Growth inhibition induced by short hairpin RNA to silence survivin gene in human pancreatic cancer cells

BACKGROUND: Survivin is known to be overexpressed in various human malignancies, including pancreatic cancer, and mediates cancer cell proliferation and tumor growth, so the regulation of this molecule could be a new strategy for treating pancreatic cancer. In this study, short hairpin RNAs (shRNAs) specific to survivin were introduced into human pancreatic cancer Patu8988 cells to investigate the inhibitory effects on survivin expression and cell proliferation in vitro and in vivo. METHODS: Three kinds of shRNA specific to the survivin gene were designed and cloned into eukaryotic expression plasmid pGenesil-1 vector. Subsequently the recombinant plasmids were transfected into human pancreatic cancer Patu8988 cells with lipfectamine (TM) 2000 reagent. The mRNA and protein expressions of survivin in the transiently transfected Patu8988 cells were determined by RT-PCR, flow cytometry, and Western blotting analysis. The proliferation inhibition rates of stably transfected Patu8988 cells were determined by MTT assay. The antitumor activities of the three kinds of survivin-shRNA plasmids were evaluated in BALB/c nude mice inoculated with Patu8988 cells and bearing human pancreatic cancer. RESULTS: The three survivin-shRNA plasmids named pGenesil-1-survivin-1, pGenesil-1-survivin-2 and pGenesil-1-survivin-1+2 (with double interfering RNA sites) were successfully constructed, and were confirmed by restriction enzyme cutting and sequencing. At 48 hours after transfection, the expression of survivin mRNA and protein was inhibited in Patu8988 cells transfected with pGenesil-1-survivin-1, pGenesil-1-survivin-2, and pGenesil-1-survivin-1+2 when compared with that of either pGenesil-1-NC (with scrambled small interfering RNA) transfected cells or control cells (P<0.05). The MTT results showed that the proliferation rates of Patu8988 cells stably transfected with survivin-shRNA plasmids were reduced when compared with that of either pGenesil-1-NC transfected cells or control cells (P<0.01). Furthermore, when Patu8988 cells stably transfected with survivin-shRNA were injected into BALB/c nude mice, tumor growth was dramatically lower and the tumor was smaller than that of either pGenesil-1-NC transfected cells or control cells (P<0.01). The inhibitory effect of pGenesil-1-survivin-1 was the best among the three kinds of survivin-shRNA plasmids, but no combination of inhibitory effects was found in pGenesil-1-survivin-1+2. CONCLUSIONS: shRNAs specific to survivin have gene silencing effects and inhibit pancreatic cancer cell proliferation. shRNA activity against survivin could be of potential value in gene therapy for pancreatic cancer. However, shRNAs with double combining sites did not significantly enhance the interference compared with single site shRNAs, therefore further studies on this are needed.

Reevaluation of the effect of lamivudine therapy preoperative to prevent HBV recurrence after liver transplantation

BACKGROUND: Hepatitis B virus (HBV) recurrence may result in hepatic, insufficiency or dysfunction of liver grafts. This study was to reevaluate the preventive effect of lamivudine therapy pretransplant on HBV recurrence after liver transplantation with combined lamivudine and hepatitis B immunoglobulin (HBIG) as a prophylactic regimen. METHODS: This is a single-center, retrospective study of 122 liver transplant recipients operated on from January 2002 to September 2006 at our hospital. All subjects showed positive hepatitis B surface antigen (HBsAg) and HBV DNA in blood, without HEX mutation in YMDD at the time of liver transplantation. The protocol with combined larnivudine and HBIG for preventing HBV recurrence was used on the day of operation. The initial immunosuppression therapy was identical. After one year follow-up, the recipients were divided into 2 groups: patients without HBV recurrence (group]) and patients with HBV recurrence (group 11). Preoperafive larnivudine therapy and postoperative mycophenolate mofetil (MMF) and glucocorticoid therapy were analyzed using the Wilcoxon's test and Stepwise logistic regression method. RESULTS: In the HBV recurrence group, the duration of pre-transplant lamivudine administration was significantly longer than that in the without HBV recurrence group (Z=-4.424, P=0.000). The HBV recurrence rate was significantly higher in patients with preoperative lamivudine therapy than in patients without lamivudine therapy (chi(2)= 13.11, P=0.000); the risk of HBV recurrence increased by a 10.909-fold in patients with pre-transplant lamivudine therapy compared with that in patients without larnivudine therapy (OR=10.909; 95% Cl for OR: 2.86-41.67). Seven (63.6%) of 11 HBV recurrence recipients had YMIDD mutants. The duration of MMF or glucocorticoid was not different between the 2 groups (Z(MMF)=-1.453, P-MMF=0.146; Z(Prc)=-0.795, P-Prc=0.427). No significant difference was noted in the HBV recurrent rate in patients with MMF duration <= 6 and > 6 months ( Z 2= 0.185, P=0.667), as it was in patients with prednisone therapy <= 3 and > 3 months (chi(2) = 0.067, P= 0.793). CONCLUSIONS: With the protocol of combined lamivudine and HBIG for preventing HBV recurrence in liver transplantation recipients, liver transplantation candidates with positive HBV DNA should not be subjected to preoperative administration of larnivudine. A high dose of HBIG during the ahepatic period and in the early stage of post-transplantation can fulfill the treatment target as a long-term lamivudine therapy before liver transplantation. Long-term preoperative lamivudine treatment may result in an earlier HBV mutation in YMDD and increase the HBV recurrence rate and risk in the first year after transplantation.

Skeletal muscle cell apoptosis following motor nerve injury versus sensory nerve injury

Skeletal muscle atrophy inevitably occurs in denervated skeletal muscle, and cell apoptosis plays an important role in skeletal muscle atrophy and degeneration. The present study established rat models of simple nerve injury by transecting the ventral or dorsal spinal nerve root and observed rat skeletal muscle cell apoptosis following simple motor nerve injury versus simple sensory nerve injury. Following skeletal muscle denervation for 10 weeks, cell apoptosis was detected in skeletal muscle, which was accompanied by obvious changes in rat behavior and electrophysiological responses. In addition, changes in cross-sectional area and average gray-scale of motor endplates of the gastrocnemius muscle were analyzed following sciatic nerve injury and motor nerve injury. Cell nuclei in denervated skeletal muscle tissue were more densely arranged than in normal skeletal muscle tissue. Cell nuclei were most dense in the sciatic nerve injury group, followed by the motor nerve injury group and the sensory nerve injury group. Fas/FasL expression and the number of apoptotic cells increased in denervated skeletal muscle, and apoptosis-related changes were observed. These findings suggested that motor and sensory nerves provided trophic actions following skeletal muscle and motor nerve injury, resulting in a greater influence on skeletal muscle atrophy than sensory nerve injury. Therefore, reconstruction of motor nerves should be preferentially considered for treating denervation-induced skeletal muscle atrophy.

Light transmittance aggregometry with non-adjusted PRP is more suitable for clinical laboratory monitoring antiplatelet therapy

Objective In this study,maximum platelet aggregation rate of Light transmittance aggregometry (LTA) for coronary heart disease (CHD) patients taking antiplatelet drug and patients without antiplatelet therapy was measured in non-adjusted and platelet count-adjusted platelet-

Application of serum creatinine to prealbumin ratio in chronic heart failure patients with renal failure

Objective To investigate the application of serum creatinine to prealbumin ratio (Scr/PA) in the diagnosis of patients with heart failure complicated with renal failure.Methods This was a case-control study.Patients with chronic heart failure and heart failure complicated with renal failure admitted to Dalian Central Hospital from January 5,2020 to April 23.

Construction of m6A modification related prognostic model in older patients with FLT3 mutated acute myeloid leukemia

Objective To screen m6A modification-related genes,and to establish a prognostic model in patients with FLT3 mutated acute myeloid leukemia (AML),especially in older patients and to evaluate the prognostic efficiency of the model.Methods Gene expression omnibus (GEO) datasets were used to analyze abnormally expressed m6A enzymes and reading proteins in FLT3mutated AML;correlation analysis was used to screen m6A modified-related genes in expression profiles.

Clinical and microbiological features of community-acquired and nosocomial bloodstream infections in the surgical department of a tertiary-care hospital in Beijing

Background Bloodstream infections （BSls） remain a major cause of morbidity and mortality in patients undergoing surgery. This study aimed at elucidating the clinical characteristics of community-acquired BSIs （CABs） and nosocomial BSIs （nBSIs） in patients admitted to the surgical wards of a teaching hospital in Beijing, China. Methods This cross-sectional study compared 191 episodes of BSIs in 4074 patients admitted to the surgical wards between January 2008 and December 2011. Cases of BSIs were classified as CABs or nBSIs, and the characteristics, relevant treatments, and outcomes of CABs and nBSIs were compared. Results Of the 191 BSIs, 52 （27.2%） and 139 （72.8%）were CABs and nBSIs, respectively. Escherichia coli, coagulasenegative staphylococci, and Klebsiella spp, were the most frequently isolated microorganisms. There were significant differences between CABs and nBSIs with respect to the use of hormonal drugs, ventilation, acute physiology and chronic health evaluation （APACHE） Ⅱand American Society of Anesthesiologists scores, and prevalence of cancer （P 〈0.05）. Empirical antibacterial therapy did not decrease the crude mortality, but multivariate analysis showed that high APACHE Ⅱwas independently associated with a risk of mortality （odds ratio =0.97, 95% confidence interval： 0.93-1.02 for APACHE Ⅱ）. Conclusions We found significant differences in the clinical characteristics of surgical patients with CABs and nBSIs. The outcome of patients seems to be related to hiah APACHE Ⅱ scores.

Clinical variability in onset of influenza A （H7N9） infection

Clinical information of infections with the novel avian influenza A （H7N9） have been described elsewhere,but clinical variability, especially changes ot mental status in the onset of infection, has rarely been reported in patients infected with H7N9. In this report, the initial clinical manifestations of three confirmed cases are summarized. Two of the patients were in critical condition. In addition,

Tigecycline minimum inhibitory concentration for carbapenem-resistant Acinetobacter baumannii by two methods

To the editor： Carbapenem-resistant Acinetobacter baumannii （CRAB） is one of the most representative multidrug-resistant gram-negative bacteria, and its resistant rate has steadily increased in recent years.1.2 CRAB infections can currently be treated with tigecycline. However, with its rapid promotion and wide application in clinical therapy,

In-vitro activity of tigecycline against clinically common multiple-drug-resistant pathogens

Objective To investigate in-vitro activity of tigecycline against common clinical multidrug resistant(MDR) bacteria.Methods The MDR bacteria(except Pseudomonas aeruginosa) were collected retrospectively from 2009 to 2010 in 15 hospitals from 11 cities.The MIC of

Clinical application of automated digital image analysis for morphology review of peripheral blood leukocyte

Objective To explore the clinical application of automated digital image analysis in leukocyte morphology examination when review criteria of hematology analyzer are triggered.Methods The reference range of leukocyte differentiation by automated digital image analysis was established by analyzing 304 healthy blood samples from Peking University First Hospital.Six hundred and ninety-seven blood samples from Peking University First Hos-

Selection and evaluation of clinical diagnostic cut point of glycated albumin

Objective To investigate the relationship between GA and HbA1c,to identify the optimal GA cut-off values used to diagnose preDM and DM,and to verify the diagnostic significance of blood glucose biomarkers.Methods A total of 20 491 samples from Chinese people were involved in the correlation analysis.The participants were expected to complete the test of FPG,HbA1cand GA.

Clinical performance of serum proprostate-specific antigen isoform[-2] proPSA in prostate cancer

Objective To evaluate the application value of isoform[-2]proprostate-specific antigen(p2PSA),percentage of p2PSA(%p2PSA)and prostate health index(PHI)in cases with slightly elevated serum PSA.Methods A case-control study was adopted.The clinical data and sera of patients who had total PSA(t PSA)val-

The clinical application value of glycosylated hemoglobin(HbA_1c) plus fasting plasma glucose in diagnosing diabetes

Objective To evaluate clinical application value of HbA1c plus fasting plasma glucose(FPG) in diagnosing diabetes mellitus(DM).Methods 681 patients with DM were enrolled in this study. The level of HbA1c by Bio-RAD-10 glycated hemoglobin analyzer,FPG by

Pyrosequencing for detection of drug resistant relevant mutation in the polymerase gene of hepatitis B virus and its clinical application

Objective To explore the accuracy and clinical application of pyrosequencing for detection of drug resistant relevant mutation in the polymerase gene of hepatitis B virus(HBV).Methods Compared with Sanger sequencing,the accuracy and sensitivity of pyrosequencing were assessed.Pyrosequencing was used to determine the serum of 1 164 patients with chronic Hepatitis B and its re-sults were analyzed.Results The sensitivity of pyrosequencing was 1×103KIU/L,the same as Sanger sequencing.

Analysis of pathogen spectrum and antimi-crobial resistance of clinical common organisms from twelve teaching hospitals in 2016

Objective To investigate the spectrum and antimicrobial resistance of major pathogens causing nosocomial infections in China,2016.Methods Non-duplicated nosocomial cases as well as pathogens causing bloodstream infections(BSI),hospital-acquired pneumonia(HAP)and intra-abdominal infections(IAI)from 12 teaching hospitals across China were collected.The minimum inhibitory concentrations(MICs)of important clinical common strains were determined by agar dilution method or broth microdilution method.The CLSI M100-S27 criteria was used for interpretation.Data were analyzed by using WHONET-5.6 software.Results A total of 2 060 cases were collected,including 894 cases from BSI,630 cases from HAP and 536 cases from IAI.The MICs of 1 896 important clinical common strains were determined.Escherichia coli and Klebsiella pneumoniae were the most prevalent pathogens causing BSI and IAI,while Acinetobacter baumanii and Pseudomonas aeruginosa were dominated in HAP.All Staphylococcus aureus were susceptible to tigecycline,linezolid,daptomycin and glycopeptides.Methicillin-resistant S.aureus accounted for 44.4%(75/169)of all the S.aureus.The rate of methicillin-resistant coagulase-negative staphylococci was 80.9%(72/89).No Enterococcus strains were found resistant to tigecycline,linezolid or daptomycin.Vacomycin resistant enterococcus was found in Enterococcus faecium,accounting for 1.8%(2/11)of all E.faecium strains.Tigecycline,meropenem,amikacin,imipenem,and polymyxin B exhibited high potency against Enterobacteriaceae and the susceptibility rates were 96.6%(865/895),94.3%(859/911),94.2%(858/911),94.1%(857/911),and 91.6%(820/895),respectively.The prevalence of extended-spectrum β-lactamase was 58.4%(263/450)in E.coli and 28.6%(84/294)in K.pneumonia.The rate of carbapenem resistant K.pneumonia and E.coli was 15.3%(45/294)and 1.8%(8/450),respectively.The percentage of polymyxin B resistant K.pneumonia and E.coli was 4.1%(12/294)and 4.4%(20/450),respectively.The rate of tigecycline resistant K.pneumonia and E.coli was 2.4%(7/294)and 0.2%(1/450),respectively.A.baumanii showed low susceptibility to the antimicrobial agents except tigecycline(91.4%,235/257)and polymyxin B(100%,257/257).The rate of carbapenem resistant A.baumanii was 80.5%(207/257).The rate of carbapenem resistant P.aeruginosa was 31.7%(59/186).Polymyxin B and amikacin demonstrated high antibacterial activity against P.aerug-inosa with susceptility rate of 100%(186/186)and 90.9%(169/186),respectively.Conclusion Nosocomial pathogens showed high susceptibilities against tigecycline and polymyxin B.Antimicrobial resistance in A.baumannii is a serious problem.The prevalence of carbapenem-resistant Enterobacteriaceae and polymyxin B resistant Enterobacteriaceae has increased,which should be monitored continuously in China.

Clinical significance of urine ferritin in patients with cerebral infarction

Objective To find an indicator in urine to assist diagnosis of cerebral infarction,we investigated the changes of urine ferritin in patients with cerebral infarction.Methods Collected serum from 30 healthy volunteers and 53 patients with cerebral infarction (CI) ,with ratio ofmales to

The expression of Panton-Valentine leukocidin gene and its correlation with agrA among different clinical isolates of Staphylococcus aureus

Objective To investigate the correlation between expression of Panton-Valentine leukocidin gene and accessory gene regulator among different clinical isolates of Staphylococcus aureua.Methods All non-duplicate Staphylococcus