Objective To construct the RNAi eukaryotic vector of inhibitory member of the ASPP family(iASPP) gene and observe the interfering effect in Jurkat cell line after the vector transfection.Methods The specific siRNA sequence was designed according to the iASPP sequence in GenBank.The sequence was cloned into PGCsilencer~(TM) H1/Neo/GFP and then sequence analysis was performed.The recombinant plasmid was transfected into Jurkat cell by liposome.The iASPP expression was analyzed by RT-PCR and cell apoptosis was detected by FCM.Results The sequence of templet and specific siRNA was correct by sequence analysis.The iASPP expression in Jurkat cells decreased and the apoptosis rate increased from 11.81% to 33.15% after RNAi transfection.Conclusion The RNAi eukaryotic vector PGCsilencer~(TM)H1/Neo/GFP/RNAi was constructed successfully.The RNAi inhibitory effect on the Jurkat cells is confirmed.The successful construction of iASPP RNAi makes it possible to further study the interaction between iASPP and p53.

To summarize the accumulated successful experiences, defeated cases and lessons the recent year’s clinical studies, works and experiences of hip prosthesis combining the relevant status abroad and to analyze some questions such as the operations are not standardized; the indications are not strict; overemphasizing the navigation and mini-invasion; clinical techniques are irregular, and the patients should be educated, and so on. The above questions influence the good development of the Chinese joint prosthesis and reduce the clinical outcomes of the joint prosthesis. Taken together, these questions should be concerned and resolved. Relevant solutions are discussed.

Objective To investigate the effect of once yearly zoledronic acid of 5 mg on postmenopausal women with osteoporosis of different causes. MethodsFrom October 2009 to December 2009,a total of 89 postmenopausal women with osteoporosis were enrolled and assigned into 2 groups.There were 45 cases of primary postmenopausal osteoporosis,including 27 cases of fresh fracture,in group A.They were aged from 47 to 83 years,with an average of 63.7 years.There were 44 cases of secondary postmenopausal osteoporosis,including 28 cases of fresh fracture,in group B.All patients were given a.single 30-minute intravenous injection of zoledronic acid (5 mg),supplemented by 1,25-dihydroxyvitamin D of 0.25 μg and calcium of 600 mg with VitD125 IU daily.At pre-intervention and 12 months after intervention respectively,bone mineral density (BMD) was measured by dual-X-ray absorptiometry (DXA) at the lumbar spine and hip,and a balance test(Sunlight Tetrax- Ⅱ) was performed to evaluate the risk of falling.Intervention compliance of the patients and adverse events related to zoledronic acid infusion were observed. Results All cases of fresh fracture healed well at 3-month follow-up.At 12 months,43 subjects in group A and 42 subjects in group B completed the follow-up.In group A,BMD increased by 5.8％ at the lumbar spine,by 2.9％ at the femoral neck,by 5.2％ at the Words area,by 5.3％ at the greater trochanter and by 3.9％ at the total hip while the risk of falling decreased by 26.1％; in group B,BMD increased by by 3.4％ at the lumbar spine,by 2.1％ at the femoral neck,by 3.2％ at the Words area,by 3.0％ at the greater trochanter and by 2.5％ at the total hip while the risk of falling decreased by 21.8％.The differences between pre-intervention and post-intervention were significant in both groups ( P ＜ 0.05).No intolerable adverse events occurred in both groups except that one new fracture happened in each group but responded to conservative treatment.ConclusionA once-yearly infusion of zoledronic acid of 5 mg is a convenient and effective therapy for treatment of osteoporosis in postmenopausal women.

BACKGROUND:Bone morphogenetic proteins are associated with the formation and development of scars. Fibroblasts are closely related to the proliferation and maturation of scars. However, the expression of bone morphogenetic proteins in fibroblasts in different stage of hypertrophic scars remains poorly understood.
OBJECTIVE:To observe the expression of bone morphogenetic protein 2/4, bone morphogenetic protein 7, bone morphogenetic protein receptor IA in fibroblasts in different stage of hypertrophic scars.
METHODS:The immunohistochemical SP method was employed to detect the expression of bone morphogenetic protein 2/4, bone morphogenetic protein 7, bone morphogenetic protein receptor IA in fibroblasts in 20 cases of proliferative stage hypertrophic scar and 20 cases of maturation stage hypertrophic scar. Al samples were obtained from the Department of Plastic and Aesthetic Surgery, the People’s Hospital of Guangxi Zhuang Autonomous Region, China.
RESULTS AND CONCLUSION:The expression of bone morphogenetic protein 2/4 in fibroblasts in proliferative hypertrophic scars significantly elevated compared with mature hypertrophic scars (P<0.05). In various stages of hypertrophic scars, the expression of bone morphogenetic protein 7 and bone morphogenetic protein receptor IA in fibroblasts was not obviously different (P>0.05). Results demonstrated that bone morphogenetic protein 2/4 expression down-regulated during the development from proliferative stage to maturation stages in fibroblasts in hypertrophic scars. However, there was no change in the expression of bone morphogenetic protein 7 and bone morphogenetic protein receptor IA.

Objective Construct the eukaryotic expression vector of inhibitory member of the ASPP family (iASPP) and trans-fect it into colon carcinoma cell lines SW480 and Lovo by liposome .Then observe the expression of iASPP and detect the cell apop-tosis by flow cytometry .Methods The amplified PCR product was digested and inserted into pMD19-T simple vector and sub-cloned into eukaryotic expression vector pcDNA3 .1(+ ) .The recombinant eukaryotic expression plasmid pcDNA3 .1(+ )-iASPP was transfected into colon carcinoma cell lines SW480 and Lovo by liposome ,the iASPP expression was analyzed by RT-PCR .The cell apoptosis was detected by FCM .Results The eukaryotic expression plasmid pcDNA3 .1(+ )-iASPP was constructed success-fully ,the gene squence of iASPP was consistent with that reported (gi 60457962) in GenBank .The mRNA expression levels of iASPP gene of SW480 and Lovo cell lines which transfect the positive plasmid were increased ,and the cell apoptosis rates were de-creased .Conclusion We successfully constructed the recombinant expression plasmid pcDNA 3 .1(+ )-iASPP ,and the plasmid were successfully expressed in colon carcinoma cell lines SW 480 and Lovo ,the cell apoptosis rates of those cell lines were decreased .These facts indicated that reducing the high expression of iASPP may be a new strategy to renew the abilities of P 53 tumor suppressor .

Objective To analyze the controlled risk factors of osteoporotic vertebral fractures, and determine the clinical value for the management of risk factors. Methods 626 cases of postmenopausal women were selected, age, height, weight and bone mineral density(BMD) of patients were collected.The Tetrax balance test system was used to assess the fall risks.Vertebral changes of patients were evaluated through X ray of thoracolumbar lateral. The clinical data were analyzed and compared between vertebral fracture group and non-vertebral fracture group. Spearman test was used to analyze the correlation between vertebral fracture and age, height, weight, body mass index(BMI), the risk of fall, body fat and BMD. Results 328 patients presented with osteoporotic vertebral fractures, while the other 298 cases did not suffer from osteoporotic vertebral fractures. There were 426 vertebral bodies involved. Comparing to non-vertebral fracture group, the vertebral fracture group showed higher age [(68.67±9.29)years vs.(63.04± 9.30)years], lower height[(151.10 ± 4.39)cm vs.(154.90 ± 5.86)cm], lower bone mineral density[lumbar spine BMD (0.85 ± 0.16)g/cm2 vs.(0.93 ± 0.17)g/cm2, hip spine BMD (0.72 ± 0.18)g/cm2 vs.(0.81 ± 0.13)g/cm2],higher body fat [(41.30 ± 5.20)％vs.(36.30 ± 5.90)％] and higher fall risk (41.38 ± 25.79 vs. 36.20 ± 26.22) ( P<0.001). While there were no statistical differences in weight and BMI between the two groups, age, height, BMI, body fat, fall risk, lumbar spine and hip BMD were significantly correlated with vertebral fracture (r=0.358,-0.323, 0.169, 0.186, 0.135,-0.398,-0.364, respectively, all P<0.001). Conclusions Decreased bone mineral density, increased age, abdomen fat content and increased fall risk, are the risk factors of osteoporotic vertebral fractures.

OBJECTIVEOn the basis of the concept of adhesive deformity from upper eyelid fold formation, the clinical results after using various methods to correct the adhesive deformities are summarized.

METHODSA total of 33 cases of adhesive deformity from upper eyelid fold formation have been treated using various corrective methods including taking off the sutures, shifting of the septal fat or the pre-septal orbicularis muscle, transferring of pretarsal orbicularis muscle, grafting of autogenous fat tissue, and repairing or/and shortening of the palpebral levator. Postoperative follow-up ranged from 3 months to 2 years.

RESULTSThe effective results have been got with the used methods except shifting of the pre-septal orbicularis muscle or transferring of pretarsal orbicularis muscle.

CONCLUSIONSThe suitable methods to correct the adhesive deformity from upper eyelid fold formation must be chosen according to the causes and the local situations.

Adult ; Eyelids ; abnormalities ; surgery ; Female ; Follow-Up Studies ; Humans ; Middle Aged ; Treatment Outcome

OBJECTIVETo investigate the effects of culture supernatant of human amnion mesenchymal stem cells (hAMSCs-CS) on biological characteristics of human fibroblasts.

METHODS(1) hAMSCs were isolated from deprecated human fresh amnion tissue of placenta and then sub-cultured. The morphology of hAMSCs on culture day 3 and hAMSCs of the third passage were observed with inverted phase contrast microscope. (2) Two batches of hAMSCs of the third passage were obtained, then the expression of vimentin of cells was observed with immunofluorescence method, and the expression of cell surface marker CD90, CD73, CD105, and CD45 was detected by flow cytometer. (3) hAMSCs-CS of the third passage at culture hour 72 were collected, and the content of insulin-like growth factor Ⅰ (IGF-Ⅰ), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), and basic fibroblast growth factor (bFGF) were detected by enzyme-linked immunosorbent assay. (4) Human fibroblasts were isolated from deprecated human fresh prepuce tissue of circumcision and then sub-cultured. Human fibroblasts of the third passage were used in the following experiments. Cells were divided into blank control group and 10%, 30%, 50%, and 70% hAMSCs-CS groups according to the random number table (the same grouping method below), with 48 wells in each group. Cells in blank control group were cultured with DMEM/F12 medium containing 2% fetal bovine serum (FBS), while cells in the latter 4 groups were cultured with DMEM/F12 medium containing corresponding volume fraction of hAMSCs-CS and 2% FBS. The proliferation activity of cells was detected by cell counting kit 8 and microplate reader at culture hour 12, 24, 48, and 72, respectively, and corresponding volume fraction of hAMSCs-CS which causing the best proliferation activity of human fibroblasts was used in the following experiments. (5) Human fibroblasts were divided into blank control group and 50% hAMSCs-CS group and treated as in (4), with 4 wells in each group, at post scratch hour (PSH) 0 (immediately after scratch), 12, 24, 48, and 72, the migration distance of cells was observed and measured with inverted phase contrast microscope. (6) Human fibroblasts were grouped and treated as in (5), with 3 battles in each group, and apoptosis rate of cells was detected by flow cytometer. Data were processed with analysis of variance of factorial design, analysis of variance for repeated measurement, one-way analysis of variance, LSD test, and t test.

RESULTS(1) On culture day 3, most hAMSCs were in large form, and spindle-shaped with much prominences like fibroblasts or in flat polygonal shape. hAMSCs of the third passage were spindle-shaped. The expression of vimentin of hAMSCs of the third passage was strongly positive, and the expressions of surface markers CD90, CD73, and CD105 of the cells were positive, while the expression of CD45 of the cells was negative. (2) The content of IGF-Ⅰ, VEGF, EGF, and bFGF in hAMSCs-CS were respectively (11.7±1.0), (316±68), (6.1±0.4), and (1.49±0.05) pg/mL. (3) At culture hour 12-72, the proliferation activity of human fibroblasts in each hAMSCs-CS group was significantly higher than that in blank control group (with P values below 0.01), and the proliferation activity of human fibroblasts in 50% hAMSCs-CS group was the highest. (4) The width of scratch in two groups was nearly the same at PSH 0. The migration distance of cells in 50% hAMSCs-CS group was significantly longer than that in blank control group at PSH 12-72 (with P values below 0.01). (5) The apoptosis rate of human fibroblasts in blank control group was (16.2±2.4)%, which was significantly higher than that in 50% hAMSCs-CS group [(7.4±3.6)%, t=6.710, P<0.01].

CONCLUSIONShAMSCs-CS can promote proliferation and migration of human fibroblasts and inhibit the apoptosis of human fibroblasts.

Amnion ; cytology ; Apoptosis ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Culture Media, Conditioned ; chemistry ; Enzyme-Linked Immunosorbent Assay ; Epidermal Growth Factor ; metabolism ; Female ; Fibroblast Growth Factor 2 ; metabolism ; Fibroblasts ; cytology ; drug effects ; Flow Cytometry ; Humans ; Insulin-Like Growth Factor I ; metabolism ; Male ; Mesenchymal Stromal Cells ; chemistry ; Pregnancy ; Vascular Endothelial Growth Factor A ; metabolism

Objective: To summarize the treatment experience of patients with different degree of acute respiratory distress syndrome (ARDS) caused by inhalation of white smoke from burning smoke bomb. Methods: A batch of 13 patients with different degree of ARDS caused by inhalation of white smoke from burning smoke bomb, including 2 patients complicated by pulmonary fibrosis at the late stage, were admitted to our unit in February 2016. Patients were divided into mild (9 cases), moderate (2 cases), and serious (2 cases) degree according to the ARDS Berlin diagnostic criteria. Patients with mild and moderate ARDS were conventionally treated with glucocorticoid. Patients with severe ARDS were sequentially treated with glucocorticoid and pirfenidone, and ventilator-assisted breathing, etc. were applied. The vital signs, arterial oxygenation index, changes of lung imaging, pulmonary ventilation function, general condition, and the other important organs/systems function were timely monitored according to the condition of patients. The above indexes were also monitored during the follow-up time of 10-15 months post injury. Data were processed with SPSS 18.0 statistical software. Results: (1) The symptoms of respiratory system of patients with mild and moderate ARDS almost disappeared after 3 days′ treatment. Their arterial oxygenation index was decreased from post injury day 1 to 4, which almost recovered on post injury day 7 and completely recovered one month post injury. The symptoms of respiratory system of patients with severe ARDS almost disappeared at tranquillization condition 1-3 month (s) post injury. Their arterial oxygenation index was decreased from post injury day 3 to 21, which gradually recovered 1-3 month (s) post injury and was normal 15 months post injury. (2) Within 24 hours post injury, there was no obvious abnormality or only a little texture enlargement of lung in image of chest CT or X-rays of patients with mild and moderate ARDS. One patient with moderate ARDS had diffuse patchy and ground-glass like increased density shadow (pulmonary exudation for short) at post injury hour 96. Chest iconography of all patients with mild and moderate ARDS showed no abnormalities 10 months post injury. Both lungs of each of the two patients with severe ARDS showed obvious pulmonary exudation at post injury hours 45 and 75, respectively. One patient with severe ARDS showed no abnormality in chest image 10 months post injury, but there was still a small mesh-like increased density shadow in double lobes with slight adhesion of pleura in the other patient with severe ARDS 15 months post injury. (3) All patients showed severe restrictive hypoventilation when admitted to hospital. Pulmonary ventilation function of patients with mild and moderate ARDS recovered to normal one month post injury, and they could do exercises like running, etc. Pulmonary ventilation function of one patient with severe ARDS recovered to normal 6 months post injury, and the patient could do exercises like running, etc. The other patient with severe ARDS showed mild restrictive hypoventilation 15 months post injury and could do exercises like rapid walking, etc. (4) The condition of all mild and one moderate ARDS patients was better on post injury day 3, and they were transferred to the local hospital for subsequent treatment and left hospital on post injury day 21. One patient with moderate ARDS healed and left hospital on post injury day 29. Patients with severe ARDS healed and left hospital on post injury day 81. During the follow-up time of 10-15 months post injury, the other important organs/systems of all patients showed no abnormality, and there was no adverse reaction of glucocorticoid like osteoporosis, femoral head necrosis, or metabolic disorder. Two patients with severe ARDS did not have any adverse reaction of pirfenidone like liver function damage, photosensitivity, anorexia, or lethargy. Conclusions Early enough and uninterrupted application of glucocorticoid can significantly reduce the ARDS of patients caused by inhalation of white smoke from burning smoke bomb. Sequential application of glucocorticoid and pirfenidone can effectively treat pulmonary fibrosis at the late stage.

BACKGROUND:Schwann cells are important celllines in the process of repairing peripheral nerve injury, and human amniotic homogenate supernatant is shown to secrete a variety of cytokines, which could promote the proliferation of Schwann cells.
OBJECTIVE:To investigate the effect of different concentrations of human amniotic homogenate supernatant on the proliferation of rat Schwann cell96.
METHODS:Schwann cell96 was cultured with high-glucose DMEM containing 20%fetal bovine serum, and the second generation of Schwann cell96 was applied for experiments. The cultured cells were divided into five groups according to different volume fractions of human amniotic homogenate supernatant (0%, 10%, 15%, 20%, 25%) in the medium.
RESULTS AND CONCLUSION:The total protein concentration of human amniotic homogenate supernatant was 675μg/mL, in which the concentration of epidermal growth factor, basic fibroblast growth factor and vascular endothelial growth factor were respectively (470.625±2.546), (4.121±0.026) and (0.172±0.002) ng/L. At 1-7 days, the cellproliferation rate of the 10%and 15%concentration groups was greater than that in 20%and 25%concentration groups (P<0.05);10%and 15%concentrations promoted cellproliferation, while 20%and 25%concentrations inhibited cellproliferation. There were no significant difference in the viability of Schwann cell96 between the control group and the experimental group (P>0.05). Low concentrations (10%, 15%) of human amniotic homogenate supernatant promote the proliferation of Schwann cell96, while high concentrations (20%, 25%) of human amniotic homogenate supernatant inhibit cellproliferation.