Objective To determine the involvement of DNA demethylation in tumor necrosis factor-α(TNF-α)-induced matrix metalloproteinase 9 ( MMP9) expression in human epidermal keratinocytes. Methods Real-time RT-PCR, Western blot, and enzyme-linked immuno sorbent assay (ELISA) were performed to determine the mRNA and protein levels of MMP9 after human keratinocyte cell line (HaCaT) cells were treated with 10 ng/ ml TNF-α or 2. 5 μmol/ L DAC/ 300 nmol/ L TSA. Bisulfite sequencing PCR ( BSP) and Methylation-sensitive high-resolution melt analysis ( Ms-HRM) were used to detect significantly differentially demethylated CpG sites in the human MMP9 promoter region in cells exposed to TNF-α. Different sites methylation constructs of promoter-luciferase reporter gene were made to detect the influences of site-specific DNA demethylation on transcription activity of MMP9 promoter. Results Compared with PBS-treated control, TNF-α significantly increased the expression of MMP9 in HaCaT cells for indicated culture duration ( P < 0. 05 ). Real time PCR, Western blot, and ELISA analysis demonstrated that the mRNA and protein levels of MMP9 were increased initially, followed by a decline with prolonged incubation time. After TNF-α treatment, varied degrees of DNA demethylation occurred at 10 CpG sites in the promoter of MMP9, and the changes at the -36 bp site were statistically significant (P<0. 05). The demethylation at the -36 bp site greatly increased the transcription activity of MMP9. Conclusion TNF-α promotes MMP9 expression in HaCaT cells through inducing -36 bp site DNA demethylation on the promoter of MMP9.

Adult ; Female ; Hemangioma, Cavernous ; surgery ; Humans ; Nose ; surgery ; Orbit ; surgery ; Orbital Neoplasms ; surgery

Objective To screen the valid plasmid targeted toGCN5 among the constructed recombinant plasmids;and to explore the effects of histone acetylizad modification in regulating MSCs differentiation.Methods Exstract the constructed plasmids and transfect them into MSCs induced by 5-aza.For 24 h,observe MSCs;detect the transfect efficiency by flow cytometry;detect expression of protein GCN5 by Western-blot;detect the HAT activity by ELISA.Results Transfect efficiency was more than 20%;Expression of protein GCN5 and HAT activity had no difference in group ZJ1 and ZJ2,but had a significant difference to that in group ZJ3.HAT activity of experiment group was significantly lower than that of control groups.Conclusion The inhibition state of histone acetylation results in plasmid ZJ3,it can inhibit the differentiation process of MSCs.The results provide data for the clinical application of MSCs transplantation.

Objective To investigate the prevalence of serum Anti-sperm antibodies(AsAb) in group of prepubertal boys with cryptor-chidism before and after surgery,and explore whether it may be influenced by testicular location,orchiopexy,unilateral or bilateral cryptorchi-dism and deferens and(or) epididymis abnormality.Methods Serum AsAb was examined in 50 prepubertal boys with cryptorchidism and 50 boys with oblique inguinal hernia before and 6 months after surgery.Fifty boys who were normal were served as controls.IgG and IgM AsAb were determined by the immunodotting method.Results The preoperative and post-orchiopexy serum AsAb positive rates in cryptorchid boys were significantly higher than the preoperative and post-high ligation of hernial sac ones respectively in boys with oblique inguinal hernia(Pa0.05).Conclusion Cryptorchidism may induce an autoimmune response against sperm antigen in prepuberty independent of testicular location,orchiopexy,unilateral or bilateral cryptorchidism and deferens and(or) epididymis abnormality.

OBJECTIVETo investigate the clinical effects of enblock distal radius T-plate fixation in treating trailing edge fracture of tibial plateau medial condyle and posterior cruciate ligament (PCL) avulsion fracture.

METHODSFrom September 2008 to November 2011, 5 patients with trailing edge fracture of tibial plateau medial condyle and PCL avulsion fracture caused by knee joint hyperextension,were treated with surgery. There were 3 males and 2 females with an average age of 35 years old (25 to 42). Left knee injury was in 3 cases and right knee had in 2 cases; 2 cases caused by sports injury and 3 cases caused by road accident. All patients were undergone emergency treatment. Fractures were anatomically reduced and fixed with T-plate through poples approach. Complications were observed after operation. Bone healing and clinical effects were respectively evaluated by X-rays and HSS system.

RESULTSFive patients were followed up for 1 to 4 years with a mean of 2 years. All incisions obtained healing of I stage. All knee joints recovered well and were stable with inflexion more than 120 degrees, no complications such as knee joint pain, injuries of nerve and blood vessel, infection, internal fixation failure were found. The mean score of HSS system was 94.40 +/- 6.09 and all patients got excellent result.

CONCLUSIONFor the treatment of trailing edge fracture of tibial plateau medial condyle and PCL avulsion fracture caused by knee joint hyperextension, enblock distal radius T-plate fixation had advantages of good stability,higher success rate, and knee joint function can recover well.

Adult ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; Humans ; Male ; Posterior Cruciate Ligament ; diagnostic imaging ; injuries ; surgery ; Tibial Fractures ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; Treatment Outcome

Air ; analysis ; Chromatography ; methods ; Iodine ; analysis ; Workplace

[Objective]To explore the relationship between peripheral arterial disease(PAD)and diabetic retinopathy(DR)in type 2 diabetes patients.[Methods]A total of 99 patients diagnosed with PAD were classified into grade 1-3 by their total scores of peripheral arterial stenosis assessed by color doppler ultrasound examinations,where the degree of stenosis 30% ~ 49% scored 0, 50%~99%scored 1,lumen occlusion(i.e. degree of stenosis 100%)scored 2,and therefore the total score 0-2 was categorized into Grade 1 ,3~4 into Grade 2 ,5~12 into Grade 3. The bilateral anterior tibial artery ,posterior tibial artery and dorsalis pedis artery of these patients were analyzed. The presence of diabetic retinopathy(DR)was graded from retinal photographs using a standard protocol.[Results]Among 99 cases of type 2 diabetic patients with peripheral arterial disease ,58.6%of them were male with average age of 67.3 ± 7.9 years old. Patients of Grade 1,Grade2,Grade 3 lesion accounted for 45.4%,30.3%,24.2%,respectively. Age, gender,smoking history,SBP,DBP,BMI,FBG,TC,TG,LDL-C,HDL-C,HbA1C among 3 groups were not statistically signifi-cant. The associations of DM duration and HbA1C value were significantly larger in DR than in PAD. The proportion of DR patients increased with the severity degree of PAD(p for trend=0.004). Degree of stenosis Grade 2 and Grade 3 could be predictive for DR.[Conclusions]DR is associated with the severity degree of PAD in type 2 diabetes patients as evaluated by duplex ultrasonography.Degree of stenosis Grade 2 and 3 could be used for screening or finding DR. Strategies for optimum treatment and early prevention are needed.

ObjectiveTo investigate the detection results of WBC,CRP,ESR and GR% in patients with knee infection after operation. Methods The study was conducted from March 2012 to January 2015. Twenty patients were collected in the study. The values of WBC,CRP,ESR and GR% were compared and analyzed between pre-operation and post-operation of 1,3 and 7 days. Results The values of WBC,CRP,ESR and GR% in the post-operation of 1 day were higher than those in the pre-operation,and there were statistically significances (P < 0.05). The values of ESR in the post-operation of 3 and 7 days were higher than that in the pre-operation, with statistically significances(P < 0.05). However,the values of WBC,CRP and GR% in the post-operation of 3 and 7 days were no statistically significances compared with those in the pre-operation(P > 0.05). Conclusion WBC,CRP,ESR and GR% have good reference value for early diagnosis and treatment,which can be used as screen testing indexes in the early knee infection.

Objective To observe the effects of genistein on PCNA expression and cell cycle in fibroblasts derived from human hypertrophic scar in order to explore the mechanism of its inhibition on hypertrophic scar (HS) fibroblast proliferation. Methods The human hypertrophic scar fibroblasts were cultured in vitro. Genistein with various concentrations (25, 50, 100 μmol/L) was co-cultured in the medium for 48 hours. The expression of PCNA was detected with immunocytochemical staining method and the cell cycle was measured with flow cytometry. Results Genistein could significantly decrease PCNA expression in HS fibroblasts, especially when its concentration at 50 μmol/L or 100 μmol/L. The cell percentage of G0～G1 phase decreased with drug′s concentration, and G2～M percentage increased conversely, implying the suspension of mitosis. In 100 μmol/L group, most cells blocked at S phase and a hypodiploid apoptosis peak could be observed ahead of G1 phase. Conclusion Genistein can inhibit the proliferation of human hypertrophic scar by blocking cell division as well as decreasing DNA synthesis.

Objective To analyze the present situation of iodine deficiency disorders (IDD) in Henan Province,and to promote implementation of sustainable control strategies.Methods In 2011,a stratified proportion to population probability sampling (PPS) method was used to survey 1200 children aged 8 to 10 in 30 counties of the province.One primary school was selected in each chosen county.Goiter,intelligence quotient (IQ),urinary iodine and salt iodine level were studied.Meanwhile,12 families per capita salt intake was investigated.In each school,30 5th-grade students and 30 pregnant and lactating women in the school townships and adjacent neighboring townships were selected to carry out questionnaire survey on health education with unified papers.Results ①The goiter rate of children aged 8 to 10 by B ultrasound was 4.5％ (54/1201) ; the IQ of 1080 children was 107.75 ± 16.81 ; median urinary iodine level of 358 children was 201.4 μg/L.②The median of salt iodine content was 28.6 mg/kg,the coverage rate of iodized salt was 98.8％ (1186/1200),and qualified rate of iodized salt was 93.0％ (1116/1200).③The residents average daily salt intake was 10.5 g.④Average score of the questionnaire survey of 1084 5th-grade students was 4.2 points.Average score of 961 housewives was 4.4 points.Conclusions Various technical indicators show that IDD is in a sustained elimination state in Henan Province.Strengthen health education,enhance public awareness of disease prevention is still the important work ahead.