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Objective:To explore the feasibility of applying plasma with same blood group as kidney donor to ABO incompatible kidney transplantation(ABOi-KT)preconditioning of blood group O recipients with high-titer anti-A/B preformed antibody(IgM/IgG titer ≥1∶256).Methods:A total of 15 cases of blood group O ABOi-KT recipients with high-titer anti-A/B were recruited and divided into two groups of AB( n=8)and kidney donor's blood(KD, n=7)according to plasma type for plasma exchange during preconditioning phase. Clinical data of preconditioning and post-KT were recorded. Results:They received plasmapheresis(PP)(8.1±2.5)sessions in preconditioning phase, including double plasma filtration(DFPP)(4.0±1.4)sessions and plasma exchange(PE)(4.1±2.0)sessions, PP frequency was(0.8±0.1)sessions per day. No hemolysis reaction occurred during preconditioning phase. Anti-A/B titers declined as expected and fulfilled the ABOi-KT criteria(IgM/IgG titers ≤1∶8). KT was performed successfully without antibody-mediated rejection. All of them survived with normal renal function within 90 days post-KT. Levels of serum creatinine at Day 7/30/90 post-KT were(92.9±30.4), (96.2±25.9)and(103.1±28.4)μmol/L; anti-A/B IgM titers at Day 7/30/90 post-KT 1∶1-1∶32, 1∶1-1∶64 and 1∶1-1∶32; anti-A/B IgG titers at Day 7/30/90 post-KT 1∶1-1∶64, 1∶1-1∶64 and 1∶1-1∶32 respectively. No significant differences existed in count/frequency of PP sessions, levels of serum creatinine or anti-A/B titers at each observation point between AB and KD groups( P>0.05). Conclusions:Plasma with the same blood group as kidney donor is feasible for maximizing the intensity of ABOi-KT preconditioning. Favorable outcomes may be achieved through an intensified desensitization strategy on blood group O recipients with high-titer anti-A/B preformed antibody. The potential risks and long-term outcomes should be further explored.
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Objective:To evaluate the effect of transcutaneous electrical acupoint stimulation (TEAS) on perioperative anxiety and postoperative pain in living kidney donors (LKDs).Methods:Seventy-two American Society of Anesthesiologists physical status Ⅰ or Ⅱ LKDs, aged 18-64 yr, with body mass index of 18-28 kg/m 2, undergoing living kidney transplantation, were selected, and divided into 2 groups ( n=36 each) using a random number table method: TEAS group (group T) and sham stimulation group (group S). In group T, TEAS was performed on the forenoon at 1 day before surgery (T 0), at 30 min before anesthesia induction on the morning of the operation day (T 1) and on the forenoon at 1 day after surgery (T 2) at bilateral Neiguan, Taichong and Yintang with a frequency 2-100 Hz, disperse-dense waves and current intensity 6-15 mA, and each TEAS lasted for 30 min.Only electrode patches were applied at the same acupoint and at the same time point, but no stimulation was applied in group S. In T and S groups, brachial venous blood samples were collected before each stimulation for measurement of the plasma 5-hydroxytryptamine (5-HT) concentration.The Hospital Anxiety Depression Scale-Anxiety subscale (HADS-A) scores at T 0, T 1, T 2, on day 3 after surgery (T 3) and before discharge (T 4) in the 2 groups were recorded.The consumption of anesthetics during operation, laryngeal mask airway removal time, requirement for rescue analgesia within 72 h after surgery and the development of postoperative complications were recorded.The LKDs were followed up by telephone at 3 months after surgery (T 5) to record the scores of HADS-A and Leeds Assessment of Neuropathic Symptoms and Sign (LANSS) scale. Results:Compared to group S, the incidence of anxiety was significantly decreased T 1, T 2 and T 3, the incidence of rescue analgesia within 72 h after surgery was decreased, plasma concentration of 5-HT was increased at T 1 and T 2, the incidence of postoperative nausea and vomiting was decreased, and the time to first flatus was shortened in group T ( P<0.05). There was no significant difference in the consumption of anesthetics during operation, laryngeal mask airway removal time, and the incidence of anxiety and neuropathic pain within 3 months after surgery between the 2 groups ( P>0.05). Conclusion:TEAS can relieve early preoperative and postoperative anxiety and alleviate postoperative pain in LKDs.
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Objective To explore the risk factors for lower extremity amputation in patients with diabetic foot.Methods The clinical data of 1 771 patients with diabetic foot at the Air Force General Hospital of PLA from November 2001 to April 2015 were retrospectively analyzed.The patients were divided into the non-amputation and amputation groups.Within the amputation group , subjects were further divided into the minor and major amputation subgroups.Binary logistic regression analyses were used to assess the association between risk factors and lower extremity amputation.Results Among 1 771 patients with diabetic foot , 323 of them ( 18.24%) were in the amputation group ( major amputation: 41; minor amputation:282 ) and 1 448 ( 81.76%) in the non-amputation group.Compared with non-amputation patients, those in the amputation group had a longer hospital stay and higher estimated glomerular filtration rate(eGFR)levels.Fasting plasma glucose (FPG), glycosylated hemoglobin (HbA1c), C-reaction protein (CRP), ESR, ferritin, fibrinogen and WBC levels of the amputation group were higher , while hemoglobin albumin, transferrin, TC, TG, HDL-C and LDL-C were lower than those of the non-amputation group (all P<0.05 ).The proportion of hypertension ( 52.48% vs 59.98%) , peripheral vascular disease ( PAD ) (68.11% vs 25.04%), and coronary heart disease (21.33% vs 28.71%) were different between the amputation and non-amputation groups (all P<0.05).Multivariable logistic regression analyses showed that Wagner′s grade , PAD and CRP were the independent risk factors associated with lower extremity amputation in hospitalized patients with diabetic foot.Conclusion Wagner′s grade, ischemia of lower limbs and infection are closely associated with amputation of diabetic foot patients.
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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 .
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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.
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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.
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Objective To establish a diagnostic scoring system for coronary artery disease(CAD)in patients with diabetic foot(DF)and evaluate its application efficiency. Methods The clinical data of 445 DF inpatients were analyzed retrospectively in this case?control study. These patients were divided into the CAD group(n=372,DF with CAD)and the control group(n=73,DF without CAD)according to the presence or absence of CAD. Risk factors were screened from related clinical factors examined through multiple logistic regression analysis for CAD in patients with DF and were assigned according to odds ratio(OR)to establish the scoring system for diagnosis of CAD in patients with DF. Application efficiency of the di?agnostic scoring system was tested by calculating area under the receiver operating characteristic(ROC)curve. Results The multiple logistic re?gression analysis showed that risk factors for CAD in patients with DF were age,male sex,the duration of diabetes≥10 years,the ankle?brachial in?dex(ABI)≤0.9,body mass index(BMI)≥25 kg/m2 and chronic renal insufficiency. According to ORassigned age(0.9=0,≤0.9=2),BMI(<25 kg/m2=0,≥25 kg/m2=3)and chronic renal insufficiency(absent=0,present=3)scores. Area under the ROC curve of the diagnostic score scheme was 0.758(0.682?0.835),the standard error was 0.039,and the point of the diagnosis of CAD was 7. Conclusion The scoring system established in the study is efficacious,simple and practical,which provides an important reference for CAD in patients with DF.
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Objective To make a cross-sectional assessment of the morbidity of lower extremity arterial disease (LEAD) in inpatients with type 2 diabetes mellitus (T2DM) and to analyze its risk factors,thus providing evidence for its clinical prevention.Methods We enrolled 664 inpatients with T2DM from June 2012 to June 2013 and collected clinical data,including age,gender,duration of diabetes,body mass index,smoking,fasting & postprandial blood glucose levels,glycosylated hemoglobin,serum lipids,renal function,fibrinogen,neck ultrasonography,lower extremity vascular ultrasound,ankle brachial index and treatment records.Logistic multiple regression analysis was conducted to identify risk factors for LEAD.Results A total of 247 cases met the diagnostic criteria for LEAD,with morbidity reaching to 37.2％.The percentages of morbidity in patients with different durations of diabetes were:23.12％ (≤ 5 years),27.95％ [(5 10) years],38.71％ [(1015) years],51.16％ [(15-20) years],62.34％ (≥ 20 years).The differences were statistically significant (P＜0.05).Of the patients in the LEAD group,73.2 ％ were treated with antihypertensive medications and 54.6％ were treated with statins.The goal attainment rates for total cholesterol,lowdensity lipoprotein cholesterol,high-density lipoprotein cholesterol and triglycerides were 56.3％,39.3％,47.4％ and 61.5％,respectively,in the LEAD group and 45.1％,34.5％,35％,and 49.4％,respectively,in the non-LEAD group.With the exception of the rates for low density lipoprotein cholesterol,the rates between the two groups are statistically significant (P＜0.05).Significant differences in age,BMI,blood pressure,coronary heart disease,cerebrovascular disease,carotid intima-media thickness,carotid artery plaque,and carotid artery stenosis were also observed between the two groups (P＜0.05 for all parameters).Logistic multiple regression analysis revealed that age,history of diabetes,cerebrovascular disease,carotid artery plaque,and carotid artery stenosis were risk factors for LEAD.Conclusions The morbidity of LEAD is 37.2％ in type 2 diabetic patients.Age,history of diabetes,cerebrovascular disease,carotid artery plaque,and carotid artery stenosis are risk factors for LEAD,while traditional risk factors for atherosclerosis,including hypertension,levels of cholesterol and low-density lipoprotein cholesterol,smoking,and non-drug intervention,are risk factors for LEAD in type 2 diabetic patients.
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Objective To investigate the clinical effects and indications of the vacuum sealing drainage (VSD) in the treatment of severe diabetic foot gangrene.Methods We randomly recruited 60 cases,who had suffered from diabetic foot gangrene(DFG) at the grade of 3 -5,according to Wagner scale into VSD treatment groups and treated them with VSD methods.At the same time,62 DFG cases who had given routine drainage treatment one year ago were retrospectively analyzed as control group.The observed items included the wound healing time,number of dressing,outcome of treatment ( healing rate),the average days in hospital,total expenses of hospitalization and so on.Results The wound healing time of VSD treatment group and routine treatment group were ( 30.5 ± 6.8 ) days and ( 53.8 ± 5.5 ) days,respectively ( t =2.636,P ＜ 0.01 ).The numbers of dressing were( 15.0 ± 4.7) days and ( 29.5 ± 6.1 ) days,respectively ( t =2.374,P ＜ 0.01 ).The healing rates were 96.7％ (58/60) and 87.1％ (54/62),respectively(P ＜0.01 ).The average period in hospitalization were (20.1 ± 3.5 ) days and ( 36.5 ± 4.6 ) days,respectively ( t =2.564,P ＜ 0.01 ).Total expenses of hospitalization were(20 155.6 ± 153.8) yuan RMB and(41 465.5 ± 146.6) yuan RMB,respectively(t =2.873,P ＜0.01 ).All the differences were statistically significant.Conclusion The VSD method is effective for the treatment of severe diabetic foot gangrene(DFG).It is able to reduce the time of wound healing significantly,increase the healing rate,shorten the hospitalization period and cut the general expenses during hospitalization.It' s an effective method for the treatment of DFG.
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Objective To construct a recombinant immunotoxin expression vector composed of a single-chain Fv fragment of Sehistosorna japomicum and PE38KDEL gene,and identify the binding activity of the purified product with SEA antigen.Methods The V_H and V_L genes were amplified by PCR from the parent monoclonal antibody NP11-4.Then the amplified scFv and PE38KDEL genes were inserted into the expression vector pBAD/gIII A.The fusion protein expressed in E.coli Top10F' induced by L-arabinose.After purification,the activity of the immunotoxin was evaluated by Westem-blot and ELISA.Results The new recombinant immunotoxin expression vector pBAD/gIII A-scfv-PE38KDEL was constructed successfully.The main product was in inclusion bodies.ELISA assay showed that the refolding recombinant immunotoxin remained binding activity with SEA antigen.Conclusion A new recombinant expression plasmid pBAD/gIII A-scfv-PE38KDEL has been constructed and expressed successfully,which is useful in further study of the treatment of schistosomiasis japonica.
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Objective To develop a special ambulance for airfield rescue of pilots in danger or offering medical care for pilots in flight.Of course the ambulance can also be used to rescue the wounded daily or in the war.Methods The ambulance owned a cross-country motorcar chassis and bearing carriage.A luffing extension-jib was installed on top of the carriage with a telescopic nacelle.The medical carriage owned a generator,air-conditioner,launder & antisepsis facilities,telescopic medical table,Air Force medical facilities for first aid and so on.It could simultaneously treat 2 injured pilots in lying position.Oxygen outputs were equipped to sustain Persons in carriage.Result The maximal speed of the ambulance was 95 km/h.The time of simulated rescue was about 3 minutes in the maxium height.Conclusion Without new staff established,the ambulance can adapt to any road and is suitable to war field.It can arrive at the location of flight accident quickly and rescue the pilot rapidly.It meets the needs of medical support in flight,war-time medical care in airport and mobile accompanying support.
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<p><b>OBJECTIVE</b>To investigate the protective immunity induced by the anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum in mice.</p><p><b>METHODS</b>An orthogonal table L(16) (4 x 2(12)) was selected as the experimental design. Eight-week-old Kunming outbred mice (male and female) were randomly divided into 16 experimental groups and 2 control groups. Control groups were injected with SP2/0 ascites intraperitoneally. Mice from each group were infected with 100 +/- 2 cercariae of Schistosoma japonicum in the abdominal skin and were sacrificed on the thirtieth day postchallenge. Adult worms were recovered and counted by perfusion of the left ventricle-portal vein. The SP2/0 ascites injected mice were used as controls and the percentage of protection was calculated.</p><p><b>RESULTS</b>Active immunization of mice with NP30 could produce protection levels ranging from 22.36% to 50.46% depending on the different immunity protocols. The best immunization protocol was established from the results.</p><p><b>CONCLUSIONS</b>Active immunization with NP30 can induce a degree of protection to infection with Schistosoma japonicum cercariae and NP30 is a potential vaccine candidate against Schistosoma japonicum.</p>
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Animals , Female , Male , Mice , Analysis of Variance , Animals, Outbred Strains , Antibodies, Anti-Idiotypic , Allergy and Immunology , Therapeutic Uses , Antibodies, Monoclonal , Allergy and Immunology , Therapeutic Uses , Schistosoma mansoni , Allergy and Immunology , Schistosomiasis mansoni , Allergy and Immunology , Parasitology , Treatment Outcome , VaccinationABSTRACT
Objective] To amplify and sequence the light chain of anti idiotypic monoclonal antibody NP30 of Schistosoma japonicum. [Methods] By comparing the conserved regions at each end of the nucleotide sequences of murine germ line genes enco ding FR1 and FR4 regions of immunoglobulin light chain variable regions, we designed a set of primers for amplification of V L gene. The hybridoma cells secreting anti idiotypic monoclonal antibody NP30 of Schistosoma japonicum were cultured and their genome DNAs were extracted and used as templates for PCR. The PCR product was then cloned into pUC19 vector. The recombinants were sequenced by Sanger′s method. The V L gene was compared with GenBank and published mouse V L genes. [Results] The full length of V L gene was 318 bp. The V L gene was a member of mouse Ig ? light chain subgroup IV and generated from rearrangement of germ line V and J? 4 genes. The V L gene sequence has been registered by GenBank(accession No. AF206720). [Conclusion] The obtained V L gene was a potentially functional gene of anti idiotypic monoclonal antibody NP30 of Schistosoma japonicum .
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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.
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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.
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Objective To enhance the protective immunity effects of nucleic acid vaccines against Schistosoma japonicum infection by electroporation(EP)in vivo 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.pcDNA3.1-SjC23,pcDNA3.1-SjCTPI,pcDNA3.1-(CDR3)6 plasmid DNAs were mixed 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/EP control group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 followed by EP in vivo for three times at week 0,3,6;in Group C(pcDNA3.1/EP plus cocktail protein vaccine group),each mouse was immunized(i.m.)with 100 ?l pcDNA3.1 followed by EP for three times at week 0,3,6 and boosted with 100 ?l cocktail protein vaccine plus 100 ?l FCA by subcutaneous at week 9;in Group D(cocktail DNA vaccine/EP group),each mouse was immunized(i.m.)with 100 ?l cocktail DNA vaccine followed by EP for three times at week 0,3,6;in Group E(cocktail DNA vaccine/EP plus cocktail protein vaccine group),each mouse was immunized(i.m.)with 100 ?l cocktail DNA vaccine followed by EP for three times at week 0,3,6 and boosted with 100 ?l cocktail protein vaccine 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.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-? by flow cytometre.Results The worm reduction rates in Group C,D and E were 18.09%,45.00% and 57.09%,respectively,compared with the control group.The worm reduction rates in Group D and E were significantly higher than that in Group C(P
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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
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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.
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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.
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Objective To construct single chain antibody specific to membrane protein of Schistosoma japonicum by gonetic engineering technique. Methods The V\-H (heavy-chain variable region) and V\-L (light-chain variable region) genes were amplified by PCR from the genomic DNA of NP11-4 cell line, and sequenced by Sanger's method. The ScFv was constructed in pTHA90 vector using V\-H and V\-L genes, then expressed by IPTG. Results The V\-H and V\-L genes were obtained through PCR. The DNA sequences showed that V\-H and V\-L were new variable region genes of antibody. They were registered by GenBank. A ScFv gene with (Gly4Ser) 3 intralinker in the pTHA90 vector was successfully constructed. The ScFv was expressed as thioredoxin-fused proteins about 36^2 kDa. Conclusion A specific ScFv against the membrane protein of Schistosoma japonicum was constructed and expressed.