Objective To evaluate the impact of small interference RNA (siRNA)-induced silencing of adrenomedullin (ADM) gene on the growth of and apoptosis in a malignant melanoma cell line A375.Methods Three siRNAs targeting ADM gene were constructed and transfected into A375 cells.Real-time quantitative reverse transcription PCR (qRT-PCR) was performed to measure the expression of ADM to select the most efficient siRNA for the next experiment.A375 cells were classified into 3 groups to be transfected with the selected siRNA (test group),non-specific sequences (negative control group),or remain untransfected (blank control group).Then,an immunohistochemical SP method was used to determine the expression of ADM at 24hours,methyl thiazolyl tetrazolium (MTT) assay to detect the proliferation of A375 cells at 24 and 48 hours,flow cytometry to observe cell apoptosis at 48 hours.Results qRT-PCR showed that ADM-siRNA-2 was the most efficient.The expression of ADM was significantly lower in the test group than in the other two groups.The proliferation level (represented as the absorbance value at 490 nm) of A375 cells was 0.389 ± 0.0375 and 0.469 ± 0.0330 in the test group at 24 and 48 hours respectively,significantly lower than that in the negative control group (0.548 ± 0.0376,0.676 ± 0.0374,both P＜ 0.05) and blank control group (0.574 ± 0.0733,0.685 ± 0.0154,both P ＜ 0.05).Flow cytometry revealed a statistical increase in early apoptosis rate in the test group compared with the blank control group and negative control group ((20.200 ± 6.046)％ vs.(1.667 ± 0.340)％ and (4.587 ± 1.294)％,both P ＜ 0.05).No significant differences were observed in the proliferation level or early apoptosis rate between the negative control group and blank control group (both P ＜ 0.05).Conclusion The siRNA-induced silencing of ADM gene can inhibit the proliferation of,but promote the apoptosis in,A375 cells.

Objective To measure the level of circulating endothelial progenitor cell (EPC) and serum vascular endothelial growth factor (VEGF), and to investigate its clinical significance in patients with different stages of renal cell carcinoma (RCC). Methods The level of circulating EPC was quantified by assaying CD45-CD34+VEGFR-R2 +cell phenotype in 45 patients with RCC (RCC group), 30 patients with benign renal tumors(benign renal tumors group) and 30 healthy controls (control group). Serum VEGF was quantified by enzyme- linked immunosorbent assay (ELISA). The results were compared. Results The level of EPC in RCC groups was (0.265 ± 0.042)%, in benign renal tumors group was(0.053 ± 0.008)% , and in control group was (0.048 ± 0.006)%. The level of EPC in RCC group was significantly higher than that in benign renal tumors group and control group (P<0.05). The level of EPC in Ⅲ- Ⅳ stage patients was significantly higher than that in Ⅰ-Ⅱ stage patients:(0.312 ± 0.038)%vs. (0.215 ± 0.021)%, P<0.05. Three months after operation, the level of EPC in 33 RCC patients without pretreatment was (0.078 ± 0.003)% and significantly lower that before treatment (P<0.05). The level of VEGF in RCC groups was (305.5 ± 29.1) ng/L, in benign renal tumors group was (29.8 ± 3.2) ng/L, and in control group was (25.1 ± 2.8) ng/L. The level of VEGF in RCC group was significantly higher than that in benign renal tumors group and control group (P<0.05). The level of VEGF inⅢ-Ⅳstage patients was significantly higher than that inⅠ-Ⅱstage patients:(365.6 ± 34.6) ng/L vs. (256.2 ± 23.2) ng/L, P<0.05. Pearson association analysis showed that the level of EPC had positive associations with VEGF (r=0.714, P<0.01). Multiple linear regression showed that the size of kidney neoplasms was a dependent factor for the level of EPC. Conclusions The level of EPC has a positive association with VEGF. EPC maybe a new biomarker for RCC.

Objective To establish the method of identifying MRSA with Taqman-fluorescence quantitative PCR basing on mecA/nuc/fem B three gene combined detecting.Methods Taking the coagulase positive MRSA,which isolated from the clinical samples and confirmed by VITEK 2 compact microbial analyzer,as the research obj ect,designed mecA/nuc/fem B specific PCR primers and Taqman fluorescent probe by bio-software PrimerPremier 5 and Designer Beacon 7,FAM,HEX and ROX markers were used to label the fluorescent probe at 5’,and the end of 3’was labeled with BHQ1,detected by fluo-rescence quantitative PCR instrment.Results ①1 g/dl gel electrophoresis results showed that the primer’s specificity of mec A/nuc/fem B were good,and molecular weight of the amplification band consistent with the expected molecular weight and no non-specific amplification band.②Three genes were obtained specific amplification in a single tube single channel and single tube multiple channel detection in PCR,and the three gene amplification effect in a single tube single tube single chan-nel and multichannel PCR similar.Conclusion Successfully established a method of multi channel Taqman-probe fluores-cence quantitative PCR identification of MRSA,mec A/nuc/fem B combined detection can effectively differentiate coagulase negative and positive MRSA,improve the accuracy of identification.

Objective To survey the prevalence of elderly asthmatics among urban and rural residents of Liaoning Province and to provide data for preventive and therapeutic policies of asthma. Methods Stratified cluster disproportional 2.5‰ random sampling survey for 116 276 re sidents was performed using uniform scheme, procedures and questionnaire. Among them 12 735 cases were over 60 years old. Results Totally 522 cases (207 male and 315 female) were diagnosed as asthma, the overa ll prevalence was 1 25% and that of elderly was 4 09% ( male 3 26%, female 4 92%, P

Objective To investigate the surgical options for the management of portal vein thrombosis (PVT) during liver transplantation and its impact on the outcome of patients. Methods 773 cases of liver transplantation were analyzed retrospectively. PVT occurred in 107 patients, inclu-ding 59 of grade Ⅰ ,33 of grade Ⅱ, 12 of grade Ⅲ and 3 of grade Ⅳ. Simple thrombectomy or thrombus-extraction was performed in grade Ⅰ and Ⅱ. 12 patients with grade Ⅲ received thrombus-extraction or using the donor iliac vein to act as a bridge between the donor portal vein and host superior mesenteric vein. Two cases of grade Ⅳ received a modified cavo-portal hemitransposition and one case received portal-vena coronaria varication anastomosis. Results Liver function had a good recover and the perio-perative mortality is 4. 3% in grade Ⅰ and Ⅱ. In grade Ⅲ , 5 cases received thrombus-extraction had a normal liver function after transplantation and had no died. 2 cases among the other 7 cases using por-tal vein reconstruction had bad liver function and died. The liver function recovered well after trans-plantation and there was no died in grade Ⅳ. Conclusions PVT is not a contraindication for liver transplantation. Good results can be obtained by applying reasonable operative procedures individually.

Objective To explore the variation trend of natural killer (NK) cell subsets in the recipients infected with cytomegalovirus (CMV) after renal transplantation. Methods Clinical data of 92 renal transplant recipients were retrospectively analyzed. All recipients were divided into the CMV infection group (n=43), CMV infection recovery group (n=13), stable renal function group (n=15), rejection group (n=11) and other infection group (n=10). In addition, healthy adult volunteers were enrolled in the healthy control group (n=15). The proportion of NK cells in peripheral blood, the expression proportion and the mean fluorescence intensity (MFI) of CD226 and CD16 in NK cells were observed and statistically compared among different groups. Results The proportion of NK cells was 4.9% (2.2%, 11.5%) in the CMV infection group and 3.7% (2.3%, 6.5%) in the CMV infection recovery group, which were significantly lower than those in the other groups (all P < 0.05). The expression proportion of CD226 and CD16 in NK cells in the CMV infection group was significantly lower compared with those in the healthy control group and stable renal function group(all P < 0.05). The expression proportion of CD226 and CD16 in NK cells in the CMV infection recovery group was remarkably higher than those in the CMV infection group (both P < 0.05). The MFI of CD226 and CD16 in the CMV infection group was significantly lower than those in the healthy control group (both P < 0.05). The MFI of CD226 and CD16 in the CMV infection recovery group was significantly higher than those in the CMV infection group (both P < 0.05). Conclusions The expression proportion and MFI of CD226 and CD16 in NK cells are down-regulated in CMV infection period, whereas up-regulated during the CMV infection recovery period, prompting that CD226 and CD16 expressed by NK cells are intimately correlated with the course of CMV infection.

Objective:To preliminarily observe the clinical efficacy of microwave hyperthermia combined with intensity-modulated radiotherapy (IMRT) and chemotherapy for patients with locally advanced gastric cancer.Methods:Forty patients who could not been operated or refused operation were enrolled in this clinical trial, who were confirmed as locally advanced proximal or distal gastric cancer by gastroscopy pathology and imaging. Radiotherapy was delivered by IMRT technology for 5 times per week with a total dose of 46 to 56 Gy (median dose of 50 Gy) in 25 to 28 fractions. Synchronous hyperthermia was given at 42 to 44℃ twice a week, 45 min/time. S-1 or capecitabine-based synchronous chemotherapy was performed, d1-14/3 weeks. The symptom remission rate, adverse reactions, objective remission rate (complete and partial remission) and survival were observed.Results:A total of 40 patients, aged between 56 and 83 years (median age of 71 years), were enrolled in this study. The male-to-female ratio was 7: 1. Among them, 38 cases (95%) showed symptom remission. The most common adverse reactions were grade 1-2 gastrointestinal reactions and leukopenia. The objective remission rate was 87.5%, the 2-year progression-free survival and overall survival rates were 68.6% and 70.5%, respectively.Conclusion:Preliminary findings demonstrate that microwave hyperthermia combined with chemoradiotherapy achieve satisfactory outcomes and yield tolerable toxicity in patients with locally advanced gastric cancer.

One patient with chronic refractory wound in right lower leg was admitted to our department in June 2017, which experienced repeated ulceration for 4 years. On 4 days post admission, two bamboo sticks were taken out from the deep muscle of the right lower leg. The wound was repaired by perforator flap of posterior tibial artery in the right lower leg. The donor site was covered with split-thickness skin in the upper leg of the same side. On 7 days post operation, the flap and skin graft survived. During follow-up of 12 months, the flap had good appearance. This case suggests that the first thing for the treatment of chronic refractory wound is to find out the cause of wound.

The clinical data of 67 patients underwent orthotopic liver transplantation, including 4 cases of early intra abdominal hemorrhage after orthotopic liver transplantation, were analyzed retrospectively.Anastomotic liver artery hemorrhage was found in 2 cases, hemorrhage of IVC in 1 case and hemorrhage of right adrenal gland in 1 case. All the 4 patients were correctly dignosed and successfully treated by operation in time. It suggests that the early intra abdominal hemorrhage after orthotopic liver transplantation is mostly due to surgical technique. The improvement of surgical technique,correct diagnosis and timely operation are very important for the treatment of intra abdominal hemorrhage.

Objective:To investigate the expression of forkhead box protein O3(FOXO3) in pancreatic cancer and its effect on the motility and proliferation of pancreatic cancer cells.Methods:The FOXO3 expression in pancreatic cancer and adjacent tissues was retrieved from LinkedOmics database. Western blotting and real-time quantitative polymerase chain reaction were used to detect FOXO3 expression in pancreatic cancer cells and human pancreatic stellate cells. PANC-1 and MIAPaCa-2 pancreatic cancer cells with low FOXO3 expression were selected to transfect FOXO3 overexpression plasmid and negative control plasmid, respectively. The motility and proliferation ability of pancreatic cancer cells were detected by colony formation assay, cell scratch assay, Transwell assay and flow cytometry.Results:In the LinkedOmics database, the relative expression of FOXO3 protein in the cancer tissues of 64 patients with pancreatic cancer was significantly lower than that in the adjacent tissues ( t=8.36, P<0.001). The number of clones in PANC-1 cell line was (30.0±6.6) after overexpressed FOXO3, which was lower than that in negative control cells (92.7±6.7), and the difference was statistically significant ( t=11.54, P<0.001). After overexpressed FOXO3 in PANC-1 and MIAPaCa-2 cell lines, the scratch repair rate was significantly decreased compared with the control group. In Transwell experiment, the number of cells in FOXO3 overexpressed group in PANC-1 cell lines was (21.0±6.6), which was lower than that of negative control groups (55.7±8.5), and the difference was statistically significant ( t=5.59, P=0.005). The results of MIAPaCa-2 cell line were consistent with that of PANC-1 cell line. After overexpressing of FOXO3 in PANC-1 and MIAPaCa-2 cell lines, the proportion of cells in the G0/G1 phase decreased, while the proportion in the S phase increased. Conclusion:The expression of FOXO3 was decreased in pancreatic cancer. Overexpression of FOXO3 could significantly inhibit the proliferation, migration and invasion of pancreatic cancer cells and induce cell cycle arrest, which is a potential target for the treatment of pancreatic cancer.