopic observation showed more fluorescence and higher fluorescence intensity in cells of PAEF/DNA group; Flow cytometry showed the highest transfection efficiency of 83.4%. Conclusions: Nanocarrier PAEF can encapsulate DNA well by electrostatic adsorption, and has good biocompatibility and high efficiency for gene transduction. It provides a good experimental basis for adoptive immunotherapy.

Protective role of brain CYP2J in diverse Parkinson disease animal models / 中国药理学与毒理学杂志

OBJECTIVE CYP2 family including CYP2C and CYP2J is the predominant arachidonic acid (AA) epoxygenase, and the epoxidation of AA produces four regioisomeric cis-epoxyeicosatrienoic acids (5,6-, 8,9-, 11,12-, and 14,15-EET). Human CYP2J2 is one of the main CYP isoforms expressed in brain, but CYP2C8 was present at a low level. The aim of this study is to investigate the roles of brain CYP2J in Parkinson disease. METHODS Rats received the right-unilaterally injection with concentrated LV-CYP2J3 or LV-EGFP in the substantia nigra (SN) at 3 d before LPS or 6-OHDA treatment. The animals were tested for rotational behavior with the dopaminergic agonist apomorphine dissolved in sterile saline at 14 and 21 d after LPS injection. The influence of CYP2J-dependent derivative, 14,15-EET, on the genes related with oxidative stress was assayed in SH-SY5Y cells. RESULTS CYP2J overexpression or 14,15-EET treatment significantly increased the levels of SOD1, CAT, GPX1, NRF2 and KEAP1 in neurons. TLR4- MyD88 signaling pathway was involved the down- regulation of CYP2J by LPS. The binding of p-CREB with the promoter of CYP2J was inhibited by the LPS treatment. The loss of dopami?nergic neurons in the right SN induced by LPS or 6- OHDA was significantly decreased by CYP2J3 transfection at 21 d after LPS injection. Compared with LPS or 6-OHDA group, the number of the rotation of rats was decreased by 42.6% and 60.7% by CYP2J3 transfection at 14 d after LPS or 6-OHDA injection;meanwhile, the rotation number was decreased by 12.7% and 21.3% at 21 d. The accumulation of alpha synuclein induced by LPS was significantly decreased by CYP2J3 transfection. The mRNA levels of SOD1, CAT, GPX1, NRF2 and KEAP1 in SN were decreased by LPS, which was attenuated by the injection of LV-CYP2J3. CONCLUSION Brain CYP2J can play a protective role in the damage of the inflammation and oxidative stress to the dopaminergic neurons. Brain CYP2J- dependent derivatives from AA may have therapeutic effects in Parkinson disease via the up- regulation of the antioxidant system in neurons.

An analysis for the phenotype and genotype of autosomal dominant polycystic kidney disease from two Chinese families / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To scan for mutations of polycystic kidney disease 1 gene (PKD1) in Chinese population in order to find some features about Chinese patients and a better approach to detect mutations.</p><p><b>METHODS</b>Twenty-five PKD-affected individuals from twenty-one unrelated genealogies and sixteen controls participated in the study. Thirty-five blood samples and six tissues were obtained after receiving informed consent and were in accordance with institutional ethical guidelines. Genomic DNA was isolated from peripheral blood using standard procedures. PCR amplification of genomic DNA was performed to generate the aimed fragments. Amplified fragments were analyzed by denaturing gradient gel electrophoresis (DGGE). A GC clamp was attached to the 5' primer. After that, the abnormal fragments were sequenced on freshly amplified specific PCR products with the dideoxynucleotide chain termination method. Sequencing was performed for all samples to evaluate DGGE.</p><p><b>RESULTS</b>Aimed fragments of exons 44 and 45 were amplified. DGGE detected eleven abnormal PCR fragments. Two novel mutations were identified by sequencing, included one nonsense mutation (C12217T) and one frameshift (12431delCT). In addition, one polymorphism (A50747C) was identified. The mutation detection rate is 8% in our study.</p><p><b>CONCLUSION</b>Two novel pathogenic mutations were detected, including one nonsense mutation (C12217T) and one frameshift (12431delCT).</p>

Cloning and expression of human interleukin-26 in Escherichia coli / 生物工程学报

To clone human interleukin-26 (hIL-26) and express it in E. coli efficiently. Two pairs of primers were synthesized according to the hIL-26 gene reported on GenBank. The hIL-26 gene was cloned by nest PCR following the first round RT-PCR from human peripherial blood monocytes total RNA, and then the PCR product was cloned into pMD18-T vector. Colony PCR, restriction analysis and sequence analysis showed that the gene cloned was the same as the reported hIL-26. The recombinant was cut with BamHI and EcoR I to obtain the hIL-26 fragment, and then the fragment was inserted into pBV220 which was cut with the same enzymes. The recombinant expression vector was induced to express hIL-26 at 42 degrees C, SDS-PAGE analysis showed that the recombinant protein accounted for up to 20% of the whole protein of E. coli, and the protein was also confirmed by Western blotting. Purity of the protein was found to be above 90% after purified with molecular sieve. After renaturalized with glutathione buffer, the promoting effect of it on the production of IFN-y in PBMC was detected by RT-PCR. A recombinant bacterial strain for expressing hIL-26 with biological activity was constructed successfully.

Clinical observation on effect of shenqi fanghou recipe in preventing and treating radiation injury in patients with head and neck tumor / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the therapeutic effect of shenqi fanghou recipe (SFR) in preventing and treating radiation injury in patients with head and neck tumor.</p><p><b>METHODS</b>One hundred and forty patients with head and neck tumor, including nasopharyngeal carcinoma, carcinoma of tonsil or tongue, were randomly divided into 2 groups, 70 patients in the observed group were given modified SFR as adjuvant to radiotherapy, while 70 patients in the control group were treated with radiotherapy alone. The radiation reactions during radiotherapy and the condition of late stage radiation injury radiotherapy in patients in the 2 groups were observed.</p><p><b>RESULTS</b>The degree of oropharyngeal mucosa reaction, dryness in mouth and radiation dermatitis in cervical region in the observed group was milder than those in the control group, and the radiation injury induced late stage sequelae, such as the degree of mouth-opening was better and the cervical muscular sclerosis was better in the observed group than in the control group, showing significant difference (P < 0.01).</p><p><b>CONCLUSION</b>SFR has definite effect in preventing and treating radiation reaction and late stage radiation injury in patients with head and neck tumor.</p>

Study on the (TAAAA)n repeat polymorphism in sex hormone-binding globulin gene and the SHBG serum levels in putative association with the glucose metabolic status of Chinese patients suffering from polycystic ovarian syndrome in Shandong province / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the relationship of (TAAAA)n repeat polymorphism in the promoter of the sex hormone-binding globulin (SHBG) gene and SHBG serum levels to the glucose metabolic status of Chinese polycystic ovary syndrome (PCOS) patients in Shandong province.</p><p><b>METHODS</b>GeneScan method was used to detect and identify (TAAAA)n repeat number (alleles) and genotypes for 156 controls and 157 patients who were divided into normal glucose tolerance without hyperinsulinemia (NIR group) and with hyperinsulinemia (HI group) and abnormal glucose metabolic (AGM) group according to the results of oral glucose test and insulin resistant test; IRMA was used to measure serum SHBG for part of them.</p><p><b>RESULTS</b>Five alleles containing (TAAAA) 6-10 repeats and 14 genotypes including 6/6, 6/7, 6/8, 6/9, 6/10, 7/7, 7/8, 7/9, 7/10, 8/8, 8/9, 8/10, 9/9, 9/10 repeats genotypes were present in the subjects. Genotype distribution of 6/10 repeats genotype is lower in PCOS than that in control, and 8/9 repeats genotype vice versa (P < 0.01); among PCOS subgroups, the eight repeat genotypes in NIR group is more frequent than that in HI group (P < 0.01), and 7/9 genotype distribution in AGM group is higher than that in NIR group and HI group(P < 0.05-0.01). The serum SHBG levels in homozygous genotype groups exhibit a sequence of 8/8 > 9/9 > 6/6, 7/7 repeats and the fall of serum SHBG trend is in reversed relation with the increase in body mass index (BMI), Homa-IR, and blood pressure. Serum SHBG levels in AGM exhibit a sequence of HI group < NIR group < control but show no statistical difference between both groups.</p><p><b>CONCLUSION</b>This study reveals that the repeat number, alleles, genotypes and their distributions in Chinese women are very different from these in foreigners. Some special genotypes and low serum SHBG levels may be associated with PCOS and its glucose metabolic status; some special genotypes may influence Chinese serum SHBG and need more studies, but both SHBG gene polymorphism genotype and serum SHBG are not good indicators to find out the PCOS individual at high risk.</p>

Influence of chemotherapy on Th1/Th2 cytokine switching in stomach cancer patients / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To observe the influence of chemotherapy on the switching of Th1/Th2 cytokines in stomach cancer patients.</p><p><b>METHODS</b>Th1/Th2 cytokine genes expressed by peripheral blood mononuclear cells of stomach cancer patients before and after chemotherapy were detected by RT-PCR.</p><p><b>RESULTS</b>The expression of Th2 cytokines was dominant in patients before chemotherapy, and the dominancy became less marked after chemotherapy.</p><p><b>CONCLUSION</b>The immune deviation with Th2 predominance in stomach cancer patients has a tendency to become reversed after chemotherapy.</p>