Objective:To construct and identify a plasmid vector of short interfering RNA(siRNA)on pneumocystis carinii(PC) major surface glycoprotein(MSG)gene's upstream conserved sequence(UCS).Methods:Short hairpin RNA(shRNA) oligonucleotide targeting PC MSG UCS gene which was chemically synthesized and inserted into RNAi-Ready plasmid vector pTZU6+1 after annealing.The recombinant plasmid,PC-UCS,transformed into E.coil.TOP10 and amplified,was digested by restriction endonucleases HindⅢand EcoRI and identified by gel electrophoresis and DNA sequencing.Results:Gel electrophoresis and DNA sequencing showed that the recombinant plasmid containing the correct and full shRNA oligonucleotide. Conclusion:The siRNA plasmid,pPC-UCS,was constructed successfully.

0.05). ⑤ berberien increased basal cGMP concentrations directly (P

The brief discussion is introduced in the paper on the academic thought of professor YAN Jie, the contemporary famous TCM doctor, on functional dyspepsia treated with acupuncture and moxibustion. Treatment based on "the three-regional acupoint selection" is applied to professor YAN's treatment for functional dyspepsia, in which, acupuncture is on Sibai (ST 2), Liangmen (ST 21) and Zusanli (ST 36), and the supplementary points are added accordingly. The academic thought is described as the combination of acupuncture and moxibustion based on strengthening healthy qi, supplemented by soothing the liver and psychological counseling. Also, an example is provided.
Acupuncture ; education ; history ; Acupuncture Points ; Acupuncture Therapy ; history ; Adult ; Dyspepsia ; history ; therapy ; Female ; History, 20th Century ; Humans ; Moxibustion

Objective To investigate the immunophenotype and the cytotoxicity of cytokine-induced killer(CIK) against tumor cells in vitro.Methods Lymphocytes cells were isolated freshly from peripheral blood of healthy donors by Ficoll-Hypaque density centrifugation,and the cells obstained were induced by IFN?,IL-2 and CD3McAb.Phenotypes and cytotoxicity of CIK were analysed by FACS.Target cells were differentiated from effect cells by CFSE dying.The mortality of Target cells were determined by FACS.Results The maximum proliferation of CIK reached at the 22nd day.The phenotypes of CD3,CD11a,CD54,HLA-DR were expressed highly;CD25,CD28,CD69,FasL were expressed moderately on CIK.The expression of CD16 was not increased.CIK possessed the cytotoxicity against tumor cells of K562,HL-60,Hela,SMMC7721 and A375.Conclusion IFN?,CD3McAb,IL-2 can induce peripheral lymphocytes to produce CIK which own strongly proliferation and exert highly efficient cytotoxic effects on tumor cells.

Dyslipidemia is a risk factor for stroke, and reducing low-density lipoprotein cholesterol (LDL-C) levels can decrease the risk of ischemic stroke. Both statins and PCSK9 inhibitors can effectively reduce LDL-C levels, but some patients cannot tolerate statins, and PCSK9 inhibitors are expensive. Animal experiments have shown that ezetimibe can reduce oxidative stress and endoplasmic reticulum stress response, inhibit cell apoptosis, increase autophagy, and protect nerve tissue by activating AMP activated protein kinase (AMPK) signaling pathway. Clinical studies have shown that ezetimibe can further reduce LDL-C levels on the basis of statin therapy, and reduce the risk of ischemic stroke in patients with acute coronary syndromes, and there is no obvious adverse reaction. However, ezetimibe cannot reduce the incidence of stroke in elderly people who are ≥75 years old, have elevated LDL-C and do not have coronary heart disease. At present, there are few studies on the use of ezetimibe for secondary prevention of ischemic stroke, and its effect needs to be further confirmed by large randomized controlled trials.

Foreign body management is an important aspect in the practice of a general practitioner1. In Singapore, the most common foreign body we encounter would be fishbone. This is not surprising as steamed fish is a common dish in our Asian diet. In spite of the rapid advances in medical technology, a good history taking and physical examination remain the most important step in helping us clinch a diagnosis. In this article, we will focus on the common foreign bodies seen in the Ear, Nose and Throat area as a general practitioner, and the tips and pearls in the examination and management. In each section, there will also be some tips on the red flags to look out for and when a tertiary referral should be made.

Objective To observe the protective effect of different doses sodium hydrosulfide (NaHS) as a donor of hydrogen sulfide on renal ischemia reperfusion injury (IRI).Methods Twenty-eight healthy male Wistar rats were randomly divided into 4 groups,namely sham operation (Sham) group (n =7),renal ischemia reperfusion (IR) group (n =7),sodium hydrosulfide (NaHS) high dose group (n=7),sodium hydrosulfide low dose group (n=7).After excision of the right kidney,two dose NaHS group (300 nmol/min,1.5 μmol/min) received 15-minute continuous administration via left renal artery.Sham group and IR group received same volume of saline.Five minutes after stopping drug,the left renal pedicle in NaHS group and IR group was clipped with no damage arteriole occlusion.After 45 minutes blockade,a model of acute renal ischemia reperfusion injury was established.Sham group underwent the same procedure as model groups without clamping the left renal artery.Specimens of renal tissue and blood were harvested at 24-hour after blood flow restore in the kidney.BUN and Scr were measured.Kidney pathological damage was semi-quantitatively analyzed.Production of H2S in renal tissue was detected.Expression of cystathionine γ lyase (CSE) and cystathionine β synthase (CBS) mRNA in kidney tissue was examined by real-time PCR.Results Compared with sham group,BUN and Scr increased significantly (P＜0.01),kidney tissue H2S production decreased significantly (P＜0.01),mRNA expression of CSE and CBS down-regulated significantly (P＜0.01) in IR group.The kidney pathology of sham group was normal,while acute tubular necrosis was found in IR group.Compared with IR group,BUN and Scr decreased significantly (P＜0.01),kidney tissue H2S production increased significantly (P＜0.05),mRNA expression of CSE and CBS up-regulated significantly (P＜0.01) in NaHS groups.Pathological damage of acute tubular necrosis was significantly improved in NaHS pretreatment group.There was no significant difference between two NaHS doses groups.Conclusion Hydrogen sulfide has a protective effect on renal IRI.

Objective:To investigate the antitumor efficiency of the special cytotoxic T lymphocytes(CTLs) activated by dendritic cells(DCs) pulsed with K-ras (12-Val) antigen.Methods:DCs was generated from PBMC in the presence of granuloceyte/macrophage-colony stimulating factor(GM-CSF),interleukin-4(IL-4)in vitro.DCs were differently sensitized with K-ras mutant pancreatic cancer cell line,K-ras(12-Val) mutant peptide,K-ras(12-Val) mutant peptide with the surface of cationic nanoparticle.Cell surface markers on DCs was measured by flow cytometry.The activation of CTL induced by DCs was detected by ~3H- thymidine incorporation test.The killing effects of CTL to pancreatic cancer was detected by ~(125)I-UdR release test. Production of IL-12 and IFN-γ by DCs and PBMC was detected by ELISA.Results:Compared with DCs pulsed with K-ras(12-Val) mutant peptide and K-ras (12-Val) mutant peptide with the surface of cationic nanoparticle,DCs pulsed with whole tumor antigen could better induce CTLs killing activity(P＜0.05).The DCs with K-ras(12-Val) mutant peptide and K-ras mutant peptide with the surface of cationic nanoparticle could produce specific CTL killing activity aganist pancreatic cancer cell line Patu8988(K-ras+)(P＜0.05),but not SW1990(K-ras-)(P＞0.05). K-ras (12-Val) mutant peptide with the surface of cationic nanoparticle at lower concentrations can be effectively presenting on the surface of DCs than only K-ras (12-Val) mutant peptide.Conclusion:K-ras (12-Val) mutant peptide with cationic carrier can be effectively presenting and expression of DCs and induce CTL specific killing activity aganist pancreatic cancer cell lines with K-ras (12-Val) mutant peptide.