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

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.

BACKGROUND: Operative correction of hallux valgus is one of the most procedures in foot and ankle department, but postoperative plaster slab immobilization often brings great inconvenience and pain. Off-loading forefoot shoes can theoretically help them to do activities off bed early, and prevent loading on the forefoot during walking. But its advantages and disadvantages compared with plaster slab are still unknown.OBJECTIVE: To compare the clinical effects of over-the-toe plaster slab and off-loading forefoot shoes after hallux valgus correction.METHODS: Patients with hallux valgus undergoing distal Chevron osteotomy or Scarf osteotomy in the Huashan Hospital of Fudan University between May 2012 and October 2013 were retrospectively reviewed. Group A patients had a below-knee and over-the-toe plaster slab while Group B had an off-loading forefoot shoe applied after surgery.Radiological assessment was done at 6 weeks postoperatively. Immobilization was removed once bone union was indicated. Clinical scales and subjective satisfaction investigation were performed at final follow-up, and complications during follow-up were noted. All values between groups were analyzed statistically.RESULTS AND CONCLUSION: Totally 125 patients (157 feet) were completely followed up, including 83 patients (95 feet) in Group A and 42 patients (62 feet) in Group B. There was no significant difference in demographics, pre-and-post operative radiological assessments, clinical scale, union time or general satisfaction rates between two groups. However,Group B patients had less chance to suffer metatarsophalangeal joint or inter-phalangeal joint stiffness, restricted ankle motion and supra-ankle pains of operated side in comparison with Group A (P < 0.05), and were more willing to accept the same treatment if they could choose again (P < 0.05). Compared with the plaster slab, the application of off-loading shoes not only maintains correction level, but also reduces such complications as stiffness or restricted motion of the joint, limb pains and so on, which makes patients more likely to accept, and should be given priority to in the rehabilitation following hallux valgus correction.

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.

Objective To evaluate the effects of bone morphogenetic protein 2(BMP-2)gene modified tissue engineering bone combined with vascular bundle implantation in repairing segmental bone defect.Methods The isolated rabbit hone marrow stromal cells(MSCs),after being transfected by adenovirus carrying BMP-2 gene(Ad-BMP-2),were seeded on bovine cancellous bone scaffolds(BCB) to construct gone modified tissue engineering hone.The rabbit models with radial defects(2.0 cm long) were made and repaired with four methods including gene modified tissue engineering bone with vascular bundle implantation(Group A),gene modified tissue engineering bone(Group B),nongene modified tissue engineering bone with vascular bundle implantation(Group C),and only BCB scaffolds(Group D).After 4,8,and 12 weeks of operation,X-ray,histological examination,biomechanics analysis and capillary vessel ink infusion were conducted to observe angiopoiesis and osteogenesis.Results Group A gained better effect in the volume and activity of new bones than other groups,with vascular bundle sending out new branches into the transplanted bones and productive regeneration of capillary vessel.The defect in Group A was repaired satisfactorily.Group B showed better effect in speed and quality of bone formation than Group C under induction of BMP-2 gent.Mainly fibrous tissues but not new bones were observed in Group D.Conclusion BMP-2 gene therapy with vascular bundle implantation has very strong osteoinduction ability and quite good vascularization effect and is of great value to the treatment of bone nonunion and bone defects.

OBJECTIVE To observe the effect of electrolyzed oxidizing water(EOW) on the surface sterilization of indoor environment.METHODS Sterilized the surface of ground,wall,table,mob and thermostat-controlled water-bath in cell culture room and good laboratory practice(GLP) with EOW,then to draw the materials and culture with culture dish for 48 h in a 37℃ incubator,and count the colony number.Sterilization method and grouping: group A treated as a control,group B sterilized with EOW,group C sterilized with ultraviolate ray for 30 min and group D first treated with ultraviolate ray for 30 min,then sterilized with EOW.RESULTS In group A,the bacteria were overgrew and formed flakiness in 10/10 culture dishes;1 colony was formed in group B,the sterilization effective rate was 90%.The bacteria culture of group C found no bacteria growing after sterilization with ultraviolate ray,however,sample from surface and culture after sterilization were seen bacteria,though the number of bacteria was less than group A.The bacteria culture outcome of group D was negative.CONCLUSIONS The EOW has a good sterilization effect,it is safe and untoxic,costly cheap and convenient to use,and fit to claim of environmental protection.