Objective To construct the eukaryotic expression vector of pcDNA3.1-PEA-15 and express it in the human esophageal cancer (EC-109) cells,and to explore the effect of PEA-15 on EC-109 cells.Methods The PEA-15 gene was amplified from EC-109 by reverse transcriptase polymerase chain reaction (RT-PCR) and ligated to eukaryotic expression vector pcDNA3.1.After confirmation of recombinant plasmid was correctly by endonuc]eases digestion and DNA sequencing,the construct was transfected it into EC-109 through liposome inducing.The expression of PEA-15 in transfected EC-109 was detected by RT-PCR and Western blot.The cell growth inhibition ratio was evaluated by MTT assay.Results RT-PCR indicated that PEA-15 was highly expressed in EC-109 cells.The amplified fragment by RT-PCR was coincident with hypothesis enzyme digestion analysis and DNA sequencing confirmed that the pcDNA3.1-PEA-15 was constructed successfully.The expression of PEA-15 gene was increased obviously in the transfected EC-109 detected by RT-PCR and Western blot respectively (t =4.078,5.269,P ＜ 0.05).The cell growth inhibition ratio in the group which transfected pcDNA3.1-PEA-15 was significantly lower compared with the pcDNA3.1 transfect group after 72 hours (t =6.163,P ＜ 0.05).Conclusions The recombinant eukaryotic expression vector pcDNA3.1-PEA-15 is constructed successfully,and it can be expressed in EC-109.It also shows that PEA-15 has the function on cell growth which suggests that PEA-15 can inhibit the apoptosis of EC-109 cells and its expression involved in esophageal cancer development.

Objective To evaluate the efficacy and safety of percutaneous nephrolithotomy (PCNL)guided by ultrasonography through upper pole access. Methods From October 2007 to October 2009, 42 patients with upper urinary tract calculi underwent PCNL through upper pole access.Among these cases, there were 10 cases of staghorn calculi, 22 cases of renal pelvis calculi, 7 cases of the upper calyx calculi, 3 cases of the lower calyx calculi, 4 cases combined with ureter calculi and 2 cases combined with ureteropelvic junction obstruction. The stone measured from 2.0 to 6.5 cm (average: 3.4 cm) in length. Working tunnels (F16-F26) were established through the 10th or llth intercostals. Pneumatic or holmium laser lithotripsy was used to disintegrate and remove stones by nephroscopy or ureteroscopy. Clinical data including operation time, complications and stone free rate were analyzed retrospectively. Results All the operations were completed in one session, single tract was used in 36 cases(85.7%), double tracts were used in the other 6 cases(14.3%). The stonefree rate after one session was 88.1% (37/42), 3 cases(7.1%) received a second-session PCNL, 2 cases (4.8%)underwent ESWL after operation. The mean operative time was 65 min(30- 140 min).Postoperative surgery-related infection rate was 9. 5% (4/42). One patient (2. 4%)required blood transfusion. Perforation of the pelvis occurred in 1 patient(2.4 %). No pleural or important organ injury occurred. Conclusion The upper pole access for PCNL can be convenient to remove stones,this method is a highly efficient and safe technique.

Objective To investigate the characteristics of IL-22 in Helicobacter pylori (H.pylori)infection.Methods Thirty H.pyloripositive and fifteen H.pylori negative gastric biopsy specimens were enrolled,IL-22 mRNA expression was detected by real-time PCR and the protein level of IL-22 was determined by ELISA in gastric tissue.The H.pyloriand cell coculture system was established and IL-22 expression was measured by real-time PCR to investigate the main source of IL-22 in gastric tissue.The IL-22-producing T cell was examined by FCM in the gastric mucosa tissue.Results Gastric mucosal IL-22 mRNA and protein levels were significantly higher in H.pylori-positive patients than uninfected patients (P＜0.05).A H.pyloriand cell coculture system was established successfully and gastric epithelial cell and T cell were the main source of IL-22 in gastric tissue.IL-22 was produced by CD4+ and CD8+ T cells and these T cells were increased in H.pylori-infected gastric mucosa (P＜0.05).Conclusion IL-22 took part in H.pyloriinfection induced immune response and increased IL-22-producing T cells was the important feature of H.pyloriinfection.

Apoptosis has been considered as the only form of regulated cell death for a long time. However, a novel form of programmed cell death called necroptosis was recently reported. The process of necroptosis is regulated and plays a critical role in the occurrence and development of multiple human diseases. Thus, the study on the molecular mechanism of necroptosis and its effective inhibitors has been an attractive field for researchers. Herein, we introduce the molecular mechanism of necroptosis and focus on the literature about necroptosis drug screening in recent years. In addition, the identification of the critical drug targets of the necroptosis is also discussed.

Objective To evaluate the effect of astragaloside Ⅳ on postoperative cognitive function in aged rats.Methods Sixty healthy male Sprague-Dawley rats,aged 22 months,weighing 360-480 g,were divided into 4 groups(n=15 each)using a random number table:control group(group C),surgery group(group S),low-dose astragaloside Ⅳ group(group L-AGS)and high-dose astragaloside IV group(group H-AGS).At 3 days prior to surgery,astragaloside Ⅳ 20 and 40 mg/kg were injected intraperitoneally once a day in L-AGS and H-AGS groups,respectively.The equal volume of normal saline was given instead in C and S groups.The animals underwent splenectomy under anesthesia with 1.8% isoflurane in S,L-AGS and H-AGS groups.Five rats in each group were randomly sacrificed at 1 day after operation,the hippocampi were removed for determination of interleukin-1beta(IL-1β),tumor necrosis factor-alpha(TNF-α)and IL-6 contents(by enzyme-linked immunosorbent assay)and expression of activated caspase-3,Bax and Bcl-2(by Western blot).The left animals underwent Morris water maze test at 15 days after operation.Results Compared with group C,the escape latency was significantly prolonged,the frequency of crossing the original platform was reduced,the space exploration time was shortened,the expression of activated caspase-3 and Bax was up-regulated,the expression of Bcl-2 was down-regulated,and the IL-1β,TNF-α and IL-6 contents were increased after operation in group S(P<0.05).Compared with group S,the escape latency was significantly shortened,the frequency of crossing the original platform was increased,the space exploration time was prolonged,the expression of activated caspase-3 and Bax was down-regulated,the expression of Bcl-2 was up-regulated,and the IL-1β,TNF-α and IL-6 contents were decreased after operation in L-AGS and H-AGS groups(P<0.05).Compared with L-AGS,the escape latency was significantly shortened,the frequency of crossing the original platform was increased,the space exploration time was prolonged,and the TNF-α contents were decreased after operation in group H-AGS(P<0.05).Conclusion Astragaloside Ⅳ can improve the postoperative cognitive function in a dose-dependent manner in aged rats.

Objeetive To study the differences of the biological characteristics and immune regulation function of adipose mesenchymal stem cells (AMSCs)from psoriasis patients and healthy people.Methods AMSCs were isolated and cultured from human psoriatic and healthy adipose tissue,the phenotypes and cell cycle of AMSCs taken from three generation were detected by flow eytometry.Alkaline phosphate enzyme staining and oil red o staining were used respectively to identify their adipogenic and osteogenic capacity.Next,the levels of inflammation antimicrobial proinflammatory factor were detected by PCR and ELISA.Then gene expression profile of AMSCs were screen by gene expression profile chip,as so to bolting the the gene array related with immunology gene.Results There was no significant change in cell morphology,and cell surface markers were expressed high for CD29,CD44,CD73,while lower for CD31,CD45 and HLA-DR.AMSCs of psoriasis patients and healthy people both had the ability of adipogenic and osteogenic differentiation.But the cell cycle showed the third generation AMSCs proliferation rates were slower than that of normal control,as compared with healthy controls,adipogenic differentiation ability was stronger.What'more,the level of inflammatory cytokines in psoriasis group was lower than that in controls such asIL-10,IDO,TGF-β,on the contrary the levels of proinflammatory factor in psoriasis group were higher than that in controls,such as TNF-α,IFN-γ.In addition,gene chip results suggested that psoriasis group AMSCs had obvious expression differences on JAK-STAT pathway with healthy controls.Conclusions Compared with the control,there are significant differences in patients AMSCs proliferation and adipogenic differentiation ability,immune inflammation suppression control ability is weaken,this phenomenon may be associated with JAK-STAT immune pathways related to downgrade.

Objective To analyze the concordance of KRAS, NRAS, BRAF and PIK3CA gene mutations detected in plasma and matched tumor tissues in colorectal cancer patients, in order to provide good evidences to support plasma could be a potential surrogate of tumor tissue for gene mutation test. Methods One hundred and seventy?five cases of colorectal cancer were collected at the First Hospital of Jilin University, from October 2016 to October 2017.There were 101 males and 74 females, their ages ranged from 28 to 85 years,with median age of 59 years. The KRAS, NRAS, BRAF and PIK3CA gene mutations in the plasma and paired tumor specimens of all patients were detected by next generation sequencing. Results The results of tissue samples test were gold standard. Comparison of the four genes showed that concordance rates between plasma and tissue samples were 81.1%(Kappa=0.543), 99.4%(Kappa=0.886), 99.4% (Kappa=0.886) and 97.7%(Kappa=0.714) respectively for KRAS, NRAS, BRAF and PIK3CA. The plasma detection rates of these genes were related to tumor stage(P=0.001), but not to gender(P=0.468) and age(P=1.000) of patients. Conclusions The study shows a high concordance of KRAS, NRAS, BRAF and PIK3CA gene mutations in plasma against mutation status in tumor tissue. In colorectal cancer, tumor tissue remains the best specimen for gene detection. However, patients from tumor tissue specimens cannot be obtained, especially those with advanced metastases, plasma can be used instead of tissue to detect the mutation status of KRAS, NRAS, BRAF and PIK3CA to guide targeted therapy.

Objective:To investigate the association of FAT atypical cadherin 1 (FAT1) with clinicopathological parameters and prognosis in esophageal squamous cell carcinoma (ESCC).Methods:The retrospective cohort study was conducted. The clinicopathological data of 124 patients with ESCC who were admitted to Shanxi Cancer Hospital from January 2011 to December 2015 were collected. There were 85 males and 39 females, aged from 40 to 72 years, with a median age of 60 years. The ESCC tissues surgically removed and adjacent tissues specimens were collected to prepare tissue microarray for immunohistochemical staining. The 5 cases of ESCC tissues and adjacent tissues were analyzed by real-time quantitative polymerase chain reaction (qRT-PCR). Observation indicators: (1) the expression of FAT1 protein in ESCC and adjacent tissues; (2) the expression of FAT1 RNA in ESCC and adjacent tissues; (3) the expression of FAT1 protein in ESCC tissues and its association with clinicopathological parameters; (4) follow-up and survival. Follow-up using outpatient examination and telephone interview was conducted to detect survival of patients up to February 13, 2019. The survival time was from surgical date to tumor-related death or endpoint of follow-up. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the t test. Measurement data with skewed distribution were represented as M (range). Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. Comparison of ordinal data was analyzed using the non parameter rank sum test. The Kaplan-Meier method was used to calculate survival time, and Log-rank test was used for survival analysis. Results:(1) The expression of FAT1 protein in ESCC and adjacent tissues: of 124 specimens, the 107 cases of ESCC tissues and 93 cases of adjacent tissues were finally obtained because of exfoliative tissues. There were 76 cases of ESCC tissues and corresponding adjacent tissues matched. Results of immuno-histochemical staining showed that FAT1 protein was expressed in both ESCC and adjacent tissues and was brown after staining. FAT1 was located in cytomembrane, with high expression of FAT1 as ≥75 and low expression as <75. The relative expression levels of FAT1 protein in ESCC and adjacent tissues were 68±42 and 77±37, showing a significant difference between ESCC and adjacent tissues ( t=2.380, P<0.05). (2) The expression of FAT1 RNA in ESCC and adjacent tissues: results of qRT-PCR showed that the relative expression levels of FAT1 RNA in 5 cases of ESCC and adjacent tissues were 1.6±0.4 and 2.5±0.3, with a significant difference between them ( t=3.560, P<0.05). (3) The expression of FAT1 protein in ESCC tissues and its association with clinicopathological parameters: of the 107 ESCC patients, 58 cases had high expression of FAT1. There were 42 and 16 cases with high expression of FAT1 in 65 non-drinking patients and 42 drinking patients, respectively, showing a significant difference between them ( χ2=7.229, P<0.05). (4) Follow-up and survival: 96 of 107 ESCC patients were followed up for 38.0?94.9 months, with a median follow-up time of 45.9 months. Survival analysis showed that the survival time of patients with high FAT1 expression was 24 months, versus 22 months of patients with low FAT1 expression, indicating no significant difference between them ( χ2=1.773, P>0.05). Results of subgroup analysis showed that the survival time was 24 months and 21 months of female patients with high and low FAT1 expression, 23 months and 22 months of non-smoking patients with high FAT1 expression and low FAT1 expression, 23 months and 21 months of non-drinking patients with high FAT1 expression and low FAT1 expression, respectively, showing significant differences between them ( χ2=8.769, 12.827, 10.724, P<0.05). Conclusions:The expression of FAT1 in ESCC tissues is low. Female, non-smoking and non-drinking ESCC patients with high FAT1 expression have good survival.

Objective: To investigate the intentions of mothers of ageappropriate girls in Qingdao to vaccinate their daughters against human papillomavirus (HPV), so as to provide theoretical guidance for targeted health education in the future. Methods: A multistage random sampling method was adopted to conduct a crosssectional study among 2 244 mothers of girls aged 12-14 years in Qingdao from March to December 2023. The Mann-Whitney U test was used for group comparisons, and Logistic regression was performed to analyze the factors that influenced maternal intention to vaccinate their ageappropriate daughters against HPV. Results: Among the surveyed mothers, 89.22% (n=2 002) intended to vaccinate their daughters against HPV, and 68.58% (n=1 539) had fully vaccinated or had plans to complete it for themselves. The knowledge score of mothers intended to vaccinate their daughters was 10 (8, 11). The multivariate Logistic regression analysis showed that mothers aged >45 years (OR=0.19), those with an annual family income of 60 000-<150 000 yuan (OR=0.65), 150 000-<300 000 yuan (OR=0.58), 300 000-500 000 yuan (OR=0.22), and those with higher knowledge scores (OR=0.90) were more likely to vaccinate their daughters (P<0.05). Mothers with a junior college or undergraduate degree (OR=1.66), those who never or occasionally screened for HPV (OR=1.58), those who were intended to be vaccinated, not planning to complete the fullcourse vaccination, or overaged and unvaccinated (OR=7.13), those who were not concerned about their daughters HPV infection (OR=2.54), and those whose daughters were not in adolescence (OR=1.93) were less intended to vaccinate their daughters (P<0.05). The primary reasons for vaccine hesitancy were vaccine safety concerns (65.06%), followed by the belief of mothers that "the children is to young, and can be vaccinated when they are older" (13.25%). Conclusions Mothers of eligible girls in Qingdao have relatively higher intentions to vaccinate their daughters against HPV, and willingness is influenced by factors such as the mothers vaccination status, knowledge level, and daughters development stage. It is recommended to strengthen targeted health education, improve the cognitive level and acceptance of mother, and increase the vaccination rate of HPV vaccines.

Objective:To investigate the expression of miRNA-34b (miR-34b) in non-small cell lung cancer (NSCLC) tissues and its effect on proliferation and invasion of human NSCLC A549 cells in vitro.Methods:The specimens of cancer tissues and paracancerous normal epithelial tissues (more than 5 cm from the edge of the tumor) were collected from 40 NSCLC patients in Shanxi Province Cancer Hospital from June 2015 to March 2017. A549 cells were transfected with miR-34b mimics (experimental group) and irrelevant sequences (negative control group), respectively. The expression of miR-34b in tissues and each group of A549 cells was detected by reverse transcription real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). The proliferation activity of A549 cells in the experimental group and the negative control group was detected by methyl thiazolyl tetrazolium (MTT) assay, and the invasion ability of A549 cells in the two groups was detected by Transwell assay.Results:The relative expression of miR-34b in NSCLC tissues was lower than that in paracancerous normal epithelial tissues (0.52±0.06 vs. 1.05±0.17), and the difference was statistically significant ( P < 0.001). The relative expression of miR-34b in A549 cells of the experimental group was higher than that in the negative control group, and the difference was statistically significant ( P < 0.05). MTT assay showed that the cell proliferation ability (absorbance value) of A549 cells in the experimental group was lower than that in the negative control group after cultured for 24 and 48 hours (both P < 0.01). Transwell assay showed that the number of invaded A549 cells in the experimental group was less than that in the negative control group [(49.53±5.03) cells vs. (121.00±12.06) cells, P < 0.01]. Conclusions:The expression of miR-34b is low in NSCLC tissues, and the up-regulation of miR-34b expression can inhibit the proliferation and invasion of NSCLC A549 cells.