Objective To identify high?risk groups of Charcot foot( CN) in the people with diabetic foot neuropathic ulcerations( NU) . Methods Twenty cases patients with CN who were diagnosed in General Hospital of the Chinese People Air Force from June 2008 to June 2013 and 58 patients with diabetic neuropathic ulcer who were hospitalized from January 2010 to December 2011 and followed up until June 2014 without foot deform?ity were retrospectively analyzed. All patient's general condition, examination and laboratory results, diabetic chronic complications,complication,diabetes distribution of foot ulcers,and plain features. Results There were no statistically significant differences in terms of patients' average age, sex ratio, proportion of smokers, BMI, HbA1c,blood lipid,dorsalis pedis artery diameter and diabetic nephropathy (Ⅲ?Ⅳperiod) ,chronic kidney dis?ease stage 3 above,proliferation diabetic retinal pathological changes,the prevalence of coronary heart disease between the two groups(P>0. 05). Compared with NU group,patients with single high proportion(40. 00%(8/20) vs. 10. 34%(6/58)),Short duration of diabetes((12. 37±5. 64) years vs. (14. 27±8. 04) years),Feet long numbness(6(5,9) years vs. 4(2,20) years),low rate of hardening of the arteries narrow(ABI<0. 9)( 0 ( 0/20) vs. 39. 66%( 13/58) ) ,high recurrent diabetic foot ulcer prevalence( 70. 00%( 14/20) vs. 25. 86%( 15/58)),more patients with diabetes mellitus autonomic neuropathy(75. 00%(15/20) vs. 39. 66%(23/58)),less combined with hypertension ( 25. 00%( 5/20 ) vs. 58. 62%( 34/58 ) ) , the differences were significant ( t orχ2=6. 981,2. 259,4. 068,3. 887,12. 405,7. 436,6. 724;P<0. 05) . Diabetic foot wound distribution on mesopodi?um of CN group and NU group was 36. 84%(7/19),6. 90%(4/58) respectively,the difference was significant (χ2=11. 443,P=0. 003) . Diabetic foot amputation rate( Wanger 4,5 grade) of CN group and NU group was 44. 44%(4/9),6. 90%(2/29) respectively,the difference was significant(χ2=4. 732,P=0. 020). Conclusion The characteristics of high?risk groups of diabetics Charcot foot in the people with diabetic foot neuropathic ulcerations are middle aged,no foot of ischemia,combine the diabetic autonomic neuropathy and the feet always with recurrent ulcers.

Objective To investigate the prevalence of depression among the patients with diabetic foot and analyze the influence factors. Methods One hundred and ten patients with diabetic foot were inquired and assessed with patient health questionnaire for self-administered measurement (PHQ-9), meanwhile, the demo-graphic data, metabolic data and diabetes behaviors were also investigated. Results Prevalence of depression was 47.3%. Logistic regression analysis showed that alternation of diarrhea and astriction (OR = 6.901, P =0.017) and formication (OR = 23.401, P = 0.009) were risk factors, and medical insurance (OR = 0.217, P =0.007) was a protective factor. Conclusions Depression is a frequent mental disorder in patients with diabet-ic foot and its influence factors include alternation of diarrhea and astriction , formication and medical insur-ance .

Objective To investigate the factors related to healing of severe diabetic foot gangrene (Wagner 4 class above) infected with pan-resistant Pseudomonas aeruginosa,and to guide clinical treatment.Methods Forty-nine hospitalized patients with diabetic foot gangrene (Wagner 4 class above) from January 2009 to July 2014 were enrolled.The affected foot wound secretion culture was pan-resistant Pseudomonas aeruginosa.According to the wound healing time,they were divided into wound healing group (26 cases,healing time ≤ 3 months) and wound un-healing group (23 cases,healing time ＞ 3 months).The general information,clinical indicators and treatment between two groups were compared,and the factors related to healing was analyzed by multi-factor unconditioned Logistic regression analysis.Results Compared with those in wound un-healing group,the blood flow volume of dorsal artery of affected foot and negative pressure attraction rate in wound healing group were higher:(43.59 ± 2.71) ml/min vs.(23.14 ± 5.39) ml/min,76.9％ (20/26) vs 47.8％(11/23),and the urinary micro-albumin was lower:(67.01 ± 3.32) mg/L vs.(234.03 ± 6.71) mg/L.There were significant differences (P ＜ 0.05 or ＜ 0.01).Multi-factor unconditioned Logistic regression analysis showed that the factors related to healing of severe diabetic foot gangrene infected with pan-resistant Pseudomonas aeruginosa were the blood flow volume of dorsal artery of affected foot (regression coefficient was-5.551,P =0.001),urinary micro-albumin (regression coefficient was 0.127,P =0.007) and negative pressure attraction (regression coefficient was-2.244,P =0.042).Conclusion The blood flow volume of dorsal artery of affected foot,urinary micro-albumin,negative pressure attraction are the factors related to healing of severe diabetic foot gangrene infected with pan-resistant Pseudomonas aeruginosa.

In order to obtain liposomes with properties of heptocyte-specificity and pH-sensitivity,four galactosylated derivatives were synthesized. A series of liposomes were prepared by mixing the galactosylated derivatives with DC-chol/DOPE respectively. The liposome 18-gal was proven to have favorable gene transfer efficiency to human hepatoma HepG2 cells, which was significantly inhibited in the presence of galactose solution, indicating that the liposomal transfection activity was mediated by asialoglycoprotein receptors. The liposome showed prominent pH-sensitivity and low cytotoxicity. Its optimum gene transfer conditions were also determined. The results showed that the liposome may be developed as a potential hepatocyte targeting pH sensitive delivery system for nucleic acid drugs.

Objective To study the effects of the monoclonal anti idiotypic antibody NP30 active immunization on egg granuloma formation and hepatic fibrosis in Schistosoma japonicum infection. Methods ICR mice were actively immunized with NP30 100 ?g ?3 ip. every 10 days while the mice in control group were injected with SP2/0 ascites ip. simultaneously. After cercariae challenging,the mice were killed at the 4th, 8th,12th, 16th, 20th and 24th week, respectively.Mouse livers were removed and stained histochemically with VG and subjected to immunohistochemical assay of collagen type Ⅰ,Ⅲ and fibronectin(FN).The volume of egg granulomas and the content of collagen type Ⅰ,Ⅲ and FN were determined quantitatively by NYD 1000 Image Analysis System. Results The volume of egg granulomas in NP30 immunized group was much smaller than that of control group from the 12th week after cercariae challenge. The cellular components of egg granulomas in NP30 immunized group were significantly different from those of the control group,exhibiting two types of atypical egg granulomas were found.VG stain revealed that the average optical density of collagen in hepatic granulomas of experimental group was lower than that of control group.Immunohistochemical assay revealed that the contents of collagen type Ⅰ,Ⅲ and fibronectin in egg granulomas of experimental group were lower than those of control group. Conclusion NP30 vaccination may induce both cellular and humoral protective immunity to modulate egg granulomas and suppress liver fibrosis of schistosomiasis japonica.

Objective To screen the mimic antigen epitopes of the triose phosphate isomerase of Schistosoma japonicum Chinese strain (SjC TPI) and investigate their immunogenicity. Methods The random phage peptide library (PH\^D\^ 12) was screened with the purified antibody(IgG) against SjC TPI to get the positive phage which contained the mimic antigen epitopes of SjC TPI, and the immuno characterization of the mimic antigen epitopes were investigated. Results Two mimic antigen epitopes (M1, M2) of SjC TPI were obtained. The immuno sera of mice (Kunming strain) against the positive phages could recognize both the SjC TPI and the protein of the positive phages. The DNA sequencing data showed no homology between the sequences of the deduced amino acid of the two mimic antigen peptides and the amino acid of SjC TPI. Conclusion The two mimic antigen epitopes of SjC TPI obtained are imitative epitopes of the configuration antigen of SjC TPI.

Objective To construct, express and purify human scFv antibody (S1) against the recombinant SAG1 of Toxoplasma fused to magainin, and observe its protective effect against Toxoplasma in infected mice. Methods The S1 scFv antibody gene amplified from phagmid S1/pIT-2 fused to magainin was cloned into procaryotic expression vector pET-32c. The recombinant plasmid S1M/pET-32c proved by DNA sequencing was transformed into E.coli BL21, and induced for fusion expression of S1M with IPTG. The expressed S1M was purified with Ni 2+ chelating HiTrap HP column and detected with SDS-PAGE. The effect of reduction of infection of Toxoplasma was observed through in vivo and in vitro experiments in mice. Results The fused gene of S1 and magainin was successfully cloned into procaryotic expression vector pET-32c proved by DNA sequencing. The recombinant S1M protein about 43 kDa was expressed in E.coli as inclusion body, and prepared with Ni 2+ column purification. Tachyzoite of Toxoplasma preincubated with S1M showed decreased infectivity in mice, the result of in vivo experiments showed that mice treated with S1M hadlonger survival time than the mice untreated. Conclusion The purified targeting antimicrobial peptide S1M could reduce the infectivity of tachyzoites of Toxoplasma in a certain extent and has a potential value for biological therapy of toxoplasmosis; otherwise, the constructed targeting antimic robial peptide S1M also provides a new model for biological therapy of toxoplasmosis.

Objective To develop a novel synergism compound suspension concentrate of niclosamide and chlorphoxim (Co-SCN) and sdudy its characteristics. Methods Niclosamide and chlorphoxim were milled by a ball mill and mixed with different amounts of wetting agent. disper-sant agent, thickener, and water etc. , to develop Co-SCN, and the pH value, thickener, grain size were evaluated. The ultraviolet absorption spectrum of niclosamide and chlorphoxim were measured. The content of niclosamide and chlorphoxim in the solution were assayed by HPLC. Results Co-SCN was a gray thickener fluidity liquid. It was very easy to disperse and could be mixed with water in any proportion. Its pH was 8. 65 and thickener was 137 mpa? s. The grain sizes (diameter) were from 0. 138-19. 953 ?m. Of them more than 95. 6% was smaller than 10 ?m and more than 82. 24% was smaller than 5 ?m. There were three peaks of ultraviolet absorption spectrum for niclosamide: 210, 234 nm and 334 nm respectively. One peak of chlorphoxim was at 269 nm. The novel formulation contained 20.64% niclosamide and 5.26% chlorphoxim. The suspension stability of Co-SCN was 100% for 2 hours and 89. 14% for 4 hours, and otherwise WPN in water was speedy sediment. Conclusion The novel synergism compound suspension concentrate of niclosamide and chlorphoxim is a stable quality and standard formulation.

Objective To enhance the protective immunity effects against Schistosoma japonicum infection by priming with cocktail DNA vaccines and boosting with cocktail protein vaccines in infected BALB/c mice.Methods Plasmids and proteins for immunization were prepared and diluted in no bacterial saline solution to final concentration of 1.5 mg/ml,and mixed with pcDNA3.1-SjC23,pcDNA3.1-SjCTPI,pcDNA3.1-(CDR3)6 plasmid DNAs by equal volume to form the cocktail DNA vaccine,and also mixed with recombinant proteins SjC23-HD,SjCTPI,and NP30 by equal volume to form the cocktail protein vaccine.Seventy female BALB/c mice of 4-5 weeks old were randomly divided into 5 groups(A,B,C,D,E).In Group A(control group),each mouse was immunized with 100 ?l saline solution by intramuscular(i.m.);in Group B(pcDNA3.1 control group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 for three times at week 0,3,6;in Group C(pcDNA3.1 and cocktail protein group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 for three times at week 0,3,6 and immunized with 100 ?l mixed protein vaccines plus 100 ?l FCA by subcutaneous at week 9;in Group D(cocktail DNA vaccines group),each mouse was immunized(i.m.)with 100 ?l mixed DNA vaccines for three times at week 0,3,6;in Group E(cocktail DNA vaccines plus cocktail proteins),each mouse was immunized(i.m.)with 100 ?l mixed DNA vaccines for three times at week 0,3,6 and immunized with 100 ?l mixed protein vaccines plus 100 ?l FCA by subcutaneous at week 9.Four weeks after the last DNA immunization or two weeks after protein boosting,all the mice were challenged with(40?1)cercariae of Schistosoma japonicum by abdominal skin penetration at the same time.Forty-two days post-challenge,the mice were sacrificed and perfused,and the numbers of recovered worms and eggs in liver were counted.The blood was collected from the tail veins of all the mice two days before the first immunization and challenge,respectively,the serum was prepared for detection of IgG,IgG1 and IgG2a.Two days before the challenge,the spleen cells of two mice from each group were cultured and stimulated with ConA and soluble egg antigen(SEA),and the supernatant was collected for detection of IL-2,IL-4 and IFN-?.Results The worm reduction rates in Group C,D and E were 17.70%,32.88% and 45.35%,respectively,compared with the control group.The worm reduction rates in Group D and E were significantly higher than that in Group C(P