Objective:To investigate the effect of IL-35 on inflammatory response and T cell response in allergic rhinitis.Methods: 37 patients(observation group) with allergic rhinitis and 35 healthy volunteers(control group) after allergen detection of allergic rhinitisin in our hospital from Jan 2012 to Jan 2016 were selected as study subjects.The peripheral blood of observation group and control group were collected,and the serum levels of IL-35 were detected by ELISA.The animal model of allergic rhinitis in mice was established,the peripheral blood of mice was collected,and the serum level of IL-35 and IgE were detected by ELISA.The eosinophils that infiltrated in nasal mucosa were detected after tissue biopsy in mice.The mouse spleen cells were isolated and the ovalbumin antigen was added in the culture medium,IL-35 was or was not added into the culture medium,the ovalbumin specific T cell responses was detected.The cytokines IL-2,IL-4,IL-5,IL-10,IL-13,IL-17,IL-23,IL-27 and TNF-α in culture supernatant of ovalbumin specific T cells were detected by ELISA.The expression of IL-2,IL-4,IL-5,IL-10,IL-13,IL-17,IL-23,IL-27 and TNF-α in ovalbumin specific T cells were detected by Real-time PCR.The activation of JNK,Erk1/2 and p38 signal pathway in ovalbumin specific T cells were detected by Western blot.Results: The serum level of IL-35 in observation group was significantly lower than control group(P<0.05).The results showed that the number of eosinophils which infiltrated in AR mice nasal mucosa was significantly higher than normal mice(P<0.05),while the serum level of IL-35 in AR mice was significantly lower than normal mice(P<0.05).Ovalbumin specific T cell reactivity assay showed that IL-35 could significantly inhibit the T cell response.ELISA and Real-time PCR results showed that IL-35 could significantly down regulate the expression of IL-4,IL-5,IL-13,IL-17,IL-23 and TNF-α,and up regulate the expression of IL-2,IL-10 and IL-27.The Western blot results showed that IL-35 can inhibit the activation of JNK,Erk1/2 and p38 signal pathway of ovalbumin specific T in cells.Conclusion: IL-35 can regulate the expression of inflammatory cytokines in inflammatory response and inhibit T cell response,thus reducing allergic rhinitis,the mechanism may be through regulation of JNK,Erk1/2 and p38 signal pathway activation.

Objective To explore the clinical manifestations of toxic epidermal necrolysis(TEN) and its rare pulmonary complications.Methods Clinical symptoms,treatment and prognosis of 1 child with TEN caused by carbamazepine were analyzed.Radiological images were reviewed to evaluate the manifestations and the outcome of chronic pulmonary complications associated with TEN.Results The patient had high fever shortly after a dosage increment of carbamazepine.A confluent erythematous exanthema developed rapidly into painful blistering with skin erosion,denudation and involvement of conjunctive and oropharyngeal mucosa.The diagnosis of TEN was made.The mucocutaneous damage was gradually recovered with steroid plus intravenous immunoglobulin for 3 weeks.However,the patient presented with respiratory failure in the recovery phase of TEN.The computer tomography revealed pulmonary bullae and pneumothorax in the right lung.Lung parenchyma was squeezed and pulmonary bullae ruptured with pneumothorax and atelectasis,which were absorbed gradually through thoracic drainages.The patient′s lung function and pulmonary bullae were partly improved during a 7-month follow-up.Conclusions TEN is a severe form of blistering skin di-sease which is characterized by an extensive loss of epidermis and mucous membrane.Chronic pulmonary complications may occur in recovery phase of TEN.Pulmonary bullae,which might be caused by mucous damage and respiratory obstruction,is a rare complication of TEN.

Objective To evaluate valvular functionality after transcatheter pulmonary valve replacement in sheep using a novel polymeric prosthetic pulmonary valve.Methods In this study,we designed a novel polymeric trileaflet transcatheter pulmonary valve with a balloon-expandable stent,and the valve leaflet was made of 0.1 mm expanded polytetrafluoroethylene (ePTFE).We chose bovine pericardium valve as control.Pulmonary valve stents were implanted in situ by right ventricular apical approach in 8 healthy sheep(6 for polymeric valve and 2 for bovine pericardium valve) weighing an average of(22.8 ± 2.2) kg.Angiography was performed after implantation to assess immediate valvular function.Color Doppler echocardiography and 64-row computed tomography were used to assess valvular function 4 weeks after implantation.Results Implantation was successful in 8 sheep.Angiography at implantation showed one polymeric valve was located below the ideal position and most of the stent was in the outflow tract of right ventricle.While,all the other prosthetic valves demonstrated orthotopic position and exhibited normal open and close functionality.Echocardiography 4 weeks after implantation showed all the prosthetic valves exhibited normal functionality and no significant insufficiency.The peak-peak transvalvular pressure gradient of the polymeric valves was (18.8 ± 6.0) mmHg,while that of two bovine pericardium valves were 9 mmHg and 20 mmHg.CT 4 weeks after implantation demonstrated orthotopic position of the stents except the above-mentioned one and all the stents had no deformation.Conclusion The success rate of transcatheter pulmonary valve replacement by right ventricular apical approach is satisfactory.The early valvular functionality of the novel ePTFE pulmonary valve after transcatheter pulmonary valve replacement in sheep is good.

The design space of the high shear wet granulation process was established and validated within the framework of quality by design (QbD). The system of microcrystalline cellulose-de-ioned water was used in this study. The median granule size and bulk density of granules were identified as critical quality attributes. Plackeet-Burmann experimental design was used to screen these factors as follows: dry mixing time, the impeller and chopper speed of dry mixing, water amount, water addition time, wet massing time, the impeller and chopper speed of wet massing and drying time. And the optimization was implemented with the central composite experimental design based on screened critical process parameters. The design space of the high shear wet granulation process was established based on the quadratic polynomial regression model. Since the P-values of both models were less than 0.05 and values of lack of fit were more than 0.1, the relationship between critical quality attributes and critical process parameters could be well described by the two models. The reliability of design space, illustrated by overlay plot, was improved with the addition of 95% confidence interval. For those granules whose process parameters were in the design space, the granule size could be controlled within 250 to 355 μm, and the bulk density could be controlled within a range of 0.4 to 0.6 g x cm(-3). The robustness and flexibility of the high shear wet granulation process have been enhanced via the establishment of the design space based on the QbD concept.
Cellulose ; chemistry ; Reproducibility of Results ; Technology, Pharmaceutical ; methods ; Water

OBJECTIVETo observe the inhibitory effect of gefitineb on the proliferation and its inducing effect on the apoptosis of mouse I-10 Leydig testicular cancer cells in vitro.

METHODSWe treated I-10 Leydig testicular cancer cells of mice with gefitineb at 0, 1.25, 2.5, 5, 10, 20, and 40 µmol/L. Then we determined the inhibitory effect of gefitineb on the growth of the cells by MTT, detected their early and late apoptosis by Annexin V-FITC/propidium iodide double staining and Hoechst 33258 nuclear staining, respectively, and observed the expressions of apoptosis-related proteins Bcl-2, Bax and caspase 3/9 by Western blot.

RESULTSCompared with the blank control group, gefitineb significantly inhibited the proliferation of the I-10 cells at 10 and 20 µmol/L (P < 0.05). The survival rate of the cells was (32.4 ± 2.8)% (P < 0.01) and their early and late apoptosis rates were (26.7 ± 4.2)% and (59.33 ± 10.2)% in the 40 µmol/L group, significantly different from those in the control (P < 0.05 and P <0.01). In comparison with the blank control group, gefitineb at 10, 20, and 40 µmol/L increased the expression of pro-apoptotic protein Bax by (41.9 ± 7.1), (60.1 ± 9.8), and (69.0 ± 11.3)% (all P < 0.05), decreased that of apoptosis-inhibitory protein Bcl-2 by (50.3 ± 8.9), (63.9 ± 6.9), and (88.7 ± 13.9)% (all P < 0.05), and elevated that of the cleft proteins caspase-3 by (69.0 ± 6.9)% (P < 0.05), (71.5 ± 8.1)% (P < 0.05), and (110.9 ± 14.2)% (P < 0.01) and caspase-9 by (51.8 ± 4.9), (54.7 ± 6.7), and (43.8 ± 11.8)% (all P < 0.05).

CONCLUSIONGefitineb can increase the cytotoxicity of I-10 Leydig testicular cancer cells of mice and induce their apoptosis via the mitochondria-mediated apoptosis signaling pathway.

Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Proliferation ; drug effects ; Cell Survival ; Leydig Cell Tumor ; drug therapy ; metabolism ; pathology ; Male ; Mice ; Neoplasm Proteins ; metabolism ; Neoplasms, Germ Cell and Embryonal ; drug therapy ; metabolism ; pathology ; Quinazolines ; pharmacology ; Testicular Neoplasms ; drug therapy ; metabolism ; pathology ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo observe the peri-implant osseointegration after the application of exogenous nerve growth factor (NGF)-gelatin sponge (GS) composite.

METHODSSix beagle dogs were used to establish bi-mandible simultaneous implant model after the first and second premolar extraction. Then the dental implants were implanted into the distal socket while the bone defect was made in the mesial socket of each mandible. Then the NGF-GS was implanted into the defects as NGF-GS group, the gelatin sponge alone was implanted as GS control and the control group was left empty. Two dogs were sacrificed each time at 4 weeks, 8 weeks and 12 weeks postoperatively. Specimens were subjected to general observation, radiography, bone histological and histomorphometric analysis for the new bone formation. The data were analyzed with SPSS 11.5 software package.

RESULTSThe bone density in the defects around implants at 4 and 8 weeks postoperatively was lower than the normal bone. The bone-implant contact ratio in the NGF-GS group [(57.7 ± 6.4)%] was significantly higher than that in the GS control group and the control group [the ratio were (44.2 ± 3.3)% and (31.2 ± 3.1)%] (P < 0.01) at 4 weeks postoperatively, the bone-implant contact ratio in the GS control group was also significantly higher than that in the control group (P < 0.01) at that time. The bone-implant contact ratio in the NGF-GS group [(94.8 ± 7.7)%] was slightly higher than that in the GS control group and the control group [the ratio were (83.0 ± 4.1)% and (86.4 ± 6.3)%] at 8 weeks postoperatively, but there was no statistical difference (P > 0.05). The bone density in the defects around implants at 12 weeks was almost the same as the normal bone, there was no difference of the bone-implant contact ratio.

CONCLUSIONSNGF-GS application could increase new bone formation, accelerate maturation of trabecular bone around the implants and shorten the period of osseointegration.

Animals ; Dental Implantation, Endosseous ; methods ; Dental Implants ; Dogs ; Gelatin Sponge, Absorbable ; therapeutic use ; Male ; Nerve Growth Factor ; therapeutic use

OBJECTIVETo investigate the role of N-cadherin and fibronectin during chondrogenesis.

METHODSImmunohistochemical method and autibody induced changes of aggregation of cells were used to assay the expressions of N-cadherin and fibronectin during cell differentiation.

RESULTSThe N-cadherin was present in the area of the cell nodular area in the 24 hours group after adding chondrogenic revulsant, then there was a down-regulating trend. Fibronectin was expressed in 48 and 72 hours groups after adding chondrogenic revulsant, and showed to be negative afterward. The antibody against fibronectin or N-cadherin could inhibit the formation of cellular nodule markedly.

CONCLUSIONSCell adhesion factors play an important role during cell differentiation. TGF-beta(1) stimulates chondrogenesis via transition from an initial N-cadherin-contributing stage to a succedent fibronectin-contributing stage during the process of chondrogenesis in MSCs. Further study is needed to evaluate whether or not it can promote chondrogenesis by transfecting cDNA of CAMs to MSCs.

Objective To study the relationships among resilience, self-awareness, personality, stress level and mental health in the warship soldiers. Methods Resilience Scale for Adults ( RSA), Eysenck Personality Questionnaire ( EPQ), Self Acceptance Questionnaire( SAQ), General Self-Efficacy Scale( GSES), Wallance Self Concept Scale (WSCS), Psychological Stress Self-evaluation Test (PSET) and Symptom Checklist (SCL-90)were used to survey 1451 warship soldiers. Results ① 2.5％ of the warship soldiers reported obvious psychological stress. Mental health of warship soldiers was worse than the norm of China population, except for obsessivecompulsive and interpersonal sensitivity factor. While the scores of other factors were significantly higher than the norm of Chinese soldiers(P＜0.01). ②The total scores of SCL-90, psychological stress and resilience, internal/external had negative correlation with self-awareness, and positive correlation with neuroticism. Compared to the SCL-90-negative group,SCL-90-positive soldiers (SCL-90 total score ＞ 160) had higher scores of nervousness and PSET and lower scores of RSA, internal/external and self-awareness (P ＜ 0.01 ). ③Regression analysis showed that stress levels, emotional stability,self-awareness and resilience were able to predict 35.1％ of mental health in warship soldiers. Mental health, emotional stability, internal/external and resilience were able to predict 33.2％of individual's stress level. Conclusion To some extent, warship soldiers have psychological stress and mental health problems. Resilience, personality, self-awareness are important factors affecting psychological stress and mental health.

Aim To investigate whether necroptosis mediates chemical hypoxia-induced HT22 mouse hippocampal cell injury and inflammation.Methods HT22 hippocampal cells were exposed to cobalt chloride (CoCl2) to establish a model of the chemical hypoxia-induced injury and inflammation.The expression level of RIP3 (an index of necroptosis) was determined by Western blot.Cell counter kit-8 (CCK-8) assay was used to test the cell viability.Lactate dehydrogenase (LDH) activity in the culture medium was measured with commercial kits.Mitochondrial membrane potential (MMP) was examined by rhodamine123 staining followed by photofluorography.The intracellular level of reactive oxygen species (ROS) was detected by 2', 7'-dichlorfluorescein-diacetate (DCFH-DA) staining followed by photofluorography.The secretion levels of interleukin-1β (IL-1β) and tumor necrosis factor-a (TNF-α) were measured by ELISA.Results Treatment of HT22 hippocampal cells with 600 μmol·L-1 CoCl2 for 36 h markedly induced cytotoxicity, leading to a decrease in cell viability to (52.0±2.65) % , indicating that chemical hypoxia-induced cellular injury model was successfully set up.Besides, CoCl2 induced considerable injuries and inflammation, evidenced by increases in LDH activity, ROS production, MMP loss, as well as the secretion levels of IL-1β and TNF-α.Co-treatment of the cells with 40~100 μmol·L-1 Nec-1 (a specific inhibitor of necroptosis) and CoCl2 markedly attenuated the decrease in viability induced by CoCl2, reaching the best anti-cytotoxicity inhibitory effect at 80 μmol·L-1.Meanwhile, the co-treatment with 80 μmol·L-1 Nec-1 blocked the above injuries and inflammatory response induced by CoCl2.In addition, treatment of HT22 hippocampal cells for 6~48 h up-regulated the expression of RIP3, and Nec-1 alleviated the up-regulation of RIP3 expression level induced by CoCl2.Conclusion Necroptosis mediates chemical hypoxia-induced HT22 hippocampal cell injury and inflammation.

Objective To investigate the relationship of TCRCα-575A/G polymorphism with anti-neutrophil antibody(ANCA) associated vasculitis in Chinese Han population. Methods 86 cases of ANCA associated vasculi-tis in Chinese Han population and 196 healthy subjects were enrolled. TCRCα-575A/G was genotyped by PCR-re-striction fragment length polymorphism (PCR-RFLP) assay. Case-control study was performed. Results No signifi-cant difference was found in either genotype distribution(AA,AG,GG) or allele frequencies between 86 patients and healthy subjects(P>0.05);But significant differences between AA group, AG group, and GG group in systolic pres-sure[(127.47±24.18)、(124.11±25.21)、(148.92±19.23) mm Hg],diastolic pressure [(75.35±14.12)、 (74.50±13.01)、(85.46±9.40) mm Hg],red blood cell count[(3.41±1.01)×109/L、(3.46±1.04)× 109/L、(2.68±0.67)×109/L] and hemoglobin [(90.45±20.69)、(100.66±29.80)、(77.61±15.81) g/L (P<0.05 for each) were found. The patients in GG group had higher blood pressure and more severe anaemia;By following the patients about (16.0±36.8) months,no statistics significance was found between groups with and without chronic renal failure in distributions and genetypes of TCRCα-575A/G (P>0.05 ). Conclusions In Chi-nese Han population,TCRCα-575A/G polymorphism might not be related to genetic susceptibility and chronic renal failure of ANCA associated vaseulitis;but G allele might be associated with more serious anaemia and hypertension.