The aimed of this study was to prepare stabilized thiomers to overcome the poor stability character of traditional thiomers. Poly(acrylic acid)-cysteine (PAA-Cys) was synthesized by conjugating cysteine with poly(acrylic acid) and poly(acrylic acid)-cysteine-6-mercaptonicotinic acid (PAA-Cys-6MNA, stabilized thiomers) was synthesized by grafting a protecting group 6-mercaptonicotinic acid (6MNA) with PAA-Cys. The free thiol of PAA-Cys was determined by Ellmann's reagent method and the ratio of 6MNA coupled was determined by glutathione reduction method. The study of permeation enhancement and stabilized function was conducted by using Franz diffusion cell method, with fluorescein isothiocyanate dextran (FD4) used as model drug. The influence of polymers on tight junctions of Caco-2 cell monolayer was detected with laser scanning confocal fluorescence microscope. The results indicated that both PAA-Cys and PAA-Cys-6MNA could promote the permeation of FD4 across excised rat intestine, and the permeation function of PAA-Cys-6MNA was not influence by the pH of the storage environment and the oxidation of air after the protecting group 6MNA was grafted. The distribution of tight junction protein of Caco-2 cell monolayer F-actin was influenced after incubation with PAA-Cys and PAA-Cys-6MNA. In conclusion, stabilized thiomers (PAA-Cys-6MNA) maintained the permeation function compared with the traditional thiomers (PAA-Cys) and its stability was improved. The mechanism of the permeation enhancement function of the polymers might be related to their influence on tight junction relating proteins of cells.

The transcellular process of coumarin 6 (C6) loaded poly(ethyl ethylene phosphate)-co-poly (epsilon-caprolactone) (PEG-PCL) micelles on Madin-Darby Canine Kidney (MDCK) epithelial cells was investigated. C6 loaded PEG-PCL micelles were prepared using the thin-film hydration method. The size of the micelles was characterized by dynamic light scattering analysis using a Malvern Zetasizer Nano ZS. The critical micelle concentration (CMC) was determined by pyrene fluorescence probe method. And the transcellular process of the micelles on MDCK epithelial cells was investigated by using transmission electron microscope, laser confocal scanning microscope and Förster resonance energy transfer technology. It turned out that the size of PEG-PCL micelles was about 30 nm and CMC was 1.01 microg x mL(-1). PEG-PCL micelles were endocytosed in intact form and they could deliver hydrophobic drugs across the basolateral membrane of the epithelial cells. However, PEG-PCL is hardly being transported in micelle formation itself. The transportation of C6 by PEG-PCL micelles was through the transcellular pathway, yet not the paracellular pathway.

iRGD-modified sterically stabilized liposomes loaded doxorubicin (iRGD-SSL-DOX) were prepared and their cellular toxicity and anti-tumor efficacy were evaluated, comparing to doxorubixin loaded sterically stabilized liposomes (SSL-DOX) and RGD modified doxorubixin loaded sterically stabilized liposomes (RGD-SSL-DOX). The iRGD peptide, with both tumor targeting and cell penetrating functions, was conjugated to DSPE-PEG-NHS and DSPE-PEG-iRGD was obtained. DSPE-PEG-RGD was gained in the same way. iRGD-SSL-DOX, RGD-SSL-DOX and SSL-DOX were prepared by ammonium sulfate gradient method. The size and zeta potential of the liposomes were characterized by dynamic laser light scattering. The cellular toxicity study was done on B16 melanoma cell line and the anti-tumor efficacy study was carried on B16 cell line bearing C57BL/6 mice. The results showed that the particle sizes of liposomes were all around 90-100 nm. DOX entrapment efficiency was above 95%. The formulations were with good preparation reproducibility. iRGD-SSL-DOX showed no significant difference in B16 cellular toxicity with SSL-DOX and RGD-SSL-DOX, but the anti-tumor efficacy on B16 melanoma bearing C57BL/6 mice was significantly better than that of SSL-DOX, similar as that of RGD-SSL-DOX. Therefore, iRGD modified liposomes loaded DOX would be a promising drug delivery system for tumor therapy.

In order to verify whether p-nitrophenylalanine-containing BAFF vaccine can be used as a candidate molecule for the treatment of autoimmune diseases with BAFF over-expression,a soluble mutant of B cell activating factor belonging to the TNF Family (smBAFF) and its pNO2Phe mutant(pNO2Phe65smBAFF),which site specific incorporated pNO2Phe at position 65 of smBAFF,were expressed and purified.In order to evaluate the feasibility of using pNO2Phe65 smBAFF to treat BAFF-over-expressed autoimmune diseases,we investigate its Lymphocyte-stimulating capacity,immunogenicity and inhibitory effect of serum on biological activity of natural BAFF.The pharmacological activity of pNO2Phe65 smBAFF was evaluated using a cGVHD(graft-versus-host disease) induced SLE mouse model.Results indicated that pNO2Phe65 smBAFF,could bind to mouse lymphocytes but could not promote the proliferation of mouse lymphocytes.Moreover,the incorporation of pNO2Phe significantly increased the immunogenicity and induced cross-antibody,which can inhibit the biological activity of natural BAFF.In cGVHD induced SLE mouse model,pNO2Phe65 smBAFF can significantly reduce the symptoms of the disease and play a therapeutic role.Therefore,pNO2Phe65 smBAFF can be used as a candidate molecule for the treatment of autoimmune diseases with BAFF over-expression.

Objective To explore impact of BMI on the clinical efficacy of treatment of type 2 diabetes mellitus after the laparoscopic gastric bypass. Methods Twenty patients were randomly selected from type 2 diabetes mellitus patients with BMI of 25 ~28 kg/m2 after laparoscopic gastric bypass surgery in our hospital from 2010 to 2013,20 patients from type 2 diabetes mellitus patients with BMI of 28~35 kg/m2 and 20 patients from type 2 diabetes mellitus patients with BMI≥35 kg/m2 . Accordingly,the total of 60 patients were divided into low BMI group,middle BMI group and high BMI group. The difference of the rate of blood glucose control and other indicators between three groups were compared and analyzed. Results The rate of blood glucose control in low BMI group was 75%,middle BMI group was 85%, and high BMI group was 90%. There was no statistical difference of the rate of blood glucose control between three groups at 12 months after laparoscopic gastric bypass surgery. The blood glucose and weight obviously improved in all patients,and fasting insulin and postprandial ser-um insulin decreased at 12 months after laparoscopic gastric bypass surgery. Conclusion All the patient of type 2 diabetes mellitus with dif-ferent BMI in three group can acquire satisfying clinical therapeutic effect after the laparoscopic gastric bypass.

Objective To evaluate the efficacy of itraconazole oral solution for prevention of invasive fungal infection ( IFI ) in neutropenic patients with acute leukemia after chemotherapy.Methods Clinical data of 136 neutropenic patients with acute leukemia after chemotherapy at the Department of Hematology,Nanfang Hospital from January 2008 to December 2010 were retrospectively analyzed.Patients were divided into itraconazole group ( n =67 ) and control group ( n =69).There were 36 patients with acute nonlymphocytic leukemia ( ANLL),31 with acute lymphoblastic leukemia (ALL) in itraconazole group;while in control group,there were 30 patients with ANLL,38 with ALL and 1 with biphenotypic acute leukaemia (BAL).Patients in itraconazole group received intraconazole after chemotherapy until the neutrophil count was increased to 0.5 × 109/L or the body temperature returned to normal and without any imaging evidence of IFI.The incidence of IFI and clinical features were compared between the groups using SPSS 13.0 software.Pearson x2 test was used for nominal variables,for measurement data,t (normal distribution) or Mann-Whitney U (skewed distribution) test were used.Results There were 12 cases ( 17.9％ ) suffering from IFI in itraconazole group and 32 cases (46.4％) in the control group (x2 =12.59,P ＜ 0.01 ).For ANLL patients,the incidence of IFI in itraconazole group was significantly lower than that in control group ( 16.7％ vs.56.7％,x2 =11.53,P ＜0.01 ).In itraconazole group,the incidence of IFI in female patients was significantly lower than that in male patients ( 8.6％ vs.28.1％,x2 =4.35,P ＜0.05 ).And for the female patients,the incidence of IFI in itraconazole group was significantly lower than thatin the control group (8.6％ vs.44.7％,x2 =11.98,P＜0.01).Conclusion Itranconzole oral solution can effectively prevent IFI in neutropenic patients with acute leukemia after chemotherapy,especially for the female patients with ANLL.

Objective To discuss effect of synchronous families cognitive intervention on mental, behavioral characteris-tics in cervical cancer. Methods Selected 60 cases with cervical cancer were divided randomly into three groups, each of 20 cases. Group 1 was nursed by routine care, group 2 was nursed by cognitive intervention only for patients, and group 3 was nursed by cognitive intervention for patients and families. Anxiety, depression and behavioral characteris-tics of patients before and after intervention were surveyed by SAS, SDS and C type behaviors scale. Results After in-tervention, SAS and SDS of group 3 were lower than group 2, and group 2 was lower than group 1, which showed sig-nificant difference (P<0.05 or P<0.01). After intervention, anxiety dimensions, depression dimension of group 2 were lower than group 1, and group 3 were lower than group 2 (P<0.01); optimism dimensions, social support dimensions of group 2 were higher than group 1, and group 3 were higher than group 2 (P<0.01 or P<0.05). Conclusion Synchronous families cognitive intervention for patients with cervical cancer can improve anxiety, depression and behaviors.

In order to delete the immune tolerance of self-protein HER2 and produce an efficient immune response,the genetic code expansion technique was employed to introduce the immunogenic amino acid p-nitro-phenylalanine into the position 5 ,26 and 79 of the HER2 fragment and HER2 mutants containing p-nitropheny-lalanine were obtained.Prototype and mutant HER2 molecules with a purity of more than 95% were obtained through a nickel affinity column.Immunogenicity analysis of the HER2 vaccine showed that pNO2Phe79HER2 was able to produce high titer cross-reactive antibodies in C57BL/6 mice and that the mouse antisera could recognize SKBR-3 cells with intact HER2 receptor extracellular domain and cleave HER2 highly expressed HER2 +B16F10 cells through ADCC effect.These findings suggested that pNO2Phe79HER2 which incorporates p-nitro-phenylalanine could be used as a tumor vaccine candidate targeting HER2.

OBJECTIVE: To investigate the inhibition effect of total alkaloid, total phenolic acid and total volatile oil from Ligusticum chuanxiong on the contraction of the isolated uterine smooth muscle of rats, and to provide reference for clarifying the promoting blood circulation to restore menstrual flow traditional efficacy of L.chuanxiong. METHODS: The isolated uterine smooth muscle of rats were collected and soaked in Locke's solution. Using dimethyl sulfoxide as blank control, verapamil hydrochloride as positive control, the contraction curves of the isolated uterine smooth muscle in rats were recorded with PowerLab electrophysiolograph. Inhibitory effects of total alkaloid, total phenolic acid (mass concentrations of 0. 025, 0. 05, 0. 1 mg/mL) and total volatile oil (0. 04, 0. 08, 0. 16 mg/mL) on the contraction of the isolated uterine smooth muscle of rats induced by oxytocin were evaluated by observing the changes of contraction activity and contraction tension. Inhibitory rates of contraction activity and contraction tension of uterine smooth muscle were calculated. RESULTS: The contraction activity and contraction tension of uterine smooth muscle in rats were increased significantly after treated with oxytocin (P<0. 05 or P<0. 01). Different concentrations of 3 types of compounds from L. chuanxiong all decreased contraction activity and contraction tension of uterine smooth muscle significantly (P<0. 05 or P<0. 01). Compared with blank control, except for 0. 025 mg/mL total phenolic acid, the inhibitory rates of contraction activity and contraction tension of uterine smooth muscle were all decreased significantly (P<0. 05 or P<0. 01)after treated with other concentration drugs or components. CONCLUSIONS: The total alkaloid, total phenolic acid and total volatile oil from L. chuanxiong show certain inhibitory effect on the contraction of isolated uterine smooth muscle in rats, which provide reference for promoting blood circulation to restore menstrual flow use of L. chuanxiong.

Objective BAG3-related myopathy is a rare condition so far reported in twenty patients worldwide.The purpose of this study was to draw attention to this rare disease and to the fact that BAG3-related myopathy should be considered as a rare differential diagnosis of hypercapnia.Methods We report a sporadic case of a 14-year-old Chinese girl with a de novo p.Pro209Leu mutation in BAG3 and reviewed the literatures for reported cases related to this mutation.Results We described a 14-year-old Chinese girl who presented with gradually appearing symptoms of hypercapnia that required assisted ventilation.The muscle biopsy and the blood whole-exome sequencing results confirmed the diagnosis of myofibrillar myopathy with a de novo p.Pro209Leu mutation in BAG3.Totally twenty-one patients from twenty families with a confirmed diagnosis of BAG3-related myopathy were reported to date,including this patient and literature review.The male to female ratio was 11 :10 and most showed initial symptoms in the first decade of life.Most patients presented toe/clumsy walking or running as the onset symptom,followed by muscle weakness or atrophy.Creatine kinase levels were elevated in fourteen patients and were normal in three.Eighteen patients developed respiratory insufficiency during the disease course and thirteen (one could not tolerate non-invasive assisted ventilation) required non-invasive assisted ventilation for treatment.Except for one not reported,heart involvement was found in seventeen patients during the disease course and seven underwent heart transplantation.Z-disk streaming and aggregation could be observed in most of the patients' muscle histology.In the long-term follow-up,five patients died of cardiac or respiratory failure.Conclusion BAG3-associated myopathy is a rare type of myofibrillar myopathy.It should be considered as a rare differential diagnosis of hypercapnia.