Objective:To investigate the influencing factors for synchronous colorectal liver metastasis (synCRLM).Methods:The retrospective case-control study was conducted. The clinicopathological data of 3 172 patients with primary colorectal cancer (CRC) who were admitted to the Affiliated Hospital of Qingdao University from January 2010 to January 2016 were collected. There were 1 946 males and 1 226 females, aged (63±12)years, with a range from 21 to 97 years. Observation indicators: (1) general data analysis; (2) clinicopathological data analysis; (3) analysis of influencing factors for synCRLM. Measurement data with normal distribution were represented as Mean±SD. Count data were represented as absolute numbers. The influencing factors for synCRLM were analyzed after excluding missing data of tumor differentiation degree, tumor diameter, pathological T stage and N stage. Univariate analysis was conducted by chi-square test or Logistic regression model. Multivariate analysis was conducted by Logistic regression model. Results:(1) General data analysis: among the 3 172 patients, cases with age ≤29 years, from 30 to 39 years, from 40 to 49 years, from 50 to 59 years, from 60 to 69 years, from 70 to 79 years, and ≥80 years were 15, 82, 342, 774, 965, 759 and 235, respectively. There were 2 972 patients in Qingdao, 172 cases in Yantai and 28 cases in Weihai. Of the 2 972 patients in Qingdao, there were 422 cases in Shinan District, 658 cases in Shibei District, 457 cases in Huangdao District, 144 cases in Laoshan District, 188 cases in Licang District, 205 cases in Chengyang District, 252 cases in Jimo District, 221 cases in Jiaozhou City, 255 cases in Pingdu City, 170 cases in Laixi City. (2) Clinico-pathological data analysis: among the 3 172 patients, there were 1 639 cases of colon cancer including 972 cases with left colon cancer and 667 cases with right colon cancer, 1 533 cases of rectal cancer. There were 2 981 cases of adenocarcinoma, 165 cases of mucinous adenocarcinoma, 10 cases of signet ring cell carcinoma and 16 cases of other types including carcinoid tumor, squamous carcinoma, tubular adenocarcinoma, etc.There were 162 cases with highly differentiated adenocarcinoma, 5 cases with highly-moderately differentiated adenocarcinoma, 2 338 cases with moderately differentiated adenocarcinoma, 80 cases with moderately-poorly differentiated adeno-carcinoma, 396 cases with poorly differentiated adenocarcinoma and 191 cases missing tumor differentiation data. There were 708 cases with tumor diameter <4 cm, 1 957 cases with tumor diameter ≥4 cm and 507 cases missing tumor diameter data. There were 486 cases in T1 or T2 stage of pathological T stage, 2 169 cases in T3 or T4 stage of pathological T stage and 517 cases missing tumor pathological T staging data. There were 1 563 cases in N0 stage of pathological N staging, 1 062 cases in N1 or N2 stage of pathological N staging and 547 cases missing tumor pathological N staging data. There were 2 895 cases without synCRLM and 277 cases with synCRLM. There were 2 799 cases without diabetes and 373 cases with diabetes. There were 2 931 cases without fatty liver and 241 cases with fatty liver. There were 2 989 cases negative for hepatitis B surface antigen (HBsAg) and 183 cases positive for HBsAg. (3) Analysis of influencing factors for synCRLM. Results of univariate analysis showed that gender, tumor location, tumor differentiation degree, tumor diameter, pathological T stage, fatty liver, HBsAg were related factors for synCRLM in primary colorectal cancer ( χ2=7.400, 7.577, 7.111, 4.513, 12.125, 5.686, 5.919, P<0.05), and neutrophils counts, lymphocytes counts, platelet counts, alanine aminotransferase (ALT), aspartate aminotrans-ferase (AST), total bilirubin, γ-glutamyltransferase (GGT), triacylglycerol (TG), total cholesterol (TC), carcinoembryonic antigen (CEA), and CA19-9 were related factors for synCRLM in primary colorectal cancer ( odds ratio=1.101, 0.807, 1.002, 1.017, 1.023, 1.027, 1.012, 0.686, 1.169, 1.007, 1.004, 95% confidence interval as 1.048-1.156, 0.678-0.960, 1.001-1.004, 1.011-1.024, 1.016-1.031, 1.011-1.044, 1.009-1.015, 0.541-0.869, 1.047-1.306, 1.006-1.008, 1.003-1.004, P<0.05). Results of multivariate analysis showed that cases as male, case with positive HBsAg, AST, GGT, TC, CEA and CA19-9 were independent risk factors for synCRLM in primary colorectal cancer ( odds ratio=1.503, 2.492, 1.018, 1.007, 1.301, 1.005, 1.003, 95% confidence interval as 1.038-2.178, 1.443-4.304, 1.003-1.034, 1.003-1.011, 1.112-1.522, 1.003-1.006, 1.002-1.003, P<0.05), and lymphocytes, ALT and TG were independent protective factors for synCRLM in primary colorectal cancer ( odds ratio=0.777, 0.983, 0.602, 95% confidence interval as 0.608-0.993, 0.966-0.999, 0.421-0.862, P<0.05). Conclusion:Cases as male, case with posotive HBsAg, AST, GGT, TC, CEA and CA19-9 are independent risk factors for synCRLM in primary colorectal cancer, while lymphocytes, ALT and TG are independent protective factors for synCRLM in primary colorectal cancer.

Objective:To construct RNA interference (RNAi) lentiviral expression vector of DEK gene, and to explore its effect on the biological behavior of liver cancer cells.Methods:Double-stranded oligo DNAs were annealed and synthesized according to the interference sequence of DEK gene by RNAi technology. Small interfering RNA expression vector pLKO.1 was cloned after enzymatic digestion. The recombinant lentiviral pLKO.1-sh hDEK was constructed, and then the virus supernatant was collected, packed and infected by 293T cells. Real-time reverse transcription PCR (RT-PCR) and Western blot were used to detect DEK expression in human liver cancer cells Bel-7402, Hu-7, SmMC-7721 and HepG2, and DEK knockdown efficiency in each group of lentivirus-infected cells. Cell proliferation ability, cloning ability, apoptosis and migration ability were detected by cell counting kit-8 (CCK8), flow cytometry and scratch test, respectively. The t-test was used to compare the mean between the two groups, and one-way ANOVA was used to compare the multiple groups.Results:Enzymatic digestion and DNA sequencing results confirmed that the recombinant lentiviral vectors pLKO.1-sh hDEK1 and pLKO.1-sh hDEK3 were successfully constructed. RT-PCR and Western blot results showed that the expression of DEK in human liver cancer cells BEL-7402 and Huh7 cells was higher, and pLKO.1-sh hDEK3 was more effective in inhibiting the DEK gene expression ( P < 0.05). Therefore, pLKO.1-sh hDEK3 was selected to infect BEL-7402 and Huh7 cells for subsequent functional experiments. CCK8 cell proliferation test result showed that the cell proliferation ability of BEL-7402 and Huh7 cells infected with recombinant lentivirus was weakened when compared with blank control and negative control group ( P < 0.05). Apoptosis results showed that the apoptosis rate of knockdown group was higher than that of blank and the negative control group ( P < 0.05). Cell scratch test result showed that the wound healing rate of knockdown group was lower than that of blank control and negative control group ( P < 0.05), and the difference was statistically significant; however, there was no statistically significant difference between blank control and negative control group. Conclusion:Targeting DEK expression in silent liver cancer cells can inhibit the cell proliferation, migration ability, and induce apoptosis, which lays the foundation for further study of the role of DEK gene in liver cancer.

Activation of inflammatory responses regulates the transmission of pain pathways through an integrated network in the peripheral and central nervous systems. The immunopotentiator thymosin alpha-1 (Tα1) has recently been reported to have anti-inflammatory and neuroprotective functions in rodents. However, how Tα1 affects inflammatory pain remains unclear. In the present study, intraperitoneal injection of Tα1 attenuated complete Freund's adjuvant (CFA)-induced pain hypersensitivity, and decreased the up-regulation of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6) in inflamed skin and the spinal cord. We found that CFA-induced peripheral inflammation evoked strong microglial activation, but the effect was reversed by Tα1. Notably, Tα1 reversed the CFA-induced up-regulation of vesicular glutamate transporter (VGLUT) and down-regulated the vesicular γ-aminobutyric acid transporter (VGAT) in the spinal cord. Taken together, these results suggest that Tα1 plays a therapeutic role in inflammatory pain and in the modulation of microglia-induced pro-inflammatory cytokine production in addition to mediation of VGLUT and VGAT expression in the spinal cord.

Objective The adverse drug reaction (ADR) reports of compound Kushen injection in Qingdao Central Hospital were analyzed to improve the level of safe and rational use of drugs.Methods We investigated 35 cases of severe ADR reports induced by compound Kushen injection in Qingdao Central Hospital from 2016.09 to 2018.03 retrospectively.Then the type of report,relevance evaluation and prognosis,ADR occurrence and recovered time,involved system/organs and main clinical manifestations were analyzed.Results The new moderate and new severe ADR incident were 51.4％ of 35 ADR reports.Twenty-nine cases of the ADR were occurred 30 minutes after injection.Twenty-three cases of ADR clinical symptoms can be recovered after 30 minutes.The main ADR was the digestive system reaction,followed by the neural system,respiratory system and cardiovascular system.Conclusions It is important to pay attention to the occurrence of compound Kushen injection ADR and strengthen its monitoring.

Objective To observe the effects of road transport on hematological and biochemical parameters in New Zealand rabbits.Methods A total of 12 healthy New Zealand rabbits were selected for 2 h road transport.Blood samples were collected at 0, 24, 48, 72 and 96 h after transport, respectively.White blood cells (WBC), red blood cells (RBC), hemoglobin (HGB), hematocrit (MCV), mean erythrocyte hemoglobin content (MCH), mean erythrocyte hemoglobin concentration (MCHC) and platelets (PLT) were measured using a blood analyzer.Blood alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (ALB), total protein (TP), urea nitrogen (UREA), creatinine, uric acid (UA), triglycerides (TG), total cholesterol (COHL), glucose (GLU), hypersensitive C-reactive protein (CRP), α-amylase (AMYL), and creatine kinase (CK) were detected by an automatic biochemical analyzer.Results Compared the parameters before and after transport, The WBC count was increased first (P< 0.05 or P < 0.01) and then decreased after transport, the levels of RBC, HGB, HCT and PLT were decreased first (P< 0.05 or P < 0.01) and then increased after transport, and MCV was significantly high at 96 h after transport (P< 0.05).Among the clinical biochemical parameters, ALT, AST and BUN were firstly elevated (P< 0.05 or P < 0.01) and then decreased.TP, ALB as well as CREA and TG were firstly decreased (P< 0.05 or P < 0.01) and then increased.GLU was significantly low at 24 h after transport (P< 0.05).All parameters except MCV at 96 h after transport were not significantly different from those before transport.Conclusions Changes of blood routine, liver and kidney function indexes, lipid metabolism indexes, glucose metabolism index and creatine kinase index are observed in the New Zealand rabbits after 2-hour road transportation, and all the indicators except MCV return to pre-transport levels within 96 h.

Objective To investigate the application of left transthoracic small incisions in combination with tubular gastrectomy for radical esophageal cancer surgery in elderly patients.Methods Clinical data of 83 patients with carcinoma in the middle or lower third of the esophagus aged ≥ 70 years who had undergone radical surgery at our hospital from January 2012 to September 2014 were collected.Patients were divided into two groups:the tubular gastrectomy group (n=38) which had undergone radical surgery through left transthoracic small incisions in combination with tubular gastrectomy and the stomach group (n =45) which had been treated with esophagectomy through conventional left posterolateral incisions and esophageal reconstruction with the remnant stomach.Clinical outcomes were compared between the two groups.Results Compared with the stomach group,operative blood loss [(204.3±75.4) ml vs.(258.4±80.2) ml,t=2.720,P＜0.05],chest drainage on the first and second postoperative day [(201.7±82.6) ml vs.(320.5±78.1) ml,(150.8±83.0) ml vs.(244.6 ± 81.2) ml,t =3.221 and 3.189,respectively,each P＜0.05],pain scores on the third postoperative day [(3.73 ± 1.02) vs.(0.24 ± 1.15),t =2.858,P ＜ 0.05],incidence of arrhythmia (13.2％ vs.33.3％,x2 =4.585,P＜0.05),pulmonary complications (18.4％ vs.46.7％,x2=7.353,P＜0.05) and incidences of gastroesophageal reflux and gastric emptying disorders (23.7％ vs.55.6％,0.0％ vs 11.1％,x2 =8.654 and 4.493,both P＜0.05) were less or lower in the tubular gastrectomy group.The two groups had no significant difference in operation time,length of hospital stay,anastomotic leakage,anastomotic stenosis or 1-year survival rate (each P ＞0.05).Conclusions Radical surgery with small incisions through the left chest combined with tubular gastrectomy shows highly favorable clinical outcomes in elderly patients with middle or lower esophageal cancer,and can significantly reduce surgical trauma,decrease postoperative complications,improve the quality of life,and help patients recover after surgery.

Objective To evaluate the effect of perioperative administration of Ambroxol combined with Ipratropium on elderly lung cancer patients undergoing thoracoscopic surgery.Methods 82 lung cancer patients aged ≥70 years who were scheduled for thoracoscopic lung resection were randomly assigned into 2 groups:the observation group(n=42) and the control group(n=40).Patients in the observation group were treated with 90 mg Ambroxol(mucosolvan) by intravenous drip and Ipratropium(atrovent) by atomizing inhalation,while patients in the control group were treated with an equal volume of 0.9％ sodium chloride solution.Pulmonary function and changes of arterial blood gases at admission,before surgery and 5 days after surgery,incidences of postoperative atelectasis and pulmonary infection,the rate of return to the intensive care unit(ICU),and the length of postoperative hospital stay were compared between the 2 groups.Results Compared with the control group,the percent predicted forced expiratory volume in 1 second (FEV1％),the ratio of forced expiratory volume in 1 second to forced vital capacity(FEV1/FVC％),the percent predicted maximum ventilation volume per minute (MVV％),and the arterial oxygen pressure (PaO2) were increased in the observation group before surgery [(87.0±8.2)％ vs.(80.6±7.6) ％,(90.4±6.4)％ vs.(81.4±7.0)％,(87.1±5.6)％ vs.(74.6±6.9) ％,(86.6±6.2) mmHg vs.(81.7±6.9)mmHg,t=2.477,2.588,2.937,3.405,respectively,allP＜0.05].TheFEV1％,FEV1/FVC％,MVV％ and PaO2 were higher in the observation group than in the control group 5 days after treatment [(76.4±9.2) ％ vs.(67.3±10.2) ％,(74.7±9.1) ％ vs.(63.0±11.2) ％,(69.5±9.2)％ vs.(60.1±9.2) ％,(79.5±11.5) mmHg vs.(70.1±11.8) mmHg,t 2.583,2.987,2.778,2.666,respectively,all P＜0.05].The incidences of postoperative atelectasis,pulmonary infection and the length of postoperative hospital stay were lower or less in the observation group than in the controlgroup[7.1％ vs.25.0％,11.90％ vs.32.50％,(8.5±1.8) days vs.(12.1±2.6) days,x2=4.897,5.072,2.351,respectively,all P＜0.05].No significant difference in the rate of return to ICU was found between the two groups(P＞0.05).Conclusions The combination of perioperative Ambroxol and Ipratropium can effectively improve lung function by improving ventilation and gas exchange function,reduce postoperative pulmonary complications and shorten the length of postoperative hospital stay in lung cancer patients aged 70 years and over.

Objective To clone and express a high mobility group box 1(HMGB1)protein of Schistosoma japonicum(Main-land strain)and analyze its function. Methods The DNA fragment of open reading frame encoding Sj HMGB1 protein was ampli-fied by RT-PCR from the mRNA of S. japonicum worms,then it was subcloned into the expression vector pET28a(+)to form the recombinant expression plasmid SjHMGB1-pET28a. The recombinant expression plasmid was transformed into the component E. coli BL21(DE3),and the tranformant containing recombinant expression plasmid was induced with IPTG to express the recombi-nant protein SjHMGB1. The recombinant SjHMGB1 protein was purified by affinity chromatography with nickel chelating affinity chromatography agarose gel. The Gel retard experiment and animal immunization were performed to analyze the DNA binding ca- pacity and the immunologic property of recombinant SjHMGB1. The expression levels of HMGB1 in different life cycle stages of S. japonicum were analyzed by Western bloting and RT-PCR. Female ICR mice were immunized with the recombinant SjHMGB1 pro-tein and infected with 45±2 cercariae of S. japonicum after three immunizations. Forty-two days post-infection,the worms and eggs of S. japonicum were recovered from the portal vein and liver tissue,respectively. The worm and egg reduction rates were calculat-ed respectively. Results A 530 bp of specific DNA fragment was amplified from mRNA of S. japonicum by RT-PCR,which was the open reading frame(ORF)encoding SjHMGB1protein confirmed by DNA sequencing analysis. The recombinant expression plasmid SjHMGB1-pET28a was constructed by cloning the ORF of SjHMGB1 into a expression vector pET28a(+). The bacterium transformants containing the recombinant plasmid expressed a soluble recombinant protein about 28 kDa after induced by IPTG, and the recombinant SjHMGB1 protein was purified by nickel chelating affinity chromatography. The gel retard experiment showed that the recombinant SjHMGB1 protein could bind to both supercoiled DNA and linear DNA,and the recombinant protein immu-nized mice produced high titers of antiserum IgG. Western bloting indicated that the recombinant SjHMGB1 protein was recognized specifically by the S. japonicum-infected mice serum. Above results showed that the recombinant SjHMGB1 protein possessed both functional activity and immunogenicity as the natural protein. RT-PCR and Western blot results showed that SjHMGB1 was abun-dantly expressed in the adult and egg stages whereas barely detectable in the cercaria stage. The immune protection experiment showed that the recombinant SjHMGB1 induced mice to produce high titers of specific antibody IgG but failed to conduct an effec-tive immune protection against S. japonicum. Conclusion The gene encoding HMGB1 from S. japonicum and the soluble recombi-nant SjHMGB1 protein with natural functional activity are obtained,and the recombinant SjHMGB1 has a high immunogenicity but is not able to induce an effective immune protection against S. japonicum.

Enolase is one kind of important glycolytic enzymes which widely exists in most organisms. A number of recent studies confirm that this enzyme has the functions of activating the plasminogen involving in the processes of infection and mi-gration of parasites reducing the immune function of the host as well as preventing parasites from the immune attack of the host. This paper reviews the current research advances in the parasite enolase and explores its potential for diagnosis drug develop-ment and vaccine target of parasitic diseases.

Objective To explore the association between disrupted in schizophrenia 1(DISC1) genepolymorphism and schizophrenia and different subtype depression.To verify if DISC1 gene is the common predisposing gene for schizophrenia and depression.Methods The genotypes and alleles in 260 cases of schizophrenicpatients,96 cases of depressive patients with psychotic symptoms,124 cases of depressive patients without psychotic symptoms,and 100 normal controls were examined with polymerase chain reaction(PCR),denaturing polyacrylamide gel elcctmphoresis separation technique.The association was analyzed between DISC1 gene single nncleotide polymorphisms(SNP) locus rs821616 and schizophrenia and different subtype depression.Results The frequeneies of the genotypes T/T,A/T,and A/A were 3.5%,28.0%and 69.5%respectively,the frequencies of alleles T and A were 9.6%and 90.4% respectively in schizophrenia group.The frequencies of the genotypes T/T,A/T,and A/A were 3.1%,24.0% and 72.9% respectively,the frequencies of alleles T and A were 15.6% and84.4% respectively in depression 1 group;The frequencies of the genotypes T/T,A/T,and A/A were 2.4%,23.4% and 74.2% respectively,the frequencies of alleles T and A were 15.3% and 84.7% respectively in depression 2 group;The frequencies ofthe genotypes T/T,A/T,and A/A were 1.0%,16.0% and 83.0% respectively,the frequencies of alleles T and A were 17.0% and 83.0% respectively in control group.There were significant differences in the frequencies of the genotypes (Chi-Square=8.072,P=0.045)and alleles(Chi-Square=8.564,P=0.036) of DISC1 gene among the four groups with non-parametric Kruskal-Wallis test.After pairwisecomparison each other in the four groups we found that there were significant differences in the frequencies of thegenotypes(Z=-2.802,P=0.005)and alleles(Z=-2.837,P=0.005) of DISC1 gene between patients withschizophrenia and normal controls with non-parametric Mann-Whitney test(two-tailed),there were no significantdifferences between other groups(P>0.05).Conclusion Our results suggest that DISC1 gene polymorphism is associated with schizophrenia significantly,but it is not associated with different subtype depression.This finding do not support the viewpoint that DISC1 gene is the common predisposing gene for schizophrenia and depression.