Objective:This study was performed using preclinical transplanted animal experiments to analyce the effects and mechanisms of third-generation EGFR-TKIs combined with anti-angiogenic therapy, thereby providing theoretical basis for further clinical trials. Methods:Researchers constructed the transplant BALB/C nude mice models with H1975 lung adenocarcinoma cell line (EGFR T790M) and divided the mice into four groups and treated them with osimertinib (2.5 or 5 mg/kg/day, gavage) alone or plus bevacizumab (5 mg/kg/twice weekly, i.p.) when the tumors reached approximately 0.4-0.6 cm3 in volume. The tumor growth curve of tumor volume was drawn according to the time in every group. After 2 weeks of treatment, the mice were killed and the tumors were processed for immunohistochemical staining and Western blot analysis. Immunostaining was performed to detect:HIF-1α, VEGF, and microvessel density (MVD) by using SP method on paraffin sections. Western blot analysis was used to analyze the protein expression levels of EGFR, AKT, and ERK signal transduction pathways. Results:After 2 weeks of treatment in high-and low-dose osimertinib alone, tumor volume in the high-dose group was significantly less than in low-dose osimertinib-alone group (P<0.05). VEGF, HIF-1αexpression, and MVD were significantly low in the high-dose osimertinib-alone group (P<0.05). Increasing doses of osimertinib induced dose-dependent weakening of the p-EGFR, p-AKT, and p-ERK expression levels (P<0.05). In the low-dose osimertinib-plus-bevacizumab group and low-dose osimertinib-alone group, no significant difference in tumor volume and the above factors was observed. In the low-dose osimertinib-plus-bevacizumab group and high-dose osimertinib-alone group, tumor volumes did not exhibit significant difference (P=0.178). Moreover, VEGF, HIF-1αexpression, and MVD exhibited no significant difference. No significant difference in the p-EGFR, p-AKT, and p-ERK expression levels was found between high-dose osimertinib-alone group and low-dose osimertinib-plus-bevacizumab group (P>0.05). In the high-dose osimertinib-plus-bevacizumab group, tumor growth was not significantly greater than that in the high-dose osimertinib-alone group (P=0.642). No significant difference was observed in the above factors.In the high-and low-dose osimertinib-plus-bevacizumab groups, tumor volume and the above factors did not exhibit significant differences (P>0.05). Conclusion:Osimertinib has obvious antitumor effects in EGFR-mutant lung adenocarcinoma with T790M mutation cell xenografts. Bevacizumab has a synergetic inhibitory effect with osimertinib against EGFR-mutant lung adenocarcinoma with T790M mutation cell xenografts. Bevacizumab enhanced the antitumor effects of osimertinib by reducing VEGF expression and the microvascular density of the tumor, thereby improving the tumor microenvironment. Bevacizumab can enhance the effect of osimertinib by suppressing EGFR, ERK, and AKT phosphorylation, thereby synergistically inhibiting EGFR activation and downstream signaling.

Objective To summarize the clinical features of mercury poisoning diagnosed by blood and urine tests for improving the diagnosis and treatment of the disease.Methods Poisoning causes,clinical manifestations,diagnosis,treatment and prognosis were retrospectively reviewed in 92 in-patients with mercury poisoning in our hospital from January 2000 to April 2010.Results Of the 92 patients,37 were male and 55 were female with an average age of 33.1(2-65)years old.The mercury poisoning was caused by occupational exposure and non-occupational exposure,such as iatrogenic exposure,life exposure and wrong intake or suicidal intake of mercury-containing substances,mainly through respiratory tract,digestive tract and skin absorption.The most common clinical symptoms were as the followings:nervous system symptom,such as memory loss in 50 eases(54.3%),fatigue in 34(37.0%),numb limb in 25 (27.2%),dizziness and headache in 22(23.9%),cacesthesia in 20(21.7%),fine tremor(finger tip,tongue tip,eyelids)in 15(16.3%),insomnia and more dreams in 12(13.0%);gastrointestinal symptoms:nausea in 16 (17.4%),abdominal pain in 14(15.2%),stomatitis in 5(5.4%);joint and muscle symptoms:muscle pain in 16(17.4%),joint pain in 5(5.4%);cardiovaseular system:chest tightness,hean palpitations in 6(6.5%);urinary system:edema in 9(9.8%);other system:hidrosis in 20(21.7%).After the treatment with sodium dimercaptopropane sulfonate (DMPS),the symptoms were gradually alleviated.Their gastrointestinal,cardiovascular symptoms were alleviated within 2 weeks;neurological symptoms were alleviated within 3 months;kidney damage showed a slower recovery and could be completely'alleviated within 6 months.Conclusions Because of its diverse clinical symptoms,the mercury poisoning was easy to misdiagnosis and missed diagnosis:therefore the awareness of the disease should be further enhanced.Leaving from the poisoning environment timely and giving appropriate treatment with DMPS will lead to a satisfactory prognosis.

Carcinoma ; genetics ; metabolism ; pathology ; surgery ; China ; Humans ; Male ; Oncogene Proteins, Fusion ; genetics ; Prostatic Hyperplasia ; genetics ; metabolism ; pathology ; surgery ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Proto-Oncogene Proteins c-ets ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Serine Endopeptidases ; genetics ; metabolism

Objective To investigate the expression of microRNA-21(miRNA-21)in two human malignant melanoma cell lines,A375 and M14,and its effect on the viability of these cells.Methods The expression of human miRNA-21 was assessed by quantitative fluorescent PCR in A375 and M14 cells.Then,three concentrations(90,180,270 nmol/L)of miRNA-21 inhibitor and were transfected both cells and a negative control were transfected a mixture of LipofectamineTM 2000 reagent,respectively.After another 3-day culture,the proliferation of cells was detected by cell counting kit-8,and R value was calculated to denote the relative activity of cells.Statistical analysis was carried out by SPSS13.0.Results The expression of miRNA-21 was higher on A375 cells than that on M14 cells with the average value of 2-deltaCT being 1.2928±0.1509 vs 0.1894±0.1803.With miRNA-21 inhibitor at the concentration of 90,180,270 nmol/L,the activity of A375 cells was significantly lowered in comparison with that in the control group,with the R value being 0.7362±0.1662.0.7248±0.3204 and 0.6767±0.2998 respectively(all P＜0.01).However,in the case of M14 cells,cell activity was only suppressed by miRNA-21 inhibitor at 90 nmol/L with the R value being 0.7295±0.1478.and no significant inhibition was observed with the inhibitor at 180 or 270 nmol/L (both P＞0.05).Conclusions miRNA-21 is expressed on human melanoma cell lines,A375 and M14,at different levels,with a promoting effect on the proliferation of both cells.Moreover,miRNA-21 may act as an oncogene-like gene via down-regulating the expression of some tumor-inhibiting factors.

Objective To evaluate the imaging performance and characteristics of peripheral primitive neuroectodermal tumors (pPNET), in order to raise awareness of the disease and the diagnostic accuracy. Methods Thirteen patients with pPNET who confirmed by puncture or pathology were enrolled in this study. The CT and MRI features were retrospectively analyzed. Four patients received simple routine CT examination, and 9 patients had MRI examination based on X-ray, CT examination. Results The part of body involved was more widely.Two patients occurred in the sinuses, 1 patient in the chest wall, 1 patient in the lung, and 3 patients in the abdominal and pelvic(1 patient involved the pelvic floor through the pelvic wall lesions to basins), 1 patient in the spine, and 5 patients located in the extremities. The characteristics of pPNET by CT and MRI in soft tissue of 8 patients showed single large soft tissue mass, and the biggest was 11.2 cm × 10.6 cm. In bonds and joints of 5 patients, CT and MRI showed wide range of soft tissue mass, mixed internal signal/density with obviously uneven enhancement, significant bone destruction and dissolvable osseous changes, and no obvious tumor bone and calcification. Conclusions Each site has its relative imaging characteristics of pPNET. CT and MRI can show lesions involving the scope, internal structure, with or without adjacent tissue invasion and metastasis, and provide guidance for clinical treatment and selecting treatment regimen. But in the end, it depends on the diagnosis of pathological and immunohistochemical examination.

BACKGROUND:The carriers made from biodegradable and biocompatible polymeric materials represent an exciting approach to increase the bioavailability and stability of oral y administered insulin by the chemical reaction or physical encapsulation of insulin.
OBJECTIVE:To mainly review the research progress of the material types, preparation methods, physicochemical characteristics, in vitro release kinetics, and bioavailability of polymeric materials adopted as oral insulin carriers.
METHODS:The first author searched PubMed, Elsevier and CNKI databases for articles (2002-01/2013-02) concerning the polymeric materials and oral insulin carriers with the key words of“polymeric biomaterials, oral insulin, carrier”in English and in Chinese.
RESULTS AND CONCLUSION:Currently, there are mainly two kinds of polymeric biomaterials used for oral insulin delivery systems, that is, natural polymeric biomaterials (such as chitosan and alginates) and synthetic polymeric biomaterials. The most commonly used synthetic polymeric materials for the preparation of these vehicles are polyesters, polyacrylates and their copolymers, which are wel known for their good biodegradability, biocompatibility, and physiological properties. Although researchers have tried to develop promptly oral insulin formulation using various technologies, the reports about clinical application or commercial success have not been seen because of several questions such as polymer material as a carrier, the lower bioavailability of insulin, the quality standards and stability of the formulation. Hence, future studies wil focus on the development of a new type of polymer-based material as carriers by choosing the new materials or modifying physical and chemical characteristics of existing polymers, to avoid gastrointestinal destruction of the insulin and increase bioavailability of insulin in the body, so as to obtain the good control ed release rate and effect.

Objective To observe the clinical effect of conservative therapy on prolapse of lumbar intervertebral disc(PLID).Methods 365 PID patients included 78 acute cases and 287 subacute cases.The treating principle for acute cases was eliminating edema and aseptic inflammation,and that for subacute cases was releasing adhesion and removing inflammatory stimulation to nerve root.Results Of 78 acute patients,60 cases(76.9%) healed,18 cases(23.1%) improved;of 287 subacute patients,186 cases(64.8%) healed,101 cases(35.2%) improved,the effective rate of 365 patients was 99.7%.Conclusion Conservative therapy is effect on PLID at acute and subacute periods.

OBJECTIVE: A systematic study of Laggera pterodonta was performed in order to identify its bioactive compounds. METHODS: Solvent partition and column chromatography used to isolate and purify Laggera pterodonta compounds. The isolated compounds were further elucidated by high field NMR spectroscopy. RESULTS: Fifty-two compounds were separated and the structures of 39 compounds were elucidated. Twenty-four compounds have not been previously reported. For example, the structure of compound 19, a previously undetected compound was elucidated as 4gamma, 9alpha,11-triol-enantio-eudesmane using both 1D and 2D NMR spectroscopy. Major secondary metabolites of the Laggera pterodonta included eudesmane-type sesquiterpenes and flavone derivatives. CONCLUSION: A systematic study of a yunnan herbal medicine Laggera pterodonta resvealed several novel compounds that may have clinical significance. Further in vivo animal studies should be performed to assess their bioactive role.

An in vitro P-glycoprotein mediated drug biliary excretion model (B-Clear model) was developed and validated using sandwich-cultured rat hepatocytes (SCRH) and a model substrate rhodamine 123 (Rh123). SCRH formed functional bile canalicular networks after 5 days of culture. Rh123 (10 micromol x L(-1)) was then incubated with the SCRH in standard Ca+ Hanks buffer or Ca(2+)-free buffer. The cumulative cell uptake and canalicular efflux of Rh123 under Ca2+ and Ca(2+)-free conditions were measured with a LC-MS/MS method. The biliary excretion index (BEI) and instinct biliary clearance (CL(bile, int)) were calculated. To assess the effect of known P-gp inhibitors on the efflux of Rh123, cyclosporine A (CyA), tariquidar (TQD) or quinidine (QND) (10, 50 and 100 micromol x L(-1)) was pre-incubated separately with SCRH for 30 min, then co-incubated with Rh123. The BEI and CL(bile, int) of Rh123 obtained from the SCRH model were (17.8 +/- 1.3) % and (10.7 +/- 0.9) mL x min(-1) x kg(-1), respectively. All the three P-gp inhibitors showed a dose-dependent inhibition on the bile clearance of Rh123, indicating that the B-Clear model with SCRH was functional properly. The biliary excretion of loperamide (LPAD) and the role of P-gp were further investigated with this validated model. The BEI and CL(bile, int) for LPAD (20 micromol x L(-1)) were obtained after it was incubated with SCRH for 30 min, and found to be (12.9 +/- 1.2)% and (6.1 +/- 0.3) mL x min(-1) x kg(-1) respectively. The dose-dependent inhibition on LPAD biliary excretion by CyA, TQD or QND confirmed the major role of P-gp in LPAD canalicular efflux. The results suggested that the B-Clear model with SCRH would be a useful tool for evaluation of P-gp mediated efflux and drug-drug interaction.
ATP-Binding Cassette, Sub-Family B, Member 1 ; antagonists & inhibitors ; Animals ; Biliary Tract ; metabolism ; Cells, Cultured ; Chromatography, High Pressure Liquid ; Cyclosporine ; pharmacology ; Hepatocytes ; cytology ; metabolism ; Loperamide ; metabolism ; Male ; Quinidine ; pharmacology ; Quinolines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Rhodamine 123 ; metabolism ; Tandem Mass Spectrometry

OBJECTIVETo investigate the effect of transfusion of necrotic cells on regulatory T (Treg) and Th17 cell balance in septic mice.

METHODSThirty-four C57BL/6 mice were randomized into PBS group (n=5), sham-operated group (n=5), sepsis group (n=12), and necrotic cell transfusion group (n=12) and subjected to intraperitoneal PBS injection, sham operation by separating the cecum only, cecal ligation and puncture (CLP), and injection of 2 × 10⁷ necrotic cells 5 days before CLP, respectively. All the mice were sacrificed 2 weeks after CLP for analyzing the proportion of CD4⁺Foxp3⁺Treg cells and CD4⁺IL17A⁺Th17 cells in the peripheral blood, spleen and thymus by flow cytometry.

RESULTSThe percentage of Th17 cells and Treg/Th17 ratio in the spleen was significantly higher in CLP group than in the sham-operated group and PBS group (P<0.01). The percentage of Treg cells in the thymus was significantly lower in CLP group than in the sham-operated group (P<0.01). Pre-infusion of necrotic cells redressed the abnormality of Treg and Th17 cell percentages and Treg/Th17 imbalance in mice following CLP (P<0.05).

CONCLUSIONPre-infusion of necrotic cells can reverse Treg/Th17 imbalance in septic mice.

Animals ; Cecum ; Flow Cytometry ; Interleukin-17 ; Ligation ; Mice ; Mice, Inbred C57BL ; Necrosis ; Sepsis ; therapy ; Spleen ; T-Lymphocytes, Regulatory ; cytology ; Th17 Cells ; cytology ; Thymus Gland