OBJECTIVE: To study the tissues distribution of lung targeting Doxorubicin microspheres in mice. METHODS: doxorubicin microspheres were prepared by emulsification-linkage method, and their characteristics was studied. The tissue distribution of doxorubicin microspheres was investigated via caudal vein injection in mice by high performance liquid chromatography. And the targeting character of microspheres was evaluated. RESULTS: Doxorubicin microspheres were orange and regular. Microspheres with diameters of 5 - 15 μm occupied 85.5%, and the loading amount was 4.05%. The re, te and Ce of doxorubicin microspheres was 2.21 times, 1.97 -3.36 times and 1.68 times larger than those of doxorubicin control solution. And the tissue section also indicated that there were more microspheres in the lung tissue. CONCLUSION: Doxorubicin microspheres can target at the lung tissue of miceand reduce the accumulation in heart, which can improve curative effects and reduce toxicity. Copyright 2012 by the Chinese Pharmaceutical Association.

Objective To evaluate the efficiency of primary hepatic carcinoma(PHC)using tran- scatheter arterial chemoembolization(TALE)combining with MSC.Methods Treatment group(30 patients with unresectable PHC)adopt MSC after TALE,while control group(10 patients with unresectable PHC)only adopt TALE,then observing the curative effect and survival rate.Results One year after treating,the local effect was 90 %,60 %,1～2 year survival rate was 90 %,30 % respectively,median life span was 15 months. Conclusion To treat PHC with TALE combining with MSC is an effective and non-invasive method.

Objective To study the antigenicity of human thyrotropin receptor(hTSHR)amino terminus (amino acid 29～280)and its association with Graves' disease.Methods Total thyroid RNA was prepared from human normal thyroid tissue.RNA was then reversely transcripted and cDNA was subjected to PCR amplification.PCR product was cloned into pcDNA3.1 and the recombinant plasmid was named pcDNA3.1/hTSHR188～940bp. Balb/c mice were immunized with peDNA3.1/hTSHR188～940bp. The levels of serum thyroxin,anti-TSHR antibody(TRAb)and thyroid stimulating antibody(TSAb)were measured,and the pathological changes of thyroid tissue were also observed.Results A 753 bp fragment encoding hTSHR ectodomain amino end was obtained after PCR amplification.Confirmed by Hind Ⅲ restriction enzyme digestion and DNA sequencing,pcDNA3.1/hTSHR188～940bphad been constructed successfully,with the correct sequence and direction of hTSHR188～940bp.In the Balb/c mice treated with pcDNA3.1/hTSHR188～940bp,elevated TRAb in week 6(0.148±0.018)were observed compared with those at week o(0.106±0.006,P＜0.01),and kept a higher level till week 10(0.134±0.011,P＜0.01).T4 and TSAb index values were significantly increased in week 10.Serum T4 concentration increased from(41.02±7.97)μg/L in week 0 to(62.20±12.77)μg/L in week 10(P＜0.01);TSAb index values rose from 0.864±0.076 at week 0 to 1.392±0.615(P＜0.01).Thyroid pathological examination showed that proliferated thyroid follicular epithelial cells and foll icular eapacity increased.Inflammatory cells were occasionally found.Conclusions There are antigen epitopes in hTSHR ectodomain amino acid 29～280,which can stimulate the production of TSAb.And the latter induces hyperthyroidism and Graves' disease like manifestations.It suggests that hTSHR ectodomain amino acid 29～280 is closely associated with Graves' disease,and maybe one of important etiological factors leading to the disease.

Objective To study the current situation of human brucellosis infection in a population at high risk in Qian'an,and to provide a scientific basis for prevention and control of the disease.Methods Towns with centralized residents working in sheep breeding,transporting,slaughtering and processing in Jianchangying,Muchangkou and Xiaguanying of Qian'an were selected.In each selected town,2-3 villages with relatively centralized households working in sheep farming,transportation and slaughtering were chosen.All of the people who contacted the sheep or their excrement were chosen as monitoring objects,and serological antibody was tested with rose Bengal plate test(RBPT) and serum agglutination test(SAT).Regional,gender,age and occupational distribution of brucellosis were analyzed.Results A total of 367 blood samples were tested,46 of them were positive in both RBPT and SAT with a ratio of 12.53％ (46/367).Male positive rate [13.51％ (30/222)] was slightly higher than that of females [11.03％(16/145)].The rate in Jianchangying was higher than that of other two towns with a ratio of 13.38％(40/299).The veterinary population had the highest ratio of 33.33％(1/3).Conclusions It is necessary to carry out the surveillance on brucellosis and to further strengthen communication with the animal husbandry department,and strengthen protection on key population.At the same time,in order to control the spread of the disease,extensive health education and intervention measures should be carried out.

It was difficult to prepare traditional Chinese medicine pellets due to the adverse characteristics of the herbal extract. In this study, Danshen extract (DS) powder mixed with different proportions of microcrystalline cellulose (MCC), lactose and starch were made into pellets by extrusion-spheronization. Particle size, span, bulk density, tapping density, compressibility, Hausner ratio and angle of repose were used to evaluate the micromeritic properties of mixing powders. Feret diameter, aspect ratio, yield, density and friability were used to evaluate the properties of the pellets. The correlations between micromeritic properties of raw material powders and the formability of their pellets were analyzed by cluster analysis, principal component analysis and partial least squares regression analysis. As a result, the particle size of the powders was negatively correlated with the size, density, yield, and was positively correlated with the friability of their pellets. The span, density, compressibility and angle of repose of the powders were positively correlated with the size, density, yield, and were negatively correlated with the friability of their pellets. So there were certain correlations between the micromeritic properties of raw material powders and the properties of their pellets prepared by extrusion-spheronization. This research provided a foundation for the technology and method of traditional Chinese medicine extract pellets.
Cellulose ; chemistry ; Drug Compounding ; methods ; Drug Implants ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Excipients ; chemistry ; Lactose ; chemistry ; Medicine, Chinese Traditional ; methods ; Particle Size ; Powders ; chemistry ; Starch ; chemistry

Air Pollutants, Occupational ; analysis ; Female ; Hexanes ; analysis ; poisoning ; Humans ; Male ; Occupational Exposure ; statistics & numerical data ; Shoes

OBJECTIVETo investigate the chemical constituents of Fraxinus paxiana.

METHODThe chemical constituents were isolated and purified by chromatographic techniques and the structures of the compounds were identified with or by spectroscopic methods.

RESULTFifteen compounds were obtained from the methanol extract of F. paxiana and their structures were elucidated as esculin (1), esculetin (2), fraxin (3), fraxetin (4), salidroside (5), osmanthuside H (6), liriodendrin (7), 3-(4-beta-D-glucopyranosyloxy-3-methoxy)-phenyl-2E-propenol (8), threo-syringylglycerol (9), euscaphic acid (10), 3-hydroxy-1-(4-hydroxy-3, 5-dimethoxyphenyl)-1-propanone (11), omega-hydroxypropioguaiacone (12), sinapyladehyde (13), betulinic acid (14) and mannitol (15).

CONCLUSIONAll compounds were obtained from this plant for the first time.

Coumarins ; chemistry ; Esculin ; chemistry ; Fraxinus ; chemistry ; Furans ; chemistry ; Glucosides ; chemistry ; Glycosides ; chemistry ; Mannitol ; chemistry ; Methanol ; chemistry ; Phenols ; chemistry ; Plant Bark ; chemistry ; Triterpenes ; chemistry ; Umbelliferones ; chemistry

Background Although Leber hereditary optic neuropathy (LHON) and optic neuritis have different causes and managements,their clinical manifestations are difficult to be distinguished.Real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR) is a high flux,simple,rapid and specific detecting technology,so establishing a specific diagnosis method of LHON with RTFQ-PCR has a practical significance.Objective Purpose of the present study was to establish a real-time Taqman probe for mitochondrial DNA (mtDNA)11778G＞A mutation in LHON patients.Methods Primers and Taqman probe for mtDNA 11778G＞A mutation were designed based on mtDNA complete geneme.Eighty-four patients with LHON were selected from the LHON DNA bank of Molecular Biology Laboratory,Henan Eye Institute,and 40 normal physical examinees aged 18-20 years were from Henan People's Hospital.2 ml of periphery blood was collected from each individual.Based on the double-blindness principle,mtDNA 11778G＞A mutation was tested by both Taqman probe and sequencing to check the reliability of real-time Taqman probe.Results The mtDNA 11778G＞A mutation was found in 23 out of 84 patients,and 61 showed a negative result by the technique of real-time Taqman probe.The Ct values of 23 patients with mtDNA 11778G＞A mutation were 22.993 ±0.708,but those of 5 normal controls were 0.These findings showed a consistent rate of 100％ with the sequencing results.In addition,both the false positive rate and the false negative rate were zero.Conclusions Real-time Taqman probe technique is an accurate,convenient,sensitive,specific and intuitionistic method for the diagnosis of mtDNA 11778G＞A mutation in LHON patients.It is feasible and suitable to screen the LHON patients with mtDNA 11778G＞A mutation in a large scale.

OBJECTIVETo observe the influence of lead exposure on the activity of nitric oxide synthase (NOS) in different brain regions of rat.

METHODSBy establishing a series of rat models exposed to different low levels of lead (drinking water containing 0.025%, 0.050%, 0.075% of lead acetate) during developing period, NOS activities in hippocampus, cerebellum, cerebral cortex and brain stem were studied.

RESULTSOn the 21st day after birth, NOS activities in hippocampus of three levels of lead exposed groups [(1.53 +/- 0.20), (1.66 +/- 0.23), (1.88 +/- 0.32) U/mg pro respectively], and in cerebellum [(0.87 +/- 0.24), (0.85 +/- 0.09), (0.91 +/- 0.18) U/mg pro respectively] were significantly lower than those of control group [(2.36 +/- 0.18), (1.41 +/- 0.18) U/mg pro, respectively, P < 0.01]. NOS activities in cerebral cortex of 0.075% group [at 7, 14, 21 d of age [(1.29 +/- 0.14), (1.03 +/- 0.15), (0.69 +/- 0.10) U/mg pro] were significantly lower than those in control group [(2.54 +/- 0.31), (1.64 +/- 0.22), (1.24 +/- 0.14) U/mg pro respectively], and 0.025% group [(2.42 +/- 0.19), (1.59 +/- 0.17), (1.27 +/- 0.12) U/mg pro respectively], and 0.050% group [(2.56 +/- 0.53), (1.77 +/- 0.19), (1.24 +/- 0.10) U/mg pro respectively, P < 0.05]. There were no significant differences among control, 0.025%, and 0.050% groups (P > 0.05). Lead exposure had no influence on NOS activity in brain stem at the same age (P > 0.05).

CONCLUSIONNOS activities in hippocampus, cerebellum and cerebral cortex were inhibited by low level lead exposure and the degree of the effect was related to Pb exposure time and/or level of Pb exposed.

Animals ; Brain ; drug effects ; enzymology ; Brain Stem ; drug effects ; enzymology ; Cerebellum ; drug effects ; enzymology ; Cerebral Cortex ; drug effects ; enzymology ; Dose-Response Relationship, Drug ; Female ; Hippocampus ; drug effects ; enzymology ; Lead ; toxicity ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Time Factors

OBJECTIVETo study the roles of stromal interaction molecule 2 (STIM2) and transient receptor potential canonical 3 (TRPC3) in extracellular Ca(2+)-sensing receptor (CaR)-induced extracellular Ca2+ influx and the production of nitric oxide (NO) in human umbilical vein endothelial cells (HUVEC).

METHODS(1) The interaction of STIM2 and TRPC3 was determined using the immunofluorescence technique. (2) The expressions of STIM2 and TRPC3 genes were silenced in HUVEC by transfection constructed STIM2 and TRPC3 RNA interference plasmids. The interference efficiency of STIM2, TRPC3 protein and mRNA levels were determined by Western blot and real time RT-PCR, respectively. (3) The second to fifth passage of HUVEC were divided into: STIM2-002 short hairpin RNA (STIM2-002 shRNA ) + spermine + Ca2+ group and TRPC3-004 short hairpin RNA (TRPC3-004 shRNA ) + spermine + Ca2+ group; control group (spermine + Ca2+ group) and vehicle+ spermine + Ca2+ group. The four groups of cells were incubated with CaR agonist spermine, the intracellular Ca2+ concentration ([Ca2+]i) was detected using the fluorescence Ca2+ indicator Fura-2/AM, and the production of NO was determined by DAF-FM (NO fluorescent probe) of each group in HUVEC.

RESULTS(1) Immunofluorescence technique results showed that STIM2 and TRPC3 proteinswere present in the cytoplasm of HUVEC. (2) The results of transfection constructed STIM2 and TRPC3 RNA interference plasmids demonstrated that shRNA targeted to the STIM2 and TRPC3 genes decreased STIM2 and TRPC3 mRNA levels by 88.2% and 74.0%, respectively (P < 0.05), simultaneously, the STIM2 and TRPC3 protein levels were decreased by 79.9% and 71.8%, respectively (P < 0.05). (3) Compared with spermine + Ca2+ group, the [Ca2+]i and the net NO fluorescence intensity of spermine + Ca(2+) + ShSTIM2-002 group, spermine + Ca(2+) + ShTRPC3-004 group and spermine + Ca2+ Vehicle group were not changed (P > 0.05).

CONCLUSIONSTIM2 and TRPC3 do not participate in CaR-mediated Ca2+ influx and NO production individually.

Calcium ; metabolism ; Cell Adhesion Molecules ; physiology ; Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; physiology ; Humans ; Nitric Oxide ; metabolism ; Stromal Interaction Molecule 2 ; TRPC Cation Channels ; physiology