雷公藤红素白蛋白纳米粒的制备及抗类风湿性关节炎药效评价

目的制备雷公藤红素白蛋白纳米粒(CLT-AN),并考察其体内抗类风湿性关节炎(RA)活性。方法采用超声法制备CLT-AN,以粒径、分散指数(PDI)和稳定性为评价指标,以CLT投药量、大豆油用量和超声功率为考察因素,通过单因素实验优化处方工艺。利用透射电镜、激光粒度测定仪等考察CLT-AN的理化性质,并研究其体外稳定性和体外释放特性。构建佐剂诱导关节炎大鼠模型,考察CLT-AN对模型大鼠关节肿胀、血清中炎症因子[肿瘤坏死因子α(TNF-α)和白细胞介素1β(IL-1β)]水平以及关节组织病理状态的影响。结果优化的处方为CLT 6.5 g、大豆油45 mg、超声功率490 W、超声时间8 min。按最佳处方制备的CLT-AN外观形态圆整,平均粒径为(96.8±1.1)nm,PDI为0.174±0.020,Zeta电位为(-18.6±1.7)mV,包封率为(94.61±0.46)%,载药量为(2.42±0.21)%。在室温条件下,CLT-AN储存5 d内的粒径、PDI、Zeta电位和包封率均无显著变化。CLT-AN体外释放缓慢,72 h的累积释放量可达到73.56%;与游离CLT相比,CLT-AN可明显抑制模型大鼠的关节肿胀,降低其血清中TNF-α、IL-1β水平,并且显著改善炎症关节组织的病理状态。结论超声法制备的CLT-AN粒径适中、稳定性良好,具有显著的缓释特征和优良的抗RA活性。

我们的素白华年

我们的故事，其实没有开始，也没有后来。10年之后，若是我重新回到那个当初奋斗过的播音筒前；10年之后，等到我们都已长大，大得年龄都不适合上演王子和公主的童话了，等到我们都只剩下回忆的时候，我不知道，你是否还记得有这样一个搭档，曾毫无顾忌地叫你“美女”。

Effect of Foliar Fertilizer Spraying at Seedling Stage on the Growth Physiological Characteristics and Yield of Brassica napus L.

In order to select a suitable foliar fertilizer for Brassica napus L.at the seedling stage,using‘Fengyou 958’as the material,different foliar fertilizers including BR,Se,Si,BR+Si,BR+Se,Se+Si and BR+Se+Si were sprayed at the seedling stage to study their effect on the physiological characteristics,growth and yield of Brassica napus L..The results showed that the growth,chlorophyll content,soluble sugar content,soluble protein content,and yield of different treatments at the budding stage improved compared with the control.The effect of the Si+Se treatment was the best,followed by the Se and Si treatments.The chlorophyll content of the flower decreased continuously during the whole flowering period,and the chlorophyll content of the mature silique peel was higher than that of the seed.The content of soluble sugar in flowers was the highest in the early flowering stage,and the content of soluble sugar in leaves at the flowering stage was higher than that at the budding stage.The soluble sugar content in the harvested silique peel decreased gradually with the increase of time,and that in the silique peel and seed was similar at 35 d after pollination.The soluble protein content in the silique peel and seed decreased gradually at the mature stage,and the soluble protein content in the silique peel was higher than that in the seed at the same stage.Si+Se foliar fertilizer spraying at the seedling stage can promote the growth and yield of Brassica napus L.and can be applied in Brassica napus L.production.

Megestrol acetate plus metformin for fertility-sparing treatment of atypical endometrial hyperplasia and early-stage endometrial adenocarcinoma: a prospective study

Objective To evaluate the efficacy of medroxyprogesterone acetate(MA)plus metformin as the primary fertility-sparing treatment for atypical endometrial hyperplasia(AEH)and early-stage grade 1 endometrial adenocarcinoma(G1 EAC)and the recurrence rate after treatment.Methods Sixty patients(aged 20-42 years)with AEH and/or grade 1 EAC limited to the endometrium were enrolled prospectively and randomized into two groups(n=30)to receive oral MA treatment at the daily dose of 160 mg(control)or MA plus oral metformin(850 mg,twice a day)for at least 6 months.The treatment could extend to 12 months until a complete response(CR)was achieved,and follow-up hysteroscopy and curettage were performed every 3 months.For all the patients who achieved CR,endometrial expressions of IGFBP-rP1,p-Akt and p-AMPK were detected immunohistochemically.Results A total of 58 patients completed the treatment.After 9 months of treatment,23(76.7%)patients in the combined treatment group and 20(71.4%)in the control group achieved CR;two patients in the control group achieved CR after converting to the combined treatment.The recurrence rate did not differ significantly between the control group and combined treatment group(30.0%vs 22.7%,P>0.05).Ten(35.7%)patients in the control group experienced significant weight gain of 5.7±6.1 kg,while none of the patients receiving the combined treatment exhibited significant body weight changes.Compared with the control group,the patients receiving the combined treatment showed enhanced endometrial expressions of IGFBP-rP1 and p-AMPK with lowered p-Akt expression.Conclusion Metformin combined with MA may provide an effective option for fertility-sparing treatment of AEH and grade 1 stage IA EAC,and the clinical benefits of metformin for controlling MA-induced weight gain and promoting endometrial expressions of IGFBP-rP1 and p-AMPK while inhibiting p-Akt expression warrants further study.

Chloroplast Composition and Structural Differences in a Chlorophyll-reduced Mutant of Oilseed Rape Seedlings

对黄化突变体Cr352 9和野生型油菜 (BrassicanapusL .) 352 9叶绿体的超微结构和组成进行了比较。与野生型相比 ,突变体Cr352 9叶片具有较少的类囊体、较少的垛叠膜区和较少的叶绿素含量。突变体的Chla/Chlb比值较高 ,是野生型的 2倍。电泳结果表明 ,突变体类囊体膜中LHCⅡ和其三聚体LHCⅡ 的含量减少。SDS_PAGE分析显示 ,LHCⅡ的脱辅基蛋白在突变体类囊体膜中明显减少。免疫印迹进一步表明 ,所有LHCⅡ组分的含量仅为野生油菜的类囊体膜的 1 / 3。突变体Cr352 9的天线系统比野生型 352 9的小。

Expression analysis of ThGLP, a new germin-like protein gene, in Tamarix hispida

Germin and Germin-like protein （GLP） have various proposed roles in plant developmental stages and stress- related processes. A novel GLP cDNA clone was isolated from a cDNA library of Tamarix hispida. ThGLP, coded 225aa, possesses conserved motif of plant germin and Germin-like protein. ThGLP belongs to true germin subfamily through phylogenetic analyses. Gene expression profiles in roots and leaves were evaluated using real-time quantitative RT-PCR. The results show that the gene was highly induced by drought, salt, low temperature, CdCl2 and abscisic acid treatments. Our results demonstrate that the ThGLP gene is expressed in leaves and roots, is involved in different abiotic stress re-sponses and controlled by an ABA-dependent signaling pathway.

Qualitative and Quantitative Analysis of Rhizoma of Polygonum cuspidatum

Aim To establish reliable methods for evaluating the quality of rhizoma of Polygonum cuspidatum( Huzhang in Chinese). Methods TLC and HPLC were employed for the chemical identification and content determination,respectively. Results A qualitative TLC method and a quantitative HPLC method with piceid as the reference substance were established, respectively. With piceid as the reference substance and ethyl acetate-methanol-formic acid-water ( 19:3:0.5:1) as the mobile phase, a TLC method for the identification of Huzhang from the commonly used crude drugs of the same family was also set up. Conclusion The established TLC method can reasonably appraise the quality of the drug and easily distinguish Huzhang from the other commonly used crude drugs of the same family. The HPLC method for determining piceid is simple, reproducible, accurate, and feasible.

Characteristics and Comparative Study of Chlorophyll-protein Complexes from Siphonous Green Algae

By mild PAGE method, 11, 11, 7 and 9 chlorophyll_protein complexes were isolated from two species of siphonous green algae (Codium fragile (Sur.) Hariot and Bryopsis corticulans Setch.), green alga (Ulothrix flacca (Dillw.) Thur.), and spinach (Spinacia oleracea Mill.), respectively. Apparent molecular weights, Chl a/b ratios, distribution of chlorophyll, absorption spectra, low temperature fluorescence spectra of these complexes were determined, and compared with one another. PSⅠ complexes of two siphonous green algae are larger in apparent molecular weight because of the attachment of relative highly aggregated LHCⅠ. Four isolated light_harvesting complexes of PSⅡ are all siphonaxanthin_Chl a/b_protein complexes, and they are not monomers and oligomers like those in higher plants. Especially, the absence of 730 nm fluorescence in PSⅠ complexes indicates a distinct structure and energy transfer pattern.

Effects of High Light Stress on Chlorophyll-protein Complexes of Two Subspecies of Rice

Influence of high light stress on the photosynthesis of flag leaves of indica subspecies (cv. “Shanyou 63', sensitive to photoinhibition) and japonica subspecies (cv. “Wuyujing', resistant to photoinhibition) of rice ( Oryza sativa L.) was comparatively investigated. In both cultivars of rice, the excitation energy distribution between two photosystems was altered and the excitation energy transfer from light harvesting chlorophyll protein complexes to PSⅡ was inhibited by high light stress. These decreases were more pronounced in indica rice cultivar as compared to japonica one. The analysis of mild SDS_PAGE showed that in indica rice, high light stress almost disaggregated the trimer of light harvesting chlorophyll protein complexes of PSⅡ (LHC Ⅱ 1). The stress reduced the contents of internal antennae chlorophyll protein complexes of PSⅡ (CPa), light harvesting chlorophyll protein of PSⅠ (CPⅠa) and Chl a protein complex of PSⅠ reaction center (CPⅠ) as well as dimer of LHCⅡ (LHCⅡ 2) in indica rice. In japonica subspecies, however, high light stress depressed the contents of LHCⅡ 1, CPa and CPⅠa, but slightly impacted on CPⅠ content. Moreover, the increase in the contents of monomer of LHCⅡ by high light stress was found in both subspecies. In consistent with above results, analysis of polypeptide indicated that the amounts of 27 kD and 25 kD polypeptide of LHCⅡ in particular, as well as that of 21 kD polypeptide of CPⅠa were reduced by high light stress in both subspecies. It was found that, comparing with japonica rice, the stress pronouncedly diminished 43 kD and 47 kD proteins of CPa and 23 kD extrisic protein in indica rice.

Plant Anthocyanin Synthesis and Gene Regulation

This paper aims to explain the biochemistry of anthocyanin synthesis based on an overview of plant anthocyanin synthesis genes and environmental factors in the regulation of anthocyanin metabolism. The results show that： ① The metabolism of anthocyanins in plants is affected by the temperature, light, ultraviolet, fertilization status, hormone levels and other factors, which affect the military anthocyanin biosynthetic genes, and then induce or inhibit the synthesis of anthocyanins. ② In the regulation of genes, some of the structural genes of anthocyanin synthesis showed promoting effect, while others showed inhibitory effect. At different environ- mental conditions, the regulation of gene activation and inhibition of the amount of different regulatory genes that anthocyanin accumulation is different, and cause different colors of plant-organs production. ③ In different environmental factors or hor-mones induced to produce the same or different regulation of gene expression changes in regulatory genes, resulting in several different anthocyanins or anthocyanin ratio changes, so that the color of plant organs in different colors.

Changes of Chorophyll protein Complexes and Photosynthetic Activities of Chloroplasts from Lotus (Nelumbo nucifera) Seeds Germinating in Light

The changes of chlorophyll_protein complexes and photosynthetic activities of chloroplast isolated from lotus ( Nelumbo nucifera Gaertn.) seeds germinating under illumination were studied. SDS PAGE analysis of chlorophyll_protein complexes showed that there was only the light harvesting chlorophyll a/b protein complex from PSⅡ (LHCⅡ) precursor in chloroplast from lotus seeds germinated for 2 to 6 days, while LHC Ⅱ 1, and the chlorophyll_protein complex of PSⅠ (CPⅠ) appeared on the 8th day of germination and PSⅡ reaction center complex appeared later. Studies on the polypeptides composition of the chloroplast revealed the following results: 1) Small amount of the 27 kD polypeptide was synthesized in invisible light; 2) The 30 kD polypeptide existed previously in the plumules of the dry seeds; 3) The amount of the 30 kD polypeptide was more than any other polypeptides before germination and decreased gradually throughout germination, while the 27 kD polypeptide changed in the opposite way; 4) In the process of germination, measurement of the electron transport rate and the fluorescence induction kinetics at room temperature showed that PSⅡ activities and efficiency of primary light energy transformation were only experimentally measurable after 7 days of germination and gradually increased afterwards. At the same time, the chl a/b ratio rose from the lower value to normal; 5) The changes of chloroplast membrane components and its functions are concomitant in concert with that of the ultrastructure of chloroplast membranes during germination, as shown in our earlier work . The results have proved again that a different developmental pathway of chloroplast is likely to exist in the lotus plumules, which might provide an important clue for N. nucifera in having an unique position in the phylogeny of the angiosperm.

Gel Filtration Chromatography Combined with Bradford Method for Determination of Total Residual Protein in Ferment Antibiotics

Aim A novel method has been developed for evaluation of the levels of total residual protein in antibiotics produced by fermentation using gel filtration chromatography (GFC) combined with Bradford assay based on determination of residual protein in lincomycin hydrochloride. Methods The chromatographic conditions were SuperdexTM peptide column, 0.01 mol*L-1 phosphate buffer solution as mobile phase, and flow rate of 1 mL·min-1. Five hundred microliters of lincomycin hydrochloride solution (3 g of lincomycin hydrochloride dissolved in 10 mL of mobile phase) was injected into the chromatograph and the eluted solution was collected between 6 min and 14.5 min (protein eluted from column within this period), and the residual content of total protein in the eluted solution was assayed using Bradford assay method. Results The average recovery was more than 90% for bovine serum albumin, the calibration equation for the range of 0-12 μg·mL-1 of protein was y=-0.002 4x2+0.064 2x+0.002 9, r2=0.999 9, RSD=0.1%-0.9%, and the LOD and LOQ were 3 and 10 ng·mL-1 of protein, respectively. Conclusion The novel method for determining the residual protein in ferment antibio-tics is simple, rapid, and precise.

The in vitro Anti Hptocarcinoma Effects of Human Interleukin-12 from Transgenic Potato

[Objective]The aim was to examine the anti heptocarcinoma effects of human interleukin-12（hIL-12） from transgenic potatoes.[Method]Human heptocarcinoma cell line HepG22.2.15 was cocultured with human peripherial blood monocyte（PBMC）.Human IL-12 extracted from its transgenic patatoes was introduced to this coculture system.MTT method and laser confocal microscope were then employed to evaluate its survival rates and morphological changes of HepG22.2.15 cell line.[Result]The HepG22.2.15 survival rate of the plant produced hIL-12 treated group was significantly lower than that of the wild type control,while similar to that of commercial purified recombinant hIL-12 group.As indicated by AnnexinV/PI double staining laser confocal microscope,cocultured heptocarcinoma cells showed the typical early and final phase apoptotic morphological characteristics 48 hours after 2 × 10-4 mg/L potato secreted hIL-12 treatment.[Conclusion] These results demonstrated that Heptocarcinoma HepG22.2.15 cell growth could be actively inhibited by the transgenic potato expressed hIL-12 in vitro.

Alzheimer’s disease:epidemiology,genetics,and beyond

Alzheimer＇s disease （AD） is an increasing epidemic threatening public health. Both men and women are susceptible to the disease although women are at a slightly higher risk. The prevalence of AD rises exponentially in elderly people from 1% at age of 65 to approximately 40%-50% by the age of 95. While the cause of the disease has not been fully understood, genetics plays a role in the onset of the disease. Mutations in three genes （APP, PSENI, and PSEN2） have been found to cause AD and APOE4 allele increases the risk of the disease. As human genomic research progresses, more genes have been identified and linked with AD. Genetic screening tests for persons at high risk of AD are currently available and may help them as well as their families better prepare for a later life with AD.

Study on Fermentation Conditions of Pycnoporus sanguineus

[Objective] The purpose was to study the optimum conditions of Pycnoporus sanguineus for producing ligninase. [Method] A strain of lignindegrading white-rot fungus was selected from 5 strains of collected fungi and its ligninase production and the optimum conditions for producing ligninolytic enzyme were measured. [Result] It could produce two kinds of ligninase with good thermal stability. Different temperatures, carbon sources, nitrogen sources, acidities, as well as the additions of surfactant had distinct influence on the development of lignin-degrading enzymes of the fungus. The optimum condition was drawn out：38℃, pH = 4.5, 10.0 g/L glucose, 1.0 g/L tartaric acid ammonium. [Conclusion] The aim of research was to provide a basis for lignin degradation in practical production.

Study on Hardwood Cutting Propagation Technology of Fraxinus

[Objective] Hard-branch cutting propagation technology of Fraxinus was studied under different environmental conditions. [Method] An orthogonal test was designed based on 1-year-old clones of F. pennsylvanica Marsh Lula 5. Under dif- ferent growth hormones, hormone concentrations and processing time, F. velutina Lula 2 and F. pennsy/vanica Marsh Yuanla 2 were treated by vacuum pump for 10 min, [Result] The decreasing order of cutting survival rates of Lula 5 under different etnvironments was small plastic flowerpot （94.4%）, nutrition cup （92.8%）, sand tray （56.1%）, and cuffings treated with 100 mg/L ABT, for 2 h, 200 mg/L ABT, for 4 h and 100 mg/L IBA for 6 h under large nutrition cup environment grew the best, showing a survival rate of 100%. Lula 2 and Yuanla 2 were subjected to vacuum pump treatment, the survival rate of cuttings under the sand tray environment was averagely 29%; and under the large plastic flowerpot environment, the survival rate of cuttings was averagaly 64.4%, and cuttings of Lula 2 treated with vacuum pump for 10 min in 200 mg/L ABTI showed a survival rate up to 95%, which was the highest among all treatments. It could be seen that whether cuttings were treated with vacuum pump, the survival rate of cuttings in sand tray was lower than those under other environments, while there was no big difference in survival rate of cut- tings between the nutrition cup and plastic flowerpot environments. [Conclusion] This study is of great significance to improvement of cutting propagation coefficient.

Pharmacokinetics of Ivermectin in Salvelinus leucomaenis Following Two Ways of Administration

[Objective] This study was aimed to study the pharmacokinetics of iver- mectin in Salvelinus leucomaenis following a single oral administration or intraperi- toneal injection. [Method] After a single oral administration or intraperitoneal injection of ivermectin with a dosage of 0.3 mg/kg body weight, samples were taken at dif- ferent time points and determined by high-performance liquid chromatography-ultravi- olet detection （HPLC-UV）. The pharmacokinetic parameters were calculated by 3p97 software. [Result] The concentration-time relations of ivermectin in plasma, muscle, liver and kidney were well described by one-department open model with first-order absorption. After oral administration, the pharmacokinetic parameters in plasma were as follows： Tmax=4.503 h, Cmax=0.252mg/L, t1/2ka=0.476 h, t1/2ka=331.160 h, AUC=121.524 （mg/L） .h. After intraperitoneal injection, the pharmacokinetic parameters in plasma were as follows： Tmax=2.751 h, Cmax=0.230 mg/L, t1/2ka=0.306 h, t1/2ka=153.868 h, AUC= 51.689 （mg/L） .h. [Conclusion] There were differences in the pharmacokinetics of ivermectin in Salvelinus leucomaenis between two different administration routes, and intraperitoneal injection led to more rapid absorption than oral administration.

RICE TRANSFORMATION WITH A PHYTOALEXIN GENE AND BIOASSAY OF THE TRANSGENIC PLANTS

Immature embryos, mature embryos and embryogenic calli of 6 rice (Oryza sativa L.) materials were transformed with particle bombardment. The plasmids pSSVstl and pVE5+ were used, both containing the phytoalexin gene from grapevine coding for stilbene synthase, but driven by 35S and its own promoter respectively. Through resistance selection for G418 (100 to 150 mg/L) or hygromycin (50 mg/L), 54 independent transgenic plants were isolated and further assessed by PCR, Southern blot and Dot blot analyses. The transgenic plants and their progenies were tested for resistance to blast ( Pyricularia oryzae ) and bacterial blight of rice ( Xanthomonas oryzae ). Preliminary results indicated that the stilbene synthase gene could enhance the resistance of transgenic plants and their progenies to both pathogens.

Effects of Matrine on Adenine-Induced Chronic Tubulointerstitial Fibrosis in Rats

Aim ,To study the mechanism relative to therapeutic effects of matrine on adenine-induced renal interstitial fibrosis in rats. Methods Sixty male Wistar rats were selected and randomly divided into 3 groups： normal control group （NCG） consisted of 8 rats, adenine treated group （ATG） 28 rats, and matrine treated group （MTG） 24 rats. Each rat in ATG and MTG was gavaged with adenine （250 mg·kg^-1·d^-1 ） for 21 d. After gavage with adenine for one week, each rat in MTG was administered intraperitoneally matrine（20 mg·kg^-1·d^-1 ） in vehicle （ 1 mL of 0.9% sodium chloride） daily. On days 14, 21, and 28, the serum levels of urea nitrogen, creatinine, and IL-6 were determined and the rat kidneys of ATG, MTG and NCGwere examined pathologically. Radioimmunoassay for serum IL- 6 immunohistochemical staining for TGF-β1 expression in the kidney and semiquantitative analysis were performed. HE staining for semiquantitative analysis of tubulointerstitial injury. Results The serum levels of urea nitrogen and creatinine in MTG were lower as compare to ATG （ P 〈 0.05 ） whereas serum IL- 6 and renal TGF-β1 expression levels were significantly lower than those in ATG （P 〈0.05 ）, but all these indexes were higher than those in NCG （P 〈 0.01 ）. In MTG, the index of tubulointerstitial lesion was lower than that in ATG （P 〈 0.05 ）. Conclusion Matrine inhibits the renal tubulointerstial fibrosis in the adenine-induced rat model by suppressing serum level of IL-6 and expression of TGF-β1 in the tubulointerstitium.

谈留青竹刻的徐氏流派

留青竹刻是中华民族特有的传统民间艺术 ,江苏常州的留青竹刻在国内独树一帜。文章就常州留青竹刻大家徐素白及其后人徐秉方、徐秉言的风格与作品作了高度评价 ,也兼论了另一个竹刻家白士风及其传人的留青竹刻作品。