Objective: To investigate whether the expression of Apollon (an apoptosis inhibitory gene) silenced by using RNA interference (RNAi) technology in combination with tetramethylpyrazine (TMP) intervention can increase the sensitivity of human myeloid leukemia K562 cells to vincristine (VCR) and daunorubicin (DNR). Methods: Using the best small interference RNA (siRNA) targeting Apollon gene which was screened out in a previous experiment, the eukaryotic expression vector pGPHI-GFP-Neo-Apollon was constructed. Then, the K562 cells were transfected with pGPHI-GFP-Neo-Apollon by LipofectamineTM 2000 and screened out by G418. After Apollon siRNA was transfected stably into K562 cells, the expressions of Apollon mRNA and protein were detected by reverse transcription (RT)-PCR and immunofluorescence, respectively. The sensitivities to chemotherapeutic agents VCR and DNR after RNAi in combination with TMP intervention were detected by MTT method, and apoptosis rate was detected by flow cytometry. Results: pGPHI-GFP-Neo-Apollon vector was constructed successfully and it could express stably in K562 cells, which silenced the expressions of Apollon mRNA and protein effectively. The sensitivity to VCR or DNR increased significantly after RNAi in combination with TMP intervention. The half inhibitory concentration (IC50) values of VCR and DNR were significantly lower than those in the group of TMP intervention alone (P < 0.05). The apoptosis rate of K562 cells after transfection with Apollon siRNA in combination with TMP intervention was significantly increased comparing with that of the untreated control group and TMP intervention alone group (P < 0.05). Conclusion: RNAi targeting Apollon gene in combination with TMP intervention can obviously increase the sensitivity to VCR or DNR in K562 cells and significantly reduce the dosage of chemotherapeutic agents as well as improve apoptosis induced by chemotherapy. These results suggest that Apollon gene silenced by RNAi in combination with TMP intervention may reverse the drug resistance of leukemia cells. Copyright © 2013 by TUMOR.

Objective There are two main functions of gastrointestinal tract,digestion and absorption,and barrier function.The latter has an important defensive effect,which keeps the body away from the invading and damaging of bacteria and endotoxin.It maintains the systemic homeostasis.Intestinal dysfunction would happen when body suffers from diseases or harmful stimulations.The more serious intestinal disorders would harm the intestinal protective mechanism,or intestinal barrier function,and bacterial/endotoxin translocation,of intestinal failure(IF) would ensue.This article provides a critical review of the evidence indicating that an increase in bacterial translocation is associated with sepsis,and even the multiple organ failure syndrome in critically ill patients.The in-transit microorganisms play an essential role in the homeostasis of local and systemic immunity.Methods All studies published from 2000 to June 2005 about intestinal permeability,bacterial translocation,and systemic inflammatory response syndrome were located by search of PubMed.Results Clinical and experimental studies investigating the correlation between bacterial translocation and systemic inflammatory response syndrome,associated with the damage of the gut barrier function.To keep the mucosal barrier function intact is one of the main issues in the prevention of bacterial translocation.This could be achieved by the adequate delivery of oxygen and nutrient supplementation to the gut.Enteral nutrition,probiotic can be a good choice.Conclusion With a better understanding of the bacteria-host interactions in health and the alterations induced by critical illness,new therapies that improve the environment of both may lead to better recovery rates in intensive care unit patients.

Objective To analyse the clinical and pathological characteristics of patients with myopathies that were misdiag-nosed as myasthenia gravis because of external ophthalmoplegia ,widen the thoughts for differential diagnosis of extraocular muscle paralysis .Methods The clinical and pathological features of 11 myopathy cases with ptosis and diplopia admitted to the neurology department of the First Affiliated Hospital of Chongqing Medical University between October 2010 and May 2014 were retrospec-tively reviewed and analyzed .Results Among the 11 patients ,6 male and 5 female ,aged 16-66 years old .All had paralysis of ex-traocular muscle manifestations ,including oculopharyngeal muscular dystrophy(OPMD) in 3 cases ,oculopharyngeal distal myopa-thy in 2 cases ,chronic progressive external ophthalmoplegia(CPEO) in 6 cases .Muscle biopsy showed the characteristic pathologi-cal changes .Statistical analysis showed that the age of onset in OPMD patients was older(P<0 .05) ,creatine kinase level in oculo-pharyngeal distal myopathy patients was higher ,but the difference was not significant (P>0 .05) .Conclusion Certain rare myopa-thy should be considered for patients with external ophthalmoplegia seemingly myasthenia gravis .Muscle biopsy can provide clue for differential diagnosis .

Objective To observe the clinical efficacy and safety of nicergoline and aniracetam in treatment of mild and moderate cognitive dysfunction after cerebral infarction .Methods Infarction were enrolled after 70 cases of mild and moderate cognitive im-pairment patients were randomly divided into nicergoline treatment group (observation group) and aniracetam treatment group(con-trol group) ,all 35 cases .After 12 weeks of treatment were compared before and after treatment mini mental state examination (MMSE) ,activities of daily living scale(ADL) scores and transcranial Doppler(TCD) to improve the situation .Results After 12 weeks ,both groups after treatment compared with treatment MMSE score was significantly higher (P< 0 .05) ,and nicergoline group than aniracetam group(P< 0 .01);ADL score before treatment were significantly reduced (P< 0 .05) ,TCD improvement nicergoline group than aniracetam group (P<0 .01) .Conclusion Nicergoline mild and moderate cognitive dysfunction after cerebral in-farction have significant effects ,and better than aniracetam ,no significant adverse reactions occur for a wide range of clinical applications .

Objective To study the clinical and pathologic features,histological criteria and pathologic factors contributing to diagnosis of mixed epithelial and mesenchymal tumors(mixed m?llerian tumors,MMT)of the uterus.Methods A retrospective study of 102 cases of MMT of the uterus (74 adenofibromas including 9 recurrent cases,3 atypical polypoid adenomyomas,2 carcinofibromas,10 adenosareomas and 13 carcinosarcomas)was undertaken.Clinical records,gross features and tissue slices were reviewed.The follow-up data were analysed.Results The most common symptom was vaginal bleeding.Clinical signs included pelvic mass,uterine polyps,and enlarged uterus.Benign MMT usually presented as exophytic polypoid masses extending into the uterine cavity or protruding through the external os,often broad-based,lobulated and papillary.It was hard to distinguish low-grade malignant MMT from the benign ones by gross appearance.High-grade malignant MMT had the common gross features of carcinoma and sarcoma.Histologically,MMT showed a biphasic differentiation of mesenchymal and epithelial components.MMT were classified according to whether these elements were benign or malignant.Nine cases of adenofibroma without unique features for the diagnosis of adenosarcoma recurred at postoperative intervals of 3 to 96 months.Recurrent tumors were almost always confined to the original site.Conclusions Uterine MMT tumors according to WHO diagnostic criteria are not rare.The differential diagnosis depends on a multifactorial analysis.The recurrent adenofibromas may be a kind of borderline tumors with benign appearances and malignant behavior.

To compare the activity of RGD-TRAIL in different expression systems, RGD-TRAIL in both Escherichia coli (E.coli) and Pichia pastoris was constructed and expressed. In vitro activity of RGD-TRAIL from Pichia pastoris expression system was also analyzed. Genetic engineering techniques were used to construct recombinant plasmid pET30-rgd-trail and pHBM-rgd-trail. The recombinant protein RGD-TRAIL was purified with Ni ion affinity chromatography after induction. MTT assay, ELISA, scratch wound healing, transwell migration assay and Hoechst 33342 staining were performed to detect the effects of RGD-TRAIL on proliferation, binding activity, migration and apoptosis. The expression of apoptosis-associated proteins was detected by Western blotting. Recombinant protein RGD-TRAIL was successfully expressed in a form of inclusion body in E.coli, while expressed secretorily in Pichia pastoris. It possessed more potent cytotoxicity than RGD-TRAIL in E.coli by MTT assay. The RGD-TRAIL expressed by Pichia pastoris showed powerful binding affinity with cancer cells expressing α(v), DR4, DR5 and highly potent cytotoxicity through inducing apoptosis of cancer cells. Nuclear fragmentation was examined by Hoechst 33342 staining. Cleaved PARP and caspase-3 were also detected after incubation with RGD-TRAIL. Additionally, RGD-TRAIL inhibited migration significantly in A549 and HT1080 cells. The results demonstrate that Pichia pastoris expression system is more suitable for the recombinant protein RGD-TRAIL. Its binding affinity and antitumor activity might make RGD-TRAIL a promising candidate for cancer therapy.
Antineoplastic Agents ; pharmacology ; Apoptosis ; Blotting, Western ; Cell Line, Tumor ; Chromatography, Affinity ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; Humans ; Oligopeptides ; biosynthesis ; pharmacology ; Pichia ; metabolism ; Plasmids ; Recombinant Fusion Proteins ; biosynthesis ; pharmacology ; TNF-Related Apoptosis-Inducing Ligand ; biosynthesis ; pharmacology

Electric Stimulation ; Humans ; Hypoglossal Nerve ; Sleep Apnea, Obstructive ; therapy

DNA, Viral ; genetics ; Hep G2 Cells ; Hepatitis B virus ; drug effects ; Humans ; Oligonucleotides, Antisense ; pharmacology

Biobank is a biorepository which organized for collecting and storing human biospecimen as well as associated information for research uses.Biobank is the fundamental platform which translates the basic research result into clinical practice.It also plays an important role in disease diagnosis,new drug development,disease-related genetic research and epidemiological studies.The rise of translational medicine promotes the construction and development of the biobank.This review highlights the necessity to establish biobank,and focuses on the recent advances of the modem type of biobank,quality control of biospecimens,construction of biobank,best practices and guideline applied for biobank.This review also provides information for improvement of biobank.

Objective To analyze the predictive value of N-terminal B-type natriuretic peptide precursor(NT-proBNP) on auricular thrombosis in patients of coronary heart disease (CHD) combined with atrial fibrillation (AF).Methods The clinical data of 124 patients of CHD combined with AF were analyzed retrospectively.All patients were divided into research group (60 cases,compliance with auricular thrombosis) and control group (64 cases,non-compliance with auricular thrombosis),according to the occurrence of auricular thrombosis.The values of NT-proBNP,fasting plasma glucose (FPG),postprandial plasma glucose(2 h PG),total cholesterol (TC),high density lipoprotein cholesterol (HDL-C),oxidized low density lipoprotein (ox-LDL),high sensitivity C reactive protein (hs-CRP) and left ventricular ejection fraction (LVEF),left ventricular end diastolic diameter (LVEDd) and left atrial diameter (LAD) were detected and compared between two groups.The relative factors to the occurrence of auricular thrombosis were confirmed by multivariate Logistic analysis.The best cutoff point of NT-proBNP was confirmed by the areas under the receiver operating characteristic curve (ROC).Results The values of NT-proBNP,ox-LDL,hs-CRP,LVEDd and LAD in research group were higher than those in control group [(4 312.6 ± 209.1) pmol/L vs.(3 421.6 ± 156.8) pmol/L,(4.0 ± 0.9) mmol/L vs.(3.4 ± 0.8) mmol/L,(7.4 ± 1.3)mg/L vs.(5.8 ± 1.0) mg/L,(74.3 ± 6.8) mm vs.(58.1 ± 5.5) mm,(39.6 ± 4.3) mm vs.(32.5 ± 3.8) mm],LVEF and HDL-C were lower than those in control group [(48.2 ± 3.1)％ vs.(57.3 ± 3.8)％,(0.72 ± 0.16)mmol/L vs.(1.08 ±0.27) mmol/L],and there were significant differences (P ＜0.05).There were no significant differences in the values of FPG,2 h PG and TC between two groups (P ＞ 0.05).Multivariate Logistic analysis showed that NT-proBNP and LAD were independent factors to the occurrence of auricular thrombosis (P =0.009,0.028).There was significant difference in the occurrence of auricular thrombosis between patients with NT-proBNP ＞ 4 250 pmol/L and patients with NT-proBNP≤4 250 pmol/L (P =0.028).Conclusion NT-proBNP is an independent predictor for the occurrence of auricular thrombosis to patients of CHD combined with AF.