BACKGROUND: Thrombotic diseases always accompany with blood rheological abnormality. Whether allicin has effects on hemorheological indexes and physiological characteristics of erythrocyte needs to further study.OBJECTIVE: To investigate the effect of allicin on hemorheological indexes and physiological characteristics of erythrocyte in rats with acute blood hyperviscosity.DESIGN: Purely randomly controlled animal study.SETTING: College of Food Science and Nutritional Engineering, China Agricultural University.MATERIALS: A total of 50 Wistar rats of half gender, weighing (200±20) g and aged from 3 to 4 month were provided by Beijing Haidian Tongli Experimental Animal Cultivation Center. Allicin solution was provided by Shanghai Hefeng Pharrnaceutical Co., Ltd. (cencentration: 15 g/L; batch number: 20051001). The main equipments were detailed as follows: NXE-1 cone-plate viscometer (Chengdu Device Factory), DXC-300 nuclear pore membrane erythrocyte deformation radiometer (Device Factory Affiliated to Shanghai Medical University), 650-60 fluorescence polarization (Hitachi Company, Japan) and trace high speed centrifugation (Huaxing Aviation Aircraft Company, Aerospace Faulty).METHODS: The experiment was carried out in the Laboratory of College of Food Science and Nutritional Engineering,China Agricultural University from February to April 2006. All rats were randomly divided into 5 groups, including control group, model group, low-dosage allicin group, moderate-dosage allicin group and high-dosage allicin group, with 10 in each group. Except control group, rats in other 4 groups were perfused with 5 mL tap-water every day fcr 7 successive days. On the seventh day, rats were subcutaneously injected with 0.1% 0.8 mL/kg adnephrin; 2 hours later, rats were dripped in iced water for 5 minutes; 4 hours later, the rats were injected again to establish models of acute blood hyperviscosity. Two days later, all rats in the control group and model group were intraperitoneally injected with saline hematocrit and erythrocyte aggregation index were measured with viscosity determination method; erythrocyte filtration index was measured with nuclear pore membrane flow rate method; erythrocyte membrane fluidity (erythrocyte fluorescence polarization) was measured with fluorescence polarization technique; erythrocyte osmotic fragility test (concentration of NaCl solution from the beginning hemolysis to complete hemolysis) was performed to compare various indexes among different groups.MATN OUTCOME MEASURES: ① Effect of allicin on erythrocyte aggregation (whole blood viscosity,plasma viscosity,hematocrit and erythrocyte aggregation index);② effect of allicin on erythrocyte deformability index (filtration index);③ effect of allicin on erythrocyte osmotic fragility (concentration of NaCl solution from the beginning hemolysis to complete hemolysis);④ effect of allicin on erythrocyte membrane fluidity (erythrocyte membrane fluorescence polarization).RESULTS: Among 50 rats, 8 rats did not have enough blood collection or died; therefore, other 42 rats were involved in the final analysis. ① Effect of allicin on erythrocyte aggregation: As compared with that in the model group, whole blood viscosity was decreased 14.43%, 9.71% and 4.15% in high-dosage, moderate-dosage and low-dosage groups,respectively (P ＜ 0.05-0.01); meanwhile, whole blood viscosity of low shear rate was decreased 15.89%, 14.27% and 6.86%, respectively (P ＜ 0.01), and plasma viscosity was decreased 11.33% (P ＜ 0.05), 9.78% (P ＜ 0.05) and 3.49% (P＞ 0.05), respectively. And the effects mentioned above were shown a dose-effect relationship. Allicin in a high dosage could obviously decrease hematocrit (P ＜ 0.05) and the decreasing degree was 6.47%.② Effect of allicin on erythrocyte deformability index: Filtration index was higher in the model group than that in the control group (0.228±0.025, 0.175±0.043, P ＜ 0.05); however, filtration index was lower in the high-dosage allicin group (0.176±0.034) than that in the model group (P ＜ 0.05). ③ Effect of allicin on erythrocyte osmotic fragility: Concentration of NaCl was higher in the model group than that in the control group at the beginning of hemolysis and at the phase of complete hemolysis. Moreover, allicin in various dosages could improve the increase of erythrocyte osmotic fragility, but there were no significant differences among different dosage groups. ④ Effect of allicin on erythrocyte membrane fluidity: Erythrocyte membrane fluorescence polarization was increased in the model group, and this suggested that membrane fluidity was decreased. Allicin could improve the decrease of erythrocyte membrane fluidity, but there were no significant differences among various dosage groups.CONCLUSION: Allicin can raise erythrocyte deformability, decrease blood viscosity, improve blood fluidity, prevent and cure blood stasis syndrome.

Ligustrazine which has extensive pharmacological effects has been clinically used in the treatment of cardiovascular and cerebrovascular diseases,respiratory diseases,urinary diseases,cirrhosis,cancer,etc.Research of protective effects of Ligustrazine on spinal cord injury has recently increased year by year.Numerous clinical trials and animal experiments suggest that Ligustrazine can play a role on ischemic cerebrovascular disease, degenerative brain disease,epilepsy,spinal cord injury and so on,whose main mechanisms include inhibiting of apoptosis and inflammation,enhancing the body’s antioxidant level,improving hemorheology,increasing expression of neurofilament protein and neurotrophic factors,and regulating balance of the ion intracellular and extracellular.Through consulting the relevant literature since 2003,this article reviews the protective roles and mechanisms of Ligustrazine on cerebra and spinal cord injury for the past 10 years.

In the past, panretinal photocoagulation (PRP) and vitrectomy (PPV) were the main treatments for proliferative diabetic retinopathy (PDR). In recent years, anti-vascular endothelial growth factor (VEGF) drugs have been used more and more widely in PDR due to their advantages in rapidly subtracting new blood vessels, reducing leakage, and promoting the absorption of blood. The combination of anti-VEGF drugs and PRP in the treatment of PDR, especially high-risk PDR, can increase the rate of neovascularization and prevent some patients with mild to moderate vitreous hemorrhage from PPV. The application of anti-VEGF drugs during the perioperative period of PPV can also reduce bleeding during the operation, shorten the operation time, and reduce surgical complications. Although clinical studies have confirmed that anti-VEGF drugs can be used as an alternative treatment for PRP, most patients require multiple and long-term treatments, which increase the psychological and economic burden of patients. It is expected that the cost of anti-VEGF drugs and the development of long-acting dosage forms can be reduced and bring better efficacy and benefits to PDR patients in the future.

Objective To study the inconsistent relation of twin discordancy and the umbilical artery blood flow parameter of systolic/diastolic ratio.Methods 358 pregnant women were divided into the two groups of develop-mental nonidentity( n=67 ) and developmental identity ( n=291 ) according to the criterion that the differences of twins weight exceeded 25 percent.The umbilical artery blood flow parameter of systolic/diastolic ratio were observed. Meanwhile,the difference of weight and the umbilical artery blood flow parameter of systolic/diastolic ratio in the group of developmental nonidentity were observed.Results A significant difference of the umbilical artery blood flow parameter of systolic/diastolic ratio were found in the two groups of developmental nonidentity and developmental identity(7.81%vs 32.56%,P<0.05).There were 56 cases that twins weight exceeded from 25 to 35 percent,7 cases that twins weight exceeded from 35 to 45 percent and 4 cases that twins weight exceeded 45 percent in the group of developmental nonidentity which the umbilical artery blood flow parameter of systolic/diastolic ratio were (23.66 ± 5.78)%,(27.16 ±7.25)% and (33.16 ±4.23)% correspondingly with significant difference(F=23.21,P<0.05 ) .Conclusion The umbilical artery blood flow parameter of systolic/diastolic ratio has significance in diagnosis of twin discordancy.

Objective To prepare icariin-loaded poly(lactic-co-glycolic acid) (PLGA) nanoparticles (icariin nanoparticles) and analyze their physical and chemical properties,the release of icariin,and the effects on human osteoblast cells in vitro.Methods Icariin nanoparticles were prepared by ultrasonic emulsification and dialysis method and characterized by transmission electron microscopy and particle size analyzer.The release studies in vitro were adopted to evaluate the release-control features.Thiazolyl blue tetrazolium bromide (MTT),alkaline phophatase activity assay,and calcium nodes dyeing were carried out to investigate the effects of icariin nanoparticles on the growth and differentiation of human osteoblast cells MG-63.Results The icariin nanoparticles had spherical shapes and uniform particle size,and the mean size of the nanoparticles was (185.8±5.6) nm.The icariin nanoparticles showed the best property of sustained release and biocompatibility.Meanwhile,the icariin nanoparticles had no effect on the growth of MG-63 ceils,but they promote the differentiation of osteoblast cells by a concentration-and time-dependent manner.Conclusions The icariin nanoparticles prepared by ultrasonic emulsification and dialysis method have a good effect on the differentiation of human osteoblast cells in vitro and are biocompatible.

Objective To establish and stabilize a new HLA-B locus typing strategy, reference strand mediated conformation analysis(RSCA)system and to conform its advantages.Methods Of 27 hematopoitic stem cell transplant patients and 63 potential donors DNA samples were extracted from peripheral blood cells and HLA-B loci were both typed by RSCA and PCR-SSP methods. The (ambi)-(guous) results were identified by using DNA sequencing. Results Among 90 samples, 87/90 ((96.7) %) cases could be designated definitely, but one of them was disagreement with the SSP result which was confirmed by sequencing, and 67/90 ((74.4) %) cases could be typed to allelic level. 1/90 ((1.1) %) case could not be identified by RSCA for its bad PCR results. Only one allele of each 2 samples could be designated and the other could not be identified by RSCA, which were further confirmed by sequencing method, and the results confessed which were known as alleles, but there were no corresponding data in RSCA database. Twenty samples were randomly selected to identify the replication rate of RSCA, and the results demonstrated that the replication rate was 100 %. Among PCR-SSP typing results, about 10 % samples might be typed for 1-3 times to confirm their types. Conclusion RSCA had some advantages such as high resolution, high sensitivity, high accuracy, high replication, finding new alleles, large scale and low cost, and was especially suitable for unrelated donor-recipient (screening).

By participating in the national medical colleges and universities students' clinical skill competition and summarizing the experiences after the competition,we reflected on problems and weakness in the past clinical teaching including weak sterile concept and lack of clinical thinking,humanities,communication skills,teamwork awareness,etc.We should take methods to future improve medical students' clinical training capabilities including strengthening the concept of sterile and clinical skills,promoting training of comprehensive clinical thinking ability and problem-solving ability,emphasizing on humane care and communication between doctors and patients; cultivating teamwork awareness thus to comprehensively enhance the overall quality of medical students.

Objective To construct 2 recombinant plasmids carrying cassette chromosome recombinase C(ccrC)and ccrAB respectively and introduce the plasmids into methicillin-resistant Staphlococcus aureus(MRSA)strain 81/0432,and to observe the precise excision of Staphylococcal cassette chromosome mec(SCCmec)island from bacterial chromosome.Methods ccrC and ccrAB genes were amplified with chromosomal DNAs isolated from MRSA strains 81/0342 and N31S as PCR templates.We constructed recombinant plasmids pSR5C and pSR2AB by cloning ccrC and ccrAB genes into temperature-sensitive plasmid pYI3,after introducing them into MRSA strains 81/0432 and N315 by electroporation.PCR was performed to identify SCCmec excision from the bacterial chromosome.The transformants were serial passaged for 10 days,and then the drug resistance of these rransformants was detected by replica experiment.Results The fragment length of ccrC gene was 1.9 kb,smaller than the fragment length of ccrAB from N315.The recombinant plasmids of pSR5C and pSR2AB were successfully constructed.After these 2 recombinant plasmids were introduced into MRSA strain 81/0342,type-V SCCmec island was excised from the chromosome and formed a closed circular structure in the bacteria.However,type-Ⅱ SCCmec island could be excised only in N31S strain after the expression of pSR2AB.Replica experiment verified that transformed strains of 0342(pSR2AB),0342(pSR5C),and N315 (pSR2AB)were mostly susceptible to ceftizoxim or tobramycin after the excision of SCCmec island.Conclusion cciC could serve as a recombinase as ccrAB,which could mediate precise excision of SCCmec island from the chromosome of type-V MRSA strain.This study shows that type-V SCCmec island is widely disseminated between Staphylococcus aweus strains in community setting.

Objective To investigate the in vivo effects of allogeneic mesenchymal stem cells transplantation (MSCT) on OAZ signaling pathway in patients with systemic lupus erythematosus (SLE).Methods Isolated and expand human MSCs from bone marrow cells or umbilical cord of healthy donors were infused into SLE patients. Peripheral blood cells were collected from 10 pre-MSCT patients as well as 1 week and 4 week post-MSCT, and RNA was extracted and reverse transcripted to cDNA. mRNA expression levels of OAZ and Id1-3 were measured by using real-time PCR. Serum levels of IL-10, IL-12, IL-21, CCL2 and ANA were tested by ELISA. Relationships of the gene expression levels with levels of cytokines and ANA were analyzed. Results mRNA expression levels of OAZ, Id1 and Id3 gene in patients with SLE were significantly decreased at week 1(△C:12.4±1.1, 9.7±1.9, 9.7±1.9, 2.1±1.0) and at week 4 (△Ct:13.3±1.2, 10.4±1.5,10.8±1.2, 2.1±1.2) after MSCT when compared to those of the pre-MSCT (△Ct:11.0±0.9, 7.4±2.1, 7.8±2.1, 0.1±1.5 respectively, P all＜0.05). Levels of IL-10, IL-21 and ANA were significantly lower 4 week after MSCT than those before (P＜0.05); while level of CCL2 was higher than pre-MSCT (P＜0.05). Cytokines and ANA levels 1 week after MSCT were not differentially changed comparing to those of the pre-MSCT. Alteration of OAZ mRNA expression levels pre- and post-MSCT were negatively correlated with changes of ANA, IL-21levels and positively correlated with changes of IL-12/IL-10 and CCL2 levels. Conclusions The expression of genes involving in the OAZ signaling pathway is effectively reduced along with the alteration of several cytokines and ANA after allogeneic MSCT in SLE patients. OAZ signaling pathway may play an important role in MSCT treatment for SLE.

Objective To evaluate the protective effect of epigallocatechin gallate (EGCG) against interleukin (IL)-17-induced injury to keratinocytes,and to explore its mechanism.Methods Some cultured HaCaT cells were divided into 3 groups to be treated with IL-17 alone at concentrations of 50,70,90 μg/L,respectively,with those receiving no treatment as the blank control group.Some HaCaT cells were divided into 5 groups:IL-17 group treated with 90 μg/L IL-17 alone,IL-17 + EGCG group treated with 90 μg/L IL-17 and 60 μmol/L EGCG,IL-17 + SP600125 group treated with 90 μg/L IL-17 and SP600125 (a JAK signaling pathway inhibitor),IL-17+ EGCG + anisomycin group treated with 90μg/L IL-17,60xmol/L EGCG and anisomycin (a Janus kinase signaling pathway activator),and blank control group receiving no treatment.After different durations of treatment,CCK-8 assay was performed to evaluate cellular proliferative activity,flow cytometry to detect cell apoptosis,enzyme-linked immunosorbent assay (ELISA) to measure expression levels of IL-6,IL-23 and IL-8,and Western-blot analysis to determine protein expressions of c-Jun N-terminal kinase (JNK) and phosphorylated JNK (P-JNK).Results IL-17 promoted cellular proliferation of HaCaT cells,and the proliferation rate,which was correlated with the concentration of IL-17,reached the maximum in the 90-μg/L IL-17 group (P ＜ 0.05).EGCG at 60 μmol/L significantly inhibited cellular proliferation of,promoted apoptosis in,and reduced IL-6,IL-23 and IL-8 expressions in,HaCaT cells induced by 90 μg/L IL-17 (all P ＜ 0.05).Compared with the IL-17 group,the IL-17 + EGCG group and IL-17 + SP600125 group both showed significantly decreased P-JNK expression,cell proliferation rate and IL-6,IL-23 and IL-8 expression levels (all P ＜ 0.05).However,compared with the IL-17 + EGCG group,the IL-17 + EGCG + anisomycin group showed significantly increased protein expression of P-JNK,cell proliferation rate and IL-6,IL-23 and IL-8 expression levels (all P ＜ 0.05).Conclusion EGCG protected against IL-17-induced injury to HaCaT cells,such as abnormal cell proliferation,apoptosis and inflammatory response,likely by inhibiting the JNK signaling pathway.