Objective To study the relationship between the methylation of p16 promotor and the pathogenesis and progress of psoriasis. Method Methylation-specific PCR and DNA sequencing were used to detect the methylation of p16 promotor. Results The methylation of p16 promotor was found in 23.08% (6/26) of psoriatic lesions and 19.23% (5/26) of non-lesional areas in psoriatic patients, but none in normal controls. The frequency of methylation of p16 promotor was higher in psoriatic lesions than that in normal skin (P

Objective: To observe the clinical effect of FU's subcutaneous needling on acute lumbar sprain. Method: One hundred acute lumbar sprain cases were randomly allocated into a treatment group and control group, 50 cases in each. FU's subcutaneous needling on tenderness were employed in the treatment group, whereas voltaren was administered to the cases in the control group. The changes in symptoms and signs were then observed in the two groups. Results: The total effective rates in the treatment group and control group were 94.0% and 70.0% respectively, showing a statistical difference (P<0.01). Conclusion: FU's subcutaneous needling is better than voltaren for acute lumbar sprain.

Objective To observe the effect of Radix astragali (RA) on myocardial connexin-43 in rats with dilated cardiomyopathy (DCM). Methods The dilated cardiomyopathy model in rat was established through intraperitoneal injection with adriamycin. The rats in the model group were randomly divided into RA group and the model control group according to different methods of administration. Rats in RA group were gavaged with Astragalus particles and double-distilled water, and rats in model control group and normal control group were gavaged with an equal amount of double-distilled water daily for four weeks. At the end of 12 weeks, the left ventricular ejection fraction (LVEF) was measured with echocardiogram. The Cx43 mRNA level was tested by reverse transcriptase polymerase chain reaction (RT-PCR). Immunohistochemical method was used to observe myocardial Cx43 expression and distribution. Results Compared with the model control group, the Cx43 mRNA and protein expressions and LVEF were increased signiifcantly in RA group (P<0.05). The disorders in distribution of myocardial Cx43 improved in RA group in contrast to the model control group. Conclusions Radix astragali can improve myocardial Cx43 expression and distribution in DCM rats, and can further improve the cardiac function.

Objective To evaluate the clinical value of midfrontal keyhole approach for the treatment of severe intra-ventricular hematoma. Methods The clinical data of 21 cases of severe intraventricular hemorrhage through midfrontal key-hole approach were analyzed retrospectively. Results Both inside and outside intraventricular hematoma were satisfied cleared. The GCS score and intraventricular hemorrhage Graeb score were improved. There were complications after opera-tion including 1 patient with diffuse brain swelling, 3 patients with cerebral vasospasm, 1 patient with intracranial infection, and seven patients with pulmonary infection. Follow-up schedules included 1-6 months. According to ADL score, 5 patients recovered well, 9 patients were moderately disabled, 3 were severely disabled, 1 was in a vegetative state and 3 died. Conclu-sion The intraventricular hematoma can be removed through midfrontal keyhole approach. The obstructive hydrocephalus can be relieved, the secondary brain damage was reduced and the prognosis was improved in patients.

Objective To investigate the risk factors to acute lung injury based on multiple trauma to provide a theoretical basis for early intervention .Methods The emergency surgical patients with multiple trauma in our hospital from March 2006 to March 2011 were selected .The patients meeting the diagnostic criteria for acute lung injury were taken as the study group and the others as the control group .All patients were enrolled for evaluating the injury severity score (ISS) ,acute physiology and chronic health Ⅱ(APACHE Ⅱ) score and recording smoking ,alcohol abuse ,diabetes mellitus ,number of organ damage ,gastrointestinal bleeding , pulmonary contusion ,diffuse intravascular coagulation (DIC ) ,vomiting ,traumatic shock ,time to correct shock ,blood transfusion . The polymorphism of rs3788853 ,rs13306087 ,rs12709426 of angiotensin-converting enzyme(ACE) gene were analyzed .Results In the study group and the control group ,there were statistical differences in 6 influencing factors of the ISS ,APACHE Ⅱ score ,blood transfusion ,DIC ,traumatic shock ,time to correct shock>6 h(P<0 .05);the gene and genotype frequencies of ACE between the two groups had no statistical difference (P>0 .05);the 6 kinds of influencing factors were risk to acute lung injury based on multi-ple trauma by Logistic regression analysis .Conclusion The ISS ,APACHE Ⅱ score ,blood transfusion ,DIC ,traumatic shock ,long time to correct shock are the risk factors to acute lung injury based on multiple trauma .

Objective To study the effect of down-regulation of PTTG on the proliferation and migration of cutaneous squamous cell carcinoma cell line SCL-1 and its related mechanism. Methods SCL-1 cells were transfected with control siRNA or PTTG-targeting siRNA (PTTG-siRNA), or remained untransfected. After additional culture, the proliferation of SCL-1 cells as observed with cell counting kit-8 (CCK-8), and cell migration with Boyden chamber. Real-time PCR and Western blot were performed to detect the expression of matrix metalloproteinase 2 (MMP-2), MMP-9 and PTTG. Results The proliferation of SCL-1 cells transfected with PTTG-siRNA was markedly deccelarated in comparision with that of untransfected cells and those transfected with control siRNA (both P< 0.05). Real-time PCR and Western blot disclosed a significant decrease in the mRNA and protein expression of MMP-2, MMP-9 and PTTG in PTTG-siRNA-transfected SCL-1 cells compared with the other two groups of cells. As real-time PCR showed, the expressions of MMP-2, MMP-9 and PTTG in PTTG-siRNA-transfected SCL-1 cells were 0.8%, 23.2% and 21.3% of those in untransfected cells, respectively. Further more, the number of SCL-1 cells migrating through microporous membrane in the Boyden chamber was significantly smaller in PTTG-siRNA-transfected group than in untransfected group and control siRNA-trans-fected group (51.38 ± 4.71 vs 131.33 ± 6.12 and 127.72 ± 5.20, both P< 0.05). Conclusion The down-regulation of PTTG may deccelarate the proliferation and migration of SCL-1 cells and inhibit the expression of MMP-2 and MMP-9 in SCL-1 cells.

Objective To evaluate effects of downregulation of glucose?6?phosphate dehydrogenase(G6PD) expression on proliferation and cell cycle distribution of cutaneous squamous cell carcinoma(CSCC)cells. Methods Western blot analysis was performed to measure the protein expression of G6PD in normally cultured human HaCaT keratinocytes, SCL?1 and A431 CSCC cells. When A431 cells grew to 85%-90%confluence, a small interfering RNA (siRNA)targeting G6PD(G6PD?siRNA group)and a negative control siRNA(siRNA control group)were transfected into them separately, and untransfected A431 cells served as the untransfected group. CCK?8 assay was performed to evaluate proliferative activity of the A431 cells on days 0, 1, 2, 3 and 4 after transfection, Western blot analysis to measure G6PD, cyclin D1 and CDK4 protein expressions in A431 cells, and flow cytometry to analyze cell cycle distribution in A431 cells after 48 hours of additional culture. Results The protein expression of G6PD was significantly higher in normally cultured SCL?1 cells(0.308 ± 0.023)and A431 cells(0.643 ± 0.046)than in HaCaT cells(0.100 ± 0.019, both P<0.05), and significantly higher in A431 cells than in SCL?1 cells(P<0.05). The G6PD?siRNA group showed significantly decreased protein expressions of G6PD, cyclin D1 and CDK4(0.134 ± 0.027, 0.154 ± 0.017 and 0.166 ± 0.017, respectively)compared with the untransfected group(0.425 ± 0.029, 0.344 ± 0.024 and 0.330 ± 0.020 respectively)and siRNA control group(0.444 ± 0.033, 0.350 ± 0.027 and 0.348 ± 0.018 respectively) (all P<0.05). Besides, the G6PD?siRNA group showed significantly decreased cellular proliferative activity on days 1-4 compared with the siRNA control group and untransfected group(all P<0.001), while there were no significant differences between the untransfected group and siRNA control group at any of the time points (all P > 0.05). Compared with the untransfected group and siRNA control group, the G6PD?siRNA group showed significantly higher proportions of A431 cells in G0/G1 phase(both P < 0.001), but significantly lower proportions of A431 cells in S phase(both P<0.001). Conclusion G6PD may play important roles in the regulation of proliferation and cell cycle distribution of CSCC cells.

ObjectiveTo investigate the role of Notch1 gene in xenografted human cutaneous squamous cell (SCL-1) carcinoma. MethodsFifteen nude mice were divided into three groups, including untreated group(inoculated with SCL-1 cells treated with phosphate buffered saline), empty vector group (inoculated with SCL-1 cells transfected with empty vector) and Notch1 group(inoculated with SCL-1 cells transfected with Notch1 expression vector). All the mice were inoculated with SCL-1 cells(1 x 108/ml) of0.2 ml. Then, the growth of xenografted tumor was observed every other day. Fifteen days later, the mice were sacrificed, tumor tissue was dissected and subjected to terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay for the detection of cell apoptosis, reverse-transcription(RT)-PCR and Western blot for the examination of mRNA and protein expressions of Notch1, bcl-2 and bax, respectively. ResultsThe proliferation of xenografted tumor in Notch1 group was obviously inhibited compared with the untreated group. The weight of xenografted tumor in Notch1 group was significantly lower than that in the untreated group and empty vector group (0.574 ± 0.219 g vs. 2.642 ± 0.404 g and 2.606 ± 0.512 g, F= 26.642, P＜ 0.01). TUNEL assay demonstrated that the number of apoptotic cells per 500 cells in tumor tissue specimens was(87 ± 9) in Notch1 group, evidently higher than that in the untreated group(8 ± 2) and empty vector group(10 ± 3) (F = 194.266, P ＜ 0.05 ). Further, RT-PCR and Western blot revealed that the mRNA and protein expressions of Notch1 and bax were significantly upregulated, but those of bcl-2 were markedly downregulated in the Notch 1 group, with significant difference among the three groups(all P ＜ 0.05). ConclusionsNotch 1 gene can inhibit the growth of xenogra ffted human cutaneous squamous cell(SCL-1) carcinoma and induce SCL-1 cell apoptosis likely by upregulating bax expression and downregulating bcl-2 expression.

Objective To evaluate the effects of shRNA-TGF-β1 plasmid on Smads signal transduction of rat renal allograft.Methods A Sprague-Dawley to Wistar rat orthotopic transplant kidney-sclerosis accelerated model was constructed and transfected with short hairpin RNA-TGF-β1 based on the hydromechanics.The recipients were divided into three groups:group T(plasmid group)injected with shRNA-TGF-β1;group H(vacant plasmid group)injected with vacant plasmid;group Y(simply transplantation group)injected with no plasmid.In group J(sham-operated group)only right kidney was removed with no transplantation as control group.Transplanted kidneys and blood samples were collected at the first,second and third month after transplantation.The blood urea nitrogen(BUN)and serum Cr were tested by enzyme-linked immunoadsordent assay.The gene transcriptional level of TGF-β1 and Smad3/7 was detected by RT-PCR,and the protein variations of TGF-β1 and phosphorylated Smad3/7 were examined by Western blotting.Results At each test time point,the BUN and serum Cr were significantly higher in the plasmid group than in the sham-operated group(P<0.05 or P<0.01),but obviously lower than in the vacant plasmid group and simply transplantation group(P<0.05 or P<0.01).The expression of TGF-β1 as well as phosphorylated Smad3 was significantly higher in the plasmid group than in the sham-operated group(P<0.05 or P<0.01),but obviously lower than in the vacant plasmid group and simply transplantation group(P<0.05 or P<0.01).However,the expression of phosphorylated Smad7 was significantly lower in the plasmid group than in the sham-operated group(P<0.05 or P<0.01),but obviously higher than in the vacant plasmid group and simply transplantation group(P<0.05 or P<0.01).Conclusion Short hairpin RNA-TGF-β1 plasmid could significantly improve the renal function of rat renal allografts probably by downregulating phosphorylated Smad3 and upregulating phosphorylated Smad7,leading to the inhibition of TGF-beta 1 promoting fibrosis role and delay of the allograft fibrosis.

Apoptosis ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Down-Regulation ; Glucosephosphate Dehydrogenase ; metabolism ; Humans ; Neoplasm Invasiveness ; Skin Neoplasms ; metabolism ; pathology