OBJECTIVE To explore the dyna mic change of lncRNA expression during lung carcino-genesis induced by urethane.METHODS A total of 40 BALB/c mice received weekly ip injection of urethane 1 g·kg -1 for four continuous weeks,mice were euthanized at 12th week or 24th week after the first urethane treat ment,respectively.The RNA of lung tissues were isolated and used for microarray analysis.Based on the results of the microarray we selected lncRNA-AK017233 for additional qPCR analysis in individual mouse.RESULTS The incidence of lung cancer were 85% and 100% at 12th week and 24th week after the first ad ministration of urethane,respectively.The multiplicity and dia meter of lung tu mors in 24 weeks treated group were statistically significant fro m those in 12 weeks treated group (P<0.01 ),and pathological analysis showed that tu mors were classifiable as moderately differ-entiated adenocarcino ma.Total of 26 Down-regulated lncRNAs in which lncRNA-AK017233 stand for the most down-regulated lncRNA were identified by microcarray analysis.qPCR detected that the lncRNA-AK017233 was significantly altered by 0.33 ti mes in lung tu mors of urethane treated group at 12th week, co mpared to parallel lung tissues in urethane treated group at 12th week.AK017233 expression of ure-thane treated group was significantly reduced by 0.22 ti mes at 24th week,co mpared to parallel lung tis-sues in urethane treated group at 24th week.CONCLUSION LncRNA-AK017233 was consistently down-regulated during urethane induced lung carcinogenesis.

Objective To analyze the expression of long non-coding RNA ( lncRNA) in renal clear cell carcinoma ( RCCC ) , the association of lncRNA with RCCC, as well as the role of lncRNA in the diagnosis and treatment of RCCC.Methods Forty fresh RCCC tissues and their normal adjacent tissues were collected from March 2012 to June 2013, and total RNA was extracted using Trizol reagents, purified and tested by denaturing agarose gel electrophmesis and NanoDrop 1000.Through Arraystar Human LncRNA Microarray, the different expression of lncRNA between RCCC and normal adjacent tissues was screened. RT-qPCR was used to verify the expression of lncRNA in 40 pair RCCC tissues and normal adjacent tissues. The receiver-operating characteristic ( ROC ) curve was adopted to verify the diagnostic efficiency of the selected lncRNA.Results LncRNA expression profile showed 1 787 lncRNA with expression alteration in two fold or above, up-regulated and down-regulated candidate lncRNAs were 941 and 846 respectively. Compared with the adjacent tissues, NR_034095 and NR_038974 were up-regulated in RCCC, and ENST00000571724 and ENST00000566575 were down-regulated, which were consistent with the microarray analysis.By the ROC curves of NR_034095, NR_038974, ENST00000571724 and ENST00000566575 to discriminate the RCCC from normal adjacent tissue, the area under curve was 0.928 ( 95%CI 0.873 -0.984), 0.759 (95%CI 0.647-0.871), 0.833 (95%CI 0.747-0.919) and 0.887 (95%CI 0.815-0.959 ) , respectively.Conclusions NR _ 034095, NR _ 038974, ENST00000571724 and ENST00000566575 are significantly differently expressed in RCCC.The different expressed lncRNA might be closely related to the process of RCCC, and may be used as a new candidate target for molecular diagnosis and gene therapy of RCCC.

OBJECTIVE To prevent nosocomial infection and increase the safety of blood supply so as to evaluate the feasibility of routine detections in screening blood donations for HBV DNA and HBV markers. METHODS(Three hundred blood) donors and 346 cases were detected by ELISA for HBV markers,and then performed by LightCycler for HBV DNA. RESULTS One(1.6%) of 62 negative samples of HBV markers in 346 cases was positive for HBV DNA.Out of 300 qualified donors in screening tests,six(2%) were positive for HBV DNA.And two(0.9%) of 222 negative samples of HBV markers were positive for HBV DNA. CONCLUSIONS This study shows that there are HBV infections with negative hepatitis B surface antigen(HBsAg) in population of Qingdao area and that incorporating fluorescence quantitative PCR into ELISA in screening blood donations for HBV infection will further increase the safety of blood supply and prevent nosocomial infections.

Objective To explore the effect of mesenteric lymphatic ligation on liver injury induced by severe acute pancreatitis. Methods 54 SD rats are randomly divided into sham-operation group (A), severe acute pancreatitis group (B) and severe acute pancreatitis and ligation group (C). Liver tissues were collected from each rat in 6, 12, and 24 h after ligation. Pathological changes in liver tissues were observed by haematoxylin and eosin staining. Levels of serum glutamic-pyruvic transaminase and glutamic oxalacetic transaminase were detected. Levels of TNF-α and IL-1 in liver homogenate were examined by enzyme linked immunosorbent assay. Results Under light microscopic examination, neutrophils and inflammatory exudate in hepatic lobule was increased as time prolonged in group B. Inflammation was reduced in group C as compared with group B.Levels of serum glutamic-pyruvic transaminase and glutamic oxalacetic transaminase were more significantly increased in groups B and C than in group A (P<0.05);while they were more significantly decreased in group C than in group B. Levels of TNF-αand IL-1 in liver homogenate were significantly increased in group B as compared with group A (P < 0.05); whereas they were more significantly decreased in group C than in group B at 24 h (P < 0.05). Conclusions Mesenteric lymphatic ligation can reduce liver injury in severe acute pancreatitis. Mesenteric lymphatic fluids may play an important role in severe acute pancreatitis.

Objective To investigate antibiotics resistant characteristics and carbapenemases genotype of Acinetobacter baumannii in Intensive Care Unit (ICU),so as to provide theoretical basis for clinical prevention and treatment.Methods Retrospective study was made on 90 non-duplicated clinical isolates of Acinetobacter baumannii,which were collected From January 2013 to January 2014 in three tertiary hospitals of Qingdao.All strains were identified by VITEK2 automated microbiology analyzer;K-B method was used to do drug susceptibility test;polymerase chain reaction (PCR) was used to amplify the OXA-23,OXA-24,OXA-51,OXA-58,KPC-2,VIM,IMP genes,and the positive products of genes were sequenced;the chi-square test was used to compare the difference of the resistance rates.Results The detection rate of multi-drug resistant A.baumannii (MDRAB)and Pan-drug resistant A.baumannii (PDRAB)was 61.11% (55/90) and 17.78% (16/90).In the 32 strains of imipenem-resistant Acinetobacter baumannii,the resistant rates to Cefoperazone/sulbactam,Polymyxin B was lower,while the resistant rates to other drugs tested were more than 85%.The difference of the resistance rates to 9 drugs between imipenem resistant group and Imipenem sensitive group were statistically significant (P≤0.05).PCR result showed: 32 strains detected OXA-51 gene,28 strains detected OXA-23 gene,and 3 strains detected VIM gene,the detection rates of which were 100%,87.50% and 9.38% respectively.All strains were not detected OXA-24,OXA-58,KPC-2 and IMP genes.The sequenced results were absolutely homology with the corresponding genes in genbank.Conclusions The resistance of A.baumannii in ICU is serious in this region,especially imipenem-resistant A.baumannii,which were nearly no-sensitive to most of the drugs commonly used in clinical.The gene existence of carbapenemase and carbapenemase producing is one of the main resistance mechanism of Acinetobacter baumannii to carbapenem antibiotics.OXA-23 was the major genotypes in this region.

Objective:To investigate the effects of HLA-B?1502 genetic polymorphism on cyclosporine( CsA)-induced liver injury in Chinese renal transplant recipients. Methods:HLA-B?1502 genotypes were determined by polymerase amplification chaln reaction of sequence-specific primers( PCR-SSP) in a total of 339 renal transplant recipients receiving CsA. All the subjects were divided into the CsA-induced liver injury group, non-CsA-induced liver injury group and the control group according to the liver injury occurrence. Results:In the 339 renal transplant recipients, the frequency of HLA-B?1502 mutation allele was 22. 64%. The distribution frequen-cy of HLA-B?1502 mutation allele had no significant difference among the three groups. There were no significant differences in the clinical characteristics of HLA-B?1502 genotypes among three groups(P>0. 05). Conclusion: No association is observed between HLA-B?1502 genetic polymorphism and cyclosporine-induced liver injury in Chinese renal transplant recipients.

Objective To investigate the effect of interfering Hiwi gene on the apoptosis of MDA-MB-231 cells. Methods The mRNA and protein expression of Hiwi mRNA and its target protein were analyzed by qRT-PCR and Western Blot after transfection. MDA-MB-231 cells were divided into 6 groups according to the experimental design. Interference effects were screened as siRNA interference group (Hiwi10330 group), and then divided into 3 groups according to the experimental design: interference group, negative control group/NC, blank control group/Blank. The cell apoptosis rate was detected by flow cytometry after transfection. Results The expression of mRNA in the interference group was significantly lower than that in the siRNA group (P < 0.05), the expression of target protein of Hiwi gene was also significantly inhibited (P < 0.05). The apoptosis rate of MDA-MB-231 cells was significantly higher than that of NC and Blank groups (P<0.05). Conclusion The apoptosis rate of breast cancer cells MDA-MB-231 was significantly increased after siRNA targeting hiwi gene silencing.

Objective:To investigate the clinical effect of the transverse rectus abdominismuscle (TRAM) on reconstruction of the breast.Methods:The clinical data of 23 patients receiving TRAM breast reconstruction in our department from Jan. 2018 to Dec. 2019 were retrospectively analyzed.Results:The operation time of 23 patients ranged from 240 to 360 mins, andthe average time was about 300 mins. Intraoperative bleeding was about 120 to 200 ml, with an average of 170 ml. All the flaps survived successfully, but 2 cases were complicated with local fat necrosis. The postoperative period was between 6 and 12 months. No local tumor recurrence or metastasis was found inall patients during postoperative follow-up, and the breast shape was maintained in good condition.Conclusion:TRAM can make up for the regret of breast loss caused by breast cancer in female patients. It can bring confidence in life and work to female patients, and the technology is safe and reliable, which is worthy of promotion.

Objective To construct a nomogram prognostic model for predicting the survival of patients with lung adenocarcinoma based on the large sample data from the SEER database. Methods We retrospectively analyzed the clinical data of patients who were diagnosed with lung adenocarcinoma from 2010 to 2015 in the SEER database. A nomogram model was created based on independent parameters influencing the prognosis of patients with lung adenocarcinoma using Lasso Cox regression analysis. The C-index and calibration curve were utilized to assess the ability to distinguish and calibrate the nomogram. NRI and DCA curves were used to evaluate the prediction ability and net benefit of the nomogram. Results A total of 15 independent risk factors affecting the prognosis of lung adenocarcinoma were identified and integrated into the nomogram model. The C-index of the prediction model was 0.819 in the training cohort and 0.810 in the validation cohort. The predicted specific survival rate of the 1-, 3- and 5-year calibration curves of the training cohort and the validation cohort were consistent with the actual specific survival rate. In comparison to the 7th edition of the AJCC TNM staging system, the NRI and DCA curves demonstrated a considerable boost to the predictive capacity and net benefits achieved by the nomogram model. The risk stratification model constructed with this nomogram model was able to distinguish the patients with different risks well (P < 0.0001). Conclusion A nomogram prognostic model is successfully developed and validated, which provides a simple and reliable tool for the survival prediction of the patients with lung adenocarcinoma. Meanwhile, the risk stratification model constructed by the prediction model can conveniently screen patients with different risks, which is important for the individualized treatment of lung adenocarcinoma patients.

Objective:To explore the effect of signal transducer and activator of transcription 6 (STAT6) on ferroptosis in skeletal muscle cells in sepsis model and its potential mechanism.Methods:Twenty-four 8-week-old male specific pathogen free Kunming mice were divided into normal control group, sham group, sepsis model group and STAT6 inhibitor pretreatment group according to random number table method with 6 mice in each group. A mouse sepsis model was reproduced by cecal ligation and perforation (CLP). In the sham group, the skin of mice was sutured after exposing the cecum tissue. In the STAT6 inhibitor pretreatment group, 10 mg/kg AS1517499 was injected intraperitoneally 1 hour before model reproduction. The sham group and the model group were intraperitoneally injected with the same volume of normal saline. Mice in the normal control group did not receive any operation or drug intervention. The mice were sacrificed 24 hours after model reproduction, and the muscle tissue of hind limb was obtained under sterile condition. Hematoxylin-eosin (HE) staining was used to observe the histopathology with optical microscope, and mitochondrial morphological changes were observed by transmission electron microscopy after double staining with uranium acetate lead citrate. The ferroptosis marker proteins expressions of chitinase-3-like protein 1 (CHI3L1), cyclooxygenase-2 (COX-2), acyl-CoA synthetase long-chain family member 4 (ACSL4), ferritin heavy chain 1 (FTH1), and glutathione peroxidase 4 (GPx4) were detected by Western blotting.Results:Under the optical microscope, the morphology and structure of skeletal muscle tissues in the normal control and sham groups were normal. In the model group, the structure of skeletal muscle tissues was loose, the muscle fiber became smaller and atrophic, inflammatory cell infiltration and even muscle fiber loss were found. Compared with the model group, the structure of skeletal muscle tissues was tight and skeletal muscle atrophy was improved in the STAT6 inhibitor pretreatment group. The ultrastructure of skeletal muscle cell in the normal control and sham groups was normal under transmission electron microscope. The ultrastructure characteristics of skeletal muscle in the model group showed that cell membrane was broken and blister, mitochondria became smaller and membrane density increased, the mitochondrial crista decreased or disappeared, the mitochondrial outer membrane was broken, and the nucleus was normal in size but lacked chromatin condensation. Compared with the model group, the STAT6 inhibitor pretreatment group had a significant improvement in the ultrastructure of muscle cells. Compared with the normal control and sham groups, the protein expressions of CHI3L1, COX-2, ACSL4 and FTH1 in the muscle of the model group were significantly increased, while the protein expression of GPx4 was decreased significantly, indicating that the skeletal muscle cells in the mouse sepsis model showed characteristic mitochondrial injury and abnormal expression of ferroptosis markers. Compared with the model group, the protein expressions of CHI3LI, COX-2, ACSL4 and FTH1 in the STAT6 inhibitor pretreatment group were significantly decreased [CHI3L1 protein (CHI3L1/GAPDH): 0.70±0.08 vs. 0.97±0.09, COX-2 protein (COX-2/GAPDH): 0.61±0.03 vs. 0.83±0.03, ACSL4 protein (ACSL4/GAPDH): 0.75±0.04 vs. 1.02±0.16, FTH1 protein (FTH1/GAPDH): 0.49±0.06 vs. 0.76±0.13, all P < 0.05], while the protein expression of GPx4 was significantly increased (GPx4/GAPDH: 1.14±0.29 vs. 0.53±0.03, P < 0.05). Conclusions:Sepsis can induce ferroptosis in skeletal muscle cells of mice. STAT6 may mediate ferroptosis in mouse skeletal muscle cells by regulating the expressions of COX-2, ACSL4, FTH1 and GPx4, thereby inducing skeletal muscle cell injury in sepsis.