OBJECTIVE:To prepare polyclonal antibody for recombinant hirudin Ⅲ (rHV3). METHODS: The antigens were prepared through two typical methods: one was conjugated with rHV3 itself, another was coupled with bovine serum albumin (BSA). Conjugation results were detected through electrophoresis. Then the rabbits, guinea pigs and rats were immunized and with serum sample taken for determination of antibody titer by double immunodiffusion test. RESULTS: Antibodies were produced in rabbits and DHP guinea pigs after immunization using rHV3-BSA as antigen, with antibody titer of 1∶16. CONCLUSION: After being immunized with rHV3-BSA, rabbits and DHP guinea pigs can produce polyclonal antibody for rHV3.

Objective To explore the role of probiotics in the prevention of diarrhea in critically ill patients.Methods The patients receiving enteral nutrition from May 2009 to September 2010 in the Respiratory Intensive Care Unit were divided into routine feeding group (control group,n =38) and probiotic yogurt feeding group (yogury group,n =42).The control group accepted routine enteral nutrition liquid (Nutrison),while the yogurt group accepted conventional feeding plus probiotic yogurt.The incidence of dianhea,length of hospital stay,and other indicators were compared between these two groups.Results The incidence of diarrhea was lower in the yogurt group on the 5th,6th,and 7th days (P＜0.05).The cases of positive stool bacterial and fungal culture results in the yogurt group were less than those in the control group (P ＜ 0.05).In addition,the hospital stay was shorter [(13.17±3.79) d vs.(15.18±4.39) d,P=0.031],the cost was lower [(6307±2358) yuan vs.(8253±3186) yuan,P =0.002],and the satisfaction of the patients were higher in the yogurt group.Conclusion Probiotics may reduce the occurrence of diarrhea associated with enteral nutrition in critically ill patients.

Objective To study the ultrasonic features of fetus deformity.Methods 34 cases with fetus defomity were examined by Aloka 280 B-mode ultrasound and Hp-image point system(3.5 MHz transducer).Results Of the thirty-four cases with fetus deformity,thirty-one cases were seen and proved by the clinical induced labour and parturition.Only three of the cases were misdiangnosed.Conclusion Examination with the ultrasonic equipment is a simple and convenient way in the antenantal examination.Fetus deformity often exists simultaneously with various types of deformity and has close relationship with the amount of amniatic fluid.

Based on the accurate cognition about tumor pathology,relying on observation of clinic cases,combining unsettled problems about tumor,instructed by theory of pathogenesis,the project puts forward the cancerous toxin-rooted and rising and falling of vital qi and pathogen-determined tumor viewpoint.The purpose of this paper is to provide methods and theory for treatment of tumor.

ObjectiveTo observe the effect of the doctor-led telephone interview on the state of anxiety for outpatients with epilepsy.Methods100 outpatients with epilepsy and state of anxiety were divided into two groups: intervention group (doctor-led telephone interview) and control group (outpatient face-to-face visit in the epileptic clinic), 50 cases in each group. Doctor-let semi-structured telephone interview was performed in the intervention group, while routine face-to-face interview in outpatient was performed in the control group for 6 months. Self-rating Anxiety Scale (SAS) was used to assess the anxiety state before and 6 months after intervention. The state of anxiety and the patients' contents in the two groups were analyzed.ResultsThe scores of SAS in two group decreased 6 months after intervention and the intervention group was better than the control group.ConclusionThe doctor-led telephone interview can improve the state of anxiety for the patients with epilepsy.

Objective To investigate the changes of Th17 and Th22 cells in patients with Helicobacter pylori(HP)infection . Methods 66 cases of HP infection (HP infection group) and 30 cases of healthy people (control group) were selected .The levels of Th17 and Th22 cells in peripheral blood were measured by flow cytometry and the level of IL-17 and IL-22 in serum were meas-ured by ELISA .Results Comparing with the control group ,the levels of Th17 ,Th22 ,IL-17 and IL-22 were significantly increased in HP infection group(P<0 .05) .The correlation analysis showed that the levels of Th17 ,Th22 ,IL-17 and IL-22 were significantly positive correlated with DPM(P<0 .01) .Conclusion Th17 and Th22 cells were actived in patients with HP infection ,which caused the high expression of immune effector molecules ,and promoted the progress of immune damage .

Objective:To efficiently amplify NK cells and determine their cytotoxic activity against a variety of tumor cell lines in vitro,thereby providing evidence for potential clinical application.Methods: PBMCs were isolated from adult peripheral blood and co-cultured with K562 cells that were genetically modified to express 4-1BBL,IL-15 and IL-21 on the surface for 15 days to effectively amplify NK cells.The total cell number and Purity of CD3-CD56+ cells were measured.Granzyme B and perforin expression of the amplified NK cells were detected by flow cytometry and real-time PCR.The anti-tumor effect on different cancer cells was evaluated.Results: This method obtained a more than 1.1×1010 CD3-CD56+ NK cells with 95% purity over a 15 day amplification procedure.The expanded NK cells could efficiently release granzyme B and Perforin.The cytotoxicity against different tumor cells was followed the order from strong to weak:gastric,pancreatic,cervical,ovarian and renal cancer cells,with the highest activity against gastric cancer cell line A549 (90% at E∶T=10∶1) (P<0.05).A time-dependent killing effect of activated NK cells on cervical,liver and pancreatic cancer cells was observed.Conclusion: This amplification procedure can consistently generate large amounts of pure NK cells with effective cytotoxic function against a variety of tumor cells.

L-asparaginase activity produced by six E. coli J53 strains containing different plasmids and plasmid free E. coli J53 strain was compared. The enzyme activity of the plasmid- bearing strains was about 2—4 times lower than that of the plasmid free ones. Curing the R plasmids from E. coli J53, the activity of L-asparaginase increased and was close to that of the plasmid free strain. It is proved that the remarkable inhibition of L-asparaginase activity results from the presence of the plasmid in E. coli.

Objective To evaluate the value of dual energy CT with Lung Vessels application in diagnosing pulmonary embolism (PE) of children.Methods Forty-seven patients with nephrotic syndrome and aged ≤ 18 years old (32 male,15 female,mean age 15 years) underwent dual-energy CT pulmonary angiography (CTPA).Two radiologists recorded the presence or absence of emboli and counted the clots on a per-patient and per-lobe basis with Lung Vessels,respectively.With conventional CTPA as reference standard,sensitivity,specificity,positive predictive value (PPV),negative predictive value (NPV) of Lung Vessels application to detect PE were computed for two readers.Inter-reader agreement was also calculated.Results Conventional CTPA detected 51 clots in 35 lung lobes in 13 patients,giving PE incidence was 27.6％ (13/47) for this cohort.On a per-patient basis,readers 1 and 2 detected 25 and 21 patients with PE with Lung Vessels,corresponding to sensitivities,specificities,PPVs,and NPVs of 100.0％ (13/13),58.6％ (17/29),43.3％ (13/30),100.0％ (17/17) and 100.0％ (13/13),76.5％ (21/29),50.0％ (13/26),100.0％ (21/21),respectively.On a per-lung lobe basis,readers 1 and 2 detected 74 and 63 clots with Lung Vessels,respectively,corresponding to sensitivities,specificities,PPVs,and NPVs of 94.3％ (33/35),79.5％ (159/200),44.6％ (33/74),98.7％ (159/161) and 97.1％ (34/36),85.5％ (171/200),54.0％ (34/63),98.8％ (171/173),respectively.On a per-clot basis,134 and 105 clots were detected with Lung Vessels application,corresponding to sensitivities,specificities,PPVs,and NPVs for DECT of 96.1％ (49/51),64.9％ (159/245),36.3％ (49/135),98.7％ (159/161) and94.1％ (48/51),75.0％ (171/228),45.7％ (48/105),98.3％ (171/174)for readers 1 and 2,respectively.Excellent or good inter-reader agreement was observed on a per-patient,per-lobe and per-clot basis (Kappa =0.828,0.859,0.695 ; all P ＜ 0.01).Conclusions Dual-energyCT with Lung Vessels application has a high sensitivity and negative predictive value for PE detection in children.More clots are detected with Lung Vessels application than conventional CTPA,which deserve further investigation.

Objective To evaluate 3D CT features of normal anatomy,anatomic variations and fractures of occipital squama.Methods The 3D CT features on MIP,VR images were analyzed retrospectively in 589 pediatric cases.The normal anatomy,anatomic variations and fractures of occipital squama were observed respectively,and the differential diagnostic features including the individual location,appearance and extension were analyzed.Results Four hundred and thirty-three patients (75.2％) showed normal anatomy,including 154 patients with adult occipital anatomical features,279 patients with posterior intraoccipital synchondrosis,and 37 patients with Kerckring-supraoccipital synchondrosis.When cases with recent trauma history were excluded,113 patients (19.1％) showed anatomic variants,including unpenetrating sutures and penetrating sutures.The former could be subdivided to Mendosal sutures in 23 cases,superior median fissures in 19 cases,and midline supraoccipital fissures in 4 cases,while the latter could be subdivided to the interparietal bone variations in 54 cases,wormian bones in 23 cases,and accessory bones in 7 cases.Two or more variations coexisted in 33 cases.The occipital squama fractures were shown in 34 cases (5.6％),including linear fractures in 27 cases,comminuted fractures in 3 cases,with depression fracture in one case,separation of cranial sutures in 3 cases,and other fractures associated with variants in 3 cases.The fractures were sharp,or jagged,without limitation of the occification.Conclusion There are different 3D CT features of normal anatomy,anatomic variations and fractures of occipital squama in children,which are important for making the accurate diagnosis.