Objective To establish a method for the determination of eugenol in Xiao’er Fuxie Waifu Gel.Methods RP-HPLC method was developed with Lichrospher-C18 analysis column,using methyl alcohol-water(65:35) as mobile phase.The detection wavelength was 280 nm and flow rate was 1.0 mL/min.Results The linear range of eugenol was from 0.108 to 1.724 ?g(r= 0.999 9).The average recovery was 99.1 %and RSD was 1.4 %.Conclusion The method is convenient and accurate.It can be used for the quality control of Xiao’er Fuxie Waifu Gel.

Objective:To construct and evaluate a novel tissue-engineered bone composed of murine stromal cell-derived factor 1(mSDF-1), simvastatin (SIM) and collagen scaffold (Bio-Oss?), serving as a cell-homing approach for bone formation .Methods: In the study , 32 ICR mice were randomly divided into 4 groups,each group including 8 mice.The drug-loaded collagen scaffolds were implanted subcutaneously onto the cranium of each mouse according to the groups: ( 1 ) 1 ∶50 ( volume ratio ) dimethyl sulfoxide ( DMSO ) /phosphate-buffered saline ( PBS ) solution +collagen scaffold ( blank control group ); ( 2 ) 10 -3 mol/L SIM solution +collagen scaffold ( SIM group ); ( 3 ) 200 mg/L mSDF-1solution +collagen scaffold (mSDF-1 group); and (4) 10 -3mol/L SIM +200 mg/L mSDF-1 solution +collagen scaffold ( SIM +mSDF-1 group) .One week after implantation , the mice were trea-ted by injecting the same drug solution mentioned above around the scaffold once a day for two days .The specimens were harvested 6 weeks after implantation and the bone formation was evaluated by soft X-ray analysis , HE staining and immunohistochemical staining .Angiogenesis of each group was checked by calculation of vessels in each tissue section .Results:Six weeks after implantation , the collagen scaffolds were retrieved.The value of gray scale for the SIM +mSDF-1 group[(421 836.5 ±65 425.7) pixels] was significantly higher than that of the blank control group [(153 345.6 ±45 222.2)pixels, P<0.01], the SIM group [(158 119.2 ±100 284.2) pixels, P<0.01], and the mSDF-1 group[(255 529.5 ± 152 142.4) pixels, P <0.05 ]; HE staining analysis revealed that significant bone formation was achieved in the SIM +mSDF-1 group; The immunohistochemical staining showed the existence of os-teopontin and osteocalcin in the SIM +mSDF-1 group; There were more vessels in the SIM +mSDF-1 group[(46 ±8)vessels/mm2] than in the blank control group [(23 ±7) vessels/mm2, P<0.01], and the SIM group[(24 ±6) vessels/mm2 , P<0.01].Conclusion:The novel tissue-engineered bone com-posed of mSDF-1, SIM and collagen scaffolds has the potential to form bone subcutaneously in vivo.It re-presents a novel method of in vivo bone re-generation without seed cell delivery .

Objective To evaluate the value of non invasive examinations in the diagnosis of vascular disease of lower extremity. Methods Segmental systemic pressure (SEG), pulse volume recorder (PVR) and continuous wave (CW) were adopted to examination of 36 patients with arterosclerosis obliterans of lower limbs, and the foundings were compared with that of MRA and operation . Results In all 36 cases, non invasive examinations detected abdominal aorta and common iliac artery with femoral artery occlusion in 12 cases ,femoropoplitea artery occlusion in 28 cases, popliteal artery and the distal artery occlusion in 16 case; MRA together with operation confirmed abdominal aorta and common iliac artery with femoral artery occlusion in 11 cases, femoropopliteal artery occlusion in 32 cases and popliteal artery and the distal artery occlusion in 13 cases. The accurent diagnosis rate of non invasive examination was 85.7%. Conclusions Although PVR has not yet been used to replace the arteriography to directly observe the local situation of arterial lesions , it can preliminarily determine the obstruction sites and severe degree of the occluded arteries. PVR has convenient and reliability in the diagnosis,evaluation of the therapeutic effect and the postoperative follow up of blood vessel diseases,it is worthy to be widely applied in the clinical practice.

ObjectiveTo explore the characteristics and correlation of intelligence development and socioemotional development in 1~3 years old children with expressive language disorder,and provide a basis for early intervention.Methods125 cases with expressive language disorder(language disorder group) and 126 normal children (normal group) are evaluated in two parts (intelligence and social mood) with 0~6 years old neuropsychological development diagnosis scale and Chinese version of urban infant-toddler social and emotional assessment.Results (1)The scores in language disorder group were lower than normal group in the area of free movements (81.60±10.40 vs 89.62±7.94),adaptability (81.48±10.95 vs 91.25±8.89),language (67.46±7.20 vs 89.13±8.24),social behavior (76.61±9.73 vs 90.12±8.13) and total developmental quotient (80.17±6.39 vs 91.15±6.05) (P<0.05).(2) There were significant differences between language disorder group and normal group in the area of externalizing behavior(56.28±10.15 vs 53.57±7.91),deregulation (56.45±10.61 vs 51.11±9.32) and capabilities(46.79±9.08 vs 51.25±7.47) (P<0.05).The boys' scores in internalizing behavior were lower than the girls' (49.19±10.76 vs 54.71±9.90) (P<0.05).(3) The scores of gross motor had positive correlation with externalizing behavior (r=0.220,P<0.05).The scores of language had negative correlation with externalizing behavior and deregulation(r=-0.650,P<0.05;r=-0.470,P<0.05).The scores of social behavior had negative correlation with externalizing behavior (r=-0.208,P<0.05).There was also a negative correlation between total development and deregulation (r=-0.184,P<0.05).(4) Multiple stepwise linear regression results showed that the externalizing behavior domain and mothers' education could predict 44.1% of variances in the language area.ConclusionThe children with expressive language disorder not only show backward in level of language development,but also in the development of other areas,and also bad social behavior and emotional problems.Individualized parenting interventions should be adopted to promote these children's intelligent development in an all-round way.

Objective To investigate the effectiveness of ilioischial plating through modified Stoppa and iliac fossa approaches for complex acetabular fractures. Methods A consecutive series of 40 patients with complex acetabular fracture were treated operatively from January 2014 to February 2015. Of them, 20 were treated through modified Stoppa plus iliac fossa approaches as an experimental group ( including 12 males and 8 females with a mean age of 46. 8 ± 10. 3 years ) . The anterior column was stabilized with a recon-struction plate for the iliac wing along the iliopectineal line to the pubis in all cases. The posterior column was fixed with a newly developed ilioischial plate running from the ilium to the ischial ramus. The other 20 patients ( 10 males and 10 females with a mean age of 45. 6 ± 11. 7 years ) served as a control group, treated with a reconstruction plate for the conventional posterior column fixation through the Kocher-Langenbeck approach. The 2 groups were compared in terms of operative time, intraoperative blood loss, reduction and functional recovery of the hip. Results The 40 patients obtained an average follow-up of 18 months ( from 8 to 24 months ) . The experimental group reported significantly shorter operation time ( 2. 1 ± 0. 7 hours ) and signifi-cantly less intraoperative bleeding ( 320. 8 ± 100. 4 mL ) than the control group ( 2. 9 ± 0. 6 hours and 621. 6 ± 118. 7 mL, respectively ) ( P <0. 05 ) . According to modified Matta's criteria for reduction, the experimental group had 15 excellent, 3 good, one fair and one poor cases ( giving an excellent to good rate of 90％) while the control group had 17 excellent, one good, one fair and one poor cases ( giving an excellent to good rate of 90％) . According to the modified Merle d'Aubigné and Postel scoring for the functional recovery of the af-fected hip at the final follow-ups, the experimental group had 14 excellent, 3 good, 2 fair and one poor cases ( giving an excellent to good rate of 85％) while the control group had 12 excellent, 4 good, 3 fair and one poor cases (giving an excellent to good rate of 80％). There were no significant differences between the 2 groups in the above comparisons ( P> 0. 05 ) . There were no significant differences in the MOS item short form health survey score and postoperative complication rate between the experimental group and the control group. Conclu-sion Ilioischial plating through modified Stoppa and iliac fossa approaches has advantages of reliable fixa-tion, limited invasion, less intraoperative blood loss and fewer complications for complex acetabular fractures.

In order to determine the involvement of CALM-AF10 fusion transcripted in primary leukaemias with t(10;11) and its chemotherapy sensitivity in vitro, the AF10-CALM fusion transcripts were detected by reverse transcription-polymerase chain reaction (RT-PCR), and the chemotherapy sensitivity testing in vitro was undergone by MTT assay in five t(10;11) leukemia samples from patients with ALL, AML and lymphoblastic lymphoma. The results showed that five different-sized AF10-CALM product and four different-sized CALM-AF10 products were detected. The chemotherapy sensitivity of leukemic cells with t(10;11) in vitro to drugs is lower than that of leukemic cells without t(10;11). 3 out of 5 cases of t(10;11) leukemia were sensitive to chemotherapeutic drugs, while 31 out of 36 cases of leukemia without t(10;11) were sensitive at same condition. There were significant differences (P < 0.01), consistent with clinical features of patients. Apoptosis rate of leukemic cells with t(10;11) induced by chemotherapeutic drugs was lower than that of leukemic cells without t(10;11), (16.37 +/- 2.56)%, and (33.75 +/- 5.59)%, respectively (P < 0.01). It is concluded that the CALM-AF10 fusion transcripts are a common features and are involved in the pathogenesis of haematological malignancies with t(10;11), and are associated with a poor prognosis.
Antineoplastic Agents ; pharmacology ; Cell Survival ; drug effects ; Chromosomes, Human, Pair 10 ; genetics ; Chromosomes, Human, Pair 11 ; genetics ; Humans ; Leukemia ; genetics ; pathology ; Oncogene Proteins, Fusion ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transcriptional Activation ; drug effects ; Translocation, Genetic ; Tumor Cells, Cultured

Lactobacillus casei was selected as an antigen delivery vehicle for the development of oral vaccine to express recombinant LTB and porcine parvovirus (PPV) VP2 protein. The fusion protein gene encoding PPV VP2 protein and LTB, was cloned into the surface expression vector pPG, and then the recombinant expression vector pPG-VP2-LTB was electrotransformed into Lactobacillus casei 393, generating recombinant strain pPG-VP2-LTB/L. casei 393. After induced by 2% Lactose in MRS broth, an about 78 kD protein was detected in the recombinant Lactobacillus casei by SDS-PAGE. The result of Western blot indicated that the protein possessed the antigenic specificity same as the native virus protein. The result of the whole bacteria cell ELISA indicated that the LTB protein was expressed at the same time. The results of indirect immunofluorescence test and immuno-gold electron microscopy showed that the interest protein was expressed on the surface of L. casei 393. The results provide potential for the development of lactic acid bacteria oral vaccine of PPV, which used LTB as mucosal adjuvant.

OBJECTIVETo explore the immune response of silicotic rats to sheep red blood cells(SRBC).

METHODSSilicotic rats were immunized with SRBC by tracheal instillation(Group 1) or intraperitoneal injection (Group 2), and non-silicotic rats were immunized by tracheal instillation as normal control(Group 3). The levels of serum hemolytic index(HC50) were measured on 7, 12, 20, 25, and 32 days after primary immunization and 5, 12, 15 days after the second immunization. Special anti-SRBC IgG was measured with ELISA(A490 nm) on 12, 20, 25, 32 days and 5, 12, 15, 27 days respectively. Delayed-type hypersensitivity(DTH) to SRBC was measured 20 days after second immunization and DTH reaction was determined at 24, 48, 72, and 96 h after administration. Total cell count and cell populations in the bronchoalveolar lavage fluid(BALF), lung associated lymph node(LALN) and spleen weight, special IgG secreted from spleen cells were measured at the end of the experiment.

RESULTSThe HC50 of Group 1(47.4 +/- 1.0, 52.2 +/- 4.6, 31.1 +/- 11.9, 43.8 +/- 3.5, 33.6 +/- 16.8, 49.0 +/- 2.3, 92.9 +/- 20.2, 87.7 +/- 5.2) were statistically higher than those of Group 3(40.4 +/- 10.6, 2.8 +/- 2.5, 0.8 +/- 0.6, 6.6 +/- 5.8, 1.4 +/- 0.1, 36.5 +/- 16.5, 53.0 +/- 33.2, 2.6 +/- 2.2). The special anti-SRBC IgG response in Group 1(1.67 +/- 0.19, 1.98 +/- 0.36, 1.12 +/- 0.50, 1.38 +/- 0.30, 2.75 +/- 0.15, 2.60 +/- 0.28, 2.86 +/- 0.10, 2.50 +/- 0.20) were much stronger than those in Group 3 (0.59 +/- 0.30, 0.56 +/- 0.21, 0.21 +/- 0.16, 0.22 +/- 0.01, 0.81 +/- 0.25, 0.74 +/- 0.25, 0.69 +/- 0.26, 1.38 +/- 0.41). Furthermore, the results of DTH showed positive response and the ratios for diameter of skin rash > 5 mm at 24, 48, 72, 96 h were 16/16, 16/16, 16/16, 15/16 respectively in Group 1, while those in Group 3 were 8/15, 1/15, 1/15, 1/15 respectively. Total cell count in the BALF, LALN and spleen weight, and special IgG secreted from spleen cells in Group 1 were higher too. Group 2 expressed almost of the same but with mild immunologic responses as Group 1.

CONCLUSIONSilicosis-induced extremely strong DTH and over-response of humoral immunity to some antigens may contribute to the likelihood of silicosis complicated with tuberculosis.

Animals ; Erythrocytes ; immunology ; Hypersensitivity, Delayed ; etiology ; Immunization ; Immunoglobulin G ; blood ; Rats ; Sheep ; Silicosis ; immunology

OBJECTIVETo investigate restoration of alveolar cleft with engineered bone constructed by sponge collagen protein combined bone mesenchymal stem cells (BMSC).

METHODSTwelve dogs were divided into 4 groups, the third incisor and alveolar bone with periosteum in bilateral maxilla were removed to form alveolar cleft model. The BMSCs were isolated from dog bone marrow. After being cultured and induced, the BMSCs were seeded in sponge collagen protein and cultured for 48 hours. The composites of BMSCs and collagen were implanted into the defect of alveolar cleft. After 12 weeks' feeding, those dogs were sacrificed. Three-dimensional CT and histological examination were used to observe the progress of bone formation.

RESULTSThe defects healed at 12 weeks after being implant BMSCs-collagen composites, the width of engineered bone is resembled with positive control, but the height is less than positive control.

CONCLUSIONSThe engineered bone can restore the defect of alveolar bone effectively, it can be used clinically to treat alveolar cleft.

Alveolar Process ; abnormalities ; Alveoloplasty ; methods ; Animals ; Bone Marrow Cells ; cytology ; Bone Regeneration ; Cell Culture Techniques ; Cleft Palate ; surgery ; Collagen ; Dogs ; Female ; Male ; Mesenchymal Stromal Cells ; cytology ; Tissue Engineering ; methods

At present time, the widely used hepatitis B virus( HBV) vaccines consist of only the small hepatitis B surface antigen expressed in yeast or CHO cells. The frequency of non-responders to these vaccines has increased the demand for a more immunogenic vaccine. Some studies have suggested that the addition of preS region to the vaccine will improve its efficacy. However, the large protein (L) containing the whole preS region can not be effectively expressed in vitro. To overcome this problem, two chimeric contructs, SS1, surface gene containing preS1 region at C-terminus and SS2, surface gene containing preS2 region at C-terminus, were constructed and effectively expressed in our previous studies. Here we further constructed an expression vector containing both SS1 and SS2 expression cassettes by separation and ligation the SS2 cassette to a linearized SS1 expression vector pAO815-SS1. The recombinant vector was transformed into Pichia pastoris GS115 by electroporation. A high-level expression strain (GS115-SS1S2) was established by primary screening for His+ transformants and further analysis for induction products. ELISA results demonstrated that the expressed protein had S, preS1 and preS2 antigenicities simultaneously. Western blotting showed that the product can bind to all of the three antibodies, anti-S, anti-preS1 and anti-preS2. The expression protein was present in the form of particles of 20-35 nm diameter and the yield of recombinant particles reached 300-600 mg/L by fermentation. The SS1 and SS2 polypeptides kept intact in purified particles, suggesting that the stability of preS region has been significantly improved.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Epitopes ; genetics ; immunology ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis B Vaccines ; immunology ; Peptide Fragments ; genetics ; immunology ; Pichia ; genetics ; Protein Precursors ; genetics ; immunology ; Vaccines, Synthetic ; immunology