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Vascular dementia(VD) is a condition of cognitive impairment due to acute and chronic cerebral hypoperfusion. The available therapies for VD mainly focus on mitigating cerebral ischemia, improving cognitive function, and controlling mental behavior. Achievements have been made in the basic and clinical research on the treatment of VD with traditional Chinese medicine(TCM) active components, including Ginkgo leaf extract, puerarin, epimedium, tanshinone, and ginsenoside. Most of these components have anti-inflammatory, anti-apoptotic, anti-oxidant, and neuroprotective effects, and puerarin demonstrates excellent performance in mitigating cholinergic nervous system disorders and improving synaptic plasticity. Puerarin, ginkgetin, and epimedium are all flavonoids, while tanshinone is a diterpenoid. Puerariae Lobatae Radix, pungent in nature, can induce clear Yang to reach the cerebral orifices and has the wind medicine functions of ascending, dispersing, moving, and scurrying. Puerariae Lobatae Radix entering collaterals will dredge blood vessels to promote blood flow, and that entering the sweat pore will open the mind, which is in line with the TCM pathogenesis characteristics of VD. This study reviews the progress in the mechanism of puerarin, the main active component of Puerariae Lobatae Radix, in treating VD. Puerarin can ameliorate cholinergic nervous system disorders, reduce excitotoxicity, anti-inflammation, inhibit apoptosis, alleviate oxidative stress injury, enhance synaptic plasticity, up-regulate neuroprotective factor expression, promote cerebral circulation metabolism, and mitigate Aβ injury. The pathways of action include activating nuclear factor erythroid 2-related factor 2(Nrf2)/antioxidant response element(ARE), vascular endothelial growth factor(VEGF), extracellular regulated protein kinases(ERK), phosphatidylinositol-3-kinase(PI3K)/protein kinase B(Akt), Janus-activating kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3), AMP-activated protein kinase(AMPK), as well as inhibiting the tumor necrosis factor α(TNF-α), transient receptor potential melastatin 2(TRPM2)/N-methyl-D-aspartate receptor(NMDAR), p38 mitogen-activated protein kinase(p38 MAPK), Toll-like receptor 4(TLR4)/nuclear factor-kappaB(NF-κB), early growth response 1(Egr-1), and matrix metalloproteinase 9(MMP-9). By reviewing the papers about the treatment of VD by puerarin published by CNKI, Wanfang, VIP, PubMed, and Web of Science in the last 10 years, this study aims to support the treatment and drug development for VD.

As an emerging pollutant to the environment, microplastics have received widespread attention worldwide. The Loess Plateau, as one of the major agricultural production areas in China, has various land use types, but how the abundance and morphological patterns of microplastics differ among soils under different land use types remains unclear. In this study, we collected soils from three different land use types:croplands, apple orchards, and landfills in the Wangdonggou Catchment. Microplastics were separated and extracted using a modified density centrifugation method, and the abundance, composition, and morphological characteristics of the soil were analyzed and characterized using a laser infrared imaging system. The results showed that the average abundance of microplastics in the Wangdonggou Catchment was 4715 n·kg -1 , mainly composed of PET, PU, and alkyd varnish(ALK), respectively accounting for 30.39%, 29.58%, and 8.42%. More than 80% of the microplastics were fragmented, and more than 60% of the microplastics were of a size ≤ 50 μm. The average abundance of microplastics varied significantly among land use types:cropland soil (7550 n·kg -1 )>apple orchard soil (3440 n·kg -1 )>landfill soil (2283 n·kg -1 ). The average area, width, height, eccentricity, circularity, and other morphological characteristics of microplastics in apple orchard soil were significantly different from those in the cropland and landfill soil.

Chemotherapy is one of the effective methods to treat cancer. However, the chemotherapy agents may cause a series of adverse reactions due to the nonselective characteristics that affect not only tumor cells, but also normal cells. Oral mucositis induced by chemotherapy is a common oral complication caused by chemotherapy in clinic. It brings great suffering to the patients and also interferes with the procedure of chemotherapy. Because of its high incidence in patients receiving chemotherapy and the significant influence, there are more researches on oral mucositis induced by chemotherapy which let us have further understanding of it. This review article will introduce the pathogenesis, risk factors, clinical manifestations, assessments, treatment and prevention of oral mucositis induced by chemotherapy.

Objective: To investigate the clinical manifestations and pathological changes of benign lymphoadenosis of oral mucosa. Methods: The clinical data of 98 cases of benign lymphoadenosis of oral mucosa were analyzed. Results: The clinical manifestations of benign lymphoadenosis of oral mucosa included erosive ulcer (64%) and nodule (9%) and the rate of misdiagnosis was 98%. Neutrophil infiltration occurred in the epithelium of 51% cases and the lymphocyte was diffusely infiltrated in lamina propria of 83% cases. Conclusions: When the mucous membrane of the lamina propria is characterized by complex cell components, diffuse infiltrating lymphocytes and infiltration of neutrophils in mucosal epithelium without erosion and ulceration, it is necessary to highly suspect benign lymphoadenosis of oral mucosa. Finding the focal aggregation of lymphoid follicles or lymphocytes is helpful for the correct diagnosis.

Objective: In this study, the sreening of candidate pathogenic gene is done among family members of an dilated cardiomyopathy(DCM)and hypertrophic cardiomyopathy (HCM) coexistence, and find the relationship between the genotype and the phenotype., Methods: The inheritance atlas was drawn, analysis of genetic characteristics and clinical phenotype.Peripheral venous blood samples of proband and family members were candidated gene exon high-throughput sequencing sub target capture, make the result compares with related database, ultimately screening the target area of the exon and mutations of candidate genes and then using bidirectional sequencing of Sanger to sequence other family members and the health group which were matching with gender and age to testify whether there is the above mutations., Results: In this family, the proband and his father carry three missense mutations, about TTNc.604 A>G(p.Lys202Glu)、TAZ c. 580A>G(p.Ile194Val)and MYH7c.730 T>C(p.Phe244Leu). The heart function of proband was failure, and accompanied malignant arrhythmia.But his father has no obvious clinical symptoms.In this family, the same genetic mutation of disease causing gene lead to different clinical phenotype, but different genetic mutation of disease causing gene lead to the same clinical phenotype.None of the mutations found in this family was found in the health group., Conclusion: The patient of this family carries the genetic mutation of MYH7, TTN and TAZ.The patient of this family carries the composite mutation of MYH7(+) /TTN(+) heterozygous missense mutation and TAZ(+) /TTN(+) heterozygous missense mutation may be show the performance of the genetic characteristics of early onset, severe phenotype.

Objective: To explore the difference between adolescent and adult C57BL/6J mice in response to rapid eye movement sleep (REMS) deprivation in terms of anxiety behavior and hippocampal NO level., Methods: Both adolescent and adult C57BL/6J mice were divided into normal control (NC) group, wide platform (WP) group, and 24-hour REMS deprivation group, each group consisting of 15 mice. REMS deprivation models were established using a small platform in water tank, and the elevated plus maze test was used to examine anxiety behavior of the mice. After behavioral tests, the mice were sacrificed to examine hippocampal NO levels using enzyme-linked immunosorbent assay, and hippocampal nNOS protein expression was detected with Western blotting., Results: The adolescent C57BL/6J mice showed no obvious differences in anxiety behaviors between the 3 groups, but NO level and nNOS expression in the hippocampus was significantly higher in REMSD group than in NC and WP groups (P<0.01). The adult mice in REMSD group, compared with those in the other two groups, exhibited significantly increased total number of arm entry (P<0.01), lowered number of open arm entry and reduced open arm time (P<0.01), increased number of close arm entry and prolonged close arm time (P<0.01 or 0.05); no obvious differences in NO level or nNOS expression in the hippocampus were found in the 3 groups of adult mice., Conclusion: REMS deprivation produces different effects on anxiety-related behaviors between adolescent and adult mice possibly in relation to their different responses in terms of NO levels and nNOS expression in the hippocampus.

To optimize the immunization strategy against HIV-1, a DNA vaccine was combined with a recombinant vaccinia virus (rTV) vaccine and a protein vaccine. Immune responses against HIV-1 were detected in 30 female guinea pigs divided into six groups. Three groups of guinea pigs were primed with HIV-1 DNA vaccine three times, boosted with rTV at week 14, and then boosted with gp140 protein at intervals of 4, 8 or 12 weeks. Simultaneously, the other three groups of animals were primed with rTV vaccine once, and then boosted with gp140 after 4, 8 or 12 weeks. The HIV-1 specific binding antibody and neutralizing antibody, in addition to the relative affinity of these antibodies, were detected at different time points after the final administration of vaccine in each group. The DNA-rTV-gp140 immune regimen induced higher titers and affinity levels of HIV-1 gp120/gp140 antibodies and stronger V1V2-gp70 antibodies than the rTV-gp140 regimen. In the guinea pigs that underwent the DNA-rTV-gp140 regimen, the highest V1V2-gp70 antibody was induced in the 12-week-interval group. However, the avidity of antibodies was improved in the 4-week-interval group. Using the rTV-gp140 immunization strategy, guinea pigs boosted at 8 or 12 weeks after rTV priming elicited stronger humoral responses than those boosted at 4 weeks after priming. In conclusion, this study shows that the immunization strategy of HIV-1 DNA vaccine priming, followed by rTV and protein vaccine boosting, could strengthen the humoral response against HIV-1. Longer intervals were better to induce V1V2-gp70-specific antibodies, while shorter intervals were more beneficial to enhance the avidity of antibodies.

Objective: To investigate the change of lung surfactant protein (SP) A,B,C,D of rats following silica dust exposure in order to provide the evidences for the early diagnosis indices or therapy of silicosis., Methods: 60 male SD rats were randomly divided into silica group, and corresponding controls group. Rats in silica group were administrated 1 ml silica solution by intratracheal instillation at dose of 50 mg/ml. Rats in control group were administrated the same amount saline. At 3rd, 7th, 14th, 21st, 28th after silica exposure, serum and bronchoalveolar lavage fluid (BALF) samples were obtained. The concentration of SP-A, SP-B, SP-C, SP-D in serum and BALF were measured by using enzyme immunoassay (ELISA). Meanwhile the levels of total anti-oxidative activity (T-AOC) and hydroxyproline (HYP) in lung tissue were also detected. The pathology of lung tissue was conducted., Results: Compared with control group, SP-A concentration in BALF of silica exposed rat for 3, 14, 21, 28d was significant lower and SP-D concentration in BALF of silica exposed rat for all time points was also lower. The differences were significant (P < 0.05). Meanwhile SP-B level in 7, 14, 21, 28 d silica exposed rats BALF and SP-C level in 14, 21, 28 d silica exposed rats markedly decreased (P < 0.05). In addition compared with control group, SP-A, SP-B and SP-C concentration in serum of silica exposed rat were higher when SP-A for 14, 21, 28 d silica exposure, SP-B for 7, 14, 21 d silica exposure and Sp-C for 7, 14, 21, 28 d exposure. And all difference were significant (P < 0.05). As silica exposure time increased, SP-C concentration in serum showed an increase trend, which showed a time-response relationship (r = 0.618, P = 0.042). However, SP-D concentration in serum of rat for 7, 14, 21, 28d silica exposure were significant lower than that of control group (P < 0.005). And there was a decrease trend with time point exposure regarding of SP-D (r = -0.731, P = 0.016). The HYP content in lung tissue of experiment rats increased at 3rd, 7th, 14th, 21st and 28th day time point and The T-AOC activity in lung tissue decrease at, 7th, 14th, 21st and 28th day time point. The differences were significant (P < 0.05). There was a positive correlation (P = 0.803, P = 0.045) between SP-C in BALF and HYP of silica exposed rats and a negative correlation between SP-D in BALF and HYP (r = -0.867, P = 0.033). No significant correlation were seen between SP-A, SP-B BALF and HYP (y = 0.416, P = 0.28; r = 0.592, P = 0.071). SP-C concentration in BALF and serum all showed an increased trend and a positive correlation was seen (r = 0.539, P = 0.046). The same decrease trend was seen between SP-D in BALF and serum and correlation value was 0.870 (P = 0.034)., Conclusion: The silica exposure did cause the change of SP content both in BALF and serum. The SP-C and SP-D content in serum might be served as an early effective biomarker of silicosis.

Objective: To explore the effects of acrylamide on the permeability of blood cerebrospinal fluid barrier (BCB) and tight junction protein ZO-1 of choroid plexus in rats and to provide a theoretical basis for explaining the mechanism of nerve injury induced by acrylamide., Methods: Thirty two male Sprague-Dawley rats were randomly divided into ACR and control groups. ACR group was exposed to 20 mg/kg ACR daily for 5 days a week by intraperitoneal injection (i.p.) for 4 weeks. Control group was exposed to normal saline. The neurobehavioral tests (including sensatory and motor functions) were performed every week. At the end of exposure, Evan blue (EB) and Sodium fluorescein (NaFI) content in rat CSF were detected for determining the BCB permeability, Real-time PCR was used to measure the expression levels of ZO-1 mRNA in the epithelium cells of choroid plexus, and laser scanning confocal microscope (LSCM) was utilized to observe the distribution of ZO-1 protein., Results: Neurobehavioral tests showed that the tail-flick latencies of ACR group were 27.77% and 53.71% as long as control group in the 3rd week and 4th week, respectively (P < 0.05). The hind lamb splay distances of ACR group were 131.76% and 153.77% as long as control group in the 3rd week and 4th week, respectively (P < 0.05). Evan blue (EB) and Sodium fluorescein (NaFI) content of ACR group were significantly higher than those of control group (P < 0.05). In the 4th week, the expression level of ZO-1 mRNA in ACR group was 0.21 +/- 0.07, which was significantly lower than that (0.31 +/- 0.11) in control group (P < 0.05). In the 4th week, the ZO-1 protein expression level of choroid plexus in ACR group was significantly lower than that in control group (P < 0.05)., Conclusion: Acrylamide could increased the BCB permeability of rats, which may be involved in the central nervous injury induced by ACR.

In vivo tumor imaging technique method based on bioluminescence principle was established to evaluate the anti-tumor effect of paclitaxel mixed micelle (PMM). MDA-MB-231 tumor cells with luciferase reporter vectors were firstly implanted into nude mice, and subsequently the luciferase substrate was regularly injected during intraperitoneal administration of PMM. Then the tumor size, growth and the intensity of light signals were monitored with in vivo imaging technique. The method of luciferase tumor in vivo imaging could be real-time, reliable and exact in labeling and reflecting the growth of tumors, and the observed results were consistent with that by conventional method, so it would be a feasible approach to study anti-tumor effect of drugs. The anti-tumor effect of paclitaxel mixed micelle was observed by this method, and the results showed that this formulation could inhibit growth of tumor, and the anti-tumor rate of it was about 85%.

Objective: To explore the biomarkers of manganese exposure by measuring the manganese (Mn) and iron (Fe) level as well as the mRNA change of Hepcidin, divalent metal-ion transporter-1 (DMT1) and Parkin-2, one of genes related to Parkinson disease in body fluid and brain tissues of rat., Methods: Male Sprague-Dawley rats were administered (i.p) either MnCl2 solution (6 mg Mn/kg) or the same volume saline, 5 times per week and for 4 weeks. Graphic furnace Atom Absorption Spectrum (AAS) was applied to measure the concentration of Mn and Fe in brain tissue and body fluids. Meanwhile Hepcidin, DMT1 and Parkin-2 mRNA expression were detected by real-time RT-PCR., Results: Mn concentration in erythrocytes of rats was the 86.9 folds of that in control; No significant change was found in plasma. However the trend and range of Mn increase in cerebrospinal fluid (CSF) was the same as that in brain tissue including striatum, cortex, hippocampus and choroid plexus. Meanwhile Fe concentration in brain tissue of Mn exposed rats was also higher than that of control, whose trend was as same as that in CSF. However iron concentration in plasma decreased. The real-time RT-PCR data also showed that Hepcidin mRNA expression in Mn-exposed rat decreased 56% in blood, which was in line with its expression in cortex(67%). Similarly, Parkin-2 mRNA expression decreased both in blood (42%) and in striatum. However DMT1 mRNA expression increase 38% in striatum of Mn-exposed rats but decreased in blood., Conclusion: Hepcidin and Parkin-2 mRNA expression in blood might be serves as the effective biomarkers following manganese exposure, certainly which needs to be further explored.

Aim: To investigated the effect of estrogen on global myocardial ischemia/reperfusion (I/R) injury in ovariectomized (Ovx) rats., Methods: Sprague-Dawley rats were randomly repartitioned into three groups including sham-operated(Sham), ovariectomized (Ovx), or ovariectomized and then given 17beta-estradiol (Ovx + E2). Hearts were excised, mounted on the Langendorff. After the initial stabilization period, all of the three group hearts were randomly divided into normal perfusion subgroup (Control) and I/R perfusion subgroups. Control, perfused for 60 min after stabilization. I/R perfusion subgroups divided into 10 min I + 30 min R, 20 min I + 30 min R, 30 min I + 0 min R, 30 min I + 5 min R, 30 min I + 15 min R and 30 min I + 30 min R. And then, every group hearts were isolated into the single cardiomyocyte. The cardiomyocytes basal contraction and isoproterenol(ISO) stimulation contraction were measured. The viability and yield of cardiomyocytes were counted. LDH and CK concentrations in coronary effluent were assayed with assay kit., Results: The viability and yield of cardiomyocytes were significantly decreased in the conditions of 30 min ischemia followed by different times of reperfusion. The releases of LDH and CK in coronary effluent were significantly increased in the conditions of 30 min ischemia followed by different times of reperfusion. Except the 10 min and 20 min ischemia, the releases of LDH and CK were significantly increased in Ovx during I/R. Ovx + E2 could abate the heart injury through decreasing the releases of LDH and CK. Besides the control and the 10 min I + 30 min R groups, the myocardial basal and ISO stimulation contraction were higher from Ovx than Sham, and the effect was reversed by Ovx + Ez., Conclusion: The results indicate estrogen plays a cardioprotective role in global myocardial ischemia/reperfusion injury in ovariectomized (Ovx) rats.

The accumulation of Enteromorpha prolifera in huge amount in the Yellow sea in June, 2008 draws the attention from all over the world. It is an urgent requirement to monitor the wide range of Enteromorpha prolifera distribution by remote sensing. As to the Enteromorpha prolifera floating on the sea surface, effective monitoring by optical remote sensing has been basically achieved. As far as the underwater suspended Enteromorpha prolifera is concerned, the present paper carried out the radiative transfer simulation research on the above water spectral response, its variation with the suspending depth, the water turbidity and environmental conditions. It was found that with the increase in Enteromorpha prolifera suspending depth and water turbidity as well as the decrease in the thickness of Enteromorpha prolifera, the Enteromorpha prolifera information contained in the surface spectra would decrease. The influence of environmental factors such as water-gas interface roughness, cloud cover extent and sun zenith angle on the underwater suspended Enteromorpha prolifera spectra can be ignored. The maximum Enteromorpha prolifera depth that can cause surface spectrum changes is about 30 m in clean water and about 1 m in turbidity water.

The aim of the present study is to investigate the role of β(2)-adrenoreceptor (β(2)-AR) in ischemic preconditioning (IP) in isolated rat heart model of ischemia/reperfusion (I/R). Sprague-Dawley rat hearts were quickly removed, mounted on Langendorff apparatus, and perfused with Krebs-Henseleit (KH) solution. After the initial stabilization period, the rats were randomly divided into 6 groups including control group (perfused for an additional 20 min), IP group (4 cycles of 5 min of ischemia followed by 5 min of reflow), isoproterenol (ISO) group (10 nmol/L ISO perfusion for 5 min followed by 5 min washout), IP + ICI118551 group (55 nmol/L ICI118551 perfusion for 5 min before and throughout IP), ISO + ICI118551 group (55 nmol/L ICI118551 perfusion for 5 min before and throughout ISO treatment), ICI118551 group (55 nmol/L ICI118551 perfusion for 20 min). After these treatments, all hearts were followed by 30 min of no-flow ischemia and 30 min of reperfusion. A computer-based electrophysiological recorder system was used to measure changes of the maximal rate of pressure increase in systole phase (+dp/dt(max)), maximal rate of pressure decrease in diastole phase (-dp/dt(max)), and difference of left ventricular pressure (ΔLVP). Then cardiomyocytes from these hearts were isolated by 5 min of Ca(2+)-free buffer perfusion and 25 min of collagenase perfusion. The ventricles were chopped and filtered. The myocytes were resuspended in KB buffer. The contraction and the viability of cardiomyocytes were measured. Lactate dehydrogenase (LDH) concentration in coronary effluent was assayed with assay kit. The results showed that both IP and ISO significantly increased the values of ±dp/dt(max), ΔLVP, the contraction and viability of cardiomyocytes, shortened the time-to-peak contraction (TTP), and decreased the release of LDH in coronary effluent. ICI118551, a selective β(2)-AR antagonist, blocked these effects. Either the time-to-50% relaxation (R(50)) or the time-to-100% relaxation (R(100)) had no significant differences between groups. Our results indicate that the cardioprotection of IP was mediated by β(2)-AR in isolated rat hearts subjected to I/R injury.

The preparation and study on the spectra of conductive nano-silver film were done under mild conditions. The silver citrate emulsion coating on the surface of PET was reduced by ascorbic acid (Vc) aqueous solution to prepare a silver film. The morphology, crystal structure, surface roughness and conductivity of the silver film were measured by FTIR, UV-Vis spectrum, X-ray diffraction (XRD), scanning electron microscope, scanning probe microscope system and so on. It was found that the silver citrate emumsion has a homogenous particle distribution from 60 nm-150 nm. The UV-Vis spectrum of the silver film shows a peak at 430 nm, and the silver thin film was considered to have a typical nano-scale structure. By the comparison of the XRD patterns of the films treated and untreated with water, it was concluded that the further deoxidization of silver citrate could be accelerated by water. The crystal structure of the silver film was improved and the surface resistance of the silver film was decreased greatly. The effect of ethanol on the decrease in the resistance of the silver film is much less than that of water.

Fruit of elm has been a popular food in Chinese county for many years. With the rapid development of food nutrition and food safety, more and more people begin to pay attention to its content of trace elements and heavy metals. The wild fruit of elm was studied by ICP-MS/ICP-AES to detect the 22 trace elements. The results showed that the fruit of elm contained many trace elements which are necessary to human health: The concentrations of Mg, Ca, Mn, Fe, Cu, Zn, Sr and Rb were 224.57, 269.73, 9.23, 64.93, 1.68, 4.79, 7.68 and 2.21 microg x g(-1) (FW) respectively; while those of Li, B, V, Co, Ni, Se, Br, Mo and Sn were 318.43, 518.83, 265.52, 108.50, 411.21, 34.51, 51.72, 109.90 and 31.51 ng x g(-1) (FW), respectively. Except the wholesome trace elements, contents of heavy metals (As, Cr, Pb and Cd) are also the important standard to identify the quality of food, and the results showed that the concentrations of heavy metals, Pb, Cd, As, Hg and Cr were respectively 557.87, 8.81, 345.55, 0.78 and 347.97 ng x g(-1) (FW) in fruit of elm, which meet the national hygiene standards.

Objective: To examine the change of Smoothened (Smo) expression in the retinofugal pathway and in the growth cones during the period of embryonic day 13 (E13) to E15., Methods: Smo expression in the chiasm and growth cones was observed by fluorescent immunostaining and retinal explant culture., Results: On E13 and E14, Smo was expressed moderately in the retina and optic disc, and in the corner of the retina, Smo expression was especially dense. On E13, Smo expression was detected in the optic nerves and ventral diencephalon, but only in the superficial region of the optic tract on E14. Smo was also detected in the stem and filopodia of the growth cones in the retinal explant culture during this period., Conclusion: Smo expression changes in different developmental phases, suggesting that Smo might play a role in signal optic axon growth during the development of the retinofugal pathway.

In situ resonance Raman spectra of Chinese freshwater pearls were collected and analysed systematically with three different excitation wavelengths at lambda = 514, 633 and 785 nm. At the same time, the Raman spectra of eggshells of Pomacea canaliculata were also collected under the same experimental conditions in order to compare with pearls' Raman spectra. The conclusions were as follows: (1) Frequency dispersions of Raman spectra of organic pigments were obviously observed in Raman spectra of pearls. In contrast, Raman spectra of organic pigments in eggshells of Pomacea canaliculata showed no frequency dispersion phenomena; (2) It was considered that the organic pigments in pearls were polyacetylenic materials but not carotenoids which was proposed by previous researchers, and the organic pigments in eggshells of Pomacea canaliculata were carotenoids; (3) The conjugated (>=C number of polyacetylenic material in pearls was calculated to be about 10 and 16, and the conjugated C[double bond]C number of carotenoid in eggshells of Pomacea canaliculata was about 13 based on the Raman shifts caused by conjugated C[double bond]C double bonds.

Objective: To express the Gag protein of HIV-1 strain CN54 in Pichia pastoris (P.pastoris), optimize fermentation parameters and purify Gag antigen., Methods: The Gag gene was subcloned into downstream of aox1 promoter of Pichia expression vector pPS1.0, an integrative vector which possesses an identical 5' untranslated region as the natural aox1 gene and employs both in vitro construction and in vivo selection for multi-copy integrants. The recombinant vector was introduced into P.pastoris strain GS115 by electroporation and selected with G418 for Gag gene integration. Super G418 resistant clones were selected and screened for Gag expression. The engineered P.pastoris was cultured to high cell density (>300 A600 Units/ml) in a 5L fermentor. Through methanol induction, the expression level of Gag reached 120 mg/L. Intracellularly expressed Gag was released by high-pressure homogenization and purified through Sepharose FF and DEAE Sepharose FF column chromatography, the purity of Gag reached up to 90%., Results: Western-blotting suggested that purified Gag expressed in P.pastoris could react specifically with serum from HIV infected individual., Conclusion: Gag antigen expressed in P.pastoris has provided a good basis for the development of a new generation of HIV vaccine candidates against some Chinese prevalent strains.

Aim: To prepare the cardiomyocyte-targeting liposomes and investigate their cardiomyocyte targetability in vitro., Methods: Liposomes modified with compound PAC were prepared (PAC-L); The uptake of PAC-L by cardiomyocytes was studied by incubating fluorescence labeled liposomes with cardiomyocytes in vitro and measuring the association of liposomes by a fluorescence spectrophotometer., Results: A high affinity of PAC-L to the cardiomyocytes was observed, the amount of cell uptake of PAC-L by cardiomyocytes was higher than that by nonmyocyte (P < 0.001); The amount of cardiomyocyte uptake of PAC-L on the normoxia condition 2 h was 2.9-fold higher than that of plain-L, and the increase was 5.2-fold when hypoxia occured; The form of liposome uptake changed, the amount of cardiomyocyte uptake of Plain-L by internalization was only 11%, while that of PAC-L was 56%., Conclusion: It is indicated that PAC-L was a potential drug carrier for targeting to ischemic myocardium.

We studied the effect of shear stress (SS) on intercellular adhesion molecule-1 (ICAM-1) expression of rat brain microvascular endothelial cells (RBMECs) using the parallel plate flow chamber method. It was demonstrated that RBMECs showed a time-dependent, but not a force-dependent, upregulation in ICAM-1 expression. Endothelial cell surface expression of ICAM-1 in the supernatants of RBMECs exposed to SS was not changed, thus excluding the possibility that the upregulated expression is due to the factors synthesized by these cells. These data throw light on understanding the signal transduction pathway inside the endothelial cells under the effect of SS.