Objective To evaluae possible association between interleukin-1 beta (IL-1?) gene exon 5 polymorphism and childhood asthma.Methods The study was conducted in two different groups: asthmatic children(n=55) and healthy children(n=35). The IL-1? gene exon 5 polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP). Results Frequencies of CC,CT and TT genotypes were 92.7%,7.3%,0, and frequencies of C,T allele were 96.4%,3.6% in asthmatic group. However, frequencies of CC,CA and AA genotypes were 85.7%,14.3%,0, and frequencies of C,T allele were 92.9% ,7.1% in healthy group. There were no significant difference in distribution of genotypes and allele frequencies between two groups.Conclusion IL-1? gene exon 5 polymorphism may not be associated with childhood asthma.

Objective To investigate the protective effects of Green Tangerine induced hypertension and mild hypothermia on focal cerebral ischemia and reperfusion and local cerebral glucose utilization(LCGU)in the in- farction rim in rats.Methods A total of 64 rats were used and randomly divided into a control group,a Green Tan- gerine induced hypertension group,a mild hypothermia group and a combination therapy group.The neurologic defi- cits,infarct size and LCGU were observed in the rats with focal cerebral ischemia and reperfusion.Results Com- pared with the control group,the neurologic deficits(P

The stable and controllable quality of decoction pieces is an important factor to ensure the efficacy of clinical medicine. Considering the dilemma that the existing standardization of processing mode cannot effectively eliminate the variability of quality raw ingredients, and ensure the stability between different batches, we first propose a new strategy for Chinese medicine processing technologies that coupled with individuation processed and cybernetics. In order to explain this thinking, an individual study case about different grades aconite is provided. We hope this strategy could better serve for clinical medicine, and promote the inheritance and innovation of Chinese medicine processing skills and theories.
Chemistry, Pharmaceutical ; methods ; standards ; Cybernetics ; standards ; Drug Therapy ; standards ; Drugs, Chinese Herbal ; chemistry ; standards ; toxicity ; Humans ; Quality Control

In this study, we investigated the pharmacokinetics parameters of SPIO-shRNA dual functional molecular probe and observed the main organ distribution by MRI in vivo. Eighteen New Zealand white rabbits were randomly divided into three groups and injected intravenously with different doses of SPIO-shRNA molecular probe, respectively. The blood samples were collected to analyze the pharmacokinetic parameters by measuring the iron content at 30 minutes before and after the injection. Twenty-four Kun Ming (KM) mice were randomly divided into 4 groups: the control group was injected intravenously with physiological saline 200 µL per mouse via the tail vein, the other 3 groups were injected intravenously with different doses of SPIO-shRNA molecular probe. MRI observation was performed in 24 hours, and the liver, spleen, kidney, brain and muscle were collected for iron quantification with Prussian blue staining to determine distribution of the SPIO-shRNA molecular probe in the main organ in vivo. Our results suggest that the molecular probe blood half-life is more than 3 hours. The data of MRI suggest the probe was distributed in liver and spleen, and the MRI signal was reduced with the increase in probe's doses (P < 0.05). The results of Prussian blue staining confirmed the results of MRI. Most of the probe could escape the phagocytosis of mononuclear phagocyte system. Our data provide the pharmacokinetic and distribution of SPIO-shRNA molecular probe in organs. Meanwhile, it suggests the choice of the time and dose of probe for MR imaging of tumor in vivo.
Animals ; Half-Life ; Magnetic Resonance Imaging ; Magnetite Nanoparticles ; Mice ; Molecular Probes ; pharmacokinetics ; RNA, Small Interfering ; chemistry ; Rabbits

Background More and more attentions are paid to the effect of matrix metalloproteinases (MMPs) in the process of wound healing after excimer laser.Objective The present study was to investigate the effects of GM6001,a matrix metalloproteinases inhibitor,on haze formation after excimer laser epithelial keratomileusis (LASIK).Methods LASEK of -10.00 diopter was bilaterally performed on 27 New Zealand white rabbits and then divided randomly into the GM6001 group,Fluorometholone (FML) group and normal control group.Corneal haze was graded under dim light at 2 weeks,4 weeks and 8 weeks after operation,and the number of eyes in different grades of haze were compared among the three groups.Six corneal samples were harvested in each group for confocal microscope examination at 2,4,8 weeks.The corneal histopathological examination was carried out with the optical microscope,and the ultrastructure of the cornea was examined under the transmission electron microscope.Results The numbers of eyes of different corneal grades of haze at 2,4 or 8 weeks after surgery were significantly different among the three groups (P＜0.05).Compared with the normal control group,the corneal haze grades in the GM6001 group and FML group were apparently lower (P＜0.01),but there was no significant difference in the numbers of eyes of different corneal grades of haze between the GM6001 group and FML group (P＞0.01).The numbers of keratocytes in the GM6001 group were apparently lower than those in the normal control group and FML group in 2,4 or 8 weeks after operation (P＜0.05).No significant difference in the keratocytes was seen between the normal control group and FML group at various time points (P＞0.05).Corneal epithelial cell morphology,disordered arrangement of collagen fibers in GM6001 group and in FML group were less pronounced than in the negative control group.Conclusion GM6001 has an inhibitory effect on the formation of corneal haze after LASEK which suppresses proliferation and production of keratocytye with the similar efficacy as FML.

OBJECTIVE Zn-doped CuO nanocomposites (nZn-CuO NPs) are novel nanoparticles synthesized by sonochemical method.This study aimed to further investigate the antitumor effects and mechanism of nZn-CuO NPs, as well as the exact mechanism of reactive oxygen species (ROS) on nZn-CuO NPs-induced death using N-acetylcysteine (NAC). METHODS The antitumor effects of nZn-CuO NPs were evaluated by MTS assay and orthotopic transplantation tumor model in nude mice. The effects of nZn-CuO NPs with or without NAC on ROS production, DNA damage, apoptosis, mitochondrial damage, autophagy, lysosome impairment, and ER and Golgi stress were determined. Also,western blot was used to detect apoptosis and autophagy related proteins,as well as NF-κB pathway related proteins. RESULTS nZn-CuO NPs significantly inhibit tumor growth both in vitro and in vivo. nZn-CuO NPs were able to cause cytotoxicity, ROS production, DAN damage mitochondrial damage, apoptosis, and autophagy, and NAC can attenuate them. Further studies showed that nZn-CuO NPs induced changes of apoptosis, autophagy and NF-κB pathway related proteins, and NAC can restore them. CONCLUSION Overall, our data demonstrated that nZn-CuO NPs could inhibit tumor growth both in vitro and in vivo by ROS-dependent regulation of apoptosis and autophagy, which might be cross-linked by NF-κB pathways.

Objective To explore the effect of complement activation on bone marrow mesenchymal stem cells (BMSCs)and evaluate the effect after transfection of complement regulatory proteins.Methods Bone marrow aspirate was harvested from 10 cases of patients suffered from fractures.Mesenchymal stem ceils were isolated,indentified cultured and then experimented in vitro.The complement cytotoxicity on the mesenchymal stem cells in autologous serum was measured by Europium cytotoxicity assay.The samples were divided into BMSCs group,BMSCs+ autologous human serum (AHS) group and BMSCs+ inactivated autologous human serum (iAHS) group.The complement membrane attack complex (MAC) deposited on the membranes was detected by flow cytometry.Finally,the cytotoxicity on BMSCs was measured after transfected with membrane complement regulatory proteins (mCRPs).All samples were divided into BMSCs with mCRPs untransfected group and BMSCs with mCRPs transfected group.Results More than 95％ of cells derived from bone marrow were identified to be mesenchymal stem cells through detection of cell surface markers by flow cytometry.The cytotoxicity of untreated cells was 0.41％± 1.48％.BMSCs harvested from the 10 patients all had cytotoxicity after incubated with autologous serum,and the cytotoxicity was 32.59％±2.73％,while cytotoxicity after incubated with complement inactivated autologous serum was 2.59％±3.08％,which was similar to control group.Complement attack complex (MAC) could be detected on the BMSCs incubated with autologous serum,which implied the complement activation.After transfection of mCRPs,the cytotoxicity of autologous serum on transfected cells was decreased.The cytotoxicity of untransfected cells (41.70％±4.47％) had significant difference compared to the cells transfected with CD55 (21.87％±2.19％),the cells transfected with CD59 (18.67％± 1.42％),and the cells transfected with CD46+CD55+CD59 (28.43％±2.14％).CD55,CD59 and CD46+CD55 +CD59 transfected groups could impair effectively the cytotoxicity from complement.However,the cytotoxicity impairment was less effective in CD46 transfected cells (39.30％±3.96％),which had no significant difference compared to untransfected cells.Conclusion Membrane complement regulatory proteins could effectively protect bone marrow mesenchymal stem cells from attacks by complement.

OBJECTIVE Cloves(Syzygium aromaticum L.)have been used as both a spice and a traditional Chinese medicinal herb for thousands of years. However, relatively little is known about its potential anticancer activity and mechanisms.In this study,we investigated the in vitro and in vivo anti-tumor effects and mechanisms of water extract of cloves(WEC)against colorectal cancer. METHODS MTS assay and Colony-formation assay were used to detect the anti-tumor activity of WEC on HT-29 cells.The in vivo anti-tumor effect of WEC was detected in a subcutaneous transplantation tumor model of human HT-29 cells.Autophagy was detected by flow cytometry and the expressions of autophagy related proteins(Beclin-1 and LC-3a/b)were determined by western blot. RESULTS MTS result showed that WEC significantly inhibited the viability of HT-29 cells,with the IC50values of 150 μg·mL-1.The colony-formation assay showed that the WEC significantly suppressed colon cancer cells proliferation.WEC also exhibited significant antitumor activity in tumor bearing nude mice. Flow cytometry result showed that WEC significantly induced autophagy, and the averaged relative values of fluorescence intensity were 206,251,341 and 356 in cells treated with 0,100,150 and 200 μg·mL-1WEC for 48 h.Western blot result showed that WEC treatment significantly increased Beclin-1 expression and ratios of LC3-II/LC3-I. CONCLUSION These result showed that WEC inhibited the growth of colon tumor both in vitro and in vivo, which might be related with autophagy induction, and WEC has potential to be developed as a novel anticancer agent for the treatment of colon cancer.

OBJECTIVE Zn-doped CuO nanocomposites (Zn-CuO NPs) are novel nanoparticles synthesized by our research group.In this study,we assessed the in vitro and in vivo antitumor effects of Zn-CuO NPS on pancreatic cancer cells,as well as the potential mechanisms. METHODS MTS assay was used to detect the effects of Zn-CuO NPS on proliferation pancreatic cancer cells(Panc-mia and Aspc-1). The in vivo antitumor effects of Zn-CuO NPs were detected by xenografts model in nude mice. The effects of Zn-CuO NPS on autophagy were detected bytransmission electron microscopy (TEM) andflow cytometry. Autophagy related proteins were detected by Western blotting. RESULTS Zn-CuO NPS significantly inhibited the proliferation of Panc-mia cells and Aspc-1 cells.In vivo experi-ments showed that Zn-CuO NPS significantly inhibited the tumor growth in nude mice without affecting the body weight of the mice. TEM and flow cytometry showed that Zn-CuO NPS induced autophagy, and significantly increased the number of autophagosome.Western Blot showed that Zn-CuO NPS alterd the expression of autophagy related proteins,such as AMPK,mTORand Beclin-1.Also,AMPK inhibitor could significantly reduce Zn-CuO NPS-induced autophagy pathwayas analyzed byWestern blotting. CONCLUSION The findings suggested that Zn-doped CuO nanocomposites inhibited the in vitro and in vivo growth of pancreatic cancer by inducing autophagy through AMPK/mTOR pathway.

OBJECTIVE Granulin A (GRN A), a cytokinesis protein, is derived from proteolysis of progranulin. The previous study in our laboratory has shown that GRN A is able to inhibit cancer cell growth significantly. This study aimed to investigate the effect of combination of GRN A and cisplatin on in vitro and in vivo on the growth of hepatocellular carcinoma. METHODS The in vitro and in vivo antitumor effects of combination of GRN A and Cisplatin were evaluated with MTS assay and subcuta-neous transplantation tumor model.Chou-Talalay method was used to calculate the combination index (CI). Colony formation assay and flow cytometry were used to detect the effects of GRN A on apoptosis. The expression of apoptosis-related proteins were detected by Western blot. RESULTS MTS assay showed that GRN A significantly inhibit hepatocellular carcinoma cells growth with the IC50of 5.6 μmol·L-1, and GRN A combined with cisplatin synergistically inhibit hepatocellular carcinoma proliferation, with the CI<1.The colony-formation assay showed that GRN A significantly enhanced the inhibitory effects of cisplatin on cellular anchorage-independent growth. Flow cytometry showed that GRN A combined with cisplatin synergistically induced apoptosis,with the apoptotic rates of 5.87%,32.74%,35.67% and 67.15% in control, GRN A, Cisplatin, and combination of GRN A and Cisplatin groups, respectively. Western blot confirmed that the two drugs synergistically changed the expressions of proteins related to apoptosis.In vivo experiment indicated that combination of GRN A and cisplatin significantly suppressed tumor growth compared with single drug treatment groups.CONCLUSION The combination of GRN A and cisplatin resulted in synergistic antitumor effects against hepatocellular carcinoma both in vitro and in vivo.