OBJECTIVE:To study the cardiac and skeletal toxicity of retinoic acid (RA) in Danio rerio at early life stage. METHODS:Danio rerio embryos of 24 hours post fertilization(hpf)were used as toxicity model and were exposed under medium with various concentrations of RA(0.1,1,10,25,100 μmol/L). The morphology of embryos and larvae hearts were observed 24,48 h after exposed. LC50 was calculated. Danio rerio larvae of 4 days post fertilization (dpf) were used as skeletal deformity model and were exposed with a series of RA at various concentration(0.1,1,10,25,50μmol/L). They were sacrificed 5 d later, and then Danio rerio skeleton were fixed for staining with alizarin red. The microscopic was used to observe the difference of stained skeleton area. RESULTS:RA caused significant adverse effects on hatching capabilities of Danio rerio embryos,and the ob-vious malformation features were produced during the culture process. 1-100 μmol/L RA could cause heart malformation in Danio rerio embryos and larvae,and the main heart malformation characteristics included heart linearization,pericardial edema,yolksa-cedema,hemocytes accumulation incardiac region. 100 μmol/L RA could inhibit the hatching capabilities of Danio rerio embryos, and caused lethal effects on embryos and larvae. The LC50 were 36.44,23.69 μmol/L after exposed for 24,48 h. 0.1-50 μmol/L RA induced vertebral column sclerotization of Danio rerio embryos and larvae in advance,which was positively associated with the con-centration of RA. CONCLUSIONS:RA can cause cardiac and skeletal toxicity in Danio rerio embryo and larvae,which is positive-ly associated with the concentration of RA.

Zebrafish was selected as model animal, and glucocorticoid dexamethasone was used as a model compound to establish a rapid and high efficient osteopenia model. Zebrafish larvae at 4 days post fertilization (dpf) were exposed to a serial concentrations of dexamethasone solutions, and 0.5% DMSO was selected as the vehicle control group. All groups were incubated in 24-well plates (28.5 degrees C) until 9 dpf. In addition, effects of 10 micromol x L(-1) dexamethasone on preventing against osteopenia induced by etidronate disodium were also investigated. Zebrafish bones at 9 dpf were stained with alizarin red. Quantitative analysis of the stained area was performed by microscopic inspection and digital imaging methods to reflect the amount of bone mineralization. Results showed that dexamethasone group at 2.5, 10 and 25 micromol x L(-1) can decrease the staining area and the staining optical density values of zebrafish head bones when compared with the vehicle control group (0.5% DMSO), which suggested that dexamethasone can significantly reduce the zebrafish mineralized bone and the bone mineral density. Results also showed that 15 and 30 microg x mL(-1) etidronate disodium can increase the mineralized matrix of zebrafish head bone and prevent against osteopenia induced by dexamethasone. In conclusion, the study indicated that zebrafish can be an idea osteopenia model induced by dexamethasone.

Our earlier research has shown that mono-substituted N-phenyl-2, 2-dichloroacetamide exhibited much higher anti-cancer activity than the lead compound sodium dichloroacetate (DCA). In this paper, a variety of multi-substituted N-phenyl-2, 2-dichloroacetamides were synthesized and biologically evaluated. The results showed that 3, 5-disubstituted N-phenyl-2, 2-dichloroacetamide analogues had satisfactory potency. Among them, N-(3, 5-diiodophenyl)-2, 2-dichloroacetamide had an IC50 of 2.84 micromol x L(-1) against non-small cell lung cancer cell line A549 and could induce cancer cell apoptosis.

Model organism zebrafish was used to study metabolism of asperosaponin VI from Dipsacus asper Wall. ex Henry for the first time. Metabolic components of asperosaponin VI after exposing to zebrafish for 24 h were identified by high performance liquid chromatography--electrospray mass spectrometry (HPLC-ESI-MS), the separation was performed with a Zorbax C18 column using a binary gradient elution of 0.05% formic acetonitrile--0.05% formic acid water. The quasi-molecular ions of compounds in both negative and positive mode were observed for molecule mass information, and the potential structures were identified by attentive study on the deglycosylation metabolites and one hydroxylation metabolite of asperosaponin VI. The results were highly in consistent with metabolism of asperosaponin VI in rat. It can be concluded that zebrafish model can wonderfully imitate current metabolic model with advantages of small amount of lower cost, far less amount compound, higher efficiency and more simple, and can reflect integrated metabolism results of in vivo method. Zebrafish metabolic model may become a novel organism model for quick predication on metabolism of even mircoamount compound, which can enrich the available models greatly.

Objective To evaluate the efficacy of probiotics on chemotherapy-induced intestinal mucositis in rats.Methods The databases including PubMed,EMbase,Cochrane,CBM,CNKI,WanFang and VIP were retrieved from their establishment to April 2016.The related randomized controlled trials(RCT) on the effects of probiotics for treating chemotherapy-induced intestinal mocositis in rats were included.The relevant literatures were screened according to the inclusion and exclusion criteria,then the data were extracted and analyzed.Results Total 6 RCT were included.Compared with the control group,the intestinal secretion and absorption function in the probiotics group was strengtnened[SMD=1.73,95 ％CI(0.79,2.68),P＜0.01],jejunal anti-oxidant capacity was increased [SMD=-2.12,95％CI(-3.56,-0.67),P＜0.01],however low dose probiotics (＜1 × 109 cfu/d)had no protective effect on small intestine[SMDjejunum =-0.06,95％CI(-0.51,0.40),SMDileum =0.02,95％ CI(-0.71,0.75);P ＞0.05],while high dose probiotics(≥ 1 × 109 cfu/d) could reduce the intestinal pathological damage[SMDjejunum =-0.64,95 ％ CI (-1.20,-0.09),SMDileum=-0.85,95％ CI(-1.59,-0.12);P＜0.05].Conclusion High dose probiotics can reduce chemotherapy-induced intestinal mucositis in rats.Because of less included literatures and the influence of publication bias,the effect of probiotics on chemotherapy induced mucositis could be overestimated.

Objective To study the correlation between thyroid autoantibodies anti‐thyroid peroxidase antibody (TPOAb) with recurrent miscarriage to seek the objective data indicator for clinical diagnosis of recurrent miscarriage .Methods A total of 1 016 pregnant women of physical examination and normal thyroid function in the obstetric and gynecologic clinic of our hospital from March 2012 to May 2014 were selected as the research subjects .Among them ,90 cases of abortion were screened out and di‐vided into the primary abortion group(60 cases) and the recurrent abortion group(30 cases) .90 healthy childbearing age women of physical examination were selected as the control group .The positive TPOAb cases were performed statistics and compared among various groups ,the ratio was calculated;the TPOAb level was recorded in each group .At the same time the correlation between TPOAb with recurrent abortion was analyzed .Results The TPOAb positive rate in the recurrent abortion group was 46 .67% , which was significantly higher than 25 .00% in the primary abortion group and 4 .44% in the control group;at the same time the TPOAb positive rate of primary abortion group was also significantly higher than that of the control group ,the difference had sta‐tistical significance (P<0 .05) .The TPOAb concentration level in the recurrent abortion group was significantly higher than that in the primary abortion group and the control group;the TPOAb concentration level in the primary abortion group was also signifi‐cantly higher than that in the control group ,the differences were statistically significant (P< 0 .05) .In the follow‐up of adverse pregnancy occurrence with recurrent abortion as the adverse pregnancy event ,and according to the method of Spearman correlation analysis ,with the increase of TPOAb level ,the occurrence rate of recurrent miscarriage was higher ,which showed the positive cor‐relation(r=0 .764 ,P=0 .000) .Conclusion Monitoring the patient′s TPOAb level can better show the symptoms of recurrent abor‐tion .

We cloned human U6 promoter from pAVU6 + 27 vector into pXSN to transcripe small RNA. Meanwhile, a shRNA targeting GDF-8 was cloned down-stream of the hU6 promoter to construct recombinant vector. Then the packing cell GP-293 was co-transfected the recombinant with pVSV-G to gernarate virus particle. Resistant C2C12 cell pools were screened using G418. Levels of mRNA and protein of GDF-8 were tested by Real-Time PCR and western blotting. Cell proliferation and cell cycle were analyzed using MTT and FACS. The expression of GDF-8 was dramatically decreased by the retrovirus-based system in C2C12 cells. Cells proliferated effectively after integrating the recombinant. The cells in G0/G1 phase decreased by 13.7%, while cells in S phase increased by 14.9%. In conclusion, the retrovirus-based RNAi could be used to stably silence GDF-8. It can be a powerful tool in curing muscle atrophy.
Cell Proliferation ; Cells, Cultured ; Down-Regulation ; Gene Expression Regulation ; genetics ; Humans ; Myoblasts ; cytology ; metabolism ; Myostatin ; antagonists & inhibitors ; biosynthesis ; genetics ; RNA Interference ; RNA, Messenger ; analysis ; metabolism ; RNA, Small Interfering ; genetics ; Retroviridae ; genetics ; metabolism

OBJECTIVEThe aim of this study was to investigate the delivery of Cucurbitacin BE to cervical lymph nodes by peri-cancer submucosal injection of the average diameter 85 nm Cucurbitacin BE polylactic acid nanoparticles (CuBE-PLA-NP).

METHODSCervical lymph node metastasis model of mice oral cancer was established and CuBE-PLA-NP and CuBE were injected in peri-cancer submucosal of the mice respectively, the concentration in blood, heart, liver, spleen, lung, kidney and cervical lymph node were measured by high performance liquid chromatography at fifteen time points, Targeted cervical lymph nodes were evaluated by targeting index, selectivity index, targeting efficiency, relative targeting efficiency and ratio of maximum concentration.

RESULTS(1) The CuBE concentration in the cervical lymph nodes after CuBE-PLA-NP injection was far higher than that after CuBE injection at every time point, the CuBE duration in the cervical lymph nodes in CuBE-PLA-NP group was far longer than that in CuBE group; (2) The five targeted values of the cervical lymph nodes in CuBE-PLA-NP group was far higher than that in CuBE group; (3) The CuBE concentration in blood, heart, liver, spleen, lung and kidney in CuBE-PLA-NP group was far lower than that in CuBE group.

CONCLUSIONThe CuBE-PLA-NP can specifically deliver CuBE to the cervical lymph nodes by peri-cancer submucosal injection, increase CuBE concentration and duration in metastasized cervical lymph nodes, and decrease drug concentration of other organs.

Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacokinetics ; Carcinoma, Squamous Cell ; drug therapy ; surgery ; Cucurbitacins ; Delayed-Action Preparations ; Drug Carriers ; Drug Delivery Systems ; Female ; Lactic Acid ; administration & dosage ; pharmacokinetics ; Lymph Nodes ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Mice ; Mouth Neoplasms ; drug therapy ; surgery ; Nanotechnology ; Neck ; Particle Size ; Polyesters ; Polymers ; administration & dosage ; pharmacokinetics ; Triterpenes ; administration & dosage ; pharmacokinetics

OBJECTIVEThe aim of this study was to investigate the target delivery of Cucurbitacin BE (CuBE) to cervical lymph nodes by peri-oral-cancer submucosal injection of the average diameter 85 nm Cucurbitacin BE poly-lactic acid nanoparticles (CuBE-PLA-NP) and evaluate its clinical therapy efficacy.

METHODSCuBE and CuBE-PLA-NP were respectively injected into peri-oral-cancer submucosa at 2, 12, 24, 48, 72, 96, 120, 144, 168 and 192 hours before operation in 26 patients with oral cancer. The concentrations of CuBE in cervical lymph nodes and blood were measured by high performance liquid chromatography (HPLC) at every time point. Cancer cell degeneration and necrosis in metastasis foci of cervical lymph nodes were observed using light microscope and electron microscope.

RESULTS(1) The CuBE concentrations in cervical lymph nodes after CuBE-PLA-NP injection were far higher than those after CuBE injection at every time point, the duration of CuBE existing in the cervical lymph nodes in CuBE-PLA-NP group was far longer than those in CuBE group, the area under the CuBE concentrations-time curve of the cervical lymph nodes in CuBE-PLA-NP group was 43.67 times as many as that in CuBE group; (2) The CuBE ratio of maximum concentrations in the cervical lymph nodes in CuBE-PLA-NP group was 106.46 times as high as that in CuBE group; (3) The CuBE concentrations in the blood in CuBE-PLA-NP group was far lower than that in CuBE group; (4) Cancer cell necrosis and degeneration in the metastasis foci of cervical lymph nodes were found in CuBE-PLA-NP group, necrosis and degeneration were not found in CuBE group.

CONCLUSIONThe peri-oral-cancer submucosa injection of the average diameter of 85 nm CuBE-PLA-NP can specifically delivery CuBE to the cervical lymph nodes, enhance treatment efficiency, and reduce general toxicity.

Adult ; Aged ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacokinetics ; Carcinoma, Squamous Cell ; drug therapy ; surgery ; Chromatography, High Pressure Liquid ; Cucurbitacins ; Delayed-Action Preparations ; Drug Carriers ; Drug Delivery Systems ; Female ; Humans ; Lactic Acid ; administration & dosage ; Lymph Nodes ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Mouth Neoplasms ; drug therapy ; surgery ; Nanotechnology ; Neck ; Particle Size ; Polyesters ; Polymers ; administration & dosage ; Triterpenes ; administration & dosage ; pharmacokinetics

Objective To investigate the expression of immunoglobulin γ-1 heavy chain constant region(IGHG1)in acute myeloid leukemia(AML)THP-1 cells and its influence on cell proliferation,apoptosis,and invasion via its regulation of the transforming growth factor β(TGF-β)/Smad pathway.Methods Bone marrow specimens from nine children with AML and eight children with fractures,human bone marrow stromal HS-5 cells,and human AML THP-1 and HL60 cells were used in the research.Western blot was used to detect IGHG1 protein expression.THP-1 cells were divided into a blank group(without any treatment),si-NC group,si-IGHG1-1 group,si-IGHG1-2 group,si-IGHG1-3 group,TGF-β group,si-IGHG1-1+TGF-β group,and si-IGHG1-1+TGF-β +LY364947 group.Cell proliferation was measured by CCK-8 method.Flow cytometry and Transwell experiment were performed to measure apoptosis and invasion.Western blot was used to detect the protein expression of IGHG1,TGF-β,p-Smad2 and p-Smad3 in each group of cells.Results Compared with the bone marrow of children with fractures,the bone marrow of children with AML(P<0.05)had significantly higher expression levels of IGHG1((0.24±0.03)vs(0.87±0.12)).Compared with HS-5 cells,human AML THP-1 and HL60 cells had significantly increased expression levels of IGHG1((0.89±0.14)(0.75±0.08)vs(0.21±0.02))(P<0.05).Compared with the blank group,the si-IGHG1-1 group of THP-1 cells showed significantly reduced OD450 values(24 h,48 h,72 h),invading cell numbers and protein expression of TGF-β,p-Smad2,p-Smad3 and their apoptosis rate was increased(P<0.05),while the corresponding indexes showed the opposite trend in the TGF-β group(P<0.05).TGF-β reversed the effects of silencing IGHG1 on the proliferation,apoptosis and invasion of THP-1 cells.Compared with the si-IGHG1-1+TGF-β group,the si-IGHG1-1+TGF-β+LY364947 group had significantly decreased TGF-β,p-Smad2,p-Smad3 and protein levels,OD450 values and cell invasion numbers and the apoptosis rate was increased(P<0.05).Conclusions IGHG1 is highly expressed in AML cells.Silencing the IGHG1 gene can inhibit the proliferation and invasion of AML cells and promote the apoptosis of AML cells.This mechanism may be related to the inhibition of the TGF-β/Smad pathway.